Hydrophilic Outer Surface Research Articles

Homogeneous polyimide/cyclodextrin composite membranes for pervaporation dehydration of isopropanol

Composite membranes with a sub-nanoscale homogeneous distribution of CD toroids in the Matrimid matrix were developed for dehydration of aqueous isopropanol. The composite membranes demonstrated separation factor far surpassing that of the neat Matrimid dense membrane. The heart of this innovation is the utilization of a CD derivative, ethylenediamine-β-cyclodextrin (EDA-β-CD), where the amine of CD could react with the imide of Matrimid and efficiently immobilize the CD rings during membrane formation. The superior separation properties for membranes embedded with 2–5% EDA-β-CD were attributed to the additional water channels created by the hydrophilic outer surface of CD and its interactions with the polymer matrix. FT-IR, density measurements and XRD have confirmed these hypotheses. Nevertheless, the separation factor exhibited an increasing then decreasing trend as a function of CD content and the opposite trend was observed with permeation flux. Investigation on the effect of feed water concentration showed that the neat Matrimid membrane possessed almost constant performance, but the Matrimid/EDA-β-CD (0.05) composite membrane exhibited an obvious increase in permeability and a decrease in selectivity at high water content. Even though the composite membrane swelled more at higher water content due to the intensified hydrophilicity ascribed to the introduction of CD structure, it always had much better separation factor. In addition, the Matrimid mixed matrix membranes embedded with 2–5% EDA-β-CD held reasonably tensile strength and modulus. The newly developed mixed matrix membrane approach may open up a new way to prepare next-generation high-performance asymmetric pervaporation membranes for isopropanol separation.

Biointerfacing luminescent nanotubes

Supported lipid bilayers that mimic a cell membrane are well-defined model systems for fundamental research and also are critical for the development of new types of biosensor, biodevices and functional materials. We report here that supported lipid bilayers could spontaneously wrap around the layer-by-layer assembled luminescent nanotubes with a hydrophilic polymer-cushioned outer surface. We show that such one-dimensional lipid membranes are laterally mobile and have a high mobile fraction. We also used a simplified diffusion model to estimate the mobility of lipid molecules. These biointerfacing nanotubes have great potential for the design of novel biosensors and biomimetic functional nanomaterials.

Binding of brilliant red compound to lysozyme: insights into the enzyme toxicity of water-soluble aromatic chemicals

The non-covalent interaction of brilliant red (BR) with lysozyme was investigated by the UV spectrometry, circular dichroism (CD) and isothermal titration calorimetry (ITC). The thermodynamic characterization of the interaction was performed and the assembly complexes were formed: lysozyme(BR)(17) at pH 2.03, lysozyme(BR)(15) at pH 3.25 and lysozyme(BR)(12) at pH 4.35, which corresponded to the physiological acidities. The ionic interaction induces a combination of multiple non-covalent bonds including hydrogen bond, hydrophobic interaction and van der Waals force. The two-step binding model of BR was found, in which one or two BR molecules entered the hydrophobic intracavity of lysozyme and the others bound to the hydrophilic outer surface of lysozyme. Moreover, BR binding resulted in change of the lysozyme conformation and inhibition of the lysozyme activity. The possible binding site and type of BR and the conformational transition of lysozyme were speculated and illustrated. This work provided a useful approach for study on enzyme toxicity of aromatic azo chemicals.

Protection by cholesterol‐extracting cyclodextrins: a role for N‐methyl‐d‐aspartate receptor redistribution

