DIFFUSION POTENTIALS IN MODELS AND IN LIVING CELLS

The behavior of guaiacol resembles that of certain protoplasmic surfaces to such an extent that it can be advantageously used in models designed to imitate certain aspects of protoplasmic behavior. In these models the electrical potentials appear to consist of diffusion potentials and this may be true of certain living cells. In dealing with models we determine ionic mobilities and use these to predict potentials. In studying living cells we measure potentials and from these calculate ionic mobilities. The question arises, how far is this method justified. To test this we have treated guaiacol like a living cell, measuring potentials and from these estimating ionic mobilities. The results Justify the use of this method. This is of interest because the method is most useful in studying protoplasmic activity. In its extended form it enables us to follow changes in mobilities and in partition coefficients due to applied reagents and to metabolism.     

a-c electrical properties of bipolar membranes: Experiments and a model

Summary The a-c electrical properties of bipolar membranes separating equal strength solutions of the same uni-univalent electrolyte are analyzed for the case where both ions have equal mobilities. Two membrane models are treated. In one, the fixed-charge density is assumed to be constant throughout the membrane. In the other, the membrane is regarded as comprising an array of pores separated by walls through which the fixed charge is spread uniformly. Experimental results are reported for the a-c electrical properties of a bipolar membrane prepared from a single polyolephine sheet and immersed in KCl solutions of various concentrations. It is found that the data can be interpreted using the pore model.     

A method to relate steady-state ionic currents, conductances, and membrane potential in ion exchange membranes with unknown thermodynamic properties

A method is presented by which the steady-state properties of an homogeneous, permselective membrane at uniform temperature can be predicted without knowledge of its thermodynamic properties other than assuming that they are functions only of local mole fractions in the membrane. By making this assumption, it is shown how the ionic conductances can be calculated at any point in the membrane from two sets of measurements, (a) R(symm), the steady-state resistance of the membrane measured between identical solutions and (b) V(0), the potential difference between nonidentical solutions for zero current. These two parameters are measured at different external solution compositions (e.g. a varying sodium-potassium ratio ranging from zero to infinity). From these measurements it is shown how the flux equations may be integrated without a knowledge of mobilities, activity coefficients, and other interior membrane parameters. The application of the method to fixed site membranes with variable mobilities is described and the theory for this particular case has also been verified experimentally in glass membranes.1 A possible application to biological membranes is discussed and a comparison is made between the present treatment and previous treatments used to calculate the steady-state properties of cell membranes, notably the theory of Teorell, Meyer, and Sievers and the constant field theory.     

Kinetics of Excitable Membranes : Voltage amplification in a diffusion regime

An understanding of the properties of excitable membranes requires the calculation of ion flow through the membrane, including the effects of nonuniformity in the transverse membrane properties (mobilities, fixed charge, electric field). Permeability is apparently controlled at the external interface. Two factors may be involved here: the statistical blocking of pores by divalent cations, and activation energy. Only the former is included in the present treatment. When the total transmembrane voltage is varied, a redistribution in ionic concentration occurs. This can cause a change in boundary (zeta) potential, large in comparison with the applied voltage change—"voltage amplification." The result is a steep change in membrane conductance. The calculated flow curves are compared with experimental results. The Appendix gives an outline of the numerical method used for solving the boundary value problem with several diffusible ions, across a nonuniform regime.     

APPLICATION OF THE DIFFUSION HYPOTHESIS TO MEMBRANE POTENTIALS

A system consisting of two aqueous solutions, containing equal concentrations of lactic acid, but different concentrations of Na lactate, separated by a layer of amyl alcohol has been described. This system exhibits electrical properties ranging (as the concentration of NaL is increased) from those characteristic of a simple Donnan equilibrium to those characteristic of simple diffusion. The fact that the Donnan P.D. can be treated as a special case of a diffusion potential has been emphasized. The experiments call attention to the effect of the thermodynamic properties of the membrane, and it is suggested that such properties as conductivities, ionic mobilities, and distribution coefficients in membranes of any sort should be investigated. The experiments afford an interesting example of "phase reversal."     

