Release Of Carboxyfluorescein Research Articles

Bupivacaine, but not lidocaine, disrupts cardiolipin-containing small biomimetic unilamellar liposomes

Inadvertent intravenous administration of bupivacaine, unlike that of lidocaine, is associated with significant cardiotoxicity. However, the mechanism(s) underlying this phenomenon is uncertain. High concentrations of cardiolipin, an anionic phospholipid, are found in the mitochondria membrane of cardiomyocytes. We hypothesized that bupivacaine, but not lidocaine, interacts avidly with cardiolipin in the mitochondria membrane of cardiomyocytes and alters its integrity thereby accounting, in part, for cardiotoxicity. Accordingly, the purpose of this study was to begin to address this issue by determining the effects of bupivacaine and lidocaine on permeability of cardiolipin-containing biomimetic small unilamellar liposomes. We found that bupivacaine, but not lidocaine, elicited a significant, concentration-dependent increase in carboxyfluorescein release from cardiolipin-containing small unilamellar liposomes (size, 165 nm) composed of egg yolk phosphatidylcholine and cholesterol ( p < 0.05). Both drugs had no significant effects on carboxyfluorescein release from liposomes devoid of cardiolipin ( p > 0.5). Collectively, these data indicate that bupivacaine, but not lidocaine, interacts avidly and selectively with biomimetic small unilamellar liposomes containing cardiolipin and disrupts their integrity. We suggest that these interactions underlie, in part, bupivacaine-induced cardiotoxicity.

New GHK hydrophobic derivatives: Interaction with phospholipid bilayers

Three hydrophobic derivatives of GHK peptide containing either N-terminal hexanoyl, decanoyl or myristoyl acyl moieties were synthesized. The binding of these peptidolipids to phospholipid bilayers as well as their hemolytic activity were determined. Moreover, the influence of these peptidolipids on several physicochemical properties of liposomes was studied. Binding experiments indicate a high affinity of these peptidolipids for lipids ordered in liposomes. Nevertheless, this interaction does not promote the release of entrapped carboxyfluorescein. Experiments carried out by the asymmetric membrane method (NBD-PE/dithionite) and quenching studies (PC-pyrene/KI) indicate that this association has a protective effect suggesting that the hydrophobic moiety inserts in the external part of the bilayer and the peptide chain remains protruding from the surface hindering the entrance or the approach of reactants to it. The microviscosity of DPPC bilayers determined using TMA-DPH as fluorescent marker was not affected by the presence of peptidolipids. Besides, results indicate that myristoyl-GHK produces total hemolysis at 2.5 × 10 −4 M but decanoyl and hexanoyl derivatives at 5 × 10 −4 M induce only 10% of hemolysis.

Intracellular distribution of fluorescent probes delivered by vesicles of different lipidic composition

In order to study mechanisms involved in liposome–cell interaction, this work attempted to assess the influence of vesicle composition on the delivery of liposomal content to Hela cells. In particular, to evaluate pH-sensitive properties and cell interaction of the prepared liposomes, the lipid formulations contained cholesterol (Chol) and they were varied by using phosphatidylcholines with different purity degree: soy lecithin (SL; 80% phosphatidylcholine), a commercial mixture of soy phosphatidylcholine (P90; 90% phosphatidylcholine) or dipalmitoylphosphatidylcholine (DPPC; 99% of purity). A second series of liposomes also contained stearylamine (SA). Dehydration-rehydration vesicles (DRV) were prepared and then sonicated to decrease vesicle size. Vesicle–cell interactions and liposomal uptake were examined by fluorescence microscopy using carboxyfluorescein (CF) and phosphatidylethanolamine-dioleoyl-sulforhodamine B (Rho-PE) as fluorescent markers. Fluorescence dequenching assay was used to study the influence of pH on CF release from the liposomal formulations. Liposome adhesion on the cell surface and internalization were strongly dependent on vesicle bilayer composition. SA vesicles were not endocytosed. DPPC/Chol liposomes were endocytosed but did not release their fluorescent content into the cytosol. SL/Chol and P90/Chol formulations displayed a diffuse cytoplasmic fluorescence of liposomal marker.

