Ryszard Olchawa

Membrane fluidity and the surface properties of the lipid bilayer: ESR experiment and computer simulation

Penetration of the liposome membranes formed in the gel phase from DPPC (DPPC liposomes) and in the liquid-crystalline phase from egg yolk lecithin (EYL liposomes) by the TEMPO (2,2,6,6-tetramethylpiperidine-1-oxyl) and 16 DOXYL (2-ethyl-2-(15-methoxy-oxopentadecyl)-4,4-dimethyl-3-oxazolidinyloxy) spin probes has been investigated. The penetration process was followed by 120 hours at 240C, using the electron spin resonance (ESR) method. The investigation of the kinetics of the TEMPO probe building into the membranes of both types of liposomes revealed differences appearing 30 minutes after the start of the experiment. The number of TEMPO particles built into the EYL liposome membranes began to clearly rise, aiming asymptotically to a constant value after about 100 minutes, whereas the number of the TEMPO particles built into the DPPC liposome membranes was almost constant in time. The interpretation of the obtained experimental results was enriched with those of computer simulation, following the behavior of the polar heads (dipoles) of the lipid particles forming a lipid layer due to the change in the value of the model parameter, k, determining the mobility of the dipoles. The possibility of the formation of an irregular ordering of the polar part of lipid membranes was proved, which leads to the appearance of spaces filled with of water for k > 0.4. The appearance of these defects enables the penetration of the bilayer by the TEMPO particles. The limited mobility of lipid polar heads (k < 0.2) prevents the appearance of such areas facilitating the penetration of the lipid membrane by alien particles in the gel phase.

Two-step impact of Amphotericin B (AmB) on lipid membranes: ESR experiment and computer simulations

Abstract In this study, the electron spin resonance (ESR) method was used to examine the effect of Amphotericin B (AmB) molecules on the fluidity of model membranes made of dipalmitoylphosphatidylcholine (DPPC). The changes occurring under increased AmB concentrations in the spectroscopic parameters of spin probes placed in liposomes were determined. Three probes were used, penetrating the membrane at different depths which allowed the changes in its fluidity to be found in the transverse section. A computer model of the surface layer of membrane, with AmB admixture, was developed and subjected to computer simulation. The effect of changing concentration of the admixture on the binding energy in the system of dipoles representing the surface of the membrane was examined. The ESR studies showed that the process of accumulation of AmB molecules in the membrane has two stages, marked by local maxima in the ESR spectra. The first appears for concentrations of ca. 0.25–0.5% and the second appears for ca. 2.5–3% AmB of its molar ratio to DPPC. The computer simulations permitted reconstructing the two-stage mechanism of interaction between the molecules and the membrane. They demonstrated that, at low concentrations, the AmB molecules position themselves flat on the membrane surface. After the threshold concentration is exceeded, they re-orientate to a vertical position. This process leads to the perforation of the membrane.

The impact of humic and fulvic acids on the dynamic properties of liposome membranes: the ESR method

Abstract This paper presents the results of research on the influence of two fractions of humic substances (HS): fulvic acids (FA) and humic acids (HA), as a function of concentration, on the liposome membranes formed from egg yolk lecithin (EYL). The concentration of HS in relation to EYL changed from 0% to 10% by weight. The influence of HS on various areas of membranes: interphase water-lipid, in the lipid layer just below the polar part of the membrane and in the middle of the lipid bilayer, was investigated by different spin labels (TEMPO, DOXYL 5, DOXYL 16). The study showed that HA slightly decreased the fluidity of the analyzed membranes on the surface layer, while FA significantly liquidated the center of the lipid bilayer. The strong effect of both fractions of HS on the concentration of free radicals as a function of time was also described.

Dynamics of surface of lipid membranes: theoretical considerations and the ESR experiment

The effect of the surface layer of model membranes on their physical properties was discussed in this paper. The research involved a physical ESR experiment with the use of spin probes and computer simulation based on the Monte Carlo technique. Liposomes formed during the process of sonication of lecithin were scanned in an ESR spectrometer. The membrane surface layer model, represented by the system of electric dipoles arranged in rectangular or hexagonal matrices, was studied. The final states of computer simulations were presented as textures. It was found that in the gel phase some ordered domain structures are formed, while in the liquid–crystal phase we got complex textures comprising a plurality of gaps. The process of forming domain structures during the changing of the temperature and the phase transitions taking place in a dipole system as a function of dipole mobility (k-parameter) was presented. The results obtained imply that the head groups (represented by electric dipoles in the computer model) of the surface layer play a key role in membranes, affecting the properties of the entire membrane, which is particularly essential for transport processes. It also modified the characteristics of the membrane gel-liquid crystalline transition phase.

