Efficacious redox-responsive gene delivery in serum by ferrocenylated monomeric and dimeric cationic cholesterols

Herein, we present the design and synthesis of new redox-active monomeric and dimeric (gemini) cationic lipids based on ferrocenylated cholesterol derivatives for gene delivery. The cationic cholesterols are shown to be transfection efficient after being formulated with the neutral helper lipid DOPE in the presence of serum (FBS). The redox activity of the resulting co-liposomes and their lipoplexes could be regulated using the alkanyl ferrocene moiety attached to the ammonium head groups of the cationic cholesterols. Atomic force microscopy (AFM), dynamic light scattering (DLS) and zeta potential measurements were performed to characterize the co-liposomal aggregates and their complexes with pDNA. The transfection efficiency of lipoplexes could be tuned by changing the oxidation state of the ferrocene moiety. The gene transfection capability was assayed in terms of green fluorescence protein (GFP) expression using pEGFP-C3 plasmid DNA in three cell lines of different origins, namely Caco-2, HEK293T and HeLa, in the presence of serum. The vesicles possessing ferrocene in the reduced state induced an efficient transfection, even better than a commercial reagent Lipofectamine 2000 (Lipo 2000) as evidenced by flow cytometry and fluorescence microscopy. All the co-liposomes containing the oxidized ferrocene displayed diminished levels of gene expression. Gene transfection events from the oxidized co-liposomes were further potentiated by introducing ascorbic acid (AA) as a reducing agent during lipoplex incubation with cells, leading to the resumption of transfection activity. Assessment of transfection capability of both reduced and oxidized co-liposomes was also undertaken following cellular internalization of labelled pDNA using confocal microscopy and flow cytometry. Overall, we demonstrate here controlled gene transfection activities using redox-driven, transfection efficient cationic monomeric and dimeric cholesterol lipids. Such systems could be used in gene delivery applications where transfection needs to be performed spatially or temporally.

Mapping the intriguing transient morphologies and the demixing behavior in PS/PVME blends in the presence of rod-like nanoparticles

The demixing behavior, transient morphologies and mechanism of phase separation in PS/PVME blends were greatly altered in the presence of a very low concentration of rod-like particles (multiwall carbon nanotubes, MWNTs). This phenomenon is due to the specific interaction of one of the phases (PVME) with the anisotropic MWNTs, which creates a heterogeneous environment in the blend. This specific interaction alters the chain dynamics in the interfacial region as against the bulk. A comprehensive analysis using isochronal temperature sweep was performed to understand the demixing temperature in the blends. The evolution of phase morphology as a function of time and temperature was assessed by polarizing optical microscopy (POM), atomic force microscopy (AFM) and scanning electron microscopy (SEM). The addition of MWNTs increased the rheological demixing temperature and the spinodal temperature in almost all the compositions. The intriguing transient morphologies were mapped, which varied from nucleation and growth to coalescence-induced viscoelastic phase separation (C-VPS) in PVME-rich blends, to spinodal decomposition in the near-critical compositions, to transient gel-induced VPS (T-VPS) in the PS-rich compositions. Mapping of the morphology development displayed two types of fracture mechanisms: ductile fracture for near-critical compositions and brittle fracture for off-critical composition. The change in the phase separation mechanism in the presence of MWNTs was due to the variation in dynamic asymmetry brought about by these anisotropic particles. All these observations were correlated by POM, SEM and AFM studies. The length of the cooperatively rearranging region (CRR), as evaluated using modulated differential scanning calorimetry (MDSC) measurements, was found to be composition-independent. The observed variation of effective glass transition of PVME (low T-g component) on blending with PS (high Tg component) and by the addition of MWNTs accounts for the dynamic heterogeneity introduced by MWNTs in the system.

Instability Of Crack Propagation In Brittle Bulk Metallic Glass

The instability of the crack tip in brittle Mg-based bulk metallic glass (BMG) is studied. The formation of various fractographic surfaces of the BMG is associated with the instability of the fluid meniscus, which is due to viscous fluid matter being present on the fracture process zone. Depending on the values of the wavelength of the initial perturbation of the fluid meniscus and the local stress intensity factor, different fracture surface profiles, i.e. a dimple-like structure, a periodic corrugation pattern and a pure mirror zone are formed. The fractographic evolution is significantly affected by the applied stress. A decreased fracture Surface roughness is observed under a low applied stress. An increased fracture surface roughness, which has frequently been reported by other researchers, is also observed in the present studies under a high applied stress. Unique fractographic features are attributed to the non-linear hyperelastic stiffening for less softening) mechanism. (C) 2008 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Competitive adsorption between bovine serum albumin and collagen observed by atomic force microscope

Atomic force microscopy (AFM) was used to study the competitive adsorption between bovine serum albumin (BSA) and type I collagen on hydrophilic and hydrophobic silicon wafers. BSA showed a grain shape and the type I collagen displayed fibril-like molecules with relatively homogeneous height and width, characterized with clear twisting (helical formation). These AFM images illustrated that quite a lot of type I collagen appeared in the adsorption layer on hydrophilic surface in a competitive adsorption state, but the adsorption of BSA was more preponderant than that of type I collagen on hydrophobic silicon wafer surface. The experiments showed that the influence of BSA on type I collagen adsorption on hydrophilic surface was less than that on hydrophobic surface.

