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W F Feitz

Publications and source records attributed to W F Feitz.

At least 37 records · Page 2Linked to original sources
PubMed1987

Tissue-specific markers in flow cytometry of urological cancers. III. Comparing chromosomal and flow cytometric DNA analysis of bladder tumors.

Thirty-seven transitional-cell carcinomas (TCC) of the urinary bladder were analyzed by DNA flow cytometry (FCM). After labelling of the cell suspensions with antibodies to cytokeratin, the cytokeratin-positive cells and the non-epithelial cytokeratin-negative cells could be analyzed separately. After estimation of S- and G2M phase, 3/17 cases (18%) with a normal DNA index showed elevated proliferative levels, among cytokeratin-labelled suspensions only. Of these 17 cases, 14 showed chromosomal abnormalities. The remaining 20 cases were abnormal, irrespective of the technique used. Although immuno-labeling of tumor cells for cytokeratin in FCM increases the sensitivity of this method in detecting aneuploid tumors or tumors with high proliferation fractions, the discriminating power of chromosomal analysis of TCC is greater than FCM.

Carcinoma, Transitional Cell↗
PubMed1987

Comparison of tissue disaggregation techniques of transitional cell bladder carcinomas for flow cytometry and chromosomal analysis.

DNA index (DI) measurements and chromosomal analysis of 42 transitional cell carcinomas were done after mechanical and enzymatical disaggregation of the tumor specimens. The results obtained with these different disaggregation techniques were compared in the 33 cases (79%) that showed recognizable chromosomes. The enzymatically obtained cell suspensions could not be used for chromosomal analysis after short-term culture of 24 hours. In four cases, the DI after enzymatical treatment could not be estimated. In most cases, the DI obtained from the tumor cells was similar for both aggregation techniques, with the exception of four cases of enzymatically treated cell suspensions in which the DI could not be estimated. The average DI of the aneuploid tumors was 13% higher than the corresponding chromosome count. In 19% of the aneuploid tumors the proportion of aneuploid cells could not be measured after enzymatical treatment. In the remaining suspensions the proportion of diploid cells was higher after enzymatical disaggregation than after mechanical treatment. It is concluded that for flow cytometric and direct chromosomal analysis of bladder tumors, the mechanical disaggregation technique is most suitable.

Adult↗
PubMed1987

Expression of proto-oncogenes in xenografts of human renal cell carcinomas.

In a recent paper, we described the expression pattern of proto-oncogenes in primary human renal cell carcinoma [12]. To test the possibility of using xenografts as a useful alternative for such studies, we analyzed xenografts of a number of human renal cell carcinomas in nu/nu mice. Xenografts included RC2, RC14, RC21, RC43 and NC65. Northern blot analysis indicated that c-Ras was expressed in all these xenografts. The identity of the ras transcripts in the individual xenografts was further specified as c-Ha-ras, c-Ki-ras or N-ras. Expression of c-myc and the p53 gene was also found in a number of these tumors. Only RC21 failed to express the c-myc or the p53 gene. In all xenografts, a 3.0 kb c-fes/fps mRNA was present. In RC2, RC14, RC21 and RC43, low levels of the 4.8 kb ab 1 transcript were detectable. Transcripts of myb and sis could not be detected in any of the xenografts. The results indicated that the expression pattern of a variety of proto-oncogenes in xenografts of human renal cell carcinomas was similar to that in the primary tumors.

Animals↗
PubMed1987

Intermediate filament proteins as tissue specific markers in normal and neoplastic testicular tissue.

Normal testicular tissue and primary and metastatic testicular germ cell tumours were examined for their intermediate filament protein (IFP) expression. Seminomas were shown to react with antibodies to vimentin, while non-seminomatous germ cell tumours were strongly positive with antibodies to cytokeratin. In the case of teratocarcinoma, several components of the tumour can be distinguished using a combination of monoclonal and polyclonal antisera in the double-label immunofluorescence technique. We conclude that antibodies to cytokeratin and vimentin can be helpful in the diagnosis of testicular germ cell tumours, especially in the differentiation between seminomas and non-seminomatous testis tumours.

Fluorescent Antibody Technique↗
PubMed1986

Tissue-specific markers in flow cytometry of urological cancers. II. Cytokeratin and vimentin in renal-cell tumors.

