Analysis of 7 immunohistochemical markers in male germ cell tumors demonstrates the prognostic significance of p53 and MIB-1.

BACKGROUND: Various prognostic factors have been investigated in order to predict the minority of male germ cell tumor (GCT) patients who will develop resistant disease. However, no prognostic system has been proven accurate. MATERIALS AND METHODS: Paraffin-embedded tissue specimens, obtained from primary lesions during the initial diagnosis of 83 advanced chemotherapy-treated GCT male patients, were stained for 7 immunohistochemical markers: p53, bax, bcl-2, MIB-1, topoisomerase IIa, c-kit and COX-2. The percentage of positive cells for each marker was measured for each patient. Cox regression was used for the prognostic factor analysis. RESULTS: All patients were followed for a median of 4 years. Nineteen patients had seminoma and 64 non-seminomatous GCT. In univariate analysis, only p53 (hazard ratio (HR) = 4.01, 95% confidence interval (CI) = 1.25-12.84, p = 0.019) and MIB-1 (HR = 3.16, 95% CI = 1.06-9.45, p = 0.039) were found to be prognostic for disease-specific survival. The best prognostic cut-off values of p53 and MIB-1 were 10% and 30% respectively. In multivariate analysis, these two markers obtained independent significance only when considered in combination (HR = 6.63, 95% CI = 1.40-31.41, p = 0.017, for patients with one or both markers above their cut-off), while the International Germ Cell Consensus Cancer Group (IGCCCG) risk was the most significant (HR = 7.99, 95% CI = 1.96-32.52, p = 0.004, for the high-risk group). However, the expression of these markers seemed to be significantly correlated with known prognostic factors. Nevertheless, we identified 34 patients of low IGCCCG risk expressing both markers below their cut-off with excellent survival. CONCLUSION: Among 7 immunohistochemical markers, p53 and MIB-1 demonstrated prognostic significance. Their combination may contribute to improvement of the accuracy of the currently approved prognostic system (IGCCCG).

Microvessel density (MVD) and cyclooxygenase-2 (COX-2)/ beta-catenin interaction are associated with relapse in patients with transitional carcinoma receiving adjuvant chemotherapy with paclitaxel/carboplatin: a hellenic cooperative oncology group (HECOG) study.

BACKGROUND: Cycloxygenase (COX)-2 has been associated with proliferation, apoptosis and angiogenesis in urothelial cancer. The prognostic significance of COX-2 in patients who received adjuvant chemotherapy for urothelial cancer was examined. PATIENTS AND METHODS: Expression of COX-2, p53, ki67, beta-catenin, vascular endothelial growth factor (VEGF) and microvessel density (MVD) were studied retrospectively in 59 patients with urothelial cancer (pT3, pT4, N+) who had undergone surgery. The patients had subsequently received adjuvant chemotherapy. RESULTS: Thirty-eight out of 59 cases (64%) were positive for COX-2. COX-2 was not associated either with progression-free survival (PFS) or overall survival (OS). MVD levels > or =47 were associated with longer median PFS compared with lower levels (not reached vs. 13 months [95% CI: 8-18], p=0.048). The median PFS for patients with beta-catenin nuclear accumulation and COX-2 expression was 6 months (95% CI: 4-7) compared with 19 months (95% CI: 14-23) for neither or only one of these factors (p=0.018). CONCLUSION: MVD may be a useful indicator of relapse in high-risk urothelial cancer treated with adjuvant chemotherapy.

Kit expression in male germ cell tumors.

