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The use of technologies based on soft polymer particles represents an effective way to deliver target molecules with a specific function. To design a well-performing delivery system, it is fundamental to rationalize both the aggregation and the structural properties of such particles. In this study, we present the kinetic and structural characterization over time of poly(vinyl alcohol) (PVA) microgels obtained through a salting-out process in the presence of NaCl. We have analyzed how both the polymer and salt concentrations affect the aggregation process. The aggregation rate as well as the morphology and physico-chemical parameters, such as mass and chain density of the microgels, have been determined through static and dynamic light scattering and discussed in the framework of the diffusion-limited and reaction-limited colloid aggregation. Insights into the polymer chain arrangements and their dynamics have been gained by means of small-angle neutron scattering and neutron spin-echo measurements. As a result, it was found that NaCl induces a liquid phase separation in solution with the formation of spherical PVA microaggregates, which grow under a reaction-limited aggregation mechanism. The particles increase their size and compactness over time. Within the aggregate, the polymer chains are locally organized to form randomly oriented lamellae with a thickness of about 60 Å. The internal dynamics is a complex mixture of diffusion, Zimm dynamics, and possibly effects from crowding with the transition to a Rouse-like behavior. The microparticle preparation based on the salting-out process constitutes a novelty, if compared to the methods already existing and based on the use of chemical cross-linkers, and is a cheap and easy protocol that allows tuning both particle size and density by varying the salt concentration.

Extracellular polysaccharides are widely produced by bacteria, yeasts, and algae. These polymers are involved in several biological functions, such as bacteria adhesion to surface and biofilm formation, ion sequestering, protection from desiccation, and cryoprotection. The chemical characterization of these polymers is the starting point for obtaining relationships between their structures and their various functions. While this fundamental correlation is well reported and studied for the proteins, for the polysaccharides, this relationship is less intuitive. In this paper, we elucidate the chemical structure and conformational studies of a mannan exopolysaccharide from the permafrost isolated bacterium Psychrobacter arcticus strain 273-4. The mannan from the cold-adapted bacterium was compared with its dephosphorylated derivative and the commercial product from Saccharomyces cerevisiae. Starting from the chemical structure, we explored a new approach to deepen the study of the structure/activity relationship. A pool of physicochemical techniques, ranging from small-angle neutron scattering (SANS) and dynamic and static light scattering (DLS and SLS, respectively) to circular dichroism (CD) and cryo-transmission electron microscopy (cryo-TEM), have been used. Finally, the ice recrystallization inhibition activity of the polysaccharides was explored. The experimental evidence suggests that the mannan exopolysaccharide from P. arcticus bacterium has an efficient interaction with the water molecules, and it is structurally characterized by rigid-rod regions assuming a 14-helix-type conformation.

The formulation of highly concentrated products with low viscosity is a key issue of the surfactants industry, for both economic and ecological reasons. The rational design of surfactant tails bearing a limited number of short side chains has been proposed as a suitable strategy to meet this demand, with negligible effects on biodegradability. In this work, we investigate a mixed surfactant system, in which the branched surfactant N,N-dimethyl-2-propylheptan-1-amine oxide (C10DAO-branched) is combined with a linear alkyl ethoxysulfate one (AES). For comparison, we also study the mixtures in which the branched amine oxide is replaced by its linear isomer (C10DAO-linear). The phase behavior of this surfactant mixture in water is investigated across the entire composition range by polarized optical microscopy and small angle X-ray scattering. Moreover, the shear viscosities and viscoelastic moduli of representative samples are determined by rheological measurements. C10DAO-branched/AES aqueous mixtures form isotropic micellar solution and lamellar structures. Low viscosity was found for all these mixtures, including the most concentrated ones. In contrast, in C10DAO-linear/AES mixtures an extended hexagonal phase is detected, which presents a high viscosity. These results demonstrate tail branching and mixed aggregation to synergistically contribute to the design of high-concentration low-viscosity surfactant mixtures.

Humic acids (HAs) provide an important bio-source for redox-active materials. Their functional chemical groups are responsible for several properties, such as metal ion chelating activity, adsorption ability towards small molecules and antibacterial activity, through reactive oxygen species (ROS) generation. However, the poor selectivity and instability of HAs in solution hinder their application. A promising strategy for overcoming these disadvantages is conjugation with an inorganic phase, which leads to more stable hybrid nanomaterials with tuneable functionalities. In this study, we demonstrate that hybrid humic acid/titanium dioxide nanostructured materials that are prepared via a versatile in situ hydrothermal strategy display promising antibacterial activity against various pathogens and behave as selective sequestering agents of amoxicillin and tetracycline antibiotics from wastewater. A physicochemical investigation in which a combination of techniques were utilized, which included TEM, BET, 13C-CPMAS-NMR, EPR, DLS and SANS, shed light on the structure-property-function relationships of the nanohybrids. The proposed approach traces a technological path for the exploitation of organic biowaste in the design at the molecular scale of multifunctional nanomaterials, which is useful for addressing environmental and health problems that are related to water contamination by antibiotics and pathogens.

Formulations for industrial applications are often mixtures of different surfactants and additives, including salts; thus, it is crucial to understand the reciprocal effects of the different components on the properties of the system such as cmc and morphology of the aggregates in solution. In this respect, we have studied the effects of anions occupying different positions within the so-called Hofmeister series, namely Cl−, Br−, SCN−, on the aggregation properties of mixtures containing a cationic and a non-ionic surfactant largely employed in formulation chemistry: dodecyltrimethyl ammonium bromide (DTAB) and pentaethylenglycol mono octyl ether (C8E5). We determined the cmc of the mixtures with different relative content of the two surfactants, as well as of pure DTAB and C8E5 systems, in water and in the presence of salts. Dynamic light scattering measurements were used to evaluate the hydrodynamic size of the aggregates, while small-angle neutron scattering to determine their morphology, as a function of increasing DTAB content, at constant salt concentration. We found no significant effect of salts on the cmc or aggregate dimensions of C8E5. On the other hand, increasing DTAB content, the salt effect is clearly detectable and ion-dependent: salt addition decreases the cmc in the order SCN− ≫ Br− > Cl−; moreover, halides seem to affect structural properties of mixed C8E5-DTAB systems very slightly whereas SCN− leads to a morphological change from a spherical to a cylindrical aggregate shape. Thus, this study well demonstrates how formulation properties can be fine-tuned by salt addition through a proper choice of ions species.

