Electrodeformation of multi-bilayer spherical concentric membranes by AC electric fields

NASA Astrophysics Data System (ADS)

Lira-Escobedo, J.; Arauz-Lara, J.; Aranda-Espinoza, H.; Adlerz, K.; Viveros-Mendez, P. X.; Aranda-Espinoza, S.

2017-09-01

It is now well established that external stresses alter the behaviour of cells, where such alterations can be as profound as changes in gene expression. A type of stresses of particular interest are those due to alternating-current (AC) electric fields. The effect of AC fields on cells is still not well understood, in particular it is not clear how these fields affect the cell nucleus and other organelles. Here, we propose that one possible mechanism is through the deformation of the membranes. In order to investigate the effect of AC fields on the morphological changes of the cell organelles, we modelled the cell as two concentric bilayer membranes. This model allows us to obtain the deformations induced by the AC field by balancing the elastic energy and the work done by the Maxwell stresses. Morphological phase diagrams are obtained as a function of the frequency and the electrical properties of the media and membranes. We demonstrate that the organelle shapes can be changed without modifying the shape of the external cell membrane and that the organelle deformation transitions can be used to measure, for example, the conductivity of the nucleus.

Myosin IIA interacts with the spectrin-actin membrane skeleton to control red blood cell membrane curvature and deformability.

PubMed

Smith, Alyson S; Nowak, Roberta B; Zhou, Sitong; Giannetto, Michael; Gokhin, David S; Papoin, Julien; Ghiran, Ionita C; Blanc, Lionel; Wan, Jiandi; Fowler, Velia M

2018-05-08

The biconcave disk shape and deformability of mammalian RBCs rely on the membrane skeleton, a viscoelastic network of short, membrane-associated actin filaments (F-actin) cross-linked by long, flexible spectrin tetramers. Nonmuscle myosin II (NMII) motors exert force on diverse F-actin networks to control cell shapes, but a function for NMII contractility in the 2D spectrin-F-actin network of RBCs has not been tested. Here, we show that RBCs contain membrane skeleton-associated NMIIA puncta, identified as bipolar filaments by superresolution fluorescence microscopy. MgATP disrupts NMIIA association with the membrane skeleton, consistent with NMIIA motor domains binding to membrane skeleton F-actin and contributing to membrane mechanical properties. In addition, the phosphorylation of the RBC NMIIA heavy and light chains in vivo indicates active regulation of NMIIA motor activity and filament assembly, while reduced heavy chain phosphorylation of membrane skeleton-associated NMIIA indicates assembly of stable filaments at the membrane. Treatment of RBCs with blebbistatin, an inhibitor of NMII motor activity, decreases the number of NMIIA filaments associated with the membrane and enhances local, nanoscale membrane oscillations, suggesting decreased membrane tension. Blebbistatin-treated RBCs also exhibit elongated shapes, loss of membrane curvature, and enhanced deformability, indicating a role for NMIIA contractility in promoting membrane stiffness and maintaining RBC biconcave disk cell shape. As structures similar to the RBC membrane skeleton exist in many metazoan cell types, these data demonstrate a general function for NMII in controlling specialized membrane morphology and mechanical properties through contractile interactions with short F-actin in spectrin-F-actin networks.

[Establishment and application of mechanical strain loading system of multi-channel cells].

PubMed

Li, Yongming; Wang, Hua; Zhang, Xiaodong; Tang, Lin

2012-02-01

Based on single-chip microcomputer, we have established a mechanical strain loading system with multi-channel to study the biological behavior of cultured cells in vitro under mechanical strain. We developed a multi-channel cell strain loading device controlled by single-chip microcomputer. We controlled the vacuum pump with vacuum chamber to make negative pressure changing periodically in the vacuum chamber. The tested cells were seeded on the surface of an elastic membrane mounted on the vacuum chamber, and could be strained or relaxed by cyclic pressure. Since the cells are attached to the surface of the membrane, they presumably experience the same deformation as that was applied to the membrane. The system was easy to carry and to operate, with deformation rate (1%-21%) and frequency (0-0. 5Hz) which could be adjusted correctly according to experimental requirement, and could compare different deformation rate of three channels at the same time. The system ran stably and completely achieved design aims, and provided a method to study the biological behavior of cultured cells attached to the surface of the elastic membrane under mechanical strain in vitro.

Deformation of red blood cells using acoustic radiation forces

PubMed Central

Mishra, Puja; Hill, Martyn; Glynne-Jones, Peter

2014-01-01

Acoustic radiation forces have been used to manipulate cells and bacteria in a number of recent microfluidic applications. The net force on a cell has been subject to careful investigation over a number of decades. We demonstrate that the radiation forces also act to deform cells. An ultrasonic standing wave field is created in a 0.1 mm glass capillary at a frequency of 7.9 MHz. Using osmotically swollen red-blood cells, we show observable deformations up to an aspect ratio of 1.35, comparable to deformations created by optical tweezing. In contrast to optical technologies, ultrasonic devices are potentially capable of deforming thousands of cells simultaneously. We create a finite element model that includes both the acoustic environment of the cell, and a model of the cell membrane subject to forces resulting from the non-linear aspects of the acoustic field. The model is found to give reasonable agreement with the experimental results, and shows that the deformation is the result of variation in an acoustic force that is directed outwards at all points on the cell membrane. We foresee applications in diagnostic devices, and in the possibility of mechanically stimulating cells to promote differentiation and physiological effects. PMID:25379070

Investigation of membrane mechanics using spring networks: application to red-blood-cell modelling.

PubMed

Chen, Mingzhu; Boyle, Fergal J

2014-10-01

In recent years a number of red-blood-cell (RBC) models have been proposed using spring networks to represent the RBC membrane. Some results predicted by these models agree well with experimental measurements. However, the suitability of these membrane models has been questioned. The RBC membrane, like a continuum membrane, is mechanically isotropic throughout its surface, but the mechanical properties of a spring network vary on the network surface and change with deformation. In this work spring-network mechanics are investigated in large deformation for the first time via an assessment of the effect of network parameters, i.e. network mesh, spring type and surface constraint. It is found that a spring network is conditionally equivalent to a continuum membrane. In addition, spring networks are employed for RBC modelling to replicate the optical tweezers test. It is found that a spring network is sufficient for modelling the RBC membrane but strain-hardening springs are required. Moreover, the deformation profile of a spring network is presented for the first time via the degree of shear. It is found that spring-network deformation approaches continuous as the mesh density increases. Copyright © 2014 Elsevier B.V. All rights reserved.

Refractive index tomograms and dynamic membrane fluctuations of red blood cells from patients with diabetes mellitus.

PubMed

Lee, SangYun; Park, HyunJoo; Kim, Kyoohyun; Sohn, YongHak; Jang, Seongsoo; Park, YongKeun

2017-04-21

In this paper, we present the optical characterisations of diabetic red blood cells (RBCs) in a non-invasive manner employing three-dimensional (3-D) quantitative phase imaging. By measuring 3-D refractive index tomograms and 2-D time-series phase images, the morphological (volume, surface area and sphericity), biochemical (haemoglobin concentration and content) and mechanical (membrane fluctuation) parameters were quantitatively retrieved at the individual cell level. With simultaneous measurements of individual cell properties, systematic correlative analyses on retrieved RBC parameters were also performed. Our measurements show there exist no statistically significant alterations in morphological and biochemical parameters of diabetic RBCs, compared to those of healthy (non-diabetic) RBCs. In contrast, membrane deformability of diabetic RBCs is significantly lower than that of healthy, non-diabetic RBCs. Interestingly, non-diabetic RBCs exhibit strong correlations between the elevated glycated haemoglobin in RBC cytoplasm and decreased cell deformability, whereas diabetic RBCs do not show correlations. Our observations strongly support the idea that slow and irreversible glycation of haemoglobin and membrane proteins of RBCs by hyperglycaemia significantly compromises RBC deformability in diabetic patients.

Lipid membrane-mediated attraction between curvature inducing objects

NASA Astrophysics Data System (ADS)

van der Wel, Casper; Vahid, Afshin; Šarić, Anđela; Idema, Timon; Heinrich, Doris; Kraft, Daniela J.

2016-09-01

The interplay of membrane proteins is vital for many biological processes, such as cellular transport, cell division, and signal transduction between nerve cells. Theoretical considerations have led to the idea that the membrane itself mediates protein self-organization in these processes through minimization of membrane curvature energy. Here, we present a combined experimental and numerical study in which we quantify these interactions directly for the first time. In our experimental model system we control the deformation of a lipid membrane by adhering colloidal particles. Using confocal microscopy, we establish that these membrane deformations cause an attractive interaction force leading to reversible binding. The attraction extends over 2.5 times the particle diameter and has a strength of three times the thermal energy (-3.3 kBT). Coarse-grained Monte-Carlo simulations of the system are in excellent agreement with the experimental results and prove that the measured interaction is independent of length scale. Our combined experimental and numerical results reveal membrane curvature as a common physical origin for interactions between any membrane-deforming objects, from nanometre-sized proteins to micrometre-sized particles.

Wrinkling of a spherical lipid interface induced by actomyosin cortex

NASA Astrophysics Data System (ADS)

Ito, Hiroaki; Nishigami, Yukinori; Sonobe, Seiji; Ichikawa, Masatoshi

2015-12-01

Actomyosin actively generates contractile forces that provide the plasma membrane with the deformation stresses essential to carry out biological processes. Although the contractile property of purified actomyosin has been extensively studied, to understand the physical contribution of the actomyosin contractile force on a deformable membrane is still a challenging problem and of great interest in the field of biophysics. Here, we reconstitute a model system with a cell-sized deformable interface that exhibits anomalous curvature-dependent wrinkling caused by the actomyosin cortex underneath the spherical closed interface. Through a shape analysis of the wrinkling deformation, we find that the dominant contributor to the wrinkled shape changes from bending elasticity to stretching elasticity of the reconstituted cortex upon increasing the droplet curvature radius of the order of the cell size, i.e., tens of micrometers. The observed curvature dependence is explained by the theoretical description of the cortex elasticity and contractility. Our present results provide a fundamental insight into the deformation of a curved membrane induced by the actomyosin cortex.

Laser-induced surface deformation microscope for the study of the dynamic viscoelasticity of plasma membrane in a living cell.

PubMed

Morisaku, Toshinori; Yui, Hiroharu

2018-05-15

A laser-induced surface deformation (LISD) microscope is developed and applied to measurement of the dynamic relaxation responses of the plasma membrane in a living cell. A laser beam is tightly focused on an optional area of cell surface and the focused light induces microscopic deformation on the surface via radiation pressure. The LISD microscope not only allows non-contact and destruction-free measurement but provides power spectra of the surface responses depending on the frequency of the intensity of the laser beam. An optical system for the LISD is equipped via a microscope, allowing us to measure the relaxation responses in sub-cellular-sized regions of the plasma membrane. In addition, the forced oscillation caused by the radiation pressure for surface deformation extends the upper limit of the frequency range in the obtained power spectra to 106 Hz, which enables us to measure relaxation responses in local regions within the plasma membrane. From differences in power-law exponents at higher frequencies, it is realized that a cancerous cell obeys a weaker single power-law than a normal fibroblast cell. Furthermore, the power spectrum of a keratinocyte cell obeys a power-law with two exponents, indicating that alternative mechanical models to a conventional soft glassy rheology model (where single power-laws explain cells' responses below about 103 Hz) are needed for the understanding over a wider frequency range. The LISD microscope would contribute to investigation of microscopic cell rheology, which is important for clarifying the mechanisms of cell migration and tissue construction.

Modeling 3-D deformation of outer hair cells and their production of the active force in the cochlea.

PubMed

Spector, A A; Ameen, M; Schmiedt, R A

2002-10-01

We analyze the deformation of the outer hair cell and its production of active force under physiological conditions. The active force has two components. One results from the strain caused by loading in the organ of Corti in the cochlea and depends on the level of the acoustic signal; the other is related to the intrinsic active properties of the cell membrane. We demonstrate our approach by considering, as a basic model of an outer hair cell in the organ of Corti, a cylindrical shell that is filled with an incompressible fluid and located between two planes that move relative to each other. These planes represent the basilar membrane and tectorial membrane complexes. We show that the deformed state of the cell has a 3-D nature, including bending and twisting components. This is different from the experimental conditions in which the active force is usually measured. We estimate the active force as a function of the relative position of the planes, angle of the cell's inclination, and the cell length.

Microconfined flow behavior of red blood cells.

PubMed

Tomaiuolo, Giovanna; Lanotte, Luca; D'Apolito, Rosa; Cassinese, Antonio; Guido, Stefano

2016-01-01

Red blood cells (RBCs) perform essential functions in human body, such as gas exchange between blood and tissues, thanks to their ability to deform and flow in the microvascular network. The high RBC deformability is mainly due to the viscoelastic properties of the cell membrane. Since an impaired RBC deformability could be found in some diseases, such as malaria, sickle cell anemia, diabetes and hereditary disorders, there is the need to provide further insight into measurement of RBC deformability in a physiologically relevant flow field. Here, RBCs deformability has been studied in terms of the minimum apparent plasma-layer thickness by using high-speed video microscopy of RBCs flowing in cylindrical glass capillaries. An in vitro systematic microfluidic investigation of RBCs in micro-confined conditions has been performed, resulting in the determination of the RBCs time recovery constant, RBC volume and surface area and RBC membrane shear elastic modulus and surface viscosity. It has been noticed that the deformability of RBCs induces cells aggregation during flow in microcapillaries, allowing the formation of clusters of cells. Overall, our results provide a novel technique to estimate RBC deformability and also RBCs collective behavior, which can be used for the analysis of pathological RBCs, for which reliable quantitative methods are still lacking. Copyright © 2015 IPEM. Published by Elsevier Ltd. All rights reserved.

Revealing the cell-material interface with nanometer resolution by FIB-SEM

PubMed Central

Santoro, Francesca; Zhao, Wenting; Joubert, Lydia-Marie; Duan, Liting; Schnitker, Jan; van de Burgt, Yoeri; Lou, Hsin-Ya; Liu, Bofei; Salleo, Alberto; Cui, Lifeng; Cui, Yi; Cui, Bianxiao

2018-01-01

The interface between cells and non-biological surfaces regulates cell attachment, chronic tissue responses, and ultimately the success of medical implants or biosensors. Clinical and laboratory studies show that topological features of the surface profoundly influences cellular responses, e.g. titanium surfaces with nano- and microtopographical structures enhance osteoblast attachment and host-implant integration as compare to smooth surface. To understand how cells and tissues respond to different topographical features, it is of critical importance to directly visualize the cell-materials interface at the relevant nanometer length scale. Here, we present a new method for in situ examination of the cell-to-material interface at any desired location, based on focused-ion beam milling and scanning electron microscopy imaging (FIB-SEM) to resolve the cell membrane-to-material interface with 10 nm resolution. By examining how cell membranes interact with topographical features such as nanoscale protrusions or invaginations, we discovered that the cell membrane readily deforms inward and wraps around protruding structures, but hardly deforms outward to contour invaginating structures. This asymmetric membrane response (inward vs. outward deformation) causes the cleft width between the cell membrane and the nanostructure surface to vary for more than an order of magnitude. Our results suggest that surface topology is a crucial consideration for the development of medical implants or biosensors whose performances are strongly influenced by the cell-to-material interface. We anticipate that the method can be used to explore the direct interaction of cells/tissue with medical devices such as metal implants in the future. PMID:28682058

Ballistic impact response of lipid membranes.

PubMed

Zhang, Yao; Meng, Zhaoxu; Qin, Xin; Keten, Sinan

2018-03-08

Therapeutic agent loaded micro and nanoscale particles as high-velocity projectiles can penetrate cells and tissues, thereby serving as gene and drug delivery vehicles for direct and rapid internalization. Despite recent progress in developing micro/nanoscale ballistic tools, the underlying biophysics of how fast projectiles deform and penetrate cell membranes is still poorly understood. To understand the rate and size-dependent penetration processes, we present coarse-grained molecular dynamics simulations of the ballistic impact of spherical projectiles on lipid membranes. Our simulations reveal that upon impact, the projectile can pursue one of three distinct pathways. At low velocities below the critical penetration velocity, projectiles rebound off the surface. At intermediate velocities, penetration occurs after the projectile deforms the membrane into a tubular thread. At very high velocities, rapid penetration occurs through localized membrane deformation without tubulation. Membrane tension, projectile velocity and size govern which phenomenon occurs, owing to their positive correlation with the reaction force generated between the projectile and the membrane during impact. Two critical membrane tension values dictate the boundaries among the three pathways for a given system, due to the rate dependence of the stress generated in the membrane. Our findings provide broad physical insights into the ballistic impact response of soft viscous membranes and guide design strategies for drug delivery through lipid membranes using micro/nanoscale ballistic tools.

The N-Terminal Amphipathic Helix of the Topological Specificity Factor MinE Is Associated with Shaping Membrane Curvature

PubMed Central

Shih, Yu-Ling; Huang, Kai-Fa; Lai, Hsin-Mei; Liao, Jiahn-Haur; Lee, Chai-Siah; Chang, Chiao-Min; Mak, Huey-Ming; Hsieh, Cheng-Wei; Lin, Chu-Chi

2011-01-01

Pole-to-pole oscillations of the Min proteins in Escherichia coli are required for the proper placement of the division septum. Direct interaction of MinE with the cell membrane is critical for the dynamic behavior of the Min system. In vitro, this MinE-membrane interaction led to membrane deformation; however, the underlying mechanism remained unclear. Here we report that MinE-induced membrane deformation involves the formation of an amphipathic helix of MinE2–9, which, together with the adjacent basic residues, function as membrane anchors. Biochemical evidence suggested that the membrane association induces formation of the helix, with the helical face, consisting of A2, L3, and F6, inserted into the membrane. Insertion of this helix into the cell membrane can influence local membrane curvature and lead to drastic changes in membrane topology. Accordingly, MinE showed characteristic features of protein-induced membrane tubulation and lipid clustering in in vitro reconstituted systems. In conclusion, MinE shares common protein signatures with a group of membrane trafficking proteins in eukaryotic cells. These MinE signatures appear to affect membrane curvature. PMID:21738659

Real-time detection of antibiotic activity by measuring nanometer-scale bacterial deformation

NASA Astrophysics Data System (ADS)

Iriya, Rafael; Syal, Karan; Jing, Wenwen; Mo, Manni; Yu, Hui; Haydel, Shelley E.; Wang, Shaopeng; Tao, Nongjian

2017-12-01

Diagnosing antibiotic-resistant bacteria currently requires sensitive detection of phenotypic changes associated with antibiotic action on bacteria. Here, we present an optical imaging-based approach to quantify bacterial membrane deformation as a phenotypic feature in real-time with a nanometer scale (˜9 nm) detection limit. Using this approach, we found two types of antibiotic-induced membrane deformations in different bacterial strains: polymyxin B induced relatively uniform spatial deformation of Escherichia coli O157:H7 cells leading to change in cellular volume and ampicillin-induced localized spatial deformation leading to the formation of bulges or protrusions on uropathogenic E. coli CFT073 cells. We anticipate that the approach will contribute to understanding of antibiotic phenotypic effects on bacteria with a potential for applications in rapid antibiotic susceptibility testing.

Elastic behavior of a red blood cell with the membrane's nonuniform natural state: equilibrium shape, motion transition under shear flow, and elongation during tank-treading motion.

PubMed

Tsubota, Ken-Ichi; Wada, Shigeo; Liu, Hao

2014-08-01

Direct numerical simulations of the mechanics of a single red blood cell (RBC) were performed by considering the nonuniform natural state of the elastic membrane. A RBC was modeled as an incompressible viscous fluid encapsulated by an elastic membrane. The in-plane shear and area dilatation deformations of the membrane were modeled by Skalak constitutive equation, while out-of-plane bending deformation was formulated by the spring model. The natural state of the membrane with respect to in-plane shear deformation was modeled as a sphere ([Formula: see text]), biconcave disk shape ([Formula: see text]) and their intermediate shapes ([Formula: see text]) with the nonuniformity parameter [Formula: see text], while the natural state with respect to out-of-plane bending deformation was modeled as a flat plane. According to the numerical simulations, at an experimentally measured in-plane shear modulus of [Formula: see text] and an out-of-plane bending rigidity of [Formula: see text] of the cell membrane, the following results were obtained. (i) The RBC shape at equilibrium was biconcave discoid for [Formula: see text] and cupped otherwise; (ii) the experimentally measured fluid shear stress at the transition between tumbling and tank-treading motions under shear flow was reproduced for [Formula: see text]; (iii) the elongation deformation of the RBC during tank-treading motion from the simulation was consistent with that from in vitro experiments, irrespective of the [Formula: see text] value. Based on our RBC modeling, the three phenomena (i), (ii), and (iii) were mechanically consistent for [Formula: see text]. The condition [Formula: see text] precludes a biconcave discoid shape at equilibrium (i); however, it gives appropriate fluid shear stress at the motion transition under shear flow (ii), suggesting that a combined effect of [Formula: see text] and the natural state with respect to out-of-plane bending deformation is necessary for understanding details of the RBC mechanics at equilibrium. Our numerical results demonstrate that moderate nonuniformity in a membrane's natural state with respect to in-plane shear deformation plays a key role in RBC mechanics.

Models of dynamic extraction of lipid tethers from cell membranes.

PubMed

Nowak, Sarah A; Chou, Tom

2010-05-07

When a ligand that is bound to an integral membrane receptor is pulled, the membrane and the underlying cytoskeleton can deform before either the membrane delaminates from the cytoskeleton or the ligand detaches from the receptor. If the membrane delaminates from the cytoskeleton, it may be further extruded and form a membrane tether. We develop a phenomenological model for this process by assuming that deformations obey Hooke's law up to a critical force at which the cell membrane locally detaches from the cytoskeleton and a membrane tether forms. We compute the probability of tether formation and show that tethers can be extruded only within an intermediate range of force loading rates and pulling velocities. The mean tether length that arises at the moment of ligand detachment is computed as are the force loading rates and pulling velocities that yield the longest tethers.

Shock Wave-Induced Damage and Poration in Eukaryotic Cell Membranes.

PubMed

López-Marín, Luz M; Millán-Chiu, Blanca E; Castaño-González, Karen; Aceves, Carmen; Fernández, Francisco; Varela-Echavarría, Alfredo; Loske, Achim M

2017-02-01

Shock waves are known to permeabilize eukaryotic cell membranes, which may be a powerful tool for a variety of drug delivery applications. However, the mechanisms involved in shock wave-mediated membrane permeabilization are still poorly understood. In this study, the effects on both the permeability and the ultrastructural features of two human cell lineages were investigated after the application of underwater shock waves in vitro. Scanning Electron Microscopy of cells derived from a human embryo kidney (HEK)-293 and Michigan Cancer Foundation (MCF)-7 cells, an immortalized culture derived from human breast adenocarcinoma, showed a small amount of microvilli (as compared to control cells), the presence of hole-like structures, and a decrease in cell size after shock wave exposure. Interestingly, these effects were accompanied by the permeabilization of acid and macromolecular dyes and gene transfection. Trypan blue exclusion assays indicated that cell membranes were porated during shock wave treatment but resealed after a few seconds. Deformations of the cell membrane lasted for at least 5 min, allowing their observation in fixed cells. For each cell line, different shock wave parameters were needed to achieve cell membrane poration. This difference was correlated to successful gene transfection by shock waves. Our results demonstrate, for the first time, that shock waves induce transient micro- and submicrosized deformations at the cell membrane, leading to cell transfection and cell survival. They also indicate that ultrastructural analyses of cell surfaces may constitute a useful way to match the use of shock waves to different cells and settings.

A microfabricated platform with hydrogel arrays for 3D mechanical stimulation of cells.

PubMed

Liu, Haijiao; Usprech, Jenna; Sun, Yu; Simmons, Craig A

2016-04-01

Cellular microenvironments present cells with multiple stimuli, including not only soluble biochemical and insoluble matrix cues but also mechanical factors. Biomaterial array platforms have been used to combinatorially and efficiently probe and define two-dimensional (2D) and 3D microenvironmental cues to guide cell functions for tissue engineering applications. However, there are few examples of array platforms that include dynamic mechanical forces, particularly to enable stretching of 3D cell-seeded biomaterials, which is relevant to engineering connective and cardiovascular tissues. Here we present a deformable membrane platform that enables 3D dynamic mechanical stretch of arrayed biomaterial constructs. Cell-seeded polyethylene glycol norbornene (PEG-NB) hydrogels were bound to miniaturized deformable membranes via a thiol-ene reaction with off-stoichiometry thiol-ene based polydimethylsiloxane (OSTE-PDMS) as the membrane material. Bonding to OSTE-PDMS enabled the 3D hydrogel microconstructs to be cyclically deformed and stretched by the membrane. As a first demonstration, human mesenchymal stromal cells (MSCs) embedded in PEG-NB were stretched for several days. They were found to be viable, spread in the 3D hydrogels, and exhibited a contractile myofibroblast phenotype when exposed to dynamic 3D mechanical deformation. This platform, which is readily scalable to larger arrays, enables systematic interrogation of the relationships between combinations of 3D mechanobiological cues and cellular responses, and thus has the potential to identify strategies to predictably control the construction of functional engineered tissues. Current high-throughput biomaterial screening approaches fail to consider the effects of dynamic mechanical stimulation, despite its importance in a wide variety of regenerative medicine applications. To meet this need, we developed a deformable membrane platform that enables 3D dynamic stretch of arrayed biomaterial constructs. Our approach combines microtechnologies fabricated with off-stoichiometry thiol-ene based polydimethylsiloxane membranes that can covalently bond cell-seeded polyethylene glycol norbornene 3D hydrogels, a model biomaterial with tunable adhesive, elastic and degradation characteristics. As a first demonstration, we show that human mesenchymal stromal cells embedded in hydrogels and subjected to dynamic mechanical stimulation undergo myofibroblast differentiation. This system is readily scaled up to larger arrays, and will enable systematic and efficient screening of combinations of 3D mechanobiological and biomaterial cues on cell fate and function. Copyright © 2015 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Direct Cytoskeleton Forces Cause Membrane Softening in Red Blood Cells

PubMed Central

Rodríguez-García, Ruddi; López-Montero, Iván; Mell, Michael; Egea, Gustavo; Gov, Nir S.; Monroy, Francisco

2015-01-01

Erythrocytes are flexible cells specialized in the systemic transport of oxygen in vertebrates. This physiological function is connected to their outstanding ability to deform in passing through narrow capillaries. In recent years, there has been an influx of experimental evidence of enhanced cell-shape fluctuations related to metabolically driven activity of the erythroid membrane skeleton. However, no direct observation of the active cytoskeleton forces has yet been reported to our knowledge. Here, we show experimental evidence of the presence of temporally correlated forces superposed over the thermal fluctuations of the erythrocyte membrane. These forces are ATP-dependent and drive enhanced flickering motions in human erythrocytes. Theoretical analyses provide support for a direct force exerted on the membrane by the cytoskeleton nodes as pulses of well-defined average duration. In addition, such metabolically regulated active forces cause global membrane softening, a mechanical attribute related to the functional erythroid deformability. PMID:26083919

Kinematics of red cell aspiration by fluorescence-imaged microdeformation.

PubMed

Discher, D E; Mohandas, N

1996-10-01

Maps of fluorescing red cell membrane components on a pipette-aspirated projection are quantitated in an effort to elucidate and unify the heterogeneous kinematics of deformation. Transient gradients of diffusing fluorescent lipid first demonstrate the fluidity of an otherwise uniform-density bilayer and corroborate a "universal" calibration scale for relative surface density. A steep but smooth and stable gradient in the densities of the skeleton components spectrin, actin, and protein 4.1 is used to estimate large elastic strains along the aspirated skeleton. The deformation fields are argued to be an unhindered response to loading in the surface normal direction. Density maps intermediate to those of the compressible skeleton and fluid bilayer are exhibited by particular transmembrane proteins (e.g., Band 3) and yield estimates for the skeleton-connected fractions. Such connected proteins appear to occupy a significant proportion of the undeformed membrane surface and can lead to steric exclusion of unconnected integral membrane proteins from regions of network condensation. Consistent with membrane repatterning kinematics in reversible deformation, final vesiculation of the projection tip produces a cell fragment concentrated in freely diffusing proteins but depleted of skeleton.

Hydroxycarbamide-Induced Changes in E/beta Thalassemia Red Blood Cells

PubMed Central

Sylvia, Singer T.; Elliott, Vichinsky; Sandra, Larkin; Nancy, Olivieri; Nancy, Sweeters; Frans, Kuypers A.

2010-01-01

In thalassemia, fetal hemoglobin (HbF) augmentation with hydroxycarbamide (also known as hydroxyurea) is not always successful. The expected parallel effects on RBC membrane deformability, cell hydration and membrane phospholipid organization, all important for extending RBC life span and increasing Hb, have been infrequently examined. We analyzed these characteristics in 15 non-transfused E/β 0 thalassemia patients treated with HU (mean 10.2 months). Membrane deformability and cell hydration mildly improved in association with increased HbF levels approaching statistical significance (r = 0.51, p=0.06). All measures improved considerably splenectomized patients. These findings underscore the disappointing results of hydroxyurea treatment in clinical trials; and the importance of examining the effect on red cell characteristics for the development and understanding of HbF-enhancing agents. PMID:18821710

A numerical method for osmotic water flow and solute diffusion with deformable membrane boundaries in two spatial dimension

NASA Astrophysics Data System (ADS)

Yao, Lingxing; Mori, Yoichiro

2017-12-01

Osmotic forces and solute diffusion are increasingly seen as playing a fundamental role in cell movement. Here, we present a numerical method that allows for studying the interplay between diffusive, osmotic and mechanical effects. An osmotically active solute obeys a advection-diffusion equation in a region demarcated by a deformable membrane. The interfacial membrane allows transmembrane water flow which is determined by osmotic and mechanical pressure differences across the membrane. The numerical method is based on an immersed boundary method for fluid-structure interaction and a Cartesian grid embedded boundary method for the solute. We demonstrate our numerical algorithm with the test case of an osmotic engine, a recently proposed mechanism for cell propulsion.

The dynamic behavior of chemically "stiffened" red blood cells in microchannel flows.

PubMed

Forsyth, Alison M; Wan, Jiandi; Ristenpart, William D; Stone, Howard A

2010-07-01

The rigidity of red blood cells (RBCs) plays an important role in whole blood viscosity and is correlated with several cardiovascular diseases. Two chemical agents that are commonly used to study cell deformation are diamide and glutaraldehyde. Despite diamide's common usage, there are discrepancies in the literature surrounding diamide's effect on the deformation of RBCs in shear and pressure-driven flows; in particular, shear flow experiments have shown that diamide stiffens cells, while pressure-driven flow in capillaries did not give this result. We performed pressure-driven flow experiments with RBCs in a microfluidic constriction and quantified the cell dynamics using high-speed imaging. Diamide, which affects RBCs by cross-linking spectrin skeletal membrane proteins, did not reduce deformation and showed an unchanged effective strain rate when compared to healthy cells. In contrast, glutaraldehyde, which is a non-specific fixative that acts on all components of the cell, did reduce deformation and showed increased instances of tumbling, both of which are characteristic features of stiffened, or rigidified, cells. Because glutaraldehyde increases the effective viscosity of the cytoplasm and lipid membrane while diamide does not, one possible explanation for our results is that viscous effects in the cytoplasm and/or lipid membrane are a dominant factor in dictating dynamic responses of RBCs in pressure-driven flows. Finally, literature on the use of diamide as a stiffening agent is summarized, and provides supporting evidence for our conclusions. Copyright 2010 Elsevier Inc. All rights reserved.

Selective flow-induced vesicle rupture to sort by membrane mechanical properties

NASA Astrophysics Data System (ADS)

Pommella, Angelo; Brooks, Nicholas J.; Seddon, John M.; Garbin, Valeria

2015-08-01

Vesicle and cell rupture caused by large viscous stresses in ultrasonication is central to biomedical and bioprocessing applications. The flow-induced opening of lipid membranes can be exploited to deliver drugs into cells, or to recover products from cells, provided that it can be obtained in a controlled fashion. Here we demonstrate that differences in lipid membrane and vesicle properties can enable selective flow-induced vesicle break-up. We obtained vesicle populations with different membrane properties by using different lipids (SOPC, DOPC, or POPC) and lipid:cholesterol mixtures (SOPC:chol and DOPC:chol). We subjected vesicles to large deformations in the acoustic microstreaming flow generated by ultrasound-driven microbubbles. By simultaneously deforming vesicles with different properties in the same flow, we determined the conditions in which rupture is selective with respect to the membrane stretching elasticity. We also investigated the effect of vesicle radius and excess area on the threshold for rupture, and identified conditions for robust selectivity based solely on the mechanical properties of the membrane. Our work should enable new sorting mechanisms based on the difference in membrane composition and mechanical properties between different vesicles, capsules, or cells.

Selective flow-induced vesicle rupture to sort by membrane mechanical properties

PubMed Central

Pommella, Angelo; Brooks, Nicholas J.; Seddon, John M.; Garbin, Valeria

2015-01-01

Vesicle and cell rupture caused by large viscous stresses in ultrasonication is central to biomedical and bioprocessing applications. The flow-induced opening of lipid membranes can be exploited to deliver drugs into cells, or to recover products from cells, provided that it can be obtained in a controlled fashion. Here we demonstrate that differences in lipid membrane and vesicle properties can enable selective flow-induced vesicle break-up. We obtained vesicle populations with different membrane properties by using different lipids (SOPC, DOPC, or POPC) and lipid:cholesterol mixtures (SOPC:chol and DOPC:chol). We subjected vesicles to large deformations in the acoustic microstreaming flow generated by ultrasound-driven microbubbles. By simultaneously deforming vesicles with different properties in the same flow, we determined the conditions in which rupture is selective with respect to the membrane stretching elasticity. We also investigated the effect of vesicle radius and excess area on the threshold for rupture, and identified conditions for robust selectivity based solely on the mechanical properties of the membrane. Our work should enable new sorting mechanisms based on the difference in membrane composition and mechanical properties between different vesicles, capsules, or cells. PMID:26302783

A multiscale red blood cell model with accurate mechanics, rheology, and dynamics.

PubMed

Fedosov, Dmitry A; Caswell, Bruce; Karniadakis, George Em

2010-05-19

Red blood cells (RBCs) have highly deformable viscoelastic membranes exhibiting complex rheological response and rich hydrodynamic behavior governed by special elastic and bending properties and by the external/internal fluid and membrane viscosities. We present a multiscale RBC model that is able to predict RBC mechanics, rheology, and dynamics in agreement with experiments. Based on an analytic theory, the modeled membrane properties can be uniquely related to the experimentally established RBC macroscopic properties without any adjustment of parameters. The RBC linear and nonlinear elastic deformations match those obtained in optical-tweezers experiments. The rheological properties of the membrane are compared with those obtained in optical magnetic twisting cytometry, membrane thermal fluctuations, and creep followed by cell recovery. The dynamics of RBCs in shear and Poiseuille flows is tested against experiments and theoretical predictions, and the applicability of the latter is discussed. Our findings clearly indicate that a purely elastic model for the membrane cannot accurately represent the RBC's rheological properties and its dynamics, and therefore accurate modeling of a viscoelastic membrane is necessary. Copyright 2010 Biophysical Society. Published by Elsevier Inc. All rights reserved.

A Multiscale Red Blood Cell Model with Accurate Mechanics, Rheology, and Dynamics

PubMed Central

Fedosov, Dmitry A.; Caswell, Bruce; Karniadakis, George Em

2010-01-01

Abstract Red blood cells (RBCs) have highly deformable viscoelastic membranes exhibiting complex rheological response and rich hydrodynamic behavior governed by special elastic and bending properties and by the external/internal fluid and membrane viscosities. We present a multiscale RBC model that is able to predict RBC mechanics, rheology, and dynamics in agreement with experiments. Based on an analytic theory, the modeled membrane properties can be uniquely related to the experimentally established RBC macroscopic properties without any adjustment of parameters. The RBC linear and nonlinear elastic deformations match those obtained in optical-tweezers experiments. The rheological properties of the membrane are compared with those obtained in optical magnetic twisting cytometry, membrane thermal fluctuations, and creep followed by cell recovery. The dynamics of RBCs in shear and Poiseuille flows is tested against experiments and theoretical predictions, and the applicability of the latter is discussed. Our findings clearly indicate that a purely elastic model for the membrane cannot accurately represent the RBC's rheological properties and its dynamics, and therefore accurate modeling of a viscoelastic membrane is necessary. PMID:20483330

Membrane Deformation and Permeabilization Caused by Microplasma Irradiation

NASA Astrophysics Data System (ADS)

Motomura, Hideki; Nagaiwa, Hidenori; Yamamoto, Kenta; Kido, Yugo; Ikeda, Yoshihisa; Satoh, Susumu; Jinno, Masafumi

2016-09-01

The microplasma irradiation achieves high gene taransfection efficiency and high cell survivability simultaneously. For this purpose, we have developed a special plasma source using a microcapillary electrode. However, it is not clear how the stimuli of effective factors generated by plasma, such as current, charge, field, chemical species, cause transfection. In this study, we used artificial cell which is a spherical vesicle consisting of a lipid bilayer to visualize membrane dynamics and permeabilization caused by microplasma irradiation. Dioleoyl phosphatidylcholine (DOPC) was used as phospholipid molecules forming the lipid bilayer. The artificial cells were prepared by natural swelling method. Fluorescent labeled polyethylene glycol (PEG) polymers (Nanocs, MPEG Fluorescein, MW = 1000) were encapsulated in the artificial cells. The artificial cells were exposed to the microplasma for 5 ms and 10-20% of decrease of the dye fluorescence in the artificial cells was observed. This result suggests the outflow of the MPEG polymers through temporary poration or deformation of the lipid bilayer. The membrane deformation dynamics was directly observed with a microscope and the relationship to the polymer outflow will be shown at the conference. This work was partly supported by a Grant-in-Aid (25108509 and 15H00896) from JSPS and a grant from Ehime University.

A study of the dynamic properties of the human red blood cell membrane using quasi-elastic light-scattering spectroscopy.

PubMed

Tishler, R B; Carlson, F D

1993-12-01

A quasi-elastic light-scattering (QELS) microscope spectrometer was used to study the dynamic properties of the membrane/cytoskeleton of individual human red blood cells (RBCs). QELS is a spectroscopic technique that measures intensity fluctuations of laser light scattered from a sample. The intensity fluctuations were analyzed using power spectra and the intensity autocorrelation function, g(2)(tau), which was approximated with a single exponential. The value of the correlation time, Tcorr, was used for comparing results. Motion of the RBC membrane/cytoskeleton was previously identified as the source of the QELS signal from the RBC (R. B. Tishler and F. D. Carlson, 1987. Biophys. J. 51:993-997), and additional data supporting that conclusion are presented. Similar results were obtained from anucleate mammalian RBCs that have structures similar to that of the human RBC, but not for morphologically distinct, nucleated RBCs. The effect of altering the physical properties of the cytoplasm and the membrane/cytoskeleton was also studied. Osmotically increasing the cytoplasmic viscosity led to significant increases in Tcorr. Increasing the membrane cholesterol content and increasing the intracellular calcium content both led to decreased deformability of the human RBC. In both cases, the modified cells with decreased deformability showed an increase in Tcorr, demonstrating that QELS could measure biochemically induced changes of the membrane/cytoskeleton. Physiological changes were measured in studies of age-separated RBC populations which showed that Tcorr was increased in the older, less deformable cells.

Indentation of Graphene-Covered Atomic Force Microscopy Probe Across a Lipid Bilayer Membrane: Effect of Tip Shape, Size, and Surface Hydrophobicity.

PubMed

Lv, Kang; Li, Yinfeng

2018-06-21

Understanding the interaction of graphene with cell membranes is crucial to the development of graphene-based biological applications and the management of graphene safety issues. To help reveal the key factors controlling the interaction between graphene and cell membranes, here we adopt the dissipative particle dynamics method to analyze the evolution of interaction force and free energy as the graphene-covered atomic force microscopy (AFM) probe indents across a lipid bilayer. The simulation results show that the graphene-covered AFM probe can cause severe deformation of the cell membrane which drives the lipid molecule to adsorb and diffuse at the surface of graphene. The breakthrough force and free energy are calculated to study the effects of the tip shape, size, and surface hydrophobicity on the piercing behaviors of graphene-covered AFM. In addition, the deformation of cell membrane can decrease the dependency of the breakthrough force on the tip shape. The analysis of surface functionalization suggests that the horizontal patterns on graphene can change the preferred orientation in the penetration process, but the vertical patterns on graphene may disrupt the cell membrane. What's more, the bending stiffness of graphene has little influence on the penetration process as graphene pierces into the cell membrane. These results provide useful guidelines for the molecular design of graphene materials with controllable cell penetrability.

Real-time detection of antibiotic activity by measuring nanometer-scale bacterial deformation.

PubMed

Iriya, Rafael; Syal, Karan; Jing, Wenwen; Mo, Manni; Yu, Hui; Haydel, Shelley E; Wang, Shaopeng; Tao, Nongjian

2017-12-01

Diagnosing antibiotic-resistant bacteria currently requires sensitive detection of phenotypic changes associated with antibiotic action on bacteria. Here, we present an optical imaging-based approach to quantify bacterial membrane deformation as a phenotypic feature in real-time with a nanometer scale (∼9  nm) detection limit. Using this approach, we found two types of antibiotic-induced membrane deformations in different bacterial strains: polymyxin B induced relatively uniform spatial deformation of Escherichia coli O157:H7 cells leading to change in cellular volume and ampicillin-induced localized spatial deformation leading to the formation of bulges or protrusions on uropathogenic E. coli CFT073 cells. We anticipate that the approach will contribute to understanding of antibiotic phenotypic effects on bacteria with a potential for applications in rapid antibiotic susceptibility testing. (2017) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE).

Salt Bridge Formation between the I-BAR Domain and Lipids Increases Lipid Density and Membrane Curvature.

PubMed

Takemura, Kazuhiro; Hanawa-Suetsugu, Kyoko; Suetsugu, Shiro; Kitao, Akio

2017-07-28

The BAR domain superfamily proteins sense or induce curvature in membranes. The inverse-BAR domain (I-BAR) is a BAR domain that forms a straight "zeppelin-shaped" dimer. The mechanisms by which IRSp53 I-BAR binds to and deforms a lipid membrane are investigated here by all-atom molecular dynamics simulation (MD), binding energy analysis, and the effects of mutation experiments on filopodia on HeLa cells. I-BAR adopts a curved structure when crystallized, but adopts a flatter shape in MD. The binding of I-BAR to membrane was stabilized by ~30 salt bridges, consistent with experiments showing that point mutations of the interface residues have little effect on the binding affinity whereas multiple mutations have considerable effect. Salt bridge formation increases the local density of lipids and deforms the membrane into a concave shape. In addition, the point mutations that break key intra-molecular salt bridges within I-BAR reduce the binding affinity; this was confirmed by expressing these mutants in HeLa cells and observing their effects. The results indicate that the stiffness of I-BAR is important for membrane deformation, although I-BAR does not act as a completely rigid template.

A zebrafish embryo behaves both as a "cortical shell-liquid core" structure and a homogeneous solid when experiencing mechanical forces.

PubMed

Liu, Fei; Wu, Dan; Chen, Ken

2014-12-01

Mechanical properties are vital for living cells, and various models have been developed to study the mechanical behavior of cells. However, there is debate regarding whether a cell behaves more similarly to a "cortical shell-liquid core" structure (membrane-like) or a homogeneous solid (cytoskeleton-like) when experiencing stress by mechanical forces. Unlike most experimental methods, which concern the small-strain deformation of a cell, we focused on the mechanical behavior of a cell undergoing small to large strain by conducting microinjection experiments on zebrafish embryo cells. The power law with order of 1.5 between the injection force and the injection distance indicates that the cell behaves as a homogenous solid at small-strain deformation. The linear relation between the rupture force and the microinjector radius suggests that the embryo behaves as membrane-like when subjected to large-strain deformation. We also discuss the possible reasons causing the debate by analyzing the mechanical properties of F-actin filaments.

