PubMed Health⌕ Search

Biomedical subjects

G Starace

Publications and source records attributed to G Starace.

At least 37 records · Page 2Linked to original sources
PubMed1994

Interferon gamma up-regulates alpha 2 macroglobulin expression in human astrocytoma cells.

An established human astrocytoma cell line (T67) was shown to constitutively produce the proteinase inhibitor alpha 2 macroglobulin (alpha 2M). Interferon gamma (IFN gamma), a potent immunoregulatory lymphokine, was able to increase the synthesis of alpha 2M by these cells, as measured by ELISA on cell supernatants. The alpha 2M induction was also observed in other human glioma cell lines (T70 and ADF) and in human fetal astrocyte cultures following IFN gamma treatment. In T67 cells this effect was dose-dependent and the maximum (2.7-fold increase) was obtained with 2000 U/ml of IFN gamma. A corresponding enhanced alpha 2M mRNA accumulation was demonstrated by PCR and Northern blot techniques. Our results suggest an important role of alpha 2M during inflammatory and immune processes in the CNS. An increased release of alpha 2M following IFN gamma stimulation may allow the removal of the bulk of proteases released at the site of inflammation, strengthening at the same time the antigen presentation processes.

Astrocytoma↗
PubMed1993

A stochastic model for cell debris in flow cytometry.

An important problem in analyzing flow cytometry DNA measurements, especially for tumor samples, is the presence of background noise, mainly resulting from cellular debris. Several models have been proposed for subtracting this contribution from DNA content histograms. In the present paper we propose a model for background debris distribution based on a specific mechanism for the DNA fragmentation process of the cell nucleus. In particular, we suggest that the fragmentation is more likely to occur in the sites which are closer to the end points of the DNA chain. This accounts for the observed higher frequency of small-sized fragments in experimental DNA distribution. The proposed model depends on two parameters describing the intensity and the shape of the background. It has been successfully validated against four series of experimental data of DNA distribution with increasing level of background.

Cell Fractionation↗
PubMed1993

Receptor-mediated endocytosis of alpha 2 macroglobulin by a glioma cell line.

Previous experiments have shown that human neoplastic and embryonic glial cell lines synthesize and secrete in culture, alpha 2 macroglobulin (alpha 2M), a broad spectrum proteinase inhibitor present in serum and extracellular fluids. The present study was aimed to investigate the presence of alpha 2M receptors on glial cell membrane, since several non-neural cell types producing alpha 2M also express alpha 2M receptors. By flow cytometric analysis, immunofluorescence and immunoelectronmicroscopy techniques we demonstrate an alpha 2M receptor-related immunoreactivity on the plasma membrane of a human glioma cell line. Ultrastructural experiments reveal a close colocalization of immunoreactivities for alpha 2M and its receptor in clathrin-coated pits and vesicles, structures typically involved in receptor-mediated endocytic pathways.

Endocytosis↗
PubMed1993

Ras oncogene transformation of human B lymphoblasts is associated with lymphocyte activation and with a block of differentiation.

The p21ras small GTP binding proteins participate in signal transduction from cell surface receptors and affect neoplastic transformation and development in many different cell types. In the present study, we examined the relationship between ras transformation and differentiation of human B lymphocytes. We show that the constitutive expression of the T24 Ha-ras oncogene in EBV-immortalized B lymphoblasts was associated with the induction of the interleukin 2 receptor alpha subunit, with an impaired immunoglobulin gene expression, altered adhesion properties and increased survival in serum-free medium. Since induction of the IL-2 receptor alpha subunit is a hallmark of lymphocyte activation, we suggest that p21ras naturally triggers B cell activation. The ras-transformed lymphocytes displayed a fully functional IL-2r, as assessed by c-fos induction following treatment with IL-2; nevertheless, they were not growth stimulated by this lymphokine. The decreased expression of immunoglobulin genes indicates that the ras oncogene blocks terminal differentiation to plasma cells, possibly by inhibiting the activity of lymphocyte-specific transcription factors. Somewhat unexpectedly, the constitutive p21ras activity did not cause an increased DNA binding of transcription factors PEA1 (AP1), PEA3, Oct-2 or NF-kB.

B-Lymphocytes↗
PubMed1992

Immunosuppressive molecules produced by Sertoli cells cultured in vitro: biological effects on lymphocytes.

In the present study we have analyzed the proteins secreted in vitro by murine Sertoli cells to identify immunosuppressive factors. Our data show that Sertoli cells secrete molecules capable to inhibit proliferation of lymphocytes activated in vitro. Cytophluorimetric analysis indicates that treated cells are arrested in the G1 phase of cell cycle. The inhibitory activity is specific for both B or T lymphocytes but not for other non-lymphoid cells and is associated to proteins, heat and freeze stable, with Mr of more than 30 kDa. Lymphocytes treated with Sertoli immunosuppressive proteins drastically reduce the secretion of interleukin-2.

Animals↗
PubMed1992

Fc gamma receptors are expressed on human neuroblastoma cell lines: lack of correlation with N-myc oncogene activity.

