Impact of an intensive multimodal educative program on behavioral disorders of polyhandicapped patients: A randomized controlled trial.

BACKGROUND: Hospitalized children and young adults with polyhandicap (PLH) often present with behavioral and relational disorders that are mainly related to their difficulties in communicating and interacting with their environments. Educational support is rarely provided to these patients. An intensive multimodal educative program could help in reducing behavioral disorders and in improving the quality of life of healthcare workers, including nurses and auxiliary nurses. METHODS: A multicenter, randomized controlled trial compared the impact of the usual practice of an educative program (1 h a week) to a multimodal intensive educative program (5 h a week) at 12 months. Patients aged 3-25 with PLH defined by the combination of five criteria (motor deficiency, severe-to-profound mental impairment, daily life dependence, restricted mobility, onset of cerebral lesion at younger than 3 years, and at least one behavioral disorder per week [withdrawn behavior, unexplained crying, teeth grinding, self-injury, aggression, stereotypy, or merycism]) were included. The primary outcome was the evolution of the predominant behavioral disorder between study inclusion and 12 months. Healthcare workers completed questionnaires about chronic stress, coping strategies, and quality of life at study inclusion and at 12 months. RESULTS: Overall, 60 patients were included. Despite a tendency toward reduced teeth grinding, withdrawn and self-injury behaviors, the intervention was not significantly effective: The median duration of continuous behavioral disorders (stereotypy, unexplained crying, withdrawn behavior, and teeth grinding) did not differ between groups. The median frequency of the discontinuous behavioral disorders (self-injury) did not differ between groups. Considering each disorder separately, there was a decrease in teeth grinding, self-injury, and autistic-like traits in the intervention group, although it did not reach statistical significance. This study also suggested decreased depersonalization feelings by healthcare workers. CONCLUSION: Although the study did not show a significant reduction in behavioral disorders in patients with PLH, these results encourage further evaluation of educational management, particularly in regard to patients with self-injury and with withdrawn and teeth-grinding behaviors.

Mild synthesis of poly(HEMA)-networks as well-defined nanoparticles in supercritical carbon dioxide.

Free-radical dispersion polymerisation of 2-hydroxyethyl methacrylate was carried out in supercritical carbon dioxide (scCO2) in the presence of stabilisers based on polyethylene oxide (PEO) and poly(heptadecafluorodecyl acrylate) (PFDA). Different architectures of copolymers (random, palm-tree and diblock) were tested for their surface tension, cloud point and as a stabilising agent. The diblock architecture was found to be the best candidate resulting in poly(HEMA) spherical particles with a size of 316 nm. Furthermore, the effect of the CO2-phobic block (PEO) in the diblock architecture was investigated by using three different chain lengths (1000, 2000, 5000 g mol-1). By optimizing the stabiliser composition and structure, mild reaction conditions have been identified allowing us to obtain well-defined spherical cross-linked poly(HEMA) particles with a mean diameter of unprecedented low size (216 nm) at a temperature as low as 35 °C.

Monolayer kinetic model of formation of ß-cyclodextrin-ß-carotene inclusion complex.

The carotenoids are sensitive molecules and their chemical integrity must be preserved from pro-oxidant elements which could affect and decrease their physiological benefits. The encapsulation based on the inclusion of the carotenoids into cage molecules is a promising approach for preserving over time of the intrinsic properties of the carotenoids. It is well known that cyclic oligosaccharide ß-cyclodextrin (CD) as a cage molecule possesses strong inclusion ability to ß-carotene (C) and as a result of the hydrophobic interactions forms an inclusion complex. In the present paper a monolayer kinetic model was established with the notion to extract more information about the influence of the molecular structure and organization to the interfacial interactions between the interacting species as well as about the role of the specific areas, which are often underestimated in previously studied dispersed systems. We developed the monolayer kinetic model for the formation of the inclusion CD-C complex by applying an experimental approach for following the kinetics by means of measuring the decrease of the surface area (ΔA) versus time (t) at constant surface pressure (π) and the decrease of surface pressure (π) versus time (t) at constant surface area (A). We also visualized by AFM the state of the monolayers at the initial and end points of the kinetic process. The values for the degree (d) and constant (Ka) of the association were estimated and compared with those from the studies of dispersed systems.

New trends in encapsulation of liposoluble vitamins.

