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Biomedical subjects

C J Herman

Publications and source records attributed to C J Herman.

At least 19 recordsLinked to original sources

Cytometric DNA analysis in the management of cancer. Clinical and laboratory considerations.

Clinically, cytometric DNA (ploidy and proliferative fraction) analysis is primarily used in the determination of tumor prognosis. This determination has a clinical impact primarily in low stage tumors where therapeutic options are available, rather than in higher stage tumors that will be treated the same independent of their ploidy or proliferative fraction. In general, the best available data indicate that for most tumors aneuploidy is an indicator of more aggressiveness. Ploidy analysis is independent of other prognostic indicators in many tumors and is the most significant prognostic indicator in some. More recent data suggest that proliferative fraction, the percentage of tumor cells in the S-phase of the cell cycle, may be as important an indicator of tumor prognosis as simple presence of aneuploidy. However, the poor interlaboratory reproducibility of S-phase measurements as currently performed limits the general applicability of this measurement in the management of patients.

Cell Division

Flow cytometric DNA content of fresh tumor specimens using keratin-antibody as second stain for two-parameter analysis.

Studies concerning flow cytometric assessed DNA content reveal problems in interpretating DNA histograms of tumor specimens. The main problems are histograms with a broad coefficient of variation in the G0/G1 fraction; a high G2M fraction and samples with a low percentage of tumor cells. Therefore, in the present study, 382 fresh tumor specimens of carcinomas were analysed routinely, double labeled with, on the one hand, propidium-iodide for assessing DNA content and, on the other, a monoclonal keratin-antibody for marking epithelial and tumor cells. Of the 311 tumor samples, using single parameter analysis 165 (54%) were classified as DNA aneuploid and 146 (46%) as DNA "euploid." By double parameter analysis, 224 (72%) samples were keratin positive and 87 (27%) keratin negative and, of the 224 keratin positive tumors, 175 (78%) were DNA aneuploid and 49 (22%) DNA euploid. The DNA histograms of single and double parameter analysis were compared and it was concluded that in 24 cases (11%) keratin labeling was necessary to recognize DNA aneuploidy. In another 23 (10%) cases, keratin labeling was helpful in assessing DNA aneuploidy. Finally when the results of the 311 samples were combined, 215 (68%) were scored as DNA aneuploid and 99 (32%) DNA euploid. Thus the overall gain in assessing DNA aneuploidy using the double labeling technique is 14%. In conclusion, it is shown that keratin labeling on fresh tumor cell suspensions of epithelial tumors is of additional value in establishing DNA content. Because single parameter DNA assessment is adequate in approximately 60% of the tested samples, the double labeling technique can be performed routinely, or after initial single parameter DNA assessment. Histograms having a broad CV and/or a high G2M are good candidates for the double labeling technique. Using this technique, DNA-content assessment becomes more reliable.

Antibodies

Hydrolysis kinetics of phospholipids in thermally stressed intravenous lipid emulsion formulations.

A model 20% w/v emulsion, prepared with either a commercially available pharmaceutical grade soy oil or a highly purified grade of oil from the same origin and stabilized with a commercially available mixture of egg yolk phospholipids was passed through a Microfluidics homogenizer until the mean particle size fell below 500 nm diameter. Samples stored in sealed all-glass ampoules were thermally stressed over a temperature range of 5-90 degrees C and samples taken at appropriate intervals for analysis by HPLC. Hydrolysis degradation kinetics were in conformation with the Arrhenius equation. The energy of activation for phosphatidylcholine was virtually identical for emulsions prepared with either pharmaceutical or purified oil (65 and 63 kJ mol-1, respectively). For phosphatidylethanolamine itself the respective activation energies were 53 and 54 kJ mol-1, suggesting that the source of the oil used in preparing the emulsions had no significance in the degradation processes of the resulting systems.

Chromatography, High Pressure Liquid

Reproducibility of FCM DNA content from replicate paraffin block samples.