STUDIES ON THE PERMEABILITY OF MEMBRANES : III. ELECTRIC TRANSFER EXPERIMENTS WITH THE DRIED COLLODION MEMBRANE.

The transfer numbers of the ions of electrolytes in the dried collodion membrane, as determined in a previous paper indirectly from the E.M.F. of concentration chains, can also be determined directly by electrical transfer experiments. It is shown that the difficulties involved in such experiments can be overcome. The transfer numbers obtained by the two methods are in satisfactory agreement. The experimental results obtained in the transfer experiments furnish an additional argument in favor of maintaining the theory that the electromotive effects observed in varying concentrations of different electrolytes with the dried collodion membrane may be referred to differences in the mobilities of the anions and cations within the membrane. As was shown by the method of the previous paper, the transfer number depends largely on concentration. There are some minor discrepancies between the values of the transfer numbers obtained by the two methods which, as yet, cannot be completely explained.     

Effect of ionic polarizability on electrodiffusion in lipid bilayer membranes.

Ion-carrier complexes and organic ions of similar size and shape have mobilities in lipid bilayer membranes which span several orders of magnitude. In this communication, an examination is made of the hypothesis that the basis for this unusually wide range of ionic mobilities is the potential energy barrier arising from image forces which selectively act on ions according to their polarizability. Using Poisson's equation to evaluate the electrostatic interaction between an ion and its surroundings, the potential energy barrier to ion transport due to image effects is computed, with the result that the potential energy barrier height depends strongly on ionic polarizability. Theoretical membrane potential energy profile calculations are used in conjunction with Nernst-Planck electrodiffusion equation to analyze the available mobility data for several ion-carrier complexes and lipid-soluble ions in lipid bilayer membranes. The variation among the mobilities of different ions is shown to be in agreement with theoretical predictions based on ionic polarizability and size. Furthermore, the important influence exerted by image forces on ion transport in lipid bilayer membranes compared to the frictional effect of membrane viscosity is established by contrasting available data on the activation energy of ionic conductivity with that for membrane fluidity.     

Regulation of the proton/electron stoichiometry of mitochondrial ubiquinol:cytochrome c reductase by the membrane potential.

The electron transfer reaction catalysed by mitochondrial ubiquinol:cytochrome c reductase is linked to the outwards translocation of protons with an H+ e- stoichiometry of 1 under non-membrane potential condition. The effect of the electrical membrane potential on the H+/e- stoichiometry was investigated. The enzyme was isolated from Neurospora crassa, reconstituted into phospholipid vesicles and electrical membrane potentials of various values were generated across the membranes by means of the valinomycin-induced potassium-diffusion method. Using lithium ions as counterions for the intravesicular potassium, the induced membrane potential was stable for minutes and was not significantly changed by the protons ejected by the working enzyme. This allowed the assay of steady-state reaction rates at pre-given values of electrical membrane potential. The rate ratio between electron transfer and proton translocation declined from 1 to 0.6 with increase of the membrane potential from 0 to 100 mV. The activity of the quinol/cytochrome c redox reaction followed a parabolic dependence, being activated by low (less than 50 mV) potential and inhibited by high (greater than 100 mV) potential. This apparent non-linear dependence was interpreted in terms of a linear flow/force relationship plus a membrane-potential-dependent slip. Evaluation of the parabolic course by means of a modified linear flow/force relation also indicated a decline of the H+/e- stoichiometry from 1 to 0.5 with increase of the membrane potential from 0 to 120 mV. These observations suggest that the membrane potential controls a change of ubiquinol:cytochrome c reductase between two states that have different reaction routes.     

