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Biomedical subjects

C J Herman

Publications and source records attributed to C J Herman.

At least 55 records · Page 3Linked to original sources
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

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

A computer model of spermatogenesis in the rat; correlation with flow cytometric data based on autoradiographic cell-cycle properties.

A computer model of rat spermatogenesis was created, based on autoradiographic studies of durations of the phases of the cell cycle (G1, S, G2 and mitotic phases) of each germ-cell type. With this model it is possible to predict and to gain insight into the changes of the DNA content occurring during the normal process of spermatogenesis. The relative proportions of haploid, diploid, S phase and tetraploid germ cells with increasing age of the rats were calculated. Calculated and actual experimental flow cytometry data were compared to test the accuracy of the model, and these show good agreement. The present work demonstrates that single-parameter DNA analysis of testicular cells is primarily a reflection of germ cells in the spermatocyte and spermatid stages of development, and of non-germ cells. The FCM single-parameter DNA analysis of testicular cells is relatively insensitive to changes in the stem cell and spermatogonial stages of germ-cell development.

Animals↗
PubMed1986

Immuno- and enzyme-histo/cytochemical analysis of resin sections and cell suspensions: a comprehensive diagnosis of bone marrow on a single aspiration sample.

A protocol is described in which a single bone marrow aspiration specimen is used to prepare resin sections and cell suspensions. Using this protocol, a full battery of morphological, enzyme-histochemical, immuno-histochemical and cyto-pathological techniques can be applied. This allows a definitive bone marrow diagnosis to be established without resorting to bone marrow biopsy in the majority of patients.

Antigens↗
PubMed1986

Effects of danazol on spermatogenesis in adult rats.

Adult male Wistar rats were treated with Danazol (4 mg/day s.c.) for 52 days. The drug produced a marked, rapid drop in serum testosterone concentrations to very low levels and caused a slower decrease in serum FSH, LH and testis weight. Flow cytometric analysis of testicular cell suspensions showed a decline in the absolute numbers of haploid cells (spermatids), tetraploid cells (mainly pachytene spermatocytes) and of cells in the S-phase of the division cycle, suggesting that Danazol inhibited proliferation of spermatogonia and/or primary spermatocytes. Histological counting of the different types of spermatogonia, however, revealed no significant change in their numbers during Danazol treatment. It is concluded that Danazol inhibited spermatogenesis primarily after the preleptotene stage of primary spermatocytes.

Animals↗
PubMed1986

Effects of a single injection of a new depot formulation of an LH-releasing hormone agonist on spermatogenesis in adult rats.

Adult male Wistar rats were treated with a single injection (500 micrograms s.c.) of a new biodegradable depot formulation of the LH-releasing hormone (LHRH) analogue [D-Ser(But)6]AzGly10-LH-RH (Zoladex; ICI 118,630) to evaluate its potential for inhibiting spermatogenesis. The drug produced a marked (P less than or equal to 0.05) decrease in serum concentrations of FSH, LH and testosterone with a maximum effect 14 days after treatment. Since striking focal histological changes were seen in the testis after only 1 week, at a time when changes in serum gonadotrophins were minimal, there may be a direct effect of the LHRH analogue on spermatogenesis. Degenerative changes in germ cells as well as Sertoli cells could be observed. Flow-cytometric analysis of testicular cell suspensions showed a significant decline in the absolute numbers of haploid cells (spermatids), tetraploid cells (mainly pachytene spermatocytes) and of the numbers of cells in the S-phase of the cell cycle. This suggests that the drug also inhibits proliferation of spermatogonia and/or primary spermatocytes. Testis weight, serum hormone concentrations, and histological and cytological parameters returned to essentially normal values 52 days after the injection. It is concluded that this new method of administration may have practical and pharmacokinetic advantages for the purpose of reversible inhibition of spermatogenesis.

Animals↗
PubMed1986

Automated deposition of cellular material on glass slides: Cytopress.

The present paper describes an automated apparatus called Cytopress that collects cells or cellular material e.g. nuclei out of suspension on a membrane film by filtration. In a second step this material is transferred on to a glass slide by means of standardized pressure-fixation. Thus slides of high quality are made rapidly and easily that are suited for a variety of stainings.

