Configuration of surfaces of human cancer cells in effusions. A scanning electron microscopic study of microvilli.

The surfaces of viable cells of metastatic human carcinomas of various histologic types and primary origin, suspended in pleural and ascitic fluids, were shown by scanning electron microscopy to be covered by microvilli of variable configuration and distribution. Microvilli of some cancer cells appeared biologically active since they were capable of forming extensions and anastomoses when settling on glass. The possible specificity and significance of microvilli in the light of the experimental data were discussed.     

Choroid plexus carcinoma. Cytologic identification of malignant cells in ascitic fluid

Abdominal deposits of a choroid plexus carcinoma in a patient with a ventriculoperitoneal shunt were cytologically diagnosed by examination of ascitic fluid after regression of the primary tumor. The morphology of the malignant cells in ascitic fluid was more similar to that of mesothelial cells than to the appearance of cells from this lesion in cerebrospinal fluid.     

Comparison of cellular phenotypes in tumor cyst and ascitic fluid from ovarian serous carcinoma.

Density gradient centrifugation was applied to isolate cell subsets from tumor cyst and ascitic fluid in eight patients with ovarian serous carcinoma. A comparison of cellular composition and immunologic reactivity of cells from the cysts and from ascitic fluid in each patient was performed. Some differences in density profiles were found, but in each case the consistency of morphologic cell forms in the primary tumor and ascites was documented. Immunophenotypic analyses of isolated cellular fractions using polyclonal and monoclonal antibodies against ovarian carcinoma-associated antigens showed significant immunologic intratumoral heterogeneity. However, there was a similarity of antigen expression in cells from the primary tumors and ascitic fluids. Our study indicated that morphologic and antigenic characterization of a given tumor could be determined in a single representative sample of ascitic fluid.     

Employment of synchronized cells and flow microfluorometry in investigations on the JB-1 ascites tumour chalones.

In most experimental ascites tumours the growth rate decreases with increasing age and cell number. This decrease is caused by a prolongation of the cell cycle and an increasing accumulation of non-cycling cells in resting (or quiescent) G1 and G2 compartments. In cell-free ascitic fluid from the JB-1 ascites tumour in the plateau phase of growth lowmolecular-weight substances have been found which reversibly and specifically arrest JB-1 cells in G1 and G2. The present paper describes an in-vitro model for testing the effect of the humoral growth inhibitors contained in the ascitic fluid. The test system is based on synchronized JB-1 cells analysed by flow-through cytofluorometry. Addition to the synchronous cells of a ultrafiltrate (less than 50000 Daltons) of the JB-1 ascitic fluid was found to induce a complete, but temporary arrest of the cells at the G1-S border.     

Chalone-like inhibition of Ehrlich ascites cell proliferation in vitro by an ultrafiltrate obtained from the ascitic fluid.

An aqueous ultrafiltrate (10 000-50 000 dalton) prepared from the cell-free ascitic fluid of mice bearing Ehrlich ascites tumour (EAT) in the plateau phase of growth (12-16 days after transplantation) was investigated with regard to its inhibitory effects on the proliferation of EAT cells in a 24-hr suspension culture. The following results were obtained: (1) The in vitro proliferation of cells obtained from the plateau phase of in vivo growth was reversibly inhibited. (2) The dose-response curves show a plateau with a maximum inhibition of about 50%, which suggests that not all cells can be affected. (3) Young cells (4-6 days after transplantation) were not inhibited. (4) Preincubation of plateau phase cells in the culture medium before treatment abolishes the inhibitory effect of the ultrafiltrate. This effect of preincubation is dependent on time and serum concentration. It provides the possibility to differentiate between true "chalone-like" and cytotoxic effects. (5) the inhibitory properties of the ultrafiltrate are destroyed by heating or trypsin treatment. (6) Extracts prepared in the same way from ascitic fluid of mice bearing lymphocytic leukemia L1210 do not inhibit the proliferation of EAT cells. Corresponding extracts from ascitic fluid of mice bearing myelocytic leukemia YM were found to be inhibitory; however, the inhibitory effect was also found on preincubated cells and is therefore considered to be due to an unspecific cytotoxicity. In conclusion, evidence was obtained for a factor from the ascitic fluid of mice bearing EAT, which prevents EAT cells from entering the proliferating state.     