Perturbations in the sof the double layer at an enzymic surface

The surfaces of cells are both charged and enzymically active; furthermore, mass transfer across the surface is occurring constantly. These dynamic processes are capable of perturbing the equilibrium double layer that would be present in the absence of mass transfer and reactions. This paper investigates the influence of enzymic surface reactions on the structure of the diffuse double layer, and conversely the influence of potential on concentration profiles and reaction rates. It is shown that (1) mobility differences in substrate and product can lead to more or less extended double layers and to extrema in the potential profile depending on kinetic factors such as reaction rate and ion mobility of substrate and product and (2) surface reactions can act as a surface concentration switch or amplifier wherein comparatively small variations in bulk concentration produce large variations in surface concentration. Deviations from equilibrium potentials are described by a dimensionless parameter involving reaction rate, ionic strength and the substrate-product mobility difference. Deviations from equilibrium concentrations are described by two electrostatic reaction-diffusion moduli. One of these expresses the effect of differing ion mobilities between substrate and product. Depending on the sign of this parameter, the surface substrate concentration may be either displaced above or below the case (usually hypothetical) of equal ion mobilities. The physiological significance of a reaction or mass flow perturbed surface potential is discussed.     

Are Chironomidae (Diptera) good indicators of water scarcity? Dryland streams as a case study

Water scarcity is becoming one of the greatest challenges that human societies will face during this century. Monitoring water availability is expensive and technically challenging. In this regard, biological communities (e.g. aquatic insects) offer a cost-effective alternative, since they integrate temporal and spatial hydrological variability.Here we explore the potential of Chironomidae (Diptera), which have been usually neglected due to their complex taxonomy, as indicators of both local habitat condition and water scarcity. The study took place in 28 sites across seven dryland streams distributed within a 400 km² section of the Upper San Pedro River basin, southeastern Arizona. The selected streams covered a wide range of hydrological variability, which was continuously measured through the deployment of 15 electrical resistance (ER) sensors. Chironomidae taxa with no drought-resistance strategies were rarely found in streams that experienced frequent drying events (i.e. intermittent and ephemeral streams), suggesting that droughts have the potential to reduce species richness due to local extinctions of drought-intolerant taxa. Intermittent and ephemeral streams registered low canopy cover and a significantly higher abundance of scrapers (which mainly feed on algae) and shredders (feeding on poorly decomposed coarse organic material). This suggests that structural changes associated to drought (e.g. reduced canopy cover and decomposition rates) might lead to changes in the functional composition of the Chironomidae assemblages. We conclude that Chironomidae species can be used as indicators of hydrological variability and the impacts of drought on streams in the absence of flow gauges.     

Increase in lipid microviscosity of unilamellar vesicles upon the creation of transmembrane potential

Summary Diffusion potential of potassium ions was formed in unilamellar vesicles of phosphatidyl choline. The vesicles, which included potassium sulfate buffered with potassium phosphate, were diluted into an analogous salt solution made of sodium sulfate and sodium phosphate. The diffusion potential was created by the addition of the potassium-ionophore, valinomycin. The change in lipid microviscosity, ensuing the formation of membrane potential, was measured by the conventional method of fluorescence depolarization with 1,6-diphenyl-1,3,5-hexatriene as a probe. Lipid microviscosity was found to increase with membrane potential in a nonlinear manner, irrespective of the potential direction. Two tentative interpretations are proposed for this observation. The first assumes that the membrane potential imposes an energy barrier on the lipid flow which can be treated in terms of Boltzmann-distribution. The other interpretation assumes a decrease in lipid-free volume due to the pressure induced by the electrical potential. Since increase in lipid viscosity can reduce lateral and rotational motions, as well as increase exposure of functional membrane proteins, physiological effects induced by transmembrane potential could be associated with such dynamic changes.     