OBJECTIVE: KIT functions as the receptor for stem cell factor (SCF) and this interaction is essential for regulation of proliferation and survival, particularly for germ cells since it regulates oogenesis, folliculogenesis and spermatogenesis. Up-regulation of KIT signalling has been associated with oncogenic transformation in cells expressing the molecule. Our objective was to investigate the expression of KIT in germ cell tumor patients and correlate it with the patients' clinical characteristics. PATIENTS AND METHODS: One hundred and seventy-three archival blocks of formalin fixed, paraffin-embedded tumor samples from histologically confirmed germ cell tumor (GCT) patients were included in the study. Immunohistochemical staining for KIT was performed and the percentage of positive cells was calculated by an independent pathologist. KIT expression was considered as positive if > 10% of tumor cells displayed membranous or cytoplasmic staining. RESULTS: Sixty-one patients with seminomatous (49 pure, 11 anaplastic, 1 spermocytic) and 112 with non-seminomatous GCTs (36 malignant teratoma undifferentiated (MTU), 15 malignant teratoma trophoblastic (MTT), 20 malignant teratoma intermediate (MTI), 35 malignant teratoma combined (MTC) and six others) were identified. Among pure seminoma patients, 38 (77.5%) revealed a positive staining for KIT, while only two out of eleven (18.2%) anaplastic seminoma patients were identified as positive. This difference was statistically significant (p < 0.001). Among 35 patients with an MTC, 48.6% had a positive KIT staining while only two of the remaining patients with a non-seminomatous GCT had a positive staining. Although KIT was strongly correlated with seminomatous histology (p < 0.001), it failed to correlate with stage (p = 0.19) or treatment response (p = 0.11) in these patients. Overall, four (one seminoma, three MTC) out of 22 chemoresistant patients showed a positive staining for KIT. CONCLUSION: KIT is expressed in the majority of seminomas and in seminomatous components of the combined tumors, but only in a minority of anaplastic seminomas and rarely in non-seminomatous GCTs. The recent development of tyrosine kinase inhibitors may offer a possibility of cure in chemoresistant patients overexpressing KIT.

Prognostic significance of CA 125, CD44, and epithelial membrane antigen in renal cell carcinoma.

OBJECTIVES: To study retrospectively CA 125, CD44, and epithelial membrane antigen (EMA) expression in renal cell carcinoma and their role as prognostic factors. CD44 is a cell adhesion molecule, and CA 125 and EMA are tumor-associated antigens used in the diagnosis and monitoring of the outcome and response to treatment of various human malignancies. Their expression and prognostic significance after resection of renal cell carcinoma have not been adequately studied. METHODS: The expression of CA 125, CD44, and EMA were studied immunohistochemically and correlated with the outcome of 92 patients who underwent nephrectomy for renal cell carcinoma. RESULTS: Positive staining was found for CA 125 in 28 patients (30.43%), CD44 in 48 patients (52.17%), and EMA in 74 patients (80.43%). CA 125 expression was increased in those with higher T stage (P <0.001) and histologic grade (P = 0.007). An inverse relationship was found between EMA expression and grade (P <0.001). The median follow-up was 41.5 months (range 30 to 65). The median survival for positive and negative patients was 34.6 versus 54.3 months for CA 125 (P = 0.0044), 48.3 versus 51.5 months for CD44 (P = 0.4677), and 53.2 versus 34 months for EMA (P = 0.0046). Multivariate analysis showed that CA 125 and EMA expression were independent prognostic factors (P = 0.021 and P = 0.018, respectively). Subgroup analysis showed that CA 125 expression predicted a significantly higher probability of death (28.6% versus 8%, P =0.0413) in patients with T1 or T2 tumors. CONCLUSIONS: CA 125 and EMA appear to be useful prognostic markers in renal cell carcinoma. Additional studies are needed to determine the value of these markers as a means of selection for postoperative management.

Deoxyribonucleic acid flow cytometry in the assessment of spermatogenesis.

