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Biomedical subjects

E Greenebaum

Publications and source records attributed to E Greenebaum.

At least 37 records · Page 2Linked to original sources

DNA ploidy of spindle cell soft-tissue tumors and its relationship to histology and clinical outcome.

With the use of fresh cell suspensions, the DNA ploidy of 11 benign and 27 malignant spindle cell soft-tissue lesions was determined by flow cytometry (37 cases) and/or image cytometry (35 cases). Of the 27 malignant lesions, 10 were low- or intermediate-grade sarcomas, and 17 were high-grade sarcomas. In the malignant lesions, the DNA ploidy was correlated with the histologic grade and the clinical outcome. Of the 11 benign lesions, six had a diploid DNA ploidy pattern, and five were nondiploid by either flow cytometry or image cytometry. All benign cases had a favorable outocome regardless of ploidy. Eight of the 10 low- or intermediate-grade malignant lesions were diploid, whereas two were nondiploid. Of the 17 high-grade sarcomas, 15 were nondiploid by either method of measurement and nine by both. Of the 10 diploid malignant tumors, only one had an unfavorable outcome (malignant mesothelioma with biopsy only), whereas of the 17 malignant lesions that were nondiploid, five had no evidence of recurrent disease, three cases recurred, eight patients died of disease, and one patient died of an unrelated cause. We concluded that (1) ploidy does not differentiate benign from malignant spindle cell soft-tissue tumors, (2) the nondiploid DNA pattern is more common in high-grade than in low- or intermediate-grade sarcomas, and (3) there is a statistically significant relationship between nondiploidy and a worse clinical outcome in sarcomas.

Adult↗

Deoxyribonucleic acid ploidy and cell cycle events in benign colonic epithelium peripheral to carcinoma.

DNA ploidy and cell cycle phases of benign human colonic epithelium peripheral to adenocarcinoma were analyzed by flow cytometry in 188 prospective cases. Human colonic epithelium was shown to be diploid with a mean DNA index (DI) of 1.01. The G0G1 compartment accounted for nearly 93% of the cells with the remainder in the S and G2+M phases. Parallel [3H]thymidine uptake on selected cases confirmed the relatively low proliferative activity of colonic mucosa. The DNA index and the cell cycle compartments exhibited no correlation to the ploidy, Dukes' stage, size, and anatomical location of the corresponding malignant tumors. Approximately 25% of the benign samples possessed DI values outside of the diploid range (defined as the mean +/- sd). Analysis of these apparently hypo- (less than 0.92) and hyper- (greater than 1.09) diploid, histologically normal samples in terms of cell cycle kinetics and their relationship to Dukes' stage, location, distance, and the ploidy of the tumor showed no correlation. The only characteristic differentiating these "aberrant" samples from diploid benign tissue was variation in DI possibly due to differences in fluorochrome binding or accessibility to DNA. Whereas these results indicate some degree of variability in the DNA content of benign colonic epithelia, neither DI nor cell cycle kinetics appear to be affected by the presence of a malignant tumor and are not representative of either the ploidy or pathologic stage of the corresponding colonic carcinoma.

Adenocarcinoma↗

Flow cytometry and Feulgen cytophotometry in evaluation of effusions.

Fifty-eight effusions (42 pleural and 16 ascitic fluids) from patients with and without cancer were analyzed by conventional cytology and the results compared with DNA patterns generated by flow cytometry of 10(4) nuclei and several modes of Feulgen cytophotometry. In 31 patients (24 without evidence of cancer and seven with history of cancer and cytologically negative fluids), the fluids were diploid by flow cytometry. One fluid with atypical cells from a lymphoma suspect was also diploid. Flow cytometry of 26 cytologically cancerous fluids disclosed aneuploid DNA patterns in 16 and diploid patterns in ten. Feulgen cytophotometry of 11 of these fluids (three aneuploid, eight diploid) was performed on nuclear preparations identical to those used in flow cytometry and on restrained smears used for visual analysis. The analysis was performed in two modes: as a study of 500 sequential nuclei in an automated system, mimicking flow cytometry, and visually selected large, presumably malignant nuclei. In nine of the 11 cases, the DNA content of visually selected cancer cells was aneuploid, even though this DNA pattern was not evident in the analysis of 500 sequential cells. In two cases, both diploid by flow cytometry, the Feulgen analysis confirmed the presence of cancer cells in the diploid range. In samples of 10(4) nuclei representing a mixed population of cells occurring in effusions, the presence of aneuploid cancer cells may not be disclosed by conventional flow cytometry. A larger sample of cells, a detailed analysis of DNA histograms, and perhaps sorting of select cells in the hypertetraploid range, may prove essential before flow cytometry can be accepted as a diagnostic tool in the laboratory in the assessment of effusions.