Changing tumour antigen expression in metastatic hepatocellular carcinoma cells of the guinea pig

Hepatocellular carcinoma cells (Line-10), obtained from ascitic fluid after diethylnitrosamine treatment of Sewall Wright strain-2 guinea pigs, produce solid (primary) tumours, lymph-node and lung metastases and malignant ascites when reinjected into animals of the same strain. Monoclonal antibodies were raised against the tumour cells by immunizing BALB/c mice with viable ascitic hepatocellular Line-10 tumour cells. Three hybridomas producing anti-Line-10 monoclonal antibodies were selected for further studies (10TL1, 10TL40 and 10TL43) and compared with monoclonal antibodies against intermediate filament keratins. The anti-Line-10 monoclonal antibodies did not cross react with Line-1 hepatocellular carcinoma cells, nor with normal guinea pig hepatocytes. When ascitic Line-10 cells form high papillary projections on the peritoneal surface, they significantly reduced their antigen expression of 10TL40 and of 10TL43 defined antigens, while the expression of 10TL1 defined antigens remained unaltered. Invading Line-10 cells in the deep submesothelial stromal tissue, however, lost reactivity with MoAb 10TL43 but not with the MoAb's 10TL40 and 10TL1. The antigens on lung- and lymphnode metastases remained largely unaffected. The reactivity with MoAb's 10TL40 and with 10TL43 was also lost upon prolonged culturing of Line-10 cells. The reactivity of Line-10 and Line-1 cells with all monoclonal antibodies against keratin filaments remained unaltered. Line-1 cells could be distinguished from Line-10 cells by the absence of any reactivity with the MoAb's 10TL1, -40, -43, but also by the fact that 100% of the Line-1 cells were positive with antibodies against keratins 5/8, 18, and 7.(ABSTRACT TRUNCATED AT 250 WORDS)     

Gastrointestinal stromal tumor in ascitic fluid. A case report

BACKGROUND: There are few published data on the cytologic features of gastrointestinal stromal tumors (GISTs) in ascitic fluid and whether these features may mimic those of other malignancies. CASE: An 80-year-old woman presented with ascitics associated with multiple intraperitoneal masses. Cytologic examination of the ascitic fluid showed numerous three-dimensional clusters of epithelioid cells. These features and the presence of large, intracytoplasmic vacuoles raised a possible diagnosis of adenocarcinoma. However, mucin could not be demonstrated in the vacuoles, and the cells showed immunoreactivity for vimentin and c-kit but not for cytokeratins. Eighteen months earlier the patient had undergone a partial gastrectomy for a GIST, which predominantly comprised vacuolated, epithelioid cells. The immunoprofile of the primary tumor was identical to that of the ascitic fluid cells. CONCLUSION: GIST cells may closely mimic adenocarcinoma cells in ascitic fluid. Distinguishing between the two neoplasms has important clinical repercussions and is aided by histochemical and immunocytochemical studies--in particular, c-kit immunostaining.     

Tissue-specific inhibition of cellular proliferation in Ehrlich ascites tumor]

The extract of cells of ascitic Ehrlich's tumour (chaloun) and its cell-free fluid produced a marked inhibitory action on the cell proliferation of this tumour four hours after the administration. The effect is tissue-specific, more pronounced in the extract and depends on the dose of the antigen. Eight hours after the extract or the cell-free fluid administration the mitotic activity in the tumour proved to increase in comparison with control; this indicated the presence of a short-lived chaloun action on the G2-phase of the mitotic cycle and synchronization of cell division.     

