Different effects of two Poloxamers (L61 and F68) on the conductance of bilayer lipid membranes.

The integral conductance of planar lipid bilayer membranes in the presence of two Poloxamers (Pluronics) L61 and F68 with the same lengths of hydrophobic poly(propylene oxide) blocks and the different lengths of hydrophilic poly(ethylene oxide) blocks increases with an increase in the concentration of both Pluronics; however, the shape of the conductance-concentration curves is super linear for L61 and sublinear for F68. In the presence of both Pluronics, rare discrete current jumps are observed against the background of continuous current. At high concentrations, the I-V curves of membranes with both L61 and F68 became nonlinear at sufficiently low voltages but differed significantly. At voltages greater than 50 mV, the conductance of membranes with L61 increased sharply and quantized jumps were observed toward higher conductance, which could be interpreted as the appearance of additional pores. On the contrary, the conductance of membranes with F68 decreased and quantized jumps to lower conductance were observed, which could be interpreted as blocking of already existing pores. We attributed the differences in the conductance-concentration and I-V curves of these two Pluronics to their different effects on the dynamics of membrane hydration and, accordingly, on the probability of formation of conducting pores.

Lipoic acid nanoforms based on phosphatidylcholine: production and characteristics.

We describe the dynamics of lipoic acid (LA) alone, incorporated in liposomes and as a part of nanoemulsions. Mass spectrometry shows that LA in water forms aggregates of two or three molecules in the form of a negatively charged ion and a neutral molecule. Phosphatidylcholine (PC)-based nanoforms of LA as liposomes and nanoemulsions with a particle size equal to 145 nm are characterized by a high degree of incorporation of LA into the nanoparticles and long-term stability during storage at room temperature. Dynamic light scattering (DLS) gives the polydispersity index of the nanoforms (> 0.3), characterizing the homogeneity of the obtained nanodispersions. We found that such emulsions can significantly (5 ×) increase the concentration of LA in the aqueous phase (5-7 mg/mL) when compared with an aqueous solution of LA (1 mg/mL) and by 40% when compared with PC liposomes (4 mg/mL). Moreover, the inclusion of LA in liposomes and nanoemulsions from PC did not change the neutral ζ-potential characteristic of PC nanoforms. CryoTEM established that the structural organization of the liposomes practically did not differ from nanoemulsions and both nanoforms contained both multilayer and single-layer vesicles. When studying the release kinetics of LA from phosphatidylcholine nanoforms, we found that at 22 h, 45-55% of LA was released from nanoparticles, but that at the initial stage of the process LA was slowly released from the nanoemulsions and rapidly from the liposomes. Conductance measurements indicate that LA delivered in all the three forms increase membrane permeability, though this result is most marked with the LA in PC liposomes.

Estimation of the parameters of the Smoluchowski equation describing the occurrence of pores in a bilayer lipid membrane under soft poration.

The conductive lipid pores occurring in planar bilayer membranes are known to manifest themselves experimentally as current fluctuations. Reliable recording of such fluctuations during phase transitions, as well as in membranes with various additives (for example, SDS), allows one to determine the characteristics of hypothetical hydrophilic pores, namely, their number, sizes, lifetimes, and duration of time intervals between pores. Because, in contrast with electroporation, the emergence of pores in a membrane does not require high voltages, this process is called soft poration. Studying the characteristics of pores under soft poration allows us to estimate the parameters of the Smoluchowski equation and compare them with the corresponding parameters used to describe electroporation. In this work, the experimental characteristics of current fluctuations in the membrane with the addition of SDS to the bulk solution were used to estimate the parameters of the Smoluchowski equation: the pore edge tension, the energy of the hydrophobic pore/hydrophilic pore barrier, the coefficient of pore diffusion in the radius space, the initial distribution density of the number of pores, and the attempt rate density of the lipids in a membrane. The obtained estimates are close to the parameter values used in studies of electroporation.

Erlang flow of hydrophilic pore formation and closure events in a lipid bilayer during phase transition resulting from diffusion in the radius space.

The Smoluchowski equation with an energy profile of a special type and an assumed hydrophobic ("half") pore source term is used to describe the process of hydrophilic pore formation in a lipid bilayer at the gel-liquid phase transition. The source term reflects the occurrence of molecule packing defects in a lipid bilayer at phase transition. The time sequences of the pore formation and closure events are treated as non-stationary, second-order Erlang flows whose characteristics depend on the equation solution. The computed distributions of the time intervals between hydrophilic pores, and pore lifetimes agree with the previously published experimental interpulse interval and pulse duration histograms for the current fluctuations through planar bilayer membranes of DPPC immersed in a LiCl aqueous solution containing polyethylene glycol. Thus, the statistical analysis of pore formation and closure times leads us to conclude that firstly, the increased permeability of a lipid bilayer during the gel-liquid phase transition is accounted for by the emergence of additional hydrophobic defects in the heterogeneous structure of the bilayer and secondly, that the non-exponential distributions of the lipid channel closed and open times observed in experiments are evidence that the process of hydrophilic pore formation is not a one-step process but involves at least two dependent events.

