Membrane Viscosity Research Articles

Using flow to investigate friction and protein transport in lipid membranes.

Many cells display lipid-anchored proteins on their plasma membranes. Fluid flow parallel to the cells may displace these proteins. Examples of this have been observed, but lateral flow transport of membrane proteins has not been widely studied. Measurements of the shear force, membrane friction, and resulting protein velocity will make it possible to determine whether this phenomenon is common and contributes to flow sensing. Our lab has developed a method for measuring flow transport of lipid-anchored proteins in a simplified model system: supported bilayer patches. We show that this method is robust and that the flow mobility observed for lipid-anchored proteins under shear flow depends on the ratio of the hydrodynamic force applied to the protein to the frictional drag provided by the lipid anchor. We show that the expected results are observed when these factors are independently varied. One factor that contributes to membrane drag on lipid-anchored proteins is the interleaflet friction. Interleaflet friction is a fundamental parameter governing membrane mechanics: along with membrane viscosity, it determines the time and energy scales for membrane deformations. However, measurements of this parameter are sparse and have previously been accomplished by a large variety of different experimental techniques, making it difficult to compare them or to identify trends. We adapted a recently developed method to determine how interleaflet friction varies with membrane lipid composition. We show that this method gives reproducible results and can detect changes to interleaflet friction resulting from even small differences in acyl chain structure. In particular, we observe dramatic changes to friction when cholesterol is included in the membrane. We look forward to adapting these techniques to investigate how membrane mechanics impact the mobility of proteins and membranes in living cells.

Disorder within disordered regions of protein-free lipid bilayers.

Optical microscopy-based reports on protein-free lipid bilayers with heterogeneous compositions generally consider observed phase separations as thermodynamic endpoints; while MD simulations indicate kinetic characteristics of domain formations. Here, we study GUVs of POPC:DOPE (9:1) and POPC:DPPC (9:1), labeled with disordered-phase probe Liss-RhodPE (1/1000 w/w), using confocal imaging. In addition to confirming previously well-reported disorder and/or phase-separations in vesicular membranes of above compositions, we report the discovery of non-uniform probe distributions within disordered phases, indicating varying degrees of disorder (VDD) for the same vesicular membrane. We further examine dynamics of probe distributions in VDD regions of vesicular membranes. Regression analysis of time-dependent changes in average pixel intensities of probe molecules (avPIs) in different regions of interest (ROIs) allowed quantitative characterization of local (within ROIs) and global (overall vesicular membrane) motion of probe molecules within disordered phases. Optics-related artifacts (e.g., photobleaching, excitation and/or emission fluctuations) were ruled out by noting negligible variations in whole-vesicle avPIs with time. However, avPIs within certain ROIs showed considerable changes with time. In fact, in the same vesicular membrane, avPIs in different ROIs within disordered phases show (a) linear dependence on time (indicating diffusive motion), (b) dependence on square of time (indicating convective motion), and (c) other dependencies on time (e.g., logarithmic, exponential, power, 1/time etc.) - all clubbed together as non-diffusive-non-convective motions. Thus, we rigorously establish existence of further (at least three) levels of heterogeneity, based on dynamics of probe molecules, within disordered phases of protein-free lipid bilayers. Such heterogeneity has previously been ascribed only to biologically active membranes due to presence of membrane proteins. Interestingly, while diffusive motion may be linked to overall membrane viscosity, convective motion may be linked to concepts such as line-tension resulting from heterogeneous compositions.

Cholesterol as a tool to probe the role of membrane in cochlear hair cell mechanotransduction.