Liposoluble vitamins (A, D, E, and K) and carotenoids have many benefits on health. They are provided mainly by foods. At pharmacological doses, they can also be used to treat skin diseases, several types of cancer or decrease oxidative stress. These molecules are sensitive to oxidation, thus encapsulation might constitute an appropriate mean to preserve their properties during storage and enhance their physiological potencies. Formulation processes have been adapted for sensitive molecule, limiting their exposure to high temperature, light or oxygen. Each administration pathway, oral, systemic, topical, transdermal and local, requires different particle sizes and release profile. Encapsulation can lead to greater efficiency allowing smaller administration doses thus diminishing potential hypervitaminosis syndrome appearance and side effects. Carrier formulation can be based on vitamin dissolution in lipid media and its stabilization by surfactant mixture, on its entrapment in a matrix or molecular system. Suitability of each type of carrier will be discussed for each pathway.

Enzymatic proteolysis of alpha gliadin monolayer spread at the air-water interface.

The mechanism of the enzymatic hydrolysis under the proteolytic enzyme action of a plant protein alpha gliadin organized as a model monolayer system at the air/water interface was studied. The advantage of the monolayer technique is the ability to control and modify easily the interfacial organization of the molecules and the possibility to optimize the conditions for the hydrolysis. Enzymatic hydrolysis was studied by using a traditional barostat surface balance. The hydrolysis kinetic was followed by measuring simultaneously the decrease of the surface area and change of the surface potential with time. The decrease with time in film area is result of the random scission of the peptide bonds of polypeptide chain and their solubilization in the aqueous subphase. The interpretation of the surface potential data is based on the contribution of the dipole moments of the intact and broken peptide groups. An appropriate kinetic model describing the proteolytic action of a peptidase was applied and the global kinetic constant was obtained. The random scission of the protein chains gave kinetic constants comparable with those measured during the hydrolytic scission of polyester macromolecules but quite different to the values obtained with short-chain lipids.

Self-assembled monolayers of bisphosphonates: influence of side chain steric hindrance.

Bisphosphonates form self-assembled monolayers (SAMs) spontaneously on stainless steel, silicon, and titanium oxidized surfaces. We used contact angle measurements, atomic force microscopy, and X-ray reflectivity analysis to study the formation of SAMs on a model surface of ultraflat titanium (rms = 0.2 nm). The results were extended to standard materials (mechanically polished titanium, stainless steel, and silicon) and showed that water-soluble bisphosphonic perfluoropolyether can easily form SAMs, with 100% surface coverage and a layer thickness of less than 3 nm. Hydrophobic (water contact angle >110 degrees on stainless steel or titanium) and lipophobic (methylene iodide contact angle >105 degrees on titanium) properties are discussed in terms of industrial applications.

1H NMR relaxation studies of protein-polysaccharide mixtures.

NMR water proton relaxation was used to characterize the structure of plant proteins and plant protein-polysaccharide mixtures in aqueous solutions. The method is based on the mobility determination of the water molecules in the biopolymer environment in solutions through relaxation time measurements. Differences of conformation between pea globulin and alpha gliadin seem to control the water molecules mobility in their environment. As deduced from the study of complexes, the electrostatic interactions may also play a major role in the water molecule motions. The phase separation induced under specific conditions seems to promote the translational diffusion of structured water molecules whereas the rotational motion was more restricted.

Interactions between hen egg-white lysozyme, PEG2,000, and PLA50 at the air-water interface.

In this paper, we compared the efficiency of polymer films, made of a poly(ethylene glycol) (PEG2,000)/poly(d,l-lactide) (PLA50) mixture, or a PEG2,000-PLA50 copolymer, to prevent adsorption of a model protein, the hen egg-white lysozyme (HEWL), at the air-water interface. This was achieved by analyzing the surface pressure/surface area curves, and the X-ray reflectivity data of the polymer films spread on a Langmuir trough, obtained in absence or in presence of the protein. For both the mixture and the copolymer, the amount of protein adsorbed at the air-water interface decreases when the density of the polymer surface coverage increases. It was shown that even in a condensed state, the polymer film made by the mixture can not totally prevent HEWL molecules to adsorb and penetrate the polymer mixed film, but however, protein molecules would not be directly exposed to the more hydrophobic phase, i.e. the air phase. It was also shown that the configuration adopted by the copolymer at the interface in its condensed state would prevent adsorption of HEWL molecules for several hours; this would be due in particular to the presence of PEG segments in the interfacial film.

Plant protein-polysaccharide interactions in solutions: application of soft particle analysis and light scattering measurements.