[Lipid pore states in bilayer lipid membranes at phase transition in LiCl solution with addition of polyethylene glycols].

Statistics on open and closed states of the lipid pores was experimentally investigated in planar lipid bilayer membranes from dipalmitoylphosphatidylcholine at the phase transition temperature in LiCl solution (1 M). Polyethylene glycol of different molecular mass was added to the solution. Transmembrane current pulses were measured at voltage-clamp conditions (50 mV). The histograms of the pore lifetime and the pulse interval have no exponential shape but the pronounced maximum. It means that the pulse sequence of the transmembrane current measured in the phase conditions is dependent. The emergence of the new pulses depends on the emergence time of the previous pulses. The time of the pulse end depends on the time of the pulse start. The proposed Erlang model approximates the experimental histograms quite accurately. According to the model the lipid pore can have several open and closed states.

[Acoustic thermometry of the patient brain with traumatic brain injury].

Non-invasive deep brain acoustic thermometry is carried out for two patients at Burdenko Neurosurgery Institute. This method is based on the measurements of the own thermal acoustic radiation of the investigated object. These two patients have got the brain injury. Some of their skull bones are absent. Infrared thermometry was also used to measure the surface temperature of the forehead skin. On the basis of the experimental data the temperatures deep within the brain were reconstructed. The values for the two patients are equal to 37.3 0.7 and 37.0 0.3 degrees C.

Effect of colloidal magnetite (Fe3O4) nanoparticles on the electrical characteristics of the azolectin bilayer in a static inhomogeneous magnetic field.

This work is devoted to the study of the combined effects of applied magnetic field and MNPs on the electrical characteristics of bilayer lipid membranes. We present results of the study of electrical parameters of azolectin membranes in a static inhomogeneous magnetic field at the one-sided addition of positively charged quasi-spherical superparamagnetic magnetite nanoparticles with a diameter of about 4 nm. The magnet was located at different distances from the membrane, and the magnetic field attracted the nanoparticles to the membrane surface with different strengths. We observed three pronounced effects that depended on the external magnetic field. Firstly, after addition of nanoparticles in a magnetic field, the conductance of the membranes increased. A smooth increase in conductance was accompanied in some cases by the appearance of current jumps, which can be associated with the formation of through pores with a radius of no more than 1 nm. The conductance increased with increasing magnetic field gradient. Secondly, at zero command voltage, a negative current through the membrane was observed. Although our experiments did not allow us to unambiguously determine which particles create this current, we believe that this current is associated with the penetration of particles through the membrane. This effect intensified with increasing magnetic field gradient. Thirdly, we observed a sharp change in the nonlinear dependence of capacitance on voltage associated both with the change in the surface potential of the azolectin membrane and with the effect of MNP binding to the membrane surface on the apparent membrane capacitance.

Passive Acoustic Thermometry of the Chest of a Person with COVID-19.

Passive acoustic thermometry (PAT) was used to study the dynamics of changes in the chest temperature of a person with COVID-19 over the course of about two and a half weeks after quarantine. PAT, which can measure deep body temperature, showed that the integral temperature of tissues surrounding the lungs increased from 32.2 ± 0.07 to 33.0 ± 0.03°C about 10 days after the end of quarantine. This may indicate increased blood supply to the lungs, i.e., an indication of recovery. Infrared thermometry used to monitor recovery yielded no results.

Increase in the current variance in bilayer lipid membranes near phase transition as a result of the occurrence of hydrophobic defects.

Most researchers associate the increase in the permeability of lipid bilayers of artificial and biological membranes observed in various experiments with the formation of hypothetical hydrophobic and hydrophilic pores. Although the existence of hydrophobic defects, as the first stage of the formation of a hydrophilic pore, was hypothesized decades ago from electroporation experiments, the difficulty of describing this stage is determined by the lack of experimental data confirming the existence or at least associated with hydrophobic pores. We explored the increase in the current variance through the lipid membrane, observed when approaching the phase transition from the side of high temperatures, and have associated it with capacitive currents arising in response to the formation of hydrophobic pores. Assuming that the number of hydrophobic pores in a membrane follows a Poisson distribution, and thus, the mean number of hydrophobic pores is equal to the variance of that number, we used the measurements of the membrane current variance to evaluate the number of hydrophobic pores. Analysis of experimental data within this model allows us to estimate the number of hydrophobic pores at or above the phase transition and shows that the number of hydrophobic pores in a membrane close to the phase transition increased 20 times compared to the number of hydrophobic pores existing in the membrane far from the melting transition.