Most mechanically gated channels are sensitive to force translated through the membrane. The lipid bilayer can modulate channel function directly through lipid/protein interactions or indirectly based on membrane mechanical properties. Cholesterol is an important component of the membrane and a major modulator of membrane mechanical properties. While other ion channels and mechanically gated channels can be activated or modulated by changes to the membrane cholesterol, the functional role of membrane cholesterol in regulation of mammalian cochlear mechanotransduction (MET) channels has not been closely investigated. Using whole-cell patch clamping and live-cell fluorescence lifetime imaging (FLIM) of a viscosity-sensitive molecular rotor BODIPY 1c for the first time in the inner ear, we examined the role of membrane cholesterol in modulating the MET response properties of rat cochlear hair cells. Molecular rotors are fluorophores for which the fluorescence lifetime (the average time a fluorophore remains in the excited state) increase with increasing viscosity of their immediate environment. We confirmed extraction of cholesterol, using methyl β cyclodextrin (MβCD), with reduced filipin staining in both inner and outer hair bundles. MβCD results in reversible reduction in fluorescence lifetime in hair bundles, suggesting initial reduction followed by gradual recovery in the stereocilia membrane viscosity. MβCD reversibly increases the channel resting open probability, suggesting that cholesterol depletion increases force transfer to the MET channel. Together this data suggests that the cell membrane is part of the force relay machinery to the MET channel and could possibly interact directly with components of the MET machinery. Further studies are needed to generate causal link between MET channel gating and membrane mechanics.

Collective dynamics in the lipid phases of DMPC/cholesterol bilayers: A neutron spin-echo study.

Cholesterol is essential in cell membranes as it regulates membrane structural and material properties such as phase behavior, elasticity, and viscosity. Cholesterol levels in different organelles vary, from less than 5% in mitochondria to 40% in the plasma membrane of eukaryotic cells. According to an experimental phase diagram of 1,2-dimyristoyl-sn-glycerol-3-phosphocholine (DMPC)/cholesterol mixture based on lateral diffusion measurements from early studies, at the above melting temperature, the system undergoes a phase transition from a liquid-disordered (Ld) phase to a liquid-ordered (Lo) phase with a coexisting phase (Ld+Lo) in between upon increasing cholesterol concentration [1]. Our work attempts to understand the collective dynamics of the mixture in each phase and quantify their behavioral trends. We first performed densitometry to determine the mixture's molecular volume and to correct the melting temperature difference between protiated and deuterated DMPC, which was used in neutron experiments. The molecular volume was then implemented in small angle neutron scattering (SANS) experiments and data analysis to extract the area per lipid (AL) and thickness of the lipid mixture in each phase region. Finally, neutron spin echo (NSE) is used to access the dynamic properties by adopting structural information from SANS. The NSE results show the effective bending modulus and membrane viscosity of the Ld phase scale with AL, as observed in our previous study of mixed lipid membranes [2].

Photo-Activated Ratiometric Fluorescent Indicator for Real-Time and Visual Detection of Plasma Membrane Homeostasis.

Development of an activated ratiometric indicator that is specific to plasma membrane (PM) viscosity exhibits great application prospects in disease diagnosis and treatment but remains a great challenge. Herein, a photo-activated fluorescent probe (CQ-IC) was designed and prepared tactfully, which could analyze and real-time monitor the microenvironmental homeostasis of the PM based on a two-channel ratiometric imaging model. Interestingly, upon light irradiation, CQ-IC generates reactive oxygen species and thus increases the cellular viscosity, which increases two emission peaks at 480 and 610 nm. This work would propose a new strategy to sensor PM homeostasis and effectively guide the treatment of viscosity-related diseases among various physiological and pathological processes.

Energy Dissipation in the Human Red Cell Membrane

The membrane of the human red cell consists of a lipid bilayer and a so-called membrane skeleton attached on the cytoplasmic side of the bilayer. Upon the deformation of red cells, energy is dissipated in their cytoplasm and their membrane. As to the membrane, three contributions can be distinguished: (i) A two-dimensional shear deformation with the membrane viscosity as the frictional parameter; (ii) A motion of the membrane skeleton relative to the bilayer; (iii) A relative motion of the two monolayers of the bilayer. The frictional parameter in contributions (ii) and (iii) is a frictional coefficient specific for the respective contribution. This perspective describes the history up to recent advances in the knowledge of these contributions. It reviews the mechanisms of energy dissipation on a molecular scale and suggests new ones, particularly for the first contribution. It proposes a parametric fitting expected to shed light on the discrepant values found for the membrane viscosity by different experimental approaches. It proposes strategies that could allow the determination of the frictional coefficients pertaining to the second and the third contribution. It highlights the consequences characteristic times have on the state of the red cell membrane in circulation as well as on the adaptation of computer models to the red cell history in an in vitro experiment.

