Improved biodistribution and enhanced immune response of subunit vaccine using a nanostructure formed by self-assembly of ascorbyl palmitate.

New adjuvant strategies are needed to improve protein-based subunit vaccine immunogenicity. We examined the potential to use nanostructure of 6-O-ascorbyl palmitate to formulate ovalbumin (OVA) protein and an oligodeoxynucleotide (CpG-ODN) (OCC). In mice immunized with a single dose, OCC elicited an OVA-specific immune response superior to OVA/CpG-ODN solution (OC). Rheological studies demonstrated OCC's self-assembling viscoelastic properties. Biodistribution studies indicated that OCC prolonged OVA and CpG-ODN retention at injection site and lymph nodes, reducing systemic spread. Flow-cytometry assays demonstrated that OCC promoted OVA and CpG-ODN co-uptake by Ly6ChiCD11bhiCD11c+ monocytes. OCC and OC induced early IFN-γ in lymph nodes, but OCC led to higher concentration. Conversely, mice immunized with OC showed higher serum IFN-γ concentration compared to those immunized with OCC. In mice immunized with OCC, NK1.1+ cells were the IFN-γ major producers, and IFN-γ was essential for OVA-specific IgG2c switching. These findings illustrate how this nanostructure improves vaccine's response.

Poly(N-isopropylacrylamide)-interpenetrated chitosan coils working as nanoreactors for controlled silver nanoparticle growth.

A new precursor (Ag+/CS/PNIPA) arranged as a nanogel (nanoreactor) is obtained from the aqueous mixture of Ag+, chitosan (CS) and poly(N-isopropylacrylamide) (PNIPA). A model synthetic system based on the thermally induced aqueous silver ions-CS reaction to form silver nanoparticles (AgNP) is used as a starting point to assess the PNIPA role as a thermo-sensitive additive of synthesis in a low content for the production of size-controlled AgNP. As expected, the PNIPA phase transition produced by the temperature increase leads to chitosan nanogel contraction, lowering the diffusion of ionic species. PNIPA behaves as a successful additive between 5.6 and 10.5 wt% of content blended with chitosan, noticeably improving AgNP nucleation during thermal treatment at 90 °C. Higher PNIPA contents are less effective in achieving size control and broader size distributions are generated. The PNIPA effect on the nanoreactor structure is characterized by rheology, modelled and analyzed against the AgNP morphology obtained.

A microfluidic actuator based on a stimuli-responsive hydrogel grafted into Cucurbita moschata xylems.

A chemical actuator was developed taking advantage of the internal microstructure of a plant stem. Stem xylems of Cucurbita moschata were chemically modified with a pH-responsive polymer to obtain an intelligent hydraulic valve. The chemical composition of the device was based mainly on biological scaffolds combined with a minimum content of a tailor-made synthetic copolymer. A pH-sensitive hydrogel composed of a copolymer of acrylic acid was grafted on the inner surface of stem microchannels, assessing the physicochemical properties and the response of the developed actuator under different pressure and pH conditions. Variation of average microcapillary diameter in response to pH stimuli was estimated using Poiseuille's model. This microfluidic device demonstrated the pH-responsive properties and efficient control of flux, showing its open/close transition at pH 3.25 and mechanical stability until pressures of 1.75 meters of water column (mH2O). This actuator has adequate response to open/close cycling and relevance to be evaluated as a pH-response valve of aqueous systems. This kind of actuator is a research topic of high interest with potential application to technology demands.

Physico-chemistry of a successful micro-reactor: Random coils of chitosan backbones used to synthesize size-controlled silver nanoparticles.

The synthetic system to produce silver nanoparticles (AgNP) based on the thermally activated reduction of aqueous silver ions by chitosan (CS) polysaccharide is investigated to unravel the physicochemical processes controlling AgNP nucleation and growth. An anomalous preeminence of AgNP nucleation over growth is found for conditions under which the opposite trend is obeyed for AgNP synthesized from soluble precursors in homogeneous media. The behavior is modeled assuming the formation of a tridimensional supramolecular structure from silver ions / CS´s amino groups coordination complexes, driving the crosslinking within polymer folding and aggregation in shaping random coils. These structures become reactive precursor structures, that behave as microreactors during thermal treatment, and AgNP size are modulated by controlling the amino groups to silver ion ratio. Stabilized AgNP of high quality are easily produced from an environment-friendly synthetic system, which requires low cost reagents and demands simple and safe laboratory procedures.

Enhanced Surface Interaction of Water Confined in Hierarchical Porous Polymers Induced by Hydrogen Bonding.

Hierarchical porous polymer systems are increasingly applied to catalysis, bioengineering, or separation technology because of the versatility provided by the connection of mesopores with percolating macroporous structures. Nuclear magnetic resonance (NMR) is a suitable technique for the study of such systems as it can detect signals stemming from the confined liquid and translate this information into pore size, molecular mobility, and liquid-surface interactions. We focus on the properties of water confined in macroporous polymers of ethylene glycol dimethacrylate and 2-hydroxyethyl methacrylate [poly(EGDMA-co-HEMA)] with different amounts of cross-linkers, in which a substantial variation of hydroxyl groups is achieved. As soft polymer scaffolds may swell upon saturation with determined liquids, the use of NMR is particularly important as it measures the system in its operational state. This study combines different NMR techniques to obtain information on surface interactions of water with hydrophilic polymer chains. A transition from a surface-induced relaxation in which relaxivity depends on the pore size to a regime where the organic pore surface strongly restricts water diffusion is observed. Surface affinities are defined through the molecular residence times near the network surface.

