Telomerase activity and cytokeratin 20 as markers for the detection and followup of transitional cell carcinoma: an unfulfilled promise.

PURPOSE: Telomerase activity compensates for the erosion of chromosomes and it has been detected in a wide variety of human tumors. Cytokeratin 20, an intermediate filament of epithelial cells, is expressed particularly in the urinary tract. These 2 molecules are candidates to become markers for the detection and followup of bladder carcinoma. We evaluate whether each molecule may serve as a potential marker and whether the 2 combined would improve the detection or followup of bladder carcinoma in a noninvasive manner. MATERIALS AND METHODS: We obtained 44 morning urine samples from patients with transitional cell carcinoma patients and 26 from age matched patients with a wide variety of clinical disorders but no malignancy of any kind. A telomerase polymerase chain reaction-enzyme-linked immunosorbent assay kit was used to determine telomerase activity and cytokeratin 20 expression was determined by nested reverse transcriptase-polymerase chain reaction. RESULTS: All samples tested positive for cytokeratin 8 expression, which verified epithelial cells in the urine samples. Of the 44 transitional cell carcinoma cases of all stages and grades 37 (84.1%) were positive for telomerase activity, 36 (81.8%) were positive for cytokeratin 20 expression and 65.9% were double positive. Of the 29 controls with various clinical conditions other that malignancy 22 (75.9%) were positive for telomerase activity, 13 (44.83%) were positive for cytokeratin 20 expression and 34.6% were double positive. CONCLUSIONS: Telomerase activity and cytokeratin 20 expression are not specific for malignancy and may be detected in many nonmalignant pathological conditions. Therefore, their use as potential markers of bladder carcinoma should be carefully reevaluated.

Kidney function after radical nephrectomy: assessment by quantitative SPECT of 99mTc-DMSA uptake by the kidneys.

UNLABELLED: To determine the function of the remaining contralateral kidney after the removal of a functioning kidney, 30 consecutive patients (18 men, 12 women; average age, 67 y; age range, 34-87 y) who were undergoing unilateral radical nephrectomy were evaluated by sequential quantitative 99mTc-dimercaptosuccinic acid (DMSA) SPECT (QDMSA) studies. METHODS: The 30 patients were undergoing radical nephrectomy for renal tumors. The first study was done before surgery. Follow-up studies were performed 2-23 mo after surgery. Clinical evaluations and determinations of serum creatinine level were performed at the same time as the QDMSA studies. RESULTS: The relative contribution of the resected kidneys to the global renal function before surgery was 43.2% +/- 7.3%. After surgery the uptake of the remaining kidney increased from 13.4% +/- 4.0% to 18.3% +/- 5.8% (t = 5.7; P = 0.0000). The relative function of the remaining kidney increased from 56.8% +/- 7.1% to 79.1% +/- 23.6% (t = 4.9; P < 0.0001) of the global renal function before nephrectomy. Increases in the renal volume (from 211 +/- 62 cm3 to 229 +/- 68 cm3; t = 4.5; P = 0.0001) and in the percentage injected dose per cubic centimeter (%ID/cm3) of the remaining kidney (from 0.066 +/- 0.02 % ID/cm3 to 0.085 +/- 0.03 %ID/cm3; t = 4.6; P = 0.0001) were associated with this change. Nine patients had 2 follow-up studies performed 3-4 mo after surgery and 12-14 mo after surgery. The volume of the remaining kidney (209.22 +/- 46.20 cm3 versus 217.88 +/- 58.85 cm3; t = 0.962; P = 0.364), the %ID/cm3 (0.09 +/- 0.016 %ID/cm3 versus 0.093 +/- 0.025 %ID/cm3; t = 0.362; P = 0.726), and the percentage uptake (19.26% +/- 4.45% versus 20.11% +/- 7.01%) did not change significantly between these 2 QDMSA studies. CONCLUSION: The results of this study suggest that adaptive changes causing hyperfunction of the remaining kidney may occur after nephrectomy of a functioning kidney in adults. These changes occur soon after surgery, persist for at least 1 y, and are evident on QDMSA studies.

Urinary cytokeratin 20 as a marker for transitional cell carcinoma.

