Francesco Lopez

Vesicle-Templated Layer-by-Layer Assembly for the Production of Nanocapsules

Hollow structures on the submicrometer scale (nm) are obtained with the assembly of polyelectrolytes according to the layer-by-layer (LbL) technique. Following the LbL procedure, polymers alginate and chitosan were alternatively adsorbed on a vesicular template made of didodecyldimethylammonium bromide (DDAB). Evidence for the removal of the vesicular template entrapped in the alginate/chitosan film is presented. The removal of the vesicular template was achieved through interactions between a nonionic surfactant (Triton X100) and the double-chained surfactant forming the vesicles. The application of this approach allowed the production of hollow nanospheres with a mild procedure, avoiding the use of strong acids or other extreme working conditions that can modify the shell integrity. The obtained nanostructures were characterized by means of dynamic light scattering (DLS), the zeta potential, and scanning electron microscopy (SEM). The SEM analysis demonstrated the presence, after the core removal process, of nanocapsules indistinguishable in size and shape from the parent core-shell system. The analysis of the surface charge of the hollow nanocapsules, after the core dissolution, by zeta potential measurements, indicates good aggregate stability. DLS measurements showed that the size of the nanocapsules is on the order of hundreds of nanometers. Moreover, the size of both the core-shell and the hollow particles did not appear to be perturbed by variations in temperature or ionic strength.

Effects of sulfate ions and slightly acidic pH conditions on Cr(VI) adsorption onto silica gelatin composite

The feasibility of utilizing CTAB-silica gelatin composite (C-SGC) to remove hexavalent chromium from aqueous solutions under different conditions was investigated. Removal of chromate was assessed through evaluation of the adsorption kinetics of chromate ions on the composite under equilibrium conditions in the presence of sulfate ions and at a slightly acidic pH condition (pH 5.8). Adsorption competition tests in the presence of sulfate ions showed that Cr(VI) was still effectively adsorbed from aqueous solution regardless of the presence of the competing anions. In fact, the adsorption kinetics performed at different initial chromate concentrations were unaffected by the presence of 100 mg L(-1) sulfate ions (pH 7.5). The equilibrium adsorption data were fitted by Freundlich adsorption isotherms which confirmed that the adsorption efficiency of chromium on the CTAB-silica gelatin composite was unchanged in the presence of sulfate ions. Further, the adsorption process was shown to be pH dependent and more efficient at slightly acidic pH (5.8). These findings demonstrated a high specificity of the CTAB-silica gelatin composite for chromium, and highlight the possibility of using this matrix for efficient removal of chromium from industrial wastewater without the need to eliminate contaminant sulfate ions.

Nucleotides and nucleolipids derivatives interaction effects during multi-lamellar vesicles formation.

In this paper a micellar interface, constituted by the cationic surfactant CTAB, in presence of 1,2-epoxydodecane and nucleotides was used for catanionic multi-lamellar vesicles (MLVs) formation. The micellar solution of CTAB is able to disperse the 1,2 epoxydodecane in the micellar core promoting the reaction of this reagent with the nucleotide attracted by the positive surface charge of the micellar aggregates. The alkylation of AMP and UMP nucleotides leads to the synthesis of nucleolipids. The behaviour of the supramolecular structures formed depends on the starting reagents (AMP, UMP and AMP+UMP) and on the assembly capabilities of the products. In particular nucleotides and nucleotides derivatives interaction effects are evaluated during the multi-lamellar vesicles formation. NMR spectroscopy and UV-vis measurements performed on MLVs showed strong aryl interactions. Interestingly, NMR spectra revealed prevailing stacking interactions between complementary nucleolipids. The assembly of complementary nucleotides affects the course of the reaction during the MLVs formation. Moreover the MLVs supramolecular stability has been tested by means of turbidity and UV-vis measurements. In particular, an enhanced stability has been found in systems prepared with complementary nucleotides confirming that in these systems the self-assembly process is influenced by nucleolipids interactions. Furthermore by following the hypocromic effect during the micellar catalysis, we showed that even in the earlier stages of the reaction significant differences are detectable.

LIGHT-DEPENDENT AND BIOCHEMICAL PROPERTIES OF TWO DIFFERENT BANDS OF BACTERIORHODOPSIN ISOLATED ON PHENYL-SEPHAROSE CL-4B

Abstract— We report a detailed description of the light‐dependent and biochemical properties of two different bands of isolated and nearly delipidated bacteriorhodopsin obtained from chromatography on phenyl‐Sepharose CL‐4B. The two bands (BR I and BR II) showed a number of markedly different spectroscopic and biochemical characteristics: different absorption maximums in the dark, different light/dark adaptations, different M decay kinetics, different stabilities, different responses to titration with alkali in the dark and different circular dichroism (CD) spectra. Organic phosphate contents of BR I and BR II were measured; we found that more than 90% of purple membrane organic phosphate was removed in the course of chromatography and that the phospholipid/protein molar ratio was always higher in BR I than in BR II. In many functional aspects (high stability, response to light adaptation, spectral changes in the dark by alkali addition and bilobate CD spectrum) the first band appeared to be similar to the purple membrane. We suggest that the functional differences between the two bands depend on the fact that the first band (BR I) contains mostly bacteriorhodopsin aggregates corresponding to purple membrane trimers, while the second band (BR II) contains only bacteriorhodopsin monomers.