The effects of SDS at subsolubilizing concentrations on the planar lipid bilayer permeability: Two kinds of current fluctuations.

Detergent effects on lipid bilayers of artificial and biological membranes at subsolubilizing concentrations are known to include the membrane permeabilization which manifests itself through both a flip-flop of detergent molecules from the outer monolayer to the inner monolayer and the membrane leakage of entrapped solutes. We have explored the current fluctuations occurring in planar BLM of asolectin in the presence of ionic detergent SDS at subsolubilizing concentration. Two groups of current fluctuations which the average duration differs by two orders of magnitude can be distinguished. We assume that these differences in the duration of current fluctuations are associated with a different number of SDS molecules in the walls of the putative toroidal hydrophilic pores. We associated short pulses with the formation of short-lived lipid hydrophilic pores. Impulses of greater duration (steps) were associated with the formation of hydrophilic pores, the walls of which contain detergent. Taking into account the characteristics of these pores, we estimated the pore energy, as well as the edge energy of these two kinds of pores. We believe that the flip-flop of SDS molecules in liposomes is provided by long-lived pores, and the contents of the liposome leakage occurs through all pores.

["Blocking" by polyethyleneglycol of single lipid pores arising in unmodified bilayer lipid membranes during phase transitions].

Changes in the ionic permeability of bilayer lipid membranes from dipalmitoyl phosphatidylcholine at temperatures of phase transition in LiCl (1 M) solution after the addition of polyethyleneglycols of different molecular masses have been studied. The transition of ionic membrane channels from the conducting state to a blocking nonconducting state using polymers makes it possible to calibrate lipid pores. It was shown that low-molecular-weight glycerol, polyethyleneglycols with molecular masses of 300 and 600 decrease the amplitude of fluctuations of the current through the membrane, the decrease being proportional to the size of the polymer molecule being incorporated. The addition of polyethyleneglycols with molecular masses of 1450, 2000, and 3350 decrease the current fluctuations to the basal noise level. This result is considered as a complete blockade of ion channel conductivity. In the presence of rather large polymers, such as polyethyleneglycols with molecular masses of 6000 and 20000, which are practically not incorporated into the pore, single current fluctuations occur again; however, their amplitudes are somewhat smaller than in the absence of polyethyleneglycol. It is assumed that the complete blockade of the conductivity of lipid ionic channels by polyethyleneglycols with molecular masses of 1450, 2000, and 3350 is due to the dehydration of the pore gap and the conversion of the hydrophilic pore to a hydrophobic pore.

Physical and computer-based modeling in internal temperature reconstruction by the method of passive acoustic thermometry.

The purpose of this work was to investigate experimentally the capacity of passive acoustic thermometry (PAT) for the reconstruction of 1D, time-variable distributions of the internal temperature. Because in the PAT a noise signal is measured, a considerable integration time (about one minute) is required to attain an acceptable error level (0.5-1K). To optimize the time, an algorithm was proposed to take account of the fact that the temperature satisfied the heat equation. The problem was reduced to that of determining two parameters (initial temperature and thermal diffusivity) of the object under study. The desired parameters were considered constant and were not determined anew after each measurement; instead, their values were refined using all the previous measurements. The proposed algorithm was tested experimentally (where the temperature was reconstructed in a model object, a slab of polytetrafluoroethylene) and investigated by means of computer modeling. The duration of one measurement was about 5.5s. As a result, an error of the temperature reconstruction of about 0.5K, acceptable for medical applications, was attained after 30-60s (depending on the depth) from the beginning of the measurements. After that, temperature distributions can be reconstructed after each measurement without loss of the reconstruction accuracy. The proposed method can be used to control the temperature under a local hyperthermia, lasting 1 min and more, of the human body.

[On measurements of higher harmonics of transmembrane current].

Measurements of higher harmonics of transmembrane current in bilayer lipid membranes from diphytanoyl phosphatidylcholine (DPhPC) in n-decane and n-tetradecane, caused by alternating voltage applied to the membrane, have been conducted. A universal relation between the amplitudes of harmonics was suggested and experimentally checked. This allowed one to calculate the coefficients of expansion of membrane capacity in series with even powers of membrane voltage and to compare the inhomogeneity of membranes from diphytanoyl phosphatidylcholine in n-decane and n-tetradecane with respect to thickness.

