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  • 1
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] NUCLEAR or chromosomal damage may be a major factor in radiation killing of mammalian cells1–3. The contribution of cytoplasmic effects, either directly or by induced nuclear effects, is not clear. It is known that the incorporation of 5-bromouridine deoxyriboside (BUdR) into the chromosomal ...
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  • 2
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 194 (1962), S. 660-662 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] MANY sigmoid or multi-hit curves representing the survival of cells exposed to radiation have appeared in the literature. Examples are those of Puck1 and Elkind and Sutton2, showing curves of the type described by the equation: S I - (I -(1) where S is the fraction of the population surviving a ...
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  • 3
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 335 (1980), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 122 (1985), S. 387-396 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Heat-induced alterations in CHO-10B cell Hoechst 33342 (Ho342) permeability in vitro were analyzed by flow cytometry. Immediately after 45.5°C heating, uptake was decreased in a dose-dependent manner with cytotoxicity. Kinetic analysis indicated that a treatment that reduced cell survival to approximately 10%, reduced the maximal velocity, Vmax, to 53% of control and increased the dissociation constant, Km, to 156% of control. Also, little change in Ho342 efflux was found to occur from control up to 90 min after heating. Upon incubation at 37°C after the heat treatment from 1 to 24 hr (depending on the severity of the dose) diffuse heterogeneity of Ho342 staining developed which was not evident immediately after heating. The altered staining was not due to the presence of trypan blue staining cells. Membrane permeabilization and nuclei isolation studies indicated that the lesion responsible was most likely a plasma membrane event. Induction of the heterogenous staining was not inhibited by either actinomycin D or hydroxyurea but was inhibited by incubation at 4°C. An inverse correlation existed between Ho342 permeability and clonogenicity, with nearly a 10-fold difference in survival between the high and low fluorescence intensity sorted cells. Also, larger fractions of heatsensitive S and G2M-phase cells were found in the highly fluorescent sorted fractions. These results are discussed in terms of the putative molecular events that may be involved in hyperthermic modulation of Ho342 permeability.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 135 (1988), S. 397-406 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: When CHO cells were treated either for 10 min at 45-45.5°C or for 1 hr with 100 μM sodium arsenite (ARS) or for 2 hr with 20 μg/ml puromycin (PUR-20), they became thermotolerant to a heat treatment at 45-45.5°C administered 4-14 hr later, with thermotolerance ratios at 10-3 isosurvival of 4-6, 2-3.2, and 1.7, respectively. These treatments caused an increase in synthesis of HSP families (70, 87, and 110 kDa) relative to total protein synthesis. However, for a given amount of thermotolerance, the ARS and PUR-20 treatments induced 4 times more synthesis than the heat treatment. This decreased effectiveness of the ARS treatment may occur because ARS has been reported to stimulate minimal redistribution of HSP-70 to the nucleus and nucleolus. Inhibiting protein synthesis with cycloheximide (CHM, 10 μg/ml) or PUR (100 μg/ml) after the initial treatments greatly inhibited thermotolerance to 45-45.5°C in all cases. However, for a challenge at 43°C, thermotolerance was inhibited only for the ARS and PUR-20 treatments. CHM did not suppress heat-induced thermotolerance to 43°C, which was the same as heat protection observed when CHM was added before and during heating at 43°C without the initial heat treatment. These differences between the initial treatments and between 43 and 45°C may possibly be explained by reports that show that heat causes more redistribution of HSP-70 to the nucleus and nucleolus than ARS and that redistribution of HSP-70 can occur during heating at 42°C with or without the presence of CHM. Heating cells at 43°C for 5 hr after thermotolerance had developed induced additional thermotolerance, as measured with a challenge at 45°C immediately after heating at 43°C. Compared to the nonthermotolerant cells, thermotolerance ratios were 10 for the ARS treatment and 8.5 for the initial heat treatment. Adding CHM (10 μg/ml) or PUR (100 μg/ml) to inhibit protein synthesis during heating at 43°C did not greatly reduce this additional thermotolerance. If, however, protein synthesis was inhibited between the initial heat treatment and heating at 43°C, protein synthesis was required during 43°C for the development of additional thermotolerance to 45°C. These data suggest that if a considerable amount of synthesis of HSP families occurred after the initial treatment before heating at 43°C, redistribution during 43°C of the previously synthesized HSP families could lead to the additional thermotolerance to 45°C.