Hypothesis Catanionic vesicles based on large-scale produced surfactants represent a promising platform for the design of innovative, effective and relatively inexpensive nano-vehicles for a variety of actives. Structural, dynamic and functional behavior of these aggregates is finely tuned by the molecular features of their components and can be opportunely tailored for their applications as drug carriers. Experiments Here we investigate the aggregates formed by CTAC and SDS, two of the most diffused surfactants, by means of Dynamic Light Scattering, Small Angle Neutron Scattering and Electron Paramagnetic Resonance spectroscopy (EPR). The exploitation of these aggregates as nano-vehicles is explored using the poorly water-soluble antioxidant trans -resveratrol ( t -RESV), testing t -RESV solubility and antioxidant activity by means of UV, fluorescence spectroscopy and EPR. Findings The presence of a large stability region of catanionic vesicles on the CTAC-rich side of the phase diagram is highlighted and interpreted in terms of the mismatch between the lengths of the surfactant tails and of first reported effects of the chloride counterions. CTAC-SDS vesicles massively solubilize t -RESV, which in catanionic vesicles exerts a potent antioxidant and radical-scavenging activity. This behavior arises from the positioning of the active at the surface of the vesicular aggregates thus being sufficiently exposed to the external medium.

Background The comprehension of the mechanism of action of antimicrobial peptides is fundamental for the design of new antibiotics. Studies performed looking at the interaction of peptides with bacterial cells offer a faithful picture of what really happens in nature. Methods In this work we focused on the interaction of the peptide Temporin L with E. coli cells, using a variety of biochemical and biophysical techniques that include: functional proteomics, docking, optical microscopy, TEM, DLS, SANS, fluorescence. Results We identified bacterial proteins specifically interacting with the peptides that belong to the divisome machinery; our data suggest that the GTPase FtsZ is the specific peptide target. Docking experiments supported the FtsZ-TL interaction; binding and enzymatic assays using recombinant FtsZ confirmed this hypothesis and revealed a competitive inhibition mechanism. Optical microscopy and TEM measurements demonstrated that, upon incubation with the peptide, bacterial cells are unable to divide forming long necklace-like cell filaments. Dynamic light scattering studies and Small Angle Neutron Scattering experiments performed on treated and untreated bacterial cells, indicated a change at the nanoscale level of the bacterial membrane. Conclusions The peptide temporin L acts by a non-membrane-lytic mechanism of action, inhibiting the divisome machinery. General significance Identification of targets of antimicrobial peptides is pivotal to the tailored design of new antimicrobials.

The ion-specific effect on the solubility of macromolecules in water, rationalized through the Hofmeister series, can be used as an effective method to obtain polymer particles with desired microstructural properties thanks to the so-called salting out effect. By choosing the proper salt and optimizing the polymer and the salt concentration, it is possible to tailor the particle structural features like the radius and the degree of compactness and to speed up the aggregation process until complete conversion from free chains to aggregates. Here we rationalize the segregation of poly-ethylene-vinyl alcohol (EVOH) chains in the presence of sodium chloride (NaCl). The effect of polymer and salt concentrations on the kinetics and the thermodynamics of the aggregation process is considered. In particular, we show how such parameters influence the microstructural properties of the particles through a combination of light and neutron scattering techniques as well as microscopy. We found that by increasing sodium chloride concentration it is possible to speed up the aggregation process, obtaining a complete conversion from free chains to aggregates at shorter times, whereas increasing poly-ethylene-vinyl alcohol concentration from 0.70% w/w to 1.30% w/w determines an increase of the microgel size of a two-times factor. Moreover, NaCl concentration directly affects both the packing degree of the polymer chains within the aggregate and the morphology of the polymer particle, leading to a more compact structure at higher NaCl concentrations. Understanding how it is possible to tune both aggregation kinetics and EVOH particle microstructure is fundamental in order to give a general picture which could be broadened to the class of vinyl alcohol-based polymers.

In the search for more efficient anticancer treatments, Ru(III) complexes have attracted much interest among metal-based candidate drugs, showing marked antitumor and antimetastatic activity associated with lower systemic toxicity. Remarkable examples are the Ru(III) complexes NAMI-A and KP1019, which have reached advanced clinical evaluation. In order to improve the in vivo stability of Ru(III)-based drugs, as well as their cellular uptake and effectiveness, a new approach has been proposed by our research group, based on the incorporation of the active, NAMI-A-like Ru(III) complex into highly functionalized nucleolipidic structures, i.e., hybrid molecules containing a nucleoside or nucleotide central core derivatized with a lipid chain, ensuring both efficient protection against extracellular degradation and high cellular internalization of the metal. Aiming at expanding the chemical diversity of available amphiphilic Ru(III) complexes, we here selected a trifunctional α-amino acid to replace the nucleosidic core of previously prepared nucleolipid-based Ru(III) complexes. The amino acidic scaffold, linked to the Ru(III) complex, is decorated with both hydrophilic and lipophilic moieties, conferring high propensity to form stable aggregates in water, which is required to obtain a suitable nanocarrier for the drug delivery. Following this approach, a novel compound, indicated here as compound I, was successfully prepared and characterized, then studied in coformulation with the biocompatible cationic lipid 1,2-dioleyl-3-trimethylammoniumpropane chloride (DOTAP) by dynamic light scattering (DLS), small angle neutron scattering (SANS), and UV–vis analysis. Evaluated in vitro on a panel of human and nonhuman cell lines, it showed good antiproliferative activity on cancer cells, with IC50 values in the μM range, and no relevant cytotoxicity on the healthy cells used as control.

