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  • 1
    Electronic Resource
    Electronic Resource
    Final checkpoint in the drug-promoted and poliovirus-promoted apoptosis is under post-translational control by growth factors (1996)
    New York, N.Y. : Wiley-Blackwell
    Journal of Cellular Biochemistry 63 (1996), S. 422-431 
    Details
    ISSN: 0730-2312
    Keywords: actinomycin D ; cycloheximide ; DNA degradation ; chromatin fragmentation ; serum factors ; epidermal growth factor ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology , Medicine
    Notes: The treatment of HeLa subline (HeLa-B) cells with cycloheximide or Actinomycin D resulted in a rapid (∼ 1.5 h and ∼ 2.5 h, respectively) development of morphological and biochemical signs of apoptosis. The addition of fetal bovine serum to the cycloheximide-treated or Actinomycin D-treated cells suppressed the apoptotic reaction, as evidenced by the postponement of the DNA fragmentation for at least 9 and 5 h, respectively. A similar suppressive effect was observed upon the serum addition to cells undergoing abortive infection with poliovirus, which died of apoptosis in the absence of the serum. The serum appeared to exert its anti-apoptotic effect without any appreciable lag and even immediately blocked further progress of ongoing DNA fragmentation. The epidermal growth factor also suppressed, although less efficiently and more transiently, the apoptotic reaction promoted by the metabolic inhibitors. It is concluded that growth factors may affect, without modulating either transcription or translation, the balance of pro-apoptotic and anti-apoptotic activities at a final checkpoint, just preceding the irreversible effector step of apoptosis. © 1996 Wiley-Liss, Inc.
    Additional Material: 5 Ill.
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  • 2
    Electronic Resource
    Electronic Resource
    Proton production and consumption pathways in yeast metabolism. A chemostat culture analysis (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Yeast 11 (1995), S. 1353-1365 
    Details
    ISSN: 0749-503X
    Keywords: yeast ; nitrogen pathway ; chemostat culture ; proton production ; pH ; metabolic model ; control ; Life and Medical Sciences ; Genetics
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: In this investigation, a method for the accurate quantitative determination of net proton production or consumption in biological cultures has been devised. Cells are cultured under constant pH conditions. The specific rate of proton production or consumption by the culture (qH+, mmol h-1 per g biomass) is proportional to the mmol of base or acid required to maintain constant pH per unit time, and this equivalence is independent of the buffering capacity of the culture medium.The above method has been applied to chemostat cultures of Candida utilis growing on glucose or glycerol as carbon source, and different nitrogen sources. The results indicate that the nitrogen assimilation pathway alone determines the value of qH+, and a fixed stoichiometric relationship between nitrogen uptake rate qN (meq h-1 per g biomass) and qH+ has been found for each nitrogen source employed. Thus, qH+/qN values of +1, 0 and - 1 were found for ammonium ions, urea and nitrate respectively. Under oxidative metabolism, the contribution of carbon catabolism to the value of qH+ was undetectable.Since qN may be related to growth and production of type 1 compounds in fermentation processes, the parameter qH+ was incorporated into a model of growth and energy metabolism in chemostat culture (Castrillo and Ugalde, Yeast 10, 185-197, 1994), resulting in adequate simulations of experimentally observed culture performance. Thus, it is suggested that qH+ may be employed as a simple and effective control parameter for biotechnological processes involving biomass-related products.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/yea.320111404
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  • 3
    Electronic Resource
    Electronic Resource
    Role of the cytoskeleton in the reaction of fibroblasts to multiple grooved substrata (1995)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 31 (1995), S. 147-158 
    Details
    ISSN: 0886-1544
