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Biomedical subjects

C Herman

Publications and source records attributed to C Herman.

54 records · Page 3Linked to original sources

Cytokeratin expression during neoplastic progression of human transitional cell carcinomas as detected by a monoclonal and a polyclonal antibody.

Antibodies to intermediate filament proteins were used to study different cell layers in normal human transitional epithelium, 16 human transitional cell carcinomas, and two cell lines derived from human bladder carcinomas. Conventional rabbit antisera to human skin keratins stained all layers of the transitional epithelium from bladder, ureter, and kidney. A slightly higher staining intensity was found in the basal and superficial layers as compared with the intermediate cell layers. A monoclonal antibody to cytokeratin 18 (RGE 53), however, stained only the superficial cell layer of transitional epithelium, the so-called umbrella cells. In well-differentiated (grade I) transitional cell carcinomas, RGE 53 stained only the superficial cells of papillary structures. In higher grade papillary tumors, RGE 53 also stained cells within the basal and intermediate layers, whereas in high-grade, invasive tumors almost all tumor cells were RGE 53 positive. These results show that monoclonal antibodies to cytokeratins can provide both an indication of processes involved in neoplastic progression of bladder tumors and a means of studying the molecular relationship of the tumor cells to normal cells.

Animals↗

Continuous measurement and analysis of staining kinetics by flow cytometry.

The measurement of color development with time in cells following the start of a staining reaction is of interest in a number of biological systems. These include the subsets of peripheral white blood cells after acridine orange staining, the uptake by cells and nuclei of fluorescent agents, especially antitumor drugs, and measurement of intracellular enzyme kinetics using fluorogenic or absorbing substrates. The present work describes a simple computer program for analyzing flow cytometric (FCM) data versus time, including both the population kinetics of color development and the variability of staining speed within one population of cells. A single-channel absorption measurement in flow (Technicon Hemalog D) was used to record peroxidase kinetics in peripheral blood cells. Every 5 s, a 64-channel absorption histogram was recorded, up to a maximum of 64 histograms. The data were then analyzed by a computer program which searched for the peak channel of each histogram. A least-squares fit was computed for these maxima. The asymmetries of the 64 absorption histograms were compared to see if there was more than one population present with different time constants. Although developed for enzyme kinetic measurements, this program may have wider usefulness in any measurements of time-dependent phenomena by FCM.

Computers↗

Monoclonal antibody to keratin filaments, specific for glandular epithelia and their tumors. Use in surgical pathology.

A monoclonal antibody (RGE 53) against keratin intermediate filaments was prepared by fusing myeloma cells with splenic lymphocytes from mice immunized with a HeLa cell cytoskeleton preparation and human callus keratins. The antibody, selected for fibrillar staining in HeLa cells and a negative reaction on human skin, was tested on frozen sections from normal and malignant human tissues. RGE 53 specifically recognizes glandular epithelial cells from female breast, digestive, respiratory, and urogenital tracts, endocrine and exocrine tissues, and mesothelial cells. No reaction is found in stratified squamous epithelia or nonepithelial tissues. Furthermore, RGE 53 can distinguish adenocarcinomas and mesotheliomas, which stain positively in the indirect immunofluorescence technique from squamous cell carcinomas and nonepithelial tumors, which are negative for this antibody. RGE 53 may, therefore, be a useful tool for differential diagnosis in surgical pathology, especially in those cases in which anaplastic carcinomas have to be characterized.

Adenocarcinoma↗

Quantitative analysis of flow microfluorometric data for screening gynecologic cytology specimens.

Gynecologic cytology specimens that included the entire spectrum of cervical cytology classification were stained with a combination of propidium iodide and fluorescein isothiocyanate, then analyzed using a flow microfluorometer to measure nucleic acid and protein content, respectively. Numerous descriptors of the resulting two parameter distribution (nucleic acid versus protein) were defined. These descriptors included assessment of the presence or absence of abnormal cells. They also included measures of the staining intensity and dispersion of the normal squamous cell population and the intensity of inflammatory response in the cell population. Relative percentages of inflammatory and epithelial cells were demonstrated to effect the screening performance of this system only in borderline lesions. Decision tree algorithms allowed optimization of the selected parameters for screening logic of normal-abnormal decisions on a specimen-by-specimen basis. In addition, quantitative definitions of specimen adequacy were determined. Appropriate controls for batch staining of specimens were evaluated. These results of applying pattern recognition techniques to flow microfluorometer multiparameter data demonstrate that considerably more information about cell populations and subpopulations can be extracted than heretofore possible.

