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  • Life and Medical Sciences  (57)
  • Engineering  (52)
  • 1
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 152 (1992), S. 632-638 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In cultured mammalian cells, the relationship was investigated between mitochondrial function and proliferation under various culture conditions. Continuous inhibition of the expression of the mitochondrial genome was used to reduce the activity of enzymes involved in oxidative phosphorylation by 50% at every cell division. Under these conditions, culturing in relatively poor media resulted in arrest of the proliferation of most cell lines after 1 cell division. This was preceded by decreasing levels of ATP and increasing levels of ADP, suggesting that the ATP-generating capacity of the cells was limiting. Culturing in richer media led to arrest of the proliferation after 5 to 6 divisions, but accumulation of ADP was not observed. Addition of pyruvate to rich culture media and, at least for 1 cell line, increasing the CO2 levels, completely prevented proliferation arrest. Inability to synthesise metabolic precursors via mitochondrial intermediary metabolism probably explains growth arrest of cells cultured in rich media. Pyruvate and CO2 were, however, without effect on the proliferation arrest of cells cultured in relatively poor media. Therefore, pyruvate dependency for growth of cells without functional mitochondria holds true only under culture conditions where the ATP-generating capacity of the cells is not limiting. © 1992 Wiley-Liss, Inc.
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  • 2
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A431 cells grown as three-dimensional spheroids show growth stimulation in response to nanomolar concentrations of EGF in contrast to monolayer cultures that show inhibition. In investigating the alterations in EGF signal transduction that underlie this modification of the proliferative response, we have compared the expression of EGF receptors on A431 cells under these conditions and related our findings to tyrosine phosphorylation and the growth response. EGF receptors were measured by 125I-EGF binding to trypsin-dispersed cells. Unexpectedly, dispersion of the monolayers caused an 80% decrease in surface EGF receptor, although, after dispersion, EGF receptor was digested by trypsin with a half-life of 69 ± 32 min. No evidence for a comparable loss of cellular EGF receptor was seen on trypsin dispersion of spheroids. After allowing for this effect, we found that the receptor density on nondispersed monolayers (5 × 106 per cell) was twentyfold greater than that on spheroids (0.25 × 106 per cell). EGF-induced tyrosine phosphorylation was confined to the outermost cells of the spheroid, although the presence of surface-expressed EGF pinding sites could be demonstrated throughout the structure and the number of EGF receptors/cell on dispersed spheroid cells showed a single distribution peak by flow cytometry, with no evidence for more than one population. Using RCM-lysozyme as a substrate, tyrosine phosphatase activity in spheroids lay within the range observed in monolayer cultures. Autophosphorylation of the EGF receptor following EGF stimulation in monolayer cultures of A431 cells rose rapidly in the first 10 seconds and then slowly increased for at least 3 h. In spheroids, it reached a maximum within 10 seconds and then declined over 3 h. Since the microenvironment within a tumor resembles that in a spheroid, a similar reduction in surface EGF receptor expression may be expected in tumors relative to monolayer cultures, together with corresponding growth stimulation in response to EGF. © 1994 Wiley-Liss, Inc.
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  • 3
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 161 (1994), S. 23-30 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The alpha granules of circulating platelets are dynamic structures that acquire endogenous and exogenous components by synthesis and uptake, respectively. The uptake of exogenous components is a result of either receptor-mediated endocytosis or fluid-phase pinocytosis. Despite many detailed studies on the function and content of α-granules, little is known of the impact of platelet age on these organelles. In this report, we describe the use of platelet biotinylation to identify and isolate aged platelets for the analysis of α-granule contents. When aged platelets were permeabilized and examined by flow cytometry utilizing fluorescently labeled antibodies, two exogenously acquired proteins, fibrinogen and immunoglobulin G, were found to increase significantly with platelet age. The levels of intracellular fibrinogen were found to be elevated relative to control, 114 ± 2% and 119 ± 5% on days 4 and 5 postbiotinylation, respectively; the life span of dog platelets is 6.0 days. Intracellular immunoglobulin G content increased similarly. Levels of two endogenously synthesized proteins, thrombosponding and P-selectin, were not elevated in aged platelets. Confirmation of the flow cytometric data was obtained by isolating aged, biotinylated platelets by fluorescence-activated cell sorting and quantitating the fibrinogen levels with an ELISA assay. For platelets averaging 4.6 days of age, the fibrinogen level was elevated to 128 ± 23% of the level for the entire platelet population. These data demonstrate that age-dependent changes in exogenously acquired α-granule proteins do occur and that the uptake mechanism for these proteins is active through out the platelet life span. © 1994 Wiley-Liss, Inc.
