Translational Diffusion Coefficient and Partition Coefficient of a Spin-Labeled Solute in Lecithin Bilayer Membranes

The translational diffusion coefficient and the partition coefficient of a spin-labeled solute, di-t-butyl nitroxide, in an aqueous suspension of dipalmitoyl lecithin vesicles have been studied by electron spin resonance spectroscopy. When the lecithin is cooled through its phase transition temperature near 41 degrees C, some solute is "frozen out" of the bilayer, and the standard partial molar enthalpy and entropy of partition go more positive by a factor of 8 and 6, respectively. However, the apparent diffusion constant in the lecithin phase is only slightly smaller than that in water, both above and below the transition temperature. The fraction of bilayer volume within which solute is distributed may increase with temperature, contributing to the positive enthalpy of partition. Comparison of time constants suggests that there is a permeability barrier to this solute in the periphery of the bilayer.     

A Simple Method for Measuring of Intestinal Solute Transport in Mucosal Biopsy Specimens

Accurate in vitro measurements of intestinal mucosal solute uptake in humans are often difficult because only small amounts of tissue material are available. We describe a miniaturized everted sleeve method of measuring intestinal solute uptake in endoscopy biopsy samples that combines simplicity, good tissue viability and reproducibility. Biopsies were mounted on a dressmaker needle head stationed immediately over a stirring bar rotating at 1200 rpm. This approach was used to measure taurocholate uptake in sheep and human endoscopy biopsies. Comparison was made to conventional standardized everted sleeve preparations. Na⁺-dependent uptake rates correlated well among individual sheep (R² = 0.88, P < 0.05). There was excellent correlation between conventional and biopsy preparations in humans (R² = 0.98; P < 0.05). The biopsy method overestimated diffusional uptake rates in sheep and humans by two to three fold when compared to conventional everted sleeve preparations. We conclude that this method is valuable to measure Na⁺-dependent solute uptake rates in biopsy samples from human intestine.     

Importance of Body Water in the Efficacy of Convective Solute Transport in Online Hemodiafiltration

High‐volume online hemodiafiltration (OL‐HDF) has been associated with improved patient survival compared to conventional hemodialysis in recent trials, where the importance of convective volume (CV) in this benefit is noted. The purpose of this study was to determine the corporal composition parameters influencing the efficacy of CV in the removal of different molecular weight (MW) molecules. Demographic data, corporal composition parameters with bioimpedance spectroscopy, dialysis features and the reduction rates of different MW molecules in a four‐hour OL‐HDF session were collected in 61 patients. We observed a significant negative correlation of β2‐microglobulin, cystatin‐C, myoglobin and prolactin reduction rates with body surface area, weight, total body extracellular (ECW) and intracellular water (ICW), lean tissue mass and body cellular mass. The multivariable regression analysis identified ECW and ICW as the only corporal composition factors independently associated to the relative elimination of β2‐microglobulin (Beta: –0.801, P = 0.002 for ECW and Beta: –1.710, P = 0.001 for ICW), cystatin‐C (Beta: –0.656, P = 0.010 for ECW and Beta: –1.511, P = 0.004 for ICW) and myoglobin (Beta: –0.745, P = 0.014 for ECW and Beta: –2.103, P = 0.001 for ICW), in addition to CV. Prolactin reduction was only associated with ICW (Beta: –1.540, P = 0.028). When adjusting CV with ECW and ICW, only the ratio CV/ECW was an independent predictor for higher elimination of β2‐microglobulin, cystatin‐C and myoglobin. The corporal composition parameters independently associated to the reduction of medium‐sized molecules are the extracellular and intracellular water. The ratio “convective volume/extracellular water” predicts higher efficacy of convective transport. Adjusting the convective volume to patient features could be useful to monitor the efficacy of OL‐HDF and to prescribe individualized therapies.     

Cooperative Effects in Models of Steady-State Transport across Membranes, I

Three simple models are discussed which show cooperative effects in the steady-state transport of neutral molecules across a membrane. The Bragg-Williams approximation is employed, and its limitations discussed. In this approximation to cooperative problems, the diagram method for the calculation of steady-state probabilities and fluxes can still be used. This paper is closely related to one by Blumenthal, Changeux, and Lefever, and provides the foundation for sequels on an oscillatory steady-state phase-transition model (part II) and on simple models possibly related to steady-state potassium ion transport across a nerve membrane (part III).     