Several threonine (Thr)- and alanine (Ala)-rich antifreeze glycoproteins (AFGPs) and polysaccharides act in nature as ice recrystallization inhibitors. Among them, the Thr-decorated capsular polysaccharide (CPS) from the cold-adapted Colwellia psychrerythraea 34H bacterium was recently investigated for its cryoprotectant activity. A semisynthetic mimic thereof was here prepared from microbial sourced chondroitin through a four-step strategy, involving a partial protection of the chondroitin polysaccharide as a key step for gaining an unprecedented quantitative amidation of its glucuronic acid units. In-depth NMR and computational analysis suggested a fairly linear conformation for the semisynthetic polysaccharide, for which the antifreeze activity by a quantitative ice recrystallization inhibition assay was measured. We compared the structure-activity relationships for the Thr-derivatized chondroitin and the natural Thr-decorated CPS from C. psychrerythraea. Several threonine (Thr)- and alanine (Ala)-rich antifreeze glycoproteins (AFGPs) and polysaccharides act in nature as ice recrystallization inhibitors. Among them, the Thr-decorated capsular polysaccharide (CPS) from the cold-adapted Colwellia psychrerythraea 34H bacterium was recently investigated for its cryoprotectant activity. A semisynthetic mimic thereof was here prepared from microbial sourced chondroitin through a four-step strategy, involving a partial protection of the chondroitin polysaccharide as a key step for gaining an unprecedented quantitative amidation of its glucuronic acid units. In-depth NMR and computational analysis suggested a fairly linear conformation for the semisynthetic polysaccharide, for which the antifreeze activity by a quantitative ice recrystallization inhibition assay was measured. We compared the structure-activity relationships for the Thr-derivatized chondroitin and the natural Thr-decorated CPS from C. psychrerythraea.

Liposomes are complex aggregates, often including polyethylene glycol (PEG) to expand their life span in vivo, although their full biocompatibility is still questioned. With the aim to suitably replace PEG within liposomal formulations, here we propose a new liposomes formulation, which includes an amphiphilic molecule of natural origin: the lipooligosaccharide (LOS) from the Gram-negative bacterium Rhizobium rubi. LOS architecture is bifunctional: the lipid moiety at one terminus promotes its insertion into the liposome, the other terminus is hydrophilic and in this case presents an oligosaccharide motif similar to the human Lewis B antigen. Liposomes were prepared by co-formulating de-O-acylated LOS (de-LOS) with the lipid 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and the anticancer nucleolipid-based Ru(III) complex, ToThyRu. In-depth microstructural characterization shows that de-LOS containing liposomes are stable aggregates. In vitro preliminary bioscreens have disclosed their negligible toxic profile and a good uptake in MCF-7 and HaCaT cells. The results validate the use of lipooligosaccharides in formulating liposomes and pave the way to their use in drug delivery applications.

Ionophores are an important class of synthetic molecules which mimic natural ion channels or carriers. Here we report the aggregation behavior in pseudo-physiological environment of three Cyclic Phosphate-Linked Oligosaccharides (CyPLOS) derivatives, synthetic ion transporters based on cyclic, phosphate-linked disaccharide skeleton differing for the nature of the tails (tetraethylene–TEG glycol and/or n-undecyl chains) attached to the C-2 and C-3 of the constitutive monosaccharides. Their aggregation behavior has been studied by a combined use of dynamic light scattering (DLS), electron paramagnetic resonance spectroscopy (EPR) and Small Angle Neutron Scattering (SANS). DLS measurements were performed to reveal the formation and size distribution of the CyPLOS aggregates. EPR measurements, by using 5-doxyl stearic acid (5-DSA) as spin-probe, showed that the aggregates are mainly due to the formation of double layers and allowed to analyze the local fluidity. Finally, SANS measurements allowed estimating the layer thickness of the double layers. Our results indicate that the three CyPLOS analogs show self-aggregation properties that depend on the different nature of the inserted tails.

We have experimentally investigated multicomponent diffusion in a protein-polymer-salt-water quaternary system. Specifically, we have measured the nine multicomponent diffusion coefficients, D(ij), for the lysozyme-poly(ethylene glycol)-NaCl-water system at pH 4.5 and 25 degrees C using precision Rayleigh interferometry. Lysozyme is a model protein for protein-crystallization and enzymology studies. We find that the protein diffusion coefficient, D(11), decreases as polymer concentration increases at a given salt concentration. This behavior can be quantitatively related to the corresponding increase in fluid viscosity only at low polymer concentration. However, at high polymer concentration (250 g/L), protein diffusion is enhanced compared to the corresponding viscosity prediction. We also find that a protein concentration gradient induces salt diffusion from high to low protein concentration. This effect increases in the presence of poly(ethylene glycol). Finally, we have evaluated systematic errors associated with measurements of protein diffusion coefficients by dynamic light scattering. This work overall helps characterize protein diffusion in crowded environments and may provide guidance for further theoretical developments in the field of protein crystallization and protein diffusion in such crowded systems, such as the cytoplasm of living cells.

A theoretical spinodal curve for the system water + chloroform + acetic acid at 25 °C is derived using a lattice model for ternary amphiphilic solutions: rod-like molecules covering the bonds of the honeycomb lattice with three-body interactions between the molecular ends associated to the same lattice site. The molecular model is equivalent to the standard Ising model on the same lattice; its mean-field solution is the most appropriate for reproducing, by local fitting, the experimental data for the binodal composition. The derived spinodal curve is in very good agreement with the spinodal composition determined also in the present work from the measured diffusion coefficients recently reported for the same system.

