SV40-3T3 cells were exposed in monolayer cultures to 5 X 10(-7) M methotrexate (MTX), that inhibited thymidylate synthetase, arrested cell growth without cell killing in 24 h and did not induce single- (ss) or double-strand (ds) breaks in DNA. Following 24, up to 72 h, the poly(ADP-ribose) polymerase content of ...

The MFO activity in intact cells was monitored through microspectrofluorimetry study of BaP metabolization. This approach allowed observation of individual cells in a cell population to represent the studied samples through histograms as a function either of rate constants or of "residual BaP". The inhibitory effect of ellipticine was demonstrated ...

In this study, we have demonstrated that a murine T cell lymphoma, EL 4, and a murine fibroblast cell line, Swiss 3T3, possess a single class of high affinity interleukin 1 (IL 1) receptors that exist in a dynamic state of equilibrium that is influenced by IL 1. In the ...

Fluorescence ratio imaging microscopy (Tanasugarn, L., P. McNeil, G. Reynolds, and D. L. Taylor, 1984, J. Cell Biol., 98:717-724) has been used to measure the spatial variations in cytoplasmic pH of individual quiescent and nonquiescent Swiss 3T3 cells. Fundamental issues of ratio imaging that permit precise and accurate temporal and ...

Quiescent, chemically transformed (benzo-alpha-pyrene) BALB/c 3T3 cells (BP A31) enter the cell division cycle when exposed to complete medium containing 10% fetal calf serum (FCS); the number of cells recruited is a function of the duration of serum exposure. The recruitment of cells by short (less than 4 h) serum ...

Fibroblasts derived from patients with Werner's syndrome (WS) were incubated with radioactive sulfate to study the incorporation of 35S into glycosaminoglycans (GAGs). The accumulation of cell-associated 35S radioactivity in the GAGs of WS fibroblasts was consistently higher than parallel accumulation in normal human fibroblasts, but was substantially less than in ...

A quantitative analysis based on centrifugal force requirements for enucleation was developed to examine the response of a number of untransformed and transformed cell lines to cytochalasin mediated enucleation. Examination of the extent of cell enucleation as a function of centrifugal force resulted in a series of response curves demonstrating ...

Mouse 3T3 cells were transformed with an antisense c-fos gene fused to a mouse mammary tumor virus promoter. In transformants that integrated a large number of antisense c-fos sequences, the usual large increase in c-fos mRNA and protein following stimulation of quiescent cells by platelet-derived growth factor was blocked in ...

The B subunit of cholera toxin, which binds specifically to several molecules of ganglioside galactosyl-(beta 1----3)-N-acetylgalactosyminyl(beta 1----4)-[N- acetylneuraminyl(alpha 2----3)]-galactosyl(beta 1----4)glucosyl(beta 1----1) ceramide (GM1) on the cell surface, stimulated DNA synthesis and cell division in quiescent, nontransformed mouse 3T3 cells in a dose-dependent manner. In addition, the B subunit potentiated the ...

Ribonucleotide reductase from mammalian cells is composed of two nonidentical subunits M1 and M2 which are both required to form the catalytic site. The level of ribonucleotide reductase activity is cell cycle controlled and several reports suggest that this control is achieved mainly by the regulation of M2 subunit synthesis. ...

The mouse neuroblastoma x rat glioma hybrid cell line, NG108-15, does not synthesize serotonin from tryptophan, although the cells take up tryptophan and serotonin in a saturable manner from serum-supplemented incubation medium. Since serum commonly used to supplement the growth medium contains serotonin, it is concluded that appreciable levels of ...

We have isolated a syngeneic monoclonal antibody (HepSS-1) reactive to a murine methylcholanthrene-induced fibrosarcoma, Meth-A. HepSS-1 also bound to a wide variety of established and fresh normal cells derived from not only mice but also other species such as human, monkey, rat, hamster, and chicken. Immunoprecipitation of surface iodinated Meth-A ...