Changes in cell surface primary cilia and microvilli concurrent with measurements of fluid flow across the rabbit corneal endothelium ex vivo

Primary cilia and microvilli have been reported on the mammalian rabbit corneal endothelium but their relationship to cell function is undefined. Six corneas from healthy 2 kg female albino rabbits were glutaraldehyde-fixed post mortem (15:00 h) or twelve corneal stroma-endothelial preparations incubated at 37 degrees C under an applied hydrostatic pressure of 20 cm H2O for 4 h prior to fixation. The corneal endothelium was assessed by quantitative scanning electron microscopy. Cells fixed immediately post mortem were decorated with small stubby microvilli (average 21 +/- 13/100 micron 2), and only 25% of the cells were decorated with primary cilia having an average length of 2.44 +/- 1.56 microns. Following 4 h ex vivo incubation with a phosphate-buffered Ringer solution, conspicuous microvilli developed to an average density of 40 +/- 19/100 micron 2 and primary cilia were found on 12% of the cells, having on average length of 2.27 +/- 1.38 microns. Following 4 h incubation in a bicarbonate-buffered Ringer solution, small stubby microvilli developed to a density of 49 +/- 18/100 micron 2, and 40% of the cells showed primary cilia with an average length of 4.31 +/- 1.93 microns; the net trans-endothelial fluid flow in the latter set was 60% greater. These studies indicate that the primary cilia on corneal endothelial cells might be responsive to fluid flow, but that mild mechanical and/or chemical stress could also be the cause of the change since the elaboration of primary cilia can be accompanied by microvilli as well.     

A comparison of galactosyltransferase activity in mouse ascites lymphoma cells (Ly/Ya), in cultured lymphoma cells, in ascitic fluid and culture medium

Comparative studies were carried out on the galactosyltransferase activity in ascites lymphoma cells isolated from mouse with ascitic lymphoma Ly/Ya, in these cells grown in vitro (24 hrs culture), in ascitic fluid and culture medium. The effect of varying amounts of UDP-galactose on transfer rate of galactose to ovomucoid by the cell enzyme (ascitic and cultured lymphoma cells) and by the soluble enzyme (ascitic fluid and culture medium) was studied. The activity of the enzyme in the cell culture medium was 2.5-fold higher than that in ascitic fluid. The apparent Km values for UDP-galactose of the enzyme from both kinds of cells and from the two fluids was 7.14 x 10(-7) M. At saturating concentrations of donor substrate, V values for the cells and culture medium was 765 pmoles/10(6) cells/h and 180 pmoles/10(6) cells/h for the ascitic fluid.     

Sulfated glycosaminoglycan and collagen patterns in parietal yolk sac carcinoma (PYSC)

Electrophoretic analyses of collagenous material have shown that the parietal yolk sac carcinoma (PYSC) ascitic tumour synthesizes polypeptide chains that migrate as type IV procollagen. Having molecular weights of 185,000 and 160,000, these polypeptides are sensitive to collagenase. When the PYSC cells are injected subcutaneously, they form a solid tumour, and type I collagen predominates. The electrophoretic analyses of sulfated glycosaminoglycans and enzymatic degradation have shown a predominance of heparan sulfate in the ascitic tumour, and of chondroitin sulfate B in the solid tumour. Cells cultured from ascitic tumours have maintained the same collagen and sulfated glycosaminoglycan patterns as the original cells, whereas in the solid tumour culture only chondroitin sulfate AC has been detected.     

