Lata Panicker

Diffusional Transport of Ions in Plasticized Anion-Exchange Membranes

Diffusional transport properties of hydrophobic anion-exchange membranes were studied using the polymer inclusion membrane (PIM). This class of membranes is extensively used in the chemical sensor and membrane based separation processes. The samples of PIM were prepared by physical containment of the trioctylmethylammonium chloride (Aliquat-336) in the plasticized matrix of cellulose triacetate (CTA). The plasticizers 2-nitrophenyl octyl ether, dioctyl phthalate, and tris(2-ethylhexyl)phosphate having different dielectric constant and viscosity were used to vary local environment of the membrane matrix. The morphological structure of the PIM was obtained by atomic force microscopy and transmission electron microscopy (TEM). For TEM, platinum nanoparticles (Pt nps) were formed in the PIM sample. The formation of Pt nps involved in situ reduction of PtCl(6)(2-) ions with BH(4)(-) ions in the membrane matrix. Since both the species are anions, Pt nps thus formed can provide information on spatial distribution of anion-exchanging molecules (Aliquat-336) in the membrane. The glass transitions in the membrane samples were measured to study the effects of plasticizer on physical structure of the membrane. The self-diffusion coefficients (D) of the I(-) ions and water in these membranes were obtained by analyzing the experimentally measured exchange rate profiles of (131)I(-) with (nat)I(-) and tritiated water with H(2)O, respectively, between the membrane and equilibrating solution using an analytical solution of Ficks second law. The values of D(I(-)) in membrane samples with a fixed proportion of CTA, plasticizer, and Aliquat-336 were found to vary significantly depending upon the nature of the plasticizer used. The comparison of values of D with properties of the plasticizers indicated that both dielectric constant and viscosity of the plasticizer affect the self-diffusion mobility of I(-) ions in the membrane. The value of D(I(-)) in the PIM samples did not vary significantly with concentration of Aliquat-336 up to 0.5 mequiv g(-1), and thereafter D(I(-)) increased linearly with Aliquat-336 concentration in the membrane. The self-diffusion coefficients of water D(H(2)O) in PIM samples were found to be 1 order of magnitude higher than the value of D(I(-)) and varied slightly depending upon the plasticizer present in the membrane. It was observed in electrochemical impedance spectroscopic studies of the PIM samples that diffusion mobility of NO(3)(-) ions was 1.66 times higher than that of I(-) ions, and diffusion mobility of SO(4)(2-) ions was half of that for I(-) ions. The theoretical interpretation of experimental counterions exchange rate profiles in terms of the Nernst-Planck equation for interdiffusion also showed higher diffusion mobility of NO(3)(-) ions in the PIM than Cl(-), I(-), and ClO(4)(-) ions, which have comparable diffusion mobility.

Polychloroprene rubber/ethylene-propylene diene monomer/multiple walled carbon nanotube nanocomposites: synergistic effects of radiation crosslinking and MWNT addition

Synergistic effect of MWNT induced reinforcement and high energy radiation induced crosslinking on the physico-mechanical and thermal characteristics of polychloroprene rubber (PCR)/ethylene-propylene diene rubber (EPDM)/MWNT elastomeric nanocomposites was investigated. The extent of reinforcement, as assessed using the Kraus equation suggested high reinforcement of the blend on MWNT addition; though, the thermal stability and glass transition of the PCR and EPDM components were not significantly affected by MWNTs. The elastic modulus increased with the radiation dose as well as with the increase in MWNT content. The reinforcing mechanism of the nano-composites was studied by various micromechanics models all of which predicted higher moduli than the experimentally observed results, indicating agglomeration in the nano-composites. Nevertheless, in all the composites synergistic effect of radiation crosslinking and MWNT induced reinforcement were seen, suggesting radiation induced crosslinking between polymer and MWNT interface.

Counter ion induced irreversible denaturation of hen egg white lysozyme upon electrostatic interaction with iron oxide nanoparticles: A predicted model

The effects of electrostatic interaction between the hen egg white lysozyme (HEWL) and the functionalized iron oxide nanoparticles (IONPs) have been investigated using several techniques, e.g., CD, DSC, ζ-potential, UV-visible spectroscopy, DLS, TEM. Nanoparticles (IONPs) were functionalized with three hydrophilic ligands, viz., poly(ethylene glycol) (PEG), trisodium citrate (TSC) and sodium triphosphate (STP); where both TSC and STP contain Na(+) counter ions. It has been observed that the secondary structure of HEWL was not affected by PEG functionalized IONPs, but was partially and almost completely perturbed by TSC and STP functionalized IONPs, respectively. The perturbation of the secondary structure was irreversible. We have predicted an interaction model to explain the origin of perturbation of HEWL structure. We have also investigated the stability of nanoparticles dispersions after interaction with HEWL and used the DLVO theory to explain results.