Role of the membrane cortex in neutrophil deformation in small pipets.

PubMed Central

Zhelev, D V; Needham, D; Hochmuth, R M

1994-01-01

The simplest model for a neutrophil in its "passive" state views the cell as consisting of a liquid-like cytoplasmic region surrounded by a membrane. The cell surface is in a state of isotropic contraction, which causes the cell to assume a spherical shape. This contraction is characterized by the cortical tension. The cortical tension shows a weak area dilation dependence, and it determines the elastic properties of the cell for small curvature deformations. At high curvature deformations in small pipets (with internal radii less than 1 micron), the measured critical suction pressure for cell flow into the pipet is larger than its estimate from the law of Laplace. A model is proposed where the region consisting of the cytoplasm membrane and the underlying cortex (having a finite thickness) is introduced at the cell surface. The mechanical properties of this region are characterized by the apparent cortical tension (defined as a free contraction energy per unit area) and the apparent bending modulus (introduced as a bending free energy per unit area) of its middle plane. The model predicts that for small curvature deformations (in pipets having radii larger than 1.2 microns) the role of the cortical thickness and the resistance for bending of the membrane-cortex complex is negligible. For high curvature deformations, they lead to elevated suction pressures above the values predicted from the law of Laplace. The existence of elevated suction pressures for pipets with radii from 1 micron down to 0.24 micron is found experimentally. The measured excess suction pressures cannot be explained only by the modified law of Laplace (for a cortex with finite thickness and negligible bending resistance), because it predicts unacceptable high cortical thicknesses (from 0.3 to 0.7 micron). It is concluded that the membrane-cortex complex has an apparent bending modulus from 1 x 10(-18) to 2 x 10(-18) J for a cortex with a thickness from 0.1 micron down to values much smaller than the radius of the smallest pipet (0.24 micron) used in this study. Images FIGURE 1 PMID:7948682

Red blood cell-deformability measurement: review of techniques.

PubMed

Musielak, M

2009-01-01

Cell-deformability characterization involves general measurement of highly complex relationships between cell biology and physical forces to which the cell is subjected. The review takes account of the modern technical solutions simulating the action of the force applied to the red blood cell in macro- and microcirculation. Diffraction ektacytometers and rheoscopes measure the mean deformability value for the total red blood cell population investigated and the deformation distribution index of individual cells, respectively. Deformation assays of a whole single cell are possible by means of optical tweezers. The single cell-measuring setups for micropipette aspiration and atomic force microscopy allow conducting a selective investigation of deformation parameters (e.g., cytoplasm viscosity, viscoelastic membrane properties). The distinction between instrument sensitivity to various RBC-rheological features as well as the influence of temperature on measurement are discussed. The reports quoted confront fascinating possibilities of the techniques with their medical applications since the RBC-deformability has the key position in the etiology of a wide range of conditions.

Microfluidic assay of the deformability of primitive erythroblasts.

PubMed

Zhou, Sitong; Huang, Yu-Shan; Kingsley, Paul D; Cyr, Kathryn H; Palis, James; Wan, Jiandi

2017-09-01

Primitive erythroblasts (precursors of red blood cells) enter vascular circulation during the embryonic period and mature while circulating. As a result, primitive erythroblasts constantly experience significant hemodynamic shear stress. Shear-induced deformation of primitive erythroblasts however, is poorly studied. In this work, we examined the deformability of primitive erythroblasts at physiologically relevant flow conditions in microfluidic channels and identified the regulatory roles of the maturation stage of primitive erythroblasts and cytoskeletal protein 4.1 R in shear-induced cell deformation. The results showed that the maturation stage affected the deformability of primitive erythroblasts significantly and that primitive erythroblasts at later maturational stages exhibited a better deformability due to a matured cytoskeletal structure in the cell membrane.

Recent progress on lipid lateral heterogeneity in plasma membranes: from rafts to submicrometric domains

PubMed Central

Carquin, Mélanie; D'Auria, Ludovic; Pollet, Hélène; Bongarzone, Ernesto R.; Tyteca, Donatienne

2016-01-01

The concept of transient nanometric domains known as lipid rafts has brought interest to reassess the validity of the Singer-Nicholson model of a fluid bilayer for cell membranes. However, this new view is still insufficient to explain the cellular control of surface lipid diversity or membrane deformability. During the past decade, the hypothesis that some lipids form large (submicrometric/mesoscale vs nanometric rafts) and stable (> min vs sec) membrane domains has emerged, largely based on indirect methods. Morphological evidence for stable submicrometric lipid domains, well-accepted for artificial and highly specialized biological membranes, was further reported for a variety of living cells from prokaryotes to yeast and mammalian cells. However, results remained questioned based on limitations of available fluorescent tools, use of poor lipid fixatives, and imaging artifacts due to non-resolved membrane projections. In this review, we will discuss recent evidence generated using powerful and innovative approaches such as lipid-specific toxin fragments that support the existence of submicrometric domains. We will integrate documented mechanisms involved in the formation and maintenance of these domains, and provide a perspective on their relevance on membrane deformability and regulation of membrane protein distribution. PMID:26738447

Vertical nanopillars for in situ probing of nuclear mechanics in adherent cells

PubMed Central

Hanson, Lindsey; Zhao, Wenting; Lou, Hsin-Ya; Lin, Ziliang Carter; Lee, Seok Woo; Chowdary, Praveen; Cui, Yi; Cui, Bianxiao

2016-01-01

The mechanical stability and deformability of the cell nucleus are crucial to many biological processes, including migration, proliferation and polarization. In vivo, the cell nucleus is frequently subjected to deformation on a variety of length and time scales, but current techniques for studying nuclear mechanics do not provide access to subnuclear deformation in live functioning cells. Here we introduce arrays of vertical nanopillars as a new method for the in situ study of nuclear deformability and the mechanical coupling between the cell membrane and the nucleus in live cells. Our measurements show that nanopillar-induced nuclear deformation is determined by nuclear stiffness, as well as opposing effects from actin and intermediate filaments. Furthermore, the depth, width and curvature of nuclear deformation can be controlled by varying the geometry of the nanopillar array. Overall, vertical nanopillar arrays constitute a novel approach for non-invasive, subcellular perturbation of nuclear mechanics and mechanotransduction in live cells. PMID:25984833

Dynamics of shear-induced ATP release from red blood cells.

PubMed

Wan, Jiandi; Ristenpart, William D; Stone, Howard A

2008-10-28

Adenosine triphosphate (ATP) is a regulatory molecule for many cell functions, both for intracellular and, perhaps less well known, extracellular functions. An important example of the latter involves red blood cells (RBCs), which help regulate blood pressure by releasing ATP as a vasodilatory signaling molecule in response to the increased shear stress inside arterial constrictions. Although shear-induced ATP release has been observed widely and is believed to be triggered by deformation of the cell membrane, the underlying mechanosensing mechanism inside RBCs is still controversial. Here, we use an in vitro microfluidic approach to investigate the dynamics of shear-induced ATP release from human RBCs with millisecond resolution. We demonstrate that there is a sizable delay time between the onset of increased shear stress and the release of ATP. This response time decreases with shear stress, but surprisingly does not depend significantly on membrane rigidity. Furthermore, we show that even though the RBCs deform significantly in short constrictions (duration of increased stress <3 ms), no measurable ATP is released. This critical timescale is commensurate with a characteristic membrane relaxation time determined from observations of the cell deformation by using high-speed video. Taken together our results suggest a model wherein the retraction of the spectrin-actin cytoskeleton network triggers the mechanosensitive ATP release and a shear-dependent membrane viscosity controls the rate of release.

Assembly of MreB filaments on liposome membranes: a synthetic biology approach.

PubMed

Maeda, Yusuke T; Nakadai, Tomoyoshi; Shin, Jonghyeon; Uryu, Kunihiro; Noireaux, Vincent; Libchaber, Albert

2012-02-17

The physical interaction between the cytoskeleton and the cell membrane is essential in defining the morphology of living organisms. In this study, we use a synthetic approach to polymerize bacterial MreB filaments inside phospholipid vesicles. When the proteins MreB and MreC are expressed inside the liposomes, the MreB cytoskeleton structure develops at the inner membrane. Furthermore, when purified MreB is used inside the liposomes, MreB filaments form a 4-10 μm rigid bundle structure and deform the lipid vesicles in physical contact with the vesicle inner membrane. These results indicate that the fibrillation of MreB filaments can take place either in close proximity of deformable lipid membrane or in the presence of associated protein. Our finding might be relevant for the self-assembly of cytoskeleton filaments toward the construction of artificial cell systems.

Membrane-mediated interaction between retroviral capsids

NASA Astrophysics Data System (ADS)

Zhang, Rui; Nguyen, Toan

2012-02-01

A retrovirus is an RNA virus that is replicated through a unique strategy of reverse transcription. Unlike regular enveloped viruses which are assembled inside the host cells, the assembly of retroviral capsids happens right on the cell membrane. During the assembly process, the partially formed capsids deform the membrane, giving rise to an elastic energy. When two such partial capsids approach each other, this elastic energy changes. Or in other words, the two partial capsids interact with each other via the membrane. This membrane mediated interaction between partial capsids plays an important role in the kinetics of the assembly process. In this work, this membrane mediated interaction is calculated both analytically and numerically. It is worth noting that the diferential equation determining the membrane shape in general nonlinear and cannot be solved analytically,except in the linear region of small deformations. And it is exactly the nonlinear regime that is important for the assembly kinetics of retroviruses as it provides a large energy barrier. The theory developed here is applicable to more generic cases of membrane mediated interactions between two membrane-embedded proteins.

Regulation of the actin cytoskeleton-plasma membrane interplay by phosphoinositides.

PubMed

Saarikangas, Juha; Zhao, Hongxia; Lappalainen, Pekka

2010-01-01

The plasma membrane and the underlying cortical actin cytoskeleton undergo continuous dynamic interplay that is responsible for many essential aspects of cell physiology. Polymerization of actin filaments against cellular membranes provides the force for a number of cellular processes such as migration, morphogenesis, and endocytosis. Plasma membrane phosphoinositides (especially phosphatidylinositol bis- and trisphosphates) play a central role in regulating the organization and dynamics of the actin cytoskeleton by acting as platforms for protein recruitment, by triggering signaling cascades, and by directly regulating the activities of actin-binding proteins. Furthermore, a number of actin-associated proteins, such as BAR domain proteins, are capable of directly deforming phosphoinositide-rich membranes to induce plasma membrane protrusions or invaginations. Recent studies have also provided evidence that the actin cytoskeleton-plasma membrane interactions are misregulated in a number of pathological conditions such as cancer and during pathogen invasion. Here, we summarize the wealth of knowledge on how the cortical actin cytoskeleton is regulated by phosphoinositides during various cell biological processes. We also discuss the mechanisms by which interplay between actin dynamics and certain membrane deforming proteins regulate the morphology of the plasma membrane.

C60 fullerene localization and membrane interactions in RAW 264.7 immortalized mouse macrophages

NASA Astrophysics Data System (ADS)

Russ, K. A.; Elvati, P.; Parsonage, T. L.; Dews, A.; Jarvis, J. A.; Ray, M.; Schneider, B.; Smith, P. J. S.; Williamson, P. T. F.; Violi, A.; Philbert, M. A.

2016-02-01

There continues to be a significant increase in the number and complexity of hydrophobic nanomaterials that are engineered for a variety of commercial purposes making human exposure a significant health concern. This study uses a combination of biophysical, biochemical and computational methods to probe potential mechanisms for uptake of C60 nanoparticles into various compartments of living immune cells. Cultures of RAW 264.7 immortalized murine macrophage were used as a canonical model of immune-competent cells that are likely to provide the first line of defense following inhalation. Modes of entry studied were endocytosis/pinocytosis and passive permeation of cellular membranes. The evidence suggests marginal uptake of C60 clusters is achieved through endocytosis/pinocytosis, and that passive diffusion into membranes provides a significant source of biologically-available nanomaterial. Computational modeling of both a single molecule and a small cluster of fullerenes predicts that low concentrations of fullerenes enter the membrane individually and produce limited perturbation; however, at higher concentrations the clusters in the membrane causes deformation of the membrane. These findings are bolstered by nuclear magnetic resonance (NMR) of model membranes that reveal deformation of the cell membrane upon exposure to high concentrations of fullerenes. The atomistic and NMR models fail to explain escape of the particle out of biological membranes, but are limited to idealized systems that do not completely recapitulate the complexity of cell membranes. The surprising contribution of passive modes of cellular entry provides new avenues for toxicological research that go beyond the pharmacological inhibition of bulk transport systems such as pinocytosis.There continues to be a significant increase in the number and complexity of hydrophobic nanomaterials that are engineered for a variety of commercial purposes making human exposure a significant health concern. This study uses a combination of biophysical, biochemical and computational methods to probe potential mechanisms for uptake of C60 nanoparticles into various compartments of living immune cells. Cultures of RAW 264.7 immortalized murine macrophage were used as a canonical model of immune-competent cells that are likely to provide the first line of defense following inhalation. Modes of entry studied were endocytosis/pinocytosis and passive permeation of cellular membranes. The evidence suggests marginal uptake of C60 clusters is achieved through endocytosis/pinocytosis, and that passive diffusion into membranes provides a significant source of biologically-available nanomaterial. Computational modeling of both a single molecule and a small cluster of fullerenes predicts that low concentrations of fullerenes enter the membrane individually and produce limited perturbation; however, at higher concentrations the clusters in the membrane causes deformation of the membrane. These findings are bolstered by nuclear magnetic resonance (NMR) of model membranes that reveal deformation of the cell membrane upon exposure to high concentrations of fullerenes. The atomistic and NMR models fail to explain escape of the particle out of biological membranes, but are limited to idealized systems that do not completely recapitulate the complexity of cell membranes. The surprising contribution of passive modes of cellular entry provides new avenues for toxicological research that go beyond the pharmacological inhibition of bulk transport systems such as pinocytosis. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr07003a

Red blood cell dynamics: from cell deformation to ATP release.

PubMed

Wan, Jiandi; Forsyth, Alison M; Stone, Howard A

2011-10-01

The mechanisms of red blood cell (RBC) deformation under both static and dynamic, i.e., flow, conditions have been studied extensively since the mid 1960s. Deformation-induced biochemical reactions and possible signaling in RBCs, however, were proposed only fifteen years ago. Therefore, the fundamental relationship between RBC deformation and cellular signaling dynamics i.e., mechanotransduction, remains incompletely understood. Quantitative understanding of the mechanotransductive pathways in RBCs requires integrative studies of physical models of RBC deformation and cellular biochemical reactions. In this article we review the physical models of RBC deformation, spanning from continuum membrane mechanics to cellular skeleton dynamics under both static and flow conditions, and elaborate the mechanistic links involved in deformation-induced ATP release. This journal is © The Royal Society of Chemistry 2011

Cell Membrane Deformation Induced by a Fibronectin-Coated Polystyrene Microbead in a 200-MHz Acoustic Trap

PubMed Central

Hwang, Jae Youn; Lee, Changyang; Lam, Kwok Ho; Kim, Hyung Ham; Lee, Jungwoo; Shung, K. Kirk

2014-01-01

The measurement of cell mechanics is crucial for a better understanding of cellular responses during the progression of certain diseases and for the identification of the cell’s nature. Many techniques using optical tweezers, atomic force microscopy, and micro-pipettes have been developed to probe and manipulate cells in the spatial domain. In particular, we recently proposed a two-dimensional acoustic trapping method as an alternative technique for small particle manipulation. Although the proposed method may have advantages over optical tweezers, its applications to cellular mechanics have not yet been vigorously investigated. This study represents an initial attempt to use acoustic tweezers as a tool in the field of cellular mechanics in which cancer cell membrane deformability is studied. A press-focused 193-MHz single-element lithium niobate (LiNbO3) transducer was designed and fabricated to trap a 5-µm polystyrene microbead near the ultrasound beam focus. The microbeads were coated with fibronectin, and trapped before being attached to the surface of a human breast cancer cell (MCF-7). The cell membrane was then stretched by remotely pulling a cell-attached microbead with the acoustic trap. The maximum cell membrane stretched lengths were measured to be 0.15, 0.54, and 1.41 µm at input voltages to the transducer of 6.3, 9.5, and 12.6 Vpp, respectively. The stretched length was found to increase nonlinearly as a function of the voltage input. No significant cytotoxicity was observed to result from the bead or the trapping force on the cell during or after the deformation procedure. Hence, the results convincingly demonstrated the possible application of the acoustic trapping technique as a tool for cell manipulation. PMID:24569245

Fixed charge in the cell membrane

PubMed Central

Elul, R.

1967-01-01

1. Focal electric field was generated by passing a current of 5 × 10-7 to 1 × 10-5 A from a micropipette into the culture medium. Movement of cells at a distance of 5-50 μ from the electrode tip was observed. In case of cells embedded in the culture only local deformation of the membrane was observed. 2. The cell species explored included neurones, glia, muscle fibres, connective cells, malignant cells and erythrocytes. All cells responded in a similar manner to the electric field, and the current required was in the same range. 3. Cells were attracted to a positive micropipette and repelled from a negative one: the only exception was observed in certain malignant cells which moved in the opposite direction. 4. Movement and membrane deformation could be obtained with electrodes filled with various concentrated and isotonic solutions. The composition of the culture medium also had no qualitative influence on these effects. 5. Metabolic poisons or rupture of the cell membrane had no effect on the movement. Isolated membrane fragments showed movement similar to that of intact cells. 6. The possibility of artifacts due to proximity of the focal electrode is considered. It is shown that electro-osmosis cannot account for the present observations. Some other artifacts are also excluded. 7. It is proposed that the most satisfactory way to account for the present observations is by a membrane carrying negative fixed charge of the order of 2·5 × 103 e.s.u./cm2. Some physiological consequences of presence of negative charge in the membrane are briefly discussed. ImagesFig. 1Fig. 2Fig. 3 PMID:6040152

Tunable Microfluidic Devices for Hydrodynamic Fractionation of Cells and Beads: A Review

PubMed Central

Alvankarian, Jafar; Majlis, Burhanuddin Yeop

2015-01-01

The adjustable microfluidic devices that have been developed for hydrodynamic-based fractionation of beads and cells are important for fast performance tunability through interaction of mechanical properties of particles in fluid flow and mechanically flexible microstructures. In this review, the research works reported on fabrication and testing of the tunable elastomeric microfluidic devices for applications such as separation, filtration, isolation, and trapping of single or bulk of microbeads or cells are discussed. Such microfluidic systems for rapid performance alteration are classified in two groups of bulk deformation of microdevices using external mechanical forces, and local deformation of microstructures using flexible membrane by pneumatic pressure. The main advantage of membrane-based tunable systems has been addressed to be the high capability of integration with other microdevice components. The stretchable devices based on bulk deformation of microstructures have in common advantage of simplicity in design and fabrication process. PMID:26610519

Kinematics of red cell aspiration by fluorescence-imaged microdeformation.

PubMed Central

Discher, D E; Mohandas, N

1996-01-01

Maps of fluorescing red cell membrane components on a pipette-aspirated projection are quantitated in an effort to elucidate and unify the heterogeneous kinematics of deformation. Transient gradients of diffusing fluorescent lipid first demonstrate the fluidity of an otherwise uniform-density bilayer and corroborate a "universal" calibration scale for relative surface density. A steep but smooth and stable gradient in the densities of the skeleton components spectrin, actin, and protein 4.1 is used to estimate large elastic strains along the aspirated skeleton. The deformation fields are argued to be an unhindered response to loading in the surface normal direction. Density maps intermediate to those of the compressible skeleton and fluid bilayer are exhibited by particular transmembrane proteins (e.g., Band 3) and yield estimates for the skeleton-connected fractions. Such connected proteins appear to occupy a significant proportion of the undeformed membrane surface and can lead to steric exclusion of unconnected integral membrane proteins from regions of network condensation. Consistent with membrane repatterning kinematics in reversible deformation, final vesiculation of the projection tip produces a cell fragment concentrated in freely diffusing proteins but depleted of skeleton. Images FIGURE 1 FIGURE 2 FIGURE 4 FIGURE 5 FIGURE 7 FIGURE 8 FIGURE 9 FIGURE 10 FIGURE 11 PMID:8889146

Cell membrane deformation and bioeffects produced by tandem bubble-induced jetting flow

PubMed Central

Yuan, Fang; Yang, Chen; Zhong, Pei

2015-01-01

Cavitation with bubble–bubble interaction is a fundamental feature in therapeutic ultrasound. However, the causal relationships between bubble dynamics, associated flow motion, cell deformation, and resultant bioeffects are not well elucidated. Here, we report an experimental system for tandem bubble (TB; maximum diameter = 50 ± 2 μm) generation, jet formation, and subsequent interaction with single HeLa cells patterned on fibronectin-coated islands (32 × 32 μm) in a microfluidic chip. We have demonstrated that pinpoint membrane poration can be produced at the leading edge of the HeLa cell in standoff distance Sd ≤ 30 μm, driven by the transient shear stress associated with TB-induced jetting flow. The cell membrane deformation associated with a maximum strain rate on the order of 104 s−1 was heterogeneous. The maximum area strain (εA,M) decreased exponentially with Sd (also influenced by adhesion pattern), a feature that allows us to create distinctly different treatment outcome (i.e., necrosis, repairable poration, or nonporation) in individual cells. More importantly, our results suggest that membrane poration and cell survival are better correlated with area strain integral (∫​εA2dt) instead of εA,M, which is characteristic of the response of materials under high strain-rate loadings. For 50% cell survival the corresponding area strain integral was found to vary in the range of 56 ∼ 123 μs with εA,M in the range of 57 ∼ 87%. Finally, significant variations in individual cell’s response were observed at the same Sd, indicating the potential for using this method to probe mechanotransduction at the single cell level. PMID:26663913

MEMS squeezer for the measurement of single cell rupture force, stiffness change, and hysteresis

NASA Astrophysics Data System (ADS)

Barazani, B.; Warnat, S.; Fine, A.; Hubbard, T.

2017-02-01

A MEMS squeezer able to compress single living cells underwater until rupture was designed and tested. The relatively large motion range of the device in aqueous media (~2.5 µm) allows provoking cell disruption while measuring cell mechanical properties before and after membrane rupture. An AC driven electrothermal micro actuator with mechanical amplification pressed single cells against a reference back spring. Deformations of the cell and the reference spring were measured with nanoscale resolution using optical Fourier transform techniques. The motion of the reference spring divided by the cell deformation provides the cell stiffness relative to the reference spring constant. An abrupt change in the cell stiffness and the appearance of cracks indicated the cell wall rupture force was reached. A total of 22 baker’s yeast cells (Saccharomyces cerevisiae) were squeezed with the micro device. The average force necessary to rupture the cell membrane was 0.47  ±  0.1 µN. Before rupture the cells had an average stiffness of 9.3  ±  3.1 N m-1 the post-rupture stiffness dropped to 0.94  ±  0.57 N m-1. Cell hysteresis was also measured: cells squeezed and released before reaching the rupture force showed residual deformations below 100 nm, while cells squeezed past the rupture force and then released showed residual deformations between 490 and 990 nm.

Changes in neutrophil morphology and morphometry following exposure to cigarette smoke.

PubMed Central

Lannan, S.; McLean, A.; Drost, E.; Gillooly, M.; Donaldson, K.; Lamb, D.; MacNee, W.

1992-01-01

Acute cigarette smoking delays neutrophils within the pulmonary circulation in some smokers. Evidence from an in-vitro Micropore filter model of the pulmonary capillaries indicates that this may be due to a smoke induced decrease in cell deformability. In order to determine whether changes in cell shape are associated with the observed decrease in neutrophil deformability following smoke exposure, cell morphology, using scanning electron microscopy, and morphometric measurements, made using transmission electron microscopy, were performed on aliquots of neutrophils harvested from whole blood in non-smoking subjects before and after exposure in vitro to cigarette smoke. Smoke exposure increased the maximum diameter and circumference of neutrophils, without changing their area. There was also a change in the maximum to minimum cell diameter ratio, which indicated that the cells had become less spherical. Scanning electron microscopy showed that smoke exposed cells had developed blebbing of their surface membranes, suggestive of an oxidative injury to the cell membrane rather than the shape changes associated with cell activation. These changes in the morphology and morphometry of smoke exposed neutrophils may contribute to the reduction in cell deformability induced by cigarette smoke. Images Fig. 3 Fig. 4 Fig. 5 PMID:1571278

Three-dimensional motion and deformation of a red blood cell in bifurcated microvessels

NASA Astrophysics Data System (ADS)

Ye, Ting; Peng, Lina; Li, Yu

2018-02-01

Microvessels are generally not simple straight tubes, but rather they continually bifurcate (namely, diverging bifurcation) and merge with other microvessels (namely, converging bifurcation). This paper presents a simulation study on the three-dimensional motion and deformation of a red blood cell (RBC) in a bifurcated microvessel with both diverging and converging bifurcations. The motion of the fluids inside and outside of the RBC is modeled by smooth dissipative particle dynamics. The RBC membrane is modeled as a triangular network, having the ability to not only resist the stretching and bending deformations, but also to conserve the RBC volume and surface area. The bifurcation configurations have been studied, including the bifurcated angle and the branch diameter, as well as the RBC properties, including the initial shape, shear modulus, and bending modulus. The simulation results show that the RBC deformation can be divided into five stages, when the RBC flows through a diverging-converging bifurcated microvessel. In these five stages, the RBCs have similar deformation trends but different deformation indices, subject to different bifurcation configurations or different RBC properties. If the shear modulus is large enough, the RBC membrane presents several folds; if the bending modulus is large enough, the RBC loses the symmetry completely with the long shape. These results are helpful in understanding the motion and deformation of healthy or unhealthy cells in blood microcirculation.

High-performance Fuel Cell with Stretched Catalyst-Coated Membrane: One-step Formation of Cracked Electrode.

PubMed

Kim, Sang Moon; Ahn, Chi-Yeong; Cho, Yong-Hun; Kim, Sungjun; Hwang, Wonchan; Jang, Segeun; Shin, Sungsoo; Lee, Gunhee; Sung, Yung-Eun; Choi, Mansoo

2016-05-23

We have achieved performance enhancement of polymer electrolyte membrane fuel cell (PEMFC) though crack generation on its electrodes. It is the first attempt to enhance the performance of PEMFC by using cracks which are generally considered as defects. The pre-defined, cracked electrode was generated by stretching a catalyst-coated Nafion membrane. With the strain-stress property of the membrane that is unique in the aspect of plastic deformation, membrane electrolyte assembly (MEA) was successfully incorporated into the fuel cell. Cracked electrodes with the variation of strain were investigated and electrochemically evaluated. Remarkably, mechanical stretching of catalyst-coated Nafion membrane led to a decrease in membrane resistance and an improvement in mass transport, which resulted in enhanced device performance.

Modeling of Fluid-Membrane Interaction in Cellular Microinjection Process

NASA Astrophysics Data System (ADS)

Karzar-Jeddi, Mehdi; Diaz, Jhon; Olgac, Nejat; Fan, Tai-Hsi

2009-11-01

Cellular microinjection is a well-accepted method to deliver matters such as sperm, nucleus, or macromolecules into biological cells. To improve the success rate of in vitro fertilization and to establish the ideal operating conditions for a novel computer controlled rotationally oscillating intracytoplasmic sperm injection (ICSI) technology, we investigate the fluid-membrane interactions in the ICSI procedure. The procedure consists of anchoring the oocyte (a developing egg) using a holding pipette, penetrating oocyte's zona pellucida (the outer membrane) and the oolemma (the plasma or inner membrane) using an injection micropipette, and finally to deliver sperm into the oocyte for fertilization. To predict the large deformation of the oocyte membranes up to the piercing of the oolemma and the motion of fluids across both membranes, the dynamic fluid-pipette-membrane interactions are formulated by the coupled Stokes' equations and the continuum membrane model based on Helfrich's energy theory. A boundary integral model is developed to simulate the transient membrane deformation and the local membrane stress induced by the longitudinal motion of the injection pipette. The model captures the essential features of the membranes shown on optical images of ICSI experiments, and is capable of suggesting the optimal deformation level of the oolemma to start the rotational oscillations for piercing into the oolemma.

C 60 fullerene localization and membrane interactions in RAW 264.7 immortalized mouse macrophages

DOE Office of Scientific and Technical Information (OSTI.GOV)

Russ, K. A.; Elvati, P.; Parsonage, T. L.

There continues to be a significant increase in the number and complexity of hydrophobic nanomaterials that are engineered for a variety of commercial purposes making human exposure a significant health concern. This study uses a combination of biophysical, biochemical and computational methods to probe potential mechanisms for uptake of C 60 nanoparticles into various compartments of living immune cells. Cultures of RAW 264.7 immortalized murine macrophage were used as a canonical model of immune-competent cells that are likely to provide the first line of defense following inhalation. Modes of entry studied were endocytosis/pinocytosis and passive permeation of cellular membranes. Themore » evidence suggests marginal uptake of C 60 clusters is achieved through endocytosis/pinocytosis, and that passive diffusion into membranes provides a significant source of biologically-available nanomaterial. Compu-tational modeling of both a single molecule and a small cluster of fullerenes predicts that low concentrations of fullerenes enter the membrane individually and produce limited perturbation; however, at higher concentrations the clusters in the membrane causes deformation of the membrane. These findings are bolstered by nuclear magnetic resonance (NMR) of model membranes that reveal defor-mation of the cell membrane upon exposure to high concentrations of fullerenes. The atomistic and NMR models fail to explain escape of the particle out of biological membranes, but are limited to idealized systems that do not completely recapitulate the complexity of cell membranes. Lastly, the surprising contribution of passive modes of cellular entry provides new avenues for toxicological research that go beyond the pharmacological inhibition of bulk transport systems such as pinocytosis.« less

C 60 fullerene localization and membrane interactions in RAW 264.7 immortalized mouse macrophages

DOE PAGES

Russ, K. A.; Elvati, P.; Parsonage, T. L.; ...

2016-01-01

There continues to be a significant increase in the number and complexity of hydrophobic nanomaterials that are engineered for a variety of commercial purposes making human exposure a significant health concern. This study uses a combination of biophysical, biochemical and computational methods to probe potential mechanisms for uptake of C 60 nanoparticles into various compartments of living immune cells. Cultures of RAW 264.7 immortalized murine macrophage were used as a canonical model of immune-competent cells that are likely to provide the first line of defense following inhalation. Modes of entry studied were endocytosis/pinocytosis and passive permeation of cellular membranes. Themore » evidence suggests marginal uptake of C 60 clusters is achieved through endocytosis/pinocytosis, and that passive diffusion into membranes provides a significant source of biologically-available nanomaterial. Compu-tational modeling of both a single molecule and a small cluster of fullerenes predicts that low concentrations of fullerenes enter the membrane individually and produce limited perturbation; however, at higher concentrations the clusters in the membrane causes deformation of the membrane. These findings are bolstered by nuclear magnetic resonance (NMR) of model membranes that reveal defor-mation of the cell membrane upon exposure to high concentrations of fullerenes. The atomistic and NMR models fail to explain escape of the particle out of biological membranes, but are limited to idealized systems that do not completely recapitulate the complexity of cell membranes. Lastly, the surprising contribution of passive modes of cellular entry provides new avenues for toxicological research that go beyond the pharmacological inhibition of bulk transport systems such as pinocytosis.« less

Numerical analysis of a red blood cell flowing through a thin micropore.

PubMed

Omori, Toshihiro; Hosaka, Haruki; Imai, Yohsuke; Yamaguchi, Takami; Ishikawa, Takuji

2014-01-01

Red blood cell (RBC) deformability plays a key role in microcirculation, especially in vessels that have diameters even smaller than the nominal cell size. In this study, we numerically investigate the dynamics of an RBC in a thin micropore. The RBC is modeled as a capsule with a thin hyperelastic membrane. In a numerical simulation, we employ a boundary element method for fluid mechanics and a finite element method for membrane mechanics. The resulting RBC deformation towards the flow direction is suppressed considerably by increased cytoplasm viscosity, whereas the gap between the cell membrane and solid wall becomes smaller with higher cytoplasm viscosity. We also measure the transit time of the RBC and find that nondimensional transit time increases nonlinearly with respect to the viscosity ratio, whereas it is invariant to the capillary number. In conclusion, cytoplasmic viscosity plays a key role in the dynamics of an RBC in a thin pore. The results of this study will be useful for designing a microfluidic device to measure cytoplasmic viscosity.

In situ observation of single cell response to acoustic droplet vaporization: Membrane deformation, permeabilization, and blebbing.

PubMed

Qin, Dui; Zhang, Lei; Chang, Nan; Ni, Pengying; Zong, Yujin; Bouakaz, Ayache; Wan, Mingxi; Feng, Yi

2018-02-06

In this study, the bioeffects of acoustic droplet vaporization (ADV) on adjacent cells were investigated by evaluating the real-time cell response at the single-cell level in situ, using a combined ultrasound-exposure and optical imaging system. Two imaging modalities, high-speed and fluorescence imaging, were used to observe ADV bubble dynamics and to evaluate the impact on cell membrane permeabilization (i.e., sonoporation) using propidium iodide (PI) uptake as an indicator. The results indicated that ADV mainly led to irreversible rather than reversible sonoporation. Further, the rate of irreversible sonoporation significantly increased with increasing nanodroplet concentration, ultrasound amplitude, and pulse duration. The results suggested that sonoporation is correlated to the rapid formation, expansion, and contraction of ADV bubbles near cells, and strongly depends on ADV bubble size and bubble-to-cell distance when subjected to short ultrasound pulses (1 μs). Moreover, the displacement of ADV bubbles was larger when using a long ultrasound pulse (20 μs), resulting in considerable cell membrane deformation and a more irreversible sonoporation rate. During sonoporation, cell membrane blebbing as a recovery manoeuvre was also investigated, indicating the essential role of Ca 2+ influx in the membrane blebbing response. This study has helped us gain further insights into the dynamic behavior of ADV bubbles near cells, ADV bubble-cell interactions, and real-time cell response, which are invaluable in the development of optimal approaches for ADV-associated theranostic applications. Copyright © 2018 Elsevier B.V. All rights reserved.

Red Blood Cell Deformation Under Shear Flow: The Effect of Changing Cell Properties

NASA Astrophysics Data System (ADS)

Forsyth, Alison M.; Wan, Jiandi; Ristenpart, William D.; Stone, Howard A.

2008-11-01

The deformability of red blood cells plays a major role in the pathology of several diseases, including malaria, sickle cell anemia and spherocytosis. Moreover, deformations are believed to trigger the release of adenosine triphosphate, which helps regulate vascular tone and is consequently an important factor in various vascular diseases. Here we investigate single-cell viscoelastic responses to increased shear stress in poly(dimethylsiloxane) channels with a single constriction 2-4 times larger than a typical erythrocyte. These channels mimic arteriole-sized vessels, and have the advantage that the cell membrane is not in contact with the channel walls which have vastly different mechanical and material properties than living tissue. High-speed video and image analysis were used to quantify the trajectories and deformations of cells exposed to varied doses of diamide, a chemical known to ``rigidify'' erythrocytes. Our results show that (i) deformation is proportional to shear rate and (ii) the deformability of diamide-treated cells is greater than that of untreated cells. The latter is an unforeseen result because micropipette aspiration experiments have shown the opposite. We expect that the experimental procedure described here will be useful for characterizing the effect of different therapeutic agents on cellular deformability.

Quantifying the deformation of the red blood cell skeleton in shear flow

NASA Astrophysics Data System (ADS)

Peng, Zhangli; Zhu, Qiang

2012-02-01

To quantitatively predict the response of red blood cell (RBC) membrane in shear flow, we carried out multiphysics simulations by coupling a three-level multiscale approach of RBC membranes with a Boundary Element Method (BEM) for surrounding flows. Our multiscale approach includes a model of spectrins with the domain unfolding feature, a molecular-based model of the junctional complex with detailed protein connectivity and a whole cell Finite Element Method (FEM) model with the bilayer-skeleton friction derived from measured transmembrane protein diffusivity based on the Einstein-Stokes relation. Applying this approach, we investigated the bilayer-skeleton slip and skeleton deformation of healthy RBCs and RBCs with hereditary spherocytosis anemia during tank-treading motion. Compared with healthy cells, cells with hereditary spherocytosis anemia sustain much larger skeleton-bilayer slip and area deformation of the skeleton due to deficiency of transmembrane proteins. This leads to extremely low skeleton density and large bilayer-skeleton interaction force, both of which may cause bilayer loss. This finding suggests a possible mechanism of the development of hereditary spherocytosis anemia.

Coordinated Mechanosensitivity of Membrane Rafts and Focal Adhesions

PubMed Central

Fuentes, Daniela E.; Butler, Peter J.

2013-01-01

Endothelial cells sense mechanical forces of blood flow through mechanisms that involve focal adhesions (FAs). The mechanosensitive pathways that originate from FA-associated integrin activation may involve membrane rafts, small cholesterol- and sphigolipid-rich domains that are either immobilized, by virtue of their attachment to the cytoskeleton, or highly mobile in the plane of the plasma membrane. In this study, we fluorescently labeled non-mobile and mobile populations of GM1, a ganglioside associated with lipid rafts, and transfected cells with the red fluorescent protein-(RFP-) talin, an indicator of integrin activation at FAs, in order to determine the kinetics and sequential order of raft and talin mechanosensitivity. Cells were imaged under confocal microscopy during mechanical manipulation of a FA induced by a fibronectin (FN)-functionalized nanoelectrode with feedback control of position. First, FA deformation led to long range deformation of immobile rafts followed by active recoil of a subpopulation of displaced rafts. Second, initial adhesion between the FN-probe and the cell induced rapid accumulation of GM1 at the probe site with a time constant of 1.7 s. Talin accumulated approximately 20 s later with a time constant of 0.6 s. Third, a 1 μm deformation of the FA lead to immediate (0.3 s) increase in GM1 fluorescence and a later (6 s) increase in talin. Fourth, long term deformation of FAs led to continual GM1 accumulation at the probe site that was reversed upon removal of the deformation. These results demonstrate that rafts are directly mechanosensitive and that raft mobility may enable the earliest events related to FA mechanosensing and reinforcement upon force application. PMID:23487555

Mechanical stability of the cell nucleus: roles played by the cytoskeleton in nuclear deformation and strain recovery.

PubMed

Wang, Xian; Liu, Haijiao; Zhu, Min; Cao, Changhong; Xu, Zhensong; Tsatskis, Yonit; Lau, Kimberly; Kuok, Chikin; Filleter, Tobin; McNeill, Helen; Simmons, Craig A; Hopyan, Sevan; Sun, Yu

2018-05-18

Extracellular forces transmitted through the cytoskeleton can deform the cell nucleus. Large nuclear deformation increases the risk of disrupting the nuclear envelope's integrity and causing DNA damage. Mechanical stability of the nucleus defines its capability of maintaining nuclear shape by minimizing nuclear deformation and recovering strain when deformed. Understanding the deformation and recovery behavior of the nucleus requires characterization of nuclear viscoelastic properties. Here, we quantified the decoupled viscoelastic parameters of the cell membrane, cytoskeleton, and the nucleus. The results indicate that the cytoskeleton enhances nuclear mechanical stability by lowering the effective deformability of the nucleus while maintaining nuclear sensitivity to mechanical stimuli. Additionally, the cytoskeleton decreases the strain energy release rate of the nucleus and might thus prevent shape change-induced structural damage to chromatin. © 2018. Published by The Company of Biologists Ltd.

Phosphatidylinositol 4,5-Bisphosphate (PtdIns(4,5)P2) Specifically Induces Membrane Penetration and Deformation by Bin/Amphiphysin/Rvs (BAR) Domains*

PubMed Central

Yoon, Youngdae; Zhang, Xiuqi; Cho, Wonhwa

2012-01-01

Cellular proteins containing Bin/amphiphysin/Rvs (BAR) domains play a key role in clathrin-mediated endocytosis. Despite extensive structural and functional studies of BAR domains, it is still unknown how exactly these domains interact with the plasma membrane containing phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2) and whether they function by a universal mechanism or by different mechanisms. Here we report that PtdIns(4,5)P2 specifically induces partial membrane penetration of the N-terminal amphiphilic α-helix (H0) of two representative N-BAR domains from Drosophila amphiphysin (dAmp-BAR) and rat endophilin A1 (EndoA1-BAR). Our quantitative fluorescence imaging analysis shows that PtdIns(4,5)P2-dependent membrane penetration of H0 is important for self-association of membrane-bound dAmp-BAR and EndoA1-BAR and their membrane deformation activity. EndoA1-BAR behaves differently from dAmp-BAR because the former has an additional amphiphilic α-helix that penetrates the membrane in a PtdIns(4,5)P2-independent manner. Depletion of PtdIns(4,5)P2 from the plasma membrane of HEK293 cells abrogated the membrane deforming activity of EndoA1-BAR and dAmp-BAR. Collectively, these studies suggest that the local PtdIns(4,5)P2 concentration in the plasma membrane may regulate the membrane interaction and deformation by N-BAR domain-containing proteins during clathrin-mediated endocytosis. PMID:22888025

Influence of the ionophore A23187 on the plastic behavior of normal erythrocytes.

PubMed

Kuettner, J F; Dreher, K L; Rao, G H; Eaton, J W; Blackshear, P L; White, J G

1977-07-01

Previous studies have demonstrated that A23187, an ionophore which selectively transports divalent cations across cell membranes, has profound effects on human erythrocytes: it causes red cells to take up calcium; lose potassium, water, and ATP; convert from biconcave discs to echinocytes and spheroechinocytes; and become more rigid. The present study has explored the influence of calcium uptake induced by the ionophore on the behavior of individual erythrocyte membranes by the micropipette aspiration technique. Exposure of erythrocytes to calcium and A23187 for intervals of up to 30 minutes resulted in marked changes in membrane viscoelastic properties, including the development of increased resistance to aspiration. The most striking manifestation of altered membrane mechanics was apparent after 10 minutes on incubation. Cells pulled into the pipette for a few seconds and the extruded back into the medium retained the deformity imposed by the pipette for several seconds to a few minutes before regaining the form they manifested prior to initial aspiration. The calcium-induced changes in erythrocyte behavior observed in this study strongly support the concept that extrinsic proteins located inside the membrane provide mechanical support to the cell wall, and that increased levels of calcium cause precipitation or cross-linking of the proteins responsible for the increased resistence to deformation and recoil observed after aspiration into micropipettes.

High-performance Fuel Cell with Stretched Catalyst-Coated Membrane: One-step Formation of Cracked Electrode

PubMed Central

Kim, Sang Moon; Ahn, Chi-Yeong; Cho, Yong-Hun; Kim, Sungjun; Hwang, Wonchan; Jang, Segeun; Shin, Sungsoo; Lee, Gunhee; Sung, Yung-Eun; Choi, Mansoo

2016-01-01

We have achieved performance enhancement of polymer electrolyte membrane fuel cell (PEMFC) though crack generation on its electrodes. It is the first attempt to enhance the performance of PEMFC by using cracks which are generally considered as defects. The pre-defined, cracked electrode was generated by stretching a catalyst-coated Nafion membrane. With the strain-stress property of the membrane that is unique in the aspect of plastic deformation, membrane electrolyte assembly (MEA) was successfully incorporated into the fuel cell. Cracked electrodes with the variation of strain were investigated and electrochemically evaluated. Remarkably, mechanical stretching of catalyst-coated Nafion membrane led to a decrease in membrane resistance and an improvement in mass transport, which resulted in enhanced device performance. PMID:27210793

Measurement of in-plane elasticity of live cell layers using a pressure sensor embedded microfluidic device

NASA Astrophysics Data System (ADS)

Lin, Chien-Han; Wang, Chien-Kai; Chen, Yu-An; Peng, Chien-Chung; Liao, Wei-Hao; Tung, Yi-Chung

2016-11-01

In various physiological activities, cells experience stresses along their in-plane direction when facing substrate deformation. Capability of continuous monitoring elasticity of live cell layers during a period is highly desired to investigate cell property variation during various transformations under normal or disease states. This paper reports time-lapsed measurement of live cell layer in-plane elasticity using a pressure sensor embedded microfluidic device. The sensor converts pressure-induced deformation of a flexible membrane to electrical signals. When cells are cultured on top of the membrane, flexural rigidity of the composite membrane increases and further changes the output electrical signals. In the experiments, human embryonic lung fibroblast (MRC-5) cells are cultured and analyzed to estimate the in-plane elasticity. In addition, the cells are treated with a growth factor to simulate lung fibrosis to study the effects of cell transformation on the elasticity variation. For comparison, elasticity measurement on the cells by atomic force microscopy (AFM) is also performed. The experimental results confirm highly anisotropic configuration and material properties of cells. Furthermore, the in-plane elasticity can be monitored during the cell transformation after the growth factor stimulation. Consequently, the developed microfluidic device provides a powerful tool to study physical properties of cells for fundamental biophysics and biomedical researches.

