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  • fluorescence decay  (3)
  • Anaerobiosis  (2)
  • DNA synthesis  (2)
  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Journal of cancer research and clinical oncology 95 (1979), S. 115-122 
    ISSN: 1432-1335
    Keywords: Acetat ; Anaerobiose ; Lipogenese ; Oktanoat ; Pyruvate ; Tritiiertes Wasser ; Tumor ; Acetate ; Anaerobiosis ; Lipogenesis ; Octanoate ; Pyruvate ; Tritiated water ; Tumor
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Anaerobic culture conditions (95% argon/5% CO2) caused a slightly greater increase in total lipids of Ehrlich ascites tumor cells than a gas phase of 20% O2, 75% N2, 5% CO2. Whereas the rate of [U-14C]acetate incorporation into total lipids and lipid-subclasses rose markedly in the absence of oxygen, a drastic decrease of [U-14C]pyruvate and [1-14C]octanoate incorporation as well as a 30% reduction of 3H incorporation into lipids from tritiated water were observed under these conditions. Since profound changes in the metabolic state of cells cause alterations in the specific activity of the acetyl-CoA pool but do not alter the specific activity of intracellular water, this precursor is considered to be an adequate monitor for lipogenesis under aerobic and anaerobic culture conditions. Therefore, it is concluded that Ehrlich ascites tumor cells are not able to reoxidize NADH/NADPH in the absence of oxygen by a stimulation of biosynthesis of fatty acids as is discussed to be the case in normal cells. The slight increase in total lipids of anaerobically cultured cells seems to be the result of an imbalance between normal uptake and impaired utilization of lipids from serum-supplemented culture medium.
    Notes: Zusammenfassung Unter anaeroben Kulturbedingungen, (95% Argon 5% CO2) stieg der Gesamtlipidgehalt in Ehrlich Ascites Tumorzellen etwas stärker an als unter aeroben Bedingungen (20% O2, 75% N2, 5% CO2). In Abwesenheit von Sauerstoff war der Einbau von [U-14C]Acetat in die Gesamtlipidfraktion als auch in einzelne Lipidklassen, deutlich erhöht, wohingegen ein drastischer Abfall der Einbauraten aus [U-14C]Pyruvat und [1-14C]Oktanoat sowie eine signifikante Verminderung des Einbaus von 3H-Wasser unter diesen Kulturbedingungen beobachtet wurde. Da die spezifische Aktivität von intrazellulärem Wasser auch unter tiefgreifenden Änderungen des metabolischen Zustandes der Zellen praktisch konstant bleibt, wird 3H-Wasser als die zuverlässigste Vorstufe zur Messung der Lipidbiosynthese unter aeroben und anaeroben Kulturbedingungen angesehen. Aus den Ergebnissen wird geschlossen, daß Ehrlich Ascites Tumorzellen nicht, wie es für Normalgewebe angenommen wird, in der Lage sind, NADH/NADPH bei Ausschluß von Sauerstoff durch erhöhte Fettsäuresynthese zu, reoxidieren. Der beobachtete Anstieg im Gesamtlipidgehalt von anaerob kultivierten Zellen erklärt sich wahrscheinlich durch ein Ungleichgewicht zwischen normaler Aufnahme und gehemmtem Abbau von exogenen Lipiden.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1432-0738
    Keywords: Key words Organochlorine pesticides ; Cultured oviductal and uterine cells ; DNA synthesis ; Estrogen receptor ; Bovine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract  The pesticides DDT, MXC and γHCH at concentrations between 41 and 200 μM inhibited DNA synthesis (measured by [3H]thymidine incorporation) of cultured bovine oviductal endosalpingeal and uterine cells in the order DDT〉MXC〉γHCH, in comparison to nonexposed controls. Sensitivity to the toxicants was greater in uterine epithelial and stromal cells than in uterine smooth muscle or oviductal endosalpingeal cells. Besides the inhibitory effect, there was a stimulatory effect on DNA synthesis in epithelial cells in the range of 28 nM to 2.8 μM DDT and in stromal cells at 2.8 and 28 nM for MXC. An explanation for this reaction could be that both toxicants have an estrogen-like effect. In the present study, it is shown that the o,p’ isomer of DDT can bind to the cytoplasmatic estrogen receptor and DDT or MXC were able to inhibit the binding of radiolabelled estradiol to the uterine endometrial explants in bovine, whereas γHCH did not change the binding. These findings represent an estrogenic effect of DDT and MXC in two complete in vitro systems.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Journal of cancer research and clinical oncology 113 (1987), S. 298-300 
    ISSN: 1432-1335
    Keywords: Ehrlich ascites tumor cells ; DNA over-replication ; Anaerobiosis ; G2M phase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary G2-enriched fractions of Ehrlich ascites tumor cells (up to 80%–85% G2 cells) separated from anaerobically and aerobically cultured asynchronous populations by centrifugal elutriation revealed the same growth characteristics after recultivation under standard conditions: a significant proportion of cells with increased DNA (DNA content〉4C) emerged. Interruption of DNA synthesis by deprivation of oxygen may account for polyploidisation (over-replication) of DNA but other mechanisms must be taken into consideration.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-1335