On the Theory of Ion Transport Across the Nerve Membrane, VII. Cooperativity Between Channels of a Large Square Lattice

The exact kinetics of a 10 x 10 Ising system (periodic boundary conditions) with two-state channels arranged in a square lattice was studied by computer simulation. With all three values of the cooperativity parameter used, no induction in the K(+)-current curve was obtained. This confirms one of our previous conclusions concerning K(+) channels in the squid axon membrane: models with interacting channels arranged in a twodimensional lattice (Adam's model) are apparently excluded. Other topics included: equilibrium properties; short-range pair correlation functions; phase transition.     

Influence of Spin Coating Parameters on Gas Transport Properties of Thin-Film Composite Membranes

The influence of casting centrifugation process parameters, such as a rotation speed (ω), the amount of the film-forming solution (V), and its concentration (C) on transport properties of composite membranes were investigated. A number of composite membranes based on poly (1-trimethylsilylpropyne) (PTMSP) and micro- (MFFK-1) and ultrafiltration (UFFK) membranes were obtained using the spin-coating method. For the first time, an unexpected dependence of permeance and ideal selectivity on rotation speed had been discovered: the thickness of the selective layer decreases from 3.0 to 1.0 μm for MFFK-1 and from 1.7 to 1.1 μm for UFFK with an increase of spin coater rotation speed from 500 to 3000 rpm. However, the gas permeance of composite membranes in the range of 500–2000 rpm was reduced due to an increase of a penetration depth of PTMSP into a support layer porous structure (estimated by the EDX method). The permeance of the PTMSP/UFFK membranes was higher than PTMSP/MFFK-1 membranes due to a thinner selective layer and a lower penetration depth of polymer solution into the pores of the support. The highest CO₂/N₂ selectivity values were achieved as 5.65 ± 0.9 at CO₂ permeance 5600 ± 1000 GPU for PTMSP/UFFK membranes (C_PTMSP = 0.35%, V_solution = 1 mL, ω = 1000 rpm), and 6.1 ± 0.5 at CO₂ permeance 4090 ± 500 GPU for PTMSP/MFFK-1 membranes (C_PTMSP = 0.35%, V_solution = 1 mL, ω = 2000 rpm).     

Cooperative Effects in Models of Steady-State Transport across Membranes, III. Simulation of Potassium Ion Transport in Nerve

An extremely simple membrane transport model simulates qualitatively the observed flux-potential curves for steady-state potassium ion transport across a nerve (squid) membrane in the special case of high external potassium ion concentration. The transporting units (protein molecules) in the model are assumed to exist in two different conformations, which interact cooperatively. The membrane potential induces a transition between the two conformations by virtue, primarily, of a polarizability difference in the two conformations. Differential proton binding may make an important contribution to this effect.     

Activities of Transport Adenosine Triphosphatases in Erythrocyte Membranes of Healthy and Hypertensive Subjects

The Na⁺, K⁺-ATPase activity in the erythrocyte membrane was measured in 25 untreated essential hypertensive patients and 25 age-matched healthy normotensive subjects. In addition, the Ca²⁺, Mg²⁺-ATPase activity was measured in 20 hypertensive and 25 age-matched normotensive subjects. The Na⁺, K⁺-ATPase activity of healthy Chinese measured in this study was similar to the data reported in a Dutch study. We therefore could not support a theory which speculated an ethnic influence on Na⁺, K⁺-ATPase activity. Both Na⁺, K⁺-ATPase and Ca²⁺, Mg²⁺-ATPase activities were slightly lower in hypertensive patients as compared with those in normotensive subjects, but the differences were not statistically significant. We concluded that the transport ATPase activities were not impaired in the erythrocyte membranes of hypertensive patients.     

Demonstration of Electrogenic Na+-dependent D-glucose Transport in Intestinal Brush Border Membranes