Dietary supplementation with docosahexanoic acid (DHA) increases red blood cell membrane flexibility in mice with sickle cell disease.

PubMed

Wandersee, Nancy J; Maciaszek, Jamie L; Giger, Katie M; Hanson, Madelyn S; Zheng, Suilan; Guo, YiHe; Mickelson, Barbara; Hillery, Cheryl A; Lykotrafitis, George; Low, Philip S; Hogg, Neil

2015-02-01

Humans and mice with sickle cell disease (SCD) have rigid red blood cells (RBCs). Omega-3 fatty acids, such as docosahexanoic acid (DHA), may influence RBC deformability via incorporation into the RBC membrane. In this study, sickle cell (SS) mice were fed natural ingredient rodent diets supplemented with 3% DHA (DHA diet) or a control diet matched in total fat (CTRL diet). After 8weeks of feeding, we examined the RBCs for: 1) stiffness, as measured by atomic force microscopy; 2) deformability, as measured by ektacytometry; and 3) percent irreversibly sickled RBCs on peripheral blood smears. Using atomic force microscopy, it is found that stiffness is increased and deformability decreased in RBCs from SS mice fed CTRL diet compared to wild-type mice. In contrast, RBCs from SS mice fed DHA diet had markedly decreased stiffness and increased deformability compared to RBCs from SS mice fed CTRL diet. Furthermore, examination of peripheral blood smears revealed less irreversibly sickled RBCs in SS mice fed DHA diet as compared to CTRL diet. In summary, our findings indicate that DHA supplementation improves RBC flexibility and reduces irreversibly sickled cells by 40% in SS mice. These results point to potential therapeutic benefits of dietary omega-3 fatty acids in SCD. Copyright © 2014 Elsevier Inc. All rights reserved.

Placing and shaping liposomes with reconfigurable DNA nanocages

NASA Astrophysics Data System (ADS)

Zhang, Zhao; Yang, Yang; Pincet, Frederic; C. Llaguno, Marc; Lin, Chenxiang

2017-07-01

The diverse structure and regulated deformation of lipid bilayer membranes are among a cell's most fascinating features. Artificial membrane-bound vesicles, known as liposomes, are versatile tools for modelling biological membranes and delivering foreign objects to cells. To fully mimic the complexity of cell membranes and optimize the efficiency of delivery vesicles, controlling liposome shape (both statically and dynamically) is of utmost importance. Here we report the assembly, arrangement and remodelling of liposomes with designer geometry: all of which are exquisitely controlled by a set of modular, reconfigurable DNA nanocages. Tubular and toroid shapes, among others, are transcribed from DNA cages to liposomes with high fidelity, giving rise to membrane curvatures present in cells yet previously difficult to construct in vitro. Moreover, the conformational changes of DNA cages drive membrane fusion and bending with predictable outcomes, opening up opportunities for the systematic study of membrane mechanics.

Placing and shaping liposomes with reconfigurable DNA nanocages.

PubMed

Zhang, Zhao; Yang, Yang; Pincet, Frederic; Llaguno, Marc C; Lin, Chenxiang

2017-06-23

The diverse structure and regulated deformation of lipid bilayer membranes are among a cell's most fascinating features. Artificial membrane-bound vesicles, known as liposomes, are versatile tools for modelling biological membranes and delivering foreign objects to cells. To fully mimic the complexity of cell membranes and optimize the efficiency of delivery vesicles, controlling liposome shape (both statically and dynamically) is of utmost importance. Here we report the assembly, arrangement and remodelling of liposomes with designer geometry: all of which are exquisitely controlled by a set of modular, reconfigurable DNA nanocages. Tubular and toroid shapes, among others, are transcribed from DNA cages to liposomes with high fidelity, giving rise to membrane curvatures present in cells yet previously difficult to construct in vitro. Moreover, the conformational changes of DNA cages drive membrane fusion and bending with predictable outcomes, opening up opportunities for the systematic study of membrane mechanics.

Control of cytoskeletal mechanics by extracellular matrix, cell shape, and mechanical tension

NASA Technical Reports Server (NTRS)

Wang, N.; Ingber, D. E.

1994-01-01

We have investigated how extracellular matrix (ECM) alters the mechanical properties of the cytoskeleton (CSK). Mechanical stresses were applied to integrin receptors on the apical surfaces of adherent endothelial cells using RGD-coated ferromagnetic microbeads (5.5-microns diameter) in conjunction with a magnetic twisting device. Increasing the number of basal cell-ECM contacts by raising the fibronectin (FN) coating density from 10 to 500 ng/cm2 promoted cell spreading by fivefold and increased CSK stiffness, apparent viscosity, and permanent deformation all by more than twofold, as measured in response to maximal stress (40 dyne/cm2). When the applied stress was increased from 7 to 40 dyne/cm2, the stiffness and apparent viscosity of the CSK increased in parallel, although cell shape, ECM contacts, nor permanent deformation was altered. Application of the same stresses over a lower number ECM contacts using smaller beads (1.4-microns diameter) resulted in decreased CSK stiffness and apparent viscosity, confirming that this technique probes into the depth of the CSK and not just the cortical membrane. When magnetic measurements were carried out using cells whose membranes were disrupted and ATP stores depleted using saponin, CSK stiffness and apparent viscosity were found to rise by approximately 20%, whereas permanent deformation decreased by more than half. Addition of ATP (250 microM) under conditions that promote CSK tension generation in membrane-permeabilized cells resulted in decreases in CSK stiffness and apparent viscosity that could be detected within 2 min after ATP addition, before any measurable change in cell size.(ABSTRACT TRUNCATED AT 250 WORDS).

A computational model of amoeboid cell swimming in unbounded medium and through obstacles

NASA Astrophysics Data System (ADS)

Campbell, Eric; Bagchi, Prosenjit

2017-11-01

Pseudopod-driven motility is commonly observed in eukaryotic cells. Pseudopodia are actin-rich protrusions of the cellular membrane which extend, bifurcate, and retract in cycles resulting in amoeboid locomotion. While actin-myosin interactions are responsible for pseudopod generation, cell deformability is crucial concerning pseudopod dynamics. Because pseudopodia are highly dynamic, cells are capable of deforming into complex shapes over time. Pseudopod-driven motility represents a multiscale and complex process, coupling cell deformation, protein biochemistry, and cytoplasmic and extracellular fluid motion. In this work, we present a 3D computational model of amoeboid cell swimming in an extracellular medium (ECM). The ECM is represented as a fluid medium with or without obstacles. The model integrates full cell deformation, a coarse-grain reaction-diffusion system for protein dynamics, and fluid interaction. Our model generates pseudopodia which bifurcate and retract, showing remarkable similarity to experimental observations. Influence of cell deformation, protein diffusivity and cytoplasmic viscosity on the swimming speed is analyzed in terms of altered pseudopod dynamics. Insights into the role of matrix porosity and obstacle size on cell motility are also provided. Funded by NSF CBET 1438255.

High-throughput linear optical stretcher for mechanical characterization of blood cells.

PubMed

Roth, Kevin B; Neeves, Keith B; Squier, Jeff; Marr, David W M

2016-04-01

This study describes a linear optical stretcher as a high-throughput mechanical property cytometer. Custom, inexpensive, and scalable optics image a linear diode bar source into a microfluidic channel, where cells are hydrodynamically focused into the optical stretcher. Upon entering the stretching region, antipodal optical forces generated by the refraction of tightly focused laser light at the cell membrane deform each cell in flow. Each cell relaxes as it flows out of the trap and is compared to the stretched state to determine deformation. The deformation response of untreated red blood cells and neutrophils were compared to chemically treated cells. Statistically significant differences were observed between normal, diamide-treated, and glutaraldehyde-treated red blood cells, as well as between normal and cytochalasin D-treated neutrophils. Based on the behavior of the pure, untreated populations of red cells and neutrophils, a mixed population of these cells was tested and the discrete populations were identified by deformability. © 2015 International Society for Advancement of Cytometry. © 2015 International Society for Advancement of Cytometry.

Fractal avalanche ruptures in biological membranes

NASA Astrophysics Data System (ADS)

Gözen, Irep; Dommersnes, Paul; Czolkos, Ilja; Jesorka, Aldo; Lobovkina, Tatsiana; Orwar, Owe

2010-11-01

Bilayer membranes envelope cells as well as organelles, and constitute the most ubiquitous biological material found in all branches of the phylogenetic tree. Cell membrane rupture is an important biological process, and substantial rupture rates are found in skeletal and cardiac muscle cells under a mechanical load. Rupture can also be induced by processes such as cell death, and active cell membrane repair mechanisms are essential to preserve cell integrity. Pore formation in cell membranes is also at the heart of many biomedical applications such as in drug, gene and short interfering RNA delivery. Membrane rupture dynamics has been studied in bilayer vesicles under tensile stress, which consistently produce circular pores. We observed very different rupture mechanics in bilayer membranes spreading on solid supports: in one instance fingering instabilities were seen resulting in floral-like pores and in another, the rupture proceeded in a series of rapid avalanches causing fractal membrane fragmentation. The intermittent character of rupture evolution and the broad distribution in avalanche sizes is consistent with crackling-noise dynamics. Such noisy dynamics appear in fracture of solid disordered materials, in dislocation avalanches in plastic deformations and domain wall magnetization avalanches. We also observed similar fractal rupture mechanics in spreading cell membranes.

On the shape memory of red blood cells

NASA Astrophysics Data System (ADS)

Cordasco, Daniel; Bagchi, Prosenjit

2017-04-01

Red blood cells (RBCs) undergo remarkably large deformations when subjected to external forces but return to their biconcave discoid resting shape as the forces are withdrawn. In many experiments, such as when RBCs are subjected to a shear flow and undergo the tank-treading motion, the membrane elements are also displaced from their original (resting) locations along the cell surface with respect to the cell axis, in addition to the cell being deformed. A shape memory is said to exist if after the flow is stopped the RBC regains its biconcave shape and the membrane elements also return to their original locations. The shape memory of RBCs was demonstrated by Fischer ["Shape memory of human red blood cells," Biophys. J. 86, 3304-3313 (2004)] using shear flow go-and-stop experiments. Optical tweezer and micropipette based stretch-relaxation experiments do not reveal the complete shape memory because while the RBC may be deformed, the membrane elements are not significantly displaced from their original locations with respect to the cell axis. Here we present the first three-dimensional computational study predicting the complete shape memory of RBCs using shear flow go-and-stop simulations. The influence of different parameters, namely, membrane shear elasticity and bending rigidity, membrane viscosity, cytoplasmic and suspending fluid viscosity, as well as different stress-free states of the RBC is studied. For all cases, the RBCs always exhibit shape memory. The complete recovery of the RBC in shear flow go-and-stop simulations occurs over a time that is orders of magnitude longer than that for optical tweezer and micropipette based relaxations. The response is also observed to be more complex and composed of widely disparate time scales as opposed to only one time scale that characterizes the optical tweezer and micropipette based relaxations. We observe that the recovery occurs in three phases: a rapid compression of the RBC immediately after the flow is stopped, followed by a slow recovery to the biconcave shape combined with membrane rotation, and a final rotational return of the membrane elements back to their original locations. A fast time scale on the order of a few hundred milliseconds characterizes the initial compression phase while a slow time scale on the order of tens of seconds is associated with the rotational phase. We observe that the response is strongly dependent on the stress-free state of the cells, that is, the relaxation time decreases significantly and the mode of recovery changes from rotation-driven to deformation-driven as the stress-free state becomes more non-spherical. We show that while membrane shear elasticity and non-spherical stress-free shape are necessary and sufficient for the membrane elements to return to their original locations, bending rigidity is needed for the "global" recovery of the biconcave shape. We also perform a novel relaxation simulation in which the cell axis of revolution is not aligned with the shear plane and show that the shape memory is exhibited even when the membrane elements are displaced normal to the imposed flow direction. The results presented here could motivate new experiments to determine the exact stress-free state of the RBC and also to clearly identify different tank-treading modes.

Red blood cell oxidative stress impairs oxygen delivery and induces red blood cell aging.

PubMed

Mohanty, Joy G; Nagababu, Enika; Rifkind, Joseph M

2014-01-01

Red Blood Cells (RBCs) need to deform and squeeze through narrow capillaries. Decreased deformability of RBCs is, therefore, one of the factors that can contribute to the elimination of aged or damaged RBCs from the circulation. This process can also cause impaired oxygen delivery, which contributes to the pathology of a number of diseases. Studies from our laboratory have shown that oxidative stress plays a significant role in damaging the RBC membrane and impairing its deformability. RBCs are continuously exposed to both endogenous and exogenous sources of reactive oxygen species (ROS) like superoxide and hydrogen peroxide (H2O2). The bulk of the ROS are neutralized by the RBC antioxidant system consisting of both non-enzymatic and enzymatic antioxidants including catalase, glutathione peroxidase and peroxiredoxin-2. However, the autoxidation of hemoglobin (Hb) bound to the membrane is relatively inaccessible to the predominantly cytosolic RBC antioxidant system. This inaccessibility becomes more pronounced under hypoxic conditions when Hb is partially oxygenated, resulting in an increased rate of autoxidation and increased affinity for the RBC membrane. We have shown that a fraction of peroxyredoxin-2 present on the RBC membrane may play a major role in neutralizing these ROS. H2O2 that is not neutralized by the RBC antioxidant system can react with the heme producing fluorescent heme degradation products (HDPs). We have used the level of these HDP as a measure of RBC oxidative Stress. Increased levels of HDP are detected during cellular aging and various diseases. The negative correlation (p < 0.0001) between the level of HDP and RBC deformability establishes a contribution of RBC oxidative stress to impaired deformability and cellular stiffness. While decreased deformability contributes to the removal of RBCs from the circulation, oxidative stress also contributes to the uptake of RBCs by macrophages, which plays a major role in the removal of RBCs from circulation. The contribution of oxidative stress to the removal of RBCs by macrophages involves caspase-3 activation, which requires oxidative stress. RBC oxidative stress, therefore, plays a significant role in inducing RBC aging.

Surface deformation during an action potential in pearled cells

NASA Astrophysics Data System (ADS)

Mussel, Matan; Fillafer, Christian; Ben-Porath, Gal; Schneider, Matthias F.

2017-11-01

Electric pulses in biological cells (action potentials) have been reported to be accompanied by a propagating cell-surface deformation with a nanoscale amplitude. Typically, this cell surface is covered by external layers of polymer material (extracellular matrix, cell wall material, etc.). It was recently demonstrated in excitable plant cells (Chara braunii) that the rigid external layer (cell wall) hinders the underlying deformation. When the cell membrane was separated from the cell wall by osmosis, a mechanical deformation, in the micrometer range, was observed upon excitation of the cell. The underlying mechanism of this mechanical pulse has, to date, remained elusive. Herein we report that Chara cells can undergo a pearling instability, and when the pearled fragments were excited even larger and more regular cell shape changes were observed (˜10 -100 μ m in amplitude). These transient cellular deformations were captured by a curvature model that is based on three parameters: surface tension, bending rigidity, and pressure difference across the surface. In this paper these parameters are extracted by curve-fitting to the experimental cellular shapes at rest and during excitation. This is a necessary step to identify the mechanical parameters that change during an action potential.

Viscoelastic deformation of lipid bilayer vesicles.

PubMed

Wu, Shao-Hua; Sankhagowit, Shalene; Biswas, Roshni; Wu, Shuyang; Povinelli, Michelle L; Malmstadt, Noah

2015-10-07

Lipid bilayers form the boundaries of the cell and its organelles. Many physiological processes, such as cell movement and division, involve bending and folding of the bilayer at high curvatures. Currently, bending of the bilayer is treated as an elastic deformation, such that its stress-strain response is independent of the rate at which bending strain is applied. We present here the first direct measurement of viscoelastic response in a lipid bilayer vesicle. We used a dual-beam optical trap (DBOT) to stretch 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) giant unilamellar vesicles (GUVs). Upon application of a step optical force, the vesicle membrane deforms in two regimes: a fast, instantaneous area increase, followed by a much slower stretching to an eventual plateau deformation. From measurements of dozens of GUVs, the average time constant of the slower stretching response was 0.225 ± 0.033 s (standard deviation, SD). Increasing the fluid viscosity did not affect the observed time constant. We performed a set of experiments to rule out heating by laser absorption as a cause of the transient behavior. Thus, we demonstrate here that the bending deformation of lipid bilayer membranes should be treated as viscoelastic.

Viscoelastic deformation of lipid bilayer vesicles†

PubMed Central

Wu, Shao-Hua; Sankhagowit, Shalene; Biswas, Roshni; Wu, Shuyang; Povinelli, Michelle L.

2015-01-01

Lipid bilayers form the boundaries of the cell and its organelles. Many physiological processes, such as cell movement and division, involve bending and folding of the bilayer at high curvatures. Currently, bending of the bilayer is treated as an elastic deformation, such that its stress-strain response is independent of the rate at which bending strain is applied. We present here the first direct measurement of viscoelastic response in a lipid bilayer vesicle. We used a dual-beam optical trap (DBOT) to stretch 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) giant unilamellar vesicles (GUVs). Upon application of a step optical force, the vesicle membrane deforms in two regimes: a fast, instantaneous area increase, followed by a much slower stretching to an eventual plateau deformation. From measurements of dozens of GUVs, the average time constant of the slower stretching response was 0.225 ± 0.033 s (standard deviation, SD). Increasing the fluid viscosity did not affect the observed time constant. We performed a set of experiments to rule out heating by laser absorption as a cause of the transient behavior. Thus, we demonstrate here that the bending deformation of lipid bilayer membranes should be treated as viscoelastic. PMID:26268612

Superior Robust Ultrathin Single-Crystalline Silicon Carbide Membrane as a Versatile Platform for Biological Applications.

PubMed

Nguyen, Tuan-Khoa; Phan, Hoang-Phuong; Kamble, Harshad; Vadivelu, Raja; Dinh, Toan; Iacopi, Alan; Walker, Glenn; Hold, Leonie; Nguyen, Nam-Trung; Dao, Dzung Viet

2017-12-06

Micromachined membranes are promising platforms for cell culture thanks to their miniaturization and integration capabilities. Possessing chemical inertness, biocompatibility, and integration, silicon carbide (SiC) membranes have attracted great interest toward biological applications. In this paper, we present the batch fabrication, mechanical characterizations, and cell culture demonstration of robust ultrathin epitaxial deposited SiC membranes. The as-fabricated ultrathin SiC membranes, with an ultrahigh aspect ratio (length/thickness) of up to 20 000, possess high a fracture strength up to 2.95 GPa and deformation up to 50 μm. A high optical transmittance of above 80% at visible wavelengths was obtained for 50 nm membranes. The as-fabricated membranes were experimentally demonstrated as an excellent substrate platform for bio-MEMS/NEMS cell culture with the cell viability rate of more than 92% after 72 h. The ultrathin SiC membrane is promising for in vitro observations/imaging of bio-objects with an extremely short optical access.

A viable circulating tumor cell isolation device with high retrieval efficiency using a reversibly deformable membrane barrier

NASA Astrophysics Data System (ADS)

Kim, Yoonji; Bu, Jiyoon; Cho, Young-Ho; Son, Il Tae; Kang, Sung-Bum

2017-02-01

Circulating tumor cells (CTCs) contain prognostic information of the tumor, since they shed from the primary tumor and invade into the bloodstream. Therefore, the viable isolation is necessary for a consequent analysis of CTCs. Here, we present a device for the viable isolation and efficient retrieval of CTCs using slanted slot filters, formed by a reversibly deformable membrane barrier. Conventional filters have difficulties in retrieving captured cells, since they easily clog the slots. Moreover, large stress concentration at the sharp edges of squared slots, causes cell lysis. In contrast, the present device shows over 94% of high retrieval efficiency, since the slots can be opened simply by relieving the pressure. Furthermore, the inflated membrane barrier naturally forms the slanted slots, thus reducing the cell damage. By using cancer cell lines, we verified that the present device successfully isolate targeted cells, even at an extremely low concentrations (~10 cells/0.1 ml). In the clinical study, 85.7% of patients initially showed CTC positive while the numbers generally decreased after the surgery. We have also proved that the number of CTCs were highly correlated with tumour invasiveness. Therefore, the present device has potential for use in cancer diagnosis, surgical validation, and invasiveness analysis.

Integrated strain array for cellular mechanobiology studies

NASA Astrophysics Data System (ADS)

Simmons, C. S.; Sim, J. Y.; Baechtold, P.; Gonzalez, A.; Chung, C.; Borghi, N.; Pruitt, B. L.

2011-05-01

We have developed an integrated strain array for cell culture enabling high-throughput mechano-transduction studies. Biocompatible cell culture chambers were integrated with an acrylic pneumatic compartment and microprocessor-based control system. Each element of the array consists of a deformable membrane supported by a cylindrical pillar within a well. For user-prescribed waveforms, the annular region of the deformable membrane is pulled into the well around the pillar under vacuum, causing the pillar-supported region with cultured cells to be stretched biaxially. The optically clear device and pillar-based mechanism of operation enables imaging on standard laboratory microscopes. Straightforward fabrication utilizes off-the-shelf components, soft lithography techniques in polydimethylsiloxane and laser ablation of acrylic sheets. Proof of compatibility with basic biological assays and standard imaging equipment were accomplished by straining C2C12 skeletal myoblasts on the device for 6 h. At higher strains, cells and actin stress fibers realign with a circumferential preference.

A computational model of amoeboid cell swimming

NASA Astrophysics Data System (ADS)

Campbell, Eric J.; Bagchi, Prosenjit

2017-10-01

Amoeboid cells propel by generating pseudopods that are finger-like protrusions of the cell body that continually grow, bifurcate, and retract. Pseudopod-driven motility of amoeboid cells represents a complex and multiscale process that involves bio-molecular reactions, cell deformation, and cytoplasmic and extracellular fluid motion. Here we present a 3D model of pseudopod-driven swimming of an amoeba suspended in a fluid without any adhesion and in the absence of any chemoattractant. Our model is based on front-tracking/immersed-boundary methods, and it combines large deformation of the cell, a coarse-grain model for molecular reactions, and cytoplasmic and extracellular fluid flow. The predicted shapes of the swimming cell from our model show similarity with experimental observations. We predict that the swimming behavior changes from random-like to persistent unidirectional motion, and that the swimming speed increases, with increasing cell deformability and protein diffusivity. The unidirectionality in cell swimming is observed without any external cues and as a direct result of a change in pseudopod dynamics. We find that pseudopods become preferentially focused near the front of the cell and appear in greater numbers with increasing cell deformability and protein diffusivity, thereby increasing the swimming speed and making the cell shape more elongated. We find that the swimming speed is minimum when the cytoplasm viscosity is close to the extracellular fluid viscosity. We further find that the speed increases significantly as the cytoplasm becomes less viscous compared with the extracellular fluid, resembling the viscous fingering phenomenon observed in interfacial flows. While these results support the notion that softer cells migrate more aggressively, they also suggest a strong coupling between membrane elasticity, membrane protein diffusivity, and fluid viscosity.

Capillarity-induced folds fuel extreme shape changes in thin wicked membranes.

PubMed

Grandgeorge, Paul; Krins, Natacha; Hourlier-Fargette, Aurélie; Laberty-Robert, Christel; Neukirch, Sébastien; Antkowiak, Arnaud

2018-04-20

Soft deformable materials are needed for applications such as stretchable electronics, smart textiles, or soft biomedical devices. However, the design of a durable, cost-effective, or biologically compatible version of such a material remains challenging. Living animal cells routinely cope with extreme deformations by unfolding preformed membrane reservoirs available in the form of microvilli or membrane folds. We synthetically mimicked this behavior by creating nanofibrous liquid-infused tissues that spontaneously form similar reservoirs through capillarity-induced folding. By understanding the physics of membrane buckling within the liquid film, we developed proof-of-concept conformable chemical surface treatments and stretchable basic electronic circuits. Copyright © 2018 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.

Improved performance of single-chamber microbial fuel cells through control of membrane deformation.

PubMed

Zhang, Xiaoyuan; Cheng, Shaoan; Huang, Xia; Logan, Bruce E

2010-03-15

Cation (CEMs) and anion exchange membrane (AEMs) are commonly used in microbial fuel cells (MFCs) to enhance Coulombic efficiencies (CEs) by reducing the flux of oxygen through the cathode to bacteria on the anode. AEMs typically work better than CEMs, but in initial experiments we observed the opposite using a membrane electrode assembly MFC. The reason was identified to be membrane deformation, which resulted in water and gas trapped between the membrane and cathode. To correct this, stainless steel mesh was used to press the membrane flat against the cathode. With the steel mesh, AEM performance increased to 46+/-4 W/m(3) in a single cathode MFC, and 98+/-14 W/m(3) in a double-cathode MFC. These power densities were higher than those using a CEM of 32+/-2 W/m(3) (single cathode) and 63+/-6 W/m(3) (double cathode). Higher pH gradients across the membrane and salt precipitation on the cathode were responsible for the reduced performance of the CEM compared to the AEM. CEs reached over 90% for both membranes at >2A/m(2). These results demonstrate the importance of avoiding water accumulation in thin films between membranes and electrodes, and explain additional reasons for poorer performance of CEMs compared to AEMs. (c) 2009 Elsevier B.V. All rights reserved.

The Role of Membrane Curvature in Nanoscale Topography-Induced Intracellular Signaling.

PubMed

Lou, Hsin-Ya; Zhao, Wenting; Zeng, Yongpeng; Cui, Bianxiao

2018-05-15

Over the past decade, there has been growing interest in developing biosensors and devices with nanoscale and vertical topography. Vertical nanostructures induce spontaneous cell engulfment, which enhances the cell-probe coupling efficiency and the sensitivity of biosensors. Although local membranes in contact with the nanostructures are found to be fully fluidic for lipid and membrane protein diffusions, cells appear to actively sense and respond to the surface topography presented by vertical nanostructures. For future development of biodevices, it is important to understand how cells interact with these nanostructures and how their presence modulates cellular function and activities. How cells recognize nanoscale surface topography has been an area of active research for two decades before the recent biosensor works. Extensive studies show that surface topographies in the range of tens to hundreds of nanometers can significantly affect cell functions, behaviors, and ultimately the cell fate. For example, titanium implants having rough surfaces are better for osteoblast attachment and host-implant integration than those with smooth surfaces. At the cellular level, nanoscale surface topography has been shown by a large number of studies to modulate cell attachment, activity, and differentiation. However, a mechanistic understanding of how cells interact and respond to nanoscale topographic features is still lacking. In this Account, we focus on some recent studies that support a new mechanism that local membrane curvature induced by nanoscale topography directly acts as a biochemical signal to induce intracellular signaling, which we refer to as the curvature hypothesis. The curvature hypothesis proposes that some intracellular proteins can recognize membrane curvatures of a certain range at the cell-to-material interface. These proteins then recruit and activate downstream components to modulate cell signaling and behavior. We discuss current technologies allowing the visualization of membrane deformation at the cell membrane-to-substrate interface with nanometer precision and demonstrate that vertical nanostructures induce local curvatures on the plasma membrane. These local curvatures enhance the process of clathrin-mediated endocytosis and affect actin dynamics. We also present evidence that vertical nanostructures can induce significant deformation of the nuclear membrane, which can affect chromatin distribution and gene expression. Finally, we provide a brief perspective on the curvature hypothesis and the challenges and opportunities for the design of nanotopography for manipulating cell behavior.

Coupled elasticity–diffusion model for the effects of cytoskeleton deformation on cellular uptake of cylindrical nanoparticles

PubMed Central

Wang, Jizeng; Li, Long

2015-01-01

Molecular dynamic simulations and experiments have recently demonstrated how cylindrical nanoparticles (CNPs) with large aspect ratios penetrate animal cells and inevitably deform cytoskeletons. Thus, a coupled elasticity–diffusion model was adopted to elucidate this interesting biological phenomenon by considering the effects of elastic deformations of cytoskeleton and membrane, ligand–receptor binding and receptor diffusion. The mechanism by which the binding energy drives the CNPs with different orientations to enter host cells was explored. This mechanism involved overcoming the resistance caused by cytoskeleton and membrane deformations and the change in configurational entropy of the ligand–receptor bonds and free receptors. Results showed that deformation of the cytoskeleton significantly influenced the engulfing process by effectively slowing down and even hindering the entry of the CNPs. Additionally, the engulfing depth was determined quantitatively. CNPs preferred or tended to vertically attack target cells until they were stuck in the cytoskeleton as implied by the speed of vertically oriented CNPs that showed much faster initial engulfing speeds than horizontally oriented CNPs. These results elucidated the most recent molecular dynamics simulations and experimental observations on the cellular uptake of carbon nanotubes and phagocytosis of filamentous Escherichia coli bacteria. The most efficient engulfment showed the stiffness-dependent optimal radius of the CNPs. Cytoskeleton stiffness exhibited more significant influence on the optimal sizes of the vertical uptake than the horizontal uptake. PMID:25411410

Intravacuolar Membranes Regulate CD8 T Cell Recognition of Membrane-Bound Toxoplasma gondii Protective Antigen.

PubMed

Lopez, Jodie; Bittame, Amina; Massera, Céline; Vasseur, Virginie; Effantin, Grégory; Valat, Anne; Buaillon, Célia; Allart, Sophie; Fox, Barbara A; Rommereim, Leah M; Bzik, David J; Schoehn, Guy; Weissenhorn, Winfried; Dubremetz, Jean-François; Gagnon, Jean; Mercier, Corinne; Cesbron-Delauw, Marie-France; Blanchard, Nicolas

2015-12-15

Apicomplexa parasites such as Toxoplasma gondii target effectors to and across the boundary of their parasitophorous vacuole (PV), resulting in host cell subversion and potential presentation by MHC class I molecules for CD8 T cell recognition. The host-parasite interface comprises the PV limiting membrane and a highly curved, membranous intravacuolar network (IVN) of uncertain function. Here, using a cell-free minimal system, we dissect how membrane tubules are shaped by the parasite effectors GRA2 and GRA6. We show that membrane association regulates access of the GRA6 protective antigen to the MHC I pathway in infected cells. Although insertion of GRA6 in the PV membrane is key for immunogenicity, association of GRA6 with the IVN limits presentation and curtails GRA6-specific CD8 responses in mice. Thus, membrane deformations of the PV regulate access of antigens to the MHC class I pathway, and the IVN may play a role in immune modulation. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

Micro-scale blood particulate dynamics using a non-uniform rational B-spline-based isogeometric analysis.

PubMed

Chivukula, V; Mousel, J; Lu, J; Vigmostad, S

2014-12-01

The current research presents a novel method in which blood particulates - biconcave red blood cells (RBCs) and spherical cells are modeled using isogeometric analysis, specifically Non-Uniform Rational B-Splines (NURBS) in 3-D. The use of NURBS ensures that even with a coarse representation, the geometry of the blood particulates maintains an accurate description when subjected to large deformations. The fundamental advantage of this method is the coupling of the geometrical description and the stress analysis of the cell membrane into a single, unified framework. Details on the modeling approach, implementation of boundary conditions and the membrane mechanics analysis using isogeometric modeling are presented, along with validation cases for spherical and biconcave cells. Using NURBS - based isogeometric analysis, the behavior of individual cells in fluid flow is presented and analyzed in different flow regimes using as few as 176 elements for a spherical cell and 220 elements for a biconcave RBC. This work provides a framework for modeling a large number of 3-D deformable biological cells, each with its own geometric description and membrane properties. To the best knowledge of the authors, this is the first application of the NURBS - based isogeometric analysis to model and simulate blood particulates in flow in 3D. Copyright © 2014 John Wiley & Sons, Ltd.

On the mechanics of elastic lines in thin shells

NASA Astrophysics Data System (ADS)

Benet, Eduard; Vernerey, Franck

The deformation of soft shells in nature and engineering is often conditioned by the presence of lines whose mechanical properties are different from the shell. For instance, the deformation of tree leaves is conditioned by the presence of harder stems, and cell mitosis is driven by a stiffening line along its membrane. From an experimental standpoint, many groups have taken advantage of this feature to develop self-actuated shells with prescribed deformations. Examples include the polymerization of gels along certain lines, or the inclusion of stiffer lines via 3D printing. However, there is not yet a general continuum theory that accounts for this type of discontinuity within the membrane. Hence, we extend the general shell theory to account for the inclusion of a line that potentially induces jumps in stresses, couple stresses and moments, across its thickness. This is achieved via coupling the rod and the membrane deformations, and ensuring continuity of displacements. The model is then applied to three important problems: a constriction disc inside a shell of revolution, the induced twisting of a shell via the torsion of an embedded line, and the effect of an helicoidal line on the uni-axial deformation of a cylindrical shell. National Science Foundation CAREER award 1350090.

Computational model for amoeboid motion: Coupling membrane and cytosol dynamics

NASA Astrophysics Data System (ADS)

Moure, Adrian; Gomez, Hector

2016-10-01

A distinguishing feature of amoeboid motion is that the migrating cell undergoes large deformations, caused by the emergence and retraction of actin-rich protrusions, called pseudopods. Here, we propose a cell motility model that represents pseudopod dynamics, as well as its interaction with membrane signaling molecules. The model accounts for internal and external forces, such as protrusion, contraction, adhesion, surface tension, or those arising from cell-obstacle contacts. By coupling the membrane and cytosol interactions we are able to reproduce a realistic picture of amoeboid motion. The model results are in quantitative agreement with experiments and show how cells may take advantage of the geometry of their microenvironment to migrate more efficiently.

Puncture mechanics of soft elastomeric membrane with large deformation by rigid cylindrical indenter

NASA Astrophysics Data System (ADS)

Liu, Junjie; Chen, Zhe; Liang, Xueya; Huang, Xiaoqiang; Mao, Guoyong; Hong, Wei; Yu, Honghui; Qu, Shaoxing

2018-03-01

Soft elastomeric membrane structures are widely used and commonly found in engineering and biological applications. Puncture is one of the primary failure modes of soft elastomeric membrane at large deformation when indented by rigid objects. In order to investigate the puncture failure mechanism of soft elastomeric membrane with large deformation, we study the deformation and puncture failure of silicone rubber membrane that results from the continuous axisymmetric indentation by cylindrical steel indenters experimentally and analytically. In the experiment, effects of indenter size and the friction between the indenter and the membrane on the deformation and puncture failure of the membrane are investigated. In the analytical study, a model within the framework of nonlinear field theory is developed to describe the large local deformation around the punctured area, as well as to predict the puncture failure of the membrane. The deformed membrane is divided into three parts and the friction contact between the membrane and indenter is modeled by Coulomb friction law. The first invariant of the right Cauchy-Green deformation tensor I1 is adopted to predict the puncture failure of the membrane. The experimental and analytical results agree well. This work provides a guideline in designing reliable soft devices featured with membrane structures, which are present in a wide variety of applications.

Interaction between bending and tension forces in bilayer membranes.

PubMed Central

Secomb, T W

1988-01-01

A theoretical analysis is presented of the bending mechanics of a membrane consisting of two tightly-coupled leaflets, each of which shears and bends readily but strongly resists area changes. Structures of this type have been proposed to model biological membranes such as red blood cell membrane. It is shown that when such a membrane is bent, anisotropic components of resultant membrane tension (shear stresses) are induced, even when the tension in each leaflet is isotropic. The induced shear stresses increase as the square of the membrane curvature, and become significant for moderate curvatures (when the radius of curvature is much larger than the distance between the leaflets). This effect has implications for the analysis of shape and deformation of freely suspended and flowing red blood cells. PMID:3224154

The effect of abnormal hemoglobins on the membrane regulation of cell hydration.

PubMed

Clark, M R; Shohet, S B

Several hemoglobinopathies are associated with abnormalities in the permeability of the red cell membrane, in some cases leading to permanent alterations of the intracellular milieu. Homozygous sickle cell disease is the most thoroughly studied example. Deoxygenation of sickle cells causes a transient increase in the permeability to monovalent cations and Ca; prolonged deoxygenation can lead to a permanent accumulation of Ca and loss of total cations and water. Although the mechanisms for the permeability changes are not yet defined, mechanical stress on the membrane, with subsequent damages by excess Ca or membrane-associated hemoglobin have been suggested to play a role. Loss of cell water and increase in mean cell hemoglobin concentration causes massive reduction of cell deformability in the oxygenated state and makes the hemoglobin more likely to undergo sickling because of the strong concentration dependence of the sickling process. Limited evidence suggests the occurrence of permeability defects in other hemoglobinopathies and the thalassemias. The suggested alterations range from a slight increase in K permeability of incubated thalassemia cells to substantial dehydration of cells from patients with homozygous hemoglobin C disease. Oxidative damage to the membrane, involving an abnormal hemoglobin-membrane association, may underly the permeability changes in these cells.

Membranes, mechanics, and intracellular transport

NASA Astrophysics Data System (ADS)

Parthasarathy, Raghuveer

2012-10-01

Cellular membranes are remarkable materials -- self-assembled, flexible, two-dimensional fluids. Understanding how proteins manipulate membrane curvature is crucial to understanding the transport of cargo in cells, yet the mechanical activities of trafficking proteins remain poorly understood. Using an optical-trap based assay involving dynamic deformation of biomimetic membranes, we have examined the behavior of Sar1, a key component of the COPII family of transport proteins. We find that Sar1 from yeast (S. cerevisiae) lowers membrane rigidity by up to 100% as a function of its concentration, thereby lowering the energetic cost of membrane deformation. Human Sar1 proteins can also lower the mechanical rigidity of the membranes to which they bind. However, unlike the yeast proteins, the rigidity is not a monotonically decreasing function of concentration but rather shows increased rigidity and decreased mobility at high concentrations that implies interactions between proteins. In addition to describing this study of membrane mechanics, I'll also discuss some topics relevant to a range of biophysical investigations, such as the insights provided by imaging methods and open questions in the dynamics of multicellular systems.

[Modification of red cell membranes with perftoran in papaine emphysema in rats].

PubMed

Zoirova, N I; Arifkhanov, S I; Rakhmatullaev, Kh U; Tadzhikhodzhaev, Iu Kh

2006-01-01

Papaine emphysema model on 75 mongrel mature white male rats (10 intact rats were control) was used to study the size, form, surface architechtonics, deformability and state of membrane-receptor erythrocyte complex before and after perftoran intraperitoneal administration. Perftoran emulsion produced a membrane-modulating effect with recovery of hormonal reception sensitivity, PHA-, cAMP-receptor systems as well as restoration of erythrocytic normocytosis and diskocytosis.

Microhydrodynamics of deformable particles: surprising responses of drops and vesicles to uniform electric field or shear flow

NASA Astrophysics Data System (ADS)

Vlahovska, Petia

2015-11-01

Particle motion in a viscous fluid is a classic problem that continues to surprise researchers. In this talk, I will discuss some intriguing, experimentally-observed behaviors of droplets and giant vesicles (cell-size lipid membrane sacs) in electric or flow fields. In a uniform electric field, a droplet deforms into an ellipsoid that can either be steadily tilted relative to the applied field direction or undergo unsteady motions (periodic shape oscillations or irregular flipping); a spherical vesicle can adopt a transient square shape or reversibly porate. In a steady shear flow, a vesicle can tank-tread, tumble or swing. Theoretical models show that the nonlinear drop dynamics originates from the interplay of Quincke rotation and interface deformation, while the vesicle dynamics stems from the membrane inextensibility. The practical motivation for this research lies in an improved understanding of technologies that rely on the manipulation of drops and cells by flow or electric fields.

The effects of non-Newtonian viscosity on the deformation of red blood cells in a shear flow

NASA Astrophysics Data System (ADS)

Sesay, Juldeh

2005-11-01

The analyses of the effects of non-Newtonian viscosity on the membrane of red blood cells (RBCs) suspended in a shear flow are presented. The specific objective is to investigate the mechanical deformation on the surfaces of an ellipsoidal particle model. The hydrodynamic stresses and other forces on the surface of the particle are used to determine the cell deformation. We extended previous works, which were based on the Newtonian fluid models, to the non-Newtonian case, and focus on imposed shear rate values between 1 and 100 per second. Two viscosity models are investigated, which respectively correspond to a normal person and a patient with cerebrovascular accident (CVA). The results are compared with those obtained assuming a Newtonian model. We observed that the orientation of the cell influences the deformation and the imposed shear rate drives the local shear rate distribution along the particle surface. The integral particle deformation for the non-Newtonian models in the given shear rate regime is higher than that for the Newtonian reference model. Finally, the deformation of the cell surface decreases as the dissipation ratio increases.

Finite Element Analysis of Osteocytes Mechanosensitivity Under Simulated Microgravity

NASA Astrophysics Data System (ADS)

Yang, Xiao; Sun, Lian-Wen; Du, Cheng-Fei; Wu, Xin-Tong; Fan, Yu-Bo

2018-04-01

It was found that the mechanosensitivity of osteocytes could be altered under simulated microgravity. However, how the mechanical stimuli as the biomechanical origins cause the bioresponse in osteocytes under microgravity is unclear yet. Computational studies may help us to explore the mechanical deformation changes of osteocytes under microgravity. Here in this paper, we intend to use the computational simulation to investigate the mechanical behavior of osteocytes under simulated microgravity. In order to obtain the shape information of osteocytes, the biological experiment was conducted under simulated microgravity prior to the numerical simulation The cells were rotated by a clinostat for 6 hours or 5 days and fixed, the cytoskeleton and the nucleus were immunofluorescence stained and scanned, and the cell shape and the fluorescent intensity were measured from fluorescent images to get the dimension information of osteocytes The 3D finite element (FE) cell models were then established based on the scanned image stacks. Several components such as the actin cortex, the cytoplasm, the nucleus, the cytoskeleton of F-actin and microtubules were considered in the model. The cell models in both 6 hours and 5 days groups were then imposed by three magnitudes (0.5, 10 and 15 Pa) of simulating fluid shear stress, with cell total displacement and the internal discrete components deformation calculated. The results showed that under the simulated microgravity: (1) the nuclear area and height statistically significantly increased, which made the ratio of membrane-cortex height to nucleus height statistically significantly decreased; (2) the fluid shear stress-induced maximum displacements and average displacements in the whole cell decreased, with the deformation decreasing amplitude was largest when exposed to 1.5Pa of fluid shear stress; (3) the fluid shear stress-induced deformation of cell membrane-cortex and cytoskeleton decreased, while the fluid shear stress-induced deformation of nucleus increased. The results suggested the mechanical behavior of whole osteocyte cell body was suppressed by simulated microgravity, and this decrement was enlarged with either the increasing amplitude of fluid shear stress or the duration of simulated microgravity. What's more, the mechanical behavior of membrane-cortex and cytoskeleton was suppressed by the simulated microgravity, which indicated the mechanotransduction process in the cell body may be further inhibited. On the contrary, the cell nucleus deformation increased under simulated microgravity, which may be related to either the decreased amount of cytoskeleton or the increased volume occupied proportion of nucleus in whole cell under the simulated microgravity. The numerical results supported our previous biological experiments, and showed particularly affected cellular components under the simulated microgravity. The computational study here may help us to better understand the mechanism of mechanosensitivity changes in osteocytes under simulated microgravity, and further to explore the mechanism of the bone loss in space flight.