The dynamics of the surface layer of lipid membranes doped by vanadium complex : computer modeling and EPR studies

Abstract Penetration of the liposome membranes doped with vanadium complex formed in the liquid-crystalline phase from egg yolk lecithin (EYL) by the TEMPO (2,2,6,6-tetramethylpiperidine-1-oxyl) spin probes has been investigated. The penetration process was followed by 360 hours at 24°C, using the electron spin resonance (EPR) method. The spectroscopic parameter of the partition (F) of this probe indicated that a maximum rigidity of the membrane was at 3% concentration of the vanadium complex. Computer simulations showed that the increase in the rigidity of the membrane corresponds to the closure of gaps in the surface layer of the membrane, and indicates the essential role of the membrane surface in transport processes.

Impact of humic acids on EYL liposome membranes: ESR method

Abstract In this paper, the effects of model (commercial) and natural (extracted from peat) humic substances on the membrane of liposomes formed with egg yolk lecithin (EYL) are presented. In our research, mass concentrations of fulvic and humic acids were used, which in relation to lecithin varied from 0% to 13%. To study membrane fluidity, electron spin resonance (EPR) was used with two spin probes, penetrating various regions of the lipid bilayer. The effects of model and natural humic substances (humic acids – HAs and fulvic acids – FAs) on the lipid membrane in different regions were researched: the lipid-water interphase, and in the middle of the lipid bilayer. It was shown that FA and HA impact the fluidity of liposome membranes in different ways. Increased mass concentrations of HAs decreased membrane fluidity in both acids: extracted from peat and the model. However, increased mass concentration of FAs extracted from peat, decreased membrane fluidity in the surface region, at the same time stiffening the central part of the bilayer. Increasing the concentration of FAs extracted from peat had the opposite effect when compared to model FA. This effect may be related to the complexation of xenobiotics present in the soil environment and their impact on biological membranes.

Brain Biophysics: Perception, Consciousness, Creativity. Brain Computer Interface (BCI)

The paper presents connections between perception, awareness, and creativity from the biophysical point of view. Attention was drawn to human senses’ limitations and their influence on cognition. The role of interfaces connecting brain with computer and particular role of Brain Computer Interface (BCI) are indicated which the authors believe will be the next stage of human brain supporting technology evolution. It will enable the growth of perception, awareness, and creativity, and consequently lead to social development.

The Possibilities of Using BCI Technology in Biomedical Engineering

The paper presents capabilities of building devices dedicated for persons with heavy mobility dysfunction and indicates the role of interfaces connecting brain with computer (Brain Computer Interface, BCI). Impulses coming from closing eyes, clenching teeth, and tongue movement were proposed as optimal in controlling the applications that manage executable systems. A group of electrodes giving a strong electric signal characteristic for the activity were designated and on the basis of conducted research a proposition of a scientific project concerning building of supporting devices for persons with heavy mobility dysfunction was presented.

Virtual Reality Based Simulators for Neurosurgeons - What We Have and What We Hope to Have in the Nearest Future

High levels of manual skills, good visual-motor coordination, excellent imagination and spatial awareness are the main factors determining the success of neurosurgeons. Proficiency in neurosurgical skills used to be acquired through hands-on training in cadaver labs and in real operating theatres under master neurosurgeon supervision. Most recently, virtual reality (VR) and augmented reality (AR) computer simulations have also been considered as tools for education in the neurosurgical training. The authors review existing solutions and present their own concept of a simulator which could become the useful tool for planning, simulation and training of a specific neurosurgical procedure using patient’s imaging data. The benefits of simulator are particularly apparent in the context of neurovascular operations. It is the field in which it is very difficult for young neurosurgeons to gain proficiency because of the lack of experience caused by the competition between microsurgery and endovascular techniques.

The Influence of Natural and Model Forms of Humic Acids on the Dynamic Parameters of Model Membranes

Abstract In this paper the influence of humic acid concentrations extracted from Histosols (HA-A) and their model forms (HA-B) separated from humic substances commercially produced by Carl Roth GmbH + Co.KG on the dynamic properties of liposome membranes was determined. Differences in the quality of the humic acids (HA-A and HA-B) were determined by the 1HNMR and FTIR methods. Liposomes from the sonication of egg yolk lecithin (EYL) in an aqueous solution and synthetic Dipalmitoylphosphatidylcholine (DPPC) were used. Fluidity of liposome membranes was determined by the EPR technique with spin probes (TEMPO, 16DOIXYL). The electrical parameters of membranes were found using a Keithley 6517 electrometer. Our study showed significant differences in the influence of HA-A and HA-B on the membranes. In the bilayer membranes of the liposomes of HA-A admixture there was slightly more stiffening of the interior of the membrane in comparison to HA-B. A similar effect was observed in the surface layer of the liposome membranes. This difference is particularly evident for DPPC liposomes, however, the EYL liposomes admixture with HA-B slightly increased the fluidity of the surface layer. Electrical study confirmed this effect. The study shows that natural and model forms of humic acids differ in their effects on the activity of tested membrane models. The strong differences in the interaction of HA-A and HA-B on parameter F in DPPC liposomes can be result from the transport of humic acids connected to the metal ions inside the membranes (xenobiotics present in the environment).