Droplet‐Shooting and Size‐Filtration (DSSF) Method for Synthesis of Cell‐Sized Liposomes with Controlled Lipid Compositions

We report a centrifugal microfluidic method, droplet‐shooting and size‐filtration (DSSF), for the production of cell‐sized liposomes with controlled lipid compositions. This involves the generation of large and small droplets from the tip of a glass capillary and the selective transfer of small droplets through an oil‐water interface, thus resulting in the generation of cell‐sized liposomes. We demonstrate control of the microdomain formation as well as the formation of asymmetric lipid bilayer liposomes of uniform size by the control of lipid composition. The DSSF method involves simple microfluidics and is easy to use. In addition, only a small volume (0.5–2 μL) of sample solution is required for the formation of hundreds of cell‐sized liposomes. We believe that this method can be applied to generate cell‐sized liposomes for a wide variety of uses, such as the construction of artificial cell‐like systems.     

Composite Membrane Formation by Combination of Reaction‐Induced and Nonsolvent‐Induced Phase Separation

A novel method of preparing skinned asymmetric membranes with two distinctive layers is described: a top layer composed of chemically cross‐linked polymer chains (dense layer) and a bottom layer of non‐cross‐linked polymer chains (porous substructure). The method consists of two simple steps that are compatible with industrial membrane fabrication facilities. Unlike conventional processes to prepare asymmetric membranes, with this approach it is possible to finely control the structure and functionalities of the final membrane. The thickness of the dense layer can be easily controlled over several orders of magnitude and targeted functional groups can be readily incorporated in it.

     

Reversible Photochromic Nanofiber Membrane Containing Comb‐Like Poly(octadecyl acrylate) Nanoparticles Used for Ultraviolet Intensity Indicator

A novel reversible photo‐responsive polyvinyl alcohol (PVA)/polyethylenimine (PEI) electrospinning nanofiber membrane (NFM), assembled with photochromic nanoparticles containing spiropyran, is herein presented and the water resistance, mechanical properties as well as photochromic properties are investigated systematically. Here, glutaraldehyde, which can undergo a cross‐linking reaction with PVA/PEI, is selected as a cross‐linker to enhance water resistance and mechanical behavior of the NFM. Photochromic polymer nanoparticles are prepared using octadecyl acrylate as polymer matrix monomer through miniemulsion polymerization. Effect factors of the whole preparation process such as the PVA/PEI mass ratio, the amount of emulsifier, and the amount of photochromic nanoparticles added are regulated to obtain the optimal experimental parameters. The surface morphology and microstructure, water resistance, and mechanical properties of NFMs are studied by thermal field emission scanning electron microscopy, static water contact angle tester, and tensile universal testing machine, respectively. Furthermore, UV–vis spectrophotometer is employed to investigate the photochromic performance and fatigue resistance of NFMs. Overall, analyses reveal that the reversible photo‐responsive electrospinning NFM exhibit excellent photoresponsivity, photoreversibility, and fatigue resistance upon UV irradiation, thus showing promising application in the field of ultraviolet intensity indicator.     

On the Role of Curved Membrane Nanodomains and Passive and Active Skeleton Forces in the Determination of Cell Shape and Membrane Budding

Biological membranes are composed of isotropic and anisotropic curved nanodomains. Anisotropic membrane components, such as Bin/Amphiphysin/Rvs (BAR) superfamily protein domains, could trigger/facilitate the growth of membrane tubular protrusions, while isotropic curved nanodomains may induce undulated (necklace-like) membrane protrusions. We review the role of isotropic and anisotropic membrane nanodomains in stability of tubular and undulated membrane structures generated or stabilized by cyto- or membrane-skeleton. We also describe the theory of spontaneous self-assembly of isotropic curved membrane nanodomains and derive the critical concentration above which the spontaneous necklace-like membrane protrusion growth is favorable. We show that the actin cytoskeleton growth inside the vesicle or cell can change its equilibrium shape, induce higher degree of segregation of membrane nanodomains or even alter the average orientation angle of anisotropic nanodomains such as BAR domains. These effects may indicate whether the actin cytoskeleton role is only to stabilize membrane protrusions or to generate them by stretching the vesicle membrane. Furthermore, we demonstrate that by taking into account the in-plane orientational ordering of anisotropic membrane nanodomains, direct interactions between them and the extrinsic (deviatoric) curvature elasticity, it is possible to explain the experimentally observed stability of oblate (discocyte) shapes of red blood cells in a broad interval of cell reduced volume. Finally, we present results of numerical calculations and Monte-Carlo simulations which indicate that the active forces of membrane skeleton and cytoskeleton applied to plasma membrane may considerably influence cell shape and membrane budding.     