Multiscale Modeling of Virus Entry via Receptor-Mediated Endocytosis

NASA Astrophysics Data System (ADS)

Liu, Jin

2012-11-01

Virus infections are ubiquitous and remain major threats to human health worldwide. Viruses are intracellular parasites and must enter host cells to initiate infection. Receptor-mediated endocytosis is the most common entry pathway taken by viruses, the whole process is highly complex and dictated by various events, such as virus motions, membrane deformations, receptor diffusion and ligand-receptor reactions, occurring at multiple length and time scales. We develop a multiscale model for virus entry through receptor-mediated endocytosis. The binding of virus to cell surface is based on a mesoscale three dimensional stochastic adhesion model, the internalization (endocytosis) of virus and cellular membrane deformation is based on the discretization of Helfrich Hamiltonian in a curvilinear space using Monte Carlo method. The multiscale model is based on the combination of these two models. We will implement this model to study the herpes simplex virus entry into B78 cells and compare the model predictions with experimental measurements.

Mechanics of membrane-cytoskeleton attachment in Paramecium

NASA Astrophysics Data System (ADS)

Campillo, C.; Jerber, J.; Fisch, C.; Simoes-Betbeder, M.; Dupuis-Williams, P.; Nassoy, P.; Sykes, C.

2012-12-01

In this paper we assess the role of the protein MKS1 (Meckel syndrome type 1) in the cortical membrane mechanics of the ciliated protist Paramecium. This protein is known to be crucial in the process of cilium formation, and we investigate its putative role in membrane-cytoskeleton attachment. Therefore, we compare cells where the gene coding for MKS1 is silenced to wild-type cells. We found that scanning electron microscopy observation of the cell surface reveals a cup-like structure in wild-type cells that is lost in silenced cells. Since this structure is based on the underlying cytoskeleton, one hypothesis to explain this observation is a disruption of membrane attachment to the cytoskeleton in the absence of MKS1 that should affect plasma membrane mechanics. We test this by probing the mechanics of wild-type and silenced cells by micropipette aspiration. Strikingly, we observe that, at the same aspiration pressure, the membrane of silenced cells is easily aspirated by the micropipette whereas that of wild-type cells enters only at a moderate velocity, an effect that suggests a detachment of the membrane from the underlying cytoskeleton in silenced cells. We quantify this detachment by measuring the deformation of the cell cortex and the rate of cell membrane entry in the micropipette. This study offers a new perspective for the characterization of membrane-cytoskeleton attachment in protists and paves the way for a better understanding of the role of membrane-cortex attachment in cilium formation.

Deformation of phospholipid vesicles in an optical stretcher.

PubMed

Delabre, Ulysse; Feld, Kasper; Crespo, Eleonore; Whyte, Graeme; Sykes, Cecile; Seifert, Udo; Guck, Jochen

2015-08-14

Phospholipid vesicles are common model systems for cell membranes. Important aspects of the membrane function relate to its mechanical properties. Here we have investigated the deformation behaviour of phospholipid vesicles in a dual-beam laser trap, also called an optical stretcher. This study explicitly makes use of the inherent heating present in such traps to investigate the dependence of vesicle deformation on temperature. By using lasers with different wavelengths, optically induced mechanical stresses and temperature increase can be tuned fairly independently with a single setup. The phase transition temperature of vesicles can be clearly identified by an increase in deformation. In the case of no heating effects, a minimal model for drop deformation in an optical stretcher and a more specific model for vesicle deformation that takes explicitly into account the angular dependence of the optical stress are presented to account for the experimental results. Elastic constants are extracted from the fitting procedures, which agree with literature data. This study demonstrates the utility of optical stretching, which is easily combined with microfluidic delivery, for the future serial, high-throughput study of the mechanical and thermodynamic properties of phospholipid vesicles.

External push and internal pull forces recruit curvature sensing N-BAR domain proteins to the plasma membrane

PubMed Central

Galic, Milos; Jeong, Sangmoo; Tsai, Feng-Chiao; Joubert, Lydia-Marie; Wu, Yi I.; Hahn, Klaus M.; Cui, Yi; Meyer, Tobias

2012-01-01

Many of the more than 20 mammalian proteins with N-BAR domains1-2 control cell architecture3 and endocytosis4-5 by associating with curved sections of the plasma membrane (PM)6. It is not well understood whether N-BAR proteins are recruited directly by processes that mechanically curve the PM or indirectly by PM-associated adaptor proteins that recruit proteins with N-BAR domains that then induce membrane curvature. Here, we show that externally-induced inward deformation of the PM by cone-shaped nanostructures (Nanocones) and internally-induced inward deformation by contracting actin cables both trigger recruitment of isolated N-BAR domains to the curved PM. Markedly, live-cell imaging in adherent cells showed selective recruitment of full length N-BAR proteins and isolated N-BAR domains to PM sub-regions above Nanocone stripes. Electron microscopy confirmed that N-BAR domains are recruited to local membrane sites curved by Nanocones. We further showed that N-BAR domains are periodically recruited to curved PM sites during local lamellipodia retraction in the front of migrating cells. Recruitment required Myosin II-generated force applied to PM connected actin cables. Together, our study shows that N-BAR domains can be directly recruited to the PM by external push or internal pull forces that locally curve the PM. PMID:22750946

Coupled elasticity-diffusion model for the effects of cytoskeleton deformation on cellular uptake of cylindrical nanoparticles.

PubMed

Wang, Jizeng; Li, Long

2015-01-06

Molecular dynamic simulations and experiments have recently demonstrated how cylindrical nanoparticles (CNPs) with large aspect ratios penetrate animal cells and inevitably deform cytoskeletons. Thus, a coupled elasticity-diffusion model was adopted to elucidate this interesting biological phenomenon by considering the effects of elastic deformations of cytoskeleton and membrane, ligand-receptor binding and receptor diffusion. The mechanism by which the binding energy drives the CNPs with different orientations to enter host cells was explored. This mechanism involved overcoming the resistance caused by cytoskeleton and membrane deformations and the change in configurational entropy of the ligand-receptor bonds and free receptors. Results showed that deformation of the cytoskeleton significantly influenced the engulfing process by effectively slowing down and even hindering the entry of the CNPs. Additionally, the engulfing depth was determined quantitatively. CNPs preferred or tended to vertically attack target cells until they were stuck in the cytoskeleton as implied by the speed of vertically oriented CNPs that showed much faster initial engulfing speeds than horizontally oriented CNPs. These results elucidated the most recent molecular dynamics simulations and experimental observations on the cellular uptake of carbon nanotubes and phagocytosis of filamentous Escherichia coli bacteria. The most efficient engulfment showed the stiffness-dependent optimal radius of the CNPs. Cytoskeleton stiffness exhibited more significant influence on the optimal sizes of the vertical uptake than the horizontal uptake. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

Guided by curvature: shaping cells by coupling curved membrane proteins and cytoskeletal forces.

PubMed

Gov, N S

2018-05-26

Eukaryote cells have flexible membranes that allow them to have a variety of dynamical shapes. The shapes of the cells serve important biological functions, both for cells within an intact tissue, and during embryogenesis and cellular motility. How cells control their shapes and the structures that they form on their surface has been a subject of intensive biological research, exposing the building blocks that cells use to deform their membranes. These processes have also drawn the interest of theoretical physicists, aiming to develop models based on physics, chemistry and nonlinear dynamics. Such models explore quantitatively different possible mechanisms that the cells can employ to initiate the spontaneous formation of shapes and patterns on their membranes. We review here theoretical work where one such class of mechanisms was investigated: the coupling between curved membrane proteins, and the cytoskeletal forces that they recruit. Theory indicates that this coupling gives rise to a rich variety of membrane shapes and dynamics, while experiments indicate that this mechanism appears to drive many cellular shape changes.This article is part of the theme issue 'Self-organization in cell biology'. © 2018 The Author(s).

Oligomerization but Not Membrane Bending Underlies the Function of Certain F-BAR Proteins in Cell Motility and Cytokinesis.

PubMed

McDonald, Nathan A; Vander Kooi, Craig W; Ohi, Melanie D; Gould, Kathleen L

2015-12-21

F-BAR proteins function in diverse cellular processes by linking membranes to the actin cytoskeleton. Through oligomerization, multiple F-BAR domains can bend membranes into tubules, though the physiological importance of F-BAR-to-F-BAR assemblies is not yet known. Here, we investigate the F-BAR domain of the essential cytokinetic scaffold, Schizosaccharomyces pombe Cdc15, during cytokinesis. Challenging a widely held view that membrane deformation is a fundamental property of F-BARs, we report that the Cdc15 F-BAR binds, but does not deform, membranes in vivo or in vitro, and six human F-BAR domains-including those from Fer and RhoGAP4-share this property. Nevertheless, tip-to-tip interactions between F-BAR dimers are critical for Cdc15 oligomerization and high-avidity membrane binding, stabilization of contractile ring components at the medial cortex, and the fidelity of cytokinesis. F-BAR oligomerization is also critical for Fer and RhoGAP4 physiological function, demonstrating its broad importance to F-BAR proteins that function without membrane bending. Copyright © 2015 Elsevier Inc. All rights reserved.

Computational Modeling and Simulations of Bioparticle Internalization Through Clathrin-mediated Endocytosis

NASA Astrophysics Data System (ADS)

Deng, Hua; Dutta, Prashanta; Liu, Jin

2016-11-01

Clathrin-mediated endocytosis (CME) is one of the most important endocytic pathways for the internalization of bioparticles at lipid membrane of cells, which plays crucial roles in fundamental understanding of viral infections and interacellular/transcelluar targeted drug delivery. During CME, highly dynamic clathrin-coated pit (CCP), formed by the growth of ordered clathrin lattices, is the key scaffolding component that drives the deformation of plasma membrane. Experimental studies have shown that CCP alone can provide sufficient membrane curvature for facilitating membrane invagination. However, currently there is no computational model that could couple cargo receptor binding with membrane invagination process, nor simulations of the dynamic growing process of CCP. We develop a stochastic computational model for the clathrin-mediated endocytosis based on Metropolis Monte Carlo simulations. In our model, the energetic costs of bending membrane and CCP are linked with antigen-antibody interactions. The assembly of clathrin lattices is a dynamic process that correlates with antigen-antibody bond formation. This model helps study the membrane deformation and the effects of CCP during functionalized bioparticles internalization through CME. This work is supported by NSF Grants: CBET-1250107 and CBET-1604211.

Inflight Assay of Red Blood Cell Deformability

NASA Technical Reports Server (NTRS)

Ingram, M.; Paglia, D. E.; Eckstein, E. C.; Frazer, R. E.

1985-01-01

Studies on Soviet and American astronauts have demonstrated that red blood cell production is altered in response to low gravity (g) environment. This is associated with changes in individual red cells including increased mean cell volume and altered membrane deformability. During long orbital missions, there is a tendency for the red cell mass deficit to be at least partly corrected although the cell shape anomalies are not. Data currently available suggest that the observed decrease in red cell mass is the result of sudden suppression of erythropoieses and that the recovery trend observed during long missions reflects re-establishment of erythropoietic homeostasis at a "set point" for the red cell mass that is slightly below the normal level at 1 g.

Shape matters in protein mobility within membranes

PubMed Central

Quemeneur, François; Sigurdsson, Jon K.; Renner, Marianne; Atzberger, Paul J.; Bassereau, Patricia; Lacoste, David

2014-01-01

The lateral mobility of proteins within cell membranes is usually thought to be dependent on their size and modulated by local heterogeneities of the membrane. Experiments using single-particle tracking on reconstituted membranes demonstrate that protein diffusion is significantly influenced by the interplay of membrane curvature, membrane tension, and protein shape. We find that the curvature-coupled voltage-gated potassium channel (KvAP) undergoes a significant increase in protein mobility under tension, whereas the mobility of the curvature-neutral water channel aquaporin 0 (AQP0) is insensitive to it. Such observations are well explained in terms of an effective friction coefficient of the protein induced by the local membrane deformation. PMID:24706877

Red blood cell vesiculation in hereditary hemolytic anemia

PubMed Central

Alaarg, Amr; Schiffelers, Raymond M.; van Solinge, Wouter W.; van Wijk, Richard

2013-01-01

Hereditary hemolytic anemia encompasses a heterogeneous group of anemias characterized by decreased red blood cell survival because of inherited membrane, enzyme, or hemoglobin disorders. Affected red blood cells are more fragile, less deformable, and more susceptible to shear stress and oxidative damage, and show increased vesiculation. Red blood cells, as essentially all cells, constitutively release phospholipid extracellular vesicles in vivo and in vitro in a process known as vesiculation. These extracellular vesicles comprise a heterogeneous group of vesicles of different sizes and intracellular origins. They are described in literature as exosomes if they originate from multi-vesicular bodies, or as microvesicles when formed by a one-step budding process directly from the plasma membrane. Extracellular vesicles contain a multitude of bioactive molecules that are implicated in intercellular communication and in different biological and pathophysiological processes. Mature red blood cells release in principle only microvesicles. In hereditary hemolytic anemias, the underlying molecular defect affects and determines red blood cell vesiculation, resulting in shedding microvesicles of different compositions and concentrations. Despite extensive research into red blood cell biochemistry and physiology, little is known about red cell deformability and vesiculation in hereditary hemolytic anemias, and the associated pathophysiological role is incompletely assessed. In this review, we discuss recent progress in understanding extracellular vesicles biology, with focus on red blood cell vesiculation. Also, we review recent scientific findings on the molecular defects of hereditary hemolytic anemias, and their correlation with red blood cell deformability and vesiculation. Integrating bio-analytical findings on abnormalities of red blood cells and their microvesicles will be critical for a better understanding of the pathophysiology of hereditary hemolytic anemias. PMID:24379786

[Principles of changes of structural organization of cell membranes and functional properties of erythrocytes in neurotic disorders].

PubMed

Riazantseva, N V; Novitskiĭ, V V

2003-02-01

Investigation into structural, metabolic, and functional conditions of red blood cells was performed in 24 patients with a neurosis (neurasthenia, disturbance of asaptation) with the aid of electrophoretic division of proteins of the erythrocyte membrane, thin-layer chromatography, fluorescent probing of membranes, evaluation of peroxidative oxidation process, scanning and transmission electron microscopy, laser diphractometry, photometry. The patients with neurotic disorders at the early period after the influence of psychogenic factors (up to 3 months) revealed disorganization of lipid and protein composition of the red cell membrane, increase in microviscosity of its lipid phase, impairment of surface architectonics and ultrastructure of red cells, decrease of a deformation ability and increase of aggregate properties of erythrocytes. The authors treat stability of erythrocytes' homeostasis under the long-term influence of psychogenic factors from a viewpoint of adaptive changes in organism under the influence of neurogenic factors.

Deformation and relaxation of an incompressible viscoelastic body with surface viscoelasticity

NASA Astrophysics Data System (ADS)

Liu, Liping; Yu, Miao; Lin, Hao; Foty, Ramsey

2017-01-01

Measuring mechanical properties of cells or cell aggregates has proven to be an involved process due to their geometrical and structural complexity. Past measurements are based on material models that completely neglect the elasticity of either the surface membrane or the interior bulk. In this work, we consider general material models to account for both surface and bulk viscoelasticity. The boundary value problems are formulated for deformations and relaxations of a closed viscoelastic surface coupled with viscoelastic media inside and outside of the surface. The linearized surface elasticity models are derived for the constant surface tension model and the Helfrich-Canham bending model for coupling with the bulk viscoelasticity. For quasi-spherical surfaces, explicit solutions are obtained for the deformation, stress-strain and relaxation behaviors under a variety of loading conditions. These solutions can be applied to extract the intrinsic surface and bulk viscoelastic properties of biological cells or cell aggregates in the indentation, electro-deformation and relaxation experiments.

Toward the Structure of Dynamic Membrane-Anchored Actin Networks

PubMed Central

Weber, Igor

2007-01-01

In the cortex of a motile cell, membrane-anchored actin filaments assemble into structures of varying shape and function. Filopodia are distinguished by a core of bundled actin filaments within finger-like extensions of the membrane. In a recent paper by Medalia et al1 cryo-electron tomography has been used to reconstruct, from filopodia of Dictyostelium cells, the 3-dimensional organization of actin filaments in connection with the plasma membrane. A special arrangement of short filaments converging toward the filopod's tip has been called a “terminal cone”. In this region force is applied for protrusion of the membrane. Here we discuss actin organization in the filopodia of Dictyostelium in the light of current views on forces that are generated by polymerizing actin filaments, and on the resistance of membranes against deformation that counteracts these forces. PMID:19262130

Reversibility of red blood cell deformation

NASA Astrophysics Data System (ADS)

Zeitz, Maria; Sens, P.

2012-05-01

The ability of cells to undergo reversible shape changes is often crucial to their survival. For red blood cells (RBCs), irreversible alteration of the cell shape and flexibility often causes anemia. Here we show theoretically that RBCs may react irreversibly to mechanical perturbations because of tensile stress in their cytoskeleton. The transient polymerization of protein fibers inside the cell seen in sickle cell anemia or a transient external force can trigger the formation of a cytoskeleton-free membrane protrusion of μm dimensions. The complex relaxation kinetics of the cell shape is shown to be responsible for selecting the final state once the perturbation is removed, thereby controlling the reversibility of the deformation. In some case, tubular protrusion are expected to relax via a peculiar “pearling instability.”

Reversibility of red blood cell deformation.

PubMed

Zeitz, Maria; Sens, P

2012-05-01

The ability of cells to undergo reversible shape changes is often crucial to their survival. For red blood cells (RBCs), irreversible alteration of the cell shape and flexibility often causes anemia. Here we show theoretically that RBCs may react irreversibly to mechanical perturbations because of tensile stress in their cytoskeleton. The transient polymerization of protein fibers inside the cell seen in sickle cell anemia or a transient external force can trigger the formation of a cytoskeleton-free membrane protrusion of μm dimensions. The complex relaxation kinetics of the cell shape is shown to be responsible for selecting the final state once the perturbation is removed, thereby controlling the reversibility of the deformation. In some case, tubular protrusion are expected to relax via a peculiar "pearling instability."

Mechanics of membrane bulging during cell-wall disruption in Gram-negative bacteria

NASA Astrophysics Data System (ADS)

Daly, Kristopher E.; Huang, Kerwyn Casey; Wingreen, Ned S.; Mukhopadhyay, Ranjan

2011-04-01

The bacterial cell wall is a network of sugar strands crosslinked by peptides that serve as the primary structure for bearing osmotic stress. Despite its importance in cellular survival, the robustness of the cell wall to network defects has been relatively unexplored. Treatment of the Gram-negative bacterium Escherichia coli with the antibiotic vancomycin, which disrupts the crosslinking of new material during growth, leads to the development of pronounced bulges and eventually of cell lysis. Here, we model the mechanics of the bulging of the cytoplasmic membrane through pores in the cell wall. We find that the membrane undergoes a transition between a nearly flat state and a spherical bulge at a critical pore radius of ~20 nm. This critical pore size is large compared to the typical distance between neighboring peptides and glycan strands, and hence pore size acts as a constraint on network integrity. We also discuss the general implications of our model to membrane deformations in eukaryotic blebbing and vesiculation in red blood cells.

Red cell membrane skeleton: structure-function relationships.

PubMed

Palek, J; Liu, S C

1980-01-01

This papaer reviews our present understanding of ultrastructure, organization, and functional characteristics of the erythrocyte membrane cytoskeleton. This two-dimensional fibrillar network of submembrane proteins can be visualized after extraction of lipids and integral membrane proteins by Triton X-100. Current data suggest that the major structural components of the cytoskeleton are heterodimers of double-stranded spectrin that form tetramers by head-to-head associations. The tetramers may be connected into a fibrillar meshwork by oligomers of actin. The control of membrane integrity by this network is illustrated by examples of two hemolyotic anemias characterized by marked membrane instability and vesiculation: 1) hereditary spherocytic anemia of the house mouse associated with spectrin deficiency and 2) hereditary pyropoikilocytosis, a hemolytic anemia in man characterized by thermal instability of the membrane and the presence of abnormal spectrin, which exhibits an increased propensity to thermal denaturation. Stabilization of the cytoskeletal network by covalent cross-links between the nearest cytoskeletal and integral membrane proteins results in a decrease of membrane deformability and a fixation of erythrocytes in their abnormal shape. Such cross-linkings include: 1) transamidative cross-links produced by introduction of Ca2+ (>0.5 mM) into fresh erythrocytes, and 2) intermolecular disulfide couplings, which are formed after extensive oxidation of fresh erythrocytes or after mild oxidation of ATP-depleted, but not fresh, erythrocytes. The significance of these cross-links in stabilization of shape of abnormal erythrocytes such as schistocytes remains to be determined. We conclude that spectrin and actin form a fibrillar submembrane network that plays an important role in control of membrane integrity, erythrocyte deformability, and stabilization of cells in abnormal shapes.

Dynamic modeling for flow-activated chloride-selective membrane current in vascular endothelial cells.

PubMed

Qin, Kai-Rong; Xiang, Cheng; Cao, Ling-Ling

2011-10-01

In this paper, a dynamic model is proposed to quantify the relationship between fluid flow and Cl(-)-selective membrane current in vascular endothelial cells (VECs). It is assumed that the external shear stress would first induce channel deformation in VECs. This deformation could activate the Cl(-) channels on the membrane, thus allowing Cl(-) transport across the membrane. A modified Hodgkin-Huxley model is embedded into our dynamic system to describe the electrophysiological properties of the membrane, such as the Cl(-)-selective membrane current (I), voltage (V) and conductance. Three flow patterns, i. e., steady flow, oscillatory flow, and pulsatile flow, are applied in our simulation studies. When the extracellular Cl(-) concentration is constant, the I-V characteristics predicted by our dynamic model shows strong consistency with the experimental observations. It is also interesting to note that the Cl(-) currents under different flow patterns show some differences, indicating that VECs distinguish among and respond differently to different types of flows. When the extracellular Cl(-) concentration keeps constant or varies slowly with time (i.e. oscillates at 0.02 Hz), the convection and diffusion of Cl(-) in extracellular space can be ignored and the Cl(-) current is well captured by the modified Hodgkin-Huxley model alone. However, when the extracellular Cl(-) varies fast (i.e., oscillates at 0.2 Hz), the convection and diffusion effect should be considered because the Cl(-) current dynamics is different from the case where the convection-diffusion effect is simply ignored. The proposed dynamic model along with the simulation results could not only provide more insights into the flow-regulated electrophysiological behavior of the cell membrane but also help to reveal new findings in the electrophysiological experimental investigations of VECs in response to dynamic flow and biochemical stimuli.

Deformation of an elastic capsule in a uniform electric field

NASA Astrophysics Data System (ADS)

Karyappa, Rahul B.; Deshmukh, Shivraj. D.; Thaokar, Rochish. M.

2014-12-01

The deformation of a thin elastic capsule subjected to a uniform electric field is investigated in the Stokes flow regime. The electrohydrodynamic flow is analyzed using a perfect conductor and a perfect dielectric model for the capsule and the fluid phase, respectively. A theoretical analysis is carried out using an asymptotic expansion in the electric capillary number (Ca) (a ratio of the electric stress to the elastic tension) in the small deformation limit using the finite deformation Hooke's law. The analysis is used to determine the elasticity of polysiloxane capsules suspended in oil, the deformation of which is obtained using videography. The boundary element method is implemented to seek numerical solutions to the hydrodynamic, elastic, and electrostatics equations. The finite deformation Hooke's law, the Mooney-Rivlin, and Skalak's model for elasticity are employed. The effect of electric capillary number, unstressed geometry, and the type of membrane material on the deformation of a capsule is presented in the high Ca number limit using numerical simulation. Capsules synthesized with higher monomer concentration displayed electric stress induced wrinkling process at high electric field strengths. Burst of a capsule is characterized by poration of the polymer membrane, which could be symmetric or asymmetric at the two poles, depending upon the value of the capillary number. The results should be useful in understanding the response of elastic capsules such as red blood cells and polymerized membranes, to an electric field, in applications such as electrodeformation and electroporation. It also provides a theoretical framework for a possible way of determining the elastic parameters of a capsule.

Radiation pressure on a biconcave human Red Blood Cell and the resulting deformation in a pair of parallel optical traps.

PubMed

Liao, Guan-Bo; Chen, Yin-Quan; Bareil, Paul B; Sheng, Yunlong; Chiou, Arthur; Chang, Ming-Shien

2014-10-01

We calculated the three-dimensional optical stress distribution and the resulting deformation on a biconcave human red blood cell (RBC) in a pair of parallel optical trap. We assumed a Gaussian intensity distribution with a spherical wavefront for each trapping beam and calculated the optical stress from the momentum transfer associated with the reflection and refraction of the incident photons at each interface. The RBC was modelled as a biconcave thin elastic membrane with uniform elasticity and a uniform thickness of 0.25 μm. The resulting cell deformation was determined from the optical stress distribution by finite element software, Comsol Structure Mechanics Module, with Young's modulus (E) as a fitting parameter in order to fit the theoretical results for cell elongation to our experimental data. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Structural and mechanical heterogeneity of the erythrocyte membrane reveals hallmarks of membrane stability.

PubMed

Picas, Laura; Rico, Félix; Deforet, Maxime; Scheuring, Simon

2013-02-26

The erythrocyte membrane, a metabolically regulated active structure that comprises lipid molecules, junctional complexes, and the spectrin network, enables the cell to undergo large passive deformations when passing through the microvascular system. Here we use atomic force microscopy (AFM) imaging and quantitative mechanical mapping at nanometer resolution to correlate structure and mechanics of key components of the erythrocyte membrane, crucial for cell integrity and function. Our data reveal structural and mechanical heterogeneity modulated by the metabolic state at unprecedented nanometer resolution. ATP-depletion, reducing skeletal junction phosphorylation in RBC cells, leads to membrane stiffening. Analysis of ghosts and shear-force opened erythrocytes show that, in the absence of cytosolic kinases, spectrin phosphorylation results in membrane stiffening at the extracellular face and a reduced junction remodeling in response to loading forces. Topography and mechanical mapping of single components at the cytoplasmic face reveal that, surprisingly, spectrin phosphorylation by ATP softens individual filaments. Our findings suggest that, besides the mechanical signature of each component, the RBC membrane mechanics is regulated by the metabolic state and the assembly of its structural elements.

Modeling thermal inkjet and cell printing process using modified pseudopotential and thermal lattice Boltzmann methods.

PubMed

Sohrabi, Salman; Liu, Yaling

2018-03-01

Pseudopotential lattice Boltzmann methods (LBMs) can simulate a phase transition in high-density ratio multiphase flow systems. If coupled with thermal LBMs through equation of state, they can be used to study instantaneous phase transition phenomena with a high-temperature gradient where only one set of formulations in an LBM system can handle liquid, vapor, phase transition, and heat transport. However, at lower temperatures an unrealistic spurious current at the interface introduces instability and limits its application in real flow system. In this study, we proposed new modifications to the LBM system to minimize a spurious current which enables us to study nucleation dynamic at room temperature. To demonstrate the capabilities of this approach, the thermal ejection process is modeled as one example of a complex flow system. In an inkjet printer, a thermal pulse instantly heats up the liquid in a microfluidic chamber and nucleates bubble vapor providing the pressure pulse necessary to eject droplets at high speed. Our modified method can present a more realistic model of the explosive vaporization process since it can also capture a high-temperature/density gradient at nucleation region. Thermal inkjet technology has been successfully applied for printing cells, but cells are susceptible to mechanical damage or death as they squeeze out of the nozzle head. To study cell deformation, a spring network model, representing cells, is connected to the LBM through the immersed boundary method. Looking into strain and stress distribution of a cell membrane at its most deformed state, it is found that a high stretching rate effectively increases the rupture tension. In other words, membrane deformation energy is released through creation of multiple smaller nanopores rather than big pores. Overall, concurrently simulating multiphase flow, phase transition, heat transfer, and cell deformation in one unified LB platform, we are able to provide a better insight into the bubble dynamic and cell mechanical damage during the printing process.

Modeling thermal inkjet and cell printing process using modified pseudopotential and thermal lattice Boltzmann methods

NASA Astrophysics Data System (ADS)

Sohrabi, Salman; Liu, Yaling

2018-03-01

Pseudopotential lattice Boltzmann methods (LBMs) can simulate a phase transition in high-density ratio multiphase flow systems. If coupled with thermal LBMs through equation of state, they can be used to study instantaneous phase transition phenomena with a high-temperature gradient where only one set of formulations in an LBM system can handle liquid, vapor, phase transition, and heat transport. However, at lower temperatures an unrealistic spurious current at the interface introduces instability and limits its application in real flow system. In this study, we proposed new modifications to the LBM system to minimize a spurious current which enables us to study nucleation dynamic at room temperature. To demonstrate the capabilities of this approach, the thermal ejection process is modeled as one example of a complex flow system. In an inkjet printer, a thermal pulse instantly heats up the liquid in a microfluidic chamber and nucleates bubble vapor providing the pressure pulse necessary to eject droplets at high speed. Our modified method can present a more realistic model of the explosive vaporization process since it can also capture a high-temperature/density gradient at nucleation region. Thermal inkjet technology has been successfully applied for printing cells, but cells are susceptible to mechanical damage or death as they squeeze out of the nozzle head. To study cell deformation, a spring network model, representing cells, is connected to the LBM through the immersed boundary method. Looking into strain and stress distribution of a cell membrane at its most deformed state, it is found that a high stretching rate effectively increases the rupture tension. In other words, membrane deformation energy is released through creation of multiple smaller nanopores rather than big pores. Overall, concurrently simulating multiphase flow, phase transition, heat transfer, and cell deformation in one unified LB platform, we are able to provide a better insight into the bubble dynamic and cell mechanical damage during the printing process.

Actin protofilament orientation in deformation of the erythrocyte membrane skeleton.

PubMed Central

Picart, C; Dalhaimer, P; Discher, D E

2000-01-01

The red cell's spectrin-actin network is known to sustain local states of shear, dilation, and condensation, and yet the short actin filaments are found to maintain membrane-tangent and near-random azimuthal orientations. When calibrated with polarization results for single actin filaments, imaging of micropipette-deformed red cell ghosts has allowed an assessment of actin orientations and possible reorientations in the network. At the hemispherical cap of the aspirated projection, where the network can be dilated severalfold, filaments have the same membrane-tangent orientation as on a relatively unstrained portion of membrane. Likewise, over the length of the network projection pulled into the micropipette, where the network is strongly sheared in axial extension and circumferential contraction, actin maintains its tangent orientation and is only very weakly aligned with network extension. Similar results are found for the integral membrane protein Band 3. Allowing for thermal fluctuations, we deduce a bound for the effective coupling constant, alpha, between network shear and azimuthal orientation of the protofilament. The finding that alpha must be about an order of magnitude or more below its tight-coupling value illustrates how nanostructural kinematics can decouple from more macroscopic responses. Monte Carlo simulations of spectrin-actin networks at approximately 10-nm resolution further support this conclusion and substantiate an image of protofilaments as elements of a high-temperature spin glass. PMID:11106606

Arenavirus budding resulting from viral-protein-associated cell membrane curvature

PubMed Central

Schley, David; Whittaker, Robert J.; Neuman, Benjamin W.

2013-01-01

Viral replication occurs within cells, with release (and onward infection) primarily achieved through two alternative mechanisms: lysis, in which virions emerge as the infected cell dies and bursts open; or budding, in which virions emerge gradually from a still living cell by appropriating a small part of the cell membrane. Virus budding is a poorly understood process that challenges current models of vesicle formation. Here, a plausible mechanism for arenavirus budding is presented, building on recent evidence that viral proteins embed in the inner lipid layer of the cell membrane. Experimental results confirm that viral protein is associated with increased membrane curvature, whereas a mathematical model is used to show that localized increases in curvature alone are sufficient to generate viral buds. The magnitude of the protein-induced curvature is calculated from the size of the amphipathic region hypothetically removed from the inner membrane as a result of translation, with a change in membrane stiffness estimated from observed differences in virion deformation as a result of protein depletion. Numerical results are based on experimental data and estimates for three arenaviruses, but the mechanisms described are more broadly applicable. The hypothesized mechanism is shown to be sufficient to generate spontaneous budding that matches well both qualitatively and quantitatively with experimental observations. PMID:23864502

Physical basis of some membrane shaping mechanisms

PubMed Central

2016-01-01

In vesicular transport pathways, membrane proteins and lipids are internalized, externalized or transported within cells, not by bulk diffusion of single molecules, but embedded in the membrane of small vesicles or thin tubules. The formation of these ‘transport carriers’ follows sequential events: membrane bending, fission from the donor compartment, transport and eventually fusion with the acceptor membrane. A similar sequence is involved during the internalization of drug or gene carriers inside cells. These membrane-shaping events are generally mediated by proteins binding to membranes. The mechanisms behind these biological processes are actively studied both in the context of cell biology and biophysics. Bin/amphiphysin/Rvs (BAR) domain proteins are ideally suited for illustrating how simple soft matter principles can account for membrane deformation by proteins. We review here some experimental methods and corresponding theoretical models to measure how these proteins affect the mechanics and the shape of membranes. In more detail, we show how an experimental method employing optical tweezers to pull a tube from a giant vesicle may give important quantitative insights into the mechanism by which proteins sense and generate membrane curvature and the mechanism of membrane scission. This article is part of the themed issue ‘Soft interfacial materials: from fundamentals to formulation’. PMID:27298443

pH sensing by lipids in membranes: The fundamentals of pH-driven migration, polarization and deformations of lipid bilayer assemblies.

PubMed

Angelova, Miglena I; Bitbol, Anne-Florence; Seigneuret, Michel; Staneva, Galya; Kodama, Atsuji; Sakuma, Yuka; Kawakatsu, Toshihiro; Imai, Masayuki; Puff, Nicolas

2018-03-06

Most biological molecules contain acido-basic groups that modulate their structure and interactions. A consequence is that pH gradients, local heterogeneities and dynamic variations are used by cells and organisms to drive or regulate specific biological functions including energetic metabolism, vesicular traffic, migration and spatial patterning of tissues in development. While the direct or regulatory role of pH in protein function is well documented, the role of hydrogen and hydroxyl ions in modulating the properties of lipid assemblies such as bilayer membranes is only beginning to be understood. Here, we review approaches using artificial lipid vesicles that have been instrumental in providing an understanding of the influence of pH gradients and local variations on membrane vectorial motional processes: migration, membrane curvature effects promoting global or local deformations, crowding generation by segregative polarization processes. In the case of pH induced local deformations, an extensive theoretical framework is given and an application to a specific biological issue, namely the structure and stability of mitochondrial cristae, is described. This article is part of a Special Issue entitled: Emergence of Complex Behavior in Biomembranes edited by Marjorie Longo. Copyright © 2018 Elsevier B.V. All rights reserved.

Modeling the Mechanics of Cell Division: Influence of Spontaneous Membrane Curvature, Surface Tension, and Osmotic Pressure

PubMed Central

Beltrán-Heredia, Elena; Almendro-Vedia, Víctor G.; Monroy, Francisco; Cao, Francisco J.

2017-01-01

Many cell division processes have been conserved throughout evolution and are being revealed by studies on model organisms such as bacteria, yeasts, and protozoa. Cellular membrane constriction is one of these processes, observed almost universally during cell division. It happens similarly in all organisms through a mechanical pathway synchronized with the sequence of cytokinetic events in the cell interior. Arguably, such a mechanical process is mastered by the coordinated action of a constriction machinery fueled by biochemical energy in conjunction with the passive mechanics of the cellular membrane. Independently of the details of the constriction engine, the membrane component responds against deformation by minimizing the elastic energy at every constriction state following a pathway still unknown. In this paper, we address a theoretical study of the mechanics of membrane constriction in a simplified model that describes a homogeneous membrane vesicle in the regime where mechanical work due to osmotic pressure, surface tension, and bending energy are comparable. We develop a general method to find approximate analytical expressions for the main descriptors of a symmetrically constricted vesicle. Analytical solutions are obtained by combining a perturbative expansion for small deformations with a variational approach that was previously demonstrated valid at the reference state of an initially spherical vesicle at isotonic conditions. The analytic approximate results are compared with the exact solution obtained from numerical computations, getting a good agreement for all the computed quantities (energy, area, volume, constriction force). We analyze the effects of the spontaneous curvature, the surface tension and the osmotic pressure in these quantities, focusing especially on the constriction force. The more favorable conditions for vesicle constriction are determined, obtaining that smaller constriction forces are required for positive spontaneous curvatures, low or negative membrane tension and hypertonic media. Conditions for spontaneous constriction at a given constriction force are also determined. The implications of these results for biological cell division are discussed. This work contributes to a better quantitative understanding of the mechanical pathway of cellular division, and could assist the design of artificial divisomes in vesicle-based self-actuated microsystems obtained from synthetic biology approaches. PMID:28579960

Interaction of nanoparticles with lipid layers

NASA Astrophysics Data System (ADS)

Park, Jonghyun; Lu, Wei

2009-08-01

Poly (amidoamine) dendrimer nanoparticles are used extensively in diverse biological and medical applications. Examples include gene and drug delivery, where nanoparticles disrupt cell membranes to allow the transport of material into cells. The size and surface chemistry of these particles have a strong effect on their interaction with membranes. This paper proposes a three-dimensional phase-field model to investigate how the interaction drives deformation and morphological evolution of the membrane. Attention is focused on the hole-formation process in the membrane. The simulations have demonstrated that a larger amine-terminated generation 7 dendrimer, which has positive charges, causes the formation of a hole in the membrane. The displaced membrane molecules enclose the particle and form a dendrimer-filled membrane vesicle. The effect is significantly reduced for a smaller dendrimer. An acetamide-terminated dendrimer, which has a neutral charge at the surface, does not cause hole formation. These results agree with experimental observations from atomic force microscopy. The study will provide insight into the design of appropriate nanoparticle surface properties for medical applications.

Biomechanics and dynamics of red blood cells probed by optical tweezers and digital holographic microscopy

NASA Astrophysics Data System (ADS)

Cardenas, Nelson; Thomas, Pattrick; Yu, Lingfeng; Mohanty, Samarendra

2011-03-01

Red blood cells (RBC), with their unique viscoelastic properties, can undergo large deformations during interaction with fluid flow and migration through narrow capillaries. Both local and overall viscoelastic property is important for cellular function and change in these properties indicate diseased condition. Though biomechanics of the cells have been studied using variety of physical techniques (AFM, optically-trapped anchoring beads and microcapilary aspiration) in force regime 10pN, little is studied at low force regime <1pN. Such perturbations are not only hard to exercise on the cell membrane, but quantification of such deformations becomes extremely difficult. By application of low power optical tweezers directly on cell membrane, we could locally perturb discotic RBC along the axial direction, which was monitored dynamically by digital holographic microscopy-a real time, wide-field imaging method having nm axial resolution. The viscoelastic property of the RBC at low force regime was found to be significantly different from that of high-force regime. The results were found to be in good agreement with the simulation results obtained using finite element model of the axially-stretched RBC. The simulations and results of viscoelestic measurements will be presented.

Apparatus for measuring the finite load-deformation behavior of a sheet of epithelial cells cultured on a mesoscopic freestanding elastomer membrane.

PubMed

Selby, John C; Shannon, Mark A

2007-09-01

Details are given for the design, calibration, and operation of an apparatus for measuring the finite load-deformation behavior of a sheet of living epithelial cells cultured on a mesoscopic freestanding elastomer membrane, 10 microm thick and 5 mm in diameter. Although similar in concept to bulge tests used to investigate the mechanical properties of micromachined thin films, cell-elastomer composite diaphragm inflation tests pose a unique set of experimental challenges. Composite diaphragm (CD) specimens are extremely compliant (E<50 kPa), experience large displacements when subject to small inflation pressures (approximately 100 Pa), and must be continuously immersed in a bath of liquid culture medium during the acquisition of load-deformation measurements. Given these considerations, we have constructed an inflation apparatus consisting of an air-piston-cylinder pump integrated with a modular specimen mounting fixture that constitutes a horizontally semi-infinite reservoir of liquid culture medium. In a deformation-controlled inflation test, pressurized air is used to inflate a CD specimen into the liquid reservoir with minimum disturbance of the liquid-air interface. Piston displacements and absolute pump chamber air pressures are utilized as feedback to cycle the displaced (or inflated) CD volume V in a 0.05 Hz triangular or sinusoidal wave form (V(MIN)=0 microl, V(MAX)

Cytoplasmic Fungal Lipases Release Fungicides from Ultra-Deformable Vesicular Drug Carriers

PubMed Central

Steinberg, Gero

2012-01-01

The Transfersome® is a lipid vesicle that contains membrane softeners, such as Tween 80, to make it ultra-deformable. This feature makes the Transfersome® an efficient carrier for delivery of therapeutic drugs across the skin barrier. It was reported that TDT 067 (a topical formulation of 15 mg/ml terbinafine in Transfersome® vesicles) has a much more potent antifungal activity in vitro compared with conventional terbinafine, which is a water-insoluble fungicide. Here we use ultra-structural studies and live imaging in a model fungus to describe the underlying mode of action. We show that terbinafine causes local collapse of the fungal endoplasmic reticulum, which was more efficient when terbinafine was delivered in Transfersome® vesicles (TFVs). When applied in liquid culture, fluorescently labeled TFVs rapidly entered the fungal cells (T1/2∼2 min). Entry was F-actin- and ATP-independent, indicating that it is a passive process. Ultra-structural studies showed that passage through the cell wall involves significant deformation of the vesicles, and depends on a high concentration of the surfactant Tween 80 in their membrane. Surprisingly, the TFVs collapsed into lipid droplets after entry into the cell and the terbinafine was released from their interior. With time, the lipid bodies were metabolized in an ATP-dependent fashion, suggesting that cytosolic lipases attack and degrade intruding TFVs. Indeed, the specific monoacylglycerol lipase inhibitor URB602 prevented Transfersome® degradation and neutralized the cytotoxic effect of Transfersome®-delivered terbinafine. These data suggest that (a) Transfersomes deliver the lipophilic fungicide Terbinafine to the fungal cell wall, (b) the membrane softener Tween 80 allows the passage of the Transfersomes into the fungal cell, and (c) fungal lipases digest the invading Transfersome® vesicles thereby releasing their cytotoxic content. As this mode of action of Transfersomes is independent of the drug cargo, these results demonstrate the potential of Transfersomes in the treatment of all fungal diseases. PMID:22666476

Diagnostic tool for red blood cell membrane disorders: Assessment of a new generation ektacytometer☆

PubMed Central

Da Costa, Lydie; Suner, Ludovic; Galimand, Julie; Bonnel, Amandine; Pascreau, Tiffany; Couque, Nathalie; Fenneteau, Odile; Mohandas, Narla

2016-01-01

Inherited red blood cell (RBC) membrane disorders, such as hereditary spherocytosis, elliptocytosis and hereditary ovalocytosis, result from mutations in genes encoding various RBC membrane and skeletal proteins. The RBC membrane, a composite structure composed of a lipid bilayer linked to a spectrin/actin-based membrane skeleton, confers upon the RBC unique features of deformability and mechanical stability. The disease severity is primarily dependent on the extent of membrane surface area loss. RBC membrane disorders can be readily diagnosed by various laboratory approaches that include RBC cytology, flow cytometry, ektacytometry, electrophoresis of RBC membrane proteins and genetics. The reference technique for diagnosis of RBC membrane disorders is the osmotic gradient ektacytometry. However, in spite of its recognition as the reference technique, this technique is rarely used as a routine diagnosis tool for RBC membrane disorders due to its limited availability. This may soon change as a new generation of ektacytometer has been recently engineered. In this review, we describe the workflow of the samples shipped to our Hematology laboratory for RBC membrane disorder analysis and the data obtained for a large cohort of French patients presenting with RBC membrane disorders using a newly available version of the ektacytomer. PMID:26603718

Mesoscale computational studies of membrane bilayer remodeling by curvature-inducing proteins

PubMed Central

Ramakrishnan, N.; Sunil Kumar, P. B.; Radhakrishnan, Ravi

2014-01-01

Biological membranes constitute boundaries of cells and cell organelles. These membranes are soft fluid interfaces whose thermodynamic states are dictated by bending moduli, induced curvature fields, and thermal fluctuations. Recently, there has been a flood of experimental evidence highlighting active roles for these structures in many cellular processes ranging from trafficking of cargo to cell motility. It is believed that the local membrane curvature, which is continuously altered due to its interactions with myriad proteins and other macromolecules attached to its surface, holds the key to the emergent functionality in these cellular processes. Mechanisms at the atomic scale are dictated by protein-lipid interaction strength, lipid composition, lipid distribution in the vicinity of the protein, shape and amino acid composition of the protein, and its amino acid contents. The specificity of molecular interactions together with the cooperativity of multiple proteins induce and stabilize complex membrane shapes at the mesoscale. These shapes span a wide spectrum ranging from the spherical plasma membrane to the complex cisternae of the Golgi apparatus. Mapping the relation between the protein-induced deformations at the molecular scale and the resulting mesoscale morphologies is key to bridging cellular experiments across the various length scales. In this review, we focus on the theoretical and computational methods used to understand the phenomenology underlying protein-driven membrane remodeling. Interactions at the molecular scale can be computationally probed by all atom and coarse grained molecular dynamics (MD, CGMD), as well as dissipative particle dynamics (DPD) simulations, which we only describe in passing. We choose to focus on several continuum approaches extending the Canham - Helfrich elastic energy model for membranes to include the effect of curvature-inducing proteins and explore the conformational phase space of such systems. In this description, the protein is expressed in the form of a spontaneous curvature field. The approaches include field theoretical methods limited to the small deformation regime, triangulated surfaces and particle-based computational models to investigate the large-deformation regimes observed in the natural state of many biological membranes. Applications of these methods to understand the properties of biological membranes in homogeneous and inhomogeneous environments of proteins, whose underlying curvature fields are either isotropic or anisotropic, are discussed. The diversity in the curvature fields elicits a rich variety of morphological states, including tubes, discs, branched tubes, and caveola. Mapping the thermodynamic stability of these states as a function of tuning parameters such as concentration and strength of curvature induction of the proteins is discussed. The relative stabilities of these self-organized shapes are examined through free-energy calculations. The suite of methods discussed here can be tailored to applications in specific cellular settings such as endocytosis during cargo trafficking and tubulation of filopodial structures in migrating cells, which makes these methods a powerful complement to experimental studies. PMID:25484487

Mesoscale computational studies of membrane bilayer remodeling by curvature-inducing proteins.