Effects of crowding on the diffusivity of membrane adhered particles.

The lateral diffusion of cell membrane inclusions, such as integral membrane proteins and bound receptors, drives critical biological processes, including the formation of complexes, cell-cell signaling, and membrane trafficking. These diffusive processes are complicated by how concentrated, or "crowded", the inclusions are, which can occupy between 30-50% of the area fraction of the membrane. In this work, we elucidate the effects of increasing concentration of model membrane inclusions in a free-standing artificial cell membrane on inclusion diffusivity and the apparent viscosity of the membrane. By multiple particle tracking of fluorescent microparticles covalently tethered to the bilayer, we show the transition from expected Brownian dynamics, which accurately measure the membrane viscosity, to subdiffusive behavior with decreased diffusion coefficient as the particle area fraction increases from 1% to around 30%, approaching physiological levels of crowding. At high crowding, the onset of non-Gaussian behavior is observed. Using hydrodynamic models relating the 2D diffusion coefficient to the viscosity of a membrane, we determine the apparent viscosity of the bilayer from the particle diffusivity and show an increase in the apparent membrane viscosity with increasing particle area fraction. However, the scaling of this increase is in contrast with the behavior of monolayer inclusion diffusion and bulk suspension rheology. These results demonstrate that physiological levels of model membrane crowding nontrivially alter the dynamics and apparent viscosity of the system, which has implications for understanding membrane protein interactions and particle-membrane transport processes.

Fatty Acid Composition of Comamonas testosteroni under Hexachlorobenzene Loading Conditions

Changes in the lipid composition in bacterial membranes are considered to be the most important adaptation mechanisms to adverse chemical factors. The aim of the study was to compare the hexachlorobenzene effects on the fatty acid composition of total lipids Comamonas testosteroni. Methods. The study was performed with C. testosteroni UCM B-400 and B-401, B-213 strains. Bacteria were grown in the Luria-Bertrani (LB) liquid medium containing 10 and 20 mg/L of hexachlorobenzene (HCB). After cultivation, the biomass was separated by centrifugation and the fatty acid composition of total lipids was determined through analyzing its methyl esters. To assess the cell membrane properties, such parameters as the lipid unsaturation index, the average carbon chain length of fatty acids, and the membrane viscosity index were determined. Results. In the fatty acids spectrum of C. testosteroni B-400 after cultivation in a medium containing 20 mg/L of HCB, the contents of unsaturated hexadecenoic (C16:1) and octadecenoic (C18:1) acids were lower by 10.6 and 5.5%, respectively, and that of saturated hexadecanoic (C16:0) acid was higher by 8.4%, compared to the control. The fatty acid composition of C. testosteroni B-401 was more stable compared to strain B-400. Collection strain C. testosteroni B-213 compared to strains isolated from soil with high HCB load, in the presence of 10 and 20 mg/L of HCB had the highest relative content of saturated hexadecanoic acid (C16:0) up to 38.33—40.7%. Unsaturated octadecenoic acid decreased at the doses 10 and 20 mg/L to 1.5—2% compared to the control. In all strains under the HCB impact, there was an increase in the relative content of C17-cyclopropanoic acid compared to control variants. Conclusions. C. testosteroni UCM B-400, B-401, and B-213 bacteria under cultivation conditions in HCB-containing medium, decreasing the degree of lipid unsaturation and increasing the relative content of C17-cyclopropanoic acid can be considered as the main mechanisms regulating the cytoplasmic membrane fluidity; the displaying of these protective reactions had a strain trait and did not depend on the adaptation in natural isolating places.

Holmium Complex with Phospholipids as 1H NMR Relaxational Sensor of Temperature and Viscosity.