The soft particle analysis theory was applied to plant proteins and polysaccharides in solution, to determine the charge density of these polymers and the depth of the layer accessible by counterions according to pH conditions. In addition to the macromolecule shape characterized by light scattering measurements, these properties are also correlated with the optimum coacervation condition, so as to establish the prevalent plant protein-polysaccharide interactions governing the coacervate formation. Globulin was found to be highly charged and spherically shaped. The best coacervation condition was obtained at the pH value, which corresponds to the protein conformation with a dense and compact accessible layer. On the contrary, for the alpha gliadin, bearing a lower charge, a more extended conformation seems to be more favourable. For the plant proteins studied, the coacervation seems to be controlled by the structure of the counter polyanion used: from our model, it turns out that the rod-like structure of arabic gum observed at acidic pH allows the interaction with plant proteins to form coacervates, contrary to the highly charged and spherical structure of alginate.

Rheological interfacial properties of plant protein-arabic gum coacervates at the oil-water interface.

This study concerns the interfacial properties of the plant proteins-arabic gum coacervates, which are involved in encapsulation processes based on complex coacervation. The results make it possible to deduce the prerequisite characteristics of the protein, which are involved in the coacervate interfacial properties. The influence of pH and concentration on protein interfacial properties was also studied so as to enable us to predict the best conditions to achieve encapsulation. It has been established that, to obtain a good encapsulation yield, the coacervate must show high surface-active properties and its adsorption on the oil droplets must be favored compared to the free protein adsorption. On the other hand, mechanical properties of the interfacial film made of the coacervate, appear to be a key parameter, as reflected by the dilational viscoelasticity measurements. When compared to the properties of the proteins films, an increase of the rigidity of the interfacial film was shown with the coacervates. It was also observed that viscoelastic properties of the coacervate film were strongly reduced, as well as the associated relaxation times. In acidic conditions, the coacervates containing alpha-gliadin are characterized by an interfacial viscoelastic behavior. This behavior reflects the softness of the interfacial film. This viscoelasticity allows also the formation of a continuous layer around the oil droplets to be encapsulated. Drop tensiometry is shown to be a method that could allow the most adapted protein to be selected and the conditions of the coacervation process to be optimized with regard to concentration and pH.

Adsorption kinetics and rheological interfacial properties of plant proteins at the oil-water interface.

Adsorption and rheological properties of plant proteins were determined by means of the dynamic pendant drop technique. The plant protein properties were compared with the interfacial properties of gelatin, which is well-known for its surface-active properties and is commonly used in food and health products. The results showed that alpha gliadins (wheat proteins) and pea globulins have the highest surface active properties at the oil-water interface, even higher than gelatin at the same concentration (weight/volume). After a short time of adsorption, alpha gliadin interfacial behavior is characterized by a pronounced viscoelasticity, which was confirmed with time whereas pea protein interfacial behavior became elastic after a long initial adsorption period. Finally, the behavior of gelatin is very close to the alpha gliadin behavior for the short initial adsorption period, whereas it looks like the behavior of legume seed proteins for longer times of the adsorption kinetics. This study emphasizes the importance of the choice of the proteins and the emulsification time in the encapsulation process, according to the interfacial behavior.

Spectroscopic studies on poly(ethylene glycol)-lysozyme interactions.

In the present paper, different spectroscopic methods were applied to evaluate conformational changes of hen egg-white lysozyme (HEWL) in various solvents and in the presence of poly(ethylene glycol) (PEG). In citrate (0.007M, pH=6), or in Tris (0.1M, pH=7.4), no conformational change of the protein was measured across the range of concentrations tested. In addition, HEWL in ultra-pure water revealed no irreversible conformational change and no activity loss, at least at low concentrations (< or =0.2mg/ml). Whereas PEG can induce a reorganization of water molecules, no change of the secondary and tertiary protein conformations was observed in the presence of PEG. In addition, in the presence of PEG of various molecular weights, no change of enzymatic activity of the HEWL was observed across the range of concentrations tested.

Interfacial properties of adsorbed films made of a PEG2000 and PLA50 mixture or a copolymer at the dichloromethane-water interface.

Adsorption kinetics of films of poly(ethylene glycol) (PEG2000) studied by the dynamic pendant drop method showed that PEG2000 was more tensioactive at the dichloromethane (DCM)-water interface than at the air-water interface. When initially solubilized into DCM, PEG2000 segments would form an adsorbed layer with hydrophobic segments buried into the polymer chains turned toward the organic phase. Compression of this layer, accompanied by viscoelastic effects, led to expulsion of some hydrophilic tails toward the water phase. When initially dissolved in water, adsorption of PEG2000 segments led to an elastic PEG2000 layer organized on both sides of the interface. Results showed that when the PEG2000-PLA50 (poly(D,L-lactide)) copolymer film was adsorbed at the DCM-water interface, it resulted in a mixed layer exclusively turned toward DCM and its rheological properties were governed by PLA50. When adsorption at the DCM-water interface resulted from a physical mixture of PEG2000 and PLA50, rheological properties of the film were influenced by the initial localization of PEG2000 in the bulk phases. In the case of a mixed film formed by the adsorption of PLA50 from DCM and PEG2000 from water, results showed that PEG2000 segments totally pushed those of PLA50 away from the interface and exclusively influenced the behavior of the mixed film.