分子间相互作用对蛋白质晶体生长的影响

采用原子力显微镜对溶菌酶和刀豆蛋白A的分子间相互作用力的情况进行了研究,并用动态光散射研究了此二种分子间相互作用力有较大差异的蛋白质在晶体生长条件和非生长条件下,溶液中的聚集体的状态(大小和分散度)随浓度和温度的变化情况.实验结果表明,范德华力强的刀豆蛋白A在成核前,溶液中的聚集体不能很快转变为生长基元,导致晶体生长时间长;而范德华力弱的溶菌酶,溶液中的聚集体可以很快转变成生长基元,晶体生长时间也较短.

Read-only memory disk with AgOx and AgInSbTe superresolution mask layer

Two novel read-only memory (ROM) disks, one with an AgOx mask layer and the other with an AgInSbTe mask layer, are proposed and studied. The AgOx and the AgInSbTe films sputtered on the premastered substrates with pit depths of 50 nm and pit lengths (space) of 380 nm are studied by atomic force microscopy. Disk readout measurement is carried out using a dynamic setup with a laser wavelength of 632.8 nm and an object lens numerical aperture (NA) of 0.40. Results show that the superresolution effect happens only at a suitable oxygen flow ratio for the AgOx ROM disk. The best superresolution readout effect is achieved at an oxygen flow ratio of 0.5 with the smoothest film surface. Compared with the AgOx ROM disk, the AgInSbTe ROM disk has a much smoother film surface and better superresolution effect. A carrier-to-noise ratio (CNR) of above 40 dB can be obtained at an appropriate readout power and readout velocity. The readout CNR of both the AgOx and AgInSbTe ROM disks have a nonlinear dependence on the readout power. The superresolution readout mechanisms for these ROM disks are analyzed and compared as well. (c) 2005 Society of Photo-Optical Instrumentation Engineers.

Read-only memory disk with AgOx super-resolution mask layer

A novel read-only memory (ROM) disk with an AgOx mask layer was proposed and studied in this letter. The AgOx films sputtered on the premastered substrates, with pits depth of 50 nm and pits length of 380 nm, were studied by an atomic force microscopy. The transmittances of these AgOx films were also measured by a spectrophotometer. Disk measurement was carried out by a dynamic setup with a laser wavelength of 632.8 nm and a lens numerical aperture (NA) of 0.40. The readout resolution limit of this setup was λ/(4NA) (400 nm). Results showed that the super-resolution readout happened only when the oxygen flow ratios were at suitable values for these disks. The best super-resolution performance was achieved at the oxygen flow ratio of 0.5 with the smoothest film surface. The super-resolution readout mechanism of these ROM disks was analyzed as well.

Read-only memory disk with AgOx and AgInSbTe superresolution mask layer

Two novel read-only memory (ROM) disks, one with an AgOx mask layer and the other with an AgInSbTe mask layer, are proposed and studied. The AgOx and the AgInSbTe films sputtered on the premastered substrates with pit depths of 50 nm and pit lengths (space) of 380 nm are studied by atomic force microscopy. Disk readout measurement is carried out using a dynamic setup with a laser wavelength of 632.8 nm and an object lens numerical aperture (NA) of 0.40. Results show that the superresolution effect happens only at a suitable oxygen flow ratio for the AgOx ROM disk. The best superresolution readout effect is achieved at an oxygen flow ratio of 0.5 with the smoothest film surface. Compared with the AgOx ROM disk, the AgInSbTe ROM disk has a much smoother film surface and better superresolution effect. A carrier-to-noise ratio (CNR) of above 40 dB can be obtained at an appropriate readout power and readout velocity. The readout CNR of both the AgOx and AgInSbTe ROM disks have a nonlinear dependence on the readout power. The superresolution readout mechanisms for these ROM disks are analyzed and compared as well. (c) 2005 Society of Photo-Optical Instrumentation Engineers.

α-Synuclein senses lipid packing defects and induces lateral expansion of lipids leading to membrane remodeling.

There is increasing evidence for the involvement of lipid membranes in both the functional and pathological properties of α-synuclein (α-Syn). Despite many investigations to characterize the binding of α-Syn to membranes, there is still a lack of understanding of the binding mode linking the properties of lipid membranes to α-Syn insertion into these dynamic structures. Using a combination of an optical biosensing technique and in situ atomic force microscopy, we show that the binding strength of α-Syn is related to the specificity of the lipid environment (the lipid chemistry and steric properties within a bilayer structure) and to the ability of the membranes to accommodate and remodel upon the interaction of α-Syn with lipid membranes. We show that this interaction results in the insertion of α-Syn into the region of the headgroups, inducing a lateral expansion of lipid molecules that can progress to further bilayer remodeling, such as membrane thinning and expansion of lipids out of the membrane plane. We provide new insights into the affinity of α-Syn for lipid packing defects found in vesicles of high curvature and in planar membranes with cone-shaped lipids and suggest a comprehensive model of the interaction between α-Syn and lipid bilayers. The ability of α-Syn to sense lipid packing defects and to remodel membrane structure supports its proposed role in vesicle trafficking.

