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  • Life and Medical Sciences  (25)
  • Wiley-Blackwell  (25)
  • Institute of Physics
  • Wiley
  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 86 (1975), S. 143-154 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The addition of the fluorescent dye, ANS, to intact ascites tumor cells results in an enhancement of fluorescence intensity. The increase in fluorescence intensity as a function of time is biphasic which suggests that at least two processes occur. The first associated with the rapid initial rise in fluorescence represents binding to the cell surface while the second or slower phase reflects entrance of ANS into the intracellular phase. The relationship between bound and free ANS in 0.50 mM sulfate medium was used to calculated the apparent dissociation constant of ANS-membrane complex (Kd = 6.53 × 10-5 M) and the total number of ANS binding sites (4.49 nmoles/mg dry weight).Kinetic analysis of steady state sulfate transport in the presence and absence of ANS suggests that (1) sulfate exchange can be described by Michaelis Menten type kinetics (Km = 2.05 × 10-3 M), (2) a small fraction of surface associated ANS competitively inhibits sulfate exchange (Ki = 4.28 × 10-6 M) and (3) the transport system has a higher affinity for ANS than for sulfate.These data are consistent with the hypothesis that inhibition of sulfate exchange is related to the direct, reversible interaction of the negatively charged sulfonate group of ANS with superficial positively charged membrane sites.
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  • 2
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 72 (1968), S. 73-75 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Ouabain induced inhibition of cation transport and cell division in Ehrlich mouse ascites tumor cells is reversible, suggesting that this agent does not bind irreversibly to its site of action.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 78 (1971), S. 257-264 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Data from isotopic uptake experiments were used to measure the kinetics of labelling of cellular phosphate, ATP and ADP in the Ehrlich ascites tumor cell. The results show that steady state phosphate exchange flux was 0.333 ± 0.052 (S.E.) μmoles per 107 cells per hour at 37°, and that the specific activity of phosphate was the same as Pγ ATP. Metabolic inhibition reduced the phosphate flux by 30-50%. A model, based on oxidative phosphorylation and the adenylate kinase reaction is used to interpret the labelling sequence of Pβ ATP and Pβ ADP, and its dependence on Pγ ATP.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 75 (1970), S. 231-240 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Data from isotopic uptake experiments were used to measure calcium fluxes and compartment sizes in ascites tumor cells. The data were analyzed with two kinetic models, A and B. In 80% of the experiments model A, which is based on one exchangeable calcium compartment, was rejected in favor of Model B, which predicts two exchangeable compartments. A statistical evaluation of the model's performance, when fit to the experimental data was used to select between the two models.The results show that calcium was distributed between three cellular compartments in the ratio, non-exchangeable (88%): rapidly exchanging (7%): slowly exchanging (5%). The undirectional fluxes suggested that calcium transport could be described as a series system with the temporal sequence: environment ⇄ rapidly exchanging ⇄ slowly exchanging.
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  • 5
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 82 (1973), S. 435-444 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We have investigated the effects of the amino reactive reagent, 2,4,6-trinitrobenzene sulfonic acid (TNBS) on anion transport (chloride and sulfate) and on the K+ content of Ehrlich ascites tumor cells. Incubation of tumor cells with TNBS (3 mM or 10 mM) results in a time dependent uptake of this molecule. Tightly bound TNBS caused a loss of K+ as well as inhibition of sulfate uptake. Although sulfate transport was inhibited by tightly bound TNBS (40% inhibition with 20 nmoles bound per 107 cells), reversibly bound TNBS exerted much greater inhibition. Kinetic analysis of sulfate transport in the presence and absence of TNBS suggests that: (1) tightly bound TNBS exerts a competitive inhibition by occupying membrane sites remote from the specific transport site, (2) TNBS reversibly interacts with a separate site also in a competitive fashion.Increasing amounts of tightly bound TNBS resulted in an enhanced chloride influx. However, reversibly bound TNBS was without effect. These results are in contrast to the effect of TNBS on sulfate transport and show that TNBS, at least in this cell type, is not a general inhibitor of anion transport.
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  • 6
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 85 (1975), S. 1-13 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The applicability of the membrane fixed charge hypothesis to anion transport in Ehrlich ascites tumor cells was studied by investigating the dependence of steady state sulfate transport on the extracellular pH, chloride and sulfate concentration. When the extracellular sulfate was maintained at 10 mM both cellular sulfate and sulfate transport increased with decreasing pH and chloride concentration. The dependence of sulfate transport on the cellular sulfate concentration suggests a saturation phenomenon.The relationship between sulfate transport and cellular sulfate was also studied as a function of extracellular sulfate, both in the presence and absence of chloride. In both cases, sulfate transport is a saturable function of the cellular sulfate. However, in the presence of chloride the maximal flux is twice that in its absence. The discrepancy between the maximal fluxes suggests that the transport system mediates chloride-sulfate exchange in addition to sulfate self exchange. Unidirectional sulfate effluxes into chloride and sulfate-free medium; into 50 mM sulfate medium or 50 mM chloride medium were: 0.38, 1.95 and 3.91 nmoles/107 cells min-1, respectively. These results indicate that in the absence of either sulfate or chloride the net efflux, of sulfate is low. However, chloride or sulfate on the trans side of the membrane is effective in accelerating unidirectional sulfate efflux. Taken together, the results of this investigation cannot be explained in terms of the membrane fixed charge hypothesis. Rather, they support the contention that sulfate transport across the tumor cell membrane is a carrier-mediated process.