The sensitivity of Ho–phospholipid complexes to changes in the membrane viscosity of liposomes was checked. An increase in viscosity was observed for DPPC and DMPC near the phase-transition temperature. Ho–phospholipid complexes could be used as sensors of local membrane viscosity in NMR and MRI technologies.

Heterogeneous Nanostructures Cause Anomalous Diffusion in Lipid Monolayers.

The diffusion and mobility in biomembranes are crucial for various cell functions; however, the mechanisms involved in such processes remain ambiguous due to the complex membrane structures. Herein, we investigate how the heterogeneous nanostructures cause anomalous diffusion in dipalmitoylphosphatidylcholine (DPPC) monolayers. By identifying the existence of condensed nanodomains and clarifying their impact, our findings renew the understanding of the hydrodynamic description and the statistical feature of the diffusion in the monolayers. We find a universal characteristic of the multistage mean square displacement (MSD) with an intermediate crossover, signifying two membrane viscosities at different scales: the short-time scale describes the local fluidity and is independent of the nominal DPPC density, and the long-time scale represents the global continuous phase taking into account nanodomains and increases with DPPC density. The constant short-time viscosity reflects a dynamic equilibrium between the continuous fluid phase and the condensed nanodomains in the molecular scale. Notably, we observe an "anomalous yet Brownian" phenomenon exhibiting an unusual double-peaked displacement probability distribution (DPD), which is attributed to the net dipolar repulsive force from the heterogeneous nanodomains around the microdomains. The findings provide physical insights into the transport of membrane inclusions that underpin various biological functions and drug deliveries.

Artemisinin Targets Transcription Factor PDR1 and Impairs Candida glabrata Mitochondrial Function.

A limited number of antifungal drugs, the side-effect of clinical drugs and the emergence of resistance create an urgent need for new antifungal treatment agents. High-throughput drug screening and in-depth drug action mechanism analyzation are needed to address this problem. In this study, we identified that artemisinin and its derivatives possessed antifungal activity through a high-throughput screening of the FDA-approved drug library. Subsequently, drug-resistant strains construction, a molecular dynamics simulation and a transcription level analysis were used to investigate artemisinin’s action mechanism in Candida glabrata. Transcription factor pleiotropic drug resistance 1 (PDR1) was an important determinant of artemisinin’s sensitivity by regulating the drug efflux pump and ergosterol biosynthesis pathway, leading to mitochondrial dysfunction. This dysfunction was shown by a depolarization of the mitochondrial membrane potential, an enhancement of the mitochondrial membrane viscosity and an upregulation of the intracellular ROS level in fungi. The discovery shed new light on the development of antifungal agents and understanding artemisinin’s action mechanism.

Modeling and optimization of neodymium ion separation by liquid membrane using Artificial Neural Network coupled with Genetic Algorithm

The synergistic effects of two or more carrier species significantly intensify the performance of liquid membranes. However, this process is difficult to model as different carrier species make different complexes with permeating species. Artificial Neural Network (ANN) based modeling is helpful in this case. First number of neurons in the hidden layer was optimized to reduce mean squared error (MSE). Minimum MSE (10.96) was achieved with ten neurons. Simulations showed that neodymium transport initially increased with increasing carrier species TOPO concentration but later decreased due to increased viscosity of liquid membrane. Similarly, higher neodymium transport was found at a moderate concentration of H2SO4 in receiving phase and a low concentration of HCl in the feed phase. Trained ANN was coupled with the Genetic Algorithm to determine the optimum operating parameters in their given range. 0.12 M TOPO concentration in the liquid membrane, 4.2 M of H2SO4 concentration in the receiving phase, and 0.5 M of HCl concentration in the feed phase were optimum values of the operating parameters to achieve 100% neodymium transport through the liquid membrane in 360 min operation.