Adolescent↗

Measurement of cellular DNA mass by flow microfluorometry with use of a biological internal standard.

Use of a biological standard (chicken erythrocytes) mixed with experimental cell populations allows control of all variables in flow microfluorometric determination of DNA content. These variables include both staining and instrument procedures. In addition, the use of a biological standard allows determination of cellular DNA mass in unperturbed cell populations. DNA mass measured by FMF technique correlates closely with values reported in the literature that used biochemical techniques.

DNA↗

Drug-induced changes in DNA fluorescence intensity detected by flow microfluorometry and their implications for analysis of DNA content distributions.

Chicken erythrocytes, which contain less DNA than mammalian diploid cells, were used as an internal standard to control instrumental and staining variables during flow microfluorometric analysis. With the DNA stain, mithramycin, and with an EPICS II flow microfluorometer, ratios between the modal G1 fluorescence of experimental cells and that of chicken erythrocytes were determined. The results indicate that unperturbed cell populations of L1210 and HeLa cells in vitro and L1210 ascites cells in vivo have relatively stable fluorescence ratios, although there is a significant difference between the ratios of one L1210 cell line in vitro and another in vivo. In contrast, L1210 ascites treated in vivo with different schedules of cyclophosphamide and Adriamycin showed wide fluctuations in the fluorescence intensity ratios for 96 hr after treatment. Also, differences in the fluorescence ratios were observed between less advanced and more advanced L1210 ascites after treatment with the same schedule. These effects indicate an alteration in DNA staining with mithramycin, brought about by drug treatment that could seriously affect the interpretation of DNA histogram data. Nevertheless, changes in mithramycin staining may prove to be a very important probe to detect persistent drug effects.

Animals↗

Long-term potentiation in vivo in layers II/III of rat barrel cortex.

Long-term potentiation was studied in vivo in the rat barrel cortex. It was found that LTP lasting several hours could be induced in layer II/III by tetanic stimuli applied in layer IV. The probability of inducing LTP at a given site was high (86%) provided that the electrodes were not displaced too far horizontally. LTP was not observed if the stimulating electrode was located on the far side of the neighbouring barrel-column from the recording electrode. The strongest LTP was induced by stimulating layer IV septal locations or the edge of the barrel and recording in the near half of the neighbouring barrel. However, examples were found of LTP from layer IV to II/III within the same barrel, within the same septum and from barrel to adjacent septum. The probability of inducing LTP on a particular occasion was greatly increased by iontophoresis of bicuculline at the recording site during the tetanus (from 20 to 55% judged by a change in peak amplitude). The average increase in the peak amplitude was 29 +/- 3.2% for protocol 1 (urethane anesthesia, monopolar stimulation) and 23 +/- 7% for protocol 2 (barbiturate anesthesia, bipolar stimulation). The probability of inducing LTP was greater if the first tetanus was accompanied by BMI application (67%) than for any subsequent attempts (39%). These results suggest it should be possible to study the effect of LTP on sensory processing in defined positions within the barrel field.

Animals↗

Death personification in the Kevorkian era.

Have death personifications changed over the past quarter of a century? A 1971 study explored the death personifications of college students, student nurses, nurses, and mortuary science students. The present study focused on 123 students of nursing and 324 students enrolled in general study courses at a large university in the American Southwest. The study also explored the possibility that Dr. Jack Kevorkian may have emerged as a contemporary image of death. Respondents were asked to imagine death as a character in the movies and to indicate the nature of this character through fixed-choice alternatives. Respondents were also asked to identify the sources or models for their personifications of death. It was found that male personifications of death still predominate, but that female personifications occur with greater frequency than in the earlier study, especially among female respondents. Although the "Gentle Comforter" is most often seen as an elderly person, overall the relationship between age and death personification appears to have weakened. There is now a larger divergence between the sexes in type of death personification selected most frequently. Females most often select "kind and gentle" imagery; males now are more likely to see death as a "cold, remote" person. Grim and terrifying images of death occur with some frequency, but are the least often selected by respondents of either sex. There was no evidence to support the emergence of a Kevorkian-type death personification, although he was identified correctly by almost all respondents and was viewed favorably by a majority.

Adult↗