Activation energy of sulfate ion transport across methylated human erythrocyte membranes.

Activation energy EA of the sulfate ions transport process across human erythrocyte membranes modified by reductive methylation has been measured. It has been found that exhaustive reductive methylation (3 times) with formaldehyde and borohydride inhibits the sulfate-equilibrium exchange, by a maximum of about 40%. However, methylation has no measurable effect on activation energy, since the evaluated EA values for control and methylated cells remain the same within the experimental error range.

Influence of amphiphilic compounds on membranes.

On the basis of Gortel & Grendel (J. Exp. Med., 1925, 41, 439-494) discovery, the importance of the lipid bilayer as an integral and indispensible component of the cell membrane is discussed. In particular, attention focuses on the interaction between membranes and amphiphilic substances. The effect on membranes of quaternary ammonium salts, both in the form of pesticides and oxidants as well as organic compounds of tin and lead are discussed in greater detail.

Studies on the structural properties of porous membranes: measurement of linear dimensions of solutes.

This work is concerned with a novel way of studying the basic structural and transport properties of porous membranes. Formulae have been derived for: the total number (N) of pores in the membrane; and mean square radius (r(k)) of pores and for selective membranes: the mean square radius (r(ka)) of pores impermeable to solutes and mean square radius (r(kb)) of permeable pores. The result of the investigation is the proposal of a new research procedure (osmotic) for the determination of the linear dimensions of solutes.

Role of hydrophobic and hydrophilic interactions of organotin and organolead compounds with model lipid membranes.

The present study was conducted to clarify the mechanism of toxicity of organic compounds using lipid model membranes (liposomes and planar lipid membranes). The compounds studied were trialkyltin and trialkyllead chlorides, dialkyltin dichlorides and some inorganic forms of those metals. Two different (anionic and cationic) detergents were also used in the experiments to change the surface properties of liposomes. As a measure of interaction between the compounds studied and model membranes were the release of liposome bound praseodymium and the change in stability of planar membranes under the influence of those compounds. On the basis of the results obtained it was postulated that the mechanism of interaction between tin- and leadorganics and model lipid membranes is a combination of different factors featuring interacting sides. The most important properties determining the behaviour of organic compounds in the interaction were lipophilicity and polarity of different parts of the organics and the steric arrangement they can take in the medium. On the other hand, the surface potential of the lipid bilayer and the environment of the lipid molecules, that play a significant role in the availability of the lipid bilayer to the organics, were important factors in the interaction.

Antioxidant protection of egg lecithin liposomes during sonication.

When model membranes are prepared by ultrasonic treatment of polyunsaturated phospholipids, radical production can induce a partial degradation of the polyunsaturated fatty acyl chains and the formation of lipid hydroperoxides. A suitable antioxidant employed during liposome preparation is able to protect them against lipid peroxidation. This work contains the results of studies on egg lecithin liposomes with incorporated antioxidants that were supposed to play the protective role mentioned. As it has been shown the antioxidant compounds used ensured a 40-60%, i.e., satisfactory protection of liposomes after 30 min sonication. Possible practical applications are discussed.

ESR study of the liposome membrane physical parameters in the heating-cooling cycles.

Changes of dynamic and structural parameters of egg yolk lecithin (EYL) liposome membranes in the heating-cooling cycles have been studied using the ESR spin probe method. The investigations were conducted in the range of temperatures from -18 degrees C to +60 degrees C. It has been found that in the range of temperatures -15 degrees C to +45 degrees C in both the heating and the cooling run the spectroscopic parameters changed practically along the same curve (reversible changes). However, after exceeding this range of temperatures one of the parameters (partition coefficient of the spin probe 2,2,6,6--tetramethylpiperidine -1-oxyl; TEMPO) changed along a closed curve, showing the phenomenon of thermal hysteresis. In the heating process the TEMPO content in liposome membranes was smaller than this in the cooling process. We assume that during the heating, the lipid molecules of the outer liposome layers dissolve in the aqueous medium. In the cooling process they can aggregate and form new liposomes, what in turn increases the surface of liposome membranes, accessible for the TEMPO probe (active surface).

Hemolytic activity of aminoethyl-dodecanoates.

The effect of new lysosomotropic compounds on red blood cell hemolysis and erythrocyte membrane fluidity has been investigated. In earlier studies it was shown that the compounds inhibit the growth of yeast and plasma membrane H(+)-ATPase activity. The study was performed with eight aminoethyl esters of lauric acid variously substituted at nitrogen atom. Esters of dodecanoic acid were chosen for study because at that chain length dimethylaminoethyl esters showed maximum activity. The hemolytic activity of the substances studied exhibits diversified activity in their interaction with the erythrocyte membrane: they differ in hemolytic activity and affect membrane fluidity differently. Erythrocyte membrane fluidity changes under the effect of those compounds which possess highest hemolytic activity. The hemolytic activity of the aminoesters investigated was found to follow a sequence that depended on basicity (i.e. ability of the protonated form formation) of the compound and its polar head group size. The most active are the compounds that possess not more than four carbon atoms substituted at nitrogen and highest pKa value.