    Keywords: Ehrlich ascites tumor cells ; Methylglyoxal ; Glucosone ; Galactosone ; Growth inhibition ; DNA synthesis ; Protein Synthesis ; Energy Metabolism
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary 1. Proliferation of in vitro grown Ehrlich ascites tumor cells is completely inhibited by 0.2–0.4 mM methylglyoxal and 1–2mM glucosone or galactosone without severely affecting viability (dye exclusion test); no phase-specific arrest of cell growth is observed. 2. Incorporation of [14C] thymidine into the acid-insoluble fraction of the cells decreases within a few minutes to less than 50% of that in controls in the presence of 0.4 mM methylglyoxal, and 2 mM glucosone or galactosone causes a comparable inhibition of DNA synthesis after 2 h or 4 h, respectively. 3. The action of 0.4 mM methylglyoxal inhibits incorporation of [14C] leucine within a few minutes by more than 70%, while 2 mM glucosone and galactosone are significantly less effective (50%–60% inhibition after 12 h). 4. While methylglyoxal and galactosone do not severely affect lactate production of the cells, 2 mM glucosone reduces glycolysis by 60%–70%; ATP/ADP ratios did not fall below 3.5 in the presence of the inhibitors (controls 4–6). 5. It is suggested that the reaction potentialities of the oxaldehyde function of the inhibitors play an important role in their growth-inhibitory acitivity, besides exerting a specific effect on hexokinase (glucosone) and UTP-trapping activity.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Journal of fluorescence 6 (1996), S. 165-168 
    ISSN: 1573-4994
    Keywords: Cation binding ; fluorescence decay ; kinetics ; binding constants ; Na,K-ATPase ; eosin Y
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract Time-resolved fluorescence and binding studies have been carried out on Na,K-ATPase in the presence of the fluorescent dye eosin Y to obtain thermodynamic and kinetic parameters for the interaction of the enzyme with different cations. Eosin Y binding is indicated by a 3 ns fluorescence decay process and is observed only in the presence of mono- and divalent cations. This type of cation binding is interpreted as a nonselective electrostatic interaction, with negatively charged groups of the enzyme providing a high-affinity eosin Y binding site. Eosin Y binding is observed only under conditions where the enzyme exists in the conformational state F1. The kinetic parameters of eosin Y binding have been determined employing stopped-flow fluorometry.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1573-4994
    Keywords: FITC ; antibodies ; fluorescence decay ; Na ; K-ATPase ; pK
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract Stationary and time-resolved fluorescence of FITC–Na,K-ATPase is investigated as a function of pH in the presence of different ligands, cations, and the monoclonal anti-FITC antibody 4-4-20. The binding of K+ and of the antibody leads to the same decreased fluorescence intensity level. Antibody binding is observed only under conditions where the enzyme exists in the conformational state F1, and not in the form of the Na+ or K+ complex or when it is phosphorylated with inorganic phosphate in the presence of Mg2+. For the interpretation of the results it is shown that the fluorophore is not essentially affected by an acidity change of the bound dye, so that pK variations responsible for the observed intensity changes can be excluded in favor of a static quenching process
    Type of Medium: Electronic Resource
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  • 7
    ISSN: 1573-4994
    Keywords: Fluorescence spectra ; fluorescence decay ; dissociation constants ; fluorescein 5′-isothiocyanate ; FITC-fluorescein 5′-isothiocyanate-Na+,K+-ATPase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Physics
    Notes: Abstract The fluorescence emission intensity between the Na+, and the K+ complex of Na+,K+-ATPase, labeled with fluorescein 5′-isothiocyanate (FITC), differs by 30 to 40%. Experimental studies are carried out to elucidate the physical reasons which account this intensity difference. The dissociation constant of protolysis of the covalently bound FITC and its fluorescence decay times are determined in media of different ionic compositions and are compared with the corresponding properties of a synthetic model compound. The fluorophore bound to the protein is characterized by two decay times in the nanosecond range; the model compound, by a single one. The static fluorescence intensity changes are discussed on the basis of these results.
    Type of Medium: Electronic Resource
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