Influence of some formulation parameters on lysozyme adsorption and on its stability in solution.

According to our results concerning the behavior of lysozyme at interfaces, its secondary structure and its enzymatic activity, successful protein encapsulation would need to maintain a pH value far from the enzyme isoelectric point value during the formulation to reduce, in particular, the adsorption of lysozyme molecules at the created interfaces. Moreover, buffers or salt solution must be used in order to keep intact the native secondary conformation of lysozyme, and preserve its enzymatic activity.

Interfacial properties of amiodarone: the stabilizing effect of phosphate anions.

Amiodarone, a drug used in heart therapy, is poorly soluble in water at room temperature, but forms transparent phases much more concentrated than the critical micellar concentration (CMC), when crystals are heated (above 60 degrees C) in presence of water and cooled down to room temperature. These pseudosolutions were supposed to be made of a complex system of micelles. In order to better understand the effects of pH and ion species on the supramolecular organization of amiodarone, interfacial pressure measurements were performed at the air/water interface on a Langmuir trough. Monolayers spread from chloroformic solutions over non bufferered subphases were insoluble at basic pH (NaOH, pH 10) but soluble at acidic pH (HCl, pH 4). However, a higher ionic strength obtained by adding NaCl (0.15 N) or NaH(2)PO(4) (0.15 N) to the subphase stopped the amiodarone solubilization. On an acidic phosphate subphase (NaH(2)PO(4), pH 4.4, 0.15 N), abnormally high surface pressures (>1 mN/m) were measured for high molecular areas (80-200 Å(2)/molecule) suggesting a supramolecular organization of the surface film. Insoluble monolayers were also obtained when the amiodarone supramolecular pseudosolution was spread on neutral (NaH(2)PO(4), pH 6.25, 0.15 N) or acidic (NaH(2)PO(4), pH 4.4, 0.15 N) subphases. However, a great instability on basic subphase (phosphate buffer pH 8.8) indicated the breakage of the supramolecular structure during spreading. These results are discussed taking into account the amiodarone state of ionization and the electrostatic interactions with counterions. Combining the use of phosphate counterions and that of acidic pH opens new perspectives in the optimization of amiodarone intravenous formulations.

Direct qualitative and quantitative characterization of a radiosensitizer, 5-iodo-2'-deoxyuridine within biodegradable polymeric microspheres by FT-Raman spectroscopy.

Non-destructive qualitative and quantitative characterization of a radiosensitizer, 5-iodo-2'-deoxyuridine (IdUrd), incorporated within injectable microspheres of a biodegradable polymer, poly(D,L-lactide-co-glycolide) (PLGA), was performed using Fourier transform (FT) Raman spectroscopy. Raman spectra of IdUrd, free and entrapped in microspheres, were recorded under fluorescence-free conditions, described and assigned. For the Raman bands of the PLGA microspheres, assignments with preferential localization of the corresponding vibrations at lactic or glycolic units were proposed. No evidence for drug-polymer interactions in microspheres was found. This allowed the FT-Raman spectra to be used for the quantification of the IdUrd content in the samples. For the microspheres with IdUrd loadings varying from 2 to 27% of the total weight, the methodology used provided good reproducibility and precision (1%). Within the sensitivity of the technique, samples exposed to sterilization doses (27 kGy) of gamma-radiation did not exhibit marked changes in the drug structure.

Structural Properties of Asymmetric Mixed-Chain Phosphatidylethanolamine Films.

By use of Langmuir isotherms and atomic force microscopy (AFM) experiments, we compare the structural properties of an asymmetric 1-palmitoyl-2-oleoyl-glycero-3-phosphoethanolamine (PE 181/160) film sampled at 21 degrees C on mica plates with those of a film composed of a mixture of the related saturated phospholipid (PE 160/160) and unsaturated phospholipid (PE 181/181). From PE 181/160, the Pi-A curve obtained presents a plateau (35 mN/m) where AFM images allowed us to determine a nucleation process, which governs the growth of condensed domains. This surface transition is interpreted as reflecting hydrophobic interactions mainly between the alkyl chains of the molecules. Moreover Pi-A curves of a mixture (v/v) of the two related saturated and unsaturated phospholipids (PE 160/160 and PE 181/181) show a "double plateau" (50 and 54 mN/m) and we have characterized a partial surface demixing at every surface pressure. Finally we show that only PE 181/160 is able to form bilayers above the collapse pressure. Copyright 1999 Academic Press.

