Joanna Kotyńska

Impedance analysis of phosphatidylcholine membranes modified with valinomycin

The effect of the ion carrier valinomycin on the electrochemical features of the phosphatidylcholine membrane was investigated by electrochemical impedance spectroscopy. Phosphatidylcholine and valinomycin were chosen for the study because they fulfil essential functions in lively organisms. The experimental impedance values obtained in the presence of different amounts of carrier, studied with several potassium ion concentrations, were used for the research ability of valinomycin to form a 1:1 potassium ion complex on the lipid bilayer/electrolyte solution interface. Based on derived mathematical equations, the heterogeneous equilibrium constant (Kh), association rate constant of the complex (kR) and dissociation rate constant of the complex (kD) were calculated. The result of the investigation is the proposal of a new method for the determination of the parameters used to describe the chemical reaction at the interface between a carrier molecule from the membrane and a monovalent ion from the aqueous phase.

The Effect of Contrast Medium SonoVue® on the Electric Charge Density of Blood Cells

The effect of contrast medium SonoVue® on the electric charge density of blood cells (erythrocytes and thrombocytes) was measured using a microelectrophoretic method. We examined the effect of adsorbed H+ and OH− ions on the surface charge of erythrocytes or thrombocytes. Surface charge density values were determined from electrophoretic mobility measurements of blood cells performed at various pH levels. The interaction between solution ions and the erythrocyte’s or thrombocyte’s surface was described by a four-component equilibrium model. The agreement between the experimental and theoretical charge variation curves of the erythrocytes and thrombocytes was good at pH 2–9. The deviation observed at a higher pH may be caused by disregarding interactions between the functional groups of blood cells.

The Effect of Fatal Carbon Monoxide Poisoning on the Surface Charge of Blood Cells

The objective of this investigation was to evaluate postmortem changes of electric charge of human erythrocytes and thrombocytes after fatal carbon monoxide (CO) poisoning. The surface charge density values were determined on the basis of the electrophoretic mobility measurements of the cells carried out at various pH values of electrolyte solution. The surface charge of erythrocyte membranes after fatal CO poisoning as well as after sudden unexpected death increased compared to the control group in the whole range of experimental pH values. Also, a slight shift of the isoelectric point of erythrocyte membranes to high pH values was observed. The surface charge of thrombocyte membranes after fatal CO poisoning decreased at low pH compared to the control group. However, at high pH, the values increased compared to the control group. The isoelectric point of thrombocyte membranes after fatal CO poisoning was considerably shifted toward low pH values compared to the control group. The observed changes are probably connected with the destruction of blood cell structure.

Effect of monovalent ion adsorption on the electric charge of phosphatidylcholine - decylamine liposomal membranes

We examined the effect of adsorbed monovalent ions on the surface charge of phosphatidylcholine (PC) – decylamine (DA) liposomal membranes. Surface charge density values were determined from electrophoretic mobility measurements of lipid vesicles performed at various pH levels. The interaction between solution ions and the PC-DA liposomal surface was described by a six component equilibrium model. The previously determined association constants of the -PO(-) and –N(+)(CH3)3 groups of PC with H+, OH-, Na+ and Cl- ions (KA1H, KB1OH, KA1Na, KB1C1) were used to calculate KB2OH, and KB2C1, the association constants of the –N(+)H3 group of DA with OH- and Cl- ions, providing an experimental verification for the proposed model.

The Effects of Silica Nanoparticles on Apoptosis and Autophagy of Glioblastoma Cell Lines

Silica nanoparticles (SiNPs) are one of the most commonly used nanomaterials in various medical applications. However, possible mechanisms of the toxicity caused by SiNPs remain unclear. The study presented here provides novel information on molecular and cellular effects of SiNPs in glioblastoma LBC3 and LN-18 cells. It has been demonstrated that SiNPs of 7 nm, 5–15 nm and 10–20 nm induce time- and dose-dependent cytotoxicity in LBC3 and LN-18 cell lines. In contrast to glioblastoma cells, we observed only weak reduction in viability of normal skin fibroblasts treated with SiNPs. Furthermore, in LBC3 cells treated with 5–15 nm SiNPs we noticed induction of apoptosis and necrosis, while in LN-18 cells only necrosis. The 5–15 nm SiNPs were also found to cause oxidative stress, a loss in mitochondrial membrane potential, and changes in the ultrastructure of the mitochondria in LBC3 cells. Quantitative real-time PCR results showed that in LBC3 cells the mRNA levels of pro-apoptotic genes Bim, Bax, Puma, and Noxa were significantly upregulated. An increase in activity of caspase-9 in these cells was also observed. Moreover, the activation of SiNP-induced autophagy was demonstrated in LBC3 cells as shown by an increase in LC3-II/LC3-I ratio, the upregulation of Atg5 gene and an increase in AVOs-positive cells. In conclusion, this research provides novel information concerning molecular mechanisms of apoptosis and autophagy in LBC3 cells.