The value of calretinin and cytokeratin 5/6 as markers for mesothelioma in cell block preparations of serous effusions

Objective: To determine the value of calretinin and cytokeratin (CK) 5/6 in discriminating mesothelioma from adenocarcinoma in serous effusion specimens. Methods: A total of 101 recent, histologically or clinically confirmed malignant effusions with immunostained cell block preparations were reviewed. The cases consisted of 34 mesotheliomas and 67 adenocarcinomas. This included 17 ascitic fluid and 84 pleural fluid samples. The adenocarcinomas included metastatic carcinomas from the breast (12), lung (19), stomach (3), colon (1), pancreas (2), ovary (6) endometrium (1) and 23 histologically confirmed metastases from unknown primary sites. The cases were assessed as negative or positive (>5% of cells stained). The staining pattern was recorded as cytoplasmic, cell membrane, nuclear or cytoplasmic and nuclear staining. Results: Calretinin staining was present in 97% (33/34) of the mesothelioma cases with a majority of them showing both cytoplasmic and nuclear staining (29/33). Only 3% (2/67) of adenocarcinomas were positive for calretinin, one being a lung adenocarcinoma and the other an adenocarcinoma of unknown primary site in an ascitic fluid. Cytokeratin 5/6 staining was also present in 33/34 (97%) of mesothelioma cases. Six (9%) adenocarcinomas were positive, including metastases from the lung (1), breast (1), ovary (2) and unknown primary site (2). Four of the six adenocarcinoma cases positive for CK5/6 were in ascitic fluids. No cases of mesothelioma were negative for both calretinin and CK5/6. Only one adenocarcinoma case, (which was from unknown primary site in an ascitic fluid sample), was positive for both markers. Conclusions: The results confirm that calretinin and CK 5/6 are useful markers for mesothelioma in effusion specimens. CK5/6 staining may be less useful for peritoneal fluid specimens where metastatic adenocarcinomas may be more likely to express the antigen. Further study of ascitic/peritoneal specimens is warranted. However, positive staining, particularly for both antigens, is highly indicative of a mesothelial origin for cells. The two markers make a useful addition to EMA and the panel of adenocarcinoma markers routinely applied to effusion specimens.     

Adaptation of Zajdela ascitic hepatoma cells to monolayer growth: change in the cell ganglioside pattern

The Zajdela hepatoma is a transplantable ascitic tumor of the rat, characterized by a very simple ganglioside pattern, GM3 being the main compound. When these cells are adapted to monolayer culture, they undergo a maturation process and the total cellular ganglioside concentration increases progressively; GM2, GM1 and GD3 amounts rose and GD1a accumulated. These modifications in the ganglioside pattern complexity are not affected by the addition of ascitic fluid to the cultures, nor by growth in serum free, hormone-supplemented medium. They are totally reversible when the cultured hepatoma cells are reinjected into a rat and developed an ascitic tumour. Cell growth control and adhesion processes could be related to the maturation process of these hepatoma cells growing in monolayer, which may constitute a convenient model for further investigations on the regulation of membrane glycolipid composition by the external environment.     

Nonspecific suppressive effect of Ehrlich tumor ascitic fluid on transplantation immunity]

The influence of syngeneic Ehrlich ascites tumour fluid on the survival of allogeneic skin allografts was investigated on CC57BR mice. A 3--4-day delay of the allograft rejection in mice injected with ascitic fluid was revealed. Preliminary transplantation of such allograft to mice with Ehrlich tumour produced no intensification of the immunodepressive action of the ascites fluid obtained from them. The data obtained indicated the preferential significance of the antigen-independent immunosuppressive factors of ascites tumour fluid in the suppression of the transplantation immunity in vivo.     

Resistance of ascitic fluid immunosuppressive factor DNA to pancreatic deoxyribonuclease]

It was found that incubation of Ehrlich tumour ascitic fluid with pancreatic DNAse does not result in a destruction of 5S DNA in the immunodepressive factor. After phenol deproteination of the ascitic fluid the resistance of this nucleic factor to DNAse is lost. It is possible that the native nucleic factor is represented by DNP and contains a protein component. During polyacrylamide gel electrophoresis the mobility of the nucleic factor labelled with [3H]-thymidine and isolated after chromatography on hydroxyapatite is found to be nearly the same as that for transferine.     

Studies on the cholinesterase forms present in the ascitic fluid of Ehrlich tumour.

