Challenges in theoretical investigations of configurations of lipid membranes

We report some key results in the theoretical investigations of configurations of lipid membranes and present several challenges in this field, which involve (i) the exact solutions to the shape equation of lipid vesicles, (ii) the exact solutions to the governing equations of open lipid membranes, (iii) the neck condition of two-phase vesicles in the budding state, (iv) the nonlocal theory of membrane elasticity, and (v) the relationship between the symmetry and the magnitude of the free energy.     

Nonrotating cosmic strings interacting with gravitational waves in Einstein-Maxwell-dilaton gravity

We present methods for the construction of exact diagonal cylindrically symmetric solutions in a four dimensional low energy limit of string theory, the Einstein-Maxwell-dilaton gravity. The methods allow us to generate exact string backgrounds from known solutions to the equations of Einstein or Einstein gravity coupled to a massless scalar field. We also give and analyze explicit examples of such solutions. It is shown that they are free of curvature singularities,(quasi)regular on the axis of symmetry, asymptotically flat and describe nonrotating cosmic strings interacting with gravitational, dilaton and electromagnetic waves.     

Limited slowdown of endocrine-disruptor diffusion in confined fluid lipid membranes

Self-diffusion rates of lipids and trapped bisphenol A (BPA) are determined in various sizes of confined but fluid membranes by high-field-gradient NMR at 600 MHz. Micelles and vesicles of 3- to 400-nm diameters are used as model membranes to get an insight into the molecular diffusion in such soft environments. The slowdown of BPA and lipid motions is leveled off in 100- and 400-nm vesicles, although the hydrodynamic continuum model gives the aggregate motion slowed inversely to the aggregate size. Instead, the limited motion is related to the intra-membrane fluidity.     

Equilibrium Configurations of Lipid Bilayer Membranes and Carbon Nanostructures

The present article concerns the continuum modelling of the mechanical behaviour and equilibrium shapes of two types of nano-scale objects: fluid lipid bilayer membranes and carbon nanostructures. A unified continuum model is used to handle four different case studies. Two of them consist in representing in analytic form cylindrical and axisymmetric equilibrium configurations of single-wall carbon nanotubes and fluid lipid bilayer membranes subjected to uniform hydrostatic pressure. The third one is concerned with determination of possible shapes of junctions between a single-wall carbon nanotube and a flat graphene sheet or another single-wall carbon nanotube. The last one deals with the mechanical behaviour of closed fluid lipid bilayer membranes (vesicles) adhering onto a flat homogeneous rigid substrate subjected to micro-injection and uniform hydrostatic pressure.     

Miscibility phase diagrams of giant vesicles containing sphingomyelin

Saturated sphingomyelin (SM) lipids are implicated in lipid rafts in cell plasma membranes. Here we use fluorescence microscopy to observe coexisting liquid domains in vesicles containing SM, an unsaturated phosphatidylcholine lipid (either DOPC or POPC), and cholesterol. We note similar phase behavior in a model membrane mixture without SM (DOPC/DPPC/Chol), but find no micron-scale liquid domains in membranes of POPC/PSM/Chol. We delineate the onset of solid phases below the miscibility transition temperature, and detail indirect evidence for a three-phase coexistence of one solid and two liquid phases.     

多聚赖氨酸诱导的负电性磷脂巨囊泡形变

利用荧光显微技术表征了多聚赖氨酸诱导的负电性磷脂巨囊泡的动力学响应行为.研究发现,多聚赖氨酸可吸附至二油酰磷脂酰胆碱和二油酰磷脂酸混合磷脂巨囊泡的表面,诱导其发生粘连、出"绳"及破裂现象.分析认为,在低盐环境中,膜形变由多聚赖氨酸吸附于二油酰磷脂酸富集区引起的膜两叶应力不对称,以及静电相互作用等因素产生.研究结果对基于聚合物-巨囊泡体系的药物输运控释、细胞形变、微控反应和基因治疗等方面的研究提供有价值的支持.     

A Novel Squarylium Dye for Monitoring Oxidative Processes in Lipid Membranes

A novel squaraine probe SQ-1 has been found to be appropriate for monitoring the peroxidation processes in membrane systems. Formation of free radicals was triggered by methemoglobin (metHb) or cytochrome c (cyt c) binding to the model lipid membranes composed of zwitterionic lipid phosphatidylcholine (PC) and anionic lipid cardiolipin (CL). Protein association with the lipid vesicles was followed by drastic quenching of SQ-1 fluorescence. The observed spectral changes were suppressed in the presence of free radical scavengers, butylated hydroxytoluene (BHT) and thiourea (TM) suggesting that SQ-1 decolorization can be attributed to its reactions with lipid radicals.     