Reversible immobilization of glycoamylase by a variety of Cu2+‐chelated membranes

Glycoamylase (AMG) is an γ‐amylase enzyme which catalyzes the breakdown of large α(1,4)‐linked malto‐oligosaccharides to glucose. It is an extracellular enzyme and is excreted to the culture medium. In this study, AMG was immobilized on a variety of metal affinity membranes, which were prepared by chelating Cu²⁺ ions onto poly(hydroxyethyl methacrylate) (PHEMA) using N‐methacryloyl‐(L )‐histidine methyl ester (MAH), N‐methacryloyl‐(L )‐cysteine methyl ester (MAC), and N‐methacryloyl‐(L )‐phenylalanine methyl ester (MAPA) as metal‐chelating comonomers for reversible immobilization of AMG. The PHEMAH, PHEMAC, PHEMAPA membranes were synthesized by UV‐initiated photo‐polymerization and Cu²⁺ ions were chelated on the membrane surfaces. Cu²⁺‐chelated membranes were characterized by swelling tests, SEM, contact angle measurements, elemental analysis, and FTIR. AMG immobilization on the Cu²⁺‐chelated membranes was performed by using aqueous solutions of different amounts of AMG at different pH values and Cu²⁺ loadings. Durability tests concerning desorption of AMG and reusability of the Cu²⁺‐chelated membranes yielded acceptable results. It was computationally determined that AMG possesses four likely Cu²⁺/Zn²⁺ binding sites, away from the catalytic site, to which the metal‐chelated membranes can be efficiently used. © 2012 Wiley Periodicals, Inc. J Appl Polym Sci, 2012     

Synthesis and Characterization of a New Bifunctionalized,Fluorescent, and Amphiphilic Molecule for Recruiting SH‐Containing Molecules to Membranes

This study describes the synthesis and characterization of an amphiphilic construct intended to recruit SH‐containing molecules to membranes. The construct consists of 1) an aliphatic chain to enable anchoring within membranes, 2) a maleimide moiety to react with the sulfhydryl group of a soluble (bio)molecule, and 3) a fluorescence moiety to allow the construct to be followed by fluorescence spectroscopy and microscopy. It is shown that the construct can be incorporated into preformed membranes, thus allowing application of the approach with biological membranes. The close proximity between the fluorophore and the maleimide moiety within the construct causes fluorescence quenching. This allows monitoring of the reaction with SH‐containing molecules by measurement of increases in fluorescence intensity and lifetime. Notably, the construct distributes into laterally ordered membrane domains of lipid vesicles, which is probably triggered by the length of its membrane anchor. The advantages of the new construct can be employed for several biological, biotechnological, and medicinal applications.     

Synthesis of Functionalized Hydrazines: Facile Homogeneous (N‐Heterocyclic Carbene)‐Palladium(0)‐Catalyzed Diboration and Silaboration of Azobenzenes

The bis(N‐heterocyclic carbene)(diphenylacetylene)palladium complex [Pd(ITMe)₂(PhC≡CPh)] (ITMe=1,3,4,5‐tetramethylimidazol‐2‐ylidene) acts as a highly active pre‐catalyst in the diboration and silaboration of azobenzenes to synthesize a series of novel functionalized hydrazines. The reactions proceed using commercially available diboranes and silaboranes under mild reaction conditions.