Synthesis and characterization of hexa-tailed cryptand based amphiphiles: Spontaneous formation of giant vesicular microcapsule with efficient and long-term dye encapsulation

A cryptand with six secondary amino groups has been derivatized by reacting with acid chlorides of different chain lengths (C 7, C 10 and C 18) to get three cryptand based hexa-tailed neutral amphiphiles (L 1–L 3). The cavity of the cryptand head group accommodates two Cu(II) ion giving another set of three amphiphiles. These amphiphiles and its copper complexes can aggregate spontaneously as giant vesicular microcapsules in 10% ethanolic water medium. In all cases vesicles formed are mostly unilamellar in nature. Vesicular microcapsules prepared from L 1–L 3 can encapsulate hydrophilic dye 5(6)-carboxyfluorescein (CF). Stability of microcapsules, CF encapsulation efficiencies and release rates were dependent on the hydrophobic chain length of the amphiphiles. Results show the permeability of the L 2 and L 3 bilayer is lower than that of phosphatidylcholine vesicles and the loading capacity is ∼3 times greater. Microcapsules have been characterized by optical microscopy, freeze fracture scanning electron microscopy and confocal fluorescence microscopy.

Coating of negatively charged liposomes by polylysine: Drug release study

The present work describes surface coating of carboxyfluorescein(CF)-loaded liposomes with poly- l-lysine (PLL) and liposome membrane permeability. The vesicles were prepared from synthetic or natural lipids. Interaction between PLL and the liposomes leads to the formation of complexes — either single PLL-coated vesicles or vesicle aggregates. Formation of the complexes is strongly affected by PLL/lipid molar ratio and the molecular mass of the PLL chains. Liposome permeability depends strongly on the lipid phase state — vesicles in the solid state retained the entrapped dye for a long time, but continuous CF release was registered for “fluid” vesicles. Crossing the transition temperature leads to intensive dye leakage because of the appearance of leaky interfacial domains between the coexisting solid and liquid phases and also because of a reversible change in the vesicle size upon the solid–liquid state phase transition. PLL coverage does not cause permeabilization of “solid” liposomes, but increases the permeability of “fluid” vesicles. At the same time, the results of differential scanning calorimetry and vesicle fusion suggest that PLL adsorption occurs exclusively on the vesicular surface and that the lipidic organization is not significantly disturbed. Moreover, PLL does not prevent lipid exchange between vesicles induced by temperature change.

Structural and functional characterization of a recombinant sticholysin I (rSt I) from the sea anemone Stichodactyla helianthus

Sticholysins I and II (Sts I and II) are two potent cytolysins from the sea anemone Stichodactyla helianthus. These isoforms present 13 substitutions, with three non-conservative located at the N-terminus. St II is considerably more hemolytic than St I in human red blood cells, a result explained by the smaller number of negatively charged groups present at St II's N-terminus. In the present work, we have obtained a recombinant St I (rSt I), differing from the wild type in a single amino acid residue (E16Q). This pseudo-wild type is structurally similar to St I and shows a similar capacity to interact with and form pores in model membranes. This was assessed by the intrinsic fluorescence increase in the presence of liposomes, their adsorption to bilayers (measured by SPR), their concentration at the air–water interface, their interaction with lipid monolayers and their capacity to promote the release of carboxyfluorescein entrapped in liposomes. In spite of these similarities, rSt I presents a larger hemolytic activity in human red blood cells than St I, being intermediate in activity between Sts I and II. The results obtained in the present work emphasize that even the change of one single E by Q at the N-terminal segment may modify the toxin HA and show that this functional property is the most sensitive to subtle changes in the protein primary structure.

A Cost-Effective Method for the Optical Transduction of Chemical Reactions. Application to Hyaluronidase Inhibitor Screening with Polyarginine−Counteranion Complexes in Lipid Bilayers

We report the systematic elaboration of a cost-effective, interference-minimized assay for the label-free optical transduction of chemical reactions. Recently, we have found that certain complexes formed by arginine-rich cell-penetrating peptides (CPPs) and amphiphilic counteranions can act as synergistic anion carriers in lipid bilayer membranes. Application of this discovery to rapid and reversible cytosolic CPP delivery has been described (Futaki, S.; et al. ACS Chem. Biol. 2006, 1, 299). Here, we report the complementary use of polyarginine (pR)-counteranion complexes as general optical transducers of chemical reactions. Counterion screening revealed dodecyl phosphate (DP) as an ideal pR activator. Carboxyfluorescein (CF)-loaded vesicles with a shelf life of 3.5 years served best for the detection of fluorogenic CF release by pR-DP complexes with the naked eye. Inactivation of pR-DP complexes by counterion exchange with hyaluronan (HA) caused no CF emission, while HA removal by hyaluronidase (HAase) did. pR-DP complexes were further compatible with the optical detection of HA immobilization on solid support as well as inhibitor screening for HAase (cromolyn, heparin) with and without substrate immobilization. Controls concerning binary ATP/ADP discrimination for naked-eye kinase detection are mentioned to delineate scope but also limitations of this simple and quite universal method.

