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This paper presents the results of constant-current (chronopotentiometric) measurements of the egg yolk phosphatidylcholine (PC) bilayer membrane without and with cholesterol. The experiments were performed on planar bilayer lipid membrane (BLM) formed by the Mueller-Rudin method. It is demonstrated that the constant-intensity current flow through bilayer membranes generated fluctuating pores in their structure. The presence of cholesterol in the membrane caused an increase in the value of the breakdown potential. It is postulated that greater stability of the bilayer with cholesterol can result from an increased critical pore radius (at which the bilayer would undergo irreversible rupture). This confirms that cholesterol has a stabilizing effect on BLM. Besides, our results suggest that addition of cholesterol causes shift in the distribution of pore conductance towards a smaller value. It is suggested that this can be connected with the phenomenon of domain formation in the membranes containing high concentration of cholesterol. Moreover, it is shown that chronopotentiometry with programmable current intensity is a promising method for observation of the membrane recovery process.

Stochastic properties of a fluctuating nanopore generated and sustained by an electric field in a lipid bilayer membrane are studied. It is shown that the process of voltage fluctuations, in the current clamp experiment, is a stochastic fractal with long memory, which is the main reason for its nonstationarity. The aging process contributes to the nonstationarity if molecular interactions in the membrane are weak. An attempt to classify the process reveals a non-Gaussian distribution with long tails, which contradicts the hypothesis of fractional Brownian motion, showing that stable motion may be possible. The self-similarity index, estimated by three different methods, depends on current value and membrane sensitivity to electric field in a well defined and explicable manner. The stochastic analysis provided for calculated conductance of nanopore revealed the process close to 1/f noise, the result observed only for the pores not exceeding 1 nm in diameter, induced in membranes with strong molecular interactions. Our results show that such a pore is the simplest biological system needed for flicker noise to occur, and the complexity of highly regulated protein channel is not a necessary factor. A case of noise 1/f(2), observed for a pore with impeded dynamics, suggests a process without memory in such a situation. A physical interpretation is presented for some of the results.

This paper presents the application of chronopotentiometry in the study of membrane electroporation. Chronopotentiometry with a programmable current intensity was used. The experiments were performed on planar bilayer phosphatidylcholine and cholesterol membranes formed by the Mueller-Rudin method. It was demonstrated that a constant-intensity current flow through the bilayer membranes generated voltage fluctuations during electroporation. These fluctuations (following an increase and decrease in membrane conductance) were interpreted as a result of the opening and closing of pores in membrane structures. The decrease in membrane potential to zero did not cause the pore to close immediately. The pore was maintained for about 200 s. The closing of the pore and recovery of the continuous structure of the membrane proceeded not only when the membrane potential equalled zero, but also at membrane potentials up to several tens of millivolts. The fluctuations of the pore were possible at values of membrane potential in the order of at least 100 mV. The size of the pore changed slightly and it closed after some time below this potential value.

The effects of alpha-tocopherol on electric properties of bilayer lipid membranes were investigated. Planar bilayer membranes formed by the Mueller-Rudin method were used. Voltammetric and chronopotentiometric measurements were performed using a four-electrode potentiostat-galvanostat. It was demonstrated that registration of membrane capacitance, resistance, and voltammetric characteristics provided information about the change in the structure and permeability of bilayer lipid membranes. The results suggested that incorporation of alpha-tocopherol into lipid membrane destabilized its structure and facilitated the electrogeneration of pores. The possible role of observed changes in physiological functions of alpha-tocopherol was discussed.

The constant-intensity current chronopotentiometric measurements of egg yolk phosphatidylcholine bilayer membranes (BLM) are presented. It is demonstrated that a constant-intensity current flowing through the bilayer membranes generates the pores in their structures. For the current intensity from 0.1 to 2.0 nA, the generated pores open and close cyclically. The frequency of oscillations depends on the current intensity: the higher current intensity, the higher frequency of pore oscillations. It is suggested that the presented method may allow to create one pore in BLM and to observe its dynamical behaviour. Based on chronopotentiometric curves, a method of pore conductance calculations is presented. It is demonstrated that the value of obtained conductance can be applied for pore diameter estimation. The hypothetical application of constant-current method as a biotechnological tool for selective and controlled incorporation of molecules into microorganisms is discussed., (Copyright 1998 Elsevier Science B.V.)

A new method and a new apparatus for capacitance measurements on bilayer lipid membranes are described. The membrane is charged and discharged with a constant current during the measurement. The charge-discharge cycle duration, which is proportional to the membrane capacitance, is measured. The measured time period is converted into a binary number by digital systems and then this number is either further converted into a constant capacity-proportional voltage or read out by the computer. The apparatus makes it possible to measure the capacitances of voltage-polarized membranes. Application of the apparatus to capacitance measurements of bilayer lipid membranes during their potential on the capacitance is presented. The capacitances of membranes stimulated by rectangular voltage pulses and of those stimulated by a linearly varying potential were reported.