Nine primary human renal-cell tumors (RCT), one lymph-node metastasis, 4 human xenografts of a RCT in nude mice and a rat RCT line were analyzed by flow cytometry (FCM) using propidium iodide for DNA analysis and antibodies to cytokeratin and vimentin in the indirect immunofluorescence technique for labelling of specific tumor-cell populations. By means of 2-dimensional FCM analysis, vimentin- and cytokeratin-positive (tumor) cells were compared and their DNA content and proliferative fraction analyzed separately from those of cytokeratin-negative stromal and inflammatory cells. In primary human RCT, 2 subpopulations of cells were detected and analyzed separately. Small numbers of tumor cells with an abnormal DNA stemline were also detected. In addition, co-expression of intermediate filament proteins of both the cytokeratin and the vimentin types was detected in the aneuploid cell population. Comparison of 2 model systems of RCT with primary human RCT revealed a similar pattern of tumor-cell subfractions within these tumors. The 2-parameter FCM analysis permits the detection of subpopulations in complex cell suspensions and the quantification of these fractions, as well as analysis of their cellular DNA content.

Animals↗
PubMed1986

Establishment and characterization of seven Dunning rat prostatic cancer cell lines and their use in developing methods for predicting metastatic abilities of prostatic cancers.

In vitro cell lines were established from seven biologically distinct in vivo Dunning R3327 rat prostatic tumor sublines. Some of these in vitro cell lines (i.e., G, AT-1, AT-2) retain a low metastatic ability when inoculated back into syngeneic Copenhagen male rats, while others (i.e., AT-3, MAT-LyLu, MAT-Lu) retain a very high metastatic ability. A series of genetic (i.e., DNA content per cell, modal chromosomal number), as well as phenotypic parameters (i.e., morphology, 5 alpha-reductase, androgen receptor, estrogen receptor) were used to validate that the in vitro cell lines retained the major characteristics of the parental in vivo tumor sublines used for their respective establishment. A series of additional characteristics (i.e., morphology, growth rate, saturation density in surface culture, anchorage-dependent and -independent clonogenic potential) were compared between the high vs. the low metastatic in vitro cell lines to determine if a discriminatory parameter could be identified which reproducibly predicted the metastatic abilities of the particular prostatic cancer cell line. While the combination of the in vitro cell lines and their parental in vivo tumor subline will be a valuable tool for developing methods for predicting metastatic ability of prostate cancers, no single parameter yet measured is entirely successful in making this important distinction.

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗
PubMed1986

Expression of the human fes cellular oncogene in renal cell tumors.

Renal cell tumors were screened for expression of the cellular oncogenes c-abl, c-fes, c-fms, c-myc, c-ras, and c-sis in dot blot hybridization analysis. Expression of c-ras and c-myc was clearly detectable in most of the 15 tumors that were studied. The c-fes oncogene appeared to be expressed in only two of them. Comparative Southern blot analysis of molecularly cloned human c-fes DNA and genomic DNA of the 15 renal tumors revealed no major genetic differences. Northern blot analysis of poly(A)-selected RNA from the fes-positive tumors with the complete viral v-fes oncogene of the Gardner-Arnstein strain of feline sarcoma virus as a molecular probe revealed hybridization of RNA species of 3.0 and 4.5 kb, respectively. The 3.0 kb c-fes transcript has also been reported in RNA from patients suffering from acute myelogenous leukemia. The 4.5 kb transcript, however, has not been described before and represents either a c-fes-related splicing intermediate or, more likely, a completely processed transcript. The results of this study could imply that human c-fes coding sequences are more extensive than was previously assumed.

Carcinoma, Renal Cell↗
PubMed1986

Dynamics of human renal tumor colony growth in vitro.

Two-layer soft agar cultures from 26 patients with renal cell carcinoma, 21 renal primary lesions and 5 metastatic lesions, were evaluated for tumor colony formation using both dynamic growth curves and static single time point colony counting. Dynamic growth curves markedly increased the number of evaluable tumor cultures. There was no relationship between colony formation and TNM stage of the tumor or renal vein invasion. However, there was a significantly (p less than 0.02) higher rate of colony formation from younger (less than 50 y.) patients than from older patients. Only 2 of 5 tumors tested showed response to one or more cytostatic agents in vitro, both tumors showing response to Adriamycin and one to Cis-platin. Dynamic evaluation of tumor colony formation in soft agar may increase the clinical applicability of the human tumor cloning system both by increasing the number of evaluable cultures and by providing more information about the processes involved in tumor colony formation in vitro.

Adult↗
PubMed1986

The effects of intravesical and intradermal application of a new B.C.G. on the dog bladder.