Self-complementary synthetic peptides, composed by 8 and 16 residues, were analyzed by CD, NMR and small angle neutron scattering (SANS) techniques in order to investigate the relevance of charge and hydrophobic interactions in determining their self-assembling properties. All the sequences are potentially able to form fibrils and membranes as they share, with the prototype EAK16, a strictly alternating arrangement of polar and nonpolar residues. We find that 16-mer peptides show higher self-assembling propensities than the 8-mer analogs and that the aggregation processes are favored by salts and neutral pH. Peptide hydrophobic character appears as the most relevant factor in determining self-assembling. Solution conformational analysis, diffusion and SANS measurements all together show that the sequences with a higher self-assemble propensity are distributed, in mild conditions, between light and heavy forms. For some of the systems, the light form is mostly constituted by monomers in a random conformation, while the heavy one is constituted by beta-aggregates. In our study we also verified that sequences designed to adopt extended conformation, when dissolved in alcohol-water mixtures, can easily fold in helix structures. In that media, the prototype of the series appears distributed between helical monomers and beta-aggregates. It is worth noticing that the structural conversion from helical monomer to beta-aggregates, mimics beta-amyloid peptide aggregation mechanisms.

The chemical synthesis by solid-phase methods of a novel amphiphilic peptide, peptide-conjugate amphiphile (PCA), containing in the same molecule three different functions: (i) the N,N-bis[2-[bis(carboxy-ethyl)amino]ethyl]-L-glutamic acid (DTPAGlu) chelating agent, (ii) the CCK8 bioactive peptide, and (iii) a hydrophobic moiety containing four alkyl chains with 18 carbon atoms each, is reported. In water solution at pH 7.4, PCA self-assembles in very stable micelles at very low concentration [critical micellar concentration (cmc) values of 5 x 10(-7) mol kg(-1)] as confirmed by fluorescence spectroscopy. The structural characterization, obtained with small-angle neutron scattering (SANS) measurements, indicates that the aggregates are substantially represented by ellipsoidal micelles with an aggregation number of 39 +/- 2 and the two micellar axes of about 52 and 26 A.

The diffusion coefficients, D ij , for the ternary system water + chloroform + acetic acid are reported at five compositions. This system presents a large solubility gap due to the almost complete insolubility between water and chloroform. The analyzed compositions have a fixed mole ratio of water and chloroform and a decreasing amount of acetic acid when approaching the binodal curve. The difficulty of interpreting the D ij is stressed, and the use of different choices of which component is the solvent and a different reference frame for the diffusive transport is suggested to extract all the possible information from the diffusion coefficients.

Nanostructured hydrogels composed by biocompatible molecules are formulated and characterized. They are based on a polymer network formed by hydrophobically modified chitosans (HMCHIT or CnCHIT) in which vesicles of monoolein (MO) and oleic acid or sodium oleate (NaO), depending on pH , are embedded. The best conditions for gel formation, in terms of pH , length of the hydrophobic moieties of chitosan, and weight proportion among the three components were estimated by visual inspection of a large number of samples. Among all possible combinations, the system C12CHIT–MO–NaO in the weight proportion (1:1:1) is optimal for the formation of a well-structured gel-like system, which is also confirmed by rheological experiments. Electron paramagnetic resonance (EPR) measurements unambiguously show the presence of lipid bilayers in this mixture, indicating that MO–NaO vesicles are stabilized by C12CHIT even at acid pH . A wide small angle neutron scattering investigation performed on several ternary systems of general formula CnCHIT–MO–NaO shows that the length of the hydrophobic tail Cn is a crucial parameter in stabilizing the polymer network in which lipid vesicles are embedded. Structural parameters for the vesicles are determined by using a multilamellar model that admits the possibility of displacement of the center of each shell. The number of shells tends to be reduced by increasing the polymer content. The thickness and the distance between consecutive lamellae are not influenced by either the polymer or MO–NaO concentration. The hydrogel presented in this work, being fully biocompatible and nanostructured, is well-suited for possible application in drug delivery.

A novel nanocomposite system composed of hydrophobically modified ethoxylated urethane (so-called HEUR) polymer, which is a telechelic polymer, and coated magnetite (Fe3O4) nanoparticles (MNp) is investigated. Two different kinds of nanoparticle coatings, are used: a hydrophobic coating composed of oleic acid and oleylamine and a hydrophilic coating having the cationic surfactant C(18)TAB as an additional layer to the hydrophobic nanoparticles. The structure of the nanocomposites as hydrogels with either MNp separately is analyzed using small angle neutron scattering (SANS). Furthermore, the dried state is studied in complementary SANS experiments. The magnetic response of the dried state is characterized by magnometry observing a superparamagnetic behavior.

This chapter presents a summary of manuscripts published in the period of June 2012–June 2013 involving NMR techniques and their applications in the field of amphiphilic systems. In Section 2 reviews and articles on general methods and models have been considered. In Section 3 the manuscripts on thermotropic and lyotropic liquid crystals, phospholipids, vesicles and bicelles have been presented. Section 4 has been devoted to micellar solutions including ionic and non ionic surfactant systems, polymer amphiphiles and mixed amphiphiles systems.

The subject of this article is the combined interpretation of intradiffusion and mutual-diffusion data for polymer–solvent mixtures in terms of integrals over velocity self-correlation functions and velocity cross-correlation functions. The combination of mutual-diffusion, intradiffusion, and activity data allows the evaluation of velocity-correlation coefficients (VCCs) and distinct-diffusion coefficients in systems containing one monodisperse solute. This study is the first attempt to extend these approaches to polymers that are polydisperse solutes. Because of the polydispersity, this correlation analysis may become critical for polymers. Its application to polydisperse samples requires the reduction of intradiffusion and mutual-diffusion coefficients to the same average. After such a reduction, the VCCs and distinct-diffusion coefficients are evaluated for a homologous series of poly(ethylene glycol)s (PEGs). Attractive PEG–PEG interactions depend on the chain length and concentration of PEG. In this analysis, network formation in PEG–water systems appears to be a smooth process. © 2001 John Wiley & Sons, Inc. J Polym Sci Part B: Polym Phys 40: 43–51, 2002

Surface tension, viscosity, intradiffusion, and interdiffusion measurements have been performed on aqueous solutions of sodium octyl sulfonate, sodium decyl sulfonate, and sodium undecyl sulfonate, in the presence of poly(vinylpyrrolidone) at 1% why. The experimental measurements for each ternary system were performed in a range of surfactant concentration including the binary critical micelle concentration of the surfactant. The overall picture of the properties studied suggests the absence of a specific interaction between poly(vinylpyrrolidone) and alkylsulfonate surfactants. The experimental diffusion coefficients can be interpreted in terms of an "excluded volume" effect produced by the presence of the polymer in solution.