Na(+)-coupled D-glucose transport was studied in isolated membrane vesicles from intestinal brush borders. Concentration gradients of SCN(-), K(+), and H(+) were established between the intravesicular solution and the incubation medium and their influence on D-glucose uptake from the medium was measured. A gradient (medium > vesicle) of NaSCN, but not of KSCN, produced a transient overshoot of D-glucose uptake above the equilibrium level. Similarly, an increase of the membrane conductance with valinomycin (K(+)-conductance) or with uncoupling agents of oxidative phosphorylation (H(+)-conductance) induced an overshooting D-glucose uptake, provided a (vesicle > medium) K(+)-gradient or a H(+)-gradient, respectively, was present in each case. The transient overshoot is evidence that D-glucose was taken up against its concentration gradient (up to 10-fold). The gradients of SCN(-), K(+) (in the presence of valinomycin), and H(+) (in the presence of uncouplers) are thought to contribute to the "driving" force for this "active" D-glucose transport by changing the electrical potential across the vesicle membrane and thus making the inside more negative (with respect to the medium). These experiments, therefore, provide evidence that the Na(+)-coupled D-glucose translocation across the brush border membrane is an electrogenic process, i.e., the positive charge associated with Na(+) is not compensated by the co-movement of an anion or the counter-movement of a cation via the glucose "carrier". The results imply that an electrical potential across the brush border membrane may play an important role in determining the transport of D-glucose by intact cells.     

Sustained Administration of Hormones Exploiting Nanoconfined Diffusion through Nanochannel Membranes

Implantable devices may provide a superior means for hormone delivery through maintaining serum levels within target therapeutic windows. Zero-order administration has been shown to reach an equilibrium with metabolic clearance, resulting in a constant serum concentration and bioavailability of released hormones. By exploiting surface-to-molecule interaction within nanochannel membranes, it is possible to achieve a long-term, constant diffusive release of agents from implantable reservoirs. In this study, we sought to demonstrate the controlled release of model hormones from a novel nanochannel system. We investigated the delivery of hormones through our nanochannel membrane over a period of 40 days. Levothyroxine, osteocalcin and testosterone were selected as representative hormones based on their different molecular properties and structures. The release mechanisms and transport behaviors of these hormones within 3, 5 and 40 nm channels were characterized. Results further supported the suitability of the nanochannels for sustained administration from implantable platforms.     

The Role of Cytoplasmic Membranes in Controlling the Transport of Nuclear Messenger RNA and Initiation of Protein Synthesis

Three different classes of poly(A)-containing RNA have been identified in the yeast system. They have been characterized by their kinetics of labeling, by their localization in the nuclear, membranous, and soluble cytoplasmic fractions, and by their size. The evidence indicates that the membrane-bound poly(A)-containing RNA is a product of the nuclear poly(A)-containing RNA and is the precursor of the polysomal poly(A)-containing RNA. In addition, it has been demonstrated that transport of mRNA is coupled with protein synthesis.     

Studies on Calcium Transport and Calcium-dependent Adenosine Triphosphatase Activity of Erythrocyte Membranes in Hereditary Spherocytosis

Evidence has been recently presented of a relative deficiency of Ca2+ - dependent adenosine triphosphatase activity of erythrocyte membranes obtained from patients with hereditary spherocytosis. We have sought to confirm these findings by measuring calcium efflux from intact erythrocytes of patients with hereditary spherocytosis, as well as erythrocyte calcium concentrations, but find both these parameters to be normal. Ca2+-dependent adenosine triphosphatase activity, as well as Ca2+ -dependent membrane phosphorylation was also not found to be deficient in erythrocyte membranes from subjects with hereditary spherocytosis. These studies do not support the postulate that an accumulation of calcium affects the deformability of erythrocytes and their subsequent destruction in the spleen.     

Analysis of a Model for Active Transport

A particular model of active transport is analyzed, at steady state, by means of a diagram method for the kinetics, and supplemented by use of a free energy diagram. It is assumed that there is cyclic adsorption of adenosine triphosphate (ATP), chemical reaction to adenosine diphosphate (ADP), and desorption of ADP, on a two-state protein (the "carrier"). ATP is the "effector" for the change in state of the protein.     

On the Theory of Ion Transport Across the Nerve Membrane, VI. Free Energy and Activation Free Energies of Conformational Change

Empirical functions, such as n(infinity)(V) and tau(n)(V) (of the Hodgkin-Huxley type), can be recast in terms of more fundamental functions F(V) (related to a conformational free energy change) and theta(V) (related to the corresponding free energies of activation). Examples of F(V) and theta(V) are given, for squid and frog node. F(V) is essentially a quadratic function of V. The possible molecular origin, for protein-like subunits, of the linear (e.g., net charge) and quadratic (e.g., polarizability) terms in F(V) is discussed. The F(V), theta(V) kind of analysis leads rather automatically to a simple explanation of the well-known approximate coincidence in location (V value) of the maximum in tau(n)(V) (time constant) and the steeply rising part of n(infinity)(V) (also m, 1 - h).