A helical mycoplasma, Spiroplasma mirum strain SMCA, produced malignant transformation in mouse NIH 3T3 cells and monkey kidney CV-1 cells. The transformed cells exhibited morphological changes consistent with the transformed phenotype, grew in soft agar and produced tumors in athymic and BALB/c mice. Transmission electron microscopy revealed structures morphologically similar ...

Analysis of the responsiveness of isolated adult urothelium to a series of different stromal cell-extracellular matrix combinations demonstrated the capacity of stromal cells to induce and maintain normal patterns of urothelial growth, differentiation, and maturation in vitro. By incorporating embryonic mesenchymal derived (Swiss 3T3) cells into type I collagen matrices, ...

Statin, a protein of 57,000 D, is present in the nuclei of quiescent or senescent fibroblasts (Wang, E, J cell biol 100 (1985) 545), but is absent in their young replicating counterparts. Immunohistochemical survey of a variety of tissues demonstrates that the presence of statin is a marker for cells ...

The effect of phenobarbital upon the differentiation of two preadipocyte cell lines, 3T3 F442A and 3T3 L-1, was examined by measuring the synthesis and secretion of lipoprotein lipase. Extracellular enzyme was measured by treating intact cells with heparin, and the intracellular enzyme was subsequently assayed in cell homogenates. When confluent ...

Neutron Activation Analysis (NAA) was used to investigate trace element patterns in serum, packed cells, and dialysate of CAPD patients. The concentrations of the elements Cs, Cu, Fe, and Mn in serum and packed cells appeared to be maintained within the normal range, while the levels of the non-essential element ...

The effect of inhibition of 3-Hydroxy-3-methylglutaryl Coenzyme A reductase (HMG CoA reductase) on cell cycle progression in proliferating 3T3 cells was studied. It was found that short transient exposures to the HMG CoA reductase inhibitor 25-hydroxycholesterol temporarily blocked the cell cycle traverse in the postmitotic half of G1 (G1pm), whereas ...

Tubulin expression was analysed in normal and simian virus-40 (SV40) transformed 3T3 cells by two-dimensional polyacrylamide gel electrophoresis and immunoblotting studies using monoclonal antibodies raised to alpha- and beta-tubulin subunits. The ratio of alpha- to beta-tubulin recognised was calculated for both cell lines and found to shift from 2.50 in ...

A bovine sialoglycopeptide, purified to homogeneity and capable of inhibiting cellular protein synthesis and proliferation, was shown to agglutinate a wide variety of nontransformed and transformed cells. The cell agglutination activity was shown to be independent of the biological inhibitory action and most likely related to a protease activity that ...

The ability of the calcium ionophore A23187 and the sodium ionophore Monensin to antagonize the inhibition of 3T3 cell protein synthesis by a bovine cell surface sialoglycopeptide was measured. A23187, when added before and shortly after the sialoglycopeptide, significantly reduced the biological activity of the inhibitory glycopeptide. In contrast, Monensin ...

Proliferating nonconfluent 3T3 cells become committed to proceed through the cell cycle or to enter G0 during the first post-mitotic part of G1 (G1pm). The decision to proceed through G1pm is dependent on the presence of serum growth factors in the culture medium. Cells that have passed this particular growth-factor-dependent ...

The proteoglycans secreted by a malignant human breast cell line (MDA-MB-231) were compared with the corresponding proteoglycans from a normal human breast cell line (HBL-100). The physicochemical characteristics of these proteoglycans were established by hexosamine analysis, chemical and enzymatic degradations, and dissociative cesium chloride density gradient centrifugation, and by gel ...

To understand the earliest phases of epidermal cell spreading we have sought a defined in vitro system. We studied the divalent cation dependence of guinea pig epidermal cell spreading in media containing varying concentrations of cations. No spreading occurred in calcium-magnesium-free Dulbecco's modified Eagle's medium (CMF-DME) in the presence of ...

The observed homology between G-proteins which regulate adenylate cyclase and ras proteins and the suggested role of ras in the regulation of adenylate cyclase in yeast prompted us to examine the regulation of adenylate cyclase in three cell lines: (i) NIH 3T3 cells, (ii) NIH 3T3 cells transformed by high ...