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 131 (1987), S. 267-275 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: When either plateau-phase or exponentially growing Chinese hamster ovary (CHO) cells are incubated in amino acid-free medium, the cells become sensitized to killing by heat. For cells deprived of amino acids for 12 h survival decreases from 1 × 10-2 for controls to 1 × 10-6 for the deprived cells, following heating at 45°C for 38 min. The survival of these sensitized cells is rapidly increased by the addition of a single amino acid just prior to heating. Of the 21 amino acids which are added in purified form to make McCoy's 5a medium, 12 show no protective effect, four have a small protective effect, and either alanine, asparagine, glutamine, serine, or theronine raise survival to a level similar to that of the control cells. The nonmetabolizable alanine analogue, 2-aminoisobutyric acid (AIB), increases survival of amino acid-deprived cells as effectively as each member of the group of five listed above, suggesting that metabolic conversion of the amino acids is not required for their protective effect. The data suggest that an increase in the intracellular concentrations of specific amino acids, independent of any change in cellular ATP content or the rate of protein synthesis, enables these cells to become quickly more resistant to killing by heat. We also conclude that the amino acid concentrations in poorly vascularized regions of some tumors should be considered, along with the oxygen, glucose, and proton concentrations, as factors which determine cellular survival following hyperthermia.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 132 (1987), S. 1-11 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: During 4 hr after puromycin (PUR: 20 μg/ml) treatment, the synthesis of three major heat shock protein families (HSPs: Mr = 110,000, 87,000, and 70,000) was enhanced 1.5-fold relative to that of untreated cells, as studied by one-dimensional gel electrophoresis. The increase of unique HSPs, if studied with two dimensional gels, would probably be much greater. In parallel, thermotolerance was observed at 10-3 isosurvival as a thermotolerance ratio (TTR) of either 2 or greater than 5 after heating at either 45.5°C or 43°C, respectively. However, thermotolerance was induced by only intermediate concentrations (3-30 μg/ml) of puromycin that inhibited protein synthesis by 15-80%; a high concentration of PUR (100 μg/ml) that inhibited protein synthesis by 95% did not induce either HSPs or thermotolerance. Also, thermotolerance was never induced by any concentration (0.01-10 μg/ml) of cycloheximide that inhibited protein synthesis by 5-94%. Furthermore, after PUR (20 μg/ml) treatment, the addition of cycloheximide (CHM: 10 μg/ml), at a concentration that reduces protein synthesis by 94%, inhibited both thermotolerance and synthesis of HSP families. Thus, thermotolerance induced by intermediate concentrations of PUR correlated with an increase in newly synthesized HSP families.This thermotolerance phenomenon was compared with another phenomenon termed heat resistance and observed when cells were heated at 43°C in the presence of CHM or PUR immediately after a 2-hr pretreatment with CHM or PUR. Heat protection increased with inhibition of synthesis of both total protein and HSP families. Moreover, this heat protection decayed rapidly as the interval between pretreatment and heating increased to 1-2 hr, and did not have any obvious relationship to the synthesis of HSP families. Therefore, there are two distinctly different pathways for developing thermal resistance. The first is thermotolerance after intermediate concentrations of PUR treatment, and it requires incubation after treatment and apparently the synthesis of HSP families. The second is resistance to heat after CHM or PUR treatment immediately before and during heating at 43°C, and it apparently does not require synthesis of HSP families. This second pathway not requiring the synthesis of HSP families also was observed by the increase in thermotolerance at 45.5°C caused by heating at 43°C after cells were incubated for 2-4 hr following pretreatment with an intermediate concentration of PUR.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 132 (1987), S. 41-48 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: After sodium arsenite (100 μM) treatment, the synthesis of three major heat shock protein families (HSPs; Mr = 110,000, 87,000, and 70,000), as studied with one-dimensional gels, was enhanced twofold relative to that of unheated cells. The increase of unique HSPs, if studied with two-dimensional gels, would probably be much greater. In parallel, thermotolerance was observed as a 100,000-fold increase in survival from 10-6 to 10-1 after 4 hr at 43°C, and as a thermotolerance ratio (TTR) of 2-3 at 10-3 isosurvival for heating at 45.5°C. Cycloheximide (CHM: 10 μg/ml) or puromycin (PUR: 100 μg/ml), which inhibited total protein synthesis and HSP synthesis by 95%, completely suppressed the development of thermotolerance when either drug was added after sodium arsenite treatment and removed prior to the subsequent heat treatment. Therefore, thermotolerance induced by arsenite treatment correlated with an increase