NMR Insights into Folding and Self-Association of Plasmodium falciparum P2

The eukaryotic 60S-ribosomal stalk is composed of acidic ribosomal proteins (P1 and P2) and neutral protein P0, which are thought to be associated as a pentameric structure, [2P1, 2P2, P0]. Plasmodium falciparum P2 (PfP2) appears to play additional non-ribosomal functions associated with its tendency for homo-oligomerization. Recombinant bacterially expressed PfP2 protein also undergoes self-association, as shown by SDS-PAGE analysis and light scattering studies. Secondary structure prediction algorithms predict the native PfP2 protein to be largely helical and this is corroborated by circular dichroism investigation. The 1H-15N HSQC spectrum of native P2 showed only 43 cross peaks compared to the expected 138. The observed peaks were found to belong to the C-terminal region, suggesting that this segment is flexible and solvent exposed. In 9 M urea denaturing conditions the chain exhibited mostly non-native β structural propensity. 15N Relaxation data for the denatured state indicated substantial variation in ms-µs time scale motion along the chain. Average area buried upon folding (AABUF) calculations on the monomer enabled identification of hydrophobic patches along the sequence. Interestingly, the segments of slower motion in the denatured state coincided with these hydrophobic patches, suggesting that in the denatured state the monomeric chain undergoes transient hydrophobic collapse. The implications of these results for the folding mechanism and self-association of PfP2 are discussed.

Lecithin-based novel cationic nanocarriers (LeciPlex) I: fabrication, characterization and evaluation.

AIMS In the present investigation, the feasibility of fabricating novel self-assembled cationic nanocarriers (LeciPlex) containing cetyltrimethylammonium bromide (CTAB) or didodecyldimethylammonium bromide (DDAB) and soybean lecithin using pharmaceutically acceptable biocompatible solvents such as 2-Pyrrolidone (Soluphor P) and diethyleneglycol monoethyl ether (Transcutol) was established. MATERIALS & METHODS The interaction between DDAB/CTAB and soybean lecithin in the nanocarriers was confirmed by differential scanning calorimetry and in vitro antimicrobial studies. The positive charge on the nanocarriers was confirmed by zeta potential analysis. RESULTS Transmission electron microscopy analysis could not reveal sufficient information regarding the internal structure of the nanocarriers, whereas cryotransmission electron microscopy studies indicated that these novel nanocarriers have unilamellar structure. Small-angle neutron scattering studies confirmed interaction of cationic surfactant (DDAB) and lecithin in the nanocarriers and confirmed the presence of unilamellar nanostructures. CONCLUSION Various hydrophobic drugs could be encapsulated in the CTAB/DDAB-based lecithin nanocarriers (CTAB-LeciPlex or DDAB-LeciPlex) irrespective of their difference in log p-values. In vitro antimicrobial studies on triclosan-loaded LeciPlex confirmed entrapment of triclosan in the nanocarriers. The ability of CTAB-LeciPlex and DDAB-LeciPlex to condense plasmid DNA was established using agarose gel electrophoresis. DDAB-LeciPlex could successfully transfect pDNA in HEK-293 cells indicating potential in gene delivery.

Interaction of Aspirin with DPPC in the Lyotropic, DPPC-Aspirin-H2O/D2O Membrane

Abstract In recent years, the analgesic, aspirin, has been found to be a platelet aggregation inhibitor. Its interaction with membranes at a molecular level is not yet understood. In this paper, we describe DSC and proton NMR work, carried out to study aspirin-DPPC interaction in the lyotropic, DPPC-water model membrane. Our results show that (1) the aromatic group of aspirin interacts significantly with the glycerol moiety of DPPC and (2) aspirin decreases the chain melting transition point, thus enhancing the permeability of the membrane.