Aneuploidy↗

The diagnostic value of flow cytometric DNA measurements in follicular tumors of the thyroid gland.

Flow cytometric analysis of DNA content was performed on single or multiple samples from 34 thyroidectomy specimens. There were 29 thyroids with diploid DNA content, comprising 15 nonneoplastic lesions, 5 follicular adenomas, 1 medullary carcinoma, and 8 papillary carcinomas. Aneuploid DNA pattern was observed in five cases, including one metastatic mammary carcinoma. The initial histologic diagnoses in the remaining four aneuploid thyroid lesions were follicular carcinoma in one and follicular adenoma in three. The abnormal DNA pattern in the three follicular "adenomas" prompted a review of their aspiration cytologic and histologic features. The fine-needle aspiration biopsy was performed in two of the three cases and showed evidence of a follicular neoplasm with significant nuclear atypia in both. Histologic review of the three lesions led to a modified diagnosis of noninvasive low grade follicular carcinoma in all three. Flow cytometric analysis of DNA content may prove to be a highly useful adjunct in the evaluation of follicular thyroid tumors. Long-term clinical follow-up is warranted to document the clinical significance of these observations.

Adenocarcinoma↗

Comparison of needle aspiration and solid biopsy technics in the flow cytometric study of DNA distributions of surgically resected tumors.

Flow cytometric analysis (FCM) of DNA content of nuclei was performed on simultaneously obtained tissue samples and needle aspirates from 37 primary colorectal cancers and from 21 other tumors. There was a marked increase in the proportion of the nondiploid cell population in 18 of 58 aspirates when compared with the corresponding tissue samples, presumably because of selective aspiration of tumor cells. The difference was significant at level alpha much less than 0.01 by a paired t-test and was most pronounced in tumors wherein a nondiploid population constituted more than 20% of the sample. The difference did not correlate with the grade or stage of the tumor. These observations suggest that the sampling of surgically resected tissue specimens for DNA analysis by FCM is performed best by needle aspiration, which may increase the yield of nondiploid cells, does not interfere with histologic diagnosis, and may prove especially useful in the analysis of small surgical specimens.

Biopsy↗

Binding of phorbol dibutyrate and epidermal growth factor to cultured human epidermal cells.

Primary cell cultures of normal human epidermal keratinocytes and melanocytes and human cell lines established from a primary melanoma (SK-PM-4) and metastatic melanomas (HO#1, SK-MEL21, and SK-MEL37) contain specific and saturable receptors for the tumor promoter phorbol dibutyrate (PDBu). Scatchard analyses of the keratinocytes revealed two classes of binding sites: 1) a high-affinity class (affinity constant = 37 nM; 1.3 X 10(6) sites/cell) and a low-affinity class (affinity constant = 4,880 nM; 7 X 10(7) sites/cell). The melanoma cultures, likewise, showed high- and low-affinity classes of PDBu binding sites. However, the affinity constant values and total numbers of sites in the melanoma cells were lower than the corresponding values in the keratinocytes. The binding of [3H]PDBu to human keratinocytes was inhibited by the tumor promoters 12-O-tetradecanoylphorbol 13-acetate and teleocidin but not by phorbol, which lacks tumor-promoting activity. Human serum also inhibited binding. Specific receptors for epidermal growth factor (EGF) were demonstrated in the keratinocytes and primary melanoma cultures. In contrast, three metastatic melanoma cultures gave negligible levels of EGF binding. Among the various cell types, the extent of [3H]PDBu binding did not correlate with the extent of EGF binding, indicating that these two substances occupy distinctly separate types of receptors.

