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  • 1
    Digitale Medien
    Digitale Medien
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 215 (2002), S. 0 
    ISSN: 1574-6968
    Quelle: Blackwell Publishing Journal Backfiles 1879-2005
    Thema: Biologie
    Notizen: In this report we describe the development of a highly stringent and dually regulated promoter system for Shigella flexneri. Dual regulation was provided by utilizing a promoter susceptible to control by the bacteriophage P1 temperature-sensitive C1 repressor that in turn was under the transcriptional control of LacI. The level of induction/repression ratios observed was up to 3700-fold in S. flexneri. The general utility of this promoter system was evaluated by demonstrating that the bacteriophage P1 post-segregational killer protein Doc mediates a bactericidal effect in S. flexneri. This represents the first report of Doc (death on curing)-mediated killing in this Gram-negative species.
    Materialart: Digitale Medien
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  • 2
    Digitale Medien
    Digitale Medien
    [s.l.] : Nature Publishing Group
    Nature 248 (1974), S. 422-424 
    ISSN: 1476-4687
    Quelle: Nature Archives 1869 - 2009
    Thema: Biologie , Chemie und Pharmazie , Medizin , Allgemeine Naturwissenschaft , Physik
    Notizen: [Auszug] FIG. 1 a, Cells grown in roller culture were collected by shaking, pelleted at 8000, and washed 2 h in minimal essential medium (MEM) buffered with HEPES pH 7.6 at 4 C. Aliquots of cells in 10 ml of MEM were distributed evenly to 25 ml Erlenmeyer flasks and warmed to 37 C. 1, 2, 6, 7 ...
    Materialart: Digitale Medien
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  • 3
    Digitale Medien
    Digitale Medien
    Springer
    Molecular and cellular biochemistry 67 (1985), S. 47-53 
    ISSN: 1573-4919
    Schlagwort(e): α1-adrenergic receptor ; membrane ; photoaffinity ; smooth muscle
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Summary In this study, we have used an α1-adrenergic receptor photoaffinity ligand, 2-[4-(4-azido-3-iodo-benzoyl)-piperazin-1-yl]-4-amino-6, 7-dimethoxyquinazoline (125I-APD), to label covalently the α1-adrenergic receptor in a smooth muscle cell line. Our results indicate that in the absence of light, (125I)APD binds reversibly to a site in the DDT1 MF-2 cell membranes having pharmacological characteristics of an α1-adrenergic receptor. Following incorporation of (125I)ADP into partially purified membranes a single labeled band of protein with a Mr of 81 000 was visualized by autoradiography following sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Incorporation of (125I)-APD into this band was affected by adrenergic agonists and antagonists in a manner consistent with an α1-adrenergic interaction. Prazosin (α1-selective) blocked incorporation of the label into the Mr = 81 000 protein while yohimbine (α2-selective) did not. Of the adrenergic agonists, (−)-epinephrine and (−)-norepinephrine but not (−)-isoproterenol blocked labeling of the Mr − 81 000 protein. We conclude that the ligand binding site of the DDT1 MF-2 cell α1-adrenergic receptor resides in a Mr = 81 000 protein.
    Materialart: Digitale Medien
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  • 4
    ISSN: 1573-4919
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract We have demonstrated that glucocorticoids induce in DDT1 MF-2 cells by a glucocorticoid mediated mechanism the synthesis of a methionine-cysteine rich protein of 29 000 Mr (p29). Induction of p29 is not observed in DDT1 MF-2 GR glucocorticoid resistant variants which have only 7% of glucocorticoid receptor site per cell compared to wild type cells. Increased synthesis of p29 is specific to glucocorticoids since neither androgens, estrogens, progesterone nor the glucocorticoid antagonist dexamethasone mesylate are effective inducers. Stimulation of p29 synthesis in wild type cells is observed at 10−10 M triamcinolone acetonide, reaching a maximum at a concentration of 1 × 10−8 M. The induction of p29 is not a function of glucocorticoid arrest of DDT1 MF-2 cells since DDT1 MF-2 cells promoted to re-enter the cell cycle by 50 ng/ml platelet derived growth factor (PDGF) continue synthesis of p29. Finally, increased levels of p29 translation products are observed in cell free translation assays carried out utilizing poly A− RNA transcripts isolated from glucocorticoid treated cells. These data suggest that the glucocorticoid stimulation of p29 synthesis is a transcriptional and/or RNA processing event controlled by glucocorticoid receptor complexes.