Effect of detergent concentration on the thermal stability of a membrane protein: The case study of bacterial reaction center solubilized by N,N-dimethyldodecylamine-N-oxide

We report on the response of reaction center (RC) from Rhodobacter sphaeroides (an archetype of membrane proteins) to the exposure at high temperature. The RCs have been solubilized in aqueous solution of the detergent N,N-dimethyldodecylamine-N-oxide (LDAO). Changes in the protein conformation have been probed by monitoring the variation in the absorbance of the bacteriochlorine cofactors and modification in the efficiency of energy transfer from tryptophans to cofactors and among the cofactors (through fluorescence measurements). The RC aggregation taking place at high temperature has been investigated by means of dynamic light scattering. Two experimental protocols have been used: (i) isothermal kinetics, in which the time evolution of RC after a sudden increase of the temperature is probed, and (ii) T-scans, in which the RCs are heated at constant rate. The analysis of the results coming from both the experiments indicates that the minimal kinetic scheme requires an equilibrium step and an irreversible process. The irreversible step is characterized by a activation energy of 205+/-14 kJ/mol and is independent from the detergent concentration. Since the temperature dependence of the aggregation rate was found to obey to the same law, the aggregation process is unfolding-limited. On the other hand, the equilibrium process between the native and a partially unfolded conformations was found to be strongly dependent on the detergent concentration. Increasing the LDAO content from 0.025 to 0.5 wt.% decreases the melting temperature from 49 to 42 degrees C. This corresponds to a sizeable (22 kJ/mol at 25 degrees C) destabilization of the native conformation induced by the detergent. The nature of the aggregates formed by the denatured RCs depends on the temperature. For temperature below 60 degrees C compact aggregates are formed while at 60 degrees C the clusters are less dense with a scaling relation between mass and size close to that expected for diffusion-limited aggregation. The aggregate final sizes formed at different temperatures indicate the presence of an even number of proteins suggesting that these clusters are formed by aggregation of dimers.

Specific interactions between nucleolipid doped liposomes and DNA allow a more efficient polynucleotide condensation.

The interactions between cationic liposomes doped with the anionic nucleolipid 1,2-dipalmitoyl-sn-glycero-3-cytidine diphosphate (DP-Cyt) and deoxyribonucleic acid (DNA) were investigated. Toward this goal, new liposomal and lipoplex formulations characterized by the presence of the anionic amphiphile DP-Cyt were proposed. The effects of incorporation of the cytosine functionalized lipid DP-Cyt into the cationic bilayers were analyzed by means of electrophoretic mobility, dynamic light scattering (DLS) and fluorescence spectroscopy techniques. These approaches allowed us to follow the DNA condensation process and to identify specific electrokinetic characteristics of liposome and DNA-liposome complexes formation. Specifically, DP-Cyt liposomes and DNA were shown to form electrically stable or unstable complexes depending on the charge ratio between the phosphate group of DNA and the cationic lipid. Remarkably, a prominent role for DP-Cyt in enhancing the DNA binding capacity on liposomes was demonstrated. Zeta potential experiments performed on systems with different liposomes/DNA ratio showed that the value of the charge neutralization point is a function of the content of the incorporated DP-Cyt. As a whole, our data demonstrate that the association of cationic DP-Cyt doped liposomes with DNA is driven by both electrostatic interaction and additional specific interactions at the polar head level based on the cytidine nucleobase.

Release of small hydrophilic molecules from polyelectrolyte capsules: Effect of the wall thickness.

Polymer nanocapsules assembled on cationic liposomes have been built through the layer-by-layer (LbL) technique. Chitosan and alginate, two biocompatible polyelectrolytes, were used to cover the template, where the Rhodamine B was previously loaded. The multishell formed with the alternate deposition of the polyelectrolytes, according to the principles of the LbL assembly, was supposed to change the permeability of the capsule wall. The thickness of the multishell was seen increasing with the number of layers deposited through the observations with the Transmission Electron Microscope. The permeability of the capsules was studied through Rhodamine B release assays. Nanocapsules with seven layers of polyelectrolytes released the dye slowly compared to the capsules with three or five layers. The Ritger-Peppas model was applied to investigate the release mechanisms and a non-Fickian transport behavior was detected regardless of the number of layers. Values of diffusion coefficients of Rhodamine B through the capsule wall were also calculated.

Mesoscopic Structure in Mixtures of Water and 1-Butyl-3-methyl imidazolium tetrafluoborate: A Multinuclear NMR Study

This work is devoted to highlighting some peculiar aspects of ionic liquid self-aggregation in water, which is still a matter of wide debate. Here, the whole water /1-butyl-3-methyl imidazolium (bmim+) tetrafluoborate (

Impact of antioxidants dispersions on the stability and oxidation of water-in-olive-oil emulsions

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