Fifty-five paraffin blocks of morphologically and clinically normal colon specimens were divided in half and the two halves treated as separate aliquots for determination of DNA content using propidium iodide staining and FCM analysis. Forty-five of the 55 samples showed a peak channel difference between the two aliquots from the same block of greater than 1 channel. Seven of the 55 specimens showed peak channel difference greater than or equal to 10 channels. Similar results were seen if the mean channel of the G0G1 peak was used in place of the peak channel, with 5 of the 55 specimens showing mean channel differences of greater than or equal to 10 channels. The seven samples with peak channel difference greater than or equal to 10 channels were reprocessed and five of the seven showed reduced peak channel difference on reanalysis. The mean peak channel difference of the 55 samples was 4.3 +/- 10.5 (mean +/- 2 S.D.) using the first analysis. These data suggest that caution should be used in interpreting ploidy of nuclei extracted from paraffin block material if the interpretation is based on comparing two aliquots from the same block, even if they are processed and analyzed in the same batch.

Cell Nucleus

DNA content of Hurthle cell nodules in autoimmune thyroiditis.

Hurthle cells are found in thyroid neoplasms and in reactive nodules in thyroiditis or goitrogenic processes. Cytometric studies have evaluated Hurthle cell neoplasms but not their reactive counterparts. DNA content of Hurthle cells in 22 cases of autoimmune thyroiditis was measured by flow cytometry and image content of Hurthle cells in 22 cases of autoimmune thyroiditis was measured by flow cytometry and image processing using nuclei extracted from paraffin-embedded tissue after microdissection of the Hurthle cell nodules. All 22 autoimmune thyroiditis Hurthle cell nodules were diploid, including 16 without associated neoplasms and six with associated malignant neoplasms (four papillary carcinomas, one follicular carcinoma and one follicular adenoma with papillary carcinoma). Concordance between flow cytometry and image processing was 100%. These findings indicate that the markedly atypical Hurthle cells in autoimmune thyroiditis are diploid by DNA quantitation. This suggests that atypia in Hurthle cells due to reactive or neoplastic processes may be differentiated by quantitative DNA analysis.

Adult

The Multivariate Prognostic Index and nuclear DNA content are independent prognostic factors in primary breast cancer patients.

The predictive value of a previously described Multivariate Prognostic Index (which incorporates weighted values of the mitotic activity index, tumor size, and the axillary lymph node status), and the nuclear DNA content (DNA) was evaluated in 156 patients with primary invasive ductal breast cancer, diagnosed between 1980 and 1983. The results were analysed with respect to the occurrence of distant recurrence and survival of the patients after at least 3 yr of follow-up (range 36-73 months; median 44 months). Known prognostic factors such as lymph node status, tumor size, and the mitotic activity index correlated independently with distant recurrence. Furthermore, in respect to survival, the investigated prognostic factors (except DNA content) were significantly correlated. The results indicate that the predictive value of the Multivariate Prognostic Index (MPI) is stronger (P less than 0.001) than of the nuclear DNA content (P less than 0.005) with respect to distant recurrence. In a Cox multivariate regression analysis DNA ploidy turned out to be an independent prognostic factor once the MPI was selected. Furthermore, in Cox's analysis, DNA ploidy was the fourth selected variable after lymph node status, mitotic activity index, and tumor size in individual parameter analysis. The results of this study indicate that, with respect to breast cancer screening programs, it seems worthwhile to integrate morphometric features, the MPI, and DNA ploidy in a new prognostic model.

Adult

The predictive value of repeated urine cell culture in soft agar for recurrences of transitional cell carcinoma.

To evaluate the predictive value for recurrences of repeated urine cell culture in double layer soft-agar in patients with transitional cell carcinoma, 272 urine samples were collected from 75 patients. After a limited period of follow-up (mean 19.5 +/- 15.1 months) nine out of 46 patients with at least one evaluable culture and evaluable for clinical follow-up had a histologically proven recurrence preceded or accompanied by tumor colony growth in culture; one patient had tumor recurrence with growth negative urine cell culture; 15 patients had growth negative urine cell cultures and at first evaluation no recurrence of transitional cell carcinoma; 21 patients had growth positive urine cell cultures and no recurrence at first evaluation of follow-up. These two latter groups were evaluated again after an additional 30 months of follow-up. In the group with growth positive urine cell cultures (21 patients) nine patients (47.6%) developed a recurrence; 10 developed no recurrence. In the other group of patients with growth negative urine cell cultures (15 patients) six patients (40%) developed a recurrence, three following a growth positive urine cell culture. In the group of 46 evaluable patients in this study, 25 patients developed a recurrence, 21 had at least one growth positive culture. The sensitivity is 84% (21/25). Of 33 patients with positive cultures 21 have had a recurrence (64%) while 12/33 (36.7%) are "false positive." The false negative rate is 36.4% (four of 11 patients developed recurrences not preceded by growth positive urine cell cultures).(ABSTRACT TRUNCATED AT 250 WORDS)

Agar

Cytology, immunopathology and flow cytometry in the diagnosis of pleural and peritoneal effusions.