Development of resistance to 5-fluorouracil affects membrane viscosity and lipid composition of cancer cells

The investigations reported here were designed to determine whether the bulk plasma membrane is involved in mechanisms of acquired resistance of colorectal cancer cells to 5-fluorouracil (5-FU). Fluorescence lifetime imaging microscopy (FLIM) of live cultured cells stained with viscosity-sensitive probe BODIPY 2 was exploited to non-invasively assess viscosity in the course of treatment and adaptation to the drug. In parallel, lipid composition of membranes was examined with the time-of-flight secondary ion mass spectrometry (ToF-SIMS). Our results showed that a single treatment with 5-FU induced only temporal changes of viscosity in 5-FU sensitive cells immediately after adding the drug. Acquisition of chemoresistance was accompanied by persistent increase of viscosity, which was preserved upon treatment without any changes. Lipidomic analysis revealed that the resistant cells had a lower level of monounsaturated fatty acids and increased sphingomyelin or decreased phosphatidylcholine in their membranes, which partly explain increase of the viscosity. Thus, we propose that a high membrane viscosity mediates the acquisition of resistance to 5-FU.

Tank-treading dynamics of red blood cells in shear flow: On the membrane viscosity rheology

In this article, extensive three-dimensional simulations are conducted for tank-treading (TT) red blood cells (RBCs) in shear flow with different cell viscous properties and flow conditions. Apart from recent numerical studies on TT RBCs, this research considers the uncertainty in cytoplasm viscosity, covers a more complete range of shear flow situations of available experiments, and examines the TT behaviors in more details. Key TT characteristics, including the rotation frequency, deformation index, and inclination angle, are compared with available experimental results of similar shear flow conditions. Fairly good simulation-experiment agreements for these parameters can be obtained by adjusting the membrane viscosity values; however, different rheological relationships between the membrane viscosity and the flow shear rate are noted for these comparisons: shear thinning from the TT frequency, Newtonian from the inclination angle, and shear thickening from the cell deformation. Previous studies claimed a shear-thinning membrane viscosity model based on the TT frequency results; however, such a conclusion seems premature from our results and more carefully designed and better controlled investigations are required for the RBC membrane rheology. In addition, our simulation results reveal complicate RBC TT features and such information could be helpful for a better understanding of in vivo and in vitro RBC dynamics.

An isogeometric finite element formulation for boundary and shell viscoelasticity based on a multiplicative surface deformation split

AbstractThis work presents a numerical formulation to model isotropic viscoelastic material behavior for membranes and thin shells. The surface and the shell theory are formulated within a curvilinear coordinate system, which allows the representation of general surfaces and deformations. The kinematics follow from Kirchhoff–Love theory and the discretization makes use of isogeometric shape functions. A multiplicative split of the surface deformation gradient is employed, such that an intermediate surface configuration is introduced. The surface metric and curvature of this intermediate configuration follow from the solution of nonlinear evolution laws—ordinary differential equations—that stem from a generalized viscoelastic solid model. The evolution laws are integrated numerically with the implicit Euler scheme and linearized within the Newton–Raphson scheme of the nonlinear finite element framework. The implementation of membrane and bending viscosity is verified with the help of analytical solutions and shows ideal convergence behavior. The chosen numerical examples capture large deformations and typical viscoelasticity behavior, such as creep, relaxation, and strain rate dependence. It is also shown that the proposed formulation can be straightforwardly applied to model boundary viscoelasticity of 3D bodies.

Mitochondria-Assisted Photooxidation to Track Singlet Oxygen at Homeostatic Membrane Microviscosity.

Using intracellular-controlled photochemistry to track dynamic organelle processes is gaining attention due to its broad applications. However, most of the employed molecular probes usually require toxic photosensitizers and complex bioanalytical protocols. Here, the synthesis and performance of two new subcellular probes (MitoT1 and MitoT2) are described. The probes undergo photooxidation in the damaged tissue of zebrafish, a model system for tissue regeneration studies. Using high-resolution confocal microscopy and fluorescence spectroscopy, we combine the mentioned photoinduced interconversion at the homeostatic membrane viscosity to track singlet oxygen activity selectively. The continuous and real-time biosensing method reported here provides a new approach for simultaneously detecting endogenous singlet oxygen and viscosity status.