     

Predicting Separation of Lower Hydrocarbon from Natural Gas by a Nano‐Porous Membrane using Capillary Condensation

In the present work, the potential of a nano‐porous membrane for predicting the separation of lower hydrocarbons from natural gas by capillary condensation was explored. While a gas permeates through a capillary at a suitable pressure, the adsorbed layer may attain a thickness enough to fill the entire membrane pore. Poiseuille flow of the condensed phase follows. Our computed results have established that for a passage through a nano‐porous membrane, gas having lower condensation pressure condenses in the pores at a pressure which is about an order of magnitude lower than its vapor pressure at the concerned temperature. In the case of propane/methane and butane/methane binary mixtures, propane and butane are preferentially condensed and permeation rates up to 700 g mol/m² s bar for propane and 600 g mol/m² s bar for butane have been achieved at a temperature lower than the critical temperature of the permeating species and higher than the critical temperature of the non‐permeating species. Since methane has a much lower critical temperature than both propane and butane, it gets physically dissolved in the condensed phase of propane, butane in the case of propane/methane and butane/methane binary mixtures, respectively. An equation of state (EOS) approach has been adopted to calculate the fugacity of methane in the gas, as well as in the condensed phase, in order to estimate its solubility. The Peng‐Robinson equation of state was used. Computation of the separation factor for methane/propane and methane/butane was performed over a wide range of temperature, pressure, and gas composition. The separation factor which is expectedly a function of these variables ranged from 0.3–75 for methane/propane and 0.7–140 for methane/butane binary mixtures. It has been established that an acceptable degree of separation is achievable at moderate pressure and at low temperature for the removal of propane and butane from natural gas. The results have the potential to be used for further refinement and optimization of the process conditions so that this strategy can be exploited for large‐scale removal of lower hydrocarbon from natural gas at a low cost.     

Morphological Change of Cell Membrane by Interaction with Domain‐Swapped Cytochrome c Oligomers

Monomeric cyt c has been reported to bind to the mitochondrial membrane by electrostatic and hydrophobic interactions with anionic phospholipids. We have previously shown that domain‐swapped oligomeric cyt c retains the secondary structure of the monomer, and its surface possesses a larger area and more charges compared to the monomer. However, the effect of oligomerization of cyt c on cells has yet to be revealed. Herein, we investigated the interaction of oligomeric cyt c with anionic phospholipid‐containing vesicles and the outer membrane of HeLa cells. Oligomeric cyt c interacted more strongly than monomeric cyt c with anionic phospholipid‐containing vesicles and the outer membrane of HeLa cells. Oligomeric cyt c induced lateral phase separation of lipids in LUVs and GUVs, thereby leading to membrane disruption, whereas monomeric cyt c did not. Morphological changes in HeLa cells resulted from interaction with oligomeric cyt c, but little from interaction with the monomer. These results show that domain‐swapped oligomeric proteins might exhibit properties different to those of monomer in cell systems.     

Mechanically Robust Nanofibrous Xerogel Membrane for One‐Pass Removal of Trace Water in Oil

Removal of trace water from oil is essential for a high‐performance fuel supply system. Trace water in fuel is a crucial contributor to frequent system maintenance and failure. In this work, an electrospun polyacrylic acid/polyvinyl alcohol (PAA/PVA) nanofibrous xerogel membrane (NFM) is prepared to remove trace water in oil. Based on the optimization of the weight ratio and the crosslinking conditions of PAA/PVA, the resultant NFM shows remarkable water‐retaining capacity (swelling ratio of 124.8 g/g) and rapid absorption property. The optimal membrane displays a small average pore size of 523.2 nm, high porosity of 79.6% as well as robust tensile strength of 8.1 MPa. Additionally, after just one pass through a single layer membrane (thickness of 35 ± 3 µm and areal density of 3.5 ± 0.5 g m^?2), the milky oil containing 1 wt% water is clarified and the water content reduced to 130 ppm. With extensive water being absorbed into the 3D network (different with the conventional separation mechanism) as well as good compatibility with oil, a multilayer pleated scheme may provide a practical methodology for enhancing fuel oil properties and performances.     