Mutual diffusion coefficients were measured with the Gouy technique for two ternary systems: (A) α-cyclodextrin - polyoxyethylene 5 hexylether -water, a non-ionic system, and (B) α-cyclodextrin - sodium 1 hexylsulfonate -water, a ionic system. For both systems the four measured diffusion coefficients allow us to estimate the diffusion coefficients of the actual species present in solution, assuming for the inclusion constant the values obtained from calorimetric data.

Mutual diffusion coefficients have been measured for several average compositions of the system sodium cholate-sodium deoxycholate-water at 25 °C. The experiments have been grouped in different sets having constant concentration of one component and variable concentration of the other one. Following this approach, it has been found that the trends of the main- and cross-term diffusion coefficients can be interpreted on the basis of the diffusion and equilibrium results of similar experiments performed on the two binary systems sodium cholate-water and sodium deoxycholate-water. Implications of the presented results in the transport of lipids operated by bile salt aggregates are mentioned. The method proposed in this work, able to connect the diffusivities of an n-component system to those of the related n-1 subsystems, can be extended to obtain qualitative prediction on the diffusion coefficient trends for mixtures of other surfactants, of both industrial and biological interest.

Aqueous solutions of three surfactants: (1) Sodium 1- hexylsulfonate, (2) Polyoxyethylene - 5 - hexylether, and (3) 1- Hexaminium N,N,N triethyl bromide have been characterized through viscosity, density and calorimetry runs at 25°C. The interaction of these surfactants with α-Cyclodextrin was also studied calorimetrically. The enthalpy of inclusion and the inclusion constant were measured.

Novel thymidine- or uridine-based nucleolipids, containing one hydrophilic oligo(ethylene glycol) chain and one or two oleic acid residues (called ToThy, HoThy and DoHu), have been synthesized with the aim to develop bio-compatible nanocarriers for drug delivery and/or produce pro-drugs. Microstructural characterization of their aggregates has been determined in pure water and in pseudo-physiological conditions through DLS and SANS experiments. In all cases stable vesicles, with mean hydrodynamic radii ranging between 120 nm and 250 nm have been revealed. Biological validation of the nucleolipidic nanocarriers was ensured by evaluation of their toxicological profiles, performed by administration of the nanoaggregates to a panel of different cell lines. ToThy exhibited a weak cytotoxicity and, at high concentration, some ability to interfere with cell viability and/or proliferation. In contrast, DoHu and HoThy exhibited no toxicological relevance, behaving similarly to POPC-based liposomes, widely used for systemic drug delivery. Taken together, these results show nucleolipid-based nanocarriers as finely tunable, multi-functional self-assembling materials of interest for the in vivo transport of biomolecules or drugs.

The present work describes new supramolecular aggregates obtained by co-assembling two different amphiphilic molecules, one containing the bioactive bombesin peptide (BN), or a scramble sequence, and the other, the DOTA chelating agent, (C18)(2)DOTA, capable of forming stable complexes with the radioactive (111)In(III) isotope. The peptide in the amphiphilic monomer is spaced by the lipophilic moiety through ethoxylic spacers of different length: a shorter spacer with five units of dioxoethylene moieties in (C18)(2)L5-peptide, or a longer spacer consisting of a Peg3000 residue in (C18)(2)Peg3000-peptide. Structural characterization by SANS and DLS techniques indicates that, independently from the presence of the peptide containing monomer in the final composition, the predominant aggregates are liposomes of similar shape and size with a hydrodynamic radius R(h) around 200 nm and bilayer thickness, d, of 4 nm. In vitro data show specific binding of the (111)In-(C18)(2)DOTA/(C18)(2)L5-[7-14]BN 90:10 liposomes in receptor expressing cells. However, the presence of the Peg3000 unit on the external liposomal surface, could hide the peptide and prevent the receptor binding. In vivo experiments using (111)In-(C18)(2)DOTA/(C18)(2)L5-[7-14]BN show the expected biological behavior of aggregates of such size and molecular composition, moreover there is an increase in concentration of the GRPR targeting aggregate in the tumors compared to control at the 48 h time point evaluated (2.4% ID/g versus 1.6% ID/g).

A novel fluorescently labelled synthetic ionophore, based on a cyclic phosphate-linked disaccharide (CyPLOS) backbone and decorated with four tetraethylene glycol tails carrying dansyl units, has been synthesised in 12 steps in 26% overall yield. The key intermediate in the synthetic strategy is a novel glucoside building block, serving through its 2- and 3-hydroxy groups as the anchor point for flexible tetraethylene glycol tentacles with reactive azido moieties at their ends. To test the versatility of this glucoside scaffold, it was preliminarily functionalised with a set of diverse probes--as fluorescent, redox-active or hydrophobic tags--either by reduction of the azides followed by condensation with activated carboxylic acid derivatives, or by a direct coupling with a terminal alkyne in a Cu(I)-promoted 1,3-dipolar cycloaddition. Tagging of the monomeric building block with dansyl residues allowed us to prepare a fluorescent, amphiphilic macrocycle, which was investigated for its propensity to self-aggregate in CDCl(3)--studied by means of concentration-dependent (31)P NMR spectroscopy experiments--and in aqueous solution, in which combined dynamic light scattering (DLS) and small-angle neutron scattering (SANS) measurements provided a detailed physico-chemical analysis of the self-assembled systems, mainly organised in the form of large vesicles. Its ion-transport properties through phospholipid bilayers, determined by HPTS fluorescence assays, showed this compound to be more active than the previously synthesised CyPLOS congeners. Solvent-dependent fluorescence changes for the labelled ionophore in liposome suspension established that the dansyl moieties are dispersed in environments with polarity intermediate between those of CH(2)Cl(2) and propan-2-ol, suggesting that the CyPLOS tentacles infiltrate the mid-polar region of the membranes.