Selective binding of proteins on functional nanoparticles via reverse charge parity model: an in vitro study

The conformation of proteins absorbed on nanoparticles surface plays a crucial role in applications of nanoparticles in biomedicine. The surface protein conformation depends on several factors, namely, nature of protein-nanoparticles interaction, chemical composition of the surface of nanoparticles etc. A model of the electrostatic binding of proteins on charged surface nanoparticles has been proposed earlier (Ghosh et al 2013 Colloids Surf. B 103 267). Also, the irreversible denaturation of the protein conformation due to binding of counterions was reported. In this paper, we have used this model, involving reverse charge parity, to show selective binding of proteins on charged surface iron oxide nanoparticles (IONPs). IONPs were surface functionalized with cetylpyridinium chloride (CPC), cetyl(trimethyl)ammonium bromide (CTAB) and cetylpyridinium iodide (CPI). The effect of counterions (Cl−, Br− and I−) on protein conformation has also been investigated. Several proteins such as α-lactalbumin (ALA), β-lactoglobulin (BLG), ovalbumin (OVA), bovin serum albumin (BSA) and HEWL were chosen for this investigation.

Influence of salicylic acid on the biophysical properties of dipalmitoyl phosphatidylcholine vesicles

The effect of the keratolytic drug salicylic acid (SA) on the thermotropic behaviour, and dynamics of dipalmitoyl phosphatidyl choline (DPPC)–water/buffer pH 7.4 vesicles was studied using DSC and 1H NMR. In both systems, incorporation of SA in DPPC bilayer causes a significant depression in the transition temperature of both the pre-transition (PT) and the gel-to-liquid crystalline (CM) transition. The presence of the drug reduces the cooperativity of both the PT and CM transitions. These findings indicate that SA is bound strongly to the lipid bilayer leading to increased membrane fluidity. The DPPC vesicles incorporated with high drug concentration show phase segregation. One of the interesting findings in this study is the formation of a more ordered high temperature gel (Lβ2) phase when the SA-doped DPPC dispersion is prepared at physiological pH. The effect of inclusion of cholesterol in the SA-free and SA-doped DPPC dispersion was also studied.

Unfolding and inactivation of proteins by counterions in protein-nanoparticles interaction.

In this work, the structure and activity of proteins; such as, hen egg lysozyme (HEWL) and calf intestine alkaline phosphatase (CIAP); have been investigated after incubation with surface coated iron oxide nanoparticles (IONPs) in water. IONPs were coated with counterions bound charge-ligands and were named as the charge-ligand counterions iron oxide nanoparticles (CLC-IONPs). The coating was done with tri-lithium citrate (TLC) and tri-potassium citrate (TKC) to have negative surface charge of CLC-IONPs and Li(+) and K(+), respectively, as counterions. To have positive surface charge, IONPs were coated with cetylpyridinium chloride (CPC) and cetylpyridinium iodide (CPI) having Cl(-) and I(-), respectively, as counterions. The secondary structure of proteins was measured using far ultraviolet circular dichroism (CD) spectroscopy which showed that both proteins were irreversibly unfolded after incubation with CLC-IONPs. The unfolded proteins were seen to be functionally inactive, as confirmed through their activity assays, i.e., HEWL with Escherichia coli (E. coli) and CIAP with para-nitrophenyl phosphate (pNPP). Additionally, we have observed that monomeric hemoglobin (Hb) from radio-resistant insect Chironomus ramosus (ChHb) was also partially unfolded upon interaction with CLC-IONPs. This work clearly shows the role of counterions in protein inactivation via protein-nanoparticles interaction and, therefore, CLC-IONPs could be used for therapeutic purpose.

Interaction of keratolytic drug, salicylic acid with dipalmitoyl phosphatidylethanolamine vesicles

Salicylic acid (SA), a keratolytic drug, is used to treat skin disorder like corns, warts, and acne. To understand the mechanism by which SA interacts with the cell membranes, we have investigated its interaction with dipalmitoyl phosphatidylethanolamine (DPPE) vesicles using DSC, 1H NMR and Raman spectroscopy. Presence of drug asymmetrically broadened the acyl chain melting transition and shifted the transition temperature, T m, to lower value. Both, NMR and DSC studies indicate that the drug molecules are located in the glycerol backbone region of the lipid bilayer and increase the membrane headgroup fluidity. At high drug concentration, additional transitions are observed whose intensity increases with increasing drug concentration. In cholesterol doped DPPE dispersion the interaction of SA with DPPE bilayer is more. The transformation of the gel phase of DPPE dispersion to a stable crystalline subgel phase(s) is accelerated by the presence of SA.