Why does PEG 400 co-encapsulation improve NGF stability and release from PLGA biodegradable microspheres?

PURPOSE: The aim of this work was to understand the mechanism by which co-encapsulated PEG 400 improved the stability of NGF and allowed a continuous release from PLGA 37.5/25 microspheres. METHODS: Microparticles were prepared according to the double emulsion method. PEG 400 was added with NGF in the internal aqueous phase (PEG/PLGA ratio 1/1 and 1.8/1). Its effect was investigated through interfacial tension studies. Protein stability was assessed by ELISA. RESULTS: A novel application of PEG in protein stabilization during encapsulation was evidenced by adsorption kinetics studies. PEG 400 limited the penetration of NGF in the interfacial film of the primary emulsion. Consequently, it stabilized the NGF by reducing the contact with the organic phase. In addition, it avoided the NGF release profile to level off by limiting the irreversible NGF anchorage in the polymer layers. On the other hand, the amount of active NGF released in the early stages was increased. During microparticle preparation, NaCl could be added in the external aqueous phase to modify the structure of microparticles. This allowed to reduce the initial release rate without affecting the protein stability always encountered in the absence of PEG. CONCLUSIONS: PEG 400 appeared of major interest to achieve a continuous delivery of NGF over seven weeks from biodegradable microparticles prepared by the double emulsion technique.

NGF release from poly(D,L-lactide-co-glycolide) microspheres. Effect of some formulation parameters on encapsulated NGF stability.

Poly(d,l-lactide-co-glycolide) (PLGA 37.5/25 and 25/50) biodegradable microparticles, which allow the locally delivery of a precise amount of a drug by stereotactic injection in the brain, were prepared by a W/O/W emulsion solvent evaporation/extraction method which had been previously optimized. The aim of this work was to study the influence of two formulation parameters (the presence of NaCl in the dispersing phase and the type of PLGA) on the NGF release profiles and NGF stability during microencapsulation. A honey-comb-like structure characterized the internal morphology of the microspheres. The initial burst was attributed to the rapid penetration of the release medium inside the matrix through a network of pores and to the desorption of weakly adsorbed protein from the surface of the internal cavities. The non-release fraction of the encapsulated protein observed after twelve weeks of incubation was accounted for firstly by the adsorption of the released protein on the degrading microparticles and secondly by the entanglement of the encapsulated protein in the polymer chains. The use of sodium chloride in the dispersing phase of the double emulsion markedly reduced the burst effect by making the microparticle morphology more compact. Unfortunately, it induced in parallel a pronounced NGF denaturation. Finally, it appeared that microparticles made from a hydrophilic uncapped PLGA 37.5/25 in the absence of salt, allowed the release of intact NGF at least during the first 24 h as determined by both ELISA and a PC12 cell-based bioassay.

Surface characterization of poly(alpha-hydroxy acid) microspheres prepared by a solvent evaporation/extraction process.

This work constitutes the first attempt to characterize the wettability of poly(alpha-hydroxy acid) (PAHA) microspheres in situ, prepared according to a complex process involving emulsification, solvent evaporation, washing and freeze-drying. The analysis of the flotation profile of the microspheres has allowed us to determine both advancing and receding contact angles at the microsphere/air/water interface and furnished information on the organization of poly(vinyl alcohol) (PVA) and bovine serum albumin (BSA) at the surface of the PAHA coating. By the comparison of contact angles measured from model surfaces obtained by sampling pure PAHA, PVA, BSA and mixed PVA/PAHA monolayers on glass and poly(methyl methacrylate) (PMMA) substrates, it was concluded that the emulsifier (PVA or BSA) was strongly anchored to the surfaces of the microspheres. The use of BSA to formulate the microspheres from a single oil-in-water emulsion led to dry particles having a hydrophobic surface. The unfolding of the hydrophilic segments of the BSA embedded at the surface of the microspheres, following immersion in water, increased the wettability of the microspheres by water. The same qualitative results were obtained when PVA was used to stabilize single emulsions. On the other hand, microspheres formulated from a double water-in-oil-in-water emulsion displayed no modifications of their wettability when immersed in water. This can be explained by the absence of mobility of the hydrophilic segments of the emulsifier which are blocked in the surface or at the subsurface of the polymer matrix.