There were 106 pleural and peritoneal effusions studied in order to investigate the contribution of immunocytochemistry and flow cytometry to routine cytologic diagnosis. A panel of antibodies (to cytokeratin, vimentin, human milk fat globule, epithelial membrane antigen and carcinoembryonic antigen) was applied to aceton-fixed slides, using immunoperoxydase and immunofluorescence methods. Flow cytometry was performed using a double labeling method, i.e., propidium iodide for DNA staining and keratin for labeling of epithelial cells. In this way DNA aneuploidy was more evident in the histograms when the fluid contained many reactive nonepithelial cells (lymphocytes). A designation of marker profiles was made for the three most frequently occurring diagnoses, i.e., reactive mesothelial proliferation (I), adenocarcinoma (II), and malignant mesothelioma (III). For the differentiation between adenocarcinoma and malignant mesothelioma, carcinoembryonic antigen was the most useful marker as 75% of the adenocarcinomas was carcinoembryonic antigen-positive and the malignant mesotheliomas were consistently negative. Furthermore, evident DNA-aneuploidy strongly supported the diagnosis of adenocarcinoma, as most malignant mesotheliomas were DNA-euploid, even though occasional DNA-aneuploidy was found in malignant mesotheliomas when different effusions from the same patient were examined. For the differentiation between reactive mesothelial cells and malignant mesothelioma human milk fat globule and/or epithelial membrane antigen, in this study proved to be most reliable, their presence strongly indicating malignancy. It is stressed that the method used (fixation, antibodies, washing procedures) can influence these findings. In 16 patients (17%) performing immunopathology and/or flow cytometry meant an important contribution to diagnosis.

Adenocarcinoma

The relation between mixed mesodermal tumors and adenocarcinomas of the ovary. An immunopathologic study.

Mixed mesodermal tumor of the ovary is a rare neoplasm. The diagnosis is made if an ovarian carcinoma contains heterologous sarcomatous differentiation. With the help of an immunopathologic stain for desmin, the intermediate filament protein of muscle cells, twenty cases of ovarian adenocarcinoma were examined for the presence of morphologically unrecognizable heterologous tumor cells. Six of twenty tumors were positive. There was no significant correlation between a number of clinical parameters and desmin positivity. The authors conclude that mixed mesodermal tumor of the ovary may not be a separate entity. Instead, there may be a single group of ovarian epithelial tumors ranging from borderline or well-differentiated carcinomas to poorly-differentiated tumors which may show homologous (carcinosarcoma) or heterologous (mixed mesodermal tumor) differentiation.

Adenocarcinoma

Survival of spermatogonial stem cells in the rat after split dose irradiation during LH-RH analogue treatment.

A rat model has been created in which a single injection of an LH-RH analogue depot preparation (Zoladex, ICI 118630) produced a temporary interruption of the pituitary-gonadal axis. This effect applied during irradiation was investigated as a possible mechanism to protect the testis from radiation damage. A local testicular irradiation dose of 6.0 Gy was given either as a single dose or as a fractionated (2 X 3.0 Gy) dose at different time intervals ranging from 8 to 72 h. Stem cell survival was measured 11 weeks after irradiation by means of the repopulation index and the number of haploid cells (spermatids) measured by flow cytometry. Serum gonadotrophins and testosterone concentrations were measured to evaluate hormonal recovery. No significant differences were observed between serum concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH) and testosterone and the duration of the fractionation interval. Stem cell survival was higher following fractionated irradiation in comparison with the single dose. For the 8 h interval an increase in recovery ratio was found, amounting to a factor of 5 of the single dose value. The fluctuating pattern of the recovery curves indicated changes in radiosensitivity of stem cells. The combination of hormonal inhibition of spermatogenesis and fractionated irradiation led to a decrease in the absolute numbers of stem cells. However, the stem cell recovery curves were identical to those seen without hormonal inhibition. It was concluded that hormonal pretreatment with Zoladex during split dose irradiation had no protective effect on stem cell survival.