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  • 4
    Electronic Resource
    Electronic Resource
    New York, NY [u.a.] : Wiley-Blackwell
    Journal of Cellular Physiology 142 (1990), S. 334-341 
    ISSN: 0021-9541
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Epidermal melanocytes (MC) synthesise melanin in response to ultraviolet radiation (UVR). The mechanisms mediating the UV-induced activation of melano-genesis are unknown but since UVR induces turnover of membrane phospholipids generating prostaglandins (PGs) and other products, it is possible that one of these might provide the activating signal. We have examined the effects of prostaglandins (PGs) E1, E2, D2, F2α, and di-acyl glycerol upon the UV-induced responses of cultured human MC and the Cloudman S91 melanoma cell line. The PGs had little effect on unirradliated cells and did not alter the response to UVR in either human MC or S91 melanoma cells. However, a synthetic analogue of di-acyl glycerol, 1-oleyl 2-acetyl glycerol (OAG), caused a significant (P〈0.0001), dose-related augmentation of melanin content both in human MC (seven-fold) and S91 cells (three-fold). UVR caused a significant augmentation of the OAG-induced melanognesis of both human MC and S91 cells. Since OAG is known to activate protein kinase C, it was possible that the observed modulation of the UVR signal could be via that pathway. Di-octanoyl glycerol, another di-acyl glycerol, which activates kinase C, caused a small (70%) increase in melanogenesis in MC which was not altered by UVR. However, 12-0 tetradecanoyl phorbol 13-acetate (TPA), a potent activator of protein kinase C, had no significant effect on either basal or UV-induced melanin synthesis in either cell type. These data suggest that the UV-induced signal activating melanogenesis could be mediated by di-acyl glycerol. Furthermore, they imply that the signal is transduced via an alternative, pathway that might be independent of protein kinase C.
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  • 5
    Electronic Resource
    Electronic Resource
    Chichester : Wiley-Blackwell
    International Journal for Numerical Methods in Fluids 14 (1992), S. 477-493 
    ISSN: 0271-2091
    Keywords: Bed friction ; Hydrodynamics ; Shallow sea ; Galerkin ; Engineering ; Engineering General
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: The mathematical formulation of a three-dimensional shallow sea model using a modal expansion in the veitical is briefly described.The importance of the time discretization of the vertical diffusion term and bottom friction term is discussed in some detail. Both stability theory and numerical calculations show the importance of time centring or evaluating the modal form of the viscosity term at the higher time step in order to develop a numerically efficient algorithm. Similar analysis and calculations show that in shallow water it is essential to time centre or evaluate bottom friction at the higher time step. In the case of linear bottom friction it is shown that this condition can be readily accomplished. However, using a quadratic friction formulation (a more physically realistic form), this cannot be readily achieved. A new algorithm is presented whereby a stable solution can be obtained even in shallow water using quadratic bottom friction.
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  • 6
    Electronic Resource
    Electronic Resource
    Chichester : Wiley-Blackwell
    International Journal for Numerical Methods in Fluids 1 (1981), S. 3-16 
    ISSN: 0271-2091
    Keywords: Vector Upstream ; Differencing Scheme ; Significant Sources ; False Diffusion ; Spatial Oscillation ; Steady-state Linear Systems ; Convective Flow ; Rod Bundle Geometry ; Temperature Prediction ; Engineering ; Engineering General
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: This paper considers a finite difference scheme for modelling the convection/diffusion equation in strongly convective flow regimes including circumstances in which significant source terms are present.The main objective is to provide an alternative approach to central and/or upwind difference methods which for various reasons are unsatisfactory. To illustrate the main features of the scheme, an assessment of its accuracy is made by means of a Taylor expansion analysis and a study of its performance in two model problems. As a demonstration of its generality for use in large-scale practical problems, some numerical results are presented for the prediction of the temperature distribution in a flow through a partially blocked heated rod bundle.The main conclusions are that in almost all practical circumstances results obtained using the scheme are not susceptible to false diffusion or spatial oscillations, which are, respectively, the inherent weaknesses in many upwind and central difference scheme formulations, and in general its use results in improved overall accuracy.
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  • 7
    Electronic Resource
    Electronic Resource
    Chichester : Wiley-Blackwell
    International Journal for Numerical Methods in Fluids 2 (1982), S. 151-171 
    ISSN: 0271-2091
    Keywords: Penalty method ; Incompressible Flow ; Finite Elements ; Convergence ; Existence ; Engineering ; Engineering General
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: In this paper the penalty function method is reviewed in the general context of solving constrained minimization problems. Mathematical properties, such as the existence of a solution to the penalty problem and convergence of the solution of a penalty problem to the solution of the original problem, are studied for the general case. Then the results are extended to a penalty function formulation of the Stokes and Navier-Stokes equations. Conditions for the equivalence of two penalty-finite element models of fluid flow are established, and the theoretical error estimates are verified in the case of Stokes's problem.