Intravesical and intradermal application of B.C.G. (Bacillus Calmette Guérin) has proven to be effective in the prophylaxis of recurrence of superficial bladder carcinoma after transurethral resection and in the treatment of carcinoma in situ (C.I.S.) in man. Different strains of B.C.G. have been used for this purpose. In this article a new strain of B.C.G. (B.C.G.-R.I.V.M.) has been tested to assess its toxicity. The effects of intravesical and intradermal application of B.C.G.-R.I.V.M. were studied on normal and on coagulated canine urothelium. In this study no general side effects of B.C.G.-R.I.V.M. were seen. Only minor local changes occurred in the bladder wall. Small granulomas were found in the suburothelial tissue. No granulomas or signs of active inflammation were observed in the pelvic lymphnodes, spleen and liver. Because B.C.G.-R.I.V.M. seemed to be a safe agent in the dog we have started to use it for prophylaxis in superficial bladder cancer in man.

Administration, Intravesical↗
PubMed1986

Intermediate filament proteins as tissue specific markers in normal and malignant urological tissues.

Immunocytochemical techniques have become valuable tools in many fields of clinical pathology and medical research. Especially the development of highly specific (monoclonal) antibodies to a large variety of tissue antigens has in recent years led to the establishment of sensitive tissue markers. One of the most promising types of tissue specific markers so far is represented by the intermediate filament proteins. Since the findings of this rapidly expanding field are also being applied in urology, we have reviewed the current data in order to describe the new insights in tumor biology and histogenesis, as well as their application in diagnostic pathology.

Animals↗
PubMed1986

Application of antibodies to intermediate filament proteins as tissue-specific probes in the flow cytometric analysis of complex tumors.

The flow cytometric (FCM) analysis of carcinomas is often hampered by the presence of stromal and inflammatory cells in the cell suspensions obtained from such neoplasms. Therefore, an FCM method was developed to distinguish epithelial from nonepithelial cells by using polyclonal and monoclonal antibodies to (cyto)keratins, the epithelial type of intermediate filament proteins. Using a model system of cultured bladder carcinoma (T24) and leukemia (MOLT-4) cells, we tested our hypothesis and procedures by labeling cell mixtures with these antibodies. After incubation with an appropriate intermediate filament antibody and propidium iodide staining, the DNA content and distribution of T24 cells could be analyzed separately from MOLT-4 cells. When applied to cell suspensions of endometrial carcinomas, bladder carcinomas and Grawitz tumors, only the epithelial (primarily carcinoma) cells were stained for cytokeratin; these cells could thus be analyzed separately from stromal, inflammatory and other nonepithelial cells. In this way, a more accurate FCM analysis of the malignant fraction within a tumor can be achieved.

Adenocarcinoma↗
PubMed1985

Tissue-specific markers in flow cytometry of urological cancers: cytokeratins in bladder carcinoma.

Thirty-eight transitional-cell carcinomas (TCC) were analyzed by flow cytometry (FCM) using propidium iodide for DNA analysis and antibodies to cytokeratin by indirect immunofluorescence. By means of two-dimensional FCM analysis, cytokeratin-positive tumor cells could be analysed separately from cytokeratin-negative stromal and inflammatory cells. This resulted in an 18% increase in sensitivity of FCM detection of aneuploidy (10/38 samples with one-parameter DNA analysis versus 15/38 samples with two-parameter DNA and cytokeratin analysis). In addition, S-phase could be determined in the 15 aneuploid samples by means of two-parameter analysis where this was not possible using only DNA content because of the overlap of diploid and aneuploid populations. FCM analysis allowed quantification of the percentage of tumor cells expressing cytokeratin 18 which has previously been shown to correlate quantitatively with higher grade, higher stage TCC. The quantitative measurement of tumor-cell expression of cytokeratin 18 by FCM analysis appears to provide additional information of potential prognostic value, independent of tumor-cell ploidy and proliferative fractions.

DNA↗
PubMed1985

Cell DNA content--correlation with clonogenicity in the human tumour cloning system (HTCS).

Thirty-six ovarian and renal-cell tumours were analyzed by flow cytometry (FCM) for DNA content and in parallel were assayed for colony formation in a human tumour cloning system (HTCS). While 15/19 (79%) tumours with an abnormal (aneuploid) DNA stemline formed colonies in the HTCS, only 2/17 (12%) of diploid tumours formed colonies. All samples contained tumour cells as assessed by routine cytological examination. The capacity to form colonies in the HTCS was not correlated in these tumours with grade or stage of disease or tumour type. The level of aneuploidy expressed as the FCM DNA index did not correlate with the cloning efficiency in HTCS. These findings suggest that tumour growth in the HTCS reflects a biologically important potential, related in at least some tumours to an abnormal DNA stemline.