OBJECTIVES: To determine if detection of cytokeratin 20 (CK20) gene expression, by reverse transcriptase-polymerase chain reaction (RT-PCR) in urine from transitional cell carcinoma (TCC) patients, can provide a new noninvasive tool for the follow-up of patients with urothelial carcinoma of the bladder. METHODS: Urine was collected from 95 patients previously diagnosed as TCC during their follow-up, and from 27 healthy volunteers. All patients had a transurethal resection of tumor or biopsies obtained from 'suspicious' areas in the bladder. RNA was extracted from cells collected from the urine and RT-PCR was performed with specific primers for the amplification of cytokeratin 8, a general marker for epithelial cells, and of CK 20, a marker for TCC urothelium. RESULTS: CK20 expression was detected in 86.7% of TCC patients, and only in 3.3% of healthy volunteers (specificity 96.7%). Strong correlation was found between tumor grade and expression of CK20 in urine. All grade III and IV tumors demonstrated positive CK20 expression (100% sensitivity), whereas the sensitivity for lower grades was between 71 and 80%. Among 11 patients with a previous biopsy-proven diagnosis of TCC and a current negative biopsy, in 9 patients CK20 expression was detected. Further follow-up of these patients for a period of 6 months revealed recurrence of TCC in 4 patients. CONCLUSION: CK20 detection in urine cells is a simple, noninvasive method with a high potential to become the marker of choice for monitoring and follow-up of TCC patients. More information is needed regarding CK20 expression in nonmalignant urological disease, to evaluate its use for routine screening purposes.

The additional value of free prostate specific antigen to the battery of age-dependent prostate-specific antigen, prostate-specific antigen density and velocity.

This study describes the value of using the fraction of free prostate-specific antigen as a further marker in the early detection of prostate cancer. This newly introduced marker is compared to the usual battery of age-dependent total prostate-specific antigen, prostate-specific antigen density (microg/l x g tissue) and prostate-specific antigen velocity (microg/l x year). Determination of total prostate-specific antigen and free prostate-specific antigen was performed on fresh serum samples obtained from 3470 symptomatic patients aged 45-80 attending the Urology Clinics, or their General Practitioners. Among them, 310 patients had total prostate-specific antigen above the age-dependent cut-off, and/or free/total prostate-specific antigen under 11%, with different prostate-specific antigen densities and velocities. Only 147 patients complied to undergo biopsy: in 72 of those patients, benign prostatic disease was histologically confirmed, while in 75 patients primary prostate cancer was histologically confirmed. Total and free prostate-specific antigen levels were determined using the third generation DPCs prostate-specific antigen assay performed on the Immulite automated immunoassay instrument. Total prostate-specific antigen age reference values were adopted from Oesterling et al. (J Am Med Ass 1993; 270:860-4); the prostate-specific antigen density was considered suspicious of prostate cancer if it was greater than 0.15 microg/l prostate-specific antigen per gram tissue (Seaman et al. Urol Clin N Am 1993; 20:653); prostate-specific antigen velocity greater than 0.75 microg/l x year (Carter et al., J Am Med Ass 1992; 267:215) was considered suspicious for prostate cancer. Of the 147 patients, 75 had prostate cancer and 72 had benign prostatic hypertrophy. The difference between prostate cancer and benign prostatic hypertrophy was significantly reflected only by free/total prostate-specific antigen and prostate-specific antigen velocity. These parameters also provided the best sensitivity and specificity. Only these parameters proved to be significant when using a backwards logistic regression model (prostate-specific antigen velocity, p = 0.007 odds ratio 2.782; free/total prostate-specific antigen %, p = 0.016 odds ratio 2.678). Combinations of various parameters became significant when including free/total prostate-specific antigen, increasing prostate cancer detection to 88%. We conclude that free/total prostate-specific antigen is the most significant among prostate-specific antigen quantities (total age-dependent prostate-specific antigen, prostate-specific antigen density and prostate-specific antigen velocity). Adding this parameter to other prostate-specific antigen parameters improves the discrimination between prostate cancer and benign prostatic hypertrophy for the population at risk.

Synchronous and metachronous contralateral adrenal metastases from primary renal cell carcinoma.

Two rare cases of contralateral adrenal metastases are reported. One with a synchronous presentation where the renal tumor was removed together with a solitary large contralateral adrenal metastasis. The second patient underwent adrenalectomy for a metastatic lesion diagnosed 4 years after nephrectomy performed for renal cell carcinoma of the contralateral kidney. Five years following the adrenalectomy the patient underwent leg amputation because of a solitary bone metastasis.