Ruthenium Nanoparticle‐Catalyzed,Controlled and Chemoselective Hydrogenation of Nitroarenes using Ethanol as a Hydrogen Source

This communication describes a ruthenium nanoparticle‐catalyzed reduction of nitroarenes giving azoxyarenes, azoarenes, or anilines in good to excellent yields using ethanol as a hydrogen source.     

Amphiphilic Counterion Activators for DNA: Stimuli‐Responsive Cation Transporters and Biosensors in Bulk and Lipid Bilayer Membranes

We report that amphiphilic counterions can enable DNA to act as cation carrier, enzyme detector and biosensor. Calf thymus DNA is used as example throughout the study. Evaluation of a series of counterion activators suggests that strong amphiphilicity, alkyl or calix[4]arene tails and guanidinium cations give best results, whereas weak amphiphilicity, bola‐amphiphilicity, planar aryl tails and ammonium cations are less satisfactory for various reasons. In the U‐tube, DNA–counterion complexes can carry cations such as safranin O or p‐xylene‐bis‐pyridinium bromide (DPX) across bulk chloroform membranes, whereas anions such as carboxyfluorescein (CF) and (8‐Hydroxy‐1,3,6‐pyrenetrisulfonate (HPTS) are not transported. Uptake of DNA–counterion complexes into intact vesicles is demonstrated by DNA trapping experiments with internal polylysine. Comparison of results from different assays suggests that DNA–counterion complexes act as cation carriers under mild conditions, whereas pore formation and lysis dominate at higher concentrations. Applicability of DNA–counterion transporters for the detection of enzyme activity is demonstrated with phytate as an inactivating substrate and phytase as a reactivating enzyme. Compatibility with biosensing is exemplified with the fluorometric monitoring of phytate levels in almond extracts. The conceptual significance of these findings is briefly discussed, as are promising perspectives such as the application of DNA chemistry to multianalyte sensing in fluorogenic vesicles.     

Role of RNA Motifs in RNA Interaction with Membrane Lipid Rafts: Implications for Therapeutic Applications of Exosomal RNAs

RNA motifs may promote interactions with exosomes (EXO-motifs) and lipid rafts (RAFT-motifs) that are enriched in exosomal membranes. These interactions can promote selective RNA loading into exosomes. We quantified the affinity between RNA aptamers containing various EXO- and RAFT-motifs and membrane lipid rafts in a liposome model of exosomes by determining the dissociation constants. Analysis of the secondary structure of RNA molecules provided data about the possible location of EXO- and RAFT-motifs within the RNA structure. The affinity of RNAs containing RAFT-motifs (UUGU, UCCC, CUCC, CCCU) and some EXO-motifs (CCCU, UCCU) to rafted liposomes is higher in comparison to aptamers without these motifs, suggesting direct RNA-exosome interaction. We have confirmed these results through the determination of the dissociation constant values of exosome-RNA aptamer complexes. RNAs containing EXO-motifs GGAG or UGAG have substantially lower affinity to lipid rafts, suggesting indirect RNA-exosome interaction via RNA binding proteins. Bioinformatics analysis revealed RNA aptamers containing both raft- and miRNA-binding motifs and involvement of raft-binding motifs UCCCU and CUCCC. A strategy is proposed for using functional RNA aptamers (fRNAa) containing both RAFT-motif and a therapeutic motif (e.g., miRNA inhibitor) to selectively introduce RNAs into exosomes for fRNAa delivery to target cells for personalized therapy.     