Caenorhabditis elegans Proteins↗

Inhibition of phorbol ester-receptor binding by a factor from human serum.

The inhibition of receptor binding of [3H]phorbol-12,13-dibutyrate (PDBu) by a factor from human serum was characterized. The serum factor inhibited [3H]PDBu binding in intact monolayer cultures of the rat embryo cell line CREF N and in a subcellular system containing membranes from these cells. Inhibition occurred at both 37 and 4 degrees C and was rapid and reversible. An analysis of [3H]PDBu binding in the presence of the serum factor indicated that inhibition of [3H]PDBu binding by the serum factor was noncompetitive. Using gel filtration to separate the serum factor from free [3H]PDBu, we obtained evidence that the serum factor does not act by binding or trapping the [3H]PDBu. Unlike the phorbol ester tumor promoters, the serum factor alone did not stimulate the release of choline or arachidonic acid from cellular phospholipids, nor did it inhibit the binding of 125I-labeled epidermal growth factor to cellular receptors. The factor did, however, antagonize the inhibition of epidermal growth factor binding induced by PDBu. Sera from pregnant women were, in general, more inhibitory of [3H]PDBu binding than were those from nonpregnant women, which were more inhibitory than those from men. During these studies we found that CREF N cells responded to being grown in the presence of PDBu by partial down regulation of the phorboid receptor. The 50% effective dose for down regulation was 8 nM PDBu, and the maximum effect occurred after 6 h. Taken together, our results indicate that the serum factor inhibits [3H]PDBu binding by a direct physical effect at the level of the phorboid receptors or their associated membranes. It would appear that if this factor acts in vivo, then it might antagonize certain effects of this class of tumor promoters.

Animals↗

Identification of receptors for phorbol ester tumor promoters in intact mammalian cells and of an inhibitor of receptor binding in biologic fluids.

Utilizing [3H]phorbol dibutyrate [P(Bu)2], we have developed an assay for high-affinity phorbol ester receptors in intact rat embryo fibroblasts. At 37 degrees C, binding of [3H]P(Bu)2 reached a maximum within 10 min and was rapidly reversible. The tumor promoters 12-O-tetradecanoyl-phorbol 13-acetate, teleocidin B, and mezerein were potent inhibitors of [3H]P(Bu)2 binding. Phorbol and 4-alpha-phorbol didecanoate, which lack tumor-promoting activity, did not inhibit [3H]P(Bu)2 binding. Epidermal growth factor, platelet-derived growth factor, fibroblast growth factor, arginine and lysine vasopressin, luteinizing-hormone releasing hormone, and diazepam did not inhibit [3H]P(Bu)2 binding. A Scatchard analysis was compatible with two classes of binding sites, one with Kd = 8 nM and about 1--2 x 10(5) sites per cell and the other with Kd = 710 nM and about 3 x 10(6) sites per cell. Sera from various species, human amniotic fluid, and certain tissue extracts inhibited specific binding of [3H]P(Bu)2. Fractionation of human serum led to 135-fold purification of an inhibitory factor with a molecular weight in the range 40,000 to 80,000.

Animals↗

Exencephaly-anencephaly sequence: proof by ultrasound imaging and amniotic fluid cytology.

We present and discuss major current theories about the developmental natural history of the anencephalic human fetus. We confirm previous observations made using transvaginal ultrasonography of exencephalic fetuses which were later imaged and/or delivered as anencephalic fetuses. We explore the possibility of proving the theory of the slowly rubbed-off exposed brain tissue by cytologic examination and special staining of aspirated cells in amniotic fluid. Three fetuses with a typical sonographic picture of exencephaly at 13-15 postmenstrual weeks underwent amniocentesis. The aspirated fluid contained pathognomonic neural cells. The same fetuses later showed the characteristic sonographic and postabortion picture of anencephaly. Our results support the theory that exencephaly is the forerunner of anencephaly.