磷脂跨膜交换机制的研究

膜间磷脂交换是一项重要的生理活动, 其对药物运输及膜功能研究有重要意义. 本文用石英晶体微天平及耗散系数测试仪研究囊泡与囊泡、囊泡与支撑膜间磷脂交换行为, 荧光光谱仪用来测量膜表面电性与膜组分对磷脂交换的影响. 实验结果表明: 磷脂跨膜交换速率与交换时间成反比, 膜表面异电性磷脂的增加会加速膜内相互作用和磷脂跨膜交换速率, 以及改变膜表面组分会对囊泡与支撑膜间的磷脂交换产生影响. 本文研究有助于加深理解磷脂跨膜交换机制, 并对药学研究提供参考.     

Characterization of dimyristoylphosphatidylcholine liposome aggregates induced by dextran sulfate and La3+ by fluorescence spectroscopy

The addition of dextran sulfate (DS) to DMPC vesicles in the presence of di- and trivalent cations leads to a strong aggregation, resulting in a stack-like arrangement of the opposing membrane surfaces as shown by freeze-fracture electron microscopy. The strong aggregation is connected with a lipid mixing process, especially in the presence of La³⁺ (measured by the NBD/Rh assay). The extent of lipid mixing depends on the molecular weight of DS and size of the DMPC vesicles. Additionally, a decrease in the surface dielectric constant of DMPC vesicles [measured by the emission shift of the fluorescent probe, dansylphosphatidyl-ethanolamine (DPE)] was observed. A direct dependence on the molecular weight (MW) of DS exists: the higher their MW, the higher the blue emission shift of the DPE probe. The results are discussed in terms of the theory proposed by Ohki and Arnold, which connects the decrease of the surface dielectric constant with the interaction parameters of phospholipid membranes.     

Raman spectroscopy study of the interaction of 3,5,3′‐triiodo‐L‐thyronine with phosphatidylglycerol lipid bilayers

A Raman spectroscopic study of anionic model membranes and their structural alterations exerted by a relatively small biomolecule, such as the hormone 3,5,3′‐triiodo‐L‐thyronine (T3), is presented. Spectral differences between pure dipalmitoylphosphatidylglycerol (DPPG) multilamellar vesicles and DPPG–T3 mixture and between pure dilauroylphosphatidylglycerol (DLPG) and DLPG–T3 mixture were evaluated in order to determine the response of lipid membranes in gel and liquid‐crystalline phases to the hormone incorporation. Density functional theory (DFT) calculations support the band analysis of the complex 1150–1050 cm⁻¹ Raman region. Geometry optimizations and vibrational behaviors of a model charged molecule that mimics the phosphatidylglycerol lipid moiety in solvated state were taken into account for the spectral interpretation of this specific region. The anionic nature of the lipid polar head plays an important role in the interaction with the hormone, as is evidenced by the CO and PO₂⁻ stretching bands. In addition, the differential penetration of T3 into the hydrophobic region of the membranes shows to be dependent on the lipid phase. The spectral data were compared with those previously obtained for zwitterionic membranes. Copyright © 2015 John Wiley & Sons, Ltd.     

Hyperviscous diblock copolymer vesicles

Giant vesicles prepared from the diblock copolymer polybutadien-b-polyethyleneoxide (PB-PEO) exhibit a shear surface viscosity, which is about 500 times higher than those found in common phospholipid bilayers. Our result constitutes the first direct measurement of the shear surface viscosity of such polymersomes. At the same time, we measure bending and stretching elastic constants, which fall in the range of values typical for lipid membranes. Pulling out a tether from an immobilized polymersome and following its relaxation back to the vesicle body provides an estimate of the viscous coupling between the two monolayers composing the polymer membrane. The detected intermonolayer friction is about an order of magnitude higher than the characteristic one for phospholipid membranes. Polymersomes are tough vesicles with a high lysis tension. This, together with their robust rheological properties, makes them interesting candidates for a number of technological applications. Received 2 March 2001 and Received in final form 15 February 2002     

Interactions of lactoferricin B derivatives with model cell membrane studied by Raman spectroscopy

