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Calorimetry, X-ray diffraction, transmission electron microscopy and fluorescence techniques have been used to obtain thermodynamic and structural information on dipalmitoyl phosphatidyl choline (DPPC) liposomes doped by the drug fluconazole. The decrease of the gel to liquid crystalline phase transition temperature, as shown by DSC measurements, indicates that fluconazole imparts higher fluidity to the lipid matrix. X-ray diffraction and freeze fracture results show that the presence of the drug stabilizes the two-dimensional lamellar rippled phase Pβ′, prolonging its range of existence, till to room temperature. Fluorescence data show that fluconazole could increase the polarity of fluorescence probe microenvironment. Thus, all the data ensure that fluconazole affects the lipid membrane properties and support the hypothesis that antifungal fluconazole activity is related not only to its structural characteristics but also to its ability to interact with the lipid bilayer.

The biochemical effect of N-acylethanolamines on trout erythrocytes from Salmo irideus was studied by following their susceptibility to hemolysis and measuring steady-state fluorescence anisotropy of the fluorescent probe, 1,6diphenyl-1,3,5-hexatriene (DPH) on isolated membranes. The hemolytic process was strongly affected by different ethanolamine derivatives. However, only slight changes in fluorescence anisotropy were detected in these models, therefore, it appears that acylethanolamine derivatives do not produce significant variations in the physico-chemical characteristics of the membrane. The localization of these molecules in particular microdomains is hypothesized, thus locally modifying physiological membrane activities.

The effect of tri- n -butyltin acetate and tri- n -butyltin chloride on the thermotropic characteristics of dipalmitoyl phosphatidylcholine liposomes was studied. The study was performed by using differential scanning calorimetry and Fourier transform infrared spectroscopy. The results show that tri- n -butyltin acetate affects to a lower extent the hydrophobic core of membrane with respect tri- n -butyltin chloride. The latter considerably lowers the main lipid phase transition and induces a lipid phase separation which was observed at pesticide/lipid molar ratio ( R ) of 10 -2 . At this pesticide concentration the lipid main phase transition encompasses a range of temperature of about 10°C while at 4 × 10 -2 of R value the width of the main transition becomes narrow. These results suggest that, within the same series of trialkyltin compounds, the anion moiety may play an important role in the organotion toxicity

The effect of atrazine on Ca 2+ induced fusion of cardiolipin(CL) and phosphatidylserine (PS) vesicles is studied by Tb 3+ /dipicolinic acid fluorescence and turbidity measurements. The interaction of herbicide with CL and PS membranes is studied by DPH fluorescence polarization. At low concentrations the pesticide partially inhibits fusion, especially in CL vesicles. Higher concentrations of atrazine decrease inhibition of fusion in CL, while fusion is slightly increased in PS. The Ca 2+ -induced increase of turbidity is not affected by atrazine in both PS and CL aggregation experiments. DPH polarization measurements show a perturbation only of the membrane hydrophobic core of PS, in presence of Ca 2+ . It is hypothesized that this biphasic effect shown by low and high atrazine concentrations on Ca 2+ -induced fusion of vesicles is due to a different localization of the pesticide in the membrane.

Atrazin (2-chloro-4 ethylamino-6-(isopropylamino)-s-triazine) is one of the most widely used herbicides. Fourier transform infrared spectroscopy, differential scanning calorimetry and fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene (DPH) and of its derivative 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH) were used to study the interaction of atrazine with dipalmitoyl phosphatidylcholine liposomes used as a model for biological membranes. The results show that atrazine does not perturb the hydrophobic core of the lipid bilayer and suggest that the herbicide localizes near the glycerol backbone of the lipid.