PubMed

Ramakrishnan, N; Sunil Kumar, P B; Radhakrishnan, Ravi

2014-10-01

Biological membranes constitute boundaries of cells and cell organelles. These membranes are soft fluid interfaces whose thermodynamic states are dictated by bending moduli, induced curvature fields, and thermal fluctuations. Recently, there has been a flood of experimental evidence highlighting active roles for these structures in many cellular processes ranging from trafficking of cargo to cell motility. It is believed that the local membrane curvature, which is continuously altered due to its interactions with myriad proteins and other macromolecules attached to its surface, holds the key to the emergent functionality in these cellular processes. Mechanisms at the atomic scale are dictated by protein-lipid interaction strength, lipid composition, lipid distribution in the vicinity of the protein, shape and amino acid composition of the protein, and its amino acid contents. The specificity of molecular interactions together with the cooperativity of multiple proteins induce and stabilize complex membrane shapes at the mesoscale. These shapes span a wide spectrum ranging from the spherical plasma membrane to the complex cisternae of the Golgi apparatus. Mapping the relation between the protein-induced deformations at the molecular scale and the resulting mesoscale morphologies is key to bridging cellular experiments across the various length scales. In this review, we focus on the theoretical and computational methods used to understand the phenomenology underlying protein-driven membrane remodeling. Interactions at the molecular scale can be computationally probed by all atom and coarse grained molecular dynamics (MD, CGMD), as well as dissipative particle dynamics (DPD) simulations, which we only describe in passing. We choose to focus on several continuum approaches extending the Canham - Helfrich elastic energy model for membranes to include the effect of curvature-inducing proteins and explore the conformational phase space of such systems. In this description, the protein is expressed in the form of a spontaneous curvature field. The approaches include field theoretical methods limited to the small deformation regime, triangulated surfaces and particle-based computational models to investigate the large-deformation regimes observed in the natural state of many biological membranes. Applications of these methods to understand the properties of biological membranes in homogeneous and inhomogeneous environments of proteins, whose underlying curvature fields are either isotropic or anisotropic, are discussed. The diversity in the curvature fields elicits a rich variety of morphological states, including tubes, discs, branched tubes, and caveola. Mapping the thermodynamic stability of these states as a function of tuning parameters such as concentration and strength of curvature induction of the proteins is discussed. The relative stabilities of these self-organized shapes are examined through free-energy calculations. The suite of methods discussed here can be tailored to applications in specific cellular settings such as endocytosis during cargo trafficking and tubulation of filopodial structures in migrating cells, which makes these methods a powerful complement to experimental studies.

Mesoscale computational studies of membrane bilayer remodeling by curvature-inducing proteins

NASA Astrophysics Data System (ADS)

Ramakrishnan, N.; Sunil Kumar, P. B.; Radhakrishnan, Ravi

2014-10-01

Biological membranes constitute boundaries of cells and cell organelles. These membranes are soft fluid interfaces whose thermodynamic states are dictated by bending moduli, induced curvature fields, and thermal fluctuations. Recently, there has been a flood of experimental evidence highlighting active roles for these structures in many cellular processes ranging from trafficking of cargo to cell motility. It is believed that the local membrane curvature, which is continuously altered due to its interactions with myriad proteins and other macromolecules attached to its surface, holds the key to the emergent functionality in these cellular processes. Mechanisms at the atomic scale are dictated by protein-lipid interaction strength, lipid composition, lipid distribution in the vicinity of the protein, shape and amino acid composition of the protein, and its amino acid contents. The specificity of molecular interactions together with the cooperativity of multiple proteins induce and stabilize complex membrane shapes at the mesoscale. These shapes span a wide spectrum ranging from the spherical plasma membrane to the complex cisternae of the Golgi apparatus. Mapping the relation between the protein-induced deformations at the molecular scale and the resulting mesoscale morphologies is key to bridging cellular experiments across various length scales. In this review, we focus on the theoretical and computational methods used to understand the phenomenology underlying protein-driven membrane remodeling. Interactions at the molecular scale can be computationally probed by all atom and coarse grained molecular dynamics (MD, CGMD), as well as dissipative particle dynamics (DPD) simulations, which we only describe in passing. We choose to focus on several continuum approaches extending the Canham-Helfrich elastic energy model for membranes to include the effect of curvature-inducing proteins and explore the conformational phase space of such systems. In this description, the protein is expressed in the form of a spontaneous curvature field. The approaches include field theoretical methods limited to the small deformation regime, triangulated surfaces and particle-based computational models to investigate the large-deformation regimes observed in the natural state of many biological membranes. Applications of these methods to understand the properties of biological membranes in homogeneous and inhomogeneous environments of proteins, whose underlying curvature fields are either isotropic or anisotropic, are discussed. The diversity in the curvature fields elicits a rich variety of morphological states, including tubes, discs, branched tubes, and caveola. Mapping the thermodynamic stability of these states as a function of tuning parameters such as concentration and strength of curvature induction of the proteins is discussed. The relative stabilities of these self-organized shapes are examined through free-energy calculations. The suite of methods discussed here can be tailored to applications in specific cellular settings such as endocytosis during cargo trafficking and tubulation of filopodial structures in migrating cells, which makes these methods a powerful complement to experimental studies.

Mechanism of erythrocyte death in human population exposed to arsenic through drinking water

DOE Office of Scientific and Technical Information (OSTI.GOV)

Biswas, Debabrata; Banerjee, Mayukh; Sen, Gargi

2008-07-01

Arsenic contamination in drinking water is one of the biggest natural calamities, which has become an imperative threat to human health throughout the world. Abbreviation of erythrocyte lifespan leading to the development of anemia is a common sequel in arsenic exposed population. This study was undertaken to explore the mechanism of cell death in human erythrocytes during chronic arsenic exposure. Results revealed transformation of smooth discoid red cells into evaginated echinocytic form in the exposed individuals. Further distortion converted reversible echinocytes to irreversible spheroechinocytes. Arsenic toxicity increased membrane microviscosity along with an elevation of cholesterol/phospholipid ratio, which hampered the flexibilitymore » of red cell membrane and made them less deformable. Significant increase in the binding of merocyanine 540 with erythrocyte membrane due to arsenic exposure indicated disruption of lipid packing in the outer leaflet of the cell membrane resulting from altered transbilayer phospholipid asymmetry. Arsenic induced eryptosis was characterized by cell shrinkage and exposure of phosphatidylserine at the cell surface. Furthermore, metabolic starvation with depletion of cellular ATP triggered apoptotic removal of erythrocytes from circulation. Significant decrease in reduced glutathione content indicating defective antioxidant capacity was coupled with enhancement of malondialdehyde and protein carbonyl levels, which pointed to oxidative damage to erythrocyte membrane. Arsenic toxicity intervened into red cell membrane integrity eventually leading to membrane destabilization and hemoglobin release. The study depicted the involvement of both erythrophagocytosis and hemolysis in the destruction of human erythrocytes during chronic arsenic exposure.« less

What do we know about mechanical strain in lung alveoli?

PubMed Central

Roan, Esra

2011-01-01

The pulmonary alveolus, terminal gas-exchange unit of the lung, is composed of alveolar epithelial and endothelial cells separated by a thin basement membrane and interstitial space. These cells participate in the maintenance of a delicate system regulated not only by biological factors but also by the mechanical environment of the lung, which undergoes dynamic deformation during breathing. Clinical and animal studies as well as cell culture studies point toward a strong influence of mechanical forces on lung cells and tissues including effects on growth and repair, surfactant release, injury, and inflammation. However, despite substantial advances in our understanding of lung mechanics over the last half century, there are still many unanswered questions regarding the micromechanics of the alveolus and how it deforms during lung inflation. Therefore, the aims of this review are to draw a multidisciplinary account of the mechanics of the alveolus on the basis of its structure, biology, and chemistry and to compare estimates of alveolar deformation from previous studies. PMID:21873445

Cell mechanics: a dialogue.

PubMed

Tao, Jiaxiang; Li, Yizeng; Vig, Dhruv K; Sun, Sean X

2017-03-01

Under the microscope, eukaryotic animal cells can adopt a variety of different shapes and sizes. These cells also move and deform, and the physical mechanisms driving these movements and shape changes are important in fundamental cell biology, tissue mechanics, as well as disease biology. This article reviews some of the basic mechanical concepts in cells, emphasizing continuum mechanics description of cytoskeletal networks and hydrodynamic flows across the cell membrane. We discuss how cells can generate movement and shape changes by controlling mass fluxes at the cell boundary. These mass fluxes can come from polymerization/depolymerization of actin cytoskeleton, as well as osmotic and hydraulic pressure-driven flow of water across the cell membrane. By combining hydraulic pressure control with force balance conditions at the cell surface, we discuss a quantitative mechanism of cell shape and volume control. The broad consequences of this model on cell mechanosensation and tissue mechanics are outlined.

Cell mechanics: a dialogue

PubMed Central

Tao, Jiaxiang; Li, Yizeng; Vig, Dhruv K; Sun, Sean X

2017-01-01

Under the microscope, eukaryotic animal cells can adopt a variety of different shapes and sizes. These cells also move and deform, and the physical mechanisms driving these movements and shape changes are important in fundamental cell biology, tissue mechanics, as well as disease biology. This article reviews some of the basic mechanical concepts in cells, emphasizing continuum mechanics description of cytoskeletal networks and hydrodynamic flows across the cell membrane. We discuss how cells can generate movement and shape changes by controlling mass fluxes at the cell boundary. These mass fluxes can come from polymerization/depolymerization of actin cytoskeleton, as well as osmotic and hydraulic pressure-driven flow of water across the cell membrane. By combining hydraulic pressure control with force balance conditions at the cell surface, we discuss a quantitative mechanism of cell shape and volume control. The broad consequences of this model on cell mechanosensation and tissue mechanics are outlined. PMID:28129208

Cell mechanics: a dialogue

NASA Astrophysics Data System (ADS)

Tao, Jiaxiang; Li, Yizeng; Vig, Dhruv K.; Sun, Sean X.

2017-03-01

Under the microscope, eukaryotic animal cells can adopt a variety of different shapes and sizes. These cells also move and deform, and the physical mechanisms driving these movements and shape changes are important in fundamental cell biology, tissue mechanics, as well as disease biology. This article reviews some of the basic mechanical concepts in cells, emphasizing continuum mechanics description of cytoskeletal networks and hydrodynamic flows across the cell membrane. We discuss how cells can generate movement and shape changes by controlling mass fluxes at the cell boundary. These mass fluxes can come from polymerization/depolymerization of actin cytoskeleton, as well as osmotic and hydraulic pressure-driven flow of water across the cell membrane. By combining hydraulic pressure control with force balance conditions at the cell surface, we discuss a quantitative mechanism of cell shape and volume control. The broad consequences of this model on cell mechanosensation and tissue mechanics are outlined.

Cholesterol segregates into submicrometric domains at the living erythrocyte membrane: evidence and regulation.

PubMed

Carquin, Mélanie; Conrard, Louise; Pollet, Hélène; Van Der Smissen, Patrick; Cominelli, Antoine; Veiga-da-Cunha, Maria; Courtoy, Pierre J; Tyteca, Donatienne

2015-12-01

Although cholesterol is essential for membrane fluidity and deformability, the level of its lateral heterogeneity at the plasma membrane of living cells is poorly understood due to lack of appropriate probe. We here report on the usefulness of the D4 fragment of Clostridium perfringens toxin fused to mCherry (theta*), as specific, non-toxic, sensitive and quantitative cholesterol-labeling tool, using erythrocyte flat membrane. By confocal microscopy, theta* labels cholesterol-enriched submicrometric domains in coverslip-spread but also gel-suspended (non-stretched) fresh erythrocytes, suggesting in vivo relevance. Cholesterol domains on spread erythrocytes are stable in time and space, restricted by membrane:spectrin anchorage via 4.1R complexes, and depend on temperature and sphingomyelin, indicating combined regulation by extrinsic membrane:cytoskeleton interaction and by intrinsic lipid packing. Cholesterol domains partially co-localize with BODIPY-sphingomyelin-enriched domains. In conclusion, we show that theta* is a useful vital probe to study cholesterol organization and demonstrate that cholesterol forms submicrometric domains in living cells.

Microstructure based hygromechanical modelling of deformation of fruit tissue

NASA Astrophysics Data System (ADS)

Abera, M. K.; Wang, Z.; Verboven, P.; Nicolai, B.

2017-10-01

Quality parameters such as firmness and susceptibility to mechanical damage are affected by the mechanical properties of fruit tissue. Fruit tissue is composed of turgid cells that keep cell walls under tension, and intercellular gas spaces where cell walls of neighboring cells have separated. How the structure and properties of these complex microstructures are affecting tissue mechanics is difficult to unravel experimentally. In this contribution, a modelling methodology is presented to calculate the deformation of apple fruit tissue affected by differences in structure and properties of cells and cell walls. The model can be used to perform compression experiments in silico using a hygromechanical model that computes the stress development and water loss during tissue deformation, much like in an actual compression test. The advantage of the model is that properties and structure can be changed to test the influence on the mechanical deformation process. The effect of microstructure, turgor pressure, cell membrane permeability, wall thickness and damping) on the compressibility of the tissue was simulated. Increasing the turgor pressure and thickness of the cell walls results in increased compression resistance of apple tissue increases, as do decreasing cell size and porosity. Geometric variability of the microstructure of tissues plays a major role, affecting results more than other model parameters. Different fruit cultivars were compared, and it was demonstrated, that microstructure variations within a cultivar are so large that interpretation of cultivar-specific effects is difficult.

Effect of cold deformation on the electrochemical behaviour of 304L stainless steel in contaminated sulfuric acid environment

NASA Astrophysics Data System (ADS)

Luo, Hong; Su, Huaizhi; Ying, Guobing; Dong, Chaofang; Li, Xiaogang

2017-12-01

The effect of cold deformation on the microstructure and electrochemical corrosion behaviour of 304L stainless steel in contaminated sulfuric acid solutions (simulated proton exchange membrane fuel cells environments) were evaluated using electron backscatter diffraction analyses, electrochemical measurements, and surface analyses. The internal microstructure,including the grain sizes, angles of the grain boundaries, low coincidence site lattice boundaries, and phase transformations, was changed due to the cold deformation. No noticeable modifications of the pitting corrosion potential were observed during the various deformations, except for a slight enhancement in the passive current density with an increase in the deformation. The CrO3 and metal Ni species in the passive film were investigated after deformation. After heavy deformation (greater than 60%), nickel oxides were detected. Moreover, the Cr/Fe and O2-/OH- ratios in the passive film were higher before deformation, and they decreased with an increase in the deformation level.

DPD simulation on the dynamics of a healthy and infected red blood cell in flow through a constricted channel

NASA Astrophysics Data System (ADS)

Hoque, Sazid Zamal; Anand, D. Vijay; Patnaik, B. S. V.

2017-11-01

The state of the red blood cell (either healthy or infected RBC) will influence its deformation dynamics. Since the pathological condition related to RBC, primarily originates from a single cell infection, therefore, it is important to relate the deformation dynamics to the mechanical properties (such as, bending rigidity and membrane elasticity). In the present study, numerical simulation of a healthy and malaria infected RBC in a constricted channel is analyzed. The flow simulations are carried out using finite sized dissipative particle dynamics (FDPD) method in conjunction with a discrete model that represents the membrane of the RBC. The numerical equivalent of optical tweezers test is validated against the experimental studies. Two different types of constrictions, viz., a converging-diverging type tapered channel and a stenosed microchannel are considered for the simulation. The effect of degree of constriction and the flow rate effect on the RBC is investigated. It was observed that, as the flow rate decreases, the infected RBC completely blocks the micro vessel. The transit time for infected cell drastically increases compared to healthy RBC. Our simulations indicate that, there is a critical flow rate below which infected RBC cannot pass through the micro capillary.

Liquid Droplets Act as "Compass Needles" for the Stresses in a Deformable Membrane.

PubMed

Schulman, Rafael D; Ledesma-Alonso, René; Salez, Thomas; Raphaël, Elie; Dalnoki-Veress, Kari

2017-05-12

We examine the shape of droplets atop deformable thin elastomeric films prepared with an anisotropic tension. As the droplets generate a deformation in the taut film through capillary forces, they assume a shape that is elongated along the high tension direction. By measuring the contact line profile, the tension in the membrane can be completely determined. Minimal theoretical arguments lead to predictions for the droplet shape and membrane deformation that are in excellent agreement with the data. On the whole, the results demonstrate that droplets can be used as probes to map out the stress field in a membrane.

[Membrane model of the regulation of proliferation: the theory and interpretation of an experiment].

PubMed

Volkov, E I

1983-04-01

The role of cell surface physical organization in the cell cycle regulation is analyzed within the framework of the earlier proposed theory (Chernavskii et al., 1982). Two models of cell surface are considered: hard-frame fluid-mosaic model (latticemosaic) and the fluid-mosaic one. The former deals with normal cells. The existence of integral carcasse or "frame" which is formed by the essential part of cross-linked membrane components and may have at least two different conformational states is hypothesized. The second model describes membranes of tumour cells. With the latter theory any mitogen (excluding the restoration of nutrient depletion) reduces the mechanical tensile strength of the frame and stimulates the general structural rearrangement of the plasma membrane. There are only two conformational transitions during the cell cycle which serve as signals for the beginning of S and M phases. If the values of tensile strength are great enough and therefore the conformational transitions are impossible, the cells pass into the resting (prereplicative--G01, or premitotical--G02) state. Three types of experiments are interpreted in the proposed theory: a) on differences in the action of growth factors on normal and tumour cell cycle, b) on the necessary condition for mitogenicity of lectins, c) on the stimulation of proliferation by mechanical deformation of cells.

The role of red blood cell deformability and Na,K-ATPase function in selected risk factors of cardiovascular diseases in humans: focus on hypertension, diabetes mellitus and hypercholesterolemia.

PubMed

Radosinska, J; Vrbjar, N

2016-09-19

Deformability of red blood cells (RBC) is the ability of RBC to change their shape in order to pass through narrow capillaries in circulation. Deterioration in deformability of RBC contributes to alterations in microcirculatory blood flow and delivery of oxygen to tissues. Several factors are responsible for maintenance of RBC deformability. One of them is the Na,K-ATPase known as crucial enzyme in maintenance of intracellular ionic homeostasis affecting thus regulation of cellular volume and consequently RBC deformability. Decreased deformability of RBC has been found to be the marker of adverse outcomes in cardiovascular diseases (CVD) and the presence of cardiovascular risk factors influences rheological properties of the blood. This review summarizes knowledge concerning the RBC deformability in connection with selected risk factors of CVD, including hypertension, hyperlipidemia, and diabetes mellitus, based exclusively on papers from human studies. We attempted to provide an update on important issues regarding the role of Na,K-ATPase in RBC deformability. In patients suffering from hypertension as well as diabetes mellitus the Na,K-ATPase appears to be responsible for the changes leading to alterations in RBC deformability. The triggering factor for changes of RBC deformability during hypercholesterolemia seems to be the increased content of cholesterol in erythrocyte membranes.

Impact of heat and water management on proton exchange membrane fuel cells degradation in automotive application

NASA Astrophysics Data System (ADS)

Nandjou, F.; Poirot-Crouvezier, J.-P.; Chandesris, M.; Blachot, J.-F.; Bonnaud, C.; Bultel, Y.

2016-09-01

In Proton Exchange Membrane Fuel Cells, local temperature is a driving force for many degradation mechanisms such as hygrothermal deformation and creep of the membrane, platinum dissolution and bipolar plates corrosion. In order to investigate and quantify those effects in automotive application, durability testing is conducted in this work. During the ageing tests, the local performance and temperature are investigated using in situ measurements of a printed circuit board. At the end of life, post-mortem analyses of the aged components are conducted. The experimental results are compared with the simulated temperature and humidity in the cell obtained from a pseudo-3D multiphysics model in order to correlate the observed degradations to the local conditions inside the stack. The primary cause of failure in automotive cycling is pinhole/crack formation in the membrane, induced by high variations of its water content over time. It is also observed that water condensation largely increases the probability of the bipolar plates corrosion while evaporation phenomena induce local deposits in the cell.

Profiling the erythrocyte membrane proteome isolated from patients diagnosed with chronic obstructive pulmonary disease.

PubMed

Alexandre, Bruno M; Charro, Nuno; Blonder, Josip; Lopes, Carlos; Azevedo, Pilar; Bugalho de Almeida, António; Chan, King C; Prieto, DaRue A; Issaq, Haleem; Veenstra, Timothy D; Penque, Deborah

2012-12-05

Structural and metabolic alterations in erythrocytes play an important role in the pathophysiology of Chronic Obstructive Pulmonary Disease (COPD). Whether these dysfunctions are related to the modulation of erythrocyte membrane proteins in patients diagnosed with COPD remains to be determined. Herein, a comparative proteomic profiling of the erythrocyte membrane fraction isolated from peripheral blood of smokers diagnosed with COPD and smokers with no COPD was performed using differential (16)O/(18)O stable isotope labeling. A total of 219 proteins were quantified as being significantly differentially expressed within the erythrocyte membrane proteomes of smokers with COPD and healthy smokers. Functional pathway analysis showed that the most enriched biofunctions were related to cell-to-cell signaling and interaction, hematological system development, immune response, oxidative stress and cytoskeleton. Chorein (VPS13A), a cytoskeleton related protein whose defects had been associated with the presence of cell membrane deformation of circulating erythrocytes was found to be down-regulated in the membrane fraction of erythrocytes obtained from COPD patients. Methemoglobin reductase (CYB5R3) was also found to be underexpressed in these cells, suggesting that COPD patients may be at higher risk for developing methemoglobinemia. This article is part of a Special Issue entitled: Integrated omics. Copyright © 2012 Elsevier B.V. All rights reserved.

Dynamics of Receptor-Mediated Nanoparticle Internalization into Endothelial Cells

PubMed Central

Gonzalez-Rodriguez, David; Barakat, Abdul I.

2015-01-01

Nanoparticles offer a promising medical tool for targeted drug delivery, for example to treat inflamed endothelial cells during the development of atherosclerosis. To inform the design of such therapeutic strategies, we develop a computational model of nanoparticle internalization into endothelial cells, where internalization is driven by receptor-ligand binding and limited by the deformation of the cell membrane and cytoplasm. We specifically consider the case of nanoparticles targeted against ICAM-1 receptors, of relevance for treating atherosclerosis. The model computes the kinetics of the internalization process, the dynamics of binding, and the distribution of stresses exerted between the nanoparticle and the cell membrane. The model predicts the existence of an optimal nanoparticle size for fastest internalization, consistent with experimental observations, as well as the role of bond characteristics, local cell mechanical properties, and external forces in the nanoparticle internalization process. PMID:25901833

Permeability of C2C12 myotube membranes is influenced by stretch velocity.

PubMed

Burkholder, Thomas J

2003-05-30

Mechanical signals are critical to the growth and maintenance of skeletal muscle, but the mechanism by which these signals are transduced by the cell remains unknown. This work examined the hypothesis that stretch conditions influence membrane permeability consistent with a role for membrane permeability in mechanotransduction. C2C12 myotubes were grown in conditions that encourage uniform alignment and subjected to uniform mechanical deformation in the presence of fluorescein labeled dextran to evaluate membrane permeability as a function of stretch amplitude and velocity. Within a physiologically relevant range of conditions, a complex interaction between the two aspects of stretch was observed, with velocity contributing most strongly at large stretch amplitudes. This suggests that membrane viscosity could contribute to mechanotransduction.

Deformation-induced release of ATP from erythrocytes in a poly(dimethylsiloxane)-based microchip with channels that mimic resistance vessels.

PubMed

Price, Alexander K; Fischer, David J; Martin, R Scott; Spence, Dana M

2004-08-15

The ability of nitric oxide to relax smooth muscle cells surrounding resistance vessels in vivo is well documented. Here, we describe a series of studies designed to quantify amounts of adenosine triphosphate (ATP), a known stimulus of NO production in endothelial cells, released from erythrocytes that are mechanically deformed as these cells traverse microbore channels in lithographically patterned microchips. Results indicate that micromolar amounts of ATP are released from erythrocytes flowing through channels having cross sectional dimensions of 60 x 38 micron (2.22 +/- 0.50 microM ATP). Microscopic images indicate that erythrocytes, when being pumped through the microchip channels, migrate toward the center of the channels, leaving a cell-free or skimming layer at the walls of the channel, a profile known to exist in circulatory vessels in vivo. A comparison of the amounts of ATP released from RBCs mechanically deformed in microbore tubing (2.54 +/- 0.15 microM) vs a microchip (2.59 +/- 0.32 microM) suggests that channels in microchips may serve as functional biomimics of the microvasculature. Control studies involving diamide, a membrane-stiffening agent, suggest that the RBC-derived ATP is not due to cell lysis but rather physical deformation.

A dielectric elastomer actuator coupled with water: snap-through instability and giant deformation

NASA Astrophysics Data System (ADS)

Godaba, Hareesh; Foo, Choon Chiang; Zhang, Zhi Qian; Khoo, Boo Cheong; Zhu, Jian

2015-04-01

A dielectric elastomer actuator is one class of soft actuators which can deform in response to voltage. Dielectric elastomer actuators coupled with liquid have recently been developed as soft pumps, soft lenses, Braille displays, etc. In this paper, we conduct experiments to investigate the performance of a dielectric elastomer actuator which is coupled with water. The membrane is subject to a constant water pressure, which is found to significantly affect the electromechanical behaviour of the membrane. When the pressure is small, the membrane suffers electrical breakdown before snap-through instability, and achieves a small voltage-induced deformation. When the pressure is higher to make the membrane near the verge of the instability, the membrane can achieve a giant voltage-induced deformation, with an area strain of 1165%. When the pressure is large, the membrane suffers pressure-induced snap-through instability and may collapse due to a large amount of liquid enclosed by the membrane. Theoretical analyses are conducted to interpret these experimental observations.

The role of purinergic signaling on deformation induced injury and repair responses of alveolar epithelial cells.

PubMed

Belete, Hewan A; Hubmayr, Rolf D; Wang, Shaohua; Singh, Raman-Deep

2011-01-01

Cell wounding is an important driver of the innate immune response of ventilator-injured lungs. We had previously shown that the majority of wounded alveolus resident cells repair and survive deformation induced insults. This is important insofar as wounded and repaired cells may contribute to injurious deformation responses commonly referred to as biotrauma. The central hypothesis of this communication states that extracellular adenosine-5' triphosphate (ATP) promotes the repair of wounded alveolus resident cells by a P2Y2-Receptor dependent mechanism. Using primary type 1 alveolar epithelial rat cell models subjected to micropuncture injury and/or deforming stress we show that 1) stretch causes a dose dependent increase in cell injury and ATP media concentrations; 2) enzymatic depletion of extracellular ATP reduces the probability of stretch induced wound repair; 3) enriching extracellular ATP concentrations facilitates wound repair; 4) purinergic effects on cell repair are mediated by ATP and not by one of its metabolites; and 5) ATP mediated cell salvage depends at least in part on P2Y2-R activation. While rescuing cells from wounding induced death may seem appealing, it is possible that survivors of membrane wounding become governors of a sustained pro-inflammatory state and thereby perpetuate and worsen organ function in the early stages of lung injury syndromes. Means to uncouple P2Y2-R mediated cytoprotection from P2Y2-R mediated inflammation and to test the preclinical efficacy of such an undertaking deserve to be explored.

Diagnostic tool for red blood cell membrane disorders: Assessment of a new generation ektacytometer.

PubMed

Da Costa, Lydie; Suner, Ludovic; Galimand, Julie; Bonnel, Amandine; Pascreau, Tiffany; Couque, Nathalie; Fenneteau, Odile; Mohandas, Narla

2016-01-01

Inherited red blood cell (RBC) membrane disorders, such as hereditary spherocytosis, elliptocytosis and hereditary ovalocytosis, result from mutations in genes encoding various RBC membrane and skeletal proteins. The RBC membrane, a composite structure composed of a lipid bilayer linked to a spectrin/actin-based membrane skeleton, confers upon the RBC unique features of deformability and mechanical stability. The disease severity is primarily dependent on the extent of membrane surface area loss. RBC membrane disorders can be readily diagnosed by various laboratory approaches that include RBC cytology, flow cytometry, ektacytometry, electrophoresis of RBC membrane proteins and genetics. The reference technique for diagnosis of RBC membrane disorders is the osmotic gradient ektacytometry. However, in spite of its recognition as the reference technique, this technique is rarely used as a routine diagnosis tool for RBC membrane disorders due to its limited availability. This may soon change as a new generation of ektacytometer has been recently engineered. In this review, we describe the workflow of the samples shipped to our Hematology laboratory for RBC membrane disorder analysis and the data obtained for a large cohort of French patients presenting with RBC membrane disorders using a newly available version of the ektacytomer. Copyright © 2015 Elsevier Inc. All rights reserved.

Linking actin networks and cell membrane via a reaction-diffusion-elastic description of nonlinear filopodia initiation.

PubMed

Ben Isaac, Eyal; Manor, Uri; Kachar, Bechara; Yochelis, Arik; Gov, Nir S

2013-08-01

Reaction-diffusion models have been used to describe pattern formation on the cellular scale, and traditionally do not include feedback between cellular shape changes and biochemical reactions. We introduce here a distinct reaction-diffusion-elasticity approach: The reaction-diffusion part describes bistability between two actin orientations, coupled to the elastic energy of the cell membrane deformations. This coupling supports spatially localized patterns, even when such solutions do not exist in the uncoupled self-inhibited reaction-diffusion system. We apply this concept to describe the nonlinear (threshold driven) initiation mechanism of actin-based cellular protrusions and provide support by several experimental observations.

Molecular dynamics simulations of membrane deformation induced by amphiphilic helices of Epsin, Sar1p, and Arf1

NASA Astrophysics Data System (ADS)

Li, Zhen-Lu

2018-03-01

The N-terminal amphiphilic helices of proteins Epsin, Sar1p, and Arf1 play a critical role in initiating membrane deformation. The interactions of these amphiphilic helices with the lipid membranes are investigated in this study by combining the all-atom and coarse-grained simulations. In the all-atom simulations, the amphiphilic helices of Epsin and Sar1p are found to have a shallower insertion depth into the membrane than the amphiphilic helix of Arf1, but remarkably, the amphiphilic helices of Epsin and Sar1p induce higher asymmetry in the lipid packing between the two monolayers of the membrane. The insertion depth of amphiphilic helix into the membrane is determined not only by the overall hydrophobicity but also by the specific distributions of polar and non-polar residues along the helix. To directly compare their ability to deform the membrane, the coarse-grained simulations are performed to investigate the membrane deformation under the insertion of multiple helices. Project supported by the National Natural Science Foundation of China (Grant Nos. 91427302 and 11474155).

Stretching of red blood cells by optical tweezers quantified by digital holographic microscopy

NASA Astrophysics Data System (ADS)

Cardenas, Nelson; Yu, Lingfeng; Mohanty, Samarendra K.

2011-03-01

Red blood cells (RBC) possess unique viscoelastic characteristics which allow them to pass through capillaries narrower than their size. Measurement of viscoelastic property of cells (e.g. RBC) in low-force regime is of high significance as it represents conditions of membrane fluctuation in response to physiological conditions. Estimation of visco-elastic properties of RBC requires measurement of extent of deformation in RBC subjected to known force. Optical tweezers, being gentle and absolutely sterile, are emerging as the tool of choice for application of localized force on cells. However, stretching of RBC in very low force regime has not been quantified. Further, though deformations in transverse directions have been measured, vertical deformations due to stretching of cells cannot be quantified by classical microscopic images. Here, we report realization of offaxis digital holographic microscopy (DHM) for highly sensitive axial changes in RBC shape due to stretching by optical tweezers without attaching microscopic beads. The RBC was stretched in axial direction with nanometer precision by change of divergence of the trapping beam. The obtained deformation patterns were compared with the axial position of the tweezers focus. Since the pathophysiology of progression of diseases like malaria and cancer is reflected in the biophysical (both mechanical and material) properties of the cells, it is possible to identify the changes by simultaneous measurement of refractive index and elasticity using this approach.

Fabrication and Characterization of Single Phase α-Alumina Membranes with Tunable Pore Diameters

PubMed Central

Masuda, Tatsuya; Asoh, Hidetaka; Haraguchi, Satoshi; Ono, Sachiko

2015-01-01

Nanoporous and single phase α-alumina membranes with pore diameters tunable over a wide range of approximately 60–350 nm were successfully fabricated by optimizing the conditions for anodizing, subsequent detachment, and heat treatment. The pore diameter increased and the cell diameter shrunk upon crystallization to α-alumina by approximately 20% and 3%, respectively, in accordance with the 23% volume shrinkage resulting from the change in density associated with the transformation from the amorphous state to α-alumina. Nevertheless, flat α-alumina membranes, each with a diameter of 25 mm and a thickness of 50 μm, were obtained without thermal deformation. The α-alumina membranes exhibited high chemical resistance in various concentrated acidic and alkaline solutions as well as when exposed to high temperature steam under pressure. The Young’s modulus and hardness of the single phase α-alumina membranes formed by heat treatment at 1250 °C were notably decreased compared to the corresponding amorphous membranes, presumably because of the nodular crystallite structure of the cell walls and the substantial increase in porosity. Furthermore, when used for filtration, the α-alumina membrane exhibited a level of flux higher than that of the commercial ceramic membrane. PMID:28788005

Investigating the Improved Aerodynamic Efficiency of Cambered Frames on Membrane MAV Wings

NASA Astrophysics Data System (ADS)

Wrist, Andrew; Zhang, Zheng; Hubner, Paul

2014-11-01

Previous research has demonstrated that membrane wings with cambered frames are more aerodynamically efficient than those with flat frames, despite passive dynamic membrane cambering for both. To help understand this aerodynamic benefit, this study compares the time-averaged membrane shape as well as membrane vibration frequency and amplitude for a group of wings with cambered frames. The frames were 3D printed with a hardened polymer material, and a silicon rubber membrane was attached to the top surface. The frame aspect ratio is two, comprised of two cells each with a cell aspect ratio of one. The rigid leading edge extended 20% of the chord, and the trailing edge was scalloped at 25%. Camber ranged from 2--6%, camber location from 40--60%, and airfoil thickness from 4--6%. Tests were performed in the University of Alabama's MAV wind tunnel at 10 m/s (Re = 50,000). High speed imaging results of the deformation and vibration will be discussed in context to airfoil and wing theory. National Science Foundation Grant Number: 1358991.

Structural properties of liposomes from digital holographic microscopy

NASA Astrophysics Data System (ADS)

Di Maio, Isabelle L.; Carl, Daniel; Langehanenberg, Patrik; Valenzuela, Stella M.; Battle, Andrew R.; Al Khazaaly, Sabah; Killingsworth, Murray; Kemper, Bjorn; von Bally, Gert; Martin, Donald K.

2006-01-01

We have constructed liposomes from L alpha Phosphatidylcholine (PC) lipids, which are biomimetic lipids similar to those present in the membranes of mammalian cells. We propose an advance in the use of liposomes, such as for drug delivery, to incorporate into the liposomal membranes transport proteins that have been extracted from the lipid membranes of mammalian cells. In this paper, we describe the usage of a novel optical microscope to characterize the nanomechanical properties of these liposomes. We have applied the technique of digital holographic microscopy, using an instrument recently developed at the University of Münster, Germany. This system enabled us to measure quantitatively the structural changes in liposomes. We have investigated the deformations of these biomimetic lipids comprising these liposomes by applying osmotic stresses, in order to gain insight into the membrane environment prior to incorporation of cloned membrane transport proteins. This control of the nanomechanical properties is important in the stresses transmitted to mechanosensitive ion channels that we have incorporated into the liposomal membranes. These liposomes provide transporting vesicles that respond to mechanical stresses, such as those that occur during implantation.

Dielectric elastomer membranes undergoing inhomogeneous deformation

NASA Astrophysics Data System (ADS)

He, Tianhu; Zhao, Xuanhe; Suo, Zhigang

2009-10-01

Dielectric elastomers are capable of large deformation subject to an electric voltage and are promising for use as actuators, sensors, and generators. Because of large deformation, nonlinear equations of states, and diverse modes of failure, modeling the process of electromechanical transduction has been challenging. This paper studies a membrane of a dielectric elastomer deformed into an out-of-plane axisymmetric shape, a configuration used in a family of commercial devices known as the universal muscle actuators. The kinematics of deformation and charging, together with thermodynamics, leads to equations that govern the state of equilibrium. Numerical results indicate that the field in the membrane can be very inhomogeneous, and that the membrane is susceptible to several modes of failure, including electrical breakdown, loss of tension, and rupture by stretch. Care is needed in the design to balance the requirements of averting various modes of failure while using the material efficiently.

Blood flow and blood cell interactions and migration in microvessels

NASA Astrophysics Data System (ADS)

Fedosov, Dmitry; Fornleitner, Julia; Gompper, Gerhard

2011-11-01

Blood flow in microcirculation plays a fundamental role in a wide range of physiological processes and pathologies in the organism. To understand and, if necessary, manipulate the course of these processes it is essential to investigate blood flow under realistic conditions including deformability of blood cells, their interactions, and behavior in the complex microvascular network which is characteristic for the microcirculation. We employ the Dissipative Particle Dynamics method to model blood as a suspension of deformable cells represented by a viscoelastic spring-network which incorporates appropriate mechanical and rheological cell-membrane properties. Blood flow is investigated in idealized geometries. In particular, migration of blood cells and their distribution in blood flow are studied with respect to various conditions such as hematocrit, flow rate, red blood cell aggregation. Physical mechanisms which govern cell migration in microcirculation and, in particular, margination of white blood cells towards the vessel wall, will be discussed. In addition, we characterize blood flow dynamics and quantify hemodynamic resistance. D.F. acknowledges the Humboldt Foundation for financial support.

Receptor-mediated membrane adhesion of lipid-polymer hybrid (LPH) nanoparticles studied by dissipative particle dynamics simulations

NASA Astrophysics Data System (ADS)

Li, Zhenlong; Gorfe, Alemayehu A.

2014-12-01

Lipid-polymer hybrid (LPH) nanoparticles represent a novel class of targeted drug delivery platforms that combine the advantages of liposomes and biodegradable polymeric nanoparticles. However, the molecular details of the interaction between LPHs and their target cell membranes remain poorly understood. We have investigated the receptor-mediated membrane adhesion process of a ligand-tethered LPH nanoparticle using extensive dissipative particle dynamics (DPD) simulations. We found that the spontaneous adhesion process follows a first-order kinetics characterized by two distinct stages: a rapid nanoparticle-membrane engagement, followed by a slow growth in the number of ligand-receptor pairs coupled with structural re-organization of both the nanoparticle and the membrane. The number of ligand-receptor pairs increases with the dynamic segregation of ligands and receptors toward the adhesion zone causing an out-of-plane deformation of the membrane. Moreover, the fluidity of the lipid shell allows for strong nanoparticle-membrane interactions to occur even when the ligand density is low. The LPH-membrane avidity is enhanced by the increased stability of each receptor-ligand pair due to the geometric confinement and the cooperative effect arising from multiple binding events. Thus, our results reveal the unique advantages of LPH nanoparticles as active cell-targeting nanocarriers and provide some general principles governing nanoparticle-cell interactions that may aid future design of LPHs with improved affinity and specificity for a given target of interest.

A novel function of WAVE in lamellipodia: WAVE1 is required for stabilization of lamellipodial protrusions during cell spreading.

PubMed

Yamazaki, Daisuke; Fujiwara, Takashi; Suetsugu, Shiro; Takenawa, Tadaomi

2005-05-01

When a cell spreads and moves, reorganization of the actin cytoskeleton pushes the cell membrane, and the resulting membrane protrusions create new points of contact with the substrate and generate the locomotive force. Membrane extension and adhesion to a substrate must be tightly coordinated for effective cell movement, but little is known about the mechanisms underlying these processes. WAVEs are critical regulators of Rac-induced actin reorganization. WAVE2 is essential for formation of lamellipodial structures at the cell periphery stimulated by growth factors, but it is thought that WAVE1 is dispensable for such processes in mouse embryonic fibroblasts (MEFs). Here we show a novel function of WAVE in lamellipodial protrusions during cell spreading. During spreading on fibronectin (FN), MEFs with knockouts (KOs) of WAVE1 and WAVE2 showed different membrane dynamics, suggesting that these molecules have distinct roles in lamellipodium formation. Formation of lamellipodial structures on FN was inhibited in WAVE2 KO MEFs. In contrast, WAVE1 is not essential for extension of lamellipodial protrusions but is required for stabilization of such structures. WAVE1-deficiency decreased the density of actin filaments and increased the speed of membrane extension, causing deformation of focal complex at the tip of spreading edges. Thus, at the tip of the lamellipodial protrusion, WAVE2 generates the membrane protrusive structures containing actin filaments, and modification by WAVE1 stabilizes these structures through cell-substrate adhesion. Coordination of WAVE1 and WAVE2 activities appears to be necessary for formation of proper actin structures in stable lamellipodia.

Nature-inspired multifunctional membrane fabricated by adaptive hybridization of PNIPAm and PPy

NASA Astrophysics Data System (ADS)

Kim, Hyejeong; Kim, Kiwoong; Lee, Sang Joon

2017-11-01

Specialized plant organs, such as guard cells of stomata, consist of soft materials with deformability and electrochemical properties in response to various environmental stimuli. Stimulus-responsive hydrogels with electrochemical properties are good candidates for imitating such functionalities having great potential in a wide range of applications. However, conductive hydrogels are usually mechanically rigid and the fabrication technology of structured hydrogels has low reproducibility. Here, inspired by stimulus-responsive functionalities of plants, a thermo-responsive multifunctional hybrid membrane (HM) is synthesized through the in situ hybridization of conductive poly(pyrrole)(PPy) on a photopolymerized poly(N-isopropylacrylamide)(PNIPAm) membrane. The various properties of the HM are investigated to characterize its multiple functions. In terms of morphology, the HM can be easily fabricated into various structures, and exhibits thermo-responsive deformability. In terms of functionality, it exhibits various electrical and charge responses to thermal stimuli. This simple and efficient fabrication method can be used as a promising platform for fabricating a variety of functional devices, such as actuators, biosensors, and filtration membranes. This work was supported by the National Research Foundation of Korea (NRF) Grant funded by the Korean government (MSIP) (No. 2017R1A2B3005415).