The nine mutual diffusion coefficients D ij (i, j=1-3) of the quaternary system α-cyclodextrin (0.02 M)-L-phenylalanine (0.06 M)-monobutylurea (0.06 M)-H 2 O have been determined at 25 o C. Eight reliable experiments were performed at these mean concentrations by the free-diffusion method with a Gouy optical interferometer. A minimum of three experiments is required to obtain the nine diffusion coefficients

New nanocarriers are obtained by assembling two amphiphilic monomers: one containing the bioactive peptide CCK8 spaced, by a polydisperse poly(ethylene glycol), from two hydrophobic tails ((C18)2PEG2000CCK8), and the other containing a chelating agent able to give stable radiolabeled indium-111 complexes linked to the same hydrophobic moiety ((C18)2DTPAGlu). The size and shape of the supramolecular aggregates were structurally characterized by dynamic light scattering, small-angle neutron scattering, and cryogenic transmission electronic microscopy. Under the experimental conditions we investigated (pH 7.4 and molar ratio between monomers 30:70), there is the presence of high polydisperse aggregates: rod-like micelles with a radius of approximately 40 A and length >700 A, open bilayer fragments with thickness approximately 65 A, and probably vesicles. The presence of the bioactive peptide well exposed on the external surface of the aggregate allows selective targeting of nanocarriers towards the cholecystokinin receptors overexpressed by the cancerous cells. In vitro binding assays and in vivo biodistribution studies by nuclear medicine experiments using indium-111 are reported. Moreover, preliminary data concerning the drug loading capability of the aggregates and their drug efficiency on the target cells is reported by using the cytotoxic drug doxorubicin. Incubation of receptor-positive and control cells with peptide-containing aggregates filled with doxorubicin shows significantly lower cell survival in receptor-expressing cells relative to the control, for samples incubated in the presence of doxorubicin.

Diffusion coefficients in the ternary system {alpha}-cyclodextrin (at one concentration)-L-phenylalanine (at four concentrations)-water have been measured by using the Gouy interferometric technique. The effect of the inclusion equilibrium on the cross-term diffusion coefficients was observed. The measured diffusion coefficients in the ternary systems were used to calculate values of the binding constants. These values are in good agreement with the value obtained from calorimetric studies.

Diffusion coefficients in the ternary system α-cyclodextrin (at one concentration)-(DL) norleucine (at three concentrations)-water have been measured using the Gouy interferometric technique. ― The effect of the inclusion equilibrium on the cross-term diffusion coefficients was observed. ― The measured diffusion coefficients in the ternary systems were used to calculate values of the binding constant. These values are in good agreement with that obtained from calorimetric studies

Diffusion, density, and viscosity data are collected for the systems α-cyclodextrin and β-cyclodextrin in water. Frictional coefficients were computed with the help of literature activity data and a qualitative discussion of their concentration dependence was attempted.

Surface tension, fluorescence quenching, and intradiffusion measurements have been performed on sodium decyl sulfonate and sodium decyl sulfate aqueous solutions in the presence and in the absence of poly(vinylpyrrolidone). Despite their chemical similarity, these two surfactants show differences in their behavior even in binary aqueous solution. In fact, the cmc values are different as are their aggregation numbers. Furthermore, the overall picture of the properties we have studied suggests the presence of a preferential interaction of poly (vinylpyrrolidone) toward sodium decyl sulfate with respect to sodium decyl sulfonate, for which no complex formation has been detected. The aqueous poly(vinylpyrrolidone)sodium decyl sulfate system has been extensively studied in order to obtain details on the micellar aggregates bound on the polymer. This includes an estimation of the number of micellelike clusters per polymer molecule and the number of surfactant molecules present in these micelles.

The effect of the polymer molecular weight on the interaction between pentaethylene glycol n-octyl ether (C8E5) and poly(acrylic acid) (PAA) has been investigated by a combined experimental strategy including tensiometry, potentiometry, calorimetry, fluorescence quenching and intradiffusion (pulsed gradient spin echo-NMR) measurements. PAA samples with an average molecular weight varying in a wide range ( M ¯ w = 2000 , 100,000, 250,000, and 450,000) have been considered. The measurements have been performed at constant polymer concentration (0.1% w/w) with varying surfactant molality. In all the considered systems, at low surfactant concentration, adsorption of surfactant monomers onto the polymer chain has been detected. At a C8E5 molality ( T 1 ) independent of the PAA M ¯ w , surfactant molecules start to aggregate, forming clusters to which the polymer co-participates. Above this concentration, the behavior of the system depends on M ¯ w . In fact, if polymer samples with high molecular weight ( M ¯ w ⩾ 100 , 000 ) are employed, all the added surfactant aggregates onto the polymer leading to the polymer saturation and, subsequently, to free micelles formation. Both saturation and free micellization occur at surfactant concentrations which are independent of the polymer molecular weight. C8E5 aqueous mixtures containing PAA with low molecular weight ( M ¯ w = 2000 ) behaves differently, in that, above T 1 , only a fraction ( ∼ 20 % ) of the added surfactant molecules interact with the polymer, forming aggregates to which more than one PAA chain participate. In this case, C8E5 free micellization occurs before polymer saturation. The experimental evidences have been interpreted in terms of the subtle balance between the various molecular interactions driving the surfactant–polymer aggregation.

The relaxivity behaviour and the structural characterization of new supramolecular aggregates (bilayer structures and micelles) obtained by combining two different amphiphilic monomers are reported. One monomer, (C18)(2)DTPAGlu-Gd, contains a very stable gadolinium complex, and the other, DSPE-PEG(2000)-CCK8, contains the bioactive CCK8 peptide. Samples that contained only DSPE-PEG(2000)-CCK8, or up to 50 % (C18)(2)DTPAGlu-Gd, aggregated as double-layer structures (lamellar aggregates) with a thickness of approximately 80-100 A, as evaluated by SANS measurement and Cryo-TEM imaging. A transition to micelle formation was observed when the amount of (C18)(2)DTPAGlu-Gd in the aggregate was increased. These were rod-like micelles approximately 40 A in radius and >200 A in length. The proton relaxivities for both lamellar aggregates and rod-like micelles were the same (17.2 mM(-1) s(-1)), although the values were the results of different combinations of local and global contributions. The in vitro target selectivity of aggregates that contained the CCK-8 peptide was demonstrated by using nuclear medicine techniques.

