Genotypic characterization of prostatic carcinomas: a combined cytogenetic, flow cytometry, and in situ DNA hybridization study.

Cytogenetic studies were performed on 36 biopsies obtained from 26 primary prostatic adenocarcinomas. Following histopathological characterization of control sections, the biopsies were investigated using metaphase cytogenetics, DNA flow cytometry, and fluorescence in situ DNA hybridization. In 12 specimens, no carcinoma was found in control sections by histopathological means. In 24 carcinoma biopsies clonal aberrations were detected in 15 specimens. Tetraploidy as sole aberration was detected in five specimens. Loss of the Y chromosome was seen in eight samples. Only one tumor revealed structural abnormalities. Eight samples were found to be normal (46,XY). Remarkably, nonclonal chromosome aberrations, particularly marked chromosome loss, were frequently detected in prostatic carcinomas and premalignant lesions (prostatic intraepithelial neoplasia). In the series of biopsies investigated by means of cytogenetics and flow cytometry, biopsies with aneuploid DNA content were found to be cytogenetically normal. Conversely, the cytogenetically aberrant clones were found to be of diploid DNA content. Evidence of focal intratumoral heterogeneity was revealed by cytogenetics, flow cytometry, and in situ hybridization.

Genetic changes and telomerase activity in human renal cell carcinoma.

Using the sensitive telomeric repeat amplification protocol assay, we detected telomerase activity in 26 of 35 (74.3%) renal cell carcinomas analyzed. Subdivision of the tumors according to telomerase activity did not reveal an obvious association between the presence of telomerase activity and histomorphological stage, grade, tumor size, or DNA ploidy. Furthermore, no association was found between telomerase activity and a distinct chromosomal aberration pattern; namely, loss of genetic material on the short arm of chromosome 3. Telomerase activity was also detected in 6 of 35 (17.1%) normal corresponding renal tissue samples, which seems interesting in light of the supposed biological role of telomerase expression in carcinogenesis. Interestingly, telomerase activity was detected in three of the four (75%) kidneys bearing non-clear cell tumor types, whereas of the 31 kidneys with clear cell carcinomas, telomerase activity was found in only 3 (9.7%) normal tissue samples. In addition, the two renal angiomyolipomas and one of the two analyzed transitional cell carcinomas of the renal pelvis were telomerase negative.

Two different forms of p53 localized differently within cells of urogenital tumours.

We analyzed the subcellular localization of p53 in prostate and bladder carcinoma cells. Using laser scanning microscopy and PAb1620, a monoclonal antibody recognizing the wildtype conformation of p53, and another monoclonal antibody directed against the mutant conformation of the protein (PAb240), we found two different subsets of p53 within the same cell. The wildtype subgroup was found in the nucleolus, whereas the mutant protein was confined to the nucleus. The results obtained by immunofluorescence were verified by Western blot analysis and immunoprecipitation. Thus, our findings demonstrate an unusual subcellular localization pattern of p53 in prostate and bladder cancer cells which may indicate another mechanism of inactivation of p53.

Phenotypic characterization of androgen receptor-related proliferation in prostatic-carcinoma derived cell-cultures.

Our study reviews the central role of androgens for the development of androgen-insensitive growth in prostatic carcinoma using an individual dynamic model which emulates aspects of proliferation control. Our aim is the identification of individually proliferating cells from prostate carcinoma. Short-term cultured cells (n=205) from radical prostatectomy specimens have been used as the major method to skew the correspondence between the androgen-receptor (AR), proliferation-markers (Ki-67, PCNA) and morphological landmarks (nucleoli, AgNOR). These techniques allowed the identification of distinct androgen-insensitive, proliferating cell-clusters and in the future will allow the analysis of genotypic changes in identified proliferating cell clones.

Different subnuclear localization of wild-type and mutant p53 in human prostate-cancer cells.

The growth suppressor protein p53 is abnormally expressed in a variety of different human tumor cells. We have analyzed the expression of p53 in cell cultures derived from tissues of radical prostatectomies and in the permanent prostate carcinoma cell line PC-3 using two different p53 specific monoclonal antibodies. With the wild-type specific monoclonal antibody PAb1620 we found p53 localized in nucleoli whereas only a few cells were positively stained in the nucleus with the mutant specific monoclonal antibody PAb240. Control experiments with p53 from SV80 cells which express wild-type p53 and HT29 cells expressing mutant p53 documented the specificity of the monoclonal antibodies. The specificity of the antibodies in recognizing indeed p53 was demonstrated further by immunoprecipitation analysis of p53 from the same cell cultures. Since p53 is usually localized to the nucleus our results may represent a specific feature of the wild-type phenotype of p53 in prostate carcinoma cells. The localization of p53 in nucleoli may be another mechanism of the inactivation of wild-type p53.