Adenocarcinoma↗
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

Flow cytometric analysis and sorting of human endometrial cells after immunocytochemical labeling for cytokeratin using a monoclonal antibody.

Endometrial cells in suspension were stained with propidium iodide and a monoclonal antibody against a cytokeratin intermediate filament protein specific for glandular and columnar cells (RGE 53). In this way columnar epithelial cells of the normal endometrium and of adenocarcinomas can be distinguished and separated by flow cytometry from non-epithelial cells (fibroblasts and inflammatory cells) and squamous epithelial cells, all of which are negative for RGE 53. This makes it possible to analyse and also sort pure fractions of this particular tissue type for further studies. The use of propidium iodide allows simultaneous DNA flow cytometry of these columnar epithelial cells. Therefore, the use of antibodies to cytokeratin in combination with propidium iodide can be of help in analyzing and sorting pure fractions of both normal and malignant cells. This allows a more refined examination of complex cell mixtures using flow cytometry.

Adenocarcinoma↗
PubMed1985

DNA analysis and sorting of rat testis cells using two-parameter flow cytometry.

By use of two-parameter flow cytometry of rat testis cell suspensions stained with mithramycin for DNA (the peak amplitude of the fluorescence signal versus total fluorescence intensity integrated over time), eight cell compartments could be distinguished without pre-enrichment of the samples. Cells in these compartments were identified by sorting and subsequent microscopic examination.

Animals↗
PubMed1985

Repeated urine cell culture in soft agar: potential role in follow-up of patients with transitional cell carcinoma.

To evaluate the persistence of cells with clonogenic properties in patients being treated for transitional cell carcinoma, 272 urine samples were collected from 75 patients and cultured in a double layer soft agar "cloning" system. The development of colonies was evaluated with growth curves based on repeated colony counting with an Omnicon automated colony counter at regular time intervals. Forty-eight patients had at least one evaluable culture. Comparing the results of colony development in culture with the clinical evaluation of the patients, 9 patients had a histologically proven recurrence preceded or accompanied by tumor colony growth in urine culture. One patient had tumor recurrence with growth negative urine culture (false negative). Fifteen patients have had growth negative urine culture with a negative follow-up (mean 19.5 months). Twenty-one patients have had growth positive urine cell cultures with no recurrence in their follow-up (mean 18.7 months). Although the follow-up times are at present relatively short, the present study suggests that repeated soft agar urine culture of patients with low-grade, low-stage bladder carcinoma may provide a means for identifying those patients at a higher risk for recurrence/progression of their disease.

Agar↗
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↗
PubMed1985

Squamous and transitional elements in rat bladder carcinomas induced by N-butyl-N-4-hydroxybutyl-nitrosamine (BBN). A study of cytokeratin expression.

Three hundred rat bladders bearing tumors induced by N-butyl-N-4-(OH)butyl-nitrosamine (BBN) were examined by routine histologic study and immunohistochemical staining of intermediate filament types. Smaller lesions were similar to human urothelial dysplasia histologically and immunohistochemically. Progression of the lesions demonstrated large exophytic papillomas with extensive endophytic epithelial growth into abundant stroma. These lesions showed increasing predominance of squamous over transitional elements. Immunohistochemical findings confirmed these results and also demonstrated that morphologically indifferent cells, even in early lesions, express heavier cytokeratins characteristic of keratinizing squamous epithelium. These results demonstrate that BBN-induced bladder tumors show marked quantitative and qualitative differences from the most common, purely transitional, human bladder carcinomas. However, the development in BBN-treated rat bladders of two tumor types, squamous and transitional, from an altered urothelium may serve as an attractive model for further study of the molecular genetics of keratin expression.

Animals↗
PubMed1984

A new disaggregation device for cytology specimens.

A new means of disaggregating cytology specimens in suspension using an immersible rotor device is described. The new rotor is compared to an automated syringing apparatus using cervical samples. Similar results using both devices are obtained for both normal and abnormal specimens.

Cell Separation↗