IONIC TRANSFERENCE NUMBERS IN CELLOPHANE MEMBRANES

The "apparent" cation transference number within cellophane is determined for HCl, KCl, NH₄Cl, NaCl, and LiCl. The method consists in measuring the E.M.F. in a concentration chain employing Ag:AgCl electrodes or calomel electrodes and calculating from formulas derived for cases of simple, unconstrained diffusion. The transference numbers and the cation mobilities relative to the chloride ion were found to be higher in the cellophane (relative cation mobilities increased about 40 per cent). The effect of the membrane is discussed. It is emphasized that with the introduction of a membrane as a liquid junction new factors are introduced, which are not considered in the formulas ordinarily used. Such factors may be activity changes due to dimensional or other reasons and particularly electrical effects exhibited by the membrane upon the ionic diffusion. Accordingly the transference number, as determined, may lack well defined physical significance.     

Inferences about membrane properties from electrical noise measurements

Four sources of electrical noise in biological membranes, each with a different physical basis, are discussed; the analysis of each type of noise potentially yields a different sort of information about membrane properties. (a) From the thermal noise spectrum, the passive membrane impedance may be obtained, so that thermal noise measurements are essentially equivalent to the type of since wave analysis carried out by Cole and Curtis. (b) If adequately high frequency measurements could be made, the shot noise spectrum should give information about the average motion of a single ion within the membrane. (c) The number of charge carriers and single ion mobilities within the membrane can possibly be inferred from measurements of noise with a 1/f spectrum. Available data indicate, for example, that increases in axon membrane conductance are not achieved by modulations in the mobility of ions within the membrane. (d) Fluctuations arising from the mechanisms normally responsible for membrane conductance changes can produce a type of electrical noise. Analysis of such conductance fluctuations provides a way to assess the validity of various microscopic models for the behavior of individual channels. Two different probabilistic interpretations of the Hodgkin-Huxley equations are investigated here and shown to yield different predictions about the spectrum of conductance fluctuations; thus, appropriate noise measurements may serve to eliminate certain classes of microscopic models for membrane conductance changes. Further, it is shown how the analysis of conductance fluctuations can, in some circumstances, provide an estimate of the conductance of a single channel.     

Effects of Polarization of the Receptor Membrane and of the First Ranvier Node in a Sense Organ

It has previously been shown that the site of production of the generator potential in Pacinian corpuscles is the receptor membrane of the non-myelinated ending, and the site of initiation of the nerve impulse, the adjacent (first) Ranvier node. Effects of membrane polarization of these sites were studied in the present work. Nerve ending and first Ranvier node were isolated by dissection, electric activity was recorded from, and polarizing currents were passed through them. All observations were done at steady levels of polarization, seconds after onset of current flow. The following results were obtained: The amount of charge transferred through the excited receptor membrane is a function of the electrical gradients across the membrane. The generator potential in response to equal mechanical stimuli increases with resting potential of the receptor membrane. The refractory state of the generator potential is not affected by polarization. The electrical threshold for impulse firing at the first Ranvier node (measured by the minimal amplitude of generator potential which elicits a nodal impulse) is nearly minimal at normal resting potential of the node. Both, hyperpolarization and depolarization lead to a rise in nodal threshold. For any level of polarization of nodal and receptor membrane, the threshold for production of impulses by adequate (mechanical) stimulation appears determined by the generator potential-stimulus strength relation and by the electrical threshold of the node.     

Effect of fire on benthic algal assemblage structure and recolonization in intermittent streams