Serum free/total prostatic-specific antigen in prostate cancer patients treated with LH-RH agonists.

OBJECTIVE: This study is based on promising results using the ratio of free/ total (F/T) prostatic-specific antigen (PSA) for discrimination between benign prostatic hypertrophy and prostate cancer. We tried to determine the value of F/T PSA in different clinical situations at a certain time point during follow-up of luteinizing-hormone-releasing hormone (LH-RH) agonist treatment and to correlate it to T-PSA. PATIENTS AND METHODS: 182 patients followed-up for different periods in the last 3 years were routinely monitored for serum T-PSA. During the last 11 months, F-PSA was also measured together with T-PSA, and the ratio of F/T PSA was calculated. In 26 patients, the ratio of F/T PSA was monitored sequentially in several samples. RESULTS: Although 5 patterns of clinical response to LH-RH agonists were identified according to previous T-PSA, the F/T ratio could significantly (p < 0.05) discriminate between patients responding to treatment in contrast to patients escaping, fluctuating or not responding to hormonal ablation. Those patients responding to hormones showed a higher F/T PSA ratio (36.5 +/- 33.1%) compared to the nonresponding group (12.0 +/- 10.1%). CONCLUSIONS: During individual follow-ups, the pattern of response to LH-RH treatment is reflected by the F/T PSA ratio: while successful treatment causes an F/T PSA increase, relapse is accompanied by a decrease in this ratio. However, the changes in the F/T PSA ratio did not precede the indicative changes in T-PSA. It seems that increased values of F/T PSA ratios are intrinsic features of 'benign' prostatic disease, and the molecular events resulting in different PSA molecules in various clinical situations have to be elucidated.

Single-dose intramuscular ketorolac versus diclofenac for pain management in renal colic.

A double-blind controlled study was designed to compare the effective- ness of a single intramuscular dose of 60 mg ketorolac with that of 75 mg diclofenac in the treatment of renal colic and to monitor side effects. Fifty-seven patients completed the study, 27 in the ketorolac group and 30 in the diclofenac group. Effectiveness of treatment was monitored by pain relief reported on a 4-point verbal scale at different time points. At 60 minutes 77.8% and 86.6% (P = 0.4) of patients, and at 120 minutes 81.5% and 96.6% (P = .1 5) of patients, reported significant pain relief following ketorolac and diclofenac doses, respectively. Both groups had an equal 92% significant pain relief at discharge from the emergency department. Both drugs were well tolerated by the patients. Ketorolac therefore, seems as effective as diclofenac in the treatment of renal colic.

A fibroepithelial urethral polyp protruding into the base of the bladder: sonographic diagnosis.

A case of congenital urethral polyp in a 10-month-old boy is presented. These rare congenital tumors can lead to urinary retention, hematuria, urinary tract infections, hydronephrosis and hydroureter. Traditionally the diagnosis was based either on voiding cystourethrography or cystoscopy. The sonographic features of these polyps have only been reported in a few cases in the past. This modality is not infrequently the imaging procedure of choice in the work-up of children with signs and symptoms related to the urinary system. In the case presented herein the polyp was demonstrated initially by sonography as a soft tissue mass arising at the base of the urinary bladder. The differential diagnosis of these lesions is presented.

Choriocarcinoma of the bladder. Report of a case of primary tumor or late metastasis of a molar pregnancy.

Choriocarcinoma of the bladder was diagnosed in a 54-year-old woman presenting with macroscopic hematuria 17 years following evacuation of a molar pregnancy. The patient was treated by cystoscopic transurethral tumor resection followed by three courses of triple-agent chemotherapy and total abdominal hysterectomy with bilateral salpingo-oophorectomy. Six months later a gradual rise in beta-human chorionic gonadotropin levels led to the diagnosis of recurrent bladder tumor at the site of the previous tumor. The patient underwent a conservative partial cystectomy, and 12 months postoperatively there was no evidence of disease. It is unclear whether the patient developed a primary urinary bladder choriocarcinoma or late metastatic disease from the previous molar pregnancy.

Identification and quantification of human kidney atrial natriuretic peptide receptors.