Cyclodextrins (CDs) are cyclic oligosaccharides composed of a lipophilic central cavity and a hydrophilic outer surface. Some CDs are capable of extracting cholesterol from cell membranes and can affect function of receptors and proteins localized in cholesterol-rich membrane domains. In this report, we demonstrate the neuroprotective activity of some CD derivatives against oxygen-glucose deprivation (OGD), N-methyl-D-aspartic acid (NMDA) and glutamate in cortical neuronal cultures. Although all CDs complexed with NMDA or glutamate, only beta-, methylated beta- and sulfated beta-CDs displayed neuroprotective activity and lowered cellular cholesterol. Only CDs that lowered cholesterol levels redistributed the NMDA receptor NR2B subunit, PSD-95 (postsynaptic density protein 95 kDa) and neuronal nitric oxide synthase (nNOS) from Triton X-100 insoluble membrane domains to soluble fractions. Cholesterol repletion counteracted the ability of methylated beta-CD to protect against NMDA toxicity, and reversed NR2B, PSD-95 and nNOS localization to Triton X-100 insoluble membrane fraction. Surprisingly, neuroprotective CDs had minimal effect on NMDA receptor-mediated increases in intracellular Ca(2+) concentration ([Ca(2+)](i)), but did suppress OGD-induced increases in [Ca(2+)](i). beta-CD, but not Mbeta-CD, also caused a slight block of NMDA-induced currents, suggesting a minor contribution to neuroprotection by direct action on NMDA receptors. Taken together, data suggest that cholesterol extraction from detergent-resistant microdomains affects NMDA receptor subunit distribution and signal propagation, resulting in neuroprotection of cortical neuronal cultures against ischemic and excitotoxic insults. Since cholesterol-rich membrane domains exist in neuronal postsynaptic densities, these results imply that synaptic NMDA receptor subpopulations underlie excitotoxicity, which can be targeted by CDs without affecting overall neuronal Ca(2+) levels.

Cyclodextrins in eye drop formulations: enhanced topical delivery of corticosteroids to the eye.

Cyclodextrins are cylindrical oligosaccharides with a lipophilic central cavity and hydrophilic outer surface. They can form water-soluble complexes with lipophilic drugs, which 'hide' in the cavity. Cyclodextrins can be used to form aqueous eye drop solutions with lipophilic drugs, such as steroids and some carbonic anhydrase inhibitors. The cyclodextrins increase the water solubility of the drug, enhance drug absorption into the eye, improve aqueous stability and reduce local irritation. Cyclodextrins are useful excipients in eye drop formulations of various drugs, including steroids of any kind, carbonic anhydrase inhibitors, pilocarpine, cyclosporins, etc. Their use in ophthalmology has already begun and is likely to expand the selection of drugs available as eye drops. In this paper we review the properties of cyclodextrins and their application in eye drop formulations, of which their use in the formulation of dexamethasone eye drops is an example. Cyclodextrins have been used to formulate eye drops containing corticosteroids, such as dexamethasone, with levels of concentration and ocular absorption which, according to human and animal studies, are many times those seen with presently available formulations. Cyclodextrin-based dexamethasone eye drops are well tolerated in the eye and seem to provide a higher degree of bioavailability and clinical efficiency than the steroid eye drop formulations presently available. Such formulations offer the possibility of once per day application of corticosteroid eye drops after eye surgery, and more intensive topical steroid treatment in severe inflammation. While cyclodextrins have been known for more than a century, their use in ophthalmology is just starting. Cyclodextrins are useful excipients in eye drop formulations for a variety of lipophilic drugs. They will facilitate eye drop formulations for drugs that otherwise might not be available for topical use, while improving absorption and stability and decreasing local irritation.

Self-Assembly of Dendritic Micellar Structures Based on Triton X-100 and Branched Poly(ethylene oxide)-containing Cyclodextrins

Interaction between the branched poly(etylene oxide)-containing cyclodextrins and nonionic surfactant Triton X-100 was studied. Amphiphilic character of substituted cyclodextrins where poly(etylene oxide) fragments serve as the hydrophilic moiety and outer cyclodextrin surface, as the hydrophobic moiety. Using two groups of experimental methods, it was shown that substituted β-cyclodextrin forms inclusion complexes with hydrophilic poly(ethylene oxide) fragment of Triton X-100 molecule, surfactant molecules being incorporated into a micelle composition. The first group of these methods includes viscometry of solutions and measurement of rotational diffusion coefficient, while the other group (tensiometry, gel permeation chromatography, and spectrophotometry) makes it possible to determine colloidal properties of components. The modified micelles consist of nonionic surfactant molecules, whose outer ends are strung with molecules of substituted cyclodextrins; moreover, their structure resembles that of dendrimers due to arising branchings. Note that the self-assembly method was employed to prepare such dendrimers for the first time.