in newly synthesized HSPs. However, with or without arsenite treatment, CHM or PUR added 2-6 hr before heating and left on during heating caused a 10,000-100,000-fold enhancement of survival when cells were heated at 43°C for 4 hr, even though very little synthesis of heat shock proteins occurred. Moreover, these cells manifesting resistance to heating at 43°C after CHM treatment were much different than those manifesting resistance to 43°C after arsenite treatment. Arsenite-treated cells showed a great deal of thermotolerance (TTR of about 10) when they were heated at 45°C after 5 hr of heating at 43°C, compared with less thermotolerance (TTR of about 2) for the CHM-treated cells heated at 45°C after 5 hr of heating at 43°C. Therefore, there are two different phenomena. The first is thermotolerance after arsenite treatment (observed at 43°C or 45.5°C) that apparently requires synthesis of HSPs. The second is resistance to heat after CHM or PUR treatment before and during heating (observed at 43°C with little resistance at 45.5°C) that apparently does not require synthesis of HSPs. This phenomenon not requiring the synthesis of HSPs also was observed by the large increase in thermotolerance to 45°C caused by heating at 43°C, with or without CHM, after cells were incubated for 6 hr following arsenite pretreatment. For both phenomena, a model based on synthesis and redistribution of HSPs is presented.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 163 (1995), S. 194-203 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: To study the mechanism of thermotolerance, the adaptive response by which cells become transiently resistant to killing by heat shock, we have focused on the centrosome, an organelle whose disorganization is closely correlated with thermal killing in Chinese hamster ovary (CHO) cells. Centrosome structure was studied by use of antisera directed against pericentrin, a 220 Kd protein of the pericentriolar material (PCM). Centrosome function was measured in intact cells by performing microtubule regrowth following exposure to the drug nocodazole. Immediately following heating at 45°C for 4-18 min, centrosomal staining by antipericentrin decreased. Thereafter, staining gradually recovered, although abnormal configurations of staining appeared in heated cultures 10-20 h later. In contrast, abnormal patterns of staining rarely developed in thermotolerant cultures. Centriole number was not perturbed by heat, indicating that the heat effect was specific for the PCM. Heat also caused an immediate reduction in the number of microtubules nucleated by the PCM. As for staining by antipericentrin, microtubule nucleation recovered during 3-20 h at 37°C after heating. The immediate, heat-induced decrease in antipericentrin staining or microtubule nucleation was similar in thermotolerant and nontolerant cells. In contrast, the inhibition for both endpoints recovered to control levels much more quickly in thermotolerant cells than in nontolerant cells. Furthermore, new protein synthesis was not required for the recovery of microtubule nucleation. These data show that thermotolerant cells have an enhanced capacity to repair thermal damage to centrosome structure and function, and suggest that a faster rate of recovery prevents disorganization of the PCM that is observed in nontolerant cells several hours after heating. © 1995 Wiley-Liss, Inc.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 140 (1989), S. 227-232 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The temperature-sensitive (ts) Chinese hamster ovary (CHO) cell mutant tsH1 contains a thermolabile leucyl-tRNA synthetase. Upon incubation at the nonpermissive temperature of 39.5°C, the enzyme became reversibly inhibited over a period of minutes, and the cells lost viability over a period of many hours. However, killing of tsH1 by acute heating at 45°C was identical to that of wild-type (SC) cells. In addition, the heat-induced inhibition of protein synthesis was similar for both cell types, as measured after acute heating at 45°C. Furthermore, both killing and inhibition of protein synthesis showed thermotolerance in both cell types. In contrast to the effects at 45°C, at 39.5°C, neither the inhibition of leucyltRNA synthetase activity nor the killing of tsH1 expressed thermotolerance. Also, treatment of tsH1 at 39.5°C did not induce thermotolerance to killing at 45°C. The inhibition of leucyl-tRNA synthetase activity in tsH1 at 39.5°C was further distinguished from the 45°C-induced inhibition of protein synthesis in SC cells by a much more rapid reversal of the inhibition of leucyl-tRNA synthetase activity. Also, the rate of reversal of the inhibition of protein synthesis by 45°C in SC cells was decreased by increased heat dose. Such was not true for the 39.5°C inhibition of leucyl-tRNA synthetase activity in tsH1. The data indicate that there exist two distinct types of thermal inhibition-one slowly reversible type which was observed during and after heating at 45°C and both induced and expressed thermotolerance, and a second, rapidly reversible type, which was evident only during heating of tsH1 at 39.5°C and neither induced nor expressed thermotolerance.
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