Counterion‐Switched Reversibly Hydrophilic and Hydrophobic TiO2‐Incorporated Layer‐By‐Layer Self‐Assembled Membrane for Nanofiltration

The manipulation of surface wettability has been regarded as an efficient strategy to improve the membrane performances. Herein, the counterion‐switched reversibly hydrophilic and hydrophobic surface of TiO₂‐loaded polyelectrolyte membrane are prepared by layer‐by‐layer assembly of poly(sodium 4‐styrene sulfonate) (PSS) and poly(diallydimethyl‐ammoniumchloride (PDDA) containing TiO₂@PDDA nanoparticles (NPs) on the hydrolyzed polyacrylonitrile (PAN) substrate membrane. The obtained polyelectrolyte multilayer (PEM) membranes [PEM‐TiO₂]_4.5⁺X^? (X^? = Cl^?, PFO^? [perfluorooctanoate] etc.) show different hydrophilicity and hydrophobicity with various counterions. The integration of TiO₂ NPs obviously improves the wettability and nanofiltration (NF) performance of PEM membrane for (non)aqueous system of dyes (crystal violet, eriochrome black T) with a high recyclability. The highly hydrophilic [PEM‐TiO₂]_4.5⁺Cl^? (water contact angle [WCA]: 13.2 ± 1.8°) and hydrophobic [PEM‐TiO₂]_4.5⁺PFO^? (WCA: 115.4 ± 2.3°) can be reversibly switched via counterion exchange between Cl^? and PFO^?, verifying the surface with a reversible hydrophilic–hydrophobic transformation. For such membranes, the morphology, wettability, and NF performance rely on the loading of TiO₂@PDDA NPs and surface counterion. Meanwhile, the motion and interaction of water or ethanol in the hydrophilic or hydrophobic membrane are revealed by low‐field nuclear magnetic resonance. This work provides a facile and rapid approach to fabricate smart and tunable wetting surface for potential utilization in (non)aqueous NF separation.     

The Antimicrobial Activity of Sub3 is Dependent on Membrane Binding and Cell‐Penetrating Ability

Because of their high activity against microorganisms and low cytotoxicity, cationic antimicrobial peptides (AMPs) have been explored as the next generation of antibiotics. Although they have common structural features, the modes of action of AMPs are extensively debated, and a single mechanism does not explain the activity of all AMPs reported so far. Here we investigated the mechanism of action of Sub3, an AMP previously designed and optimised from high‐throughput screening with bactenecin as the template. Sub3 has potent activity against Gram‐negative and Gram‐positive bacteria as well as against fungi, but its mechanism of action has remained elusive. By using AFM imaging, ζ potential, flow cytometry and fluorescence methodologies with model membranes and bacterial cells, we found that, although the mechanism of action involves membrane targeting, Sub3 internalises inside bacteria at lethal concentrations without permeabilising the membrane, thus suggesting that its antimicrobial activity might involve both the membrane and intracellular targets. In addition, we found that Sub3 can be internalised into human cells without being toxic. As some bacteria are able to survive intracellularly and consequently evade host defences and antibiotic treatment, our findings suggest that Sub3 could be useful as an intracellular antimicrobial agent for infections that are notoriously difficult to treat.     

Preparation of poly(4‐methyl‐1‐pentene) asymmetric or microporous hollow‐fiber membranes by melt‐spun and cold‐stretch method

Poly(4‐methyl‐1‐pentene) (PMP) hollow fibers were prepared and fabricated into gas separation or microporous membranes by the melt‐spun and cold‐stretched method. PMP resin was melt‐extruded into hollow fibers with cold air as the cooling medium. The effects of take‐up speed and thermotreatment on the mechanical behavior and morphology of the fibers were investigated. Scanning electronic microscope (SEM) photos were used to reveal the geometric structure of the section and surface of the hollow fibers. It was found that the original fiber had an asymmetric structure. A “sandwich” mode was used to describe the formation of this special fine structure. And a series of PMP hollow‐fiber membranes were prepared by subsequent drawing, and it was found that there was a “skin–core” structure on the cross section of these hollow‐fiber membranes. Asymmetric or microporous PMP hollow‐fiber membranes could be obtained by controlling posttreatment conditions. The morphology of these membranes were characterized by SEM, and the gas (oxygen, nitrogen, and carbon dioxide) permeation properties of the membranes was measured. The results indicate that the annealing time of the original fiber and the stretching ratio were the key factors influencing the structure of the resulting membrane. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 100: 2131–2141, 2006