Inhibition of lipid peroxidation in the erythrocyte membrane by quaternary morpholinium salts with antioxidant function.

The antioxidant activity of five new compounds from a group of quaternary ammonium salts with antioxidant function has been investigated. The effect of the compounds on the degree of lipid oxidation in the erythrocyte membrane subjected to UV radiation was studied. It was found that all the salts used decreased oxidation of the lipids in the erythrocyte membrane. The antioxidant activity of the ammonium salts studied increased with their alkyl chain length. Three compounds with the longest alkyl chains were the most active antioxidants, and their antioxidant properties were comparable to those of a flavonoid extracted from hawthorn. The antioxidant effectiveness of the ten-carbon alkyl chain compound was comparable with that of the known antioxidant 3,5-di-t-butyl-4-hydroxytoluene (BHT). The least effective antioxidant studied proved to be the eight-carbon alkyl chain compound. The effect of these compounds on fluidity of the erythrocyte membrane has been studied, and for all an increase of fluidity of the membrane was observed. The changes in erythrocyte ghosts fluidity depended both on concentration and type of compound. A fluorimetric study also indicated that rigidity of the erythrocyte membrane increased with degree of its oxidation, but with antioxidants present membrane rigidity increased less.

Bimodal effect of amphiphilic biocide concentrations on fluidity of lipid membranes.

Using the spin label method (ESR) it has been shown that biologically active, amphiphilic compounds (quaternary ammonium salts (AS) containing polar heads with single and double positive charge caused, at low concentrations, decrease fluidity of liposome membranes formed with egg yolk lecithin (EYL). At higher concentrations an increase in fluidity was observed. With compounds having a single positive charge minimum fluidity of membrane structure occurs in the range of 1 to 3%, with compounds containing double positive charge-in the range of 4-6%. That effect does not depend on polar head size and length of alkyl chains of the AS used. Analysis of the electrostatic interaction between positive charges and dipole system suggest that at low ion concentrations the binding energy of the system increases, while it decreases at high concentrations. For the model presented, maximum of binding energy of the system occurs at 3% of positive monovalent ions and at 6% of positive divalent ions admixed.

Erythrocyte hemolysis by organic tin and lead compounds.

The effect of trialkyllead and trialkyltin on pig erythrocyte hemolysis has been studied and compared. The results of experiments showed that the hemolytic activity of organoleads increases with their hydrophobicity and follows the sequence: triethyllead chloride < tri-n-propyllead chloride < tributyllead chloride. And similarly in the case of organotins: triethyltin chloride < tri-n-propyltin chloride < tributyltin chloride. Comparison of the hemolytic activity of organoleads and organotins indicates that the lead compounds exhibit higher hemolytic activity. The methods of quantum chemistry allowed to determine the maximum electric potential of the ions R3Pb+ and R3Sn+, and suggest a relationship between the potential and toxicity.

Organometallics and quaternary ammonium salts affect calcium ion desorption from lecithin liposome membranes.

The objective of the present work was to compare the effects of groups of tin and lead organometallic compounds and their mixtures with amphiphilic quaternary ammonium salts (QAS) on the process of calcium ion desorption from lecithin liposome membranes, as dependent on the properties of the hydrophilic and hydrophobic parts of QAS. In the investigations the method of radioactive labels was applied. Synergism and antagonism in the action of both groups of compounds were found. The effectiveness of the cooperation depended more on chain length of QAS compounds than on the size and polarity of their hydrophobic parts. The most effective of all compounds studied was a the mixture of benzyldimethylammonium chloride in a mixture with tripropyltin. Since the rate of calcium desorption proved to be a good measure of efficacy of biologically active surfactants, it seems that the conclusions reached in this paper may be useful for choosing compounds which are able to decontaminate the environment polluted with heavy metals.

Different effects of di- and triphenyltin compounds on lipid bilayer dithionite permeabilization.

Phenyltins are chemicals widely used in industry, hence their occurrence in the human environment is frequent and widespread. Such compounds include hydrophobic phenyl rings bonded to positively charged tin. This molecular structure makes them capable of adsorbing onto and penetrating through biological membranes, hence they are potentially hazardous. Two such compounds, diphenyltin and triphenyltin, show different steric constraints when interacting with the lipid bilayer. It has been demonstrated that these compounds are positioned at different locations within model lipid bilayers, causing dissimilarity in their ability to affect membrane properties. In this paper we present a study regarding the ability of these two phenyltins to facilitate the transport of S2O4(-2) ions across the lipid bilayer, evaluated by a fluorescence quenching assay. In concentration range of up-to 60 microM those compounds do not affect lipid bilayer topology, when evaluated by vesicle size distribution. Both phenyltins facilitate the transfer of S2O4(-2) across the model lipid bilayer, but the dependence of dithionite transport on phenyltin concentration is different for both. In principle, above 20 microM triphenyltin is more efficient in transferring ions across the lipid bilayer than diphenyltin.