P-198 An analysis of the size of micro pronucleus in 2.1 pronuclear zygotes by using time-lapse images

Abstract Study question Is it possible to determine the difference between 2.1 pronuclear (2.1PN) zygotes and tripronuclear (3PN) zygotes from time-lapse images? Summary answer A pronucleus of less than 15 μm in diameter can be considered the micro pronucleus (micro PN), and it is possible to classify 2.1PN zygotes. What is known already 2.1PN zygotes are defined as zygotes with two pronuclei and one smaller pronucleus. Capalbo et al. (2017) reported that most of the 2.1PN-derived blastocysts were diploid by preimplantation genetic testing for aneuploidy (PGT-A), including single-nucleotide polymorphisms (SNPs) analysis. Thus, the treatment with 2.1 PN zygotes should be performed with chromosome testing. In Japan, where PGT-A is not available in principle, 2.1PN zygotes are rarely used in the embryo transfer. On the other hand, the size of the micro pronucleus in 2.1PN zygotes has not been clearly defined, and it is difficult to determine differences between 2.1PN and 3PN zygotes. Study design, size, duration The study was performed retrospectively on 2463 cycles of in vitro fertilization (IVF) conducted at our clinic between August 2020 and December 2021. A total of 3073 embryos underwent conventional-IVF (c-IVF) or intracytoplasmic sperm injection (ICSI) and were cultured in the time-lapse incubator, of which 221 zygotes with three pronuclei were used in the study. Participants/materials, setting, methods The diameter of the three PNs at one hour before syngamy from time-lapse images; 2.1 PN and 3PN zygotes were classified in the report by Capalbo et al. (2017). The age of the patients and the method of insemination between the groups were compared, and the diameter of the micro PN was analysed. Moreover, logistic regression analysis was performed to investigate the predictor of 2.1PN zygotes from the morphological characteristics of oocytes at ICSI. Main results and the role of chance The mean age of each patient was 42.9 years for 2.1PN zygotes and 39.8 years for 3PN zygotes, significantly higher for 2.1PN zygotes (P =0.003). On the other hand, when comparing the stage of oocyte maturation at the time of oocyte retrieval, there was no significant difference (P =0.749). According to the insemination method, the incidence of 2.1PN zygotes was significantly higher in ICSI (including rescue-ICSI) compared to c-IVF: 32.9% [95%CI: 22.5-44.6%] vs 2.4% [95%CI: 0.1-12.9%] (P <0.001). In terms of ICSI-derived zygotes, the mean age was also significantly higher for 2.1PN zygotes compared to 3PN zygotes: 43.3 years vs. 40.9 years (P =0.03). The diameter of micro PNs calculated using the receiver operating characteristics (ROC) curve from the measurements of the diameter was less than 15 μm (AUC [95%CI]: AUC=0.988 [0.975-1.00]). Logistic regression analysis using age, position of the oocyte spindle at ICSI, cytoplasmic viscosity, and condition of the cell membrane as explanatory variables revealed a significant difference only in age (P =0.0154, odds ratio [95%CI]: 1.18 [1.03-1.35]) and no statistically significant oocyte morphological characteristics. Limitations, reasons for caution In this study, we have not investigated whether 2.1 PN zygotes become blastocysts. It will be necessary to further examine the criteria for 2.1PN along with chromosome testing to investigate the use of 2.1PN-derived blastocysts. Wider implications of the findings A pronucleus of less than 15 μm in diameter can be considered a micro PN. Compared to 3PN zygotes, 2.1PN zygotes were more frequently observed in older patients and in ICSI-derived zygotes. However, it is difficult to predict the incidence of 2.1PN zygotes from the oocytes’ morphological characteristics at ICSI. Trial registration number not applicable

Assessing hypoxic damage to placental trophoblasts by measuring membrane viscosity of extracellular vesicles.