Abstract: Dry biofilm on rocks and other substrata forms an important drought refuge for benthic algae in intermittent streams following the cessation of flow. This dry biofilm is potentially susceptible to disturbance from bushfires, including direct burning and/or scorching and damage from radiant heat, particularly when streams are dry. Therefore, damage to dry biofilms by fire has the potential to influence algal recolonization and assemblage structure in intermittent streams following commencement of flow. The influence of fire on benthic algal assemblages and recolonization was examined in intermittent streams of the Grampians National Park, Victoria, Australia, using a field survey and manipulative field experiment. The field survey compared assemblages in two intermittent streams within a recently burnt area (within 5 months of the fire) with two intermittent streams within an unburnt area. The two burnt streams were still flowing during the fire so most biofilms were not likely to be directly exposed to flames. Considerable site‐to‐site and stream‐to‐stream variation was detected during the field survey, which may have obscured potential differences attributable to indirect effects of the fire. The manipulative field experiment occurred in two intermittent streams and consisted of five treatments chosen to replicate various characteristics of bushfires that may influence dry biofilms: dry biofilm exposed directly to fire; dry biofilm exposed to radiant heat; dry biofilm exposed to ash; and two procedural controls. After exposure to the different treatments, rocks were replaced in the streams and algae were sampled 7 days after flow commenced. Differences occurred across treatments, but treatment differences were inconsistent across the two streams. For example, direct exposure to fire reduced the abundance of recolonizing algae and altered assemblage structure in both streams, while radiant heat had an effect on assemblage structure in one stream only. The manipulative field experiment is likely to have represented the intensity of a small bushfire only. Nonetheless, significant differences across treatments were detected, so these experimental results suggest that fire can damage dry biofilms, and hence, influence algal recolonization and assemblage structure in intermittent streams.     

Business‐to‐Business Information Technology User Practices at End of Life in the United Kingdom,Germany, and France

Business‐to‐business (B2B) electronics account for a significant volume of the electrical and electronic equipment (EEE) put on the market. Very little B2B waste electrical and electronic equipment (WEEE) is reported as collected in the European Union (EU) in compliance with the WEEE Directive, which uses the policy principle of extended producer responsibility (EPR) to ensure that WEEE is managed correctly. This presents a barrier to parties looking for access to the waste. Company practice dictates the channels into which B2B WEEE flows following primary use. This article presents a study that engaged with company actors directly to get a better understanding of business information technology (IT) EEE asset management. Data were collected to determine the barriers current practice could present to the collection of B2B IT EEE at end of life and the implications of these for the development of policies and strategies for EPR. A questionnaire was developed and data were gathered from organizations in three EU countries—the United Kingdom, Germany, and France—stratified by size. Some notable findings were that there are several routes by which end‐of‐life B2B WEEE can flow. The recycling and refurbishment of B2B IT units at end of use was shown to be commonplace, but it is likely that these units enter streams where they are not reported. The actors disposing of their units did not have information on the management or disposition of these streams. It is concluded that to achieve the goals of EPR for B2B IT WEEE, the networks and the operational practices of these streams need to be better understood when developing strategies and policies.     

Energy flux modulation on the outer membrane of mitochondria by metabolically-derived potential

Voltage-dependent anion channels (VDACs) are the porins in the outer mitochondrial membrane allowing metabolite flux between mitochondria and the cytoplasm. The permeabilities of the VDACs to ATP(-4), ADP(3-), creatine phosphate2-, Pi2-, Pi-, and other charged metabolites depend on the membrane potential. But neither the existence of the electrical potential across the outer membrane of mitochondria, nor its generation mechanisms have been experimentally shown. In this work, the concept of metabolically-derived potential that could be generated on the outer membrane was developed further. The computational study of the quantitative models shows that a steady-state membrane potential above 40 mV may be generated across a membrane with VDACs, if the VDACs are considered to be non-permeable to K+ and Cl-. Free permeability of VDACs to these inorganic ions, mimicking VDACs biological behavior, decreases the potential to nearly 12 mV. This decrease does not result from the electrical shortening of the potential by K+ and Cl- fluxes, but is caused by the electrodynamic compartmentation of the charged metabolites influencing the Goldman fluxes and the enzyme activity determining the fluxes. The interaction of two cyclic steady-state fluxes of charged metabolites due to the synergetic superposition of the potentials generated by each of these fluxes was obtained, and the effect of amplification of one flux by the other was theoretically demonstrated. These calculations based on VDACs' known permeability-voltage characteristics indicate that there is a certain possibility that the cell energy metabolism is regulated on the outer membrane of mitochondria by the electrical potential generated by various metabolically-dependent mechanisms.     