    Materialart: Digitale Medien
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  • 5
    ISSN: 1573-4919
    Schlagwort(e): glucocorticoid receptor ; β-adrenergic receptor regulation ; smooth muscle ; adenylate cyclase
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie , Medizin
    Notizen: Abstract We have shown that glucocorticoids induce the appearance of β2-adrenergic receptors in membranes of the ductus deferens smooth muscle cell line (DDT1 MF-2). A concomitant increase in isoproterenol stimulated adenylate cyclase activity in the absence of exogenously applied GTP was observed as was a significantly increased (p 〈 0.05) sensitivity of the adenylate cyclase system to exogenously applied GTP. However, no significant difference in the maximal velocity of adenylate cyclase between control and steroid treatment was measurable in the presence of sodium fluoride. Induction of β2-adrenergic receptors in DDT1 MF-2 cells is correlated with the presence of steroid receptors (androgen and glucocorticoid) in the cells since estrogens and progesterones had no effect on receptor levels. Finally, utilizing dense amino acid labeling of cells to measure old versus newly synthesized receptor sites by a density shift method, we have documented that glucocorticoid induction of β2-adrenergic receptors involves synthesis of new receptor protein.
    Materialart: Digitale Medien
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  • 6
    ISSN: 1573-6865
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Medizin
    Notizen: Abstract Thrombospondin 1 is an extracellular matrix glycoprotein with multiple functions. In the skin, it has been immunolocalized to basement membrane, and its expression increases during embryogenesis and wound healing. Its normal cellular source in the skin is not known, except during wound healing, where macrophages and keratinocytes seem to be the primary source. We have analysed the expression of thrombospondin 1 mRNA in normal mouse skin at different ages by in situ hybridization. It was found that the mRNA is expressed by dermal mesenchymal cells and mature fibroblasts and developmentally regulated during post-natal skin growth and morphogenesis. In adult mouse skin, expression of the thrombospondin is restricted to the mesenchymal cells of hair follicle papilla. These results suggest that the regulation of thrombospondin 1 transcription in mesenchymal cells can play an important role in post-natal skin development. Its mRNA expression is a characteristic of a dult dermal papilla cells with a potential role in hair development.
    Materialart: Digitale Medien
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  • 7
    Digitale Medien
    Digitale Medien
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 156 (1993), S. 88-95 
    ISSN: 0021-9541
    Schlagwort(e): Life and Medical Sciences ; Cell & Developmental Biology
    Quelle: Wiley InterScience Backfile Collection 1832-2000
    Thema: Biologie , Medizin
    Notizen: The DDT1 MF2 smooth muscle cell line was derived from an estrogen/androgeninduced leiomyosarcoma arising in the hamster ductus deferens. Growth of this cell line is arrested in Go/G1 by treatment with glucocorticoids. To facilitate the study of the mechanism of glucocorticoid-induced cell growth arrest, a glucocorticoid-resistant variant cell line, DDT1 MF2 GR1 (GR1), was developed by genetic selection. Growth of this mutant cell line is completely resistant to the inhibitory action of glucocorticoids. However, we now demonstrate that both primary and secondary glucocorticoid-induced events still exist in the GR1 cell line. By analyzing the expression and genetic pattern of glucocorticoid receptor, no detectable rearrangement of the glucocorticoid receptor gene was found although the expression of both mRNA and protein levels of the receptor were lower in the variant compared to wild-type cells. In addition, we found that the expression of two growth-associated genes, Ha-ras and transforming growth factor β1 (TGF-β1) are down-regulated by glucocorticoids in wild-type DDT1 MF2 cells but not in GR1 cells. These results indicated that the function or activity of glucocorticoid receptor in the GR1 cells is not qualitatively altered. Our data suggest that a lower glucocorticoid receptor level is not the real cause or at least not the single cause for the GR1 cell's loss of sensitivity to the inhibitory action of glucocorticoid. Instead, we postulate the existence of a defect downstream of the primary site of action of glucocorticoid receptor complexes in GR1 cells. © 1993 Wiley-Liss, Inc.
    Zusätzliches Material: 5 Ill.
    Materialart: Digitale Medien
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