In this work, we employed Raman spectroscopy to study the effect of the antimicrobial peptide lactoferricin B (LfB) on model cell membranes. We used two derivatives of LfB (RRWQWRMKKLG and RRWQWR) with broad‐spectrum activity against gram‐positive and gram‐negative bacteria, fungus, viruses and tumors. Raman spectra of the peptides showed no conformational change in the temperature range 4–60 °C. The positions of the amide I and amide III bands suggest that in an aqueous solution these peptides preferentially adopt a random coil‐like conformation. We also investigated the effect the peptides had on the melting behavior of model cell membranes composed of zwitterionic lipid dipalmitoylglycero‐phosphocholine (DPPC) and anionic lipid dipalmitoylglycero‐phosphoglycerol (DPPG). Raman CH stretching bands were used to follow the melting of the lipid vesicles. We found that the melting of DPPC lipid vesicles is not affected by the presence of the peptides, while the presence of the peptides reduced cooperativity of the phase transition for anionic DPPG vesicle, suggesting that both peptides interact strongly and specifically with this model cell membrane composed of anionic lipid. Copyright © 2006 John Wiley & Sons, Ltd.     

荧光光谱对自组装多肽作为药物载体的初步研究

为解决疏水性药物普遍存在的因水中溶解度低而给药困难、生物利用度低的问题,采用了新型两亲性自组装多肽RGA16(Ac-RADAGAGARADAGAGS-NH₂)作为载体包裹和释放疏水性模型药物。以芘为模型疏水性药物,以鸡蛋卵磷脂脂质体模拟细胞膜,通过稳态荧光光谱表征和测定芘的存在形式和浓度。两亲性自组装多肽RGA16能够在水溶液中稳定模型疏水性药物芘的晶体。扫描电镜图像显示多肽RGA16与芘晶体相互吸引,两者形成10 μm以上大小的复合体。在机械搅拌下多肽RGA16与水溶液中的芘相互作用5 d左右形成稳定的胶体混悬液(多肽-芘复合体)。被多肽包裹时,芘以晶体的形式存在。而当与EPC脂质体溶液混合时,芘可从多肽的包裹中以分子形式释放到EPC的双层膜中。芘从自组装多肽所稳定的胶态晶体向EPC脂质体释放的过程采用连续时间扫描稳态荧光光谱加以观察。通过将释放过程中芘单体的荧光强度与标准曲线相比较,确定了特定时间点EPC脂质体中芘的转移量。以上结果表明：该两亲性自组装多肽RGA16具有作为小分子量疏水性药物载体的潜力。     

Vesicles and red blood cells in flow: From individual dynamics to rheology

The rheology of suspensions of soft particles, such as red blood cells, is a long-standing problem in science and engineering due to the complex interplay between deformable microstructure and the macroscale flow. The major challenge stems from the free-boundary nature of the particle interface. Lipid bilayer membranes that envelop cells and vesicles are particularly complex interfaces because of their unusual mechanics: the molecularly thin membrane is a highly-flexible incompressible fluid sheet. As a result, particles made of closed lipid bilayers (red cells and vesicles) can exhibit richer dynamics than would capsules and drops. We overview the key experimental observations and recent advances in the theoretical modeling of the vesicles and red blood cells in flow. To cite this article: P.M. Vlahovska et al., C. R. Physique 10 (2009).     

End-to-end Distance Distribution in Fluorescent Derivatives of Bradykinin in Interaction with Lipid Vesicles

Cellular membranes have relevant roles in processes related to proteases like human kallikreins and cathepsins. As enzyme and substrate may interact with cell membranes and associated co-factors, it is important to take into account the behavior of peptide substrates in the lipid environment. In this paper we report an study based on energy transfer in two bradykinin derived peptides labeled with the donor-acceptor pair Abz/Eddnp (ortho-aminobenzoic acid/N-[2,4-dinitrophenyl]-ethylenediamine). Time-resolved fluorescence experiments were performed in phosphate buffer and in the presence of large unilamelar vesicles of phospholipids, and of micelles of sodium dodecyl sulphate (SDS). The decay kinetics were analyzed using the program CONTIN to obtain end-to-end distance distribution functions f(r). Despite of the large difference in the number of residues the end-to-end distance of the longer peptide (9 amino acid residues) is only 20?% larger than the values obtained for the shorter peptide (5 amino acid residues). The proline residue, in position 4 of the bradykinin sequence promotes a turn in the longer peptide chain, shortening its end-to-end distance. The surfactant SDS has a strong disorganizing effect, substantially broadening the distance distributions, while temperature increase has mild effects in the flexibility of the chains, causing small increase in the distribution width. The interaction with phospholipid vesicles stabilizes more compact conformations, decreasing end-to-end distances in the peptides. Anisotropy experiments showed that rotational diffusion was not severely affected by the interaction with the vesicles, suggesting a location for the peptides in the surface region of the bilayer, a result consistent with small effect of lipid phase transition on the peptides conformations.     