The structure of chemically-crosslinked chitosan and chitosan-poly(vinylpyrrolidone) (PVP) hydrogels is investigated by means of the combined use of small-angle neutron scattering (SANS), electron paramagnetic resonance spectroscopy (EPR), intradiffusion, and swelling degree measurements. These hydrogels may be described in terms of an inhomogeneous structure composed by polymer-rich and polymer-poor regions. The polymer-rich regions, whose correlation distance zeta is ranged between approximately 600 and approximately 850 A, are, in turn, characterized by the presence of a network formed by the chemical crosslinks, with a mean correlation distance xi approximately 90 A. The structures of chitosan and chitosan-PVP hydrogels have also been analyzed in the presence of sodium decylsulfate micelles that could provide a multidomain system useful, in principle, for drug delivery applications. Both SANS and EPR measurements show that sodium decylsulfate micelles do not significantly interact with both the gels. Finally, intradiffusion and swelling degree measurements show an improved hydrophilicity of chitosan-PVP gels, even further magnified by the presence of C10OS surfactant.

The structure of poly(vinyl alcohol) (PVA) hydrogels formed as a result of freeze/thaw treatments of aqueous solutions of the polymer (11 wt % PVA) in the freshly prepared state is analyzed through the combined use of small (SANS) and ultrasmall (USANS) angle neutron scattering techniques. The structure of these hydrogels may be described in terms of polymer rich regions, with dimensions of the order of 1-2 microm, dispersed in a water rich phase, forming two bicontinuous phases. The PVA chains in the polymer rich phase form a network where the cross-linking points are mainly crystalline aggregates of PVA having average dimensions of approximately 45 A. The structural organization of freeze/thaw PVA hydrogel membranes does not change either after rehydration of dried gels or in the presence of a tensile force. Finally, addition of surfactant micelles inside the gel provides a formulation with both hydrophobic and hydrophilic regions, which demonstrates the potential of the system for drug delivery. Both SANS and EPR measurements show that sodium decylsulfate (C10OS) micelles do not significantly interact with the PVA gel. Variation of the gel structure by the number of freeze/thaw cycles should modulate the rate of release of an active constituent, for example, in a dermal patch.

We report the four diffusion coefficients for the lysozyme-MgCl2-water ternary system at 25 degrees C and pH 4.5. The comparison with previous results for the lysozyme-NaCl-water ternary system is used to examine the effect of salt stoichiometry on the transport properties of lysozyme-salt aqueous mixtures. We find that the two cross-diffusion coefficients are very sensitive to salt stoichiometry. One of the cross-diffusion coefficients is examined in terms of common-ion, excluded-volume, and protein-preferential hydration effects. We use the four ternary diffusion coefficients to extract chemical-potential cross-derivatives and protein-preferential interaction coefficients. These thermodynamic data characterize the protein-salt thermodynamic interactions. We demonstrate the presence of the common-ion effect (Donnan effect) by analyzing the dependence of the preferential-interaction coefficient not only with respect to salt concentration but also with respect to salt stoichiometry. We conclude that the common-ion effect and the protein-preferential hydration are both important for describing the lysozyme-MgCl2 thermodynamic interaction.

In the present work the mixtures water - sodium decylsulfate (C10OS)- poly(vinylpyrrolidone) (PVP) and water - sodium decylsulfonate (C10S) - PVP have been investigated by electron paramagnetic resonance (EPR) spectroscopy employing 4-(N, N-dimethyl-N-(2-hydroxyethyl)) ammonium-2,2,6,6-tetramethyl-piperidine-1-oxyl chloride (TEMPO-choline, TC) as spin probe. TC cations condense on the surface of the aggregates formed by anionic surfactants, acting as counterions, and fit their NO moiety in the outer shell of the hydrophobic core. In water - C10OS - PVP mixtures the nitrogen isotropic hyperfine coupling constant of TC ([A(N)]), reported as a function of the surfactant molality at constant PVP composition, shows two breakpoints: the former ( critical aggregation concentration, c. a. c.) corresponds to the formation of surfactant - polymer clusters while the latter (c(2)) corresponds to the formation of free surfactant micelles. The trend of the correlation time (tau(C)) of the nitroxides in the same system shows that the electrostatic repulsion among the clusters formed onto the PVP macromolecules favours a broadening of the polymer coil and a stiffening of its chain. In contrast, in water - C10S - PVP mixtures no surfactant - polymer interaction could be detected. The effect of the molecular structure of the surfactant headgroup on the surfactant - polymer interaction has been discussed in terms of charge density distribution. In order to further highlight the importance of the headgroup molecular structure on the surfactant association behaviour, the micellisation process of the sodium alkylsulfates (CnOS, n = 6,8,10) has been investigated by the same experimental approach, and the results have been compared with those, previously reported (A. M. Tedeschi, G. D'Errico, E. Busi, R. Basosi and V. Barone, Phys. Chem. Chem. Phys., 2002, 4, 2180, ref. 46), relative to the sodium alkylsulfonates (CnS). CnOS micelles are more ordered and compact with respect to the CnS ones. For the former class of surfactants the electrostatic interaction with TC is stronger and the hydrophobic behaviour of the TC solubilization site is higher.