Electrodialytic transport properties of cation exchange membranes in the presence of cyclodextrins

After commercial cation exchange membranes (NEOSEPTA) had been immersed in an aqueous cyclodextrin solutions, electrodialysis of a 1:1 mixed solutions of alkaline earth metal ions and sodium chloride was carried out in the presence of cyclodextrins in the desalting side solution. Adsorption of the cyclodextrins in the membrane matrix was confirmed by IR spectrum, X-ray photoelectron spectroscopy (XPS) and the weight increase of each corresponding component of the membranes. As a result of adsorption of the cyclodextrins in the membranes, transport numbers of alkaline earth metal ions relative to sodium ions decreased compared with those of the membranes without cyclodextrins. Water content in the membrane increased and a ratio of calcium ions (alkaline earth metal ions) to sodium ions in the membrane phase decreased after adsorption of the cyclodextrins. This is due to existence of the compounds having hydrophilic outer surface, cyclodextrins, in the membrane matrix. The decrease in the transport number of calcium ions relative to sodium ions was due to both decrease in a ratio of mobility of calcium ions to that of sodium ions and decrease in an ionic ratio of calcium ions to sodium ions in the membrane phase.

Unzipping a Membrane

Abstract The atomic force microscope (AFM) is well known for its outstanding spatial resolution, but it is becoming increasingly useful as the instrument for force spectroscopy. In the force spectroscopy mode, the AFM can measure tiny tension forces, in the piconewton (pN) range. Daniel Müller, Wolfgang Baurmeister, and Andreas Engel have used the AFM in both the imaging and force spectroscopy modes to pull proteins out of membranes in a controlled fashion. Müller et al. used Deinococcus radiodurans, a bacterium best known for its high resistance to radiation (as its Genus name implies), as their test subject. They extracted a highly regular membrane from the bacterium, the hexagonally packed intermediate (HPl) layer. They mounted the HPI on mica, so that the hydrophilic outer surface of the HPI adsorbed strongly to the mica, exposing the hydrophobic inner surface to the silicon nitride AFM stylus.

An air-stable biomimetic Langmuir–Blodgett bilayer

The long-chain amphiphiles in biomembranes have their hydrophilic headgroups oriented outwards towards the aqueous environment, and their long aliphatic chains innermost. In Langmuir–Blodgett (LB) films the hydrophobic chains are normally outermost. LB films of long-chain derivatives can be prepared under water with the headgroups outermost and are stable while kept there, but the act of removing them from the aqueous environment removes the top monolayer. We demonstrate an adaptation of the LB technique which is appropriate for the preparation of biocompatible surfaces. It produces essentially bilayer films of long-chain derivatives on standard substrates with a hydrophilic outer surface. The films are stable in air. The films show large contact angle hysteresis with water, but the receding angle is in most cases less than 10°. The films can be immersed and withdrawn from water many times before wettability or average thickness change significantly. Compared to previously reported air-stable surfaces with a structure mimicking biological membranes, the substrate for the present surface is experimentally more convenient.

Pure-Silica Zeolites (Porosils) as Model Solids for the Evaluation of the Physicochemical Features Determining Silica Toxicity to Macrophages

The interaction between inhaled particles and alveolar macrophages plays a key role in silica-related diseases. It has been previously shown [Fubini, B., et al. (1999) Chem. Res. Toxicol. 12, 737-745] that a monocyte-macrophage cell line (J774) may be employed in the evaluation of the degree of cytotoxicity to alveolar macrophages of various silica dusts. In this paper, pure-silica zeolites (porosils) in microcrystalline form have been employed as "model solids" in an effort to show which physicochemical properties of the silica particle are playing a major role in the toxicity to macrophages. The samples employed covered four different porosil crystal structures (MFI, FAU, TON, and MTT) and also include a synthetic rodlike cristobalite (CRIS-rd). When compared at equal weight, the samples cover a wide range of cytotoxicity from inert to toxic as unheated mineral cristobalite [Fubini, B., et al. (1999) Chem. Res. Toxicol. 12, 737-745]. Mild grinding did not affect cytotoxicity. Calcined (open pores) and uncalcined (pore filled with template) TON exhibited the same cytotoxicity, indicating that only the outer surface is implied. The hydrophobic and/or hydrophilic character of TON, evaluated by adsorption calorimetry, is close to what has been previously found for silicalite and is consistent with a hydrophilic outer surface and hydrophobic pore walls. The potential for generating hydroxyl radicals from hydrogen peroxide varies among the various porosils that have been studied. A model is proposed for the correlation between inhibition of growth on proliferating cells and physicochemical properties varying from one to the other sample. The extent of external surface and the aspect ratio were related to the intensity of the cytotoxic effect, while the level of radical release was not. This suggests, on one hand, that comparison of toxicity among various dusts should be made at equal particle surface and, on the other, that in the model studied, free radical release does not play a crucial role in the primary event of toxicity to alveolar macrophages.