    Keywords: actin ; contact guidance ; microfilaments ; microtubules ; orientation ; cytochalasin ; colcemid ; taxol ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The role of the cytoskeleton and cell attachments in the alignment of baby hamster kidney fibroblasts to ridge and groove substratum topography was investigated using confocal scanning microscopy. This was carried out with normal cells and cells treated with the cytoskeleton modifiers cytochalasin D, colcemid, and taxol. Actin was localised with fluorescent phalloidin. Tubulin, Vinculin, and intracellular adhesion molecule-1 were visualised by indirect immunofluoresence. The spreading, elongation, and orientation of the cells after 24 h of culture in these conditions were measured on grooves of 5, 10, and 25 μm width and 0.5, 1, 2, and 5 μm depth. We have also observed events over the first 30 min of cell attachment. Five minutes after cell attachment, F-actin condensations were seen close to the intersection of groove wall and ridge top, that is, at a topographic discontinuity. The condensations were often at right angles to the groove edge and showed a periodicity of 0.6 μm. Vinculin arrangement at the early stages of cell spreading was similar to that of actin. Organisation of the microtubule system followed later, becoming obvious at about 30 min after cell plating. The Curtis and Clark theory (that cell react to topography primarily at lines of discontinuity in the substratum by actin nucleation) is supported by these results. The use of cytoskeletal poisons did not entirely abolish cell reaction to grooves. Colocemid increased cell spreading and reduced cell orientation and elongation. Cytochalasin D reduced cell spreading, orientation, and elongation. Taxol reduced cell elongation but did not affect cell spreading and orientation. We conclude that the aggregation of actin along groove/ridge boundaries is a primary driving event in determining fibroblast orientation on microgrooved substrata.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970310207
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  • 4
    Electronic Resource
    Electronic Resource
    Focal adhesion formation is associated with secretion of allergic mediators (1995)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 31 (1995), S. 215-224 
    Details
    ISSN: 0886-1544
    Keywords: RBL-2H3 cells ; vinculin ; mast cells ; talin ; cytoskeleton ; permeabilized ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Adherence of cells to the extracellular matrix via focal adhesions is known to modulate many cellular functions. However, the role of focal adhesions in the regulation of secretion is unclear. To examine this we have used the RBL-2H3 rat mast cell line, in which we and others have observed cytoskeletal rearrangements and increased cell spreading during secretion. All activators of secretion examined, whether acting specifically through or bypassing the IgE-receptor, induced the assembly of focal adhesions, as defined by the localization of vinculin and talin. The extent of focal adhesion formation correlated with the extent of secretion and the time course of secretion also correlated with that of the assembly of focal adhesions. To examine the mechanism by which focal adhesion formation occurred, the protein kinase C inhibitor bisindolylmaleimide was used. Bisin-dolylmaleimide caused complete inhibition of both secretion and focal adhesion formation induced by antigen or the calcium ionophore A23187. Although PMA did not induce secretion, it induced focal adhesion assembly which was inhibited by bisindolylmaleimide. The inhibitor had no effect on secretion or focal adhesion formation induced by the ATP analogue, ATPγS in permeabilized cells, indicating ATPγS acts after the activation of protein kinase C in the secretory pathway. These data provide novel evidence that the formation of focal adhesions may have a role in the process of secretion from mast cells.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970310305
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  • 5
    Electronic Resource
    Electronic Resource
    Dynein family of motor proteins: Present status and future questions (1995)
    New York, NY : Wiley-Blackwell
    Cell Motility and the Cytoskeleton 32 (1995), S. 136-144 
    Details
    ISSN: 0886-1544
    Keywords: DYH1B ; dynein family ; motor proteins ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Analysis of sequence relationships in dynein heavy chains shows that dynein motor proteins comprise a single homologous family with three main branches, cytoplasmic dynein, axonemal dynein, and a third branch represented by DYH1B that lies between the other two. In all branches of the family the dynein heavy chain has four copies of the P-loop motif for a nucleotide-binding site spaced ∼300 residues apart in its midregion, with the amino acid sequence GPAGTGKT in the P-loop of the hydrolytic ATP-binding site. Cytoplasmic dyneins appear more primitive in that the heavy chain usually occurs as a homodimer, with traces of the early evolution of its four P-loop motifs by gene duplication being recognizable. In the axonemal subfamily the heavy chains occur as heterodimers or heterotrimers encoded by multiple genes, and their non-hydrolytic P-loop motifs are much more divergent with little trace of their origin by gene duplication. The DYH1B subfamily is more closely related to the cytoplasmic dyneins in sequence, but appears related to axonemal dyneins in function since it becomes upregulated during reciliation and has not been found in organisms, such as yeast and Dictyostelium, that are totally without cilia or flagella.