PURPOSE: We compared deoxyribonucleic acid (DNA) flow cytometric analysis of testicular tissue to quantitative assessment of spermatogenesis. MATERIALS AND METHODS: We studied 35 infertile men with azoospermia or oligospermia. All patients underwent incisional testicular biopsies. DNA flow cytometric analysis was performed on each specimen to evaluate the ability of the method to quantify alterations in spermatogenesis. The results were compared to quantitative histological examination. At least 100 spermatic tubules were examined on each specimen and the number of spermatids per tubule was counted. All histological specimens were examined by the same pathologist. RESULTS: Of the 35 specimens analyzed with DNA flow cytometry 5 were normal, while the percentage of haploid cells (spermatids and spermatozoa) was decreased (hypospermatogenesis) in 14, complete maturation arrest was noted in 2 and almost complete absence of haploid cells was found in 14. Comparing the findings on histological examination with histograms, excellent correlation was noted in cases of the Sertoli-cell-only syndrome and complete maturation arrest, while 3 of 14 histograms with hypospermatogenesis demonstrated normal spermatogenesis on histological examination. Additionally 1 of 5 histograms with norma, spermatogenesis demonstrated hypospermatogenesis on histological examination. CONCLUSIONS: DNA flow cytometry of the testicular tissue seems to be an objective and quantified method that can be used to investigate spermatogenesis in infertile men. It is also less time-consuming than any histological examination, permits management decisions within 1.5 hours after biopsy and may replace testicular histopathological study. Flow cytometric diagnoses correlated well with histopathological findings.

Sodium thiosulfate in the treatment of tumoral calcifications in a hemodialysis patient without hyperparathyroidism.

Extraskeletal tumoral calcifications (TC) may occur in patients with end-stage renal disease. The TC usually develop in the presence of secondary hyperparathyroidism or a high calcium x phosphate product, while other factors have been also occasionally implicated in their development. At present, no uniformly accepted effective treatment has been described for this condition. We describe a 58-year-old female patient with end-stage renal disease who 7 years after the onset of dialysis presented with pain and movement restriction of various joints. A skeletal X-ray showed huge amounts of periarticular TC. The TC occurred in the absence of hyperparathyroidism or a high calcium x phosphate product as evidenced by hormonal and biochemical examination as well as by a bone biopsy specimen that revealed an adynamic bone disease with significant aluminum staining. Sodium thiosulfate, an inorganic salt that has been claimed to inhibit the formation and to favor the solubility and the mobilization of calcified masses, was administered to the patient, and after a long period of treatment considerable radiological regression of the TC with concurrent clinical recovery was noticed. Aluminum intoxication, along with other factors, was considered to be the cause of TC development. The use of sodium thiosulfate seemed to be a reasonable nonspecific therapeutic approach for the management of TC in this case.

Transurethral thermotherapy in the management of benign prostatic hyperplasia.

Transurethral radiofrequency thermotherapy for symptomatic benign prostatic hyperplasia was performed in 50 selected patients, using the THERMEX II device. High surgical risk patients were included, among them 13 previously catheterized ones, because of unresolved retention for more than 6 months. The treatment consisted of a three-hour single session at 47 degrees C. Follow-up studies were carried out at 1, 3, 6 and 12 months using Madsen score, maximum flow rate, residual urine volume determinations and prostate bulk measurement by transrectal ultrasound. Subjective and objective improvement (of more than two parameters) was noticed in 62% of the patients. Prostatic volume did not change. In the retention group 54% of the patients remained free of catheter. Postoperative histology in 8 cases that failed to respond, revealed focal haemorrhagic and necrotic changes in periurethral glandular tissue. This is a promising method for selected high risk patients that achieves reasonable but not comparable results to TURP and requires further investigation in larger patient groups with BPH.

Bladder washing flow cytometry assessment of nephrogenic adenoma of the bladder.

A case of nephrogenic adenoma of the bladder is reported. This rare benign lesion that histologically resembles primitive renal collecting tubules is considered to represent a metaplastic response of urothelium to several stimulating factors. DNA flow cytometry performed on preoperative bladder washing revealed an aneuploid phenotype.

The value of neuroendocrine markers in non-small cell lung cancer: a comparative immunohistopathologic study.