Conformational change of the hexagonally packed intermediate layer of Deinococcus radiodurans monitored by atomic force microscopy

Both surfaces of the hexagonally packed intermediate (HPI) layer of Deinococcus radiodurans were imaged in buffer solution by atomic force microscopy. When adsorbed to freshly cleaved mica, the hydrophilic outer surface of the HPI layer was attached to the substrate and the hydrophobic inner surface was exposed to the stylus. The height of a single HPI layer was 7.0 nm, while overlapping edges of adjacent single layers adsorbed to mica had a height of 14.7 nm. However, double-layered stacks with inner surfaces facing each other exhibited a height of 17.4 nm. These stacks exposed the outer surface to the stylus. The different heights of overlapping layers and stacks are attributed to differences in the interaction between inner and outer surfaces. At high resolution, the inner surface revealed a protruding core with a central pore connected by six emanating arms. The pores exhibited two conformations, one with and the other without a central plug. Individual pores were observed to switch from one state to the other.

Uniform-size hydrophobic polymer-based separation media selectively modified with a hydrophilic external polymeric layer

A simple procedure for the preparation of macroporous hydrophobic styrene-divinylbenzene polymeric separation media with a hydrophilic outer surface has been developed. A hydrophilic monomer and water-soluble polymerization initiator are added to the reaction mixture during the final polymerization step of the preparation of size-monodisperse particles. Because the hydrophobic styrene-divinylbenzene framework of the beads is already formed, and the hydrophilic monomer does not penetrate the pores of the beads that are filled with a hydrophobic porogen, the hydrophilic layer is formed only at the surface of the beads. The hydrophilic monomers used included glycerol monomethacrylate and glycerol dimethacrylate and toluene was used as the porogen for the poly(styrene-co-divinylbenzene) beads. Comparative experiments involving beads with or without a hydrophilic medium showed that the separation selectivity of the media towards hydrophobic solutes remains unchanged. However, the modified medium with a hydrophilic layer could be used to analyse mixtures that also contained large peptide molecules as these do not adsorb at its surface as is the case with the unmodified hydrophobic beads.

Ureteric retrieval net: comparison with stone extraction by Dormia baskets in an in vitro porcine model

To develop a ureteric retrieval net with advantages over existing stone retrieval methods. In vitro experimental trials comparing the ureteric retrieval net with Dormia baskets were performed with porcine ureters and artificial calculi placed in a proximal or distal position. For removing a single, large stone (9 x 6 x 5 mm) the net was slightly better than baskets (100% versus 90%) and it was significantly (P < 0.001) more effective for removing multiple small stones (2 x 2 x 3 mm to 5 x 4 x 3 mm) in one pass (76% versus 40% with three distal stones; 73% versus 13% with three proximal stones; and 53% versus 3% with six proximal stones). This newly developed ureteric retrieval net has a compact design, resulting in minimal resistance during retraction. Its smooth hydrophilic outer surface enhances scooping properties against the luminal wall. The mesh is similar to that in nylon stockings--porous enough to allow flow, yet small enough to capture fragments. It can capture numerous small fragments with one scoop, but can easily disengage them if extraction is not possible. The loop encompasses the entire lumen with the 'pusher' offset to a side wall, allowing maximal space and visualization, and the device can dilate the ureter to facilitate extraction. Clinical studies are in progress to evaluate the full potential of this new device.

Function and biosynthesis of gas vesicles in halophilic Archaea.