Carcinoma, Renal Cell↗
PubMed1985

Automated counting of human tumour colonies in the Courtenay-Mills assay system.

A procedure for using the Omnicon automated image analysis system for counting colonies grown from a human tumour cell line (COLO 205) in the Courtenay-Mills assay is described. This involves the transfer of the agar medium from culture tubes into petri dishes. Comparisons of observer and instrument counts were done on a blinded basis. Run-to-run correlation coefficient was 0.996 for automated counting and the inter-observer correlation coefficient was 0.984. Both assessments showed a linear relationship between the number of cells plated and the number of colonies grown. Automated colony counting is fast, reliable and provides additional information on colony size distribution, not obtainable with manual counting. This automated procedure will greatly facilitate in vitro drug sensitivity evaluation.

Autoanalysis↗
PubMed1985

Time course of ovarian tumour growth in soft agar culture.

Single time point assessment is usually employed in the Human Tumour Cloning System as the only parameter for in vitro growth. This does not seem to give a fair expression of the dynamic biological properties of tumour growth and time dependent effects, e.g. of cytotoxic drugs. We studied the time course of colony formation in temporal growth patterns (TGPs) and compared this method of growth evaluation with conventional single time point assessment in 57 samples of ovarian tumour cultures in the HTCS. A first advantage of the use of TGPs is that more cultures become evaluable, as this assessment over time can detect a rise in the number of colonies in dishes where colony-like clumps have initially been seeded. Thus only 28 of the cultures were evaluable for single time point assessment, whereas 57 were available for TGP evaluation. Growth was more often seen at TGP evaluation (14/57) than at single day assessment (8/57). Evaluation of growth over the course of time potentially allows detection of sensitivity to drugs. Furthermore TGPs reflect the dynamics of biological growth. These features cannot be studied in single time point assessment.

Agar↗
PubMedDate not supplied

Early urethral obstruction sequence: a lethal entity?

Early urethral obstruction sequence (EUOS) is characterized by severe bladder distension by the end of the first trimester of pregnancy, resulting in renal dysplasia or hydronephrosis, oligohydramnios and subsequent lung hypoplasia. We reviewed the outcome of 18 fetuses with suspected EUOS. The mean gestational age at the time of diagnosis was 15 weeks. In 1 fetus, a vesicoamniotic shunt was placed unsuccessfully. Pregnancy was terminated in 11 fetuses and 9 showed signs of pulmonary hypoplasia and severe renal disease on autopsy. In 3 cases, the autopsy material could be evaluated. Premature delivery occurred in 2 patients, and 5 chose to deliver at term. All 7 fetuses died soon after birth because of respiratory failure and lung hypoplasia; renal abnormalities were also found on autopsy. Since it is currently not clear whether prenatal intervention will prevent renal dysplasia and it is not known how early it should be done to prevent pulmonary hypoplasia, we believe that termination of pregnancy should be discussed when EUOS is suspected.

Adult↗
PubMedDate not supplied

Multiparameter analysis of four human renal cell carcinoma xenografts in nude mice.

Four human renal cell carcinoma xenografts (RC2, RC14, RC43, NC65), maintained in nude mice for several years, were investigated in a multi - disciplinary study, using (immuno) histochemical, biochemical and ultrastructural techniques. Histological, cellular, nuclear and biological characteristics were investigated. All tumors showed histologically recognizable features of human renal cell carcinomas, although marked differences between the four tumors were seen, both at the histological and ultrastructural level. Flowcytometric analysis of tumor cell suspensions allowed DNA quantification as well as the detection of subpopulations. Immunohistochemical staining procedures using tissue specific antibodies against intermediate filament proteins revealed two populations of tumor cells. Most tumor cells in three of the xenografts coexpressed cytokeratins and vimentin, while in RC43 most of the tumor cells expressed only vimentin. Northern blot analysis showed a higher expression of vimentin mRNA in all tumors as compared to normal kidney tissue. RC43 showed a three-fold higher level of vimentin mRNA than the other xenografts. Growth potential in the human tumor cloning system was evaluated by temporal growth pattern analysis. These experiments showed that the xenografts resemble human primary renal cell tumors in different ways, and reflect different characteristics that can be present in human renal cell carcinoma.

Aneuploidy↗