The present study determined 125I-label atrial natriuretic peptide (ANP) binding sites in human kidney glomerular and papillary membranes. The membranes were prepared from non-malignant renal tissue obtained at nephrectomy of patients with renal carcinoma. To evaluate the proportion of ANP receptor classes ANP-R1 (ANPR-A, -B) versus ANP-R2 (ANPR-C), competitive binding studies were performed using [125I]-ANP in the presence of increasing concentrations of ANP or an internally ring-deleted analog, des(Gln116, Ser117, Gly118, Leu119, Gly120)ANP(102-121), called C-ANP, which binds selectively to ANPR-C receptors. Analysis of the competitive binding curve with ANP in glomerular membranes suggested the presence of one group of high-affinity receptors with dissociation constant Kd = 26 +/- 12 pmol/l and density Bmax = 101 +/- 47 nmol/kg protein. A decrease of 10-30% in Bmax with no change in Kd was obtained in the presence of excess (10(-6) mol/l) C-ANP, suggesting the existence of a small amount of a second class of receptors, the ANPR-C class. The densities of ANPR-A, -B versus ANPR-C receptors in human glomeruli, calculated from competitive inhibition experiments, were 75 +/- 42 and 22 +/- 16 nmol/kg protein (N = 8). Autoradiography of the sodium dodecyl sulfate polyacrylamide gel electrophoresis under reducing conditions showed two bands: a highly labeled 130kD band and a weakly labeled 66 kD band, both displaced by ANP. Only the 66-kD band was displaced by the C-ANP analog. Human papilla membrane, as shown by competition binding studies and SDS gel electrophoresis, presented only one class of receptors with Kd = 40 +/- 23 pmol/l (mean +/- SD, N = 3) and Bmax = 17 +/- 6.3 nmol/kg protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Evaluation of prostate-specific antigen as a marker for adenocarcinoma of the prostate.

Serum prostate-specific antigen (PSA) concentrations were measured in samples from 437 subjects, including patients with prostate cancer, patients with benign prostate hyperplasia (BPH), and patients with genitourinary cancer or benign genitourinary diseases other than prostatic, as well as patients who had undergone prostatectomy and healthy controls. PSA concentration was elevated (less than 10 ng/ml) in 84.4% of the patients with prostate cancer, in 14.1% of the patients with BPH, and in 10% of the patients with genitourinary cancer (as compared with 2% of the patients with benign genitourinary diseases). PSA concentration was not elevated in any of the patients who had undergone prostatectomy nor in the controls. In the same samples, the level of prostatic acid phosphatase (PAP) was increased (greater than 4 IU/L) in only 55.6% of the patients with prostate cancer and in 7.4% of the patients with BPH. According to these findings, the sensitivity of the test is 84.4% (55.6% for PAP) and the specificity is 92.9% (94.9% for PAP). The increase in PSA concentration in BPH correlates well with the prostate mass (gamma = 0.794), although there was no patient with a prostate weight higher than 30 gm and a PSA concentration lower than 5 ng/ml, and all patients with PSA concentrations lower than 4 ng/ml had prostate weight of less than 10 gm.

Binding of serum prostate antigen to concanavalin A in patients with cancer or hyperplasia of the prostate.

The percentage of nonglycosylated prostate-specific antigen (PSA) was measured in the serum of 15 prostate cancer patients and 15 patients with benign hyperplasia of the prostate. The larger part of serum PSA in both groups was glycosylated, but while in carcinoma of the prostate the mean percentage of nonglycosylated PSA was 38.4 +/- 6.5, in benign prostate hyperplasia (BPH) only a mean of 14.2 +/- 4.3% of the PSA was nonglycosylated. These significantly higher results (p less than 0.001) suggest a different pattern of release of PSA from cancer cells and from hyperplastic or normal cells. Since in a part of the BPH we encounter elevations of PSA similar to the levels found in neoplasms, the degree of concanavalin A binding can provide an additional means in differentiating between benign and malignant lesions.

Adult nephroblastoma.

Clinical, pathologic, and ultrastructural features of two cases of adult nephroblastoma (Wilms' tumor) are presented, one in a 77-year-old man who had a large tumor with extracapsular extension and blood vessel invasion. Histologically, it consisted mainly of blastema. The patient died during the first course of postoperative chemotherapy with dactinomycin. The second patient was a 47-year-old woman with a tumor limited to the renal parenchyma. Histologically, the tumor was mainly epithelial. This tumor was discovered incidentally and remained unchanged for 5 years before surgery. The patient underwent nephrectomy and refused further treatment. She was free of disease 2 years after surgery.