Release of FGF1 and p40 synaptotagmin 1 correlates with their membrane destabilizing ability

Fibroblast growth factor (FGF)1 is released from cells as a constituent of a complex that contains the small calcium binding protein S100A13, and the p40 kDa form of synaptotagmin (Syt)1, through an ER–Golgi-independent stress-induced pathway. FGF1 and the other components of its secretory complex are signal peptide-less proteins. We examined their capability to interact with lipid bilayers by studying protein-induced carboxyfluorescein release from liposomes of different phospholipid (pL) compositions. FGF1, p40 Syt1, and S100A13 induced destabilization of liposomes composed of acidic but not of zwitterionic pL. We produced mutants of FGF1 and p40 Syt1, in which specific basic amino acid residues in the regions that bind acidic pL were substituted. The ability of these mutants to induce liposomes destabilization was strongly attenuated, and they exhibited drastically diminished spontaneous and stress-induced release. Apparently, the non-classical release of FGF1 and p40 Syt1 involves destabilization of membranes containing acidic pL.

Dihydroceramide hinders ceramide channel formation: Implications on apoptosis

Early in apoptosis, ceramide levels rise and the mitochondrial outer membrane becomes permeable to small proteins. The self-assembly of ceramide to form channels could be the means by which intermembrane space proteins are released to induce apoptosis. Dihydroceramide desaturase converts dihydroceramide to ceramide. This conversion may be removing an inhibitor as well as generating a pro-apoptotic agent. We report that both long and short chain dihydroceramides inhibit ceramide channel formation in mitochondria. One tenth as much dihydroceramide was sufficient to inhibit the permeabilization of the outer membrane by about 95% (C2) and 51% (C16). Similar quantities inhibited the release of carboxyfluorescein from liposomes indicating that other mitochondrial components are not necessary for the inhibition. The apoptogenic activity of ceramide may thus depend on the ceramide to dihydroceramide ratio resulting in a more abrupt transition from the normal to the apoptotic state when the de novo pathway is used in mitochondria.

Effects of fullerene guests on the stability of 1-palmitoyl-2-oleoylphosphatidylcholine liposomes.

Membrane stability of extruded large unilamellar vesicles () formed by 1-palmitoyl-2-oleoylphosphatidylcholine () containing fullerene C or an amphiphilic fullerene derivative, 2-[2-(2-fulleropyrrolidin-1-ylethoxy)-ethoxy]-ethanol (), has been investigated by spectrofluorimetrically monitoring the spontaneous release of entrapped carboxyfluorescein (). Under controlled conditions of temperature, osmolarity and pH, these guests increase the stability of the liposomal membrane as shown by the decrease in the rate of outflux of . The stability conferred to the liposomes by C and has been compared with that conferred by the well-known stabilizing guest 1,2-dipalmitoyl--glycero-3-phosphoethanolamine--[methoxy-(polyethylene glycol)-2000] ammonium salt (). The addition of amphiphilic molecules, such as non-ionic surfactants, which intercalate into the membrane bilayer, and of sucrose or NaCl, which induce a hyposmotic stress, has been extensively investigated in order to get information on how to modulate the release of entrapped . This information could hopefully be useful in the formulation of new drug delivery systems as well as for getting a deeper understanding of the mechanisms of the formation/enclosure of channels through the membrane. The viscosity and the micropolarity of the membrane have been measured fluorimetrically by using pyrene as the probe. An interesting increase of the gel-liquid crystal phase transition temperature has been observed for POPC liposomes hosting C.