Growth of Walled Cells: From Shells to Vesicles

NASA Astrophysics Data System (ADS)

Boudaoud, Arezki

2003-07-01

The growth of isolated walled cells is investigated. Examples of such cells range from bacteria to giant algae, and include cochlear hair, plant root hair, fungi, and yeast cells. They are modeled as elastic shells containing a liquid. Cell growth is driven by fluid pressure and is is similar to a plastic deformation of the wall. The requirement of mechanical equilibrium leads to two new scaling laws for cell size that are in quantitative agreement with the compiled biological data. Given these results, possible shapes for growing cells are computed by analogy with those of vesicle membranes.

On the growth of walled cells: From shells to vesicles.

NASA Astrophysics Data System (ADS)

Boudaoud, Arezki

2003-03-01

The growth of isolated walled cells is investigated. Examples of such cells range from bacteria to giant algae, and include cochlear hair, plant root hair, fungi and yeast cells. They are modeled as elastic shells inflated by a liquid. Cell growth is driven by fluid pressure and is similar to a plastic deformation of the wall. The requirement of mechanical equilibrium leads to two new scaling laws for cell size that are in quantitative agreement with the compiled biological data. Given these results, possible shapes for growing cells are computed by analogy with those of vesicle membranes.

Radiographic contrast media alterate the localization of actin/band4.9 in the membrane cytoskeleton of human erythrocytes.

PubMed

Franke, R P; Scharnweber, T; Fuhrmann, R; Mrowietz, C; Wenzel, F; Krüger, A; Jung, F

2014-01-01

Different radiographic contrast media (RCM) were shown to induce morphological changes of blood cells (e.g. erythrocytes or thrombocytes) and endothelial cells. The echinocytic shape change of erythrocytes, particularly, affords alterations of the membrane cytoskeleton. The cytoskeleton plays a crucial role for the shape and deformability of the red blood cell. Disruption of the interaction between components of the red blood cell membrane cytoskeleton may cause a loss of structural and functional integrity of the membrane. In this study band4.9 and actin as components of the cytoskeletal junctional complex were examined in human erythrocytes after suspension in autologous plasma or in plasma RCM mixtures (30% v/v Iodixanol-320 or Iopromide-370) followed by a successive double staining with TRITC-/FITC-coupled monoclonal antibodies. After adding Iopromide-370 to the plasma in practically none of the cells the rounded conformation of the membrane cytoskeleton - as it appeared in cells suspended in autologous plasma - was found. In addition, Iopromide-370 induced thin lines and coarse knob-like structures of band4.9 at the cell periphery while most cell centers were devoid of band4.9, and a box-like arrangement of bands of band4.9. A dissociation between colours red (actin) and green (band4.9) occurred as well. In contrast, erythrocytes suspended in a plasma/Iodixanol-320 mixture showed a membrane cytoskeleton comparable to cells suspended in autologous plasma, Similar results were found with respect to the distribution of actin. This study revealed for the first time RCM-dependent differences in band4.9 activities as possible pathophysiological mechanism for the chemotoxicity of radiographic contrast media.

The Role of Purinergic Signaling on Deformation Induced Injury and Repair Responses of Alveolar Epithelial Cells

PubMed Central

Belete, Hewan A.; Hubmayr, Rolf D.; Wang, Shaohua; Singh, Raman-Deep

2011-01-01

Cell wounding is an important driver of the innate immune response of ventilator-injured lungs. We had previously shown that the majority of wounded alveolus resident cells repair and survive deformation induced insults. This is important insofar as wounded and repaired cells may contribute to injurious deformation responses commonly referred to as biotrauma. The central hypothesis of this communication states that extracellular adenosine-5′ triphosphate (ATP) promotes the repair of wounded alveolus resident cells by a P2Y2-Receptor dependent mechanism. Using primary type 1 alveolar epithelial rat cell models subjected to micropuncture injury and/or deforming stress we show that 1) stretch causes a dose dependent increase in cell injury and ATP media concentrations; 2) enzymatic depletion of extracellular ATP reduces the probability of stretch induced wound repair; 3) enriching extracellular ATP concentrations facilitates wound repair; 4) purinergic effects on cell repair are mediated by ATP and not by one of its metabolites; and 5) ATP mediated cell salvage depends at least in part on P2Y2-R activation. While rescuing cells from wounding induced death may seem appealing, it is possible that survivors of membrane wounding become governors of a sustained pro-inflammatory state and thereby perpetuate and worsen organ function in the early stages of lung injury syndromes. Means to uncouple P2Y2-R mediated cytoprotection from P2Y2-R mediated inflammation and to test the preclinical efficacy of such an undertaking deserve to be explored. PMID:22087324

Separation of red blood cells in deep deterministic lateral displacement devices

NASA Astrophysics Data System (ADS)

Kabacaoglu, Gokberk; Biros, George

2017-11-01

Microfluidic cell separation techniques are of great interest since they help rapid medical diagnoses and tests. Deterministic lateral displacement (DLD) is one of them. A DLD device consists of arrays of pillars. Main flow and alignment of the pillars define two different directions. Size-based separation of rigid spherical particles is possible as they follow one of these directions depending on their sizes. However, the separation of non-spherical deformable particles such as red blood cells (RBCs) is more complicated than that due to their intricate dynamics. We study the separation of RBCs in DLD using an in-house integral equation solver. We systematically investigate the effects of the interior fluid viscosity and the membrane elasticity of an RBC on its behavior. These mechanical properties of a cell determine its deformability, which can be altered by several diseases. We particularly consider deep devices in which an RBC can show rich dynamics such as tank-treading and tumbling. It turns out that strong hydrodynamic lift force moves the tank-treading cells along the pillars and downward force leads the tumbling ones to move with the flow. Thereby, deformability-based separation of RBCs is possible.

A Two-Dimensional Numerical Investigation of Transport of Malaria-Infected Red Blood Cells in Stenotic Microchannels

PubMed Central

Tao, Yong; Rongin, Uwitije; Xing, Zhongwen

2016-01-01

The malaria-infected red blood cells experience a significant decrease in cell deformability and increase in cell membrane adhesion. Blood hemodynamics in microvessels is significantly affected by the alteration of the mechanical property as well as the aggregation of parasitized red blood cells. In this study, we aim to numerically study the connection between cell-level mechanobiological properties of human red blood cells and related malaria disease state by investigating the transport of multiple red blood cell aggregates passing through microchannels with symmetric stenosis. Effects of stenosis magnitude, aggregation strength, and cell deformability on cell rheology and flow characteristics were studied by a two-dimensional model using the fictitious domain-immersed boundary method. The results indicated that the motion and dissociation of red blood cell aggregates were influenced by these factors and the flow resistance increases with the increase of aggregating strength and cell stiffness. Further, the roughness of the velocity profile was enhanced by cell aggregation, which considerably affected the blood flow characteristics. The study may assist us in understanding cellular-level mechanisms in disease development. PMID:28105411

On the role of ultra-thin oxide cathode synthesis on the functionality of micro-solid oxide fuel cells: Structure, stress engineering and in situ observation of fuel cell membranes during operation

NASA Astrophysics Data System (ADS)

Lai, Bo-Kuai; Kerman, Kian; Ramanathan, Shriram

Microstructure and stresses in dense La 0.6Sr 0.4Co 0.8Fe 0.2O 3 (LSCF) ultra-thin films have been investigated to increase the physical thickness of crack-free cathodes and active area of thermo-mechanically robust micro-solid oxide fuel cell (μSOFC) membranes. Processing protocols employ low deposition rates to create a highly granular nanocrystalline microstructure in LSCF thin films and high substrate temperatures to produce linear temperature-dependent stress evolution that is dominated by compressive stresses in μSOFC membranes. Insight and trade-off on the synthesis are revealed by probing microstructure evolution and electrical conductivity in LSCF thin films, in addition to in situ monitoring of membrane deformation while measuring μSOFC performance at varying temperatures. From these studies, we were able to successfully fabricate failure-resistant square μSOFC (LSCF/YSZ/Pt) membranes with width of 250 μm and crack-free cathodes with thickness of ∼70 nm. Peak power density of ∼120 mW cm -2 and open circuit voltage of ∼0.6 V at 560 °C were achieved on a μSOFC array chip containing ten such membranes. Mechanisms affecting fuel cell performance are discussed. Our results provide fundamental insight to pathways of microstructure and stress engineering of ultra-thin, dense oxide cathodes and μSOFC membranes.

Opto-mechanical analysis of nonlinear elastomer membrane deformation under hydraulic pressure for variable-focus liquid-filled microlenses.

PubMed

Choi, Seung Tae; Son, Byeong Soo; Seo, Gye Won; Park, Si-Young; Lee, Kyung-Sick

2014-03-10

Nonlinear large deformation of a transparent elastomer membrane under hydraulic pressure was analyzed to investigate its optical performance for a variable-focus liquid-filled membrane microlens. In most membrane microlenses, actuators control the hydraulic pressure of optical fluid so that the elastomer membrane together with the internal optical fluid changes its shape, which alters the light path of the microlens to adapt its optical power. A fluid-structure interaction simulation was performed to estimate the transient behavior of the microlens under the operation of electroactive polymer actuators, demonstrating that the viscosity of the optical fluid successfully stabilizes the fluctuations within a fairly short period of time during dynamic operations. Axisymmetric nonlinear plate theory was used to calculate the deformation profile of the membrane under hydrostatic pressure, with which optical characteristics of the membrane microlens were estimated. The effects of gravitation and viscoelastic behavior of the elastomer membrane on the optical performance of the membrane microlens were also evaluated with finite element analysis.

Effects of membrane deformability and bond formation/dissociation rates on adhesion dynamics of a spherical capsule in shear flow.

PubMed

Zhang, Ziying; Du, Jun; Wei, Zhengying; Wang, Zhen; Li, Minghui

2018-02-01

Cellular adhesion plays a critical role in biological systems and biomedical applications. Cell deformation and biophysical properties of adhesion molecules are of significance for the adhesion behavior. In the present work, dynamic adhesion of a deformable capsule to a planar substrate, in a linear shear flow, is numerically simulated to investigate the combined influence of membrane deformability (quantified by the capillary number) and bond formation/dissociation rates on the adhesion behavior. The computational model is based on the immersed boundary-lattice Boltzmann method for the capsule-fluid interaction and a probabilistic adhesion model for the capsule-substrate interaction. Three distinct adhesion states, detachment, rolling adhesion and firm adhesion, are identified and presented in a state diagram as a function of capillary number and bond dissociation rate. The impact of bond formation rate on the state diagram is further investigated. Results show that the critical bond dissociation rate for the transition of rolling or firm adhesion to detachment is strongly related to the capsule deformability. At the rolling-adhesion state, smaller off rates are needed for larger capillary number to increase the rolling velocity and detach the capsule. In contrast, the critical off rate for firm-to-detach transition slightly increases with the capillary number. With smaller on rate, the effect of capsule deformability on the critical off rates is more pronounced and capsules with moderate deformability are prone to detach by the shear flow. Further increasing of on rate leads to large expansion of both rolling-adhesion and firm-adhesion regions. Even capsules with relatively large deformability can maintain stable rolling adhesion at certain off rate.

PMCA activity and membrane tubulin affect deformability of erythrocytes from normal and hypertensive human subjects.

PubMed

Monesterolo, Noelia E; Nigra, Ayelen D; Campetelli, Alexis N; Santander, Verónica S; Rivelli, Juan F; Arce, Carlos A; Casale, Cesar H

2015-11-01

Our previous studies demonstrated formation of a complex between acetylated tubulin and brain plasma membrane Ca(2+)-ATPase (PMCA), and the effect of the lipid environment on structure of this complex and on PMCA activity. Deformability of erythrocytes from hypertensive human subjects was reduced by an increase in membrane tubulin content. In the present study, we examined the regulation of PMCA activity by tubulin in normotensive and hypertensive erythrocytes, and the effect of exogenously added diacylglycerol (DAG) and phosphatidic acid (PA) on erythrocyte deformability. Some of the key findings were that: (i) PMCA was associated with tubulin in normotensive and hypertensive erythrocytes, (ii) PMCA enzyme activity was directly correlated with erythrocyte deformability, and (iii) when tubulin was present in the erythrocyte membrane, treatment with DAG or PA led to increased deformability and associated PMCA activity. Taken together, our findings indicate that PMCA activity is involved in deformability of both normotensive and hypertensive erythrocytes. This rheological property of erythrocytes is affected by acetylated tubulin and its lipid environment because both regulate PMCA activity. Copyright © 2015 Elsevier B.V. All rights reserved.

Modeling of interactions between nanoparticles and cell membranes

NASA Astrophysics Data System (ADS)

Ban, Young-Min

Rapid development of nanotechnology and ability to manufacture materials and devices with nanometer feature size leads to exciting innovations in many areas including the medical and electronic fields. However, the possible health and environmental impacts of manufactured nanomaterials are not fully known. Recent experimental reports suggest that some of the manufactured nanomaterials, such as fullerenes and carbon nanotubes, are highly toxic even in small concentrations. The goal of the current work is to understand the mechanisms responsible for the toxicity of nanomaterials. In the current study coarse-grained molecular dynamics simulations are employed to investigate the interactions between NPs and cellular membranes at a molecular level. One of the possible toxicity mechanisms of the nanomaterials is membrane disruption. Possibility of membrane disruption exposed to the manufactured nanomaterials are examined by considering chemical reactions and non-reactive physical interactions as chemical as well as physical mechanisms. Mechanisms of transport of carbon-based nanoparticles (fullerene and its derivative) across a phospholipid bilayer are investigated. The free energy profile is obtained using constrained simulations. It is shown that the considered nanoparticles are hydrophobic and therefore they tend to reside in the interior of the lipid bilayer. In addition, the dynamics of the membrane fluctuations is significantly affected by the nanoparticles at the bilayer-water interface. The hydrophobic interaction between the particles and membrane core induces the strong coupling between the nanoparticle motion and membrane deformation. It is observed that the considered nanoparticles affect several physical properties of the membrane. The nanoparticles embedded into the membrane interior lead to the membrane softening, which becomes more significant with increase in CNT length and concentration. The lateral pressure profile and membrane energy in the membrane containing the nanoparticles exhibit localized perturbation around the nanoparticle. The nanoparticles are not likely to affect membrane protein function by the weak perturbation of the internal stress in the membrane. Due to the short-ranged interactions between the nanoparticles, the nanoparticles would not form aggregates inside membranes. The effect of lipid peroxidation on cell membrane deformation is assessed. The peroxidized lipids introduce a perturbation to the internal structure of the membrane leading to higher amplitude of the membrane fluctuations. Higher concentration of the peroxidized lipids induces more significant perturbation. Cumulative effects of lipid peroxidation caused by nanoparticles are examined for the first time. The considered amphiphilic particle appears to reduce the perturbation of the membrane structure at its equilibrium position inside the peroxidized membrane. This suggests a possibility of antioxidant effect of the nanoparticle.

Direct visualization of the arterial wall water permeability barrier using CARS microscopy

PubMed Central

Lucotte, Bertrand M.; Powell, Chloe; Knutson, Jay R.; Combs, Christian A.; Malide, Daniela; Yu, Zu-Xi; Knepper, Mark; Patel, Keval D.; Pielach, Anna; Johnson, Errin; Borysova, Lyudmyla; Balaban, Robert S.

2017-01-01

The artery wall is equipped with a water permeation barrier that allows blood to flow at high pressure without significant water leak. The precise location of this barrier is unknown despite its importance in vascular function and its contribution to many vascular complications when it is compromised. Herein we map the water permeability in intact arteries, using coherent anti-Stokes Raman scattering (CARS) microscopy and isotopic perfusion experiments. Generation of the CARS signal is optimized for water imaging with broadband excitation. We identify the water permeation barrier as the endothelial basolateral membrane and show that the apical membrane is highly permeable. This is confirmed by the distribution of the AQP1 water channel within endothelial membranes. These results indicate that arterial pressure equilibrates within the endothelium and is transmitted to the supporting basement membrane and internal elastic lamina macromolecules with minimal deformation of the sensitive endothelial cell. Disruption of this pressure transmission could contribute to endothelial cell dysfunction in various pathologies. PMID:28373558

Evaluation of surface deformability of lipid nanocapsules by drop tensiometer technique, and its experimental assessment by dialysis and tangential flow filtration.

PubMed

Hirsjärvi, Samuli; Bastiat, Guillaume; Saulnier, Patrick; Benoît, Jean-Pierre

2012-09-15

Deformability of nanoparticles might affect their behaviour at biological interfaces. Lipid nanocapsules (LNCs) are semi-solid particles resembling a hybrid of polymer nanoparticles and liposomes. Deformability of LNCs of different sizes was modelled by drop tensiometer technique. Two purification methods, dialysis and tangential flow filtration (TFF), were applied to study experimental behaviour and deformability of LNCs in order to evaluate if these properties contributed to membrane passing. Rheological parameters obtained from the drop tensiometer analysis suggested decreasing surface deformability of LNCs with increase in diameter. Dialysis results showed that up to 10% of LNCs can be lost during the process (e.g. membrane accumulation) but no clear evidence of the membrane passing was observed. Instead, LNCs with initial size and size distribution could be found in the TFF filtrate although molecular weight cut-off (MWCO) of the membrane used was smaller than the LNC diameter. Copyright © 2012 Elsevier B.V. All rights reserved.

Characterizations of individual human red blood cells from patients with diabetes mellitus (Conference Presentation)

NASA Astrophysics Data System (ADS)

Lee, SangYun; Jang, Seongsoo; Park, HyunJoo; Park, YongKeun

2016-03-01

We systematically measure the morphological, biochemical, and biomechanical properties of individual human red blood cells (RBCs) from patients with diabetes mellitus using quantitative phase imaging technique to characterize the diabetic red cells with respect to those of the healthy. The 3-D refractive index tomograms and 2-D dynamic membrane fluctuation maps of individual RBCs are reconstructed from a set of the retrieved complex optical fields at various laser incidence angles using the Common-path diffraction optical tomography, from which volume, surface area, sphericity, hemoglobin (Hb) concentration, Hb content, and membrane fluctuation are obtained simultaneously. The correlative relations among the retrieved red cell indices of diabetic and healthy RBCs are also investigated with capabilities of individual cell measurement. As expected, there are no significant alterations in morphologies (cellular volumes, surface area, and sphericity) between diabetic and healthy RBCs. However, despite the minute mean corpuscular Hb differences in cell blood count datasheet, the measured Hb concentrations and Hb contents of diabetic RBCs are statistically higher than those of healthy RBCs, which might be related to the glycation of Hb molecules by hyperglycemia. Meanwhile, the membrane fluctuations of diabetic RBCs are clearly diminished compared to healthy red cells, implying the significantly decreased RBC deformability. In particular, it seems that the membrane fluctuations have mild negative relationships with the reported HbA1c levels.

Mechanical algal disruption for efficient biodiesel extraction

NASA Astrophysics Data System (ADS)

Krehbiel, Joel David

Biodiesel from algae provides several benefits over current biodiesel feedstocks, but the energy requirements of processing algae into a useable fuel are currently so high as to be prohibitive. One route to improving this is via disruption of the cells prior to lipid extraction, which can significantly increase energy recovery. Unfortunately, several obvious disruption techniques require more energy than can be gained. This dissertation examines the use of microbubbles to improve mechanical disruption of algal cells using experimental, theoretical, and computational methods. New laboratory experiments show that effective ultrasonic disruption of algae is achieved by adding microbubbles to an algal solution. The configuration studied flows the solution through a tube and insonifies a small section with a high-pressure ultrasound wave. Previous biomedical research has shown effective cell membrane damage on animal cells with similar methods, but the present research is the first to extend such study to algal cells. Results indicate that disruption increases with peak negative pressure between 1.90 and 3.07 MPa and with microbubble concentration up to 12.5 x 107 bubbles/ml. Energy estimates of this process suggest that it requires only one-fourth the currently most-efficient laboratory-scale disruption process. Estimates of the radius near each bubble that causes disruption (i.e. the disruption radius) suggest that it increases with peak negative pressure and is near 9--20 microm for all cases tested. It is anticipated that these procedures can be designed for better efficiency and efficacy, which will be facilitated by identifying the root mechanisms of the bubble-induced disruption. We therefore examine whether bubble expansion alone creates sufficient cell deformation for cell rupture. The spherically-symmetric Marmottant model for bubble dynamics allows estimation of the flow regime under experimental conditions. Bubble expansion is modeled as a point source of mass at the bubble center, and if the bubble-to-cell spacing is much larger than the cell radius, the flow around the cell is approximately extensional in the cell's frame of reference. It is known that the present algae are quasi-spherical with cytoplasmic viscosity approximately 100 times that of water, so the cell is approximated as a viscous sphere. Thus, conditions that cause cell disruption from an expanding microbubble are modeled as either steady inviscid extensional flow or steady point source flow over a viscous sphere. In the inviscid extensional flow model, the flow inside the sphere is dominated by viscous forces so the Stokes equation is solved with matched stresses at the sphere surface from the exterior inviscid extensional flow. The short-time deformation of the sphere surface suggests that inviscid extensional flow is insufficient to disrupt cells. This indicates that asymmetry of the flow over the sphere may be required to provide sufficient surface areal strain to rupture the cell. In a more detailed model, the bubble expansion is modeled as an expansion near a viscous sphere using finite element software. For conditions similar to those seen in the experiment, deformation shows similar scaling to disruption. The deformation in this model is significantly higher than predicted from the inviscid extensional flow model due to the effect of asymmetric flow on the cell membrane. Estimates suggest 21% average areal strain on the algal membrane is required to disrupt algal cells, and this result agrees well with areal strains typically required to disrupt cell membranes although the actual value would be lessened by the effect of an elastic membrane, which is neglected in the present model. The local areal strain on the sphere surface is a maximum closest to the point source, and there is compressive strain near theta = +/-pi/4 with theta the angle from the line between the cell center and the point source. The maximum local areal strain shows less sensitivity to the viscosity of the interior fluid than the average areal strain. Overall, the dissertation lays the groundwork for more efficient algal disruption through the judicious use of microbubbles. Separation of bubble generation and bubble growth provides the ability to improve the efficiency of each process and localize energy. Results suggest that effective disruption can occur by pulsing high-pressure ultrasound waves to a solution of cells co-suspended with microbubbles. The models are thought to represent basic phenomenological mechanisms of disruption that could be exploited to improve the overall energy efficiency of schemes. Analysis suggests that extensional flow alone cannot be the cause of cell disruption near an expanding microbubble. Additionally, this work provides an estimate of the areal strain required disrupt an algal cell membrane. This research suggests a couple routes toward reducing the energy required for production of algal biodiesel.

Controlled vesicle deformation and lysis by single oscillating bubbles

NASA Astrophysics Data System (ADS)

Marmottant, Philippe; Hilgenfeldt, Sascha

2003-05-01

The ability of collapsing (cavitating) bubbles to focus and concentrate energy, forces and stresses is at the root of phenomena such as cavitation damage, sonochemistry or sonoluminescence. In a biomedical context, ultrasound-driven microbubbles have been used to enhance contrast in ultrasonic images. The observation of bubble-enhanced sonoporation-acoustically induced rupture of membranes-has also opened up intriguing possibilities for the therapeutic application of sonoporation as an alternative to cell-wall permeation techniques such as electroporation and particle guns. However, these pioneering experiments have not been able to pinpoint the mechanism by which the violently collapsing bubble opens pores or larger holes in membranes. Here we present an experiment in which gentle (linear) bubble oscillations are sufficient to achieve rupture of lipid membranes. In this regime, the bubble dynamics and the ensuing sonoporation can be accurately controlled. The use of microbubbles as focusing agents makes acoustics on the micrometre scale (microacoustics) a viable tool, with possible applications in cell manipulation and cell-wall permeation as well as in microfluidic devices.

Dynamic motion of red blood cells in simple shear flow

NASA Astrophysics Data System (ADS)

Sui, Y.; Chew, Y. T.; Roy, P.; Cheng, Y. P.; Low, H. T.

2008-11-01

A three-dimensional numerical model is proposed to simulate the dynamic motion of red blood cells (RBCs) in simple shear flow. The RBCs are approximated by ghost cells consisting of Newtonian liquid drops enclosed by Skalak membranes which take into account the membrane shear elasticity and the membrane area incompressibility. The RBCs have an initially biconcave discoid resting shape, and the internal liquid is assumed to have the same physical properties as the matrix fluid. The simulation is based on a hybrid method, in which the immersed boundary concept is introduced into the framework of the lattice Boltzmann method, and a finite element model is incorporated to obtain the forces acting on the nodes of the cell membrane which is discretized into flat triangular elements. The dynamic motion of RBCs is investigated in simple shear flow under a broad range of shear rates. At large shear rates, the cells are found to carry out a swinging motion, in which periodic inclination oscillation and shape deformation superimpose on the membrane tank treading motion. With the shear rate decreasing, the swinging amplitude of the cell increases, and finally triggers a transition to tumbling motion. This is the first direct numerical simulation that predicts both the swinging motion of the RBCs and the shear rate induced transition, which have been observed in a recent experiment. It is also found that as the mode changes from swinging to tumbling, the apparent viscosity of the suspension increases monotonically.

Probing the Bioeffects of Cavitation at the Single-Cell Level

NASA Astrophysics Data System (ADS)

Yuan, Fang

The primary goal of this dissertation research is to develop an experimental system and associated techniques that can be used to investigate the bioeffects produced by cavitation bubbles at the single cell level. Such information has been lacking due to the randomness and complexity in cavitation inception and subsequent bubble-bubble interaction generated by an acoustic field typically used in therapeutic ultrasound applications. Connection between cavitation activities and bioeffects produced in cells nearby presents another challenge that has not been resolved satisfactorily. In this work, we developed a laser-based system for generating tandem bubbles with a maximum diameter about 50 microm (i.e., on the scale of a single cell) in a microfluidic channel of 25 microm in height and 800 microm in width. We further developed techniques for micropatterning of individual gold dots (15 nm thick and 6 microm in diameter) used for bubble generation, which are precisely aligned at various stand-off distances (SD) from individual islands (32 x 32 microm2) coated with fibronectin used for cell adhesion. The dynamics of tandem bubble interaction with resultant jet formation, microstreaming and vortex flow in the microfluidic channel were captured by high-speed imaging and particle image velocimetry (PIV). The deformation of the target cell was recorded by high-speed imaging as well (using a second camera) immediately after the tandem bubble interaction and assessment of membrane strain was aided with 2 microm sized polystyrene beads attached to the cell membrane. Membrane poration was characterized by uptake of fluorescent propodium iodide (PI) into the target cell, from which the normalized maximum pore size was estimated. Using this experimental system, we have observed the complete process of bubble-bubble interaction with resultant jetting flow, cell deformation, and localized pinpoint membrane rupture with progressive diffusion of macromolecules into the target cell. Furthermore, we observed a clear SD dependence in the treatment outcome produced by the tandem bubbles. At short SD of 10 microm, all treated cells underwent necrosis with high yet unsaturated level of PI uptake, indicating that the cell could not reseal the poration site. At intermediate SD of 20 ˜ 30 microm, 58% to 80% of the cells were observed to have repairable membrane poration with low to medium but saturated level of PI uptake. At long SD of 40 microm, no detectable PI uptake was observed, corresponding to no membrane compromise. Within the repairable membrane poration group, the sub-population of cells that eventually survived without apoptosis increased from about 9% at SD of 20 microm with strong adhesion to about 70% at SD of 30 microm with no adhesion at the leading edge facing the jetting flow. The maximum PI uptake, pore size, and membrane strain estimated could vary by more than an order of magnitude, which is similar to the magnitude of variations in pore size (0.2 ˜ 2 microm) produced by tandem bubbles observed by SEM. The large principal strain (> 500%) with associated high strain-rate (> 106·s -1) produced by the tandem bubbles provide a unique tool to examine the bioeffects of cavitation at the single cell level and potentially a diverse range of applications to be explored.

Statolith sedimentation kinetics and force transduction to the cortical endoplasmic reticulum in gravity-sensing Arabidopsis columella cells.

PubMed

Leitz, Guenther; Kang, Byung-Ho; Schoenwaelder, Monica E A; Staehelin, L Andrew

2009-03-01

The starch statolith hypothesis of gravity sensing in plants postulates that the sedimentation of statoliths in specialized statocytes (columella cells) provides the means for converting the gravitational potential energy into a biochemical signal. We have analyzed the sedimentation kinetics of statoliths in the central S2 columella cells of Arabidopsis thaliana. The statoliths can form compact aggregates with gap sizes between statoliths approaching <30 nm. Significant intra-aggregate sliding motions of individual statoliths suggest a contribution of hydrodynamic forces to the motion of statoliths. The reorientation of the columella cells accelerates the statoliths toward the central cytoplasm within <1 s of reorientation. During the subsequent sedimentation phase, the statoliths tend to move at a distance to the cortical endoplasmic reticulum (ER) boundary and interact only transiently with the ER. Statoliths moved by laser tweezers against the ER boundary experience an elastic lift force upon release from the optical trap. High-resolution electron tomography analysis of statolith-to-ER contact sites indicate that the weight of statoliths is sufficient to locally deform the ER membranes that can potentially activate mechanosensitive ion channels. We suggest that in root columella cells, the transduction of the kinetic energy of sedimenting statoliths into a biochemical signal involves a combination of statolith-driven motion of the cytosol, statolith-induced deformation of the ER membranes, and a rapid release of kinetic energy from the ER during reorientation to activate mechanosensitive sites within the central columella cells.

Statolith Sedimentation Kinetics and Force Transduction to the Cortical Endoplasmic Reticulum in Gravity-Sensing Arabidopsis Columella Cells[W][OA

PubMed Central

Leitz, Guenther; Kang, Byung-Ho; Schoenwaelder, Monica E.A.; Staehelin, L. Andrew

2009-01-01

The starch statolith hypothesis of gravity sensing in plants postulates that the sedimentation of statoliths in specialized statocytes (columella cells) provides the means for converting the gravitational potential energy into a biochemical signal. We have analyzed the sedimentation kinetics of statoliths in the central S2 columella cells of Arabidopsis thaliana. The statoliths can form compact aggregates with gap sizes between statoliths approaching <30 nm. Significant intra-aggregate sliding motions of individual statoliths suggest a contribution of hydrodynamic forces to the motion of statoliths. The reorientation of the columella cells accelerates the statoliths toward the central cytoplasm within <1 s of reorientation. During the subsequent sedimentation phase, the statoliths tend to move at a distance to the cortical endoplasmic reticulum (ER) boundary and interact only transiently with the ER. Statoliths moved by laser tweezers against the ER boundary experience an elastic lift force upon release from the optical trap. High-resolution electron tomography analysis of statolith-to-ER contact sites indicate that the weight of statoliths is sufficient to locally deform the ER membranes that can potentially activate mechanosensitive ion channels. We suggest that in root columella cells, the transduction of the kinetic energy of sedimenting statoliths into a biochemical signal involves a combination of statolith-driven motion of the cytosol, statolith-induced deformation of the ER membranes, and a rapid release of kinetic energy from the ER during reorientation to activate mechanosensitive sites within the central columella cells. PMID:19276442

Deformation-Induced Precession of a Robot Moving on Curved Space

NASA Astrophysics Data System (ADS)

Li, Shengkai; Aydin, Yasemin; Lofaro, Olivia; Rieser, Jennifer; Goldman, Daniel

Previous studies have demonstrated that passive particles rolling on a deformed surface can mimic aspects of general relativity [Ford et al, AJP, 2015]. However, these systems are dissipative. To explore steady-state dynamics, we study the movement of a self-propelled robot car on a large deformable elastic membrane: a spandex sheet stretched over a metal frame with a diameter of 2.5 m. Two wheels in the rear of the car are differentially-driven by a DC motor, and a caster in the front helps maintain directional stability; in the absence of curvature the car drives straight. A linear actuator attached below the membrane allows for controlled deformation at the center of the membrane. We find that closed elliptic orbits occur when the membrane is highly depressed ( 10 cm). However, when the center is only slightly indented, the elliptical orbits precess at a rate depending on the orbit shape and the depression. Remarkably, this dynamic is well described by the Schwarzschild metric solution, typically used to describe the effects of gravity on bodies orbiting a massive object. Experiments with multiple cars reveal complex interactions that are mediated through car-induced deformations of the membrane.

Effects of deformability of RBCs on their dynamics and blood flow passing through a stenosed microvessel: an immersed boundary-lattice Boltzmann approach

NASA Astrophysics Data System (ADS)

Alizadeh, As'ad; Dadvand, Abdolrahman

2018-02-01

In this paper, the motion of high deformable (healthy) and low deformable (sick) red blood cells in a microvessel with and without stenosis is simulated using a combined lattice Boltzmann-immersed boundary method. The RBC is considered as neo-Hookean elastic membrane with bending resistance. The motion and deformation of the RBC under different values of the Reynolds number are evaluated. In addition, the variations of blood flow resistance and time-averaged pressure due to the motion and deformation of the RBC are assessed. It was found that a healthy RBC moves faster than a sick one. The apparent viscosity and blood flow resistance are greater for the case involving the sick RBC. Blood pressure at the presence of stenosis and low deformable RBC increases, which is thought of as the reason of many serious diseases including cardiovascular diseases. As the Re number increases, the RBC deforms further and moves easier and faster through the stenosis. The results of this study were compared to the available experimental and numerical results, and good agreements were observed.

The Effect of Sepsis on the Erythrocyte.

PubMed

Bateman, Ryon M; Sharpe, Michael D; Singer, Mervyn; Ellis, Christopher G

2017-09-08

Sepsis induces a wide range of effects on the red blood cell (RBC). Some of the effects including altered metabolism and decreased 2,3-bisphosphoglycerate are preventable with appropriate treatment, whereas others, including decreased erythrocyte deformability and redistribution of membrane phospholipids, appear to be permanent, and factors in RBC clearance. Here, we review the effects of sepsis on the erythrocyte, including changes in RBC volume, metabolism and hemoglobin's affinity for oxygen, morphology, RBC deformability (an early indicator of sepsis), antioxidant status, intracellular Ca 2+ homeostasis, membrane proteins, membrane phospholipid redistribution, clearance and RBC O₂-dependent adenosine triphosphate efflux (an RBC hypoxia signaling mechanism involved in microvascular autoregulation). We also consider the causes of these effects by host mediated oxidant stress and bacterial virulence factors. Additionally, we consider the altered erythrocyte microenvironment due to sepsis induced microvascular dysregulation and speculate on the possible effects of RBC autoxidation. In future, a better understanding of the mechanisms involved in sepsis induced erythrocyte pathophysiology and clearance may guide improved sepsis treatments. Evidence that small molecule antioxidants protect the erythrocyte from loss of deformability, and more importantly improve septic patient outcome suggest further research in this area is warranted. While not generally considered a critical factor in sepsis, erythrocytes (and especially a smaller subpopulation) appear to be highly susceptible to sepsis induced injury, provide an early warning signal of sepsis and are a factor in the microvascular dysfunction that has been associated with organ dysfunction.

Biotoxicity of TiO2 Nanoparticles on Raphidocelis subcapitata Microalgae Exemplified by Membrane Deformation

PubMed Central

Ozkaleli, Merve; Erdem, Ayca

2018-01-01

TiO2 nanoparticles (NPs), which are mainly used in consumer products (mostly cosmetics), have been found to cause ecotoxic effects in the aquatic environment. The green algae Raphidocelis subcapitata, as a representative of primary producers of the freshwater ecosystem, has been frequently used to study the effects of metal oxide NPs. An ecotoxicity study was conducted herein to investigate the effects of TiO2 NPs on survival and membrane deformation of algal cells. Five different concentrations of nano-TiO2 particles (1, 10, 50, 100 and 500 mg/L) were prepared in synthetic surface water samples with five different water quality characteristics (pH 6.4–8.4, hardness 10–320 mg CaCO3/L, ionic strength 0.2–8 mM, and alkalinity 10–245 mg CaCO3/L). Results showed a significant increase in the hydrodynamic diameter of NPs with respect to both NP concentrations and ionic content of the test system. A soft synthetic freshwater system at pH 7.3 ± 0.2 appeared to provide the most effective water type, with more than 95% algal mortality observed at 50, 100 and 500 mg/L NP concentrations. At high exposure concentrations, increased malondialdehyde formations were observed. Moreover, due to membrane deformation, TEM images correlated the uptake of the NPs. PMID:29495534

Asymmetrical Polyhedral Configuration of Giant Vesicles Induced by Orderly Array of Encapsulated Colloidal Particles

PubMed Central

Natsume, Yuno; Toyota, Taro

2016-01-01

Giant vesicles (GVs) encapsulating colloidal particles by a specific volume fraction show a characteristic configuration under a hypertonic condition. Several flat faces were formed in GV membrane with orderly array of inner particles. GV shape changed from the spherical to the asymmetrical polyhedral configuration. This shape deformation was derived by entropic interaction between inner particles and GV membrane. Because a part of inner particles became to form an ordered phase in the region neighboring the GV membrane, free volume for the other part of particles increased. Giant vesicles encapsulating colloidal particles were useful for the model of “crowding effect” which is the entropic interaction in the cell. PMID:26752650

Asymmetrical Polyhedral Configuration of Giant Vesicles Induced by Orderly Array of Encapsulated Colloidal Particles.

PubMed

Natsume, Yuno; Toyota, Taro

2016-01-01

Giant vesicles (GVs) encapsulating colloidal particles by a specific volume fraction show a characteristic configuration under a hypertonic condition. Several flat faces were formed in GV membrane with orderly array of inner particles. GV shape changed from the spherical to the asymmetrical polyhedral configuration. This shape deformation was derived by entropic interaction between inner particles and GV membrane. Because a part of inner particles became to form an ordered phase in the region neighboring the GV membrane, free volume for the other part of particles increased. Giant vesicles encapsulating colloidal particles were useful for the model of "crowding effect" which is the entropic interaction in the cell.

Hydrodynamic mobility of confined polymeric particles, vesicles, and cancer cells in a square microchannel.

PubMed

Ahmmed, Shamim M; Suteria, Naureen S; Garbin, Valeria; Vanapalli, Siva A

2018-01-01

The transport of deformable objects, including polymer particles, vesicles, and cells, has been a subject of interest for several decades where the majority of experimental and theoretical studies have been focused on circular tubes. Due to advances in microfluidics, there is a need to study the transport of individual deformable particles in rectangular microchannels where corner flows can be important. In this study, we report measurements of hydrodynamic mobility of confined polymeric particles, vesicles, and cancer cells in a linear microchannel with a square cross-section. Our operating conditions are such that the mobility is measured as a function of geometric confinement over the range 0.3 <  λ  < 1.5 and at specified particle Reynolds numbers that are within 0.1 < Re p  < 2.5. The experimental mobility data of each of these systems is compared with the circular-tube theory of Hestroni, Haber, and Wacholder [J. Fluid Mech. 41 , 689-705 (1970)] with modifications made for a square cross-section. For polymeric particles, we find that the mobility data agrees well over a large confinement range with the theory but under predicts for vesicles. The mobility of vesicles is higher in a square channel than in a circular tube, and does not depend significantly on membrane mechanical properties. The mobility of cancer cells is in good agreement with the theory up to λ ≈ 0.8, after which it deviates. Comparison of the mobility data of the three systems reveals that cancer cells have higher mobility than rigid particles but lower than vesicles, suggesting that the cell membrane frictional properties are in between a solid-like interface and a fluid bilayer. We explain further the differences in the mobility of the three systems by considering their shape deformation and surface flow on the interface. The results of this study may find potential applications in drug delivery and biomedical diagnostics.

Lipid partitioning at the nuclear envelope controls membrane biogenesis

PubMed Central

Barbosa, Antonio Daniel; Sembongi, Hiroshi; Su, Wen-Min; Abreu, Susana; Reggiori, Fulvio; Carman, George M.; Siniossoglou, Symeon

2015-01-01

Partitioning of lipid precursors between membranes and storage is crucial for cell growth, and its disruption underlies pathologies such as cancer, obesity, and type 2 diabetes. However, the mechanisms and signals that regulate this process are largely unknown. In yeast, lipid precursors are mainly used for phospholipid synthesis in nutrient-rich conditions in order to sustain rapid proliferation but are redirected to triacylglycerol (TAG) stored in lipid droplets during starvation. Here we investigate how cells reprogram lipid metabolism in the endoplasmic reticulum. We show that the conserved phosphatidate (PA) phosphatase Pah1, which generates diacylglycerol from PA, targets a nuclear membrane subdomain that is in contact with growing lipid droplets and mediates TAG synthesis. We find that cytosol acidification activates the master regulator of Pah1, the Nem1-Spo7 complex, thus linking Pah1 activity to cellular metabolic status. In the absence of TAG storage capacity, Pah1 still binds the nuclear membrane, but lipid precursors are redirected toward phospholipids, resulting in nuclear deformation and a proliferation of endoplasmic reticulum membrane. We propose that, in response to growth signals, activation of Pah1 at the nuclear envelope acts as a switch to control the balance between membrane biogenesis and lipid storage. PMID:26269581

Microscopic Analysis of Current and Mechanical Properties of Nafion® Studied by Atomic Force Microscopy

PubMed Central

Hiesgen, Renate; Helmly, Stefan; Galm, Ines; Morawietz, Tobias; Handl, Michael; Friedrich, K. Andreas

2012-01-01

The conductivity of fuel cell membranes as well as their mechanical properties at the nanometer scale were characterized using advanced tapping mode atomic force microscopy (AFM) techniques. AFM produces high-resolution images under continuous current flow of the conductive structure at the membrane surface and provides some insight into the bulk conducting network in Nafion membranes. The correlation of conductivity with other mechanical properties, such as adhesion force, deformation and stiffness, were simultaneously measured with the current and provided an indication of subsurface phase separations and phase distribution at the surface of the membrane. The distribution of conductive pores at the surface was identified by the formation of water droplets. A comparison of nanostructure models with high-resolution current images is discussed in detail. PMID:24958429

State diagram for adhesion dynamics of deformable capsules under shear flow.

PubMed

Luo, Zheng Yuan; Bai, Bo Feng

2016-08-17

Due to the significance of understanding the underlying mechanisms of cell adhesion in biological processes and cell capture in biomedical applications, we numerically investigate the adhesion dynamics of deformable capsules under shear flow by using a three-dimensional computational fluid dynamic model. This model is based on the coupling of the front tracking-finite element method for elastic mechanics of the capsule membrane and the adhesion kinetics simulation for adhesive interactions between capsules and functionalized surfaces. Using this model, three distinct adhesion dynamic states are predicted, such as detachment, rolling and firm-adhesion. Specifically, the effects of capsule deformability quantified by the capillary number on the transitions of these three dynamic states are investigated by developing an adhesion dynamic state diagram for the first time. At low capillary numbers (e.g. Ca < 0.0075), whole-capsule deformation confers the capsule a flattened bottom in contact with the functionalized surface, which hence promotes the rolling-to-firm-adhesion transition. It is consistent with the observations from previous studies that cell deformation promotes the adhesion of cells lying in the rolling regime. However, it is surprising to find that, at relatively high capillary numbers (e.g. 0.0075 < Ca < 0.0175), the effect of capsule deformability on its adhesion dynamics is far more complex than just promoting adhesion. High deformability of capsules makes their bottom take a concave shape with no adhesion bond formation in the middle. The appearance of this specific capsule shape inhibits the transitions of both rolling-to-firm-adhesion and detachment-to-rolling, and it means that capsule deformation no longer promotes the capsule adhesion. Besides, it is interesting to note that, when the capillary number exceeds a critical value (e.g. Ca = 0.0175), the rolling state no longer appears, since capsules exhibit large deviation from the spherical shape.