Changes in surface-charge density of blood cells after sudden unexpected death.

The objective of the investigation was evaluation of postmortem changes of electric charge of human erythrocyte and thrombocyte membranes after sudden unexpected death. The surface charge density values were determined on the basis of the electrophoretic mobility measurements of the cells carried out at various pHs of electrolyte solution. The interactions between both erythrocyte and thrombocyte membranes and electrolyte ions were studied. Values of parameters characterizing the membrane—that is, the total surface concentrations of both acidic and basic groups and their association constants with solution ions—were calculated on the basis of a four-equilibria mathematical model. The model was validated by comparison of these values to experimental data. We established that examined electric properties of the cell membranes are affected by sudden unexpected death. Postmortem processes occurring in the cell membranes can lead to disorders of existing equilibria, which in turn result in changes in values of all the above-mentioned parameters.

Adsorption of Monovalent Ions to Phosphatidylcholine-Cardiolipin Membranes

We examined the adsorption of Na+, Cl−, H+, and OH− ions on phosphatidylcholine-cardiolipin liposomal membrane surfaces using both experimental and theoretical approaches. The dependence of the membrane surface charge density on the pH of the electrolyte solution was determined using microelectrophoresis. A mathematical model comprising eight equilibria was formulated to describe the adsorption of the electrolyte ions. Combination of this model with experimental surface charge density values enabled the determination of association constants. From the good agreement between the experimental and theoretical data, we can conclude that the assumed model is compatible with the experimental data.

Microelectrophoretic investigation of the interactions between liposomal membranes formed from a phosphatidylcholine-phosphatidylglycerol mixture and monovalent ions

Abstract.In this paper, we characterized the interactions between two-component liposomal membranes and monovalent electrolyte ions. Liposomes were formed from neutral (phosphatidylcholine) and anionic (phosphatidylglycerol) lipids mixed in various ratios. Microelectrophoresis was used to determine the dependence of the membrane surface charge density on the p H of the electrolyte solution. Changes in the membrane electric charge caused by the adsorption of Na+ , Cl- , H+ , and OH- ions were observed, and the equilibria among these ions and the phosphatidylcholine-phosphatidylglycerol membrane surface were quantified. We proposed a mathematical model for characterizing these equilibria. Using this model, together with experimental data of the membrane surface charge density, we determined association constants characterizing the equilibria. Knowledge of these parameters was necessary to calculate the theoretical curves of the model. We validated the model by curve-fitting the experimental data points to simulated data generated by the model.Graphical abstract

The Effect of Fatal Carbon Monoxide Poisoning on the Equilibria Between Cell Membranes and the Electrolyte Solution

The effect of fatal carbon monoxide poisoning on equilibria between cell membranes and surrounding ions was described using a theoretical four-equilibria model. The model was developed to obtain parameters characterizing the interactions between solution ions and erythrocyte or thrombocyte membrane surface. The parameters are the total surface concentrations of both acidic and basic groups CA, CB and their association constants with solution ions KAH, KBOH. These parameters were used to calculate the theoretical values of surface charge density. The model was validated by comparison of these values to experimental data, which were determined from the electrophoretic mobility measurements of the blood cells. The experimental and theoretical surface charge density values agree at pH 2–8, and at higher pH, the deviation was observed.

Association of alkali metal cations with phosphatidylcholine liposomal membrane surface.

Interactions of alkali metal cations (Li+, Na+, K+, Cs+) with phosphatidylcholine (PC) liposomal membranes were investigated through experimental studies and theoretical considerations. Using a microelectrophoresis technique, charge densities of experimental membrane surfaces were measured as a function of the pH of electrolyte solutions. Equilibria between the PC liposomal membranes and monovalent ions were mathematically analyzed and described quantitatively through a previously proposed theoretical model. Association constants between functional groups of PC and the studied ions were determined and used to define theoretical curves of membrane surface charge density versus pH. Theoretical and experimental data were compared to verify the model. The PC membrane was found to have the highest affinity for lithium ions, among the ions tested.