Cholinesterase activity was detected in the ascitic fluid of Ehrlich tumour and studied in a comparative manner in relation to that found in mice plasma. Enzymes from both sources were characterized with respect to optimum pH, substrate concentration and quinidine inhibition. After gel filtration by Sephadex G-200 and Sepharose 6B, two enzyme forms were observed in ascitic fluid as well as in mice plasma: a large form (L) and a small form (S) presenting molecular weights of 191 000, and 224 000 daltons for L forms and 71 000 and 69 000 daltons for S forms respectively. Concanavalin A interacts with both molecular forms, suggesting a glycoprotein nature for these enzymes.     

Morphological changes in frog pronephric cell surfaces after transformation by herpes virus.

A scanning electron-microscope study has established that there are major modifications in the surface membranes of malignant cells transformed from the pronephric kidney of the frog. Tumours were induced in vivo after exposure of embryos Lucké Herpes virus. A pronephric cell line from normal embryos and 3 tumour cell lines derived from experimentally induced adenocarcinomas of the pronephros were observed. The normal embryonic kidney cells have their surfaces covered with long fingerlike microvilli throughout the cell cycle. In contrast, the surface of the pronephric tumour cells are mainly covered by multiple, ruffled or parallel straight microridges and short, stubby microvilli. These comparisons were made on cells subject to similar culture conditions. The topographic changes in the tumour cells are considered significant since the characteristic microridges and stubby villi were consistently found in all 3 pronephric tumour lines.     

Prostaglandin Production by Experimental Tumours and Effects of Anti-inflammatory Compounds

THE presence of small quantities of prostaglandin-like material has been demonstrated in medullary carcinoma of the thyroid¹ and in Kaposi's sarcoma²; the diarrhoea often associated with these tumours has been attributed to prostaglandins. Diarrhoea is also often present in mice bearing the BP8/P₁ tumour. Preliminary investigations of the ascitic fluid from mice inoculated with this tumour showed the presence of pharmacologically active substances and prostaglandin E₂-like activity was found in small amounts³. Subsequent examination of the BP8/P₁ cells has revealed concentrations of prostaglandin E₂ in excess of 100 times that of the fluid. Significant amounts have also been found in another tumour, sarcoma 180 (S180).     

Specific chalone inhibition of the regeneration of the JB-1 ascites tumour studied by flow microfluorometry.

The variation in the DNA distribution in the JB-1 and the Lla2 ascites tumour was investigated by means of flow microfluorometry (FMF) in the plateau stage and during the initiation of the regenerative growth induced by percutaneous aspiration. The study showed that a considerable influx of cells with G1DNA content into the S phase occurred in both tumours about 10 hr after aspiration. In the JB-1 tumour, these initial regenerative changes could be reversibly blocked by injections of cell-free plateau JB-1 ascitic fluid or an ultrafiltrate of this ascites. In contrast to these observations no delay in the regenerative changes was observed in the L1a2 tumour after treatment with JB-1 ascites or the ultrafiltrate. The study supports the assumption of a specific growth regulation of the JB-1 ascites tumour and emphasizes the suitability of FMF analyses in cell-kinetic studies in which short-term fluctuations take place in the distribution of cells with different DNA content.     

The modelling of the growth of Ehrlich carcinoma and teratoma T-36 with ascitic fluid and tumor-cell dialysate]

The study of the effect of ascitic fluid and dialysate of Ehrlich ascites tumor cells (M.m. less than 15 kDa) on the growth of Ehrlich carcinoma and teratoma T-36 has shown that both the ascitic fluid and dialysate can protect tumor cells in vivo. The number of animals with tumors increased from 0% in control animals to 60 and 20%, respectively, in experimental ones after transplantation i.m. of 20 x 10(3) Ehrlich tumor cells into mice. Compared to control, ascitic fluid and dialysate of Ehrlich ascites tumor cells increased the rate of tumor growth to 195 and 153%, respectively. It is suggested that this test-system simulates the effect of tumor humoral factors in vivo.