The proteinaceous gas vesicles produced by various microorganisms including halophilic Archaea are hollow, gas-filled structures with a hydrophobic inner and a hydrophilic outer surface. The structural components of gas vesicles and their biosynthesis are still under investigation; an 8-kDa polypeptide appears to be the major constituent of the gas-vesicle envelope. Genetic analysis of the halobacterial gas-vesicle synthesis revealed an unexpected complexity: about 14 genes organized in three transcription units are involved in gas-vesicle structure, assembly, and gene regulation. Here we describe the comparison of three different genomic regions encoding gas vesicles in Halobacterium salinarium (p-vac and c-vac regions) and Haloferax mediterranei (mc-vac region) and speculate on the function of the gene products involved in gas-vesicle synthesis.

The protein encoded by gvpC is a minor component of gas vesicles isolated from the cyanobacteria Anabaena flos‐aquae and Microcyctis sp.

The proteins present in gas vesicles of the cyanobacteria Anabaena flos-aquae and Microcystis sp. were separated by SDS-polyacrylamide gel electrophoresis. Each contained a protein of Mr 22K whose N-terminal amino acid sequences showed homology with that of the Calothrix sp. PCC 7601 gvpC gene product. The gvpC gene from A. flos-aquae was cloned and sequenced. The derived amino acid sequence for the gene product indicated a protein, GVPc, of 193 residues and Mr 21985 containing five highly conserved 33 amino acid repeats. The sequence was identical at the N-terminus to that of the Mr 22K protein present in gas vesicles and showed correspondence to seven tryptic peptides isolated from gas vesicles. This establishes that GVPc forms a second protein component of the gas vesicle, in addition to the main constituent, the 70 residue GVPa. Quantitative amino acid analysis of entire gas vesicles reveals that GVPc accounts for only 2.9% of the protein molecules and 8.2% of the mass present: this is insufficient to form the conical end caps of the gas vesicles. It is suggested that GVPc provides the hydrophilic outer surface of the gas vesicle wall; the 33 amino acid repeats may interact with the periodic structure provided by GVPa.

The double helix coiled coil structure of murein lipoprotein from Escherichia coli

A rod-like structure is proposed for the murein lipoprotein of Escherichia coli, built of two parallel unbroken α-helices arranged in a coiled coil of the same type as in the muscle protein tropomyosin. The amino acid sequence has the required regular pattern of hydrophobic amino acids at intervals of three and four residues and the secondary structure predicted from the sequence is 80% helical. A space-filling model confirms that the coiled coil model is stereochemically reasonable, and energy calculations for a series of coils with different radii suggest that the best structure is one with the helix axes 8.25 Å apart. Energyrefined atomic co-ordinates have been calculated which show that the hydrophobic side-chains form a series of close-packed unstrained contacts between the two helices along the entire length of the sequence. On the basis of this study the hexagonal membrane pore model and the segmented helix model proposed by others seem unlikely. The coiled coil has a strongly hydrophilic outer surface, suggesting that the protein has a watery environment within the E. coli cell envelope and is not strictly a membrane protein. Probably only the fatty acid portion of the lipoprotein penetrates into the lipid region of the outer membrane, so that the protein may act as a tie or a spacer between the lipid and the murein wall.

A three-dimensional molecular assembly model of a lipoprotein from the Escherichia coli outer membrane.

From the complete amino-acid sequence of a lipoprotein from the outer membrane of E. coli, a three-dimensional molecular assembly model was constructed. It is proposed that the model provides a tubular hydrophilic channel through the outer membrane, which serves as a passive diffusion pore. An alpha-helix is constructed from the sequence, and six of them are arranged to form a superhelix with a hydrophilic interior and hydrophobic outer surface. The superhelical assembly is stabilized by seven ionic interactions between adjacent alpha-helices. Since the height of the assembly is 76 A, it could be inserted into the outer membrane and span the full 75-A thick membrane. The assembly is stabilized in the outer membrane not only by hydrophobic interaction between the surface of the assembly and the lipid bilayer, but also by three hydrocarbon chains of fatty acids linked to the amino-terminal end of the lipoprotein, which are flipped back along the assembly and inserted into the lipid bilayer of the outer membrane. Any two alpha-helices in an assembly are linked to the peptidoglycan at their carboxyl-terminal ends so that the outer membrane is anchored on the peptidoglycan layer. Six or more alpha-helices can form an assembly of this type. However, assuming that an assembly consists of six helices, there are 1.25 x 10(5) per cell hydrophilic channels of a diameter of 12.5 A and 35% of the cell surface is occupied by the assemblies.