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  • 7
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 89 (1976), S. 303-311 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effects of the nonpenetrating amino reactive reagent 4-acetamido-4′-isothiocyano-stilbene-2-2′-dilsulfonic acid (SITS) on anion transport (sulfate, chloride, and inorganic phosphate) were investigated in Ehrlich ascites tumor cells. Short time exposure to SITS produces a reversible inhibition (92%) of sulfate transport. The kinetics of interaction suggest that reversibly bound SITS competitively inhibits sulfate transport, Ki = 3 × 10-6 M. Incubation of tumor cells with SITS (1 × 10-4 M) for longer periods of time results in a time dependent irreversible inhibition of sulfate transport which obeys first order kinetics. The rate coefficient for the inactivation process is 0.040 min-1. The kinetics of irreversible inhibition is best explained by the irreversible binding of SITS to the sulfate transport site, and therefore makes SITS a potentially useful probe for the quantitation of these sites in the tumor cell. The lack of effect of irreversibly bound SITS on either chloride or inorganic phosphate transport points to a specificity in the interaction of SITS with the tumor cell membrane, as well as indicating that an alternate pathway exists for the movement of these anions across the membrane.
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  • 8
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 88 (1976), S. 181-192 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The steady state transport and distribution of chloride between the intracellular and extracellular phases was investigated when the extracellular chloride concentration was varied by isosmotic replacement with nitrate, bromide and acetate. The results of these experiments show that chloride transport, measured by uptake of 36Cl, is sensitive to the replacement anion. In the presence of nitrate, chloride transport is a linear function of the extracellular chloride concentration. The relationship between chloride transport and extracellular chloride in the presence of bromide is concave upward which suggests that this anion inhibits chloride movement. However, when acetate replaces chloride, the relationship between chloride transport and extracellular chloride is concave downward. The chloride distribution ratio of cells incubated in 145-155 mM chloride medium is 0.386 and is not effected by the replacement of chloride with nitrate, bromide or acetate.These findings are consistent with the assertion that chloride transport is composed of two parallel pathways, a diffusional plus a saturating, mediated component. Of the total chloride flux (9.1 mmoles Cl-/kg dry weight per minute) measured in chloride medium (145-155 mM Cl -), the mediated component represents 40% and the diffusional component 60%.
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  • 9
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 110 (1982), S. 149-154 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The effects of extracellular Pi and Na+ on cellular Pi concentration and transport were studied. Steady-state Pi exchange flux was measured by 32P uptake in the presence and absence of Na+. Model experiments were also conducted to assess the possibility that hydrolysis of organic phosphate esters contributes to the chemically measured intracellular Pi concentration of Ehrlich ascites tumor cells. The results of these experiments indicate that hydroloysis of labile organic phosphate esters does not contribute to the measured intracellular pool of Pi. The Pi transport system exhibits an apparent Ks of 0.115 mM Pi and a maximal flux of 1.73 mmole min-1 (kg dry wt)-1. When incubated in a phosphate-buffered choline chloride medium (5 mM Pi) the intracellular Pi and the Pi influx fall by 65 and 88%, respectively. At 5 mM extracellular Pi, the Na+-dependent component of Pi transport fits Michaelis-Menten kinetics with the maximal flux equal to 2.46 mmole min-1 (kg dry wt)-1 and an apparent Ks of 35.4 mM Na+. In addition, a Na+-independent component of Pi transport, comprising about 12% of the total Pi flux, was identified. The data support the hypothesis that a Pi transport system, dependent on Na+, plays a principal role in the maintenance of intracellular Pi concentration.
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  • 10
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 121 (1984), S. 442-448 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Previous studies have shown that mediated Cl- transport which occurs by at least two processes (Cl--dependent cation cotransport and Cl- self-exchange) becomes progressively inhibited when extracellular Cl- exceeds about 60 mM (Hoffmann et al., 1979). To account for this type of kinetic behavior, that is, self-inhibition, an anion transport system possessing two sites, a high affinity transport site and a lower affinity modifier site is suggested (Dalmark, 1976). In the present experiments we have attempted to determine which of the mediated transport pathways is susceptible to self-inhibition by studying the dependence of the steady state Cl- flux on the extracellular Cl- concentration and how DIDS, an inhibitor of Cl- self-exchange, and H+ affect this relationship. Addition of DIDS to Ehrlich cells results in inhibition of Cl- transport at every Cl- concentration tested (40-150 mM). Moreover, the Cl- flux/Cl- concentration relationship no longer exhibits self-inhibition, suggesting that this phenomenon is a characteristic of the Cl- self-exchanger rather than of the Cl--dependent cation cotransport system. Lowering the extracellular pH (pHo) from 7.35 to 5.30 stimulates Cl- transport by a process that saturates with respect to [H+]. Half-maximal stimulation occurs at pHo 6.34. A comparison of the kinetic parameters, Ks and Jmax, calculated from the ascending limb of the Cl- flux/Cl- concentration curve at pHo 7.30 to those at pHo 5.50 show that the values for Ks are almost identical (23.6 mM and 21.3 mM, respectively), while the values for Jmax [(22.2 mEq/Kg dry wt). min] differ by only 15%. This finding along with the observation that DIDS completely blocks H+ stimulation of Cl- transport is compatible with the suggestion that H+ interact at the modifer site of the Cl- self-exchanger and thereby prevents self-inhibition.
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