When single cells were allowed to attach to circular islands of adhesive substratum, their proliferation was strongly dependent on island area over the range 500 micron2 to 5000 micron2. The number of freshly explanted whole mouse embryo fibroblasts that performed DNA synthesis corresponded closely with a simple geometrical measure of ...

A sialoglycopeptide inhibitor, isolated from bovine cerebral cortex cells, that reversibly inhibits protein and DNA synthesis, was coupled to either Sepharose or polyacrylamide beads. Whereas over 1 ng of the inhibitor was released from Sepharose beads after 30 min at 37 degrees C, less than 0.2 ng of the sialoglycopeptide ...

The relationship between exposure to asbestos fibers and the development of mesothelioma or bronchial carcinoma prompted many countries to ban its use from commercial products. The biological mechanism by which asbestos induces or promotes mesothelioma or carcinoma is still unknown. In order to study the influence of fibers on the ...

Using metallic gold in various assays for the motility of cultured tissue cells, the paper compares the movements of surface projections and the locomotion of polyoma (Py3T3) and SV40 (SV3T3) virus-transformed 3T3 cells with the behavior of the parental 3T3 cells. The movement of surface projections was assayed by the ...

In the mixed culture system nontransformed 3T3 fibroblasts cells inhibited the plating efficiency of neuroblastoma (N2a) cells, whereas heat or methanol killed fibroblasts did not. In contrast, chloroform/methanol extracted 3T3 cells inhibited the plating efficiency of neuroblastoma cells to the same extent as had been the case with living 3T3 ...

Addition of aluminum to the culture medium of Nakano mouse lens epithelial (NMLE) cells and Swiss 3T3K cells induced both 3H-thymidine incorporation and mitosis. This is in contrast to other metal ions such as vanadium, which, at concentrations high enough to increase 3H-thymidine incorporation, actually inhibits mitosis (Jones and Reid, ...

Neonatal rat heart cells cultivated in either of two different media which varied only in their serum supplements were transferred to chemically defined medium (Ham's F10) for 24 h before measuring a variety of parameters. The 24-h period of exposure to chemically defined medium was not sufficient to reverse the ...

Foetal bovine serum (FBS) and the preparation Ultraser (LKB, Sweden) contain more growth factors (GF) necessary to promote proliferation of cells in soft agar than the sera from newborn and adult animals. It was found that non-dialysable GFs are produced during growth of cells in semisolid medium. The dependence of ...

We have isolated and characterized glycopeptides, derived from mouse and bovine cerebral cortex cells, that inhibit protein synthesis and cell growth of normal but not transformed cells. The inhibitor binds to target cell surfaces, and gangliosides have previously been shown to influence cell sensitivity to the glycopeptides. Preincubation with 3.0 ...

Inositol lipid turnover was studied in quiescent Swiss mouse 3T3 cells stimulated by platelet-derived growth factor (PDGF). Stimulation of the cells by PDGF for 10 min at 37 degrees C induced the following changes in lipids: in cells prelabelled with [32P]Pi, a 28% decrease in [32P]phosphatidylinositol 4,5-bisphosphate, a 41% decrease ...

The relation between cellular rRNA content, as a measure of cell size, and the regulation of the cell cycle has been investigated for swiss 3T3 and the spontaneously transformed swiss 3T6 cell line. It is shown that the characteristic of percent of quiescent cells stimulated into the cell cycle versus ...

Resident peritoneal mouse macrophages (non-dividing differentiated cells) were fused with mouse embryo fibroblasts (cells with a limited lifespan), NIH 3T3 and C3H 10T 1/2 cells ('immortal' cell lines) and SV 3T3 cells (a malignant cell line). DNA synthesis was investigated in the resultant heterokaryons. No inhibitory effect upon the transition ...

Reproducible conditions were defined for using rates of leucine incorporation as a valid measure of rates of de novo protein synthesis in mouse 3T3 cells. Upon stimulation of quiescent cultures, rates of de novo synthesis of proteins increased and pool levels of amino acids decreased in proportion to the concentration ...