IFT88 influences chondrocyte actin organization and biomechanics.

PubMed

Wang, Z; Wann, A K T; Thompson, C L; Hassen, A; Wang, W; Knight, M M

2016-03-01

Primary cilia are microtubule based organelles which control a variety of signalling pathways important in cartilage development, health and disease. This study examines the role of the intraflagellar transport (IFT) protein, IFT88, in regulating fundamental actin organisation and mechanics in articular chondrocytes. The study used an established chondrocyte cell line with and without hypomorphic mutation of IFT88 (IFT88(orpk)). Confocal microscopy was used to quantify F-actin and myosin IIB organisation. Viscoelastic cell and actin cortex mechanics were determined using micropipette aspiration with actin dynamics visualised in live cells transfected with LifeACT-GFP. IFT88(orpk) cells exhibited a significant increase in acto-myosin stress fibre organisation relative to wild-type (WT) cells in monolayer and an altered response to cytochalasin D. Rounded IFT88(orpk) cells cultured in suspension exhibited reduced cortical actin expression with reduced cellular equilibrium modulus. Micropipette aspiration resulted in reduced membrane bleb formation in IFT88(orpk) cells. Following membrane blebbing, IFT88(orpk) cells exhibited slower reformation of the actin cortex. IFT88(orpk) cells showed increased actin deformability and reduced cortical tension confirming that IFT regulates actin cortex mechanics. The reduced cortical tension is also consistent with the reduced bleb formation. This study demonstrates for the first time that the ciliary protein IFT88 regulates fundamental actin organisation and the stiffness of the actin cortex leading to alterations in cell deformation, mechanical properties and blebbing in an IFT88 chondrocyte cell line. This adds to the growing understanding of the role of primary cilia and IFT in regulating cartilage biology. Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.

Impact of blood manufacturing and donor characteristics on membrane water permeability and in vitro quality parameters during hypothermic storage of red blood cells.

PubMed

Alshalani, Abdulrahman; Howell, Anita; Acker, Jason P

2018-02-01

Several factors have been proposed to influence the red blood cell storage lesion including storage duration, blood component manufacturing methodology, and donor characteristics [1,18]. The objectives of this study were to determine the impact of manufacturing method and donor characteristics on water permeability and membrane quality parameters. Red blood cell units were obtained from volunteer blood donors and grouped according to the manufacturing method and donor characteristics of sex and age. Membrane water permeability and membrane quality parameters, including deformability, hemolysis, osmotic fragility, hematologic indices, supernatant potassium, and supernatant sodium, were determined on day 5 ± 2, day 21, and day 42. Regression analysis was applied to evaluate the contribution of storage duration, manufacturing method, and donor characteristics on storage lesion. This study found that units processed using a whole blood filtration manufacturing method exhibited significantly higher membrane water permeability throughout storage compared to units manufactured using red cell filtration. Additionally, significant differences in hemolysis, supernatant potassium, and supernatant sodium were seen between manufacturing methods, however there were no significance differences between donor age and sex groups. Findings of this study suggest that the membrane-related storage lesion is initiated prior to the first day of storage with contributions by both blood manufacturing process and donor variability. The findings of this work highlight the importance of characterizing membrane water permeability during storage as it can be a predictor of the biophysical and chemical changes that affect the quality of stored red blood cells during hypothermic storage. Copyright © 2017 Elsevier Inc. All rights reserved.

Membrane Sculpting by F-BAR Domains Studied by Molecular Dynamics Simulations

PubMed Central

Yu, Hang; Schulten, Klaus

2013-01-01

Interplay between cellular membranes and their peripheral proteins drives many processes in eukaryotic cells. Proteins of the Bin/Amphiphysin/Rvs (BAR) domain family, in particular, play a role in cellular morphogenesis, for example curving planar membranes into tubular membranes. However, it is still unclear how F-BAR domain proteins act on membranes. Electron microscopy revealed that, in vitro, F-BAR proteins form regular lattices on cylindrically deformed membrane surfaces. Using all-atom and coarse-grained (CG) molecular dynamics simulations, we show that such lattices, indeed, induce tubes of observed radii. A 250 ns all-atom simulation reveals that F-BAR domain curves membranes via the so-called scaffolding mechanism. Plasticity of the F-BAR domain permits conformational change in response to membrane interaction, via partial unwinding of the domains 3-helix bundle structure. A CG simulation covering more than 350 µs provides a dynamic picture of membrane tubulation by lattices of F-BAR domains. A series of CG simulations identified the optimal lattice type for membrane sculpting, which matches closely the lattices seen through cryo-electron microscopy. PMID:23382665

Local mechanical response of cells to the controlled rotation of magnetic nanorods.

PubMed

Castillo, Matias; Ebensperger, Roberto; Wirtz, Denis; Walczak, Magdalena; Hurtado, Daniel E; Celedon, Alfredo

2014-11-01

The mechanical response of the cytoplasm was investigated by the intracellular implantation of magnetic nanorods and exposure to low-frequency rotatory magnetic fields. Nanorods (Pt-Ni, ∼200 nm diameter) fabricated by electrodeposition in templates of porous alumina with lengths of approximately 2 and 5 µm were inserted into NIH/3T3 fibroblasts and manipulated with a rotational magnetic field. Nanorod rotation was observed only for torques greater than 3.0 × 10(-16) Nm, suggesting a Bingham-type behavior of the cytoplasm. Higher torques produced considerable deformation of the intracellular material. The cell nucleus and cell membrane were significantly deformed by nanorods actuated by 4.5 × 10(-15) Nm torques. Our results demonstrate that nanorods under magnetic fields are an effective tool to mechanically probe the intracellular environment. We envision that our findings may contribute to the noninvasive and direct mechanical characterization of the cytoplasm. © 2014 Wiley Periodicals, Inc.

Local mechanical response of cells to the controlled rotation of magnetic nanorods

PubMed Central

Castillo, Matias; Ebensperger, Roberto; Wirtz, Denis; Walczak, Magdalena; Hurtado, Daniel E.; Celedon, Alfredo

2015-01-01

The mechanical response of the cytoplasm was investigated by the intracellular implantation of magnetic nanorods and exposure to low-frequency rotatory magnetic fields. Nanorods (Pt-Ni, ~200 nm diameter) fabricated by electrodeposition in templates of porous alumina with lengths of approximately 2 and 5 μm were inserted into NIH/ 3T3 fibroblasts and manipulated with a rotational magnetic field. Nanorod rotation was observed only for torques greater than 3.0 × 10−16 Nm, suggesting a Bingham-type behavior of the cytoplasm. Higher torques produced considerable deformation of the intracellular material. The cell nucleus and cell membrane were significantly deformed by nanorods actuated by 4.5 × 10−15 Nm torques. Our results demonstrate that nanorods under magnetic fields are an effective tool to mechanically probe the intracellular environment. We envision that our findings may contribute to the noninvasive and direct mechanical characterization of the cytoplasm. PMID:24700696

A multiphase approach to model ultrafiltration of deformable colloids

NASA Astrophysics Data System (ADS)

Haribabu, Malavika; Dunstan, Dave; Davidson, Malcolm; Harvie, Dalton

2017-11-01

Ultrafiltration (UF) is widely used in the dairy industry to fractionate and concentrate proteins, during the manufacture of milk protein concentrate and cheese. The protein build-up, comprising casein micelles (CM) and whey proteins, at the membrane surface during UF increases the resistance of the membrane system, thereby decreasing the performance of the process unit. CM have a complex structure that hydrodynamically behaves as a hard-sphere when dilute, but deforms beyond the random packing limit, forming a shear-thinning gel. This study employs a mixture model, based on the mixture phase continuity, Navier-Stokes equations, and solids continuity equation, to predict the solid concentration and velocity distribution during UF of CM. Micelle deformation is modelled as a function of volume fraction and dependent on its elastic modulus and particle size. The effect of deformation on gel permeability is implemented via Happel's permeability for hard spheres. Under crossflow conditions, the gel thickness is observed to increase along the membrane length, followed by a decrease towards the end of the membrane, resulting in an increase in flux at the latter section of the membrane. This study demonstrates that the membrane end-effects are important in determining UF performance.

Distributed sensing signal analysis of deformable plate/membrane mirrors

NASA Astrophysics Data System (ADS)

Lu, Yifan; Yue, Honghao; Deng, Zongquan; Tzou, Hornsen

2017-11-01

Deformable optical mirrors usually play key roles in aerospace and optical structural systems applied to space telescopes, radars, solar collectors, communication antennas, etc. Limited by the payload capacity of current launch vehicles, the deformable mirrors should be lightweight and are generally made of ultra-thin plates or even membranes. These plate/membrane mirrors are susceptible to external excitations and this may lead to surface inaccuracy and jeopardize relevant working performance. In order to investigate the modal vibration characteristics of the mirror, a piezoelectric layer is fully laminated on its non-reflective side to serve as sensors. The piezoelectric layer is segmented into infinitesimal elements so that microscopic distributed sensing signals can be explored. In this paper, the deformable mirror is modeled as a pre-tensioned plate and membrane respectively and sensing signal distributions of the two models are compared. Different pre-tensioning forces are also applied to reveal the tension effects on the mode shape and sensing signals of the mirror. Analytical results in this study could be used as guideline of optimal sensor/actuator placement for deformable space mirrors.

The plant cytoskeleton controls regulatory volume increase.

PubMed

Liu, Qiong; Qiao, Fei; Ismail, Ahmed; Chang, Xiaoli; Nick, Peter

2013-09-01

The ability to adjust cell volume is required for the adaptation to osmotic stress. Plant protoplasts can swell within seconds in response to hypoosmotic shock suggesting that membrane material is released from internal stores. Since the stability of plant membranes depends on submembraneous actin, we asked, whether this regulatory volume control depends on the cytoskeleton. As system we used two cell lines from grapevine which differ in their osmotic tolerance and observed that the cytoskeleton responded differently in these two cell lines. To quantify the ability for regulatory volume control, we used hydraulic conductivity (Lp) as readout and demonstrated a role of the cytoskeleton in protoplast swelling. Chelation of calcium, inhibition of calcium channels, or manipulation of membrane fluidity, did not significantly alter Lp, whereas direct manipulation of the cytoskeleton via specific chemical reagents, or indirectly, through the bacterial elicitor Harpin or activation of phospholipase D, was effective. By optochemical engineering of actin using a caged form of the phytohormone auxin we can break the symmetry of actin organisation resulting in a localised deformation of cell shape indicative of a locally increased Lp. We interpret our findings in terms of a model, where the submembraneous cytoskeleton controls the release of intracellular membrane stores during regulatory volume change. Copyright © 2013 Elsevier B.V. All rights reserved.

Computation of Static Shapes and Voltages for Micromachined Deformable Mirrors with Nonlinear Electrostatic Actuators

NASA Technical Reports Server (NTRS)

Wang, P. K. C.; Hadaegh, F. Y.

1996-01-01

In modeling micromachined deformable mirrors with electrostatic actuators whose gap spacings are of the same order of magnitude as those of the surface deformations, it is necessary to use nonlinear models for the actuators. In this paper, we consider micromachined deformable mirrors modeled by a membrane or plate equation with nonlinear electrostatic actuator characteristics. Numerical methods for computing the mirror deformation due to given actuator voltages and the actuator voltages required for producing the desired deformations at the actuator locations are presented. The application of the proposed methods to circular deformable mirrors whose surfaces are modeled by elastic membranes is discussed in detail. Numerical results are obtained for a typical circular micromachined mirror with electrostatic actuators.

A Wafer Transfer Technology for MEMS Adaptive Optics

NASA Technical Reports Server (NTRS)

Yang, Eui-Hyeok; Wiberg, Dean V.

2001-01-01

Adaptive optics systems require the combination of several advanced technologies such as precision optics, wavefront sensors, deformable mirrors, and lasers with high-speed control systems. The deformable mirror with a continuous membrane is a key component of these systems. This paper describes a new technique for transferring an entire wafer-level silicon membrane from one substrate to another. This technology is developed for the fabrication of a compact deformable mirror with a continuous facet. A 1 (mu)m thick silicon membrane, 100 mm in diameter, has been successfully transferred without using adhesives or polymers (i.e. wax, epoxy, or photoresist). Smaller or larger diameter membranes can also be transferred using this technique. The fabricated actuator membrane with an electrode gap of 1.5 (mu)m shows a vertical deflection of 0.37 (mu)m at 55 V.

Prediction of water loss and viscoelastic deformation of apple tissue using a multiscale model.

PubMed

Aregawi, Wondwosen A; Abera, Metadel K; Fanta, Solomon W; Verboven, Pieter; Nicolai, Bart

2014-11-19

A two-dimensional multiscale water transport and mechanical model was developed to predict the water loss and deformation of apple tissue (Malus × domestica Borkh. cv. 'Jonagold') during dehydration. At the macroscopic level, a continuum approach was used to construct a coupled water transport and mechanical model. Water transport in the tissue was simulated using a phenomenological approach using Fick's second law of diffusion. Mechanical deformation due to shrinkage was based on a structural mechanics model consisting of two parts: Yeoh strain energy functions to account for non-linearity and Maxwell's rheological model of visco-elasticity. Apparent parameters of the macroscale model were computed from a microscale model. The latter accounted for water exchange between different microscopic structures of the tissue (intercellular space, the cell wall network and cytoplasm) using transport laws with the water potential as the driving force for water exchange between different compartments of tissue. The microscale deformation mechanics were computed using a model where the cells were represented as a closed thin walled structure. The predicted apparent water transport properties of apple cortex tissue from the microscale model showed good agreement with the experimentally measured values. Deviations between calculated and measured mechanical properties of apple tissue were observed at strains larger than 3%, and were attributed to differences in water transport behavior between the experimental compression tests and the simulated dehydration-deformation behavior. Tissue dehydration and deformation in the high relative humidity range ( > 97% RH) could, however, be accurately predicted by the multiscale model. The multiscale model helped to understand the dynamics of the dehydration process and the importance of the different microstructural compartments (intercellular space, cell wall, membrane and cytoplasm) for water transport and mechanical deformation.

Imaging plasma membrane deformations with pTIRFM.

PubMed

Passmore, Daniel R; Rao, Tejeshwar C; Peleman, Andrew R; Anantharam, Arun

2014-04-02

To gain novel insights into the dynamics of exocytosis, our group focuses on the changes in lipid bilayer shape that must be precisely regulated during the fusion of vesicle and plasma membranes. These rapid and localized changes are achieved by dynamic interactions between lipids and specialized proteins that control membrane curvature. The absence of such interactions would not only have devastating consequences for vesicle fusion, but a host of other cellular functions that involve control of membrane shape. In recent years, the identity of a number of proteins with membrane-shaping properties has been determined. What remains missing is a roadmap of when, where, and how they act as fusion and content release progress. Our understanding of the molecular events that enable membrane remodeling has historically been limited by a lack of analytical methods that are sensitive to membrane curvature or have the temporal resolution to track rapid changes. PTIRFM satisfies both of these criteria. We discuss how pTIRFM is implemented to visualize and interpret rapid, submicron changes in the orientation of chromaffin cell membranes during dense core vesicle (DCV) fusion. The chromaffin cells we use are isolated from bovine adrenal glands. The membrane is stained with a lipophilic carbocyanine dye,1,1'-dioctadecyl-3,3,3',3'-tetramethylindodicarbocyanine, 4-chlorobenzenesulfonate, or diD. DiD intercalates in the membrane plane with a "fixed" orientation and is therefore sensitive to the polarization of the evanescent field. The diD-stained cell membrane is sequentially excited with orthogonal polarizations of a 561 nm laser (p-pol, s-pol). A 488 nm laser is used to visualize vesicle constituents and time the moment of fusion. Exocytosis is triggered by locally perfusing cells with a depolarizing KCl solution. Analysis is performed offline using custom-written software to understand how diD emission intensity changes relate to fusion pore dilation.

Aquaporin-0 Targets Interlocking Domains to Control the Integrity and Transparency of the Eye Lens

PubMed Central

Lo, Woo-Kuen; Biswas, Sondip K.; Brako, Lawrence; Shiels, Alan; Gu, Sumin; Jiang, Jean X.

2014-01-01

Purpose. Lens fiber cell membranes contain aquaporin-0 (AQP0), which constitutes approximately 50% of the total fiber cell membrane proteins and has a dual function as a water channel protein and an adhesion molecule. Fiber cell membranes also develop an elaborate interlocking system that is required for maintaining structural order, stability, and lens transparency. Herein, we used an AQP0-deficient mouse model to investigate an unconventional adhesion role of AQP0 in maintaining a normal structure of lens interlocking protrusions. Methods. The loss of AQP0 in AQP0−/− lens fibers was verified by Western blot and immunofluorescence analyses. Changes in membrane surface structures of wild-type and AQP0−/− lenses at age 3 to 12 weeks were examined with scanning electron microscopy. Preferential distribution of AQP0 in wild-type fiber cell membranes was analyzed with immunofluorescence and immunogold labeling using freeze-fracturing transmission electron microscopy. Results. Interlocking protrusions in young differentiating fiber cells developed normally but showed minor abnormalities at approximately 50 μm deep in the absence of AQP0 in all ages studied. Strikingly, protrusions in maturing fiber cells specifically underwent uncontrolled elongation, deformation, and fragmentation, while cells still retained their overall shape. Later in the process, these changes eventually resulted in fiber cell separation, breakdown, and cataract formation in the lens core. Immunolabeling at the light microscopy and transmission electron microscopy levels demonstrated that AQP0 was particularly enriched in interlocking protrusions in wild-type lenses. Conclusions. This study suggests that AQP0 exerts its primary adhesion or suppression role specifically to maintain the normal structure of interlocking protrusions that is critical to the integrity and transparency of the lens. PMID:24458158

Double-layered cell transfer technology for bone regeneration

PubMed Central

Akazawa, Keiko; Iwasaki, Kengo; Nagata, Mizuki; Yokoyama, Naoki; Ayame, Hirohito; Yamaki, Kazumasa; Tanaka, Yuichi; Honda, Izumi; Morioka, Chikako; Kimura, Tsuyoshi; Komaki, Motohiro; Kishida, Akio; Izumi, Yuichi; Morita, Ikuo

2016-01-01

For cell-based medicine, to mimic in vivo cellular localization, various tissue engineering approaches have been studied to obtain a desirable arrangement of cells on scaffold materials. We have developed a novel method of cell manipulation called “cell transfer technology”, enabling the transfer of cultured cells onto scaffold materials, and controlling cell topology. Here we show that using this technique, two different cell types can be transferred onto a scaffold surface as stable double layers or in patterned arrangements. Various combinations of adherent cells were transferred to a scaffold, amniotic membrane, in overlapping bilayers (double-layered cell transfer), and transferred cells showed stability upon deformations of the material including folding and trimming. Transplantation of mesenchymal stem cells from periodontal ligaments (PDLSC) and osteoblasts, using double-layered cell transfer significantly enhanced bone formation, when compared to single cell type transplantation. Our findings suggest that this double-layer cell transfer is useful to produce a cell transplantation material that can bear two cell layers. Moreover, the transplantation of an amniotic membrane with PDLSCs/osteoblasts by cell transfer technology has therapeutic potential for bone defects. We conclude that cell transfer technology provides a novel and unique cell transplantation method for bone regeneration. PMID:27624174

Unraveling supported lipid bilayer formation kinetics: osmotic effects.

PubMed

Hain, Nicole; Gallego, Marta; Reviakine, Ilya

2013-02-19

Solid-supported lipid bilayers are used as cell membrane models and form the basis of biomimetic and biosensor platforms. The mechanism of their formation from adsorbed liposomes is not well-understood. Using membrane-permeable solute glycerol, impermeable solutes sucrose and dextran, and a pore forming peptide melittin, we studied experimentally how osmotic effects affect the kinetics of the adsorbed liposome-to-bilayer transition. We find that its rate is enhanced if adsorbed liposomes are made permeable but is not significantly retarded by impermeable solutes. The results are explained in terms of adsorbed liposome deformation and formation of transmembrane pores.

Passive and active response of bacteria under mechanical compression

NASA Astrophysics Data System (ADS)

Garces, Renata; Miller, Samantha; Schmidt, Christoph F.; Byophysics Team; Institute of Medical Sciences Collaboration

Bacteria display simple but fascinating cellular structures and geometries. Their shapes are the result of the interplay between osmotic pressure and cell wall construction. Typically, bacteria maintain a high difference of osmotic pressure (on the order of 1 atm) to the environment. This pressure difference (turgor pressure) is supported by the cell envelope, a composite of lipid membranes and a rigid cell wall. The response of the cell envelope to mechanical perturbations such as geometrical confinements is important for the cells survival. Another key property of bacteria is the ability to regulate turgor pressure after abrupt changes of external osmotic conditions. This response relies on the activity of mechanosensitive (MS) channels: membrane proteins that release solutes in response to excessive stress in the cell envelope. We here present experimental data on the mechanical response of the cell envelope and on turgor regulation of bacteria subjected to compressive forces. We indent living cells with micron-sized beads attached to the cantilever of an atomic force microscope (AFM). This approach ensures global deformation of the cell. We show that such mechanical loading is sufficient to gate mechanosensitive channels in isosmotic conditions.

Lattice preferred orientation in MnGeO3 post-perovskite at high-temperature

NASA Astrophysics Data System (ADS)

Nagaya, Y.; Hirose, K.; Sata, N.; Ohishi, Y.

2009-12-01

In the Earth’s lowermost mantle which is called D” layer, shear-wave splitting is often observed. The velocity of horizontally polarized S-waves are faster than polarized S-waves in many areas of the D” layer. The D” layer is now recognized as being made up with the post-perovskite (PPv)-type MgSiO3 phase. MgSiO3 PPv has a strong elastic anisotropy because of its layered crystal structure. Therefore, it is expected that a lattice preferred orientation (LPO) of PPv may explain the observed seismic anisotropy. LPOs of PPv have been investigated by the high-pressure experiments using a diamond anvil cell (DAC) (Merkel et al., 2006; 2007; Okada et al., 2009). However, the reported experiments using the DAC were made only at the room temperature. In order to understand the nature of PPv deformation under the lower mantle conditions, it is necessary to operate the deformation experiments at high-temperature (~2500 K). In this study, so as to examine the LPO and the dominant slip plane of PPv at simultaneously high P-T conditions, we conducted the high-temperature plastic deformation experiments in a laser-heated diamond anvil cell (LHDAC) using synchrotron radial X-ray diffraction techniques at the beamline BL10XU, SPring-8. In the radial X-ray diffraction experiments, X-ray was irradiated to the sample perpendicular to the compression axis through gasket. LPO was investigated on the basis of the variations of diffraction intensity. We adopted a cubic boron nitride and beryllium composite gasket to obtain a radial X-ray diffraction pattern. In order to deform a sample at high temperature, we had newly developed a membrane system for the deformation experiments. We are able to regulate the gas pressure in the membrane of the DAC, and therefore compress the sample at high temperature during the laser heating. Starting material was orthopyroxene (OPx) with a composition of MnGeO3, which is an analogue of MgSiO3. First, MnGeO3 PPv was synthesized directly from OPx around 60 GPa in the LHDAC. Subsequently, PPv was plastically deformed by further compression at high-temperature during the laser heating. We also conducted the room-temperature deformation experiments. We will discuss the deformation mechanism of the PPv at high P-T conditions.

Large Deformation and Adhesive Contact Studies of Axisymmetric Membranes

PubMed Central

Laprade, Evan J.; Long, Rong; Pham, Jonathan; Lawrence, Jimmy; Emrick, Todd; Crosby, Alfred; Hui, Chung-Yuen; Shull, Kenneth R.

2013-01-01

A model membrane contact system consisting of an acrylic copolymer membrane and polydimethyl-siloxane substrate was utilized to evaluate a recently developed nonlinear large-deformation adhesive contact analysis. Direct measurements of the local membrane apex strain during non-contact inflation indicated that the neo-Hookean model provides an accurate measure of membrane strain and supports its use as the strain energy function for the analysis. A time dependent modulus emerges from the analysis, with principal tensions obtained from a comparison of predicted and experimental membrane profiles. A displacement controlled geometry was more easily modeled than the pressure controlled geometry, the applicability of the analysis was limited by wrinkling instabilities. The substantial viscoelastic behavior of these membranes made it difficult to describe the entire membrane with a single modulus, given the nonuniform deformation history of the membranes. Given the difficulty in determining membrane tension from the measured pressure and profile fits using the model, the peel energy was used as a simpler measure of adhesion. Using an analytical balance in the displacement controlled geometry, the membrane tension at the contact line was directly measured. Coupled with contact angle imaging, the peel energy was determined. For the model membranes studied, this peel energy described the membrane/substrate adhesive interactions quite well, giving well-defined peel energies that were independent of the detailed strain state of the membrane. PMID:23289644

Integrated optical design for highly dynamic laser beam shaping with membrane deformable mirrors

NASA Astrophysics Data System (ADS)

Pütsch, Oliver; Stollenwerk, Jochen; Loosen, Peter

2017-02-01

The utilization of membrane deformable mirrors has raised its importance in laser materials processing since they enable the generation of highly spatial and temporal dynamic intensity distributions for a wide field of applications. To take full advantage of these devices for beam shaping, the huge amount of degrees of freedom has to be considered and optimized already within the early stage of the optical design. Since the functionality of commercial available ray-tracing software has been mainly specialized on geometric dependencies and their optimization within constraints, the complex system characteristics of deformable mirrors cannot be sufficiently taken into account yet. The main reasons are the electromechanical interdependencies of electrostatic membrane deformable mirrors, namely saturation and mechanical clamping, that result in non-linear deformation. This motivates the development of an integrative design methodology. The functionality of the ray-tracing program ZEMAX is extended with a model of an electrostatic membrane mirror. This model is based on experimentally determined influence functions. Furthermore, software routines are derived and integrated that allow for the compilation of optimization criteria for the most relevant analytically describable beam shaping problems. In this way, internal optimization routines can be applied for computing the appropriate membrane deflection of the deformable mirror as well as for the parametrization of static optical components. The experimental verification of simulated intensity distributions demonstrates that the beam shaping properties can be predicted with a high degree of reliability and precision.

Design of a new membrane stretching device

NASA Astrophysics Data System (ADS)

Shao, Yiran

Cell stretching device has been applied into the lab use for many years to help researchers study about the behavior of cells during the stretching process. Because the cell responses to the different mechanical stimuli, especially in the case of disease, the cell stretching device is a necessary tool to study the cell behavior in a controlled environment. However existing devices have limitations, such as too big to fit the culture chamber, unable to be observed during the stretching process and too expensive to fabricate. In this thesis, a new cell stretcher is designed to resolve these limitations. Many typical cell stretching devices only work under simple conditions. For instance they can only apply the strain on the cell in uniaxial or equibiaxial directions. On the other hand the environment of cells' survival is varying. Many new cell stretchers have been developed, which have the same property that cells can be stretched via the radical deformation of the elastomeric membrane. The aim of this new design is to create a cell stretching device that fits in general lab conditions. This device is designed to fit on a microscope to observe, as well as in the incubator. In addition, two small step motors are used to control the strain, adjust the frequency, and maintain the stability precisely. Problems such as the culture media leakage and the membrane breakage are solved by the usage of multiple materials for both the cell stretcher and the membrane. Based on the experimental results, this device can satisfy the requirements of target users with a reduced manufacturing cost. In the future, an auto-focus tracking function will be developed to allow real time observation of the cells' behavior.

2-D Model for Normal and Sickle Cell Blood Microcirculation

NASA Astrophysics Data System (ADS)

Tekleab, Yonatan; Harris, Wesley

2011-11-01

Sickle cell disease (SCD) is a genetic disorder that alters the red blood cell (RBC) structure and function such that hemoglobin (Hb) cannot effectively bind and release oxygen. Previous computational models have been designed to study the microcirculation for insight into blood disorders such as SCD. Our novel 2-D computational model represents a fast, time efficient method developed to analyze flow dynamics, O2 diffusion, and cell deformation in the microcirculation. The model uses a finite difference, Crank-Nicholson scheme to compute the flow and O2 concentration, and the level set computational method to advect the RBC membrane on a staggered grid. Several sets of initial and boundary conditions were tested. Simulation data indicate a few parameters to be significant in the perturbation of the blood flow and O2 concentration profiles. Specifically, the Hill coefficient, arterial O2 partial pressure, O2 partial pressure at 50% Hb saturation, and cell membrane stiffness are significant factors. Results were found to be consistent with those of Le Floch [2010] and Secomb [2006].

Experimental characterization of mm-wave detection by a micro-array of Golay cells

NASA Astrophysics Data System (ADS)

Denison, Douglas R.; Knotts, Michael E.; McConney, Michael E.; Tsukruk, Vladimir V.

2009-05-01

We present experimental results for an uncooled imaging focal plane array technology that consists of a polymer/metal/polymer layered membrane suspended over a micro-fabricated array of cavities. The device operation is Golay-like (heating of air in the cavity causes a detectable deflection of the membrane proportional to incident EM power), but potentially offers both greater sensitivity and more read-out options (optical or electrical) than a traditional Golay cell through tailoring of the membrane properties. The membrane is formed from a layer-by-layer deposition of polymer with one or more monolayers of gold nanoparticles (or other metal) that help control the membrane's elasticity and deformation-dependent optical reflectivity/electrical conductivity. Baseline capabilities of the device have been established through optical measurements of membrane deflection due to incident mm-wave radiation modulated at 30 Hz (corresponding to a video refresh rate). The device demonstrates an NEP of 300 nW/√Hz at 105 GHz for a 19-layer membrane (9 poly/1 Au/9 poly) suspended over an array of 80 μm diameter cavities (depth = 100 μm) etched in a 500 μm thick substrate of Si. Calculations of membrane sensitivity show that this NEP could be reduced to ~ 100 pW/√Hz with enlarged cavity diameters on the order of 600 μm.

Simulations of the erythrocyte cytoskeleton at large deformation. II. Micropipette aspiration.

PubMed Central

Discher, D E; Boal, D H; Boey, S K

1998-01-01

Coarse-grained molecular models of the erythrocyte membrane's spectrin cytoskeleton are presented in Monte Carlo simulations of whole cells in micropipette aspiration. The nonlinear chain elasticity and sterics revealed in more microscopic cytoskeleton models (developed in a companion paper; Boey et al., 1998. Biophys. J. 75:1573-1583) are faithfully represented here by two- and three-body effective potentials. The number of degrees of freedom of the system are thereby reduced to a range that is computationally tractable. Three effective models for the triangulated cytoskeleton are developed: two models in which the cytoskeleton is stress-free and does or does not have internal attractive interactions, and a third model in which the cytoskeleton is prestressed in situ. These are employed in direct, finite-temperature simulations of erythrocyte deformation in a micropipette. All three models show reasonable agreement with aspiration measurements made on flaccid human erythrocytes, but the prestressed model alone yields optimal agreement with fluorescence imaging experiments. Ensemble-averaging of nonaxisymmetrical, deformed structures exhibiting anisotropic strain are thus shown to provide an answer to the basic question of how a triangulated mesh such as that of the red cell cytoskeleton deforms in experiment. PMID:9726959

Simulations of the erythrocyte cytoskeleton at large deformation. II. Micropipette aspiration.

PubMed

Discher, D E; Boal, D H; Boey, S K

1998-09-01

Coarse-grained molecular models of the erythrocyte membrane's spectrin cytoskeleton are presented in Monte Carlo simulations of whole cells in micropipette aspiration. The nonlinear chain elasticity and sterics revealed in more microscopic cytoskeleton models (developed in a companion paper; Boey et al., 1998. Biophys. J. 75:1573-1583) are faithfully represented here by two- and three-body effective potentials. The number of degrees of freedom of the system are thereby reduced to a range that is computationally tractable. Three effective models for the triangulated cytoskeleton are developed: two models in which the cytoskeleton is stress-free and does or does not have internal attractive interactions, and a third model in which the cytoskeleton is prestressed in situ. These are employed in direct, finite-temperature simulations of erythrocyte deformation in a micropipette. All three models show reasonable agreement with aspiration measurements made on flaccid human erythrocytes, but the prestressed model alone yields optimal agreement with fluorescence imaging experiments. Ensemble-averaging of nonaxisymmetrical, deformed structures exhibiting anisotropic strain are thus shown to provide an answer to the basic question of how a triangulated mesh such as that of the red cell cytoskeleton deforms in experiment.

Interaction pathways between soft lipid nanodiscs and plasma membranes: A molecular modeling study.

PubMed

Li, Shixin; Luo, Zhen; Xu, Yan; Ren, Hao; Deng, Li; Zhang, Xianren; Huang, Fang; Yue, Tongtao

2017-10-01

Lipid nanodisc, a model membrane platform originally synthesized for study of membrane proteins, has recently been used as the carrier to deliver amphiphilic drugs into target tumor cells. However, the central question of how cells interact with such emerging nanomaterials remains unclear and deserves our research for both improving the delivery efficiency and reducing the side effect. In this work, a binary lipid nanodisc is designed as the minimum model to investigate its interactions with plasma membranes by using the dissipative particle dynamics method. Three typical interaction pathways, including the membrane attachment with lipid domain exchange of nanodiscs, the partial membrane wrapping with nanodisc vesiculation, and the receptor-mediated endocytosis, are discovered. For the first pathway, the boundary normal lipids acting as ligands diffuse along the nanodisc rim to gather at the membrane interface, repelling the central bola lipids to reach a stable membrane attachment. If bola lipids are positioned at the periphery and act as ligands, they diffuse to form a large aggregate being wrapped by the membrane, leaving the normal lipids exposed on the membrane exterior by assembling into a vesicle. Finally, by setting both central normal lipids and boundary bola lipids as ligands, the receptor-mediated endocytosis occurs via both deformation and self-rotation of the nanodiscs. All above pathways for soft lipid nanodiscs are quite different from those for rigid nanoparticles, which may provide useful guidelines for design of soft lipid nanodiscs in widespread biomedical applications. Copyright © 2017 Elsevier B.V. All rights reserved.

Novel electrospun gas diffusion layers for polymer electrolyte membrane fuel cells: Part II. In operando synchrotron imaging for microscale liquid water transport characterization

NASA Astrophysics Data System (ADS)

Chevalier, S.; Ge, N.; Lee, J.; George, M. G.; Liu, H.; Shrestha, P.; Muirhead, D.; Lavielle, N.; Hatton, B. D.; Bazylak, A.

2017-06-01

This is the second paper in a two-part series in which we investigate the impact of the gas diffusion layer structure on the liquid water distribution in an operating polymer electrolyte membrane (PEM) fuel cell through the procedures of design, fabrication, and testing of novel hydrophobic electrospun gas diffusion layers (eGDLs). In this work, fibre diameters and alignment in eGDLs are precisely controlled, and concurrent synchrotron X-ray radiography and electrochemical impedance spectroscopy (EIS) are used to evaluate the influence of the controlled eGDL parameters on the liquid water distribution and on membrane liquid water content. For eGDLs with small fibre diameters (150-200 nm) and correspondingly smaller pore sizes, reduced liquid water accumulation under the flow field ribs is observed. However, more liquid water is pinned onto the eGDL - at the interface with flow field channels. Orienting fibre alignment perpendicular to the flow field channel direction leads to improved eGDL-catalyst layer contact and prevents rib-channel membrane deformation. On the other hand, eGDLs facilitate significant membrane dry-out, even under highly humidified operating conditions at high current densities.

Deformations of a pre-stretched elastic membrane driven by non-uniform electroosmotic flow

NASA Astrophysics Data System (ADS)

Bercovici, Moran; Boyko, Evgeniy; Gat, Amir

2016-11-01

We study viscous-elastic dynamics of fluid confined between a rigid plate and a pre-stretched elastic membrane subjected to non-uniform electroosmotic flow, and focus on the case of a finite-size membrane clamped at its boundaries. Considering small deformations of a strongly pre-stretched membrane, and applying the lubrication approximation for the flow, we derive a linearized leading-order non-homogenous 4th order diffusion equation governing the deformation and pressure fields. We derive a time-dependent Green's function for a rectangular domain, and use it to obtain several basic solutions for the cases of constant and time varying electric fields. In addition, defining an asymptotic expansion where the small parameter is the ratio of the induced to prescribed tension, we obtain a set of four one-way coupled equations providing a first order correction for the deformation field. Funded by the European Research Council (ERC) under the Horizon 2020 Research and Innovation Programme, Grant agreement No. 678734 (MetamorphChip).

The Effect of Sepsis on the Erythrocyte

PubMed Central

Bateman, Ryon M.; Sharpe, Michael D.; Singer, Mervyn; Ellis, Christopher G.

2017-01-01

Sepsis induces a wide range of effects on the red blood cell (RBC). Some of the effects including altered metabolism and decreased 2,3-bisphosphoglycerate are preventable with appropriate treatment, whereas others, including decreased erythrocyte deformability and redistribution of membrane phospholipids, appear to be permanent, and factors in RBC clearance. Here, we review the effects of sepsis on the erythrocyte, including changes in RBC volume, metabolism and hemoglobin’s affinity for oxygen, morphology, RBC deformability (an early indicator of sepsis), antioxidant status, intracellular Ca2+ homeostasis, membrane proteins, membrane phospholipid redistribution, clearance and RBC O2-dependent adenosine triphosphate efflux (an RBC hypoxia signaling mechanism involved in microvascular autoregulation). We also consider the causes of these effects by host mediated oxidant stress and bacterial virulence factors. Additionally, we consider the altered erythrocyte microenvironment due to sepsis induced microvascular dysregulation and speculate on the possible effects of RBC autoxidation. In future, a better understanding of the mechanisms involved in sepsis induced erythrocyte pathophysiology and clearance may guide improved sepsis treatments. Evidence that small molecule antioxidants protect the erythrocyte from loss of deformability, and more importantly improve septic patient outcome suggest further research in this area is warranted. While not generally considered a critical factor in sepsis, erythrocytes (and especially a smaller subpopulation) appear to be highly susceptible to sepsis induced injury, provide an early warning signal of sepsis and are a factor in the microvascular dysfunction that has been associated with organ dysfunction. PMID:28885563

Disruption of 3D MCF-12A Breast Cell Cultures by Estrogens – An In Vitro Model for ER-Mediated Changes Indicative of Hormonal Carcinogenesis

PubMed Central

Marchese, Stephanie; Silva, Elisabete

2012-01-01

Introduction Estrogens regulate the proliferation of normal and neoplastic breast epithelium. Although the intracellular mechanisms of estrogens in the breast are largely understood, little is known about how they induce changes in the structure of the mammary epithelium, which are characteristic of breast cancer. In vitro three dimensional (3D) cultures of immortalised breast epithelial cells recapitulate features of the breast epithelium in vivo, including formation of growth arrested acini with hollow lumen and basement membrane. This model can also reproduce features of malignant transformation and breast cancer, such as increased cellular proliferation and filling of the lumen. However, a system where a connection between estrogen receptor (ER) activation and disruption of acini formation can be studied to elucidate the role of estrogens is still missing. Methods/Principal Findings We describe an in vitro 3D model for breast glandular structure development, using breast epithelial MCF-12A cells cultured in a reconstituted basement membrane matrix. These cells are estrogen receptor (ER)α, ERβ and G-protein coupled estrogen receptor 1 (GPER) competent, allowing the investigation of the effects of estrogens on mammary gland formation and disruption. Under normal conditions, MCF-12A cells formed organised acini, with deposition of basement membrane and hollow lumen. However, treatment with 17β-estradiol, and the exogenous estrogens bisphenol A and propylparaben resulted in deformed acini and filling of the acinar lumen. When these chemicals were combined with ER and GPER inhibitors (ICI 182,780 and G-15, respectively), the deformed acini recovered normal features, such as a spheroid shape, proliferative arrest and luminal clearing, suggesting a role for the ER and GPER in the estrogenic disruption of acinar formation. Conclusion This new model offers the opportunity to better understand the role of the ER and GPER in the morphogenesis of breast glandular structure as well as the events implicated in breast cancer initiation and progression. PMID:23056216

Red blood cell membrane water permeability increases with length of ex vivo storage.

PubMed

Alshalani, Abdulrahman; Acker, Jason P

2017-06-01

Water transport across the red blood cell (RBC) membrane is an essential cell function that needs to be preserved during ex vivo storage. Progressive biochemical depletion during storage can result in significant conformational and compositional changes to the membrane. Characterizing the changes to RBC water permeability can help in evaluating the quality of stored blood products and aid in the development of improved methods for the cryopreservation of red blood cells. This study aimed to characterize the water permeability (L p ), osmotically inactive fraction (b), and Arrhenius activation energy (E a ) at defined storage time-points throughout storage and to correlate the observed results with other in vitro RBC quality parameters. RBCs were collected from age- and sex-matched blood donors. A stopped flow spectrophotometer was used to determine L p and b by monitoring changes in hemoglobin autofluorescence when RBCs were exposed to anisotonic solutions. Experimental values of L p were characterized at three different temperatures (4, 20 and 37 °C) to determine the E a . Results showed that L p , b, and E a of stored RBCs significantly increase by day 21 of storage. Degradation of the RBC membrane with length of storage was seen as an increase in hemolysis and supernatant potassium, and a decrease in deformability, mean corpuscular hemoglobin concentration and supernatant sodium. RBC osmotic characteristics were shown to change with storage and correlate with changes in RBC membrane quality metrics. Monitoring water parameters is a predictor of membrane damage and loss of membrane integrity in ex vivo stored RBCs. Copyright © 2017 Elsevier Inc. All rights reserved.

Protective effects of a freeze-dried extract of vegetables and fruits on the hydroxyl radical-mediated oxidative damage of DNA and decrease of erythrocytes deformability.

PubMed

Wang, Hsiao-Ning; Liu, Tsan-Zon; Chen, Ya-Lei; Shiuan, David

2007-01-01

The protective effects of a freeze-dried extracts of vegetables and fruits (BauYuan; BY) on the hydroxyl radical-mediated DNA strand breakages and the structural integrity of human red blood cells (RBCs) were investigated. First, the supercoiled plasmid (pEGFP-C1) DNA was subjected to oxidation damage by an ascorbate-fortified Fenton reaction and the protective effects were analyzed by agarose gel electrophoresis. In the absence of BY extracts, exposure of the high-throughput .OH-generating system (Fe2+ concentration >1.0 microM) caused a complete fragmentation of DNA. Supplementation of BY extract (1 mg/mL) to the plasmid DNA prior to the exposure could prevent it significantly. In contrast, as the plasmid exposed to a low-grade .OH-generating system (Fe2+<0.1 microM), the BY extract (1 mg/mL) provided an almost complete protection. Next, the cell deformabilities were measured to assess the protection effects of various BY extracts on human erythrocytes exposed to the oxidative insults. We found that both the aqueous extract and the organic solvent-derived extracts could strongly protect human RBCs from the reactive oxygen species (ROS)-mediated decrease in the deformability indices. The results implicated that the BY extracts could effectively protect the cell membrane integrity via scavenging ROS which enabling RBCs to maintain a balance of water content and surface area to prevent the drop of cell deformability.

Uncoupling Store-Operated Ca2+ Entry and Altered Ca2+ Release from Sarcoplasmic Reticulum through Silencing of Junctophilin Genes

PubMed Central

Hirata, Yutaka; Brotto, Marco; Weisleder, Noah; Chu, Yi; Lin, Peihui; Zhao, Xiaoli; Thornton, Angela; Komazaki, Shinji; Takeshima, Hiroshi; Ma, Jianjie; Pan, Zui

2006-01-01

Junctophilin (JP) mediates the close contact between cell surface and intracellular membranes in muscle cells ensuring efficient excitation-contraction coupling. Here we demonstrate that disruption of triad junction structure formed by the transverse tubular (TT) invagination of plasma membrane and terminal cisternae of sarcoplasmic reticulum (SR) by reduction of JP expression leads to defective Ca2+ homeostasis in muscle cells. Using adenovirus with small hairpin interference RNA (shRNA) against both JP1 and JP2 genes, we could achieve acute suppression of JPs in skeletal muscle fibers. The shRNA-treated muscles exhibit deformed triad junctions and reduced store-operated Ca2+ entry (SOCE), which is likely due to uncoupled retrograde signaling from SR to TT. Knockdown of JP also causes a reduction in SR Ca2+ storage and altered caffeine-induced Ca2+ release, suggesting an orthograde regulation of the TT membrane on the SR Ca2+ release machinery. Our data demonstrate that JPs play an important role in controlling overall intracellular Ca2+ homeostasis in muscle cells. We speculate that altered expression of JPs may underlie some of the phenotypic changes associated with certain muscle diseases and aging. PMID:16565048

Connectingthe puzzle pieces between cytoskeleton andsecretory pathway

PubMed Central

Gurel, Pinar S.; Hatch, Anna L.; Higgs, Henry N.