DNA aneuploidy in prostatic adenocarcinoma: a frequent event as shown by fluorescence in situ DNA hybridization.

Fluorescence in situ hybridization (FISH) using specific DNA probes for chromosomes 1, 7, 10, and Y was performed on 53 prostatic tissue samples obtained from 33 radical prostatectomy specimens and two benign control specimens. The 53 samples from carcinomatous prostates included 33 cancerous and 20 noncancerous samples. Additionally, four metastatic lymph node specimens were examined. Clonal chromosome abnormalities were observed in 78% of the tumors studied. They were detected in a higher proportion in stage pT2 and pT3 tumors (86% and 88%, respectively) compared with stage pT1 tumors (25%). No stage pT4 tumor was analyzed. There was evidence of remarkable focal intratumoral heterogeneity documented by the study of two samples from the same tumor in three of six cases. Comparing FISH determined ploidy patterns with DNA flow cytometry (FCM) in 22 samples, FISH showed aneuploidy whereas FCM showed none.

A new serial transfer explant cell culture system for human prostatic cancer tissues preventing selection toward diploid cells.

An improved explant cell culture technique to avoid selection of prostatic adenocarcinoma cells toward diploid cells is described. This method is based on 1) histologically characterized tissue explants, 2) the use of polyethylenteraphthalate (PET) membranes as growth surface, which are part of special inserts in six-well-plates to allow 3) cocultivation with heterologous fibroblasts, and 4) coating of the membranes with elements of the extracellular matrix. The main characteristic of this particular approach is the serial transfer of the tissue explant from one membrane to the other. Up to ten serial transfer steps could be performed to produce cell monolayers growing out of the same tissue specimen. Using this approach, 21 prostatic carcinoma specimens that were obtained from 13 primary prostatic adenocarcinomas after radical prostatectomy were cultivated. Ploidy of the cells was monitored by fluorescence in situ DNA hybridization using the centromere specific DNA probes pUC1.77, p alpha 7t1, and pY3.4. Interestingly, a high aneuploidy rate of the cell cultures was found with maintainance of aneuploidy in 18 (86%) of the 21 paraffin-embedded cancer tissue specimens with proved aneuploidy. Although a slight decrease of the proportion of aneuploid cells during serial transfer was observed, significant aneuploid cell populations were retained up to a maximum of ten transfer steps. These findings indicate that selection toward diploid cells can be prevented by improved cell culture techniques that mimic the in vivo situation.

Proteasome inhibitors and their combination with antiandrogens: effects on apoptosis, cellular proliferation and viability of prostatic adenocarcinoma cell cultures.

The 26S proteasome is a ubiquitin-dependent proteolytic system that has been implicated in the regulation of cell cycle progression and apoptosis. We investigated the effects of the proteasome inhibitors MG115 and PSI alone or in combination with different concentrations of the antiandrogen hydroxyflutamide on the cellular proliferation, apoptosis and viability of 10 prostatic adenocarcinoma cell cultures. Treatment with both proteasome inhibitors resulted in apoptosis induction, whereas the combinations with hydroxyflutamide generally did not, with the exception of MG115 combined with 10(-7) M hydroxyflutamide. MG115 caused a significant decrease in cellular proliferation, as did the combinations of both proteasome inhibitors with hydroxyflutamide, whereas hydroxyflutamide alone was only effective at a concentration of 10(-5) M. Cellular viability was significantly reduced when both proteasome inhibitors were combined with 10(-5) M hydroxyflutamide. Although the results varied among different cell lines, we conclude that proteasome inhibitors are able to induce apoptosis and reduce cellular proliferation. They might prove effective as antineoplastic substances in prostatic adenocarcinoma alone or in combination with antiandrogens.

[Computed tomographic studies of the scrotal contents, particularly the testis]. / Computertomographische Untersuchungen des Scrotalinhaltes, insbesondere des Hodens.

CT examination of the testes was carried out in 49 patients for the investigation of testicular tumours. Hypodensity and inhomogeneity were typical of teratomas and hyperdensity and relative homogeneity of seminomas. Granulomatous orchitis and lymphomas showed the same characteristics as seminomas. Three testes in the abdomen could be localised and classified. One non-palpable primary tumour was found as well as an old partially calcified tumour, three old torsions and one lipo-sarcoma. In about 10% it was not possible to distinguish between tumour and inflammatory lesions. Compared with sonography, CT has advantages, particularly in the diagnosis of old torsion.

[Surgical therapy of urogenital tuberculosis]. / Zur operativen Therapie der Urotuberkulose.