The report of Carrel and Ebeling (J. Exp. Med., 34 (1921) 599-623) generally gives the impression that both serum and blood plasma from old animals inhibit cell proliferation. For confirmation of this, we examined the effects of serum from rabbits of various ages on rabbit fetal skin fibroblasts (RSF cell) ...

A simple endpoint dilution method for evaluating foetal calf serum quality is described. The test uses a series of doubling dilutions of cells on microtitre trays with the test sera added to replicate dilution series. After five to six days of incubation the cells are stained with crystal violet and ...

Rabbit antibodies raised against microtubule-associated protein 1 (MAP-1) from hog brain were found to crossreact with extracellular matrix components of mouse BALB/c 3T3 cell cultures. As shown by immunofluorescence microscopy of confluent cell cultures, the extracellular MAP-related antigen was located on dense fibrillar network arrays underlying and surrounding the cells. ...

We have confirmed the reports of Harper and Grove (Science, 204 (1979) 526-527), and Azzarone and Macierira-Coehlo (J. Cell Sci., 57 (1982) 177-187) that fibroblasts derived from the papillary dermis have greater in vitro growth potential and longer replicative lifespans than genomically identical fibroblasts derived from the reticular dermis. In ...

We report here a unique system for tracking normal human urothelial cell migration in serum-free culture medium (HMRI-1). The key observation was that urothelial cells deposited red blood cell surface antigen on the culture dish in a remarkable pattern. Scrutiny of this pattern showed that each migrating cell left behind ...

The regulation of ppp(A2'p)nA-(2-5A)-dependent RNase (RNase L or RNase F) was investigated in NIH 3T3, clone 1 cells using 2-5A-binding and nuclease activity assays. Minimal levels of 2-5A-dependent RNase were detected in actively dividing clone 1 cells; these levels were independently induced by growth arrest or interferon treatment. Accordingly, levels ...

Kinetic analysis of cellular response to serum deprivation or inhibition of protein synthesis was performed on Swiss 3T3 cells. Time-lapse cinematographic analysis of individual cells transiently exposed to serum-free medium (with or without the addition of purified growth factors) or cycloheximide enabled a detailed mapping of the magnitude and variability ...

The role of phenomena analogous to fibroblast contact inhibition in lymphocyte growth regulation is controversial, although it is clear that direct cell-cell contact is vital to immunoregulation and accessory cell function. An extract of mouse liver plasma membrane proteins, referred to as suppressive liver extract (SLE), that suppresses the growth ...

The structural and biological properties of a polyomavirus mutant selected in Friend erythroleukemic cells were investigated. The growth efficiency of this mutant (PyFL78) was compared with that of the parental PyA2 strain by a growth competition assay in Friend erythroleukemic and 3T3 (or 3T6) cell lines. The results reveal that ...

10-[Diethylaminopropylamino]-6-methyl-5H-pyrido[3',4':4,5] pyrrolo[2,3-g]isoquinoline (BD-40) (NSC-327471D) is an aza-ellipticine derivative with a promising antitumor activity (M. Marty, C. Jasmin, P. Pouillard, C. Gisselbrecht, G. Gouvenia, and H. Magdalainat, 17th Annual Meeting of the American Society of Clinical Oncology, C-108, 1981) and less toxicity than ellipticine. We have compared the effects of ellipticine, ...

The effect of submicromolar concentrations of the K+ ionophore valinomycin on proliferation, viability, distribution of cell population over phases of the cell cycle, and cellular adenosine triphosphate content of different permanent rodent cell lines in vitro was investigated. Valinomycin inhibits proliferation of all cell lines tested with a saturating effect ...

We examine changes in protein synthesis that accompany suspension (i.e. shape alteration) of anchorage-dependent and anchorage-independent cells using a newly developed cell fractionation procedure based on detergent extraction. Using this procedure, a cell can be divided into four distinct and independent fractions: soluble, cytoskeleton, chromatin and nuclear matrix-intermediate filament. This ...