Protective effect of quaternary piperidinium salts on lipid oxidation in the erythrocyte membrane.

A new series of amphiphilic compounds with incorporated antioxidant functional group has been investigated. Piperidinium bromides, differing in the alkyl chain length (8, 10, 12, 14 and 16 carbon atoms in the chain) were synthesised to protect biological and/or model membranes against peroxidation and following negative consequences. Their antioxidant activity was studied with erythrocytes subjected to UV radiation. The salts used inhibited lipid oxidation in the erythrocyte membrane. The degree of this inhibition depended on the alkyl chain length of the bromide used and increased with increasing alkyl chain length. A comparison of the results obtained for piperidinium bromides with those obtained for the widely used antioxidant 3,5-di-t-butyl-4-hydroxytoluene-(BHT) revealed that only two shortest alkyl chain salts were less efficient than BHT in protecting erythrocyte membranes. A similar comparison with antioxidant efficiency of flavonoids extracted from Rosa rugosa showed that they protected the membranes studied more weakly than the least effective eight-carbon alkyl chain piperidinium bromide. The three compounds of longest alkyl chains were the most active antioxidants. Their activities did not differ significantly.

Influence of triphenyllead chloride on biological and model membranes.

The physiological and hemolytic toxicities of triphenyllead chloride (TPhL) as well as its modifying influence on model lipid membranes were studied. The experiments allowed the determination of TPhL concentrations causing 50% inhibition of growth of Spirodela oligorrhiza, Lemna minor and Salvinia natans (EC50), 100% hemolysis of pig erythrocytes (C100) and destabilization of planar lipid membranes (CC). Also, fluidity of erythrocyte ghosts was measured by fluorescence technique and osmotic sensitivity of erythrocytes to the presence of TPhL. All parameters studied were found to be dependent on pH, of experimental solutions and the concentration of TPhL. Acidic conditions increased EC50 C100 and CC concentrations of TPhL. Fluorescence and osmotic measurements showed that osmotic stability and fluidity decreased with increasing trimethyllead concentration. A possible mechanism of TPhL toxicity is discussed. It is assumed that TPhL is interacting with the lipid phase of the models used. It is also assumed that there may exist various, ionic and nonionic, forms of TPhL as a result of its speciation under different experimental conditions. These species, due to their differentiated lipophilicity, may exert different effects on the model membranes studied.

Influence of some lysosomotropic compounds on calcium ion desorption process from liposome membrane.

The effect of a group of model lysosomotropic compounds on the process of Ca2+ ion desorption from lecithin liposome membranes was studied. The compounds studied were: hydrochlorides of fatty acids 2-dimethylaminoethyl esters (DM-n) for n = 9, 11, 13 and 15 carbon atoms in the fatty acid alkyl chain and methochloride of 2-dimethylaminoethyl laurate (DMS-11). It was found that all the compounds studied caused increased desorption with increasing concentration of the compound. Most effective was the quaternary ammonium salt, DMS-11. Moreover, it was found that the process of Ca2+ desorption from the membrane depended on pH of the medium. Compound DM-11 was more active at pH 8 than at pH 5. The action of DM-n compounds depended on the alkyl chain length, DM-11 and DM-13 being the most active. Apparently free amines penetrate the phospholipid membranes and incorporate into its hydrophobic core causing structural deformations. Hydrochlorides of fatty acids and the quaternary ammonium salt induce desorption of calcium ions mostly as a result of competitive electrostatic interactions. By quantum chemistry, PM3 method, and methods of molecular modelling we established the higher hydrophilicity of the polar head of DM-n series with respect to the polar head of the DMS-n compounds. DM-n compounds possess both acceptor and donor properties for hydrogen bonding while DMS-n are instrumental as acceptors only. It should be noted, that the results obtained in this paper for model membranes are in accordance with those for biological ones.

Electrostatic inhibition of hemolysis induced by organotin compounds.

The effect of cations on the kinetics of hemolysis caused by organotin compounds was studied. The ions used in the investigation diminish or totally inhibit hemolysis of red cells induced by organotin compounds. The degree of inhibition depends both on the kind of ion and the compounds that induce hemolysis. The ions Zn2+, Co2+, and Cd2+ present in the medium at 50 microM concentration totally protect the erythrocytes against hemolysis induced by the compound (C3H7)3SnCl. The study has also shown the monovalent ions K+ and trimethyldodecylammonium bromide are less potent inhibitors of hemolysis than divalent ions, which is not the case for two non-ionic organotin compounds only. The studies performed indicate that hemolysis induced by organotin compounds is inhibited due to electrostatic interaction between the cations selected and erythrocyte membrane.