Active fusion and fission processes on a fluid membrane

We investigate the steady states and dynamical instabilities resulting from "particles" depositing on (fusion) and pinching off (fission) a fluid membrane. These particles could be either small lipid vesicles or isolated proteins. In the stable case, such fusion/fission events suppress long wavelength fluctuations of the membrane. In the unstable case, the membrane shoots out long tubular structures reminiscent of endosomal compartments or folded structures which bear a morphological resemblance to internal membranes of the cell.     

Microspectroscopic Study of Liposome-to-cell Interaction Revealed by Förster Resonance Energy Transfer

We report the Förster resonance energy transfer (FRET)-labeling of liposomal vesicles as an effective approach to study in dynamics the interaction of liposomes with living cells of different types (rat hepatocytes, rat bone marrow, mouse fibroblast-like cells and human breast cancer cells) and cell organelles (hepatocyte nuclei). The in vitro experiments were performed using fluorescent microspectroscopic technique. Two fluorescent dyes (DiO as the energy donor and DiI as an acceptor) were preloaded in lipid bilayers of phosphatidylcholine liposomes that ensures the necessary distance between the dyes for effective FRET. The change in time of the donor and acceptor relative fluorescence intensities was used to visualize and trace the liposome-to-cell interaction. We show that FRET-labeling of liposome vesicles allows one to reveal the differences in efficiency and dynamics of these interactions, which are associated with composition, fluidity, and metabolic activity of cell plasma membranes.     

Dynamic domain formation in membranes: Thickness-modulation-induced phase separation

A simple model investigates the amplification of fluctuations on membranes constituted of two lipids having different lengths. Van der Waals and electrostatic interactions across the lipid bilayer result in a destabilization favoring thickness variations of the membrane. Close to spontaneous demixing of the two components, the additional gain in free energy due to thickness undulations shifts the stability boundary which promotes phase separation into domains. Interestingly, this effect can be induced by an applied electric field or membrane potential. In biological systems, the dynamic model presented here indicates that electric fields might be important for controlling phase separation and the formation of domains called rafts.Received: 26 March 2004, Published online: 15 July 2004PACS: 87.16.Dg Membranes, bilayers, and vesicles - 68.35.Rh Phase transitions and critical phenomena - 82.70.Uv Surfactants, micellar solutions, vesicles, lamellae, amphiphilic systems, (hydrophilic and hydrophobic interactions)     

Microscopical investigations of nisin-loaded nanoliposomes prepared by Mozafari method and their bacterial targeting

Nanoencapsulation may improve activity of protein or polypeptide antimicrobials against a variety of microorganisms. In this study, nanoliposomes prepared from different lipids (Phospholipon 90H, Phospholipon 100H, dipalmitoylphosphatidylcholine (DPPC), stearylamine (SA), dicetyl phosphate (DCP) and cholesterol) by a new, non-toxic and scalable method, were tested for their capacity to encapsulate nisin Z and target bacteria (Bacillus subtilis and Pseudomonas aeruginosa). Factors affecting the entrapment efficiency (charge and cholesterol concentration in the vesicles) and stability of nanoliposomes were assessed. The nanoliposomes and their bacterial targeting were visualised, using different microscopes under air and liquid environments. Nisin was entrapped in different nanoliposomes with encapsulation efficiencies (EE) ranging from 12% to 54%. Anionic vesicles possessed the highest EE for nisin while increase in cholesterol content in lipid membranes up to 20% molar ratio resulted in a reduction in EE. Stability of nanoliposome-encapsulated nisin was demonstrated for at least 14 months at 4 °C (DPPC:DCP:CHOL vesicles) and for 12 months at 25 °C (DPPC:SA:CHOL vesicles).     

Organization in lipid membranes containing cholesterol

A fundamental attribute of raft formation in cell membranes is lateral separation of lipids into coexisting liquid phases. Using fluorescence microscopy, we observe spontaneous lateral separation in free-floating giant unilamellar vesicles. We record coexisting liquid domains over a range of composition and temperature significantly wider than previously reported. Furthermore, we establish correlations between miscibility in bilayers and in monolayers. For example, the same lipid mixtures that produce liquid domains in bilayer membranes produce two upper miscibility critical points in the phase diagrams of monolayers.