FcRs (Fc Receptors) have been detected on the cell surface of two human neuroblastoma cell lines; IMR 32 and SK-N-SH, by immunocytochemistry and flow cytometric analysis, using a previously characterized polyclonal antiserum raised against the Fc gamma R isolated from a human CLL line (Gorini, Medgyesi, Garavini, Dorrington and Down, 1987; Rozsnay, Sarmay, Szabo, Medgyesi, Gorini and Gergely, 1990). FcR is expressed on all the cells of both lines at least at the same level as on the HL60 promyelocyte cell line used as positive control. Two electrophoretic components displaying apparent molecular masses of 70 and 43 kDa respectively have been identified by SDS-PAGE followed by Western blotting analysis of crude cell membranes. In addition, "in situ" hybridization experiments seem to exclude a correlation between FcR expression and N-myc oncogene activity. The presence of FcR in neuroblastoma could be related to a possible functional role even on these cells which do not belong to the immune system; moreover, they could also be exploited for a diagnostic characterization of this tumor.

Blotting, Western↗
PubMed1990

Study of propidium iodide binding to DNA in intact cells by flow cytometry.

We studied the in situ binding of propidium iodide to DNA in fixed human lymphocytes, using flow cytometry. Experimental data of fluorescence emission vs dye concentration and vs cell concentration were obtained. Data were interpreted by means of two different mathematical models specific for the staining reaction, and the binding parameters were obtained by "best-fitting" of the data. A model based on two classes of binding sites with different affinity constants gave the most satisfactory fitting. The accessibility of the in situ chromatin turned out to be reduced with respect to the non in situ accessibility for ethidium bromide as reported in the literature. The present study shows the usefulness of the flow-cytometric technique for probing DNA structure in intact cells.

Binding Sites↗
PubMed1987

Kinetic and survival response of the M14 cell line to lonidamine associated with adriamycin or hyperthermia.

Lonidamine (LND), an indazole-carboxylic acid derivative, was delivered alone and together with adriamycin (ADM) or hyperthermia to the human melanoma cell line M14, and cell survival was assessed. Cell cycle-specific effects were investigated by analyzing sequences of DNA content histograms by means of a suitable mathematical procedure. LND delivered for 1 h at a dose of 50 micrograms/ml did not affect proliferation and survival of the cells. Exposure of the cells for 1 h to ADM (1.0 microgram/ml) followed by LND for 1 h (50 micrograms/ml) produced the highest effect on the survival. Kinetic parameters were affected by the combined treatment slightly more than by ADM exposure alone. Simultaneous delivery of LND (50 micrograms/ml) with hyperthermia (42 degrees C, 1 h) reduced the survival and enhanced the block of cells in the G2M phase, as compared with the heat treatment alone. The effect of the treatments on cell survival appeared to be related to the perturbation of the G2M phase of the cycle.

Antineoplastic Agents↗
PubMed1986

Cyclosporin A antileukemic activity: mode of action on target cells.

Cyclosporin A (CyA) is a potent immunosuppressive agent which has been reported to inhibit proliferation of human T leukemic lymphoblasts. We used the permanent cell line 8402 to perform a detailed investigation of the CyA mode of action on such cells. CyA treatment resulted in a dose-dependent inhibition of the 8402 cells growth rate which was paralleled by a decrease of tritiated thymidine uptake. These findings were the consequence of redistribution of treated cells in the cell cycle phase. By means of intracellular DNA content measurements made by flow cytometry, it was shown that the primary effect of CyA was accumulation of treated cells at the G1 phase of the cell cycle. At the doses used, CyA activity was not spontaneously reversible, resulting in death of treated cells which was apparent at about 72 h after exposure to the drug.

Cell Cycle↗
PubMed1984

Analysis of DNA synthesis rate of cultured cells from flow cytometric data.

The rate of DNA synthesis along S phase is estimated from flow cytometric histograms on the basis of a mathematical model of a cell population. In the absence of loss, the model expresses the population kinetics in terms of DNA synthesis rate, S-phase influx, and population size. A single histogram is sufficient to determine the DNA synthesis rate when the population is in balanced exponential growth. Two suitably chosen histograms are necessary if the S-phase influx is exponential in a time interval longer than the S-phase duration. The analysis procedure was tested on published autoradiographic data and applied to three cultured cell lines (CM-S, 3LL, and M14 cells) that show various patterns of DNA distribution. In each case the cell-cycle fractions, the DNA synthesis rate, and the S-phase duration were obtained.

Animals↗
PubMed1983

Glucocorticoid receptors and cortico-sensitivity in a human clonal monocytic cell line, CM-SM.