Effect of triorganotin compounds on membrane permeability

Organotin compounds are widely distributed toxicants. They are membrane-active molecules with broad biological toxicity. In this contribution, we study the effect of triorganotin compounds on membrane permeability using phospholipid model membranes and human erythrocytes. Tribultyltin and triphenyltin are able to induce the release of entrapped carboxyfluorescein from large unilamellar vesicles. The rate of release is similar for phosphatidylcholine and phosphatidylserine systems and the presence of equimolar cholesterol decreases the rate of the process. Release of carboxyfluorescein is almost abolished when a non-diffusible anion like gluconate is present in the external medium, and it is restored by addition of chloride. Tributyltin is able to cause hemolysis of human erythrocytes in a dose-dependent manner. Relative kinetics determination shows that potassium leakage occurs simultaneously with hemoglobin release. Hemolysis is reduced when erythrocytes are suspended in a gluconate medium. These results indicate that triorganotin compounds are able to transport organic anions like carboxyfluorescein across phospholipids bilayers by exchange diffusion with chloride and suggest that anion exchange through erythrocyte membrane could be related to the process of hemolysis.

Role of tyrosine-57 and -65 in membrane-damaging and sphingomyelinase activities of Clostridium perfringens alpha-toxin

Clostridium perfringens alpha-toxin (370 residues) is a major virulence factor in the pathogenesis of gas gangrene. The toxin is composed of an N-terminal domain (1-250 residues) where lies the catalytic site and a C-terminal domain (251-370 residues), the Ca(2+)-binding domain, responsible for binding to membranes. The role of Tyr-57 and Tyr-65 close to the catalytic pocket (site) in the N-domain was investigated. Replacement of Tyr-57 and -65 with alanine, leucine, or phenylalanine did not affect the sphingomyelinase activity of the toxin for sodium deoxycholate-solubilized shingomyelin. However, the substitution of Tyr-57 and -65 with alanine or leucine resulted in a radical reduction in the hemolysis of sheep erythrocytes, the release of carboxyfluorescein from shingomyelin-cholesterol (1:1) liposomes, and a significant decrease in binding to the liposomes. The binding of variant toxins, Y57C/C169L and Y65C/C169L, labeled with the environmentally sensitive fluorophore, acrylodan, to the liposomes suggested insertion of the variants in a hydrophobic environment in the bilayer. These observations suggested that Tyr-57 and -65 play a role in the penetration of the toxin into the bilayer of membranes and access of the catalytic site to sphingomyelin in membranes, but do not participate in the enzymatic activity.

A Cellular Deficiency of Gangliosides Causes Hypersensitivity to Clostridium perfringens Phospholipase C

Clostridium perfringens phospholipase C (Cp-PLC), also called alpha-toxin, is the major virulence factor in the pathogenesis of gas gangrene. Previously, a cellular UDP-Glc deficiency was related with a hypersensitivity to the cytotoxic effect of Cp-PLC. Because UDP-Glc is required in the synthesis of proteoglycans, N-linked glycoproteins, and glycosphingolipids, the role of these gly-coconjugates in the cellular sensitivity to Cp-PLC was studied. The cellular sensitivity to Cp-PLC was significantly enhanced by glycosphingolipid synthesis inhibitors, and a mutant cell line deficient in gangliosides was found to be hypersensitive to Cp-PLC. Gangliosides protected hypersensitive cells from the cytotoxic effect of Cp-PLC and prevented its membrane-disrupting effect on artificial membranes. Removal of sialic acids by C. perfringens sialidase increases the sensitivity of cultured cells to Cp-PLC and intramuscular co-injection of C. perfringens sialidase, and Cp-PLC in mice potentiates the myotoxic effect of the latter. This work demonstrated that a reduction in gangliosides renders cells more susceptible to the membrane damage caused by Cp-PLC and revealed a previously unrecognized synergism between Cp-PLC and C. perfringens sialidase, providing new insights toward understanding the pathogenesis of clostridial myonecrosis.

Anion Transport in Liposomes Responds to Variations in the Anchor Chains and the Fourth Amino Acid of Heptapeptide Ion Channels.

Seven heptapeptide derivatives have been prepared. The peptide structure is (Gly)(3)Xxx(Gly)(3) in which Xxx stands for a variable amino acid. The amino acid variations include azetidine carboxylic acid, pipecolic acid, meta-aminobenzoic acid, proline, and leucine. All seven compounds have a C-terminal benzyl group. In all cases, the heptapeptide's N-terminus was linked to diglycolic acid and a dialkylamine. In five cases, the N-terminal group was didecylamine and in two cases, N-ethyl-N-decyl. Chloride and carboxyfluorescein release from phospholipid vesicles was studied with the result that C(10)H(21)N(C(2)H(5)) COCH(2)OCH(2)CO-NH-(Gly)(3)Leu(Gly)(3)-OCH(2)Ph was the most active. Hill analysis showed that this compound involves pore formation by four monomer units rather than two, as previously found for other members of this family.