In order to estimate the value of immunohistochemical identification of neuroendocrine (NE) differentiation markers in non-small cell lung carcinomas (NSCLCs), we investigated the expression of five neuroendocrine and neural differentiation-related antigens in 51 NSCLCs. Additionally, 20 epithelial lung tumors with NE differentiation [15 carcinoids and five small cell lung carcinomas (SCLCs)] and 61 epithelial tumors of various other origin (breast, prostate, colon and head-neck carcinomas) were studied. An indirect two-stage immunoperoxidase method was performed in formalin-fixed and paraffin-embedded tissue specimens, by using commercially available monoclonal antibodies. These antibodies are directed against neuron-specific enolase (NSE), chromogranin-A and Leu-7 which are general markers of NE differentiation, bombesin, which is a specific NE secretory product and neurofilament triplet protein (NFTP), an intermediate filament protein of neuronal differentiation. All five markers demonstrated a positive immunoreactivity in NSCLCs, equally distributed to all three histologic subtypes, ranging from 16 to 47% of the cases (NSE 47%, bombesin 21.5%, Leu-7 21.5%, chromogranin-A 18% and NFTP 16%). Most of the carcinoids and SCLCs expressed multiple or all NE markers. The other four epithelial tumors showed a positive immunoreactivity for bombesin, Leu-7 and NFTP, ranging from 11 to 40% of the cases. Chromogranin-A was not expressed in any of these tumors, whereas NSE was demonstrated only in 17% of breast carcinomas. The following remarks can be drawn from this study: (1) some NSCLCs showed immunophenotypic NE differentiation; (2) among all the markers used, NSE was the most sensitive (sensitivity, 100%) and chromogranin-A the most specific (specificity, 100%); and (3) NSE and chromogranin-A appear to be the most valuable and useful indicators of probable neuroendocrine differentiation in lung epithelial tumors.

Primary localized amyloidosis of the urinary bladder.

A case is presented of a woman aged 78 with a primary location of amyloidosis of the bladder. The clinical symptoms and the cystoscopy initially suggested a vesical carcinomatous infiltration.

Immunohistochemical and immunocytochemical study of bladder carcinomas using the epithelium-specific, tumour-associated monoclonal antibodies HMFG1 and AUA1.

The antigenic expression of normal bladder epithelium and transitional carcinomas has been studied using the epithelium-specific, tumour-associated monoclonal antibodies HMFG1 and AUA1. Tissues from 79 cases of bladder carcinoma and 11 cases of non-neoplastic bladder tissues were stained with both the haematoxylin-eosin (H/E) and the indirect two-stage immunoperoxidase methods using the monoclonal antibodies (MAbs) HMFG1 and AUA1 at a concentration of 25 micrograms ml-1. Positive and negative controls were also used. Moreover, 46 urine smears prepared after cytocentrifugation were stained with both the Papanicolaou and the indirect two-stage immunoperoxidase methods. The results showed that HMFG1 reacted with the majority of cases of grade III carcinomas and carcinomas in situ and with a subset only of low-grade (I and II) carcinomas. The pattern of staining showed the following characteristics: (1) the epithelial surface membrane stained both in normal bladder and bladder carcinomas (surface of the papillae), (2) a variant number of cancer cells, increasing with the degree of malignancy, showed membrane and/or cytoplasmic staining, (3) tumours of the same histological grade showed antigenic heterogeneity. The MAb AUA1 was not widely expressed. The immunocytochemical study confirmed the reaction of HMFG1 with a variant number of malignant urothelial cells exfoliated in urine. Their reaction with AUA1 was much more limited. The immunocytochemical staining seemed to be more sensitive in the detection of malignant cells in some cases which had been characterized as negative or suspicious for malignancy by the Papanicolaou examination. The intravesical treatment with chemotherapeutic agents did not seem to influence the antigenic expression of malignant urothelial cells.