CM-SM is a clonal line of human precursor mononuclear phagocytes inducible to macrophage differentiation in response to the tumor promoter phorbol ester 12-O-tetradecanoyl-phorbol-13-acetate (TPA). Untreated CM-SM cells contain single class, high-affinity (KD = 4.0 X 10(-9) M) glucocorticoid-specific receptor sites (approximately 60,000 per cell), as measured by a whole cell assay, at 37 degrees C, using [3H]triamcinolone acetonide (TA). Exposure of CM-SM to dexamethasone (DEX) produced a progressive, dose- and time-related series of changes in CM-SM cell growth, saturation density, morphology, and functional properties, with half-maximal effects at about 10(-9) M for DEX. TA-receptor sites rapidly decreased (about 70%) after DEX treatment, without any apparent change in steroid specificity and affinity. After 5 days in culture with a saturating concentration (3.6 X 10(-8) M) of hormone, the cells reached a saturation density of about 9.0 X 10(6) viable cells/ml (about 4.0 X 10(6) viable cells/ml in the controls), while the modal volume of the resulting cell population was approximately 60%, as compared to the volume of untreated cells. DEX-treated cells appeared less differentiated than controls, as assessed by combined morphologic, antigenic, and cytoenzymatic analyses. DEX almost completely inhibited TPA activation of the following macrophage functions: adherency to the culture plate, expression of lysosomal enzymes, Fc and C3 receptors, and stimulation of phagocytosis. After removal of DEX, the cells, within a few passages, returned to a state apparently identical to the untreated controls and could be induced to macrophage differentiation in response to TPA.

Cell Differentiation↗
PubMed1982

Flow cytometric analysis of cellular DNA in human acute nonlymphatic leukemias and dysmyelopoietic syndromes.

The proliferation patterns of bone marrow cells from 16 cases of acute nonlymphatic leukemia (ANLL) and 13 of dysmyelopoietic syndromes were analyzed with DNA-flow cytometry. In order to reduce the contamination by nonproliferating peripheral blood cells, all aspirates were submitted to a density centrifugation step. DNA content measurements were performed with a PHYWEICP 22 flow system apparatus. The distribution of cells in te cell cycle phases was calculated by means of an automated fitting procedure. No significant difference between the cell cycle distribution in ANLL and dysmyelopoietic syndromes was detectable. No relationship was observed between the size of the S phase compartment and the percentage of blasts in the original samples. Aspirates from ANLL were also cultured in vitro in liquid phase on dialysis membranes. A clear relationship was observed between the size of the phase compartment and in vitro cell growth since, with the exception of one case, all samples with less than 11% cells in S phase failed to proliferate in vitro. The proliferation profile of ANLL cells after 10 days in culture was similar to that of the original samples.

Anemia, Aplastic↗
PubMed1982

Cytometric evidence of cytogenetic and proliferative heterogeneity of human solid tumors.

Flow cytometry is a widely recognized method of rapidly assessing the ploidy and proliferation status of experimental and solid tumors. In the present work, a variety of human cancers from various sites (lung, head and neck, etc.), of traditional interest in our laboratory, have been analyzed. In agreement with a number of recent reports, a general heterogeneity of human solid tumors can be evidenced. In particular: (a) solid tumors are characterized by a variable degree of aneuploidy; (b) the internal structure of solid tumors is highly heterogeneous especially with respect to the fraction of aneuploid malignant cells and their distribution through the cycle phases; and (c) some solid tumors are also characterized by the presence, to a variable extent, in the tumor of mass of multiple cell clones. Static fluorimetry of Feulgen-stained (mitotic) single cells offers a way to confirm this kind of observations.

Aneuploidy↗
PubMed1982

DNA content distribution of in vivo and in vitro lines of Lewis lung carcinoma.

The kinetic features of Lewis lung carcinoma (3LL) and of two in vitro and two in vivo derivative lines were studied by flow cytometry. The in vitro lines C108 and BC215 show the same tetraploid DNA content and a very similar cell cycle structure, characterized by a prevalent S fraction. Also, the in vivo lines, the original 3LL and M1087, show a tetraploid DNA content, while the BM21548 is characterized by a hyperdiploid DNA mode and a broader distribution of DNA values. No difference in the modal DNA value is found between each primary tumor and the corresponding lung metastases for all the in vivo lines. In addition, an increase in the G1 component corresponding to a decrease in the S fraction is observed during the tumor growth. These kinetic features were related to some malignant properties of 3LL lines, such as growth pattern and metastatic potential. Our findings indicate that a direct correlation is not always possible to establish.

Animals↗
PubMed1982

Early effects of chemical carcinogens as compared to induced cell proliferation. I. Flow microfluorimetry.

Acridine orange-stained rat liver cells have been analyzed by flow microfluorimetry either after i.p. injection of 0.6 and 5.4 mg/kg of dimethylnitrosamine or 18 hr after partial hepatectomy. Treatment with dimethylnitrosamine induced a marked increase in the fraction of cells with higher green fluorescence intensity which can be related to a greater number of chromatin primary binding sites, and a decrease in the fraction of cells with higher red fluorescence intensity corresponding to a lower amount of cellular RNA. Contrary to the carcinogen-induced early changes towards a more relaxed uncoiled structure, as well as an apparent uncoupling between chromatin-DNA structural alterations and total RNA synthesis, could be first events in the carcinogenetic process.

Acridine Orange↗