The interactions in aqueous solution between poly(vinylpyrrolidone) (PVP) and sodium nonanoate (C8COONa), sodium octylsulfonate (C8SNa), lithium perfluorononanoate ( C8FCOOLi), respectively, were investigated. In all the studied systems, the presence of PVP induces a lowering of the aggregation number of the surfactant as evidenced by fluorescence quenching measurements. However, no direct surfactant-polymer interaction has been found for the systems C8COONa PVP and C8SNa PVP. Strong hydrophobic interaction in the system C8FCOOLi PVP has been shown by the fluorescent behavior of pyrene used as a probe. The presence of the fluorinated surfactant induces an increment of similar to14% of the polymer gyration radius, as shown by viscosimetry. Calorimetry confirms the experimental results obtained through fluorescence and shows that the adsorption of C8FCOOLi on PVP is an endothermic process.

The effect of the molecular size of components on preferential solvation analysis has been investigated by studying aqueous solutions of the poly(ethylene glycol) (PEG) homologous chemical series. Precise density data are analyzed for some PEG–water systems, for polymers of molecular weight up to 3400 at 25 °C, at the highest practicable concentrations. The combination of density, activity, and compressibility data allows the evaluation of Kirkwood–Buff integrals for these systems. Data were discussed in terms of preferential solvation, showing that co-operativity takes part to some extent in the PEG–PEG interaction, depending on PEG concentration and molecular weight. The size mismatch between solvent and solute requires a proper reference state, the Matteoli–Shulgin–Ruckenstein state, which takes into account the excluded volume effect. This correction must be applied when considering macromolecular systems, otherwise physically meaningless analyses are developed. Density data at high concentration have been analyzed in order to evaluate progressive aggregation up to network formation, recently proposed for PEG–water systems. The study provides a new picture of aqueous PEG solutions useful for the interpretation of protein precipitation induced by this uncharged polymer. Comparisons with a preferential solvation analysis of the PEG oligomers and with a recent velocity-correlation analysis of PEG polymers have been also made. This paper provides a new picture of PEG–water media, useful to interpret the colloidal stability in the presence of PEG.

The first multicomponent diffusion data ever determined in protein-polymer systems are presented for the system lysozyme(1)-PEG 400(2)-water. Although there are no specific interactions between protein and polymer, the cross-term diffusion coefficient D 21, that links the PEG flow to the protein concentration gradient, is up to 35 times the main-term diffusion coefficient of the protein. This observation can only be due to a "crowding effect" and not to specific interactions, such as electrostatic ones. The exclusion effect is also qualitatively confirmed by the measured counter-flow associated with the protein motion. On the base of a hard core potential, our recent predictive equations are used to predict diffusion coefficients in this ternary system, and a good agreement with the experimental D 21 is obtained. The PEG concentration dependence of the main-term diffusion coefficient of the protein cannot be interpreted exclusively by the excluded volume effect. Some dielectric effect or aggregation phenomena must be invoked to completely describe diffusive behavior in protein-PEG systems. A strong dielectric constant decrease and an anomalous pH dependence on PEG concentration in this system have been observed. We have extended to this nonelectrolyte system a recent procedure for extracting thermodynamic data from ternary diffusion coefficients that uses the Onsager reciprocal relations and the coupling of D ij and second virial coefficient data. Thus we obtained the change of the lysozyme chemical potential with increasing PEG concentration. We emphasize that it is incorrect to neglect the nonideality of PEG-water systems, as was done in some previous preferential solvation analyses.

The multicomponent approach for the description of liquid diffusion processes includes cross-term diffusion coefficients that describe the interaction between the flows of different solute components. We demonstrate that these cross-term diffusion coefficients can be large and thus necessary for the description of diffusion of polydisperse polymers. In this study, the four mutual diffusion coefficients have been measured at 25 °C for the ternary system PEG 2000 (1) [poly(ethylene glycol) 2000 g mol -1] + PEG 200 (2) [poly(ethylene glycol) 200 g mol -1] + water (0). Diffusion coefficients were measured at solute concentrations of 0.0827 mol dm -3 PEG 2000 and 0.076, 0.2293, and 0.4105 mol dm -3 PEG 200. One of the cross-terms, D 21, varies linearly with the concentration of PEG 200 at a constant concentration of PEG 2000 and, at the highest concentration of PEG 200, was larger than either of the main-diffusion coefficients D 11 and D 22. The diffusion coefficients have been qualitatively interpreted in terms of molecular interactions and the excluded volume effect. We also report binary diffusion coefficients, densities, and viscosities for binary solutions of both PEG 200 and PEG 2000 in water at 25 °C.

Isothermal mutual diffusion coefficients (D) have been measured for binary aqueous solutions of sodium alkylsulfonates at 25 degrees C. The diffusion coefficient values drop as the concentration of micelles in the system increases. As the length of the hydrocarbon chain increases, the observed drop in D becomes increasingly sharp and shifts to lower concentrations. When the micellization process is treated as a chemical equilibrium, diffusion coefficients lead to the calculation of the thermodynamic parameters association number, n, and the equilibrium constant, K. These parameters are briefly discussed in connection with the alkyl chain length of the surfactants. Copyright 1999 Academic Press.

Diffusion coefficients have been measured in the ternary system α-cyclodextrin−sodium benzenesulfonate−water at 25 °C. The higher mobility of the inclusion complex, compared to that of free cyclodextrin, proves that in the presence of binding complexes between solutes it is possible to measure positive cross diffusion coefficients. Previously only negative cross terms have been measured in systems containing cyclodextrin−ligand complexes. A discussion of the concentration dependence of the four diffusion coefficients and of the sign of cross terms is given.

The theory of diffusion in n-component systems has been applied to study the mutual diffusion of polydisperse polymeric systems. General relations for n-component systems have been obtained for the experimental Gouy parameters, DA and Q0. Through the use of the Flory equation for diffusion coefficients of monodisperse polymers, the expressions of DA and Q0 at infinite dilution, D∞A and Q∞0, for a general molecular weight distribution have been obtained. These relations have been used to analyze the effect of polydispersity on Gouy diffusion parameters, and the relative information content of Q∞0 is stressed. A procedure is suggested to determine the molecular weight and to estimate the results of the fractionation through the Gouy interferometry. The results of this analysis have been applied to polyethyleneglycol, polyacrylic acid, and sodium polyacrylate diffusion data