Evidence for dimer formation by an amphiphilic heptapeptide that mediates chloride and carboxyfluorescein release from liposomes

Heptapeptides having dioctadecyl, N-terminal hydrocarbon chains insert in phospholipid bilayer membranes and form pores through which at least chloride ions pass. Although amphiphilic, these compounds do not typically form vesicles themselves. They insert in the bilayers of phospholipid vesicles and mediate the release of carboxyfluorescein. Hill analysis indicates that at least two molecules of the amphiphile are involved in pore formation. In CD2Cl2, dimer formation is detected by NMR chemical shift changes. The anion release activity of individual anion transporters is increased by linking them covalently at the C-terminus or, even more, by linking them at the N-terminus. Evidence is presented that either linked molecule releases chloride from liposomes more effectively and rapidly than the individual transporter molecule at a comparable concentration.

Cytotoxicity of a GalNAc-Specific C-Type Lectin CEL-I toward Various Cell Lines

We found that CEL-I was a potent cytotoxic lectin. MDCK, HeLa, and XC cells were highly sensitive to CEL-I cytotoxicity and killed in a dose-dependent manner, whereas CHO, L929, and RAW264.7 cells were relatively resistant to CEL-I, and no significant toxicity was observed up to 10 microg/ml. Among these cell lines, MDCK cells showed the highest susceptibility to CEL-I cytotoxicity. A binding study using FITC-labeled CEL-I (F-CEL-I) revealed that the amounts of bound F-CEL-I on the sensitive cell lines were evidently greater than those on the resistant cell lines, suggesting that the different susceptibility of the cell lines to CEL-I cytotoxicity is partly explained by different efficiencies of binding of CEL-I to these cell lines. Interestingly, the cytotoxicity of CEL-I toward MDCK cells was more potent than those of other lectins such as WGA, PHA-L, and Con A, even though these lectins were capable of binding to MDCK cells at comparable levels to CEL-I. Since the cytotoxicity of CEL-I was strongly inhibited by GalNAc, the binding to cell surface specific carbohydrates is essential for the CEL-I cytotoxicity. The trypan blue dye exclusion test indicated that CEL-I caused a disorder of plasma membrane integrity as a relatively early event. CEL-I failed to induce the release of carboxyfluorescein (CF) from CF-loaded MDCK cells as seen for pore-forming hemolytic isolectin CEL-III, suggesting that the primary cellular target of CEL-I may be the plasma membrane, but its action mechanism differs from that of CEL-III. Although CEL-I induced dramatic cellular morphological changes in MDCK cells, neither typical apoptotic nuclear morphological changes nor DNA fragmentation was observed in CEL-I-treated MDCK cells even after such cellular changes. Our results demonstrated that CEL-I showed a potent cytotoxic effect, especially on MDCK cells, by causing plasma membrane disorder without induction of apoptosis.

Biodegradable polymersomes as a basis for artificial cells: encapsulation, release and targeting

The encapsulation of biofunctional compounds, release properties and targetability of polymersomes of amphiphilic block-copolymers based on poly(ethylene glycol) (PEG) and biodegradable polyesters or polycarbonate are described. Carboxyfluorescein (CF), as a model for hydrophilic biofunctional compounds, could be readily incorporated in the polymersomes by adding the compound to the aqueous phase during polymersome preparation. The release of encapsulated material from the polymersomes can be adjusted by changing the copolymer composition, especially the molecular weight and type of hydrophobic block of the copolymer. The presence of plasma proteins other than albumin suppressed the release of CF. CF release in PBS both at room temperature and at 60 °C followed first order kinetics, confirming that the CF containing polymersome system is a membrane controlled reservoir system. These biodegradable polymersomes have the potential to be targeted to specific sites in the body as shown by the specific interaction of anti-human serum albumin immobilized polymersomes with a human serum albumin coated sensor surface.