Suppression of flash-induced PSII-dependent electrogenesis caused by proton pumping in chloroplasts

Pre-illumination of the thylakoid membrane of Peperomia metallica chloroplasts leads to a reversible suppression of the flash-induced electrical potential as measured either with the electrochromic bandshift (P515), microelectrode impalement or patch-clamp technique. The energization-dependent potential suppression was not observed in the presence of 1 μ M nigericin suggesting the involvement of proton and/or cation gradients. Energization in the presence of 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) and N,N,N',N'-tetramethylphenylenediamine (TMPD), i.e. cyclic electron flow around photosystem (PS) I, results in the accumulation of TMPD⁺ in the thylakoid lumen. The reversible suppression of the flash-induced membrane potential was not observed in these conditions indicating that it is not a general cation-induced increase of membrane capacitance. Cyclic electron flow around PSI in the presence of DCMU and phenazine methosulfate (PMS) results in the accumulation of PMS⁺ and H⁺ in the thylakoid lumen. The absence of reversible suppression of the flash-induced membrane potential for this condition shows that accumulation of protons does not lead to (1) a reversible increase of membrane capacitance and (2) a reversible suppression of PSI-dependent electrogenesis. Reversible inactivation of PSII by a low pH in the thylakoid lumen is therefore proposed to be the cause for the temporary suppression of the flash-induced electrical potential. The flash-induced PSII-dependent membrane potential, as measured after major oxidation of P700 in far-red background light, was indeed found to be suppressed at low assay pH (pH 5) in isolated spinach ( Spinacia oleracea ) chloroplasts.     

Hysteresis in voltage-gated channels

Ion channels constitute a superfamily of membrane proteins found in all living creatures. Their activity allows fast translocation of ions across the plasma membrane down the ion's transmembrane electrochemical gradient, resulting in a difference in electrical potential across the plasma membrane, known as the membrane potential. A group within this superfamily, namely voltage-gated channels, displays activity that is sensitive to the membrane potential. The activity of voltage-gated channels is controlled by the membrane potential, while the membrane potential is changed by these channels' activity. This interplay produces variations in the membrane potential that have evolved into electrical signals in many organisms. These signals are essential for numerous biological processes, including neuronal activity, insulin release, muscle contraction, fertilization and many others. In recent years, the activity of the voltage-gated channels has been observed not to follow a simple relationship with the membrane potential. Instead, it has been shown that the activity of voltage-gated channel displays hysteresis. In fact, a growing number of evidence have demonstrated that the voltage dependence of channel activity is dynamically modulated by activity itself. In spite of the great impact that this property can have on electrical signaling, hysteresis in voltage-gated channels is often overlooked. Addressing this issue, this review provides examples of voltage-gated ion channels displaying hysteretic behavior. Further, this review will discuss how Dynamic Voltage Dependence in voltage-gated channels can have a physiological role in electrical signaling. Furthermore, this review will elaborate on the current thoughts on the mechanism underlying hysteresis in voltage-gated channels.     

The Non-Steady-State Membrane Potential of Ion Exchangers with Fixed Sites

A system of equations, based upon the assumption that the only force acting on each ionic species is due to the gradient of its electrochemical potential, is used to deduce, in the non-steady state for zero net current, the expression of the difference of electric potential between two solutions separated by an ion exchange membrane with fixed monovalent sites. The membrane is assumed to be solely permeable to cations or anions, depending on whether the charge of the sites is -1 or +1, and not to permit any flow of solvent. Under the assumptions that the difference of standard chemical potentials of any pair of permeant monovalent species and the ratio of their mobilities are constant throughout the membrane, even when the spacing of sites is variable, explicit expressions are derived for the diffusion potential and total membrane potential as functions of time and of solution activities. The expressions are valid for any number of permeant monovalent species having ideal behavior and for two permeant monovalent species having “n-type” non-ideal behavior. The results show that for a step change in solution composition the observable potential across a membrane having fixed, but not necessarily uniformly spaced, sites becomes independent of time once equilibria are established at the boundaries of the membrane and attains its steady-state value even while the ionic concentration profiles and the electric potential profile within the membrane are changing with time.