    Additional Material: 6 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/cm.970320214
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  • 6
    Electronic Resource
    Electronic Resource
    Laminin decreases PRL and IGFBP-1 expression during in vitro decidualization of human endometrial stromal cells (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 163 (1995), S. 30-37 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The expression of laminin, a major constituent of endometrial cell basement membranes, is increased during differentiation of human endometrial stromal cells (decidualization). To determine whether laminin plays a role in decidualization, we studied the effects of laminin substrate on the synthesis and release of prolactin (PRL) and insulin-like growth factor binding protein-1 (IGFBP-1), two major secretory proteins of decidualized stromal cells. Endometrial stromal cells were plated on laminin as well as several other extracellular matrix (ECM) proteins (types 1 and IV collagen or fibronectin) and on plastic, and cultured in media containing medroxyprogesterone acetate (MPA) and estradiol. Cells cultured on plastic or ECM proteins displayed similar morphological changes indicative of decidualization. However, the release of PRL and IGFBP-1 from cells cultured on plastic and ECM proteins (types 1 and IV collagen and fibronection) was approximately 2.1-fold and 2.8-fold greater respectively, than from cells cultured on laminin. The decrease in PRL and IGFBP-1 expression in cells cultured on laminin was not due to differences in initial cell attachment efficiency or final DNA content. In addition, laminin had no effect on the content of laminin protein or fibronectin mRNA levels, indicating that the effects of laminin on PRL and IGFBP-1 were specific. PGE2 stimulated the release of PRL and IGFBP-1 from cells cultured on laminin to levels comparable to those from cells cultured on plastic or other ECM proteins. This indicates that the decrease in PRL and IGFBP-1 release by laminin was not due to a generalized unresponsiveness. In contrast to the effects of laminin during decidualization, PRL expression was not altered by laminin in terminally differentiated decidual cells isolated at term. Our results support a role for laminin in selectively regulating PRL and IGFBP-1 gene expression during in vitro decidualization of human endometrial stromal cells. © 1995 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041630105
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  • 7
    Electronic Resource
    Electronic Resource
    Insulin-like growth factor-1 stimulates amino acid uptake by the cultured human placental trophoblast (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 165 (1995), S. 83-88 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Amino acid uptake by the human placenta is known to occur via several transport mechanisms. However, regulation by extracellular factors has received relatively little attention. A recent report by this laboratory characterized the uptake of α-aminoisobutyric acid (AIB) stimulated by insulin in the cultured human placental trophoblast The current study evaluated the effect of insulin-like growth factor-1 (IGF-1) on AIB uptake in cultured human placental trophoblasts. Na+-dependent AIB uptake was significantly stimulated by IGF-l in a time-dependent manner, as early as 30 min after hormone exposure. The maximum effect was at 2-4 hr of continuous exposure to IGF-l and the stimulation was dependent upon IGF-1 concentration approaching maximal stimulation at 50 ng.ml-1. AIB uptake was inhibited by increasing concentrations of α-(methylamino)isobtyric acid (MeAIB). Approximately 75% of basal (unstimulated) Na+-dependent AIB uptake was inhibited by MeAIB. The IGF-1-stimulated increment above basal AIB uptake was completely inhibited by MeAIB. IGF-1 increased the maximum uptake yelocity but not Km. Using equimolar concentrations, stimulation was greater with IGF-1 then with IGF-2. Stimulation by IGF-1, but not insulin, was inhibited by anit-IGF-1 receptor antibody, indicating mediation via the IGF-1 receptor. H7, a nonspecific inhibitor of serine-threonine kinase, inhibited IGF-1-dependent stimulation of AIB uptake. In addition, calphostin C (a specific inhibitor of protein kinase C), but not H89 (a specific inhibitor of protein kinase A), inhibited the IGF-1 action. This study further characterizes regulated amino acid uptake by the human placental trophoblasts and demonstrates that the Na+-dependent component of AIB uptake is stimulated by physiologic concentrations of IGF-1. © 1995 Wiley-Liss Inc.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041650111
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  • 8
    Electronic Resource
    Electronic Resource