As highly sophisticated intercellular communication vehicles in biological systems, extracellular vesicles (EVs) have been investigated as both promising liquid biopsy-based disease biomarkers and drug delivery carriers. Despite tremendous progress in understanding their biological and physiological functions, mechanical characterization of these nanoscale entities remains challenging due to the limited availability of proper techniques. Especially, whether damage to parental cells can be reflected by the mechanical properties of their EVs remains unknown. In this study, we characterized membrane viscosities of different types of EVs collected from primary human trophoblasts (PHTs), including apoptotic bodies, microvesicles and small extracellular vesicles, using fluorescence lifetime imaging microscopy (FLIM). The biochemical origin of EV membrane viscosity was examined by analyzing their phospholipid composition, using mass spectrometry. We found that different EV types derived from the same cell type exhibit different membrane viscosities. The measured membrane viscosity values are well supported by the lipidomic analysis of the phospholipid compositions. We further demonstrate that the membrane viscosity of microvesicles can faithfully reveal hypoxic injury of the human trophoblasts. More specifically, the membrane of PHT microvesicles released under hypoxic condition is less viscous than its counterpart under standard culture condition, which is supported by the reduction in the phosphatidylethanolamine-to-phosphatidylcholine ratio in PHT microvesicles. Our study suggests that biophysical properties of released trophoblastic microvesicles can reflect cell health. Characterizing EV's membrane viscosity may pave the way for the development of new EV-based clinical applications.

Predictive modeling and experimental implementation of organic acids in stream recovery by reactive extraction in membrane contactors

• Substantial reduction of acid accumulation with the coupled process. • Great viscosity variations due to the transfer. • Viscosity changes successfully taken into account to describe kinetics. • Model-based generalization for process guidelines. Membrane-based reactive extraction as an in situ product recovery technique is a promising strategy for process intensification, in particular in the case of the bioproduction of organic acids. Reactive extraction allows a high selectivity for the extraction of the targeted acid and the microporous membrane keeps biocatalysts in the aqueous broth while implementing a large liquid–liquid surface area and ensuring a dispersion-free contact, without problems of emulsion formation. This paper deals specifically with the extraction of biobased 3-hydroxypropionic acid using tri- n -octylamine in n -decanol. In order to maintain an effective driving force for 3-HP transfer into the organic phase, this latter was continuously regenerated by recovering the acid in a back-extraction aqueous phase, giving a complete pertraction process. A mass transfer model for this process was developed. It is based on the boundary layer theory and takes into account chemical and physical equilibria of complexation/dissociation and partitioning, species diffusion in the membrane pores and viscosity variations in the organic phase. Viscosity highly depends on acid concentration, increasing up to 50% when 3-HP concentration reaches 28 g.L -1 . Thus, it was possible to predict different experimental results with R 2 ≥ 0.99, totally neglecting chemical kinetics and interfacial resistance for both extraction and back-extraction steps. The model allows the prediction of extraction kinetics with (1) fixed initial concentrations and (2) with gradual 3-HP feed (mimicking a bioconversion) in transient and steady states coupled with back-extraction (globally also called pertraction). Model based analysis of mass transfer mechanisms led to the construction of a nomogram giving 3-HP stationary concentration in the case of a typical production rate, enabling for example a rapid organic phase selection or membrane sizing.

A vesicle microrheometer for high-throughput viscosity measurements of lipid and polymer membranes

Viscosity is a key property of cell membranes that controls mobility of embedded proteins and membrane remodeling. Measuring it is challenging because existing approaches involve complex experimental designs and/or models, and the applicability of some methods is limited to specific systems and membrane compositions. As a result there is scarcity of systematic data, and the reported values for membrane viscosity vary by orders of magnitude for the same system. Here, we show how viscosity of membranes can be easily obtained from the transient deformation of giant unilamellar vesicles. The approach enables a noninvasive, probe-independent, and high-throughput measurement of the viscosity of membranes made of lipids or polymers with a wide range of compositions and phase state. Using this novel method, we have collected a significant amount of data that provides insights into the relation between membrane viscosity, composition, and structure.