Biological Activities and Pore Formation of Clostridium perfringens Beta Toxin in HL 60 Cells

Clostridium perfringens beta toxin is an important agent of necrotic enteritis. Of the 10 cell lines tested, only the HL 60 cell line was susceptible to beta toxin. The toxin induced swelling and lysis of the cell. Treatment of the cells with the toxin resulted in K+ efflux from the cells and Ca2+, Na+, and Cl- influxes. These events reached a maximum just before the cells were lysed by the toxin. Incubation of the cells with the toxin showed the formation of toxin complexes of about 191 and 228 kDa, which were localized in the domains that fulfilled the criteria of lipid rafts. The complex of 228 kDa was observed until 30 min after incubation, and only the complex of 191 kDa was remained after 60 min. Treatment of the cells with methyl-beta-cyclodextrin or cholesterol oxidase blocked binding of the toxin to the rafts and the toxin-induced K+ efflux and swelling. The toxin-induced Ca2+ influx and morphological changes were inhibited by an increase in the hydrodynamic diameter of polyethylene glycols from 200 to 400 and markedly or completely inhibited by polyethylene glycol 600 and 1000. However, these polyethylene glycols had no effect on the toxin-induced K+ efflux. The toxin induced carboxyfluorescein release from phosphatidyl-choline-cholesterol liposomes containing carboxyfluorescein and formed an oligomer with 228 kDa in a dose-dependent manner but did not form an oligomer with the 191-kDa complex. We conclude that the toxin acts on HL 60 cells by binding to lipid rafts and forming a functional oligomer with 228 kDa.

Molecular mechanism of membrane permeabilization by the peptide antibiotic surfactin

Surfactin, an acidic lipopeptide produced by various strains of Bacillus subtilis, behaves as a very powerful biosurfactant and possesses several other interesting biological activities. This work deals with the molecular mechanism of membrane permeabilization by incorporation of surfactin. The surfactin-induced vesicle contents leakage was monitored by following release of carboxyfluorescein entrapped into unilamellar vesicles made of palmitoyloleoylphosphatidylcholine (POPC). The effect of the addition of cholesterol, dipalmitoylphosphatidylcholine (DPPC) and palmitoyloleoylphosphatidylethanolamine (POPE) was also checked. It was observed that surfactin was able to induce content leakage at concentrations far below the onset surfactin/lipid ratio for membrane solubilization to occur, which in our system was around 0.92. Electron microscopy showed that vesicles were present after addition of surfactin at a ratio below this value, whereas no vesicles could be observed at ratios above it. Cholesterol and POPE attenuated the membrane-perturbing effect of surfactin, whereas the effect of DPPC was to promote surfactin-induced leakage, indicating that bilayer sensitivity to surfactin increases with the lipid tendency to form lamellar phases, which is in agreement with our previous observation that surfactin destabilizes the inverted-hexagonal structure. Fourier-transform infrared spectroscopy (FTIR) was used to specifically follow the effect of surfactin on different parts of the phospholipid bilayer. The effect on the CO stretching mode of vibration of POPC indicated a strong dehydration induced by surfactin. On the other hand, the CH stretching bands showed that the lipopeptide interacts with the phospholipid acyl chains, resulting in considerable membrane fluidization. The reported effects could be useful to explain surfactin-induced ‘pore’ formation underlying the antibiotic and other important biological actions of this bacterial lipopeptide.

Study on the mechanisms of the antibacterial action of some plant alpha,beta-unsaturated aldehydes.

In this paper the mechanisms involved in the antibacterial effect of six 2E-alkenals [(E)-2-hexenal, (E)-2-eptenal, (E)-2-octenal, (E)-2-nonenal, (E)-2-decenal and (E,E)-2,4-decadienal] were investigated. We measured the release of carboxyfluorescein (CF) trapped in liposomes of phosphatidylcholine (PC) following exposure to the aldehydes mentioned above, in comparison with that elicited by hexanal and nonanal; the modifications of the thermotropic behaviour of liposomes of dimyristoylphosphatidylcholine (DMPC) induced by (E,E)-2,4-decadienal (the aldehyde endowed with the highest microbicidal activity) were evaluated by means of differential scanning calorimetry. With the exception of hexanal, all aldehydes tested caused rapid CF leakage from PC liposomes. The effectiveness order correlates well with the chain length and the presence of the alpha,beta-double bond. Furthermore (E,E)-2,4-decadienal is able to interact with and cross DMPC bilayers. The present findings suggest that the 2E-alkenals tested elicit, very likely, a gross perturbation of the lipidic fraction of plasmatic membranes and are able to penetrate into bacterial cells. These data represent an interesting background for a rational employment of the plant 2E-alkenals tested as antimicrobial agents.