Evaluation of the functionality of biodegradable polymeric platforms for drug delivery systems

We present the development of a drug-loaded triple-layer platform consisting of thin film biodegradable polymers, in a properly designed form for the desired gradual degradation. Poly(dl-lactide-co-glycolide) (PLGA (65:35), PLGA (75:25)) and polycaprolactone (PCL) were grown by spin coating technique, to synthesize the platforms with the order PCL/PLGA (75:25)/PLGA (65:35) that determine their degradation rates. The outer PLGA (65:35) layer was loaded with dipyridamole, an antiplatelet drug. Spectroscopic ellipsometry (SE) in the Vis-far UV range was used to determine the nanostructure, as well as the content of the incorporated drug in the as-grown platforms. In situ and real-time SE measurements were carried out using a liquid cell for the dynamic evaluation of the fibrinogen and albumin protein adsorption processes. Atomic force microscopy studies justified the SE results concerning the nanopores formation in the polymeric platforms, and the dominant adsorption mechanisms of the proteins, which were defined by the drug incorporation in the platforms. © 2013 Elsevier B.V. All rights reserved.

Dielectric properties of Bi2Ti2O7 films grown on Si(100) substrate by APMOCVD

The growth of Bi2Ti2O7 films with (111) orientation on Si(100) substrate by atmospheric pressure metal-organic chemical vapor deposition(APMOCVD) technique at 480similar to550 degreesC is presented. The films were characterized by X-ray diffraction analysis, atomic force microscopy and electron diffraction. The results show high quality Bi2Ti2O7 films with smooth shinning surface. The dielectric properties and C-V characterization of the films were studied. The dielectric constant (epsilon) and loss tangent (tgdelta) were found to be 180 and 0.01 respectively. The charge storage density was 31.9fC/mum(2). The resistivity is higher than 1x10(12) Omega. .cm under the applied voltage of 5V. The Bi2Ti2O7 films are suitable to be used as a new insulating gate material in dynamic random access memory (DRAM).

Analysis of atomic force microscopic results of InAs islands formed by molecular beam epitaxy

Atomic force microscopy (AFM) measurements of nanometer-sized islands formed by 2 monolayers of InAs by molecular beam epitaxy have been carried out and the scan line of individual islands was extracted from raw AFM data for investigation. It is found that the base widths of nanometer-sized islands obtained by AFM are not reliable due to the finite size and shape of the contacting probe. A simple model is proposed to analyze the deviation of the measured value From the real value of the base width of InAs islands. (C) 1998 Elsevier Science B.V. All rights reserved.

Dielectric properties of Bi2Ti2O7 films grown on Si(100) substrate by APMOCVD

The growth of Bi2Ti2O7 films with (111) orientation on Si(100) substrate by atmospheric pressure metal-organic chemical vapor deposition(APMOCVD) technique at 480similar to550 degreesC is presented. The films were characterized by X-ray diffraction analysis, atomic force microscopy and electron diffraction. The results show high quality Bi2Ti2O7 films with smooth shinning surface. The dielectric properties and C-V characterization of the films were studied. The dielectric constant (epsilon) and loss tangent (tgdelta) were found to be 180 and 0.01 respectively. The charge storage density was 31.9fC/mum(2). The resistivity is higher than 1x10(12) Omega. .cm under the applied voltage of 5V. The Bi2Ti2O7 films are suitable to be used as a new insulating gate material in dynamic random access memory (DRAM).

Fractals in fracture of bulk metallic glass

We investigate the nanoscale periodic corrugation (NPC) structures on the dynamic fracture surface of a typical tough bulk metallic glass, submitted to high-velocity plate impact and scanned by atomic force microscopy (AFM). The detrended fluctuation analysis (DFA) of the recorded AFM profiles reveals that the valley landscapes of the NPC are nearly memoryless, characterized by Hurst parameter of 0.52 and exhibiting a self-similar fractal character with the dimension of about 1.48. Our findings confirm the existence of the “quasi-cleavage” fracture underpinned by tension transformation zones (TTZs) in metallic glasses.

Atomic force microscopic study of low temperature induced disassembly of RecA-dsDNA filaments

The assembly and disassembly of RecA-DNA nucleoprotein filaments on double-stranded DNA (dsDNA) or single-stranded DNA (ssDNA) are important steps for homologous recombination and DNA repair. The assembly and disassembly of the nucleoprotein filaments are sensitive to the reaction conditions. In this work, we investigated different morphologies of the formed nucleoprotein filaments at low temperature under different solution conditions by atomic force microscopy (AFM). We found that low temperature and long keeping time could induce the incomplete disassembly of the formed nucleoprotein filaments.