[A study of the inhomogeneity of bilayer lipid membranes by measurements of higher harmonics of transmembrane current].

Higher harmonics of alternating current in bilayer lipid membranes caused by sinusoidal voltage applied to the membrane were measured. The bilayer lipid membranes were prepared from diphytanoylphosphatidylcholine in n-decane and n-tetradecane, and measurements were conducted with the aid of an analog-to-digital converter of 16th category. Sinusoidal voltage was formed using a digital-to-analog converter of the 16th category. The dynamic region of measurements was up 90 dB. The results of measurements were used to determine the alpha and beta coefficients of the expansion of membrane capacity C in terms of membrane voltage U C = C0 (1 + alphaU2 + betaU4). We showed in the framework of the electrostriction model that the relation between the alpha and beta coefficients characterizes the inhomogeneity of bilayer lipid membrane with respect to its thickness and Young modulus of elasticity.

[A soft poration of planar bilayer lipid membranes from dipalmitoylphosphatidylcholine at the temperature of the phase transition from the liquid crystalline to the gel state].

A method of soft poration of lipid bilayer was suggested, which is based on the structural rearrangement of lipid bilayer formed from disaturated phospholipids on the phase transition from liquid crystalline state to the gel. As opposed to the widely used method of electropbration, this method allows one to obtain a lipid pore population without application of high electric field. In the case of soft poration, the electric field does not exceed the physiological level of 10-100 mV. It was shown that, in planar bilayer lipid membranes formed from dipalmitoylphosphatidylcholine in water solution of 1 M LiCl, there appear up to 10 lipid pores in 1 min per 1 mm of membrane surface with an average conductivity of a pore of 31 +/- 13 nS. The average pore radius estimated using soluble polyethylene glycols ranged between 1.05-1.63 nm. Monovalent cation conductivity of a single lipid pore on soft poration was shown to decrease in the order Li+ > or = Na+ > K+ = Rb+ > or = Cs+. This order coincides with that observed by Marra and Israilashvili for dipalmitoylphosphatidylcholine-water interbilayer where the repulsive hydration force contribution is significant.

Fundamentals and prospects of passive thermoacoustic tomography.

The problems and possibilities of passive thermoacoustical tomography are discussed. Algorithms for reconstruction of internal temperature in the human body are proposed. These algorithms take into account heat transfer and blood circulation and the absorption factor, obtained previously. The results of reconstruction of deep temperature in the human hand in simulations with the medium with a heated object are reported. These results support the possibility of the correlation measurements of the thermal acoustic radiation. Such measurements allow the information on ultrasound absorption by the object under study to be obtained and open the way to the development of a passive acoustic tomography system using a priori information on the absorption factor.

[Kinetics of resealing of lipid pores]. / Kinetika zatekaniia lipidnykh por.

The resealing of lipid pores in BLM at the temperature of phase transition was studied. It is show that the size of a pore correlates with the time of its resealing. The correlated coefficient of pore diffusion is close to that of lipid molecules in the gel state.

[Exponential distribution of interimpulse current intervals in lipid bilayer membrane at phase transition temperature]. / Eksponentsial'noe raspredelenie mezhimpul'snykh intervalov toka v bisloinykh lipidnykh membranakh pri temperature fazovogo perekhoda.

The statistical analysis of current fluctuations in unmodified bilayer lipid membranes at the phase transition temperature was made. An exponential distribution of current fluctuations was revealed.

[Electric capacitance of bilayer lipid membranes from hydrogenated egg lecithin during phase transition from liquid crystalline state to gel]. / Elektricheskaia emkost' bisloinykh lipidnykh membran iz gidrirovannogo iaichnogo letsitina pri fazovom perekhode iz zhidkokristallicheskogo sostoianiia v gel'.

The electrical capacity of planar bilayer lipid membranes (BLM) from natural hydrogenated egg lecithin (HEL) in n-decane at a temperature of phase transition was measured. The temperature of phase transition was determined calorimetrically to be 51 degrees C. The data obtained revealed a phase separation of HEL in BLM into two fractions, one freezing at 42-44 degrees C and one that is converted to a liquid-crystal state at 51-59 degrees C. It was assumed that the first fraction is rich in dipalmitoyl lecithin, and the second fraction is rich in distearoyl lecithin. Freezing and the transition to the liquid-crystal state were accompanied by an increase and decrease in membrane thickness, respectively, in part due to a displacement of the solvent from the torus to the planar part of the bilayer. The displacement of the solvent is explained by changes in the disjoining pressure in BLM, which arises across the lipid bilayer due to van der Waals forces of attraction between water layers on both sides of the BLM.