    Induction of bovine bronchial epithelial cell filopodia by tetradecanoyl phorbol acetate, calcium ionophore, and lysophosphatidic acid (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 164 (1995), S. 123-131 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The morphological responses of primary bovine bronchial epithelial cells (BBECs) cultured in serum-free medium to protein kinase activators have been examined. When attached to type I collagen-coated tissue culture dishes, the cells responded to tetradecanoyl phorbol acetate (TPA), calcium ionophore A23187, and lysophosphatidic acid (LPA) by extruding filopodia. In contrast, no morphological changes were elicited by exposures to either epinephrine or dibutyryl-cAMP. Formation of filopodia was accompanied by actin filament reorganization as demonstrated by staining with labeled phalloidin. Exposures to varied TPA concentrations for 2 h showed maximal stimulation of filopodial extrusions at 10 nM TPA with half-maximal stimulation at 1 nM. Time-course measurements with 10 nM TPA showed filopodia formation within 30 min of exposure, with 85% of the BBECs being filopodia positive after 5 h. Filopodia induction in 20-30% of the cells could be achieved by 1-100 m̈M LPA concentrations. BBECs acquired increasing resistance to TPA-induced filopodia during the initial 5 days in culture; however, responsiveness to TPA was regenerated by mild treatment with trypsin. Inclusion of fibronectin or vitronectin into the attachment matrix had no effects on the rates or extent of TPA-induced filopodia formation. © 1995 Wiley-Liss, Inc.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041640116
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  • 9
    Electronic Resource
    Electronic Resource
    Retinoic acid nuclear receptor β inhibits breast carcinoma anchorage independent growth (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 165 (1995), S. 449-458 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Retinoids modulate cellular proliferation and mediate gene function through a series of nuclear receptors. The retinoic acid nuclear receptor β (RARβ) plays an important role in the differentiation of a number of cell types. We now demonstrate that RARβ expresion is confined to normal mammary tissue and is not expressed in either immortalized normal or malignant cell lines. Treatment of RARβ-transfected MDA-MB-231 cells with 1 μM all-trans-retinoic acid (RA) significantly inhibited monolayer growth of the cells which express recombinant RARβ. RARβ-expressing MDA-MB-231 cells formed significantly smaller and fewer colonies soft agar than the mock-transfected cells. Addition of 1 μM RA stimulated colony size and number in the RARβ-transfected MDA-MB-231 cells. In contast to the RARβ-expressing cells, colony formation by the RARβ-expressing cells was similar to the mock-transfected controls and the addition of 1 μM RA to the RARα-transfected cells inhibited colony formation. While demonstrating decreased colony formation in agar, RARβ-expressing MDA-MB-231 cells failed to exhibit decreased growth in SCID mice. Our results show that RARβ functions as a negative regulator of growth in breast epithelial cells. In addition, the growth of these cells is differentially regulated by RARα and RARβ which is most likely the result to the modulation of different genes. © 1995 Wiley-Liss Inc.
    Additional Material: 6 Ill.
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    URL: http://dx.doi.org/10.1002/jcp.1041650302
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  • 10
    Electronic Resource
    Electronic Resource
    Growth control and gene expression in a new hepatocellular carcinoma cell line, Hep40: Inhibitory actions of vitamin K (1995)
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 165 (1995), S. 459-467 
    Details
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The growth characteristics of a newly established cell line, Hep40, derived from a human hepatoma are described. An absolute requirement was found for serum to mediate cell growth. Neither EGF, TGF-α, nor HGF altered cell growth in the presence or absence of serum. A partial suppression of cell growth was achieved by several TGF-β family proteins. Affinity crosslinking gels using 125I-labeled TGF-β showed a significant decrease in the TGF-β cell-surface type II receptor in Hep40 cells, compared to the TGF-β-sensitive Hep3B cell line. However, growth could be completely suppressed by addition of vitamins K to the culture medium in both Hep40 and several other hepatoma cell lines. Growth suppression by vitamins K was accompanied by an increased level of transcripts for c-myc, c-jun, and prothrombin genes, in contrast to the actions of TGF-β1 protein, which caused a decrease in the level of c-myc transcripts. These data show that this new human hepatoma cell line has partial resistance to growth inhibition by TGF-β with a unique TGF-β receptor defect. However, growth was completely suppressed by vitamins K. The differing gene expression patterns in response to TGF-β as compared to vitamin K suggest that these two growth inhibitors act through differing pathways. © 1995 Wiley-Liss Inc.
    Additional Material: 11 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jcp.1041650303
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