The surgical therapy of genitourinary tuberculosis is important as adjuvant treatment besides an appropriate anti-tuberculous drug therapy. During the years 1966-1983 surgical treatment was necessary in 46.1% (330 cases) of 715 patients. Cases, not treated properly for a long time, still have a high incidence of nephrectomy. Since 1976 more reconstructive operations and partial kidney resections have been carried out, resulting in a 5,6% rate of secondary nephrectomy. In up to 92.3% of the patients with these operations, renal function was not reduced or had improved. In cases with progressive tuberculosis of the upper urinary tract reconstructive surgical treatment is also recommended with good late results.

[Injuries of the urinary system and management in multiple trauma cases]. / Verletzungen des harnableitenden Systems und Management im Rahmen des Polytraumas.

Some of the most common lesions in patients with multiple traumas are injuries to the urinary system. A blunt trauma of the abdomen mainly leads to injuries of the kidney, ureter and/or bladder, whereas the external genitals are often damaged by contusions, decollements and even amputation. The multiply traumatized patient must necessarily be cared for and treated through an interdisciplinary cooperation. Thus, "urologisation" of the condition must be avoided, as must the non-recognition of urological injuries. Iatrogenic lesions are primarily observed in the context of urological, gynecological and surgical interventions. Immediate recognition may lead to early treatment, thereby avoiding possible, complications. The procedure for treating multiply traumatized patients will be shown using a plan of specific steps. Symptoms of injuries to the genitourinary system are usually not obvious. In most cases micro- and macro-hematuria are the leading symptoms, but they are not always demonstrable. In the acute phase the drainage of urine must be protected. It is mainly during the third step that urological lesions require surgical treatment. Catheterisation of the urethra may only be performed when the possibility of injury to it is excluded.

[Use of electronic data processing in the urologic clinic and practice--possibilities and perspectives]. / EDV-Einsatz in der urologischen Klinik und Praxis--Möglichkeiten und Perspektiven.

The development, present status and future trends in the use of computers in urology in the Federal Republic of Germany are reviewed. The hardware, software required for hospital and private practice and the staff needed are discussed. Proposals are given for the installation and stepwise upgrading of computer systems, from simple text processing units to complex hospital communication systems. Finally new technologies that might considerably change the use of computers in urology are presented.

[Electronic data processing-assisted text processing at the clinic and in general practice]. / EDV-gestützte Textverarbeitung in Klinik und Praxis.

Word processing is currently the most frequent application for personal computers, and a wide variety of standard software is available. The capabilities of modern word-processing software includes the convenient typing and correction of all routine correspondence, as well as the professional layout of scientific manuscripts. The decision to purchase a certain word-processing programm should be made according to local needs and prerequisities. Three to six months may be necessary to fully adapt the organization of a clinic or private practice to the new technology.

[Proteasome inhibitors: induction of apoptosis as new therapeutic option in prostate cancer]. / Proteasomeninhibitoren: Apoptoseinduktion als neue Therapieoption beim Prostatakarzinom?

New perspectives in prostate cancer genesis and putative clinical management have emerged in recent years . Apoptosis plays a major role in this environment. Proteasome inhibitors block the action of a multicatalytic proteinase complex involved in the degradation of intracellular proteins, particularly with regard to cell cycle regulation and apoptosis. Numerous in vitro studies have demonstrated the ability of these compounds to induce apoptosis and enhance the activity of conventional tumoricidal agents in many cancer cell types, including prostate cancer cells. They point out the use of these potent inhibitors as a new potential molecular approach to the therapeutic management of prostate cancer. Furthermore, the action of proteasome inhibitors has been tested in animal models and in patients with hormone refractory prostate cancer, resulting in both PSA and tumor volume decrease. PS-341 (bortezomib, Velcade) is the first proteasome inhibitor with clinical application in cancer therapy that has been used in clinical trials to date. This report reviews the current status of those papers that have tried to analyze the connection between the proteasome pathway and apoptosis. We present our results of proteasome inhibition in individual prostate cancer cell lines. Proteasomal inhibition may offer a new therapeutic access in "molecular targeting" of prostate cancer.

[Retroperitoneal extragonadal germinal choriocarcinoma: description of a case and the complex therapeutic approach]. / Chorio-carcinome germinal extragonadique à localisation rétropéritonéale: description d'un cas et de sa conception thérapeutique complexe.

Patients with primary extragonadal, retroperitoneal germ cell tumors usually have very advanced disease and present uncharacteristic signs and symptoms. We report a case of a patient with large retroperitoneal choriocarcinoma. After four initial cycles of chemotherapy, the patient underwent debulking surgery including angiorrhaphy and-plasty. This therapeutic management was justified, as it revealed active residual tumor. Furthermore, a large tumor mass was removed with vascular and ureteric decompression. As a result of this treatment, adjuvant chemotherapy was more effective. In conclusion, despite possible complications, we recommend this regimen in young patients.