Spatially Resolved Monitoring of Drying of Hierarchical Porous Organic Networks.

Evaporation kinetics of water confined in hierarchal polymeric porous media is studied by low field nuclear magnetic resonance (NMR). Systems synthesized with various degrees of cross-linker density render networks with similar pore sizes but different response when soaked with water. Polymeric networks with low percentage of cross-linker can undergo swelling, which affects the porosity as well as the drying kinetics. The drying process is monitored macroscopically by single-sided NMR, with spatial resolution of 100 µm, while microscopic information is obtained by measurements of spin-spin relaxation times (T2). Transition from a funicular to a pendular regime, where hydraulic connectivity is lost and the capillary flow cannot compensate for the surface evaporation, can be observed from inspection of the water content in different sample layers. Relaxation measurements indicate that even when the larger pore structures are depleted of water, capillary flow occurs through smaller voids.

Evaporation kinetics in swollen porous polymeric networks.

NMR is a fast, nondestructive, and noninvasive technique that can provide information about the pore structure of macroporous polymer beads and the dynamics of liquids confined in them. In this work, we describe the study of the pore structure of the macroporous polymer of ethylene glycol dimethacrylate and 2-hydroxyethyl methacrylate [poly(EGDMA-co-HEMA)] in the dry but also in the swollen state by measuring relaxation times of liquids contained in the polymer network. The results show that the pore architecture differs from the dry to the soaked state. The behavior of polar liquids during evaporation and deswelling dynamics is monitored and described. An internal migration of water from the swollen polymer mesh into expanding pores takes place. With this procedure it is possible to obtain information about the microscopic morphology behavior of the matrix during evaporation and deswelling. This information is of great interest with the aspect of possible and future applications for these types of materials.

Influence of the extraction-purification conditions on final properties of alginates obtained from brown algae (Macrocystis pyrifera).

In this work, three methods (ethanol, HCl, and CaCl(2) routes) of sodium alginate extraction-purification from brown seaweeds (Macrocystis pyrifera) were used in order to study the influence of process conditions on final properties of the polymer. In the CaCl(2) route, was found that the precipitation step in presence of calcium ions followed by proton-exchange in acid medium clearly gives alginates with the lowest molecular weight and poor mechanical properties. It is well known that the acid treatment degrade the ether bonds on the polymeric chain. Ethanol route displayed the best performance, where the highest yield and rheological properties were attained with the lowest number of steps. Although the polymer I.1 showed a molar mass and polydispersity index (M(w)/M(n)) similar to those of commercial sample, its mechanical properties were lower. This performance is related to the higher content of guluronic acid in the commercial alginate, which promotes a more successful calcium chelation. Moreover, the employment of pH 4 in the acid pre-treatment improved the yield of the ethanol route, avoiding the ether linkage hydrolysis. Therefore, samples I.2 and I.3 displayed a higher M(w) and a narrower distribution of molecular weights than commercial sample, which gave a higher viscosity and better viscoelastic properties.

A novel mutation in limb girdle muscular dystrophy.

We describe a patient with limb girdle muscular dystrophy with evidence of a D596N novel mutation of the LMNA gene. He presented with a dilated cardiomyopathy and heart failure. He successfully underwent a cardiac rehabilitation program without cardiovascular complications. Clinicians should suspect a variety of a wide array of diseases including laminopathy, dystrophinopathy, sarcoglynopathy and LGMD 2I. Further studies should focus on determining the specific mode of inheritance and genetic testing should be considered in these patients.

Influence of the morphology of poly(EGDMA-co-HEMA) on the chemical modifications and retention of O-phosphothreonine.

The influence of the morphology of ethylene glycol dimethacrylate-hydroxyethyl methacrylate copolymer [poly(EGDMA-co-HEMA)] base support to obtain different Fe(3+)-containing sorbents and their properties in retention of O-phosphothreonine [Thre(P)] is examined in this paper. Three base supports poly(EGDMA-co-HEMA) (I-III) were obtained using different quantities of initiator in suspension polymerization reactions. These products were submitted to chemical modifications using 1,4-butanediol diglycidyl ether (BDGE) in activation reactions and different chelating agents (iminodiacetic acid, IDA; disodium ethylenediamine tetraacetate, EDTA; and hexamethylenediamine tetrapropanoic acid, HMDTP) in coupling reactions to attain Fe(3+)-containing sorbents. Properties such as specific surface area (S(s)), specific pore volume (V(p)), scanning electron microscopy (SEM), IR spectroscopy, quantity of functional groups (oxirane and carboxyl), amount of chelated metal ion, ligand occupation (L), swelling studies as well as quantity of O-phosphoamino acid retained were used as comparative parameters for matrices. In general, the derivatization reactions proved to be more efficient when higher S(s) of macropores (50-400,000 nm) were available in the matrix. In our case, it was observed when highest percentage of initiator was used. On the other hand, the effect of accessibility of surface area on the yield of coupling reactions was noticed when comparing the different chelating agents since the number of carboxyl groups present in products was higher when the molecular size of the chelating agent was lower. Although all Fe(3+)-containing sorbents resulted efficient to retain Thre(P), the values of retention of the amino acid were slightly higher when IDA-containing matrices were used irrespective of the quantity of metal chelated. This could be probably due to the fact that the IDA ligand could be bounded to the matrix in sites that though accessible for the center of adsorption were hard for Thre(P) to access.