Amniocentesis↗

CA-125 and carcinoembryonic antigen assay vs. cytodiagnostic experience in the classification of benign ovarian cysts.

OBJECTIVE: To compare the relative strengths of two factors involved in making an accurate differentiation between functional and epithelial ovarian cysts, along with their combination: (1) the cytologist's level of experience in interpreting ovarian cytology, (2) the use of the tumor markers carcinoembryonic antigen (CEA) and CA-125 in cyst fluid, and (3) a combination of (1) and (2). STUDY DESIGN: Papanicolaou-stained sediments from fluid aspirated from 31 resected ovarian cysts (6 functional and 25 epithelial) were blindly and independently evaluated by five pathologists with varying experience in ovarian cytology. Cyst fluid supernatant was used for CEA, enzyme-linked immunosorbent assay, and CA-125 radioimmunoassay; CEA levels > 5 ng/mL or CA-125 > 5,000 U/mL were considered elevated. Cysts were categorized cytologically and histologically as functional or epithelial and by tumor markers as "neither elevated" or "either or both elevated" (EBE). RESULTS: The agreement of histologic diagnosis with each pathologist's cytologic diagnosis ranged from 53% to 84% (53%, 71%, 83%, 82%, 84%), corresponding to increasing level of experience. The percentage of agreement with EBE was 77%, whereas combined experienced pathologist's diagnosis and EBE was 87%. Kappa equaled .45 for experienced cytopathologist's diagnosis or EBE alone. Kappa equaled .53 when the pathologist or EBE diagnosed an epithelial cyst, indicating results unlikely to occur by chance. CONCLUSION: The distinction of functional from epithelial ovarian cysts is best achieved by combining measurement of the tumor markers CEA and CA-125 with a high level of cytopathology experience.

Adult↗

Aspiration cytology of ovarian cysts in in vitro fertilization patients.

Transvaginal ultrasonography is routinely performed in the course of egg retrieval in patients presenting for in vitro fertilization (IVF) and results in the discovery of occult, subclinical ovarian cysts that would otherwise have gone undetected. This study (1) evaluated the cellular composition of the cyst fluids based on a comparison with cells obtained from cysts of known and documented histologic type, and (2) attempted to determine if the cytologic evaluation of cyst fluid was necessary to exclude occult ovarian cancer. During the 1.8 years of our study, 931 patients underwent 1,544 ultrasound-guided ovum retrievals; during this same period, 90 specimens of ovarian cyst fluid were examined. Of them, none contained cancer cells. A single case of cystic ovarian cancer was detected by ultrasound but was not aspirated. The cytologic diagnosis of endometriosis was established in 12 specimens from 10 patients, 5 of whom did not have a previously documented clinical diagnosis of endometriosis. The role of routine ovarian cyst fluid cytology as part of an IVF protocol may be of limited value in cancer diagnosis. However, until the incidence of ovarian cancer in the subset of women with both infertility and ovarian cysts is known, it would seem prudent to continue to examine any voluminous or discolored ovarian cyst fluid obtained from IVF patients. The presence of ovarian cysts did not affect the clinical pregnancy rate per retrieval.

Adult↗

Cytologic manifestation of an unusual bacterial form, Simonsiella species.

Puzzling rodlike structures overlying benign squamous cells in exfoliative cytologic specimens from the upper gastrointestinal and respiratory tracts were initially considered to be fungi, protozoa, bronchodilator crystals, hemoglobin tactoids or plastic fragments. Their morphologic similarity to Simonsiella, a gram-negative bacteria frequently found in the oral cavity, was ultimately recognized. Further studies of smears and cultures obtained from the oral cavities of the authors and from the American Type Culture Collection confirmed the nature of the original findings. These giant bacterial forms were usually found in caterpillarlike side-by-side arrangements of 10 to 12 organisms. Cytologists should be aware of their appearance to avoid possible confusion with pathogenic organisms.

Bacteroidetes↗