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  • 8
    Electronic Resource
    Electronic Resource
    Chichester : Wiley-Blackwell
    International Journal for Numerical Methods in Fluids 9 (1989), S. 23-41 
    ISSN: 0271-2091
    Keywords: Momentum transport ; Turbulent mixing layers ; Discrete vortex method ; Time-dependent momentum fluctuations ; Comparison with experiments ; Large-scale structures ; Engineering ; Engineering General
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: The turbulent momentum transport phenomena in a two-dimensional mixing layer are investigated numerically by a discrete vortex method. The numerical model and calculations are verified through a comparison with existing numerical simulations and experimental measurements. The main emphasis is placed on the exploration of the detailed time-dependent instantaneous local momentum fluctuations and on the comparison of numerical results with available experimental measurements. The current simulations confirm qualitatively the various trends in the turbulent momentum flux and fluctuating components of the velocity in the mixing layer found with several experimental results. The study shows that similarity exists in turbulent momentum quantities along the axial direction of the mixing layer. The calculations also show a definite correlation between the passage of a large-scale structure and a burst in the turbulent momentum flux. The probability density functions of the fluctuating quantities are shown to be mostly Gaussian-like, with only a few exceptions.
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  • 9
    Electronic Resource
    Electronic Resource
    Chichester : Wiley-Blackwell
    International Journal for Numerical Methods in Fluids 10 (1990), S. 373-400 
    ISSN: 0271-2091
    Keywords: Comparison ; Experimental ; Simulation ; Planar entry flow ; Viscoelastic ; Convergence ; Engineering ; Engineering General
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: The goal of this research was to determine whether there is any interaction between the type of constitutive equation used and the degree of mesh refinement, as well as how the type of constitutive equation might affect the convergence and quality of the solution, for a planar 4:1 contraction in the finite eiement method. Five constitutive equations were used in this work: the Phan-Thien-Tanner (PTT), Johnson-Segalman (JS), White-Metzner (WM), Leonov-like and upper convected Maxwell (UCM) models. A penalty Galerkin finite element technique was used to solve the system of non-linear differential equations. The constitutive equations were fitted to the steady shear viscosity and normal stress data for a polystyrene melt. In general it was found that the convergence limit based on the Deborah number De and the Weissenberg number We varied from model to model and from mesh to mesh. From a practical point of view it was observed that the wall shear stress in the downstream region should also be indicated at the point where convergence is lost, since this parameter reflects the throughput conditions. Because of the dependence of convergence on the combination of mesh size and constitutive equation, predictions of the computations were compared with birefringence data obtained for the same polystyrene melt flowing through a 4:1 planar contraction. Refinement in the mesh led to better agreement between the predictions using the PTT model and flow birefringence, but the oscillations became worse in the corner region as the mesh was further refined, eventually leading to the loss of convergence of the numerical algorithm. In comparing results using different models at the same wall shear stress conditions and on the same mesh, it was found that the PTT model gave less overshoot of the stresses at the re-entrant corner. Away from the corner there were very small differences between the quality of the solutions obtained using different models. All the models predicted solutions with oscillations. However, the values of the solutions oscillated around the experimental birefringence data, even when the numerical algorithm would not converge. Whereas the stresses are predicted to oscillate, the streamlines and velocity field remained smooth. Predictions for the existence of vortices as well as for the entrance pressure loss (ΔPent) varied from model to model. The UCM and WM models predicted negative values for ΔPent.
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  • 10
    Electronic Resource
    Electronic Resource
    Chichester : Wiley-Blackwell
    International Journal for Numerical Methods in Fluids 7 (1987), S. 1325-1342 
    ISSN: 0271-2091
    Keywords: Convective Heat Transfer ; Finite Elements ; Navier-Stokes Equations ; Numerical Results ; Penalty Model ; Staggered Cylinders ; Engineering ; Engineering General
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: The two-dimensional Navier-Stokes equations and the energy equation governing steady laminar incompressible flow are solved by a penalty finite-element model for flow across finite depth, five-row deep, staggered bundles of cylinders. Pitch to diameter ratios of 1·5 and 2·0 are considered for cylinders in equilateral triangular and square arrangements. Reynolds numbers studied range from 100 to 400, and a Prandtl number of 0·7 is used. Velocity vector fields, streamline patterns, vorticity, pressure and temperature contours, local and average Nusselt numbers, pressure and shear stress distributions around the cylinder walls and drag coefficients are presented. The results obtained agree well with available experimental and numerical data.
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