The effect of N-lauroylethanolamine (N-LEA) and N-oleoylethanolamine (N-OEA) on the thermal behaviour of fully hydrated egg phosphatidylethanolamine (TPE) was investigated by the steady-state fluorescence of 2-dimethylamino-(6-lauroyl)-naphtalene (laurdan) and 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH). The parameter generalised polarisation (GP), calculated by exciting laurdan at 340 and 410 nm, revealed the gel to liquid crystalline lamellar (L alpha) as well as the L alpha to inverse hexagonal (HII) phase transitions of TPE. The L alpha to HII phase transition was not detected in TPE/N-OEA system, probably because of the formation of an intermediate Q224 cubic phase. The formation of Q224 phase in TPE/N-OEA and TPE/N-LEA systems was previously demonstrated by X-ray diffraction, but neither laurdan generalised polarisation nor TMA-DPH steady-state fluorescence anisotropy measurements revealed the presence of this phase. It is suggested that the lack of detection of the cubic phase is probably due to the similarity in dynamic characteristics and hydration levels of phospholipid headgroups in the bilayer and cubic phases.

The influence of N-acylethanolamines with different acyl-chains on the physico-chemical state of neutral phospholipids was investigated using dipalmitoyl phosphatidylcholine (DPPC) multilamellar liposomes. The thermal dependence of steady state fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene (DPH) and its charged derivative 1-(4-trimethylaminophenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH) was recorded. The N-acylethanolamines modified the DPPC phase transition temperature and broadened the transition temperature range in different ways depending on the N-acylethanolamines acyl chain characteristics. Our data suggest that the N-acylethanolamine acyl chain length and unsaturation play an important role in the interaction of these compounds with model membranes. The results show that long-chain-N-acylethanolamines interact largely with DPPC model membranes while a similar effect is not observed for the short ones.

The influence of tri-n-butyltin acetate (TBTA) and tri-n-butyltin chloride (TBTC) on the physico-chemical state of charged and neutral phospholipids was investigated using multilamellar liposomes. The thermal dependence of steady state fluorescence polarization of DPH and its charged derivative TMA-DPH was recorded. The two fungicides lowered DPPC phase transition temperature and broadened the temperature range of the transition in different ways. The effects were concentration-dependent. The results show that TBTC interacts more effectively with DPPC model membranes rather than TBTA. Moreover, TBTC broadens and shifts the main phase transition (Tm) more effectively in DPPC rather than in DMPC liposomes. Below Tm, TBTC decreases fluorescence polarization (P) in all phospholipids used. Above Tm P is almost constant in phospholipids with saturated acyl chains, except for DMPG. In fact, an increase of P is detectable in this lipid as in PLs with unsaturated acyl chains. It is suggested that the effects of TBT on liposomal membranes are dependent on the anion moiety and phospholipids characteristics.

The fluorescence decay of 1-(4-trimethylammonium-phenyl)-6-phenyl-1,3,5-hexatriene (TMA-DPH) was used to study micro-heterogeneity of 1,2-dimyristoyl-3-sn-phosphatidylcholine (DMPC) liposomes and to characterize the effect of phosphatidic acid on the correlation between fluorescence microheterogeneity and membrane permeability. The fluorescence decay, measured using multifrequency phase fluorometry, has been analyzed either by using a model of discrete exponential components or a model of continuous distribution of lifetime values. Both analyses have shown that TMA-DPH decay is characterized by two components: a long one of about 9 ns and a short one of about 5 ns. In the gel phase, at variance with previous DPH studies, the short component was associated with a large fractional intensity. The distributional analysis showed changes of lifetime values and width in correspondence to the calorimetric transitions. The presence of egg phosphatidic acid increased both long lifetime values and distributional width. The use of TMA-DPH as a probe to evaluate membrane heterogeneity using the distributional width is discussed. The effect of phosphatidic acid on the membrane surface and in the hydrophobic core has been related to its structural properties and to its role in water penetration.

Experimental evidence of tryptophan incorporation inside dipalmitoyl-lecithin liposomes is reported. The thermodynamic and structural changes induced in the model membranes by tryptophan were investigated by using electron spin resonance (ESR), X-ray diffraction, calorimetry and fluorescence techniques. The amino acid interacts with the polar region of the membrane at low tryptophan concentrations, and with the hydrophobic interior of the membrane at the higher concentrations. The liposomes-tryptophan reciprocal effects are discussed as an example of perturbation induced by an exogenous molecule that penetrates the lipid bilayer. Moreover, they are relevant in relation to the study of lipid-protein interactions and for the possible use of liposomes as amino acid carriers.