Animals

Renal hamartoma associated with renal cell (Grawitz) tumor: another indication that Grawitz tumors are carcinosarcomas.

We report a case of renal hamartoma (angiofibroma) which contained nests of renal cell (Grawitz) tumor and review the literature regarding this subject. We believe that the finding of Grawitz tumor cells within a mesodermal tumor supports our hypothesis that renal cell (Grawitz) tumors are of mixed (mesodermal and endodermal) origin, that is, they show at least one important biological characteristic of carcinosarcomas.

Carcinoma, Renal Cell

Leukocyte functions.

Knowledge concerning leukocyte functions has increased enormously over the last two decades, largely because techniques have become available to assess them. It has provided insights into many disease entities, as well as on immunologic functioning as a whole. In addition to some classical clinical entities, such as severe combined immunodeficiency disease and chronic granulomatous disease, new entities have been discovered and described, in which subtle disturbances in white blood cell function are sufficient to cause disease. Furthermore, much has been learned about the effect of drugs and disease conditions on leukocyte functions. This review is not intended as a comprehensive description of white blood cell function testing, rather, it is meant to provide a practical guide for clinical laboratories. Therefore only those functional tests are presented which have a direct impact on patient management. These are primarily those functional derangements in white cells which either define a disease entity or without which a diagnosis cannot be made. Granulocytes, monocytes, and lymphocytes will be discussed separately. A short review of their physiology and function is presented to provide a context for the further discussion of their dysfunctions. This discussion of dysfunction and the techniques for laboratory testing thereof includes the evaluation of diagnostic usefulness and implications for patient management. Clinically, the most important granulocyte defects involve chemotaxis, respiratory burst, and intracellular microbial killing. Defects of granulocyte chemotaxis define the lazy leukocyte syndrome, Job's syndrome, and a series of relatively rare idiopathic neutrophil disorders. In addition, chemotaxis is a relatively easily measured sign of actin dysfunction. Defective generation of the respiratory burst with consequent impaired microbial killing defines chronic granulomatous disease and its variants as well as glutathione peroxidase deficiency. The most reliable clinical laboratory testing of granulocyte chemotaxis is performed by using granulocyte migration in response to a chemoattractant, either across a filter or in agarose. For clinical testing of respiratory burst, nitroblue tetrazolium (NBT) reduction is simple, quantitative, and has recently been adapted to automated measurement. Direct measurement of microbial killing is generally not indicated, since clinically important defects are recognized by measurement of respiratory burst generation. Although the monocyte's functions are numerous, they are somewhat less well characterized than those of granulocytes and lymphocytes.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals

Recent progress in clinical quantitative cytology.

The last several years have seen changes in quantitative cytologic procedures that have significantly expanded the clinical research and diagnostic applications of quantitative cytologic analysis. Intact nuclei can be isolated from formalin-fixed paraffin-embedded tissue, and the DNA distribution of these nuclei can be analyzed either with flow cytometry or with slide-based image-processing techniques. In addition, more informative DNA analysis has been achieved using antibodies to bromodeoxyuridine-modified DNA, proliferation-associated nuclear antigens, or tissue-type-specific cytoplasmic antigens as second stains for two-parameter analysis. Improvements in both staining and instrumentation have also increased the usefulness of flow cytometry in analysis of "rare events," ie, cell types comprising less than 0.1% to 0.5% of a total cell suspension. Commercially available slide-based image-processing systems have been developed that provide sophisticated cell and tissue analysis functions using inexpensive personal-computer-based systems.

Antibodies

Cytopress: automated slide preparation of cytologic material from suspension.

This paper describes a new automated system to prepare slides of cytological material from suspension. The system collects material on a filter tape by filtration and transfers it to glass slides by means of pressure-fixation. Using cervical cells as a model, results show that a well-defined cell number is evenly deposited over a standardized area, while a small number of cells is retained on the tape and a negligible number lost in the filtrate. Contamination is very small. Application of the system to other cytological material (fine needle aspirations, monolayer and cell suspension cultures, agar cultures, and isolated nuclei) is shown. In general, more than one slide can be made from one sample. Several histological staining procedures as well as immunofluorescence labeling protocols can be applied to the preparations obtained in this way. This system thus introduces a method that will standardize specimen preparation, is quick, saves operator time, and can be used for both diagnostic and research applications.

Autoanalysis

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

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