2014-01-01

A tendency in cell biology is to divide and conquer. For example, decades of painstaking work have led to an understanding of endoplasmic reticulum (ER) and Golgi structure, dynamics, and transport. In parallel, cytoskeletal researchers have revealed a fantastic diversity of structure and cellular function in both actin and microtubules. Increasingly, these areas overlap, necessitating an understanding of both organelle and cytoskeletal biology. This review addressesconnections between the actin/microtubule cytoskeletons and organelles in animal cells, focusing on threetopics: ER structure/function, ER-to-Golgi transport; and Golgi structure/function. Making these connections has been challenging, due to 1) the small sizes and dynamic characteristics of some components, 2) the fact that organelle-specific cytoskeleton can easily be obscured by more abundant cytoskeletal structures, and 3) the difficulties in imaging membranes and cytoskeleton simultaneously, especially at the ultra-structural level. One major concept is that the cytoskeleton is frequently used to generate force for membrane movement, with two potential consequences: translocation of the organelle, or deformation of the organelle membrane. While initially discussing issues common to metazoan cells in general, we subsequently highlight specific features of neurons, since these highly polarized cells present unique challenges for organellar distribution and dynamics. PMID:25050967

Influence of surface tension on the instabilities and bifurcations of a particle in a drop under shear

NASA Astrophysics Data System (ADS)

Gallaire, Francois; Zhu, Lailai

2016-11-01

While the deformation regimes under flow of anuclear cells, like red blood cells, have been widely analyzed, the dynamics of nuclear cells are less explored. The objective of this work is to investigate the interplay between the stiff nucleus, modeled here as a rigid spherical particle and the surrounding deformable cell membrane, modeled for simplicity as an immiscible droplet, subjected to an external unbounded plane shear flow. A three-dimensional boundary integral implementation is developed to describe the interface-structure interaction characterized by two dimensionless numbers: the capillary number Ca , defined as the ratio of shear to capillary forces and and the particle-droplet size ratio. For large Ca , i.e. very deformable droplets, the particle has a stable equilibrium position at the center of the droplet. However, for smaller Ca , both the plane symmetry and the time invariance are broken and the particle migrates to a closed orbit located off the symmetry plane, reaching a limit cycle. For even smaller capillary numbers, the time invariance is restored and the particle reaches a steady equilibrium position off the symmetry plane. This series of bifurcations is analyzed and possible physical mechanisms from which they originate are discussed. Financial support by ERC Grant SimCoMiCs 280117 is gratefully acknowledged.

Fluid Mechanics of the Vascular Basement Membrane in the Brain

NASA Astrophysics Data System (ADS)

Coloma, Mikhail; Hui, Jonathan; Chiarot, Paul; Huang, Peter; Carare, Roxana; McLeod, Kenneth; Schaffer, David

2013-11-01

Beta-amyloid is a normal product of brain metabolic function and is found within the interstitial fluid of the brain. Failure of the clearance of beta-amyloid from the aging brain leads to its accumulation within the walls of arteries and to Alzheimer's disease. The vascular basement membrane (VBM) within the walls of cerebral arteries surrounds the spirally arranged smooth muscle cells and represents an essential pathway for removal of beta-amyloid from the brain. This process fails with the stiffening of arterial walls associated with aging. In this study we hypothesize that the deformation of the VBM associated with arterial pulsations drives the interstitial fluid to drain in the direction opposite of the arterial blood flow. This hypothesis is theoretically investigated by modeling the VBM as a thin, coaxial, fluid-filled porous medium surrounding a periodically deforming cylindrical tube. Flow and boundary conditions required to achieve such a backward clearance are derived through a control volume analysis of mass, momentum, and energy.

Three-dimensional refractive index tomograms and deformability of individual human red blood cells from cord blood of newborn infants and maternal blood.

PubMed

Park, HyunJoo; Ahn, Taegyu; Kim, Kyoohyun; Lee, Sangyun; Kook, Song-Yi; Lee, Dongheon; Suh, In Bum; Na, Sunghun; Park, YongKeun

2015-01-01

Red blood cells (RBCs) from the cord blood of newborn infants have distinctive functions in fetal and infant development. To systematically investigate the biophysical characteristics of individual cord RBCs in newborn infants, a comparative study was performed on RBCs from the cord blood of newborn infants and from adult mothers or nonpregnant women using optical holographic microtomography. Optical measurements of the distributions of the three-dimensional refractive indices and the dynamic membrane fluctuations of individual RBCs were used to investigate the morphological, biochemical, and mechanical properties of cord, maternal, and adult RBCs at the individual cell level. The volume and surface area of the cord RBCs were significantly larger than those of the RBCs from nonpregnant women, and the cord RBCs had more flattened shapes than that of the RBCs in adults. In addition, the hemoglobin (Hb) content in the cord RBCs from newborns was significantly higher. The Hb concentration in the cord RBCs was higher than that in the nonpregnant women or maternal RBCs, but they were within the physiological range of adults. Interestingly, the amplitudes of the dynamic membrane fluctuations in cord RBCs were comparable to those in nonpregnant women and maternal RBCs, suggesting that the deformability of cord RBCs is similar to that of healthy RBCs in adults.

Confined semiflexible polymers suppress fluctuations of soft membrane tubes.

PubMed

Mirzaeifard, Sina; Abel, Steven M

2016-02-14

We use Monte Carlo computer simulations to investigate tubular membrane structures with and without semiflexible polymers confined inside. At small values of membrane bending rigidity, empty fluid and non-fluid membrane tubes exhibit markedly different behavior, with fluid membranes adopting irregular, highly fluctuating shapes and non-fluid membranes maintaining extended tube-like structures. Fluid membranes, unlike non-fluid membranes, exhibit a local maximum in specific heat as their bending rigidity increases. The peak is coincident with a transition to extended tube-like structures. We further find that confining a semiflexible polymer within a fluid membrane tube reduces the specific heat of the membrane, which is a consequence of suppressed membrane shape fluctuations. Polymers with a sufficiently large persistence length can significantly deform the membrane tube, with long polymers leading to localized bulges in the membrane that accommodate regions in which the polymer forms loops. Analytical calculations of the energies of idealized polymer-membrane configurations provide additional insight into the formation of polymer-induced membrane deformations.

[Quantitative changes of main components of erythrocyte membranes which define architectonics of cells under pttg gene knockout].

PubMed

Kaniuka, O P; Filiak, Ie Z; Kulachkovs'kyĭ, O R; Osyp, Iu L; Sybirna, N O

2014-01-01

A pttg gene knockout affects the functional state of erythron in mice which could be associated with structural changes in the structure of erythrocyte membranes. The pttg gene knockout causes a significant modification of fatty acids composition of erythrocyte membrane lipids by reducing the content of palmitic acid and increasing of polyunsaturated fatty acids amount by 18%. Analyzing the erythrocyte surface architectonics of mice under pttg gene knockout, it was found that on the background of reduction of the functionally complete biconcave discs population one could observe an increase of the number of transformed cells at different degeneration stages. Researches have shown that in mice with a pttg gene knockout compared with a control group of animals cytoskeletal protein--beta-spectrin was reduced by 17.03%. However, there is a reduction of membrane protein band 3 by 33.04%, simultaneously the content of anion transport protein band 4.5 increases by 35.2% and protein band 4.2 by 32.1%. The lectin blot analysis has helped to reveal changes in the structure of the carbohydrate determinants of erythrocyte membrane glycoproteins under conditions of directed pttg gene inactivation, accompanied by changes in the type of communication, which joins the terminal residue in carbohydrate determinant of glycoproteins. Thus, a significant redistribution of protein and fatty acids contents in erythrocyte membranes that manifested in the increase of the deformed shape of red blood cells is observed underpttg gene knockout.

Toward Effective Shell Modeling of Wrinkled Thin-Film Membranes Exhibiting Stress Concentrations

NASA Technical Reports Server (NTRS)

Tessler, Alexander; Sleight, David W.

2004-01-01

Geometrically nonlinear shell finite element analysis has recently been applied to solar-sail membrane problems in order to model the out-of-plane deformations due to structural wrinkling. Whereas certain problems lend themselves to achieving converged nonlinear solutions that compare favorably with experimental observations, solutions to tensioned membranes exhibiting high stress concentrations have been difficult to obtain even with the best nonlinear finite element codes and advanced shell element technology. In this paper, two numerical studies are presented that pave the way to improving the modeling of this class of nonlinear problems. The studies address the issues of mesh refinement and stress-concentration alleviation, and the effects of these modeling strategies on the ability to attain converged nonlinear deformations due to wrinkling. The numerical studies demonstrate that excessive mesh refinement in the regions of stress concentration may be disadvantageous to achieving wrinkled equilibrium states, causing the nonlinear solution to lock in the membrane response mode, while totally discarding the very low-energy bending response that is necessary to cause wrinkling deformation patterns. An element-level, strain-energy density criterion is suggested for facilitating automated, adaptive mesh refinements specifically aimed at the modeling of thin-film membranes undergoing wrinkling deformations.

Autonomous buckling of micrometer-sized lipid-protein membrane patches constructed by Dictyostelium discoideum.

PubMed

Takahashi, Kei; Toyota, Taro

2015-01-01

The cytosol of amoeba cells controls the membrane deformation during their motion in vivo. To investigate such ability of the cytosol of amoeba cell, Dictyostelium discoideum (Dictyostelium), in vitro, we used lipids extracted from Dictyostelium and commercially available phospholipids, and prepared substrate-supported lipid membrane patches on the micrometer scale by spin coating. We found that the spin coater holder, which has pores (pore size = 3.1 mm) of negative pressure to hold the cover glass induced the concave surface of the cover glass. The membrane lipid patches were formed at each position in the vicinity of the holder pores and their sizes were in the range of 2.7 to 3.2 × 10(4) μm(2). After addition of the cytosol extracted from Dictyostelium to the lipid membrane patches, through time-lapse observation with a confocal laser scanning fluorescence microscope, we observed an autonomous buckling of the Dictyostelium lipid patches and localized behaviours of proteins found within. The current method serves as the novel technique for the preparation of film patches in which the positions of patches are controlled by the holder pores without fabricating, modifying, and arranging the chemical properties of the solution components of lipids. The findings imply that lipid-binding proteins in the cytosol were adsorbed and accumulated within the Dictyostelium lipid patches, inducing the transformation of the cell-sized patch.

Two interdependent mechanisms of antimicrobial activity allow for efficient killing in nylon-3-based polymeric mimics of innate immunity peptides ☆

PubMed Central

Lee, Michelle W.; Chakraborty, Saswata; Schmidt, Nathan W.; Murgai, Rajan; Gellman, Samuel H.; Wong, Gerard C.L.

2015-01-01

Novel synthetic mimics of antimicrobial peptides have been developed to exhibit structural properties and antimicrobial activity similar to those of natural antimicrobial peptides (AMPs) of the innate immune system. These molecules have a number of potential advantages over conventional antibiotics, including reduced bacterial resistance, cost-effective preparation, and customizable designs. In this study, we investigate a family of nylon-3 polymer-based antimicrobials. By combining vesicle dye leakage, bacterial permeation, and bactericidal assays with small-angle X-ray scattering (SAXS), we find that these polymers are capable of two interdependent mechanisms of action: permeation of bacterial membranes and binding to intracellular targets such as DNA, with the latter necessarily dependent on the former. We systemically examine polymer-induced membrane deformation modes across a range of lipid compositions that mimic both bacteria and mammalian cell membranes. The results show that the polymers' ability to generate negative Gaussian curvature (NGC), a topological requirement for membrane permeation and cellular entry, in model Escherichia coli membranes correlates with their ability to permeate membranes without complete membrane disruption and kill E. coli cells. Our findings suggest that these polymers operate with a concentration dependent mechanism of action: at low concentrations permeation and DNA binding occur without membrane disruption, while at high concentrations complete disruption of the membrane occurs. This article is part of a Special Issue entitled: Interfacially Active Peptides and Proteins. PMID:24743021

Formation and organization of protein domains in the immunological synapse

NASA Astrophysics Data System (ADS)

Carlson, Andreas; Mahadevan, L.

2014-11-01

The cellular basis for the adaptive immune response during antigen recognition relies on a specialized protein interface known as the immunological synapse. Here, we propose a minimal mathematical model for the dynamics of the IS that encompass membrane mechanics, hydrodynamics and protein kinetics. Simple scaling laws describe the dynamics of protein clusters as a function of membrane stiffness, rigidity of the adhesive proteins, and fluid flow in the synaptic cleft. Numerical simulations complement the scaling laws by quantifying the nucleation, growth and stabilization of proteins domains on the size of the cell. Direct comparison with experiment suggests that passive dynamics suffices to describe the short-time formation and organization of protein clusters, while the stabilization and long time dynamics of the synapse is likely determined by active cytoskeleton processes triggered by receptor binding. Our study reveals that the fluid flow generated by the interplay between membrane deformation and protein binding kinetics can assist immune cells in regulating protein sorting.

Comparative ultrastructural characterization of African horse sickness virus-infected mammalian and insect cells reveals a novel potential virus release mechanism from insect cells.

PubMed

Venter, E; van der Merwe, C F; Buys, A V; Huismans, H; van Staden, V

2014-03-01

African horse sickness virus (AHSV) is an arbovirus capable of successfully replicating in both its mammalian host and insect vector. Where mammalian cells show a severe cytopathic effect (CPE) following AHSV infection, insect cells display no CPE. These differences in cell death could be linked to the method of viral release, i.e. lytic or non-lytic, that predominates in a specific cell type. Active release of AHSV, or any related orbivirus, has, however, not yet been documented from insect cells. We applied an integrated microscopy approach to compare the nanomechanical and morphological response of mammalian and insect cells to AHSV infection. Atomic force microscopy revealed plasma membrane destabilization, integrity loss and structural deformation of the entire surface of infected mammalian cells. Infected insect cells, in contrast, showed no morphological differences from mock-infected cells other than an increased incidence of circular cavities present on the cell surface. Transmission electron microscopy imaging identified a novel large vesicle-like compartment within infected insect cells, not present in mammalian cells, containing viral proteins and virus particles. Extracellular clusters of aggregated virus particles were visualized adjacent to infected insect cells with intact plasma membranes. We propose that foreign material is accumulated within these vesicles and that their subsequent fusion with the cell membrane releases entrapped viruses, thereby facilitating a non-lytic virus release mechanism different from the budding previously observed in mammalian cells. This insect cell-specific defence mechanism contributes to the lack of cell damage observed in AHSV-infected insect cells.

Theoretical estimates of mechanical properties of the endothelial cell cytoskeleton.

PubMed Central

Satcher, R L; Dewey, C F

1996-01-01

Current modeling of endothelial cell mechanics does not account for the network of F-actin that permeates the cytoplasm. This network, the distributed cytoplasmic structural actin (DCSA), extends from apical to basal membranes, with frequent attachments. Stress fibers are intercalated within the network, with similar frequent attachments. The microscopic structure of the DCSA resembles a foam, so that the mechanical properties can be estimated with analogy to these well-studied systems. The moduli of shear and elastic deformations are estimated to be on the order of 10(5) dynes/cm2. This prediction agrees with experimental measurements of the properties of cytoplasm and endothelial cells reported elsewhere. Stress fibers can potentially increase the modulus by a factor of 2-10, depending on whether they act in series or parallel to the network in transmitting surface forces. The deformations produced by physiological flow fields are of insufficient magnitude to disrupt cell-to-cell or DCSA cross-linkages. The questions raised by this paradox, and the ramifications of implicating the previously unreported DCSA as the primary force transmission element are discussed. Images FIGURE 2 PMID:8804594

Mechanical and optical behavior of a tunable liquid lens using a variable cross section membrane: modeling results

NASA Astrophysics Data System (ADS)

Flores-Bustamante, Mario C.; Rosete-Aguilar, Martha; Calixto, Sergio

2016-03-01

A lens containing a liquid medium and having at least one elastic membrane as one of its components is known as an elastic membrane lens (EML). The elastic membrane may have a constant or variable thickness. The optical properties of the EML change by modifying the profile of its elastic membrane(s). The EML formed of elastic constant thickness membrane(s) have been studied extensively. However, EML information using elastic membrane of variable thickness is limited. In this work, we present simulation results of the mechanical and optical behavior of two EML with variable thickness membranes (convex-plane membranes). The profile of its surfaces were modified by liquid medium volume increases. The model of the convex-plane membranes, as well as the simulation of its mechanical behavior, were performed using Solidworks® software; and surface's points of the deformed elastic lens were obtained. Experimental stress-strain data, obtained from a silicone rubber simple tensile test, according to ASTM D638 norm, were used in the simulation. Algebraic expressions, (Schwarzschild formula, up to four deformation coefficients, in a cylindrical coordinate system (r, z)), of the meridional profiles of the first and second surfaces of the deformed convex-plane membranes, were obtained using the results from Solidworks® and a program in the software Mathematica®. The optical performance of the EML was obtained by simulation using the software OSLO® and the algebraic expressions obtained in Mathematica®.

Hydrophilic Channel Alignment of Perfluoronated Sulfonic-Acid Ionomers for Vanadium Redox Flow Batteries.

PubMed

So, Soonyong; Cha, Min Suc; Jo, Sang-Woo; Kim, Tae-Ho; Lee, Jang Yong; Hong, Young Taik

2018-06-13

It is known that uniaxially drawn perfluoronated sulfonic-acid ionomers (PFSAs) show diffusion anisotropy because of the aligned water channels along the deformation direction. We apply the uniaxially stretched membranes to vanadium redox flow batteries (VRFBs) to suppress the permeation of active species, vanadium ions through the transverse directions. The aligned water channels render much lower vanadium permeability, resulting in higher Coulombic efficiency (>98%) and longer self-discharge time (>250 h). Similar to vanadium ions, proton conduction through the membranes also decreases as the stretching ratio increases, but the thinned membranes show the enhanced voltage and energy efficiencies over the range of current density, 50-100 mA/cm 2 . Hydrophilic channel alignment of PFSAs is also beneficial for long-term cycling of VRFBs in terms of capacity retention and cell performances. This simple pretreatment of membranes offers an effective and facile way to overcome high vanadium permeability of PFSAs for VRFBs.

Periodontal Regeneration Using Periodontal Ligament Stem Cell-Transferred Amnion

PubMed Central

Iwasaki, Kengo; Yokoyama, Naoki; Tanaka, Yuichi; Taki, Atsuko; Honda, Izumi; Kimura, Yasuyuki; Takeda, Masaki; Akazawa, Keiko; Oda, Shigeru; Izumi, Yuichi; Morita, Ikuo

2014-01-01

Periodontal disease is characterized by the destruction of tooth supporting tissues. Regeneration of periodontal tissues using ex vivo expanded cells has been introduced and studied, although appropriate methodology has not yet been established. We developed a novel cell transplant method for periodontal regeneration using periodontal ligament stem cell (PDLSC)-transferred amniotic membrane (PDLSC-amnion). The aim of this study was to investigate the regenerative potential of PDLSC-amnion in a rat periodontal defect model. Cultured PDLSCs were transferred onto amniotic membranes using a glass substrate treated with polyethylene glycol and photolithography. The properties of PDLSCs were investigated by flow cytometry and in vitro differentiation. PDLSC-amnion was transplanted into surgically created periodontal defects in rat maxillary molars. Periodontal regeneration was evaluated by microcomputed tomography (micro-CT) and histological analysis. PDLSCs showed mesenchymal stem cell-like characteristics such as cell surface marker expression (CD90, CD44, CD73, CD105, CD146, and STRO-1) and trilineage differentiation ability (i.e., into osteoblasts, adipocytes, and chondrocytes). PDLSC-amnion exhibited a single layer of PDLSCs on the amniotic membrane and stability of the sheet even with movement and deformation caused by surgical instruments. We observed that the PDLSC-amnion enhanced periodontal tissue regeneration as determined by micro-CT and histology by 4 weeks after transplantation. These data suggest that PDLSC-amnion has therapeutic potential as a novel cell-based regenerative periodontal therapy. PMID:24032400

Periodontal regeneration using periodontal ligament stem cell-transferred amnion.

PubMed

Iwasaki, Kengo; Komaki, Motohiro; Yokoyama, Naoki; Tanaka, Yuichi; Taki, Atsuko; Honda, Izumi; Kimura, Yasuyuki; Takeda, Masaki; Akazawa, Keiko; Oda, Shigeru; Izumi, Yuichi; Morita, Ikuo

2014-02-01

Periodontal disease is characterized by the destruction of tooth supporting tissues. Regeneration of periodontal tissues using ex vivo expanded cells has been introduced and studied, although appropriate methodology has not yet been established. We developed a novel cell transplant method for periodontal regeneration using periodontal ligament stem cell (PDLSC)-transferred amniotic membrane (PDLSC-amnion). The aim of this study was to investigate the regenerative potential of PDLSC-amnion in a rat periodontal defect model. Cultured PDLSCs were transferred onto amniotic membranes using a glass substrate treated with polyethylene glycol and photolithography. The properties of PDLSCs were investigated by flow cytometry and in vitro differentiation. PDLSC-amnion was transplanted into surgically created periodontal defects in rat maxillary molars. Periodontal regeneration was evaluated by microcomputed tomography (micro-CT) and histological analysis. PDLSCs showed mesenchymal stem cell-like characteristics such as cell surface marker expression (CD90, CD44, CD73, CD105, CD146, and STRO-1) and trilineage differentiation ability (i.e., into osteoblasts, adipocytes, and chondrocytes). PDLSC-amnion exhibited a single layer of PDLSCs on the amniotic membrane and stability of the sheet even with movement and deformation caused by surgical instruments. We observed that the PDLSC-amnion enhanced periodontal tissue regeneration as determined by micro-CT and histology by 4 weeks after transplantation. These data suggest that PDLSC-amnion has therapeutic potential as a novel cell-based regenerative periodontal therapy.

A Piezoelectric Unimorph Deformable Mirror Concept by Wafer Transfer for Ultra Large Space Telescopes

NASA Technical Reports Server (NTRS)

Yang, Eui-Hyeok; Shcheglov, Kirill

2002-01-01

Future concepts of ultra large space telescopes include segmented silicon mirrors and inflatable polymer mirrors. Primary mirrors for these systems cannot meet optical surface figure requirements and are likely to generate over several microns of wavefront errors. In order to correct for these large wavefront errors, high stroke optical quality deformable mirrors are required. JPL has recently developed a new technology for transferring an entire wafer-level mirror membrane from one substrate to another. A thin membrane, 100 mm in diameter, has been successfully transferred without using adhesives or polymers. The measured peak-to-valley surface error of a transferred and patterned membrane (1 mm x 1 mm x 0.016 mm) is only 9 nm. The mirror element actuation principle is based on a piezoelectric unimorph. A voltage applied to the piezoelectric layer induces stress in the longitudinal direction causing the film to deform and pull on the mirror connected to it. The advantage of this approach is that the small longitudinal strains obtainable from a piezoelectric material at modest voltages are thus translated into large vertical displacements. Modeling is performed for a unimorph membrane consisting of clamped rectangular membrane with a PZT layer with variable dimensions. The membrane transfer technology is combined with the piezoelectric bimorph actuator concept to constitute a compact deformable mirror device with a large stroke actuation of a continuous mirror membrane, resulting in a compact A0 systems for use in ultra large space telescopes.

Modelling cell motility and chemotaxis with evolving surface finite elements

PubMed Central

Elliott, Charles M.; Stinner, Björn; Venkataraman, Chandrasekhar

2012-01-01

We present a mathematical and a computational framework for the modelling of cell motility. The cell membrane is represented by an evolving surface, with the movement of the cell determined by the interaction of various forces that act normal to the surface. We consider external forces such as those that may arise owing to inhomogeneities in the medium and a pressure that constrains the enclosed volume, as well as internal forces that arise from the reaction of the cells' surface to stretching and bending. We also consider a protrusive force associated with a reaction–diffusion system (RDS) posed on the cell membrane, with cell polarization modelled by this surface RDS. The computational method is based on an evolving surface finite-element method. The general method can account for the large deformations that arise in cell motility and allows the simulation of cell migration in three dimensions. We illustrate applications of the proposed modelling framework and numerical method by reporting on numerical simulations of a model for eukaryotic chemotaxis and a model for the persistent movement of keratocytes in two and three space dimensions. Movies of the simulated cells can be obtained from http://homepages.warwick.ac.uk/∼maskae/CV_Warwick/Chemotaxis.html. PMID:22675164

Tunable-focus liquid lens controlled using a servo motor

NASA Astrophysics Data System (ADS)

Ren, Hongwen; Fox, David; Anderson, P. Andrew; Wu, Benjamin; Wu, Shin-Tson

2006-09-01

We demonstrated a liquid lens whose focal length can be controlled by an actuator. The lens cell is composed of elastic membrane, planar glass plate, a periphery sealing ring, and a liquid with a fixed volume in the lens chamber. Part of the periphery sealing ring is excavated to form a hollow chamber which functions as a reservoir. This hollowed periphery is surrounded by an exterior rubber membrane. The shaft of an actuator is used to deform the elastic rubber. Squeezing the liquid contained in the reservoir into the lens chamber. Excess liquid in the lens chamber will push the lens membrane to outward, resulting in a lens shape change. Due to the compact structure and easy operation, this liquid lens has potential applications in zoom lenses, auto beam steering, and eyeglasses.

Hemoglobin redox reactions and red blood cell aging.

PubMed

Rifkind, Joseph M; Nagababu, Enika

2013-06-10

The physiological mechanism(s) for recognition and removal of red blood cells (RBCs) from circulation after 120 days of its lifespan is not fully understood. Many of the processes thought to be associated with the removal of RBCs involve oxidative stress. We have focused on hemoglobin (Hb) redox reactions, which is the major source of RBC oxidative stress. The importance of Hb redox reactions have been shown to originate in large parts from the continuous slow autoxidation of Hb producing superoxide and its dramatic increase under hypoxic conditions. In addition, oxidative stress has been shown to be associated with redox reactions that originate from Hb reactions with nitrite and nitric oxide (NO) and the resultant formation of highly toxic peroxynitrite when NO reacts with superoxide released during Hb autoxidation. The interaction of Hb, particularly under hypoxic conditions with band 3 of the RBC membrane is critical for the generating the RBC membrane changes that trigger the removal of cells from circulation. These changes include exposure of antigenic sites, increased calcium leakage into the RBC, and the resultant leakage of potassium out of the RBC causing cell shrinkage and impaired deformability. The need to understand the oxidative damage to specific membrane proteins that result from redox reactions occurring when Hb is bound to the membrane. Proteomic studies that can pinpoint the specific proteins damaged under different conditions will help elucidate the cellular aging processes that result in cells being removed from circulation.

Multiscale Modeling of Primary Cilium Deformations Under Local Forces and Shear Flows

NASA Astrophysics Data System (ADS)

Peng, Zhangli; Feng, Zhe; Resnick, Andrew; Young, Yuan-Nan

2017-11-01

We study the detailed deformations of a primary cilium under local forces and shear flows by developing a multiscale model based on the state-of-the-art understanding of its molecular structure. Most eukaryotic cells are ciliated with primary cilia. Primary cilia play important roles in chemosensation, thermosensation, and mechanosensation, but the detailed mechanism for mechanosensation is not well understood. We apply the dissipative particle dynamics (DPD) to model an entire well with a primary cilium and consider its different components, including the basal body, microtubule doublets, actin cortex, and lipid bilayer. We calibrate the mechanical properties of individual components and their interactions from experimental measurements and molecular dynamics simulations. We validate the simulations by comparing the deformation profile of the cilium and the rotation of the basal body with optical trapping experiments. After validations, we investigate the deformation of the primary cilium under shear flows. Furthermore, we calculate the membrane tensions and cytoskeleton stresses, and use them to predict the activation of mechanosensitive channels.

TGF-β induces changes in breast cancer cell deformability.

PubMed

Kulkarni, Ankur; Chatterjee, Aritra; Kondaiah, Paturu; Gundiah, Namrata

2018-05-10

Mechanical properties of cells regulate cell behaviors which lead to phenotypic changes that may aid in the development and progression of disease. In this study, we used atomic force microscopy (AFM) indentation with a spherical probe to characterize the elastic and viscoelastic properties of invasive (MDA-MB-231) and noninvasive (MCF-7) breast cancer cells treated with transforming growth factor-β (TGF-β). We also used confocal fluorescence imaging to investigate the sub-membrane cytoskeletal structure of the cells. Results showed significant alterations in moduli of both cell types after 24 hour TGF-β treatment which had a context dependent response; moduli for MDA-MB-231 decreased whereas MCF-7 demonstrated stiffening response. Viscoelastic characterization using stress relaxation tests showed increased fluid-like nature of MDA-MB-231 following TGF-β treatment and lower fluidity for MCF-7 cells. We also observed significant alterations in the expression and orientation of actin stress fibers with TGF-β treatment which correlated with the changes in cell mechanics. The less invasive MCF-7 cells had a delayed overall increase in cell deformability after 48 hour exposure to TGF-β; a similar trend was observed for MDA-MB cells. These changes may be important to facilitate migration, for instance, during metastasis of cancer cells through submicron sized spaces. © 2018 IOP Publishing Ltd.

Large-deformation electrohydrodynamics of an elastic capsule in a DC electric field

NASA Astrophysics Data System (ADS)

Das, Sudip; Thaokar, Rochish M.

2018-04-01

The dynamics of a spherical elastic capsule, containing a Newtonian fluid bounded by an elastic membrane and immersed in another Newtonian fluid, in a uniform DC electric field is investigated. Discontinuity of electrical properties such as conductivities of the internal and external fluid media as well as capacitance and conductance of the membrane lead to a net interfacial Maxwell stress which can cause the deformation of such an elastic capsule. We investigate this problem considering well established membrane laws for a thin elastic membrane, with fully resolved hydrodynamics in the Stokes flow limit and describe the electrostatics using the capacitor model. In the limit of small deformation, the analytical theory predicts the dynamics fairly satisfactorily. Large deformations at high capillary number though necessitate a numerical approach (Boundary element method in the present case) to solve this highly non-linear problem. Akin to vesicles, at intermediate times, highly nonlinear biconcave shapes along with squaring and hexagon like shapes are observed when the outer medium is more conducting. The study identifies the essentiality of parameters such as high membrane capacitance, low membrane conductance, low hydrodynamic time scales and high capillary number for observation of these shape transitions. The transition is due to large compressive Maxwell stress at the poles at intermediate times. Thus such shape transition can be seen in spherical globules admitting electrical capacitance, possibly, irrespective of the nature of the interfacial restoring force.

Preliminary Characterization of Erythrocytes Deformability on the Entropy-Complexity Plane

PubMed Central

Korol, Ana M; D’Arrigo, Mabel; Foresto, Patricia; Pérez, Susana; Martín, Maria T; Rosso, Osualdo A

2010-01-01

We present an application of wavelet-based Information Theory quantifiers (Normalized Total Shannon Entropy, MPR-Statistical Complexity and Entropy-Complexity plane) on red blood cells membrane viscoelasticity characterization. These quantifiers exhibit important localization advantages provided by the Wavelet Theory. The present approach produces a clear characterization of this dynamical system, finding out an evident manifestation of a random process on the red cell samples of healthy individuals, and its sharp reduction of randomness on analyzing a human haematological disease, such as β-thalassaemia minor. PMID:21611139

A novel design of membrane mirror with small deformation and imaging performance analysis in infrared system

NASA Astrophysics Data System (ADS)

Zhang, Shuqing; Wang, Yongquan; Zhi, Xiyang

2017-05-01

A method of diminishing the shape error of membrane mirror is proposed in this paper. The inner inflating pressure is considerably decreased by adopting the pre-shaped membrane. Small deformation of the membrane mirror with greatly reduced shape error is sequentially achieved. Primarily a finite element model of the above pre-shaped membrane is built on the basis of its mechanical properties. Then accurate shape data under different pressures can be acquired by iteratively calculating the node displacements of the model. Shape data are applicable to build up deformed reflecting surfaces for the simulative analysis in ZEMAX. Finally, ground-based imaging experiments of 4-bar targets and nature scene are conducted. Experiment results indicate that the MTF of the infrared system can reach to 0.3 at a high spatial resolution of 10l p/mm, and texture details of the nature scene are well-presented. The method can provide theoretical basis and technical support for the applications in lightweight optical components with ultra-large apertures.

The bio-response of osteocytes and its regulation on osteoblasts under vibration.

PubMed

Wu, Xin-Tong; Sun, Lian-Wen; Qi, Hong-Yu; Shi, Hao; Fan, Yu-Bo

2016-04-01

Vibration, especially at low magnitude and high frequency (LMHF), was demonstrated to be anabolic for bone, but how the LMHF vibration signal is perceived by osteocytes is not fully studied. On the other hand, the mechanotransduction of osteocytes under shear stress has been scientists' primary focus for years. Due to the small strain caused by low-magnitude vibration, whether the previous explanation for shear stress will still work for LMHF vibration is unknown. In this study, a finite element method (FEM) model based on the real geometrical shape of an osteocyte was built to compare the mechanical behaviors of osteocytes under LMHF vibration and shear stress. The bio-response of osteocytes to vibration under different frequencies, including the secretion of soluble factors and the concentration of intracellular calcium, were studied. The regulating effect of the conditioned medium (CM) from vibrated osteocytes on osteoblasts was also studied. The FEM analysis result showed the cell membrane deformation under LMHF vibration was very small (with a peak value of 1.09%) as compared to the deformation caused by shear stress (with a peak value of 6.65%). The F-actin stress fibers of osteocytes were reorganized, especially on the nucleus periphery after LMHF vibration. The vibration at 30 Hz has a promoting effect on osteocytes and the osteogenesis of osteoblasts, whereas vibration at 90 Hz was suppressive. These results lead to a conclusion that the bio-response of osteocytes to LMHF vibration is frequency-dependent and is more related to the cytoskeleton on nuclear periphery rather than the membrane deformation. © 2016 International Federation for Cell Biology.

Discrete Element Framework for Modelling Extracellular Matrix, Deformable Cells and Subcellular Components

PubMed Central

Gardiner, Bruce S.; Wong, Kelvin K. L.; Joldes, Grand R.; Rich, Addison J.; Tan, Chin Wee; Burgess, Antony W.; Smith, David W.

2015-01-01

This paper presents a framework for modelling biological tissues based on discrete particles. Cell components (e.g. cell membranes, cell cytoskeleton, cell nucleus) and extracellular matrix (e.g. collagen) are represented using collections of particles. Simple particle to particle interaction laws are used to simulate and control complex physical interaction types (e.g. cell-cell adhesion via cadherins, integrin basement membrane attachment, cytoskeletal mechanical properties). Particles may be given the capacity to change their properties and behaviours in response to changes in the cellular microenvironment (e.g., in response to cell-cell signalling or mechanical loadings). Each particle is in effect an ‘agent’, meaning that the agent can sense local environmental information and respond according to pre-determined or stochastic events. The behaviour of the proposed framework is exemplified through several biological problems of ongoing interest. These examples illustrate how the modelling framework allows enormous flexibility for representing the mechanical behaviour of different tissues, and we argue this is a more intuitive approach than perhaps offered by traditional continuum methods. Because of this flexibility, we believe the discrete modelling framework provides an avenue for biologists and bioengineers to explore the behaviour of tissue systems in a computational laboratory. PMID:26452000

Discrete Element Framework for Modelling Extracellular Matrix, Deformable Cells and Subcellular Components.

PubMed

Gardiner, Bruce S; Wong, Kelvin K L; Joldes, Grand R; Rich, Addison J; Tan, Chin Wee; Burgess, Antony W; Smith, David W

2015-10-01

This paper presents a framework for modelling biological tissues based on discrete particles. Cell components (e.g. cell membranes, cell cytoskeleton, cell nucleus) and extracellular matrix (e.g. collagen) are represented using collections of particles. Simple particle to particle interaction laws are used to simulate and control complex physical interaction types (e.g. cell-cell adhesion via cadherins, integrin basement membrane attachment, cytoskeletal mechanical properties). Particles may be given the capacity to change their properties and behaviours in response to changes in the cellular microenvironment (e.g., in response to cell-cell signalling or mechanical loadings). Each particle is in effect an 'agent', meaning that the agent can sense local environmental information and respond according to pre-determined or stochastic events. The behaviour of the proposed framework is exemplified through several biological problems of ongoing interest. These examples illustrate how the modelling framework allows enormous flexibility for representing the mechanical behaviour of different tissues, and we argue this is a more intuitive approach than perhaps offered by traditional continuum methods. Because of this flexibility, we believe the discrete modelling framework provides an avenue for biologists and bioengineers to explore the behaviour of tissue systems in a computational laboratory.

Structural and experimental investigations of the functional anatomy and the turgor of the notochord in the larval tail of anuran tadpoles.

PubMed

Platz, Franz

2006-07-01

The premetamorphotic morphology and metamorphotic degeneration of the tail notochord of anuran tadpoles has been investigated. For this purpose the functional anatomy and origin of the notochord turgor was analysed in 10 species macroscopically and using light, transmission and scanning electron microscopic techniques. The notochord consists of the fibrous notochord sheath, which surrounds the notochord cells. Within the sheath these cells form a net-like unit. The inner cells are derived from the marginal notochord cells (chordoblasts). They are protected from mechanical overload by intracellular filaments and desmosomes. Due to their vacuoles, which are filled with a hyaline liquid, they have a constant volume but are deformable. Dissolved substances may pass from the vascularized fin to the notochord cells. The transport from marginal to inner cells occurs via cytopempsis and micropinocytosis. The morphological correlation of this process consists of multiple membrane invaginations and intracellular vesicles. Within the notochord cells a high turgor pressure has been observed. During metamorphosis the membrane vesiculation persists and the notochord cells degenerate. Due to the loss of turgor pressure the tight consistency of the notochord is lost. The collagen filaments and the elastic membrane of the notochord sheath dissolve. Notochord cells with their filaments, high turgor pressure and their central vacuole can function as a combined mechanical and physiological system, which is adaptable to the needs of pressure, compression, tensile and bending forces.

MEMS deformable mirror for wavefront correction of large telescopes

NASA Astrophysics Data System (ADS)

Manhart, Sigmund; Vdovin, Gleb; Collings, Neil; Sodnik, Zoran; Nikolov, Susanne; Hupfer, Werner

2017-11-01

A 50 mm diameter membrane mirror was designed and manufactured at TU Delft. It is made from bulk silicon by micromachining - a technology primarily used for micro-electromechanical systems (MEMS). The mirror unit is equipped with 39 actuator electrodes and can be electrostatically deformed to correct wavefront errors in optical imaging systems. Performance tests on the deformable mirror were carried out at Astrium GmbH using a breadboard setup with a wavefront sensor and a closed-loop control system. It was found that the deformable membrane mirror is well suited for correction of low order wavefront errors as they must be expected in lightweighted space telescopes.

Observing polymersome dynamics in controlled microscale flows

NASA Astrophysics Data System (ADS)

Kumar, Subhalakshmi; Shenoy, Anish; Schroeder, Charles

2015-03-01

Achieving an understanding of single particle rheology for large yet deformable particles with controlled membrane viscoelasticity is major challenge in soft materials. In this work, we directly visualize the dynamics of single polymersomes (~ 10 μm in size) in an extensional flow using optical microscopy. We generate polymer vesicular structures composed of polybutadiene-block-polyethylene oxide (PB-b-PEO) copolymers. Single polymersomes are confined near the stagnation point of a planar extensional flow using an automated microfluidic trap, thereby enabling the direct observation of polymersome dynamics under fluid flows with controlled strains and strain rates. In a series of experiments, we investigate the effect of varying elasticity in vesicular membranes on polymersome deformation, along with the impact of decreasing membrane fluidity upon increasing diblock copolymer molecular weight. Overall, we believe that this approach will enable precise characterization of the role of membrane properties on single particle rheology for deformable polymersomes.

Microscopic mechanism analyses on influence of metabolism of erythrocyte membrane-lipid etc. by LLLIB

NASA Astrophysics Data System (ADS)

Xu, Lin; Zhang, Canbang; Wen, Yuanbin; Liu, Shuxiao; Zhou, Lingyun

2009-08-01

Some cases with cerebral infarction were treated by He-Ne laser irradiation on blood. In the treatment before and after, membrane-cholesterol(C)/membrane-phosphatide(P), membrane fluidity(F) and deformability of erythrocyte were determined. The results showed that low level laser irradiation on blood (LLLIB) can sure reduce the ratio of (C)/(P), can heighten fluidity and improve deformability of erythrocyte .Thus the metabolism ability of erythrocyte membrane-lipid ,the blood circulation and the properties of hemorheology can be improved. In this paper, the microscopic mechanism of those aforesaid action effects by low level laser irradiation on blood were analyzed by means of Quantum theory and some corresponding models.

Molecular dynamics simulations showing 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) membrane mechanoporation damage under different strain paths.

PubMed

Murphy, M A; Mun, Sungkwang; Horstemeyer, M F; Baskes, M I; Bakhtiary, A; LaPlaca, Michelle C; Gwaltney, Steven R; Williams, Lakiesha N; Prabhu, R K

2018-04-09

Continuum finite element material models used for traumatic brain injury lack local injury parameters necessitating nanoscale mechanical injury mechanisms be incorporated. One such mechanism is membrane mechanoporation, which can occur during physical insults and can be devastating to cells, depending on the level of disruption. The current study investigates the strain state dependence of phospholipid bilayer mechanoporation and failure. Using molecular dynamics, a simplified membrane, consisting of 72 1-palmitoyl-2-oleoyl-phosphatidylcholine (POPC) phospholipids, was subjected to equibiaxial, 2:1 non-equibiaxial, 4:1 non-equibiaxial, strip biaxial, and uniaxial tensile deformations at a von Mises strain rate of 5.45 × 10 8 s -1 , resulting in velocities in the range of 1 to 4.6 m·s -1 . A water bridge forming through both phospholipid bilayer leaflets was used to determine structural failure. The stress magnitude, failure strain, headgroup clustering, and damage responses were found to be strain state-dependent. The strain state order of detrimentality in descending order was equibiaxial, 2:1 non-equibiaxial, 4:1 non-equibiaxial, strip biaxial, and uniaxial. The phospholipid bilayer failed at von Mises strains of .46, .47, .53, .77, and 1.67 during these respective strain path simulations. Additionally, a Membrane Failure Limit Diagram (MFLD) was created using the pore nucleation, growth, and failure strains to demonstrate safe and unsafe membrane deformation regions. This MFLD allowed representative equations to be derived to predict membrane failure from in-plane strains. These results provide the basis to implement a more accurate mechano-physiological internal state variable continuum model that captures lower length scale damage and will aid in developing higher fidelity injury models.

Pseudomonas aeruginosa inactivation mechanism is affected by capsular extracellular polymeric substances reactivity with chlorine and monochloramine.

PubMed

Xue, Zheng; Hessler, Christopher M; Panmanee, Warunya; Hassett, Daniel J; Seo, Youngwoo

2013-01-01

The reactivity of capsular extracellular polymeric substances (EPS) to chlorine and monochloramine was assessed and compared in this study. The impact of capsular EPS on Gram-negative bacteria Pseudomonas aeruginosa inactivation mechanisms was investigated both qualitatively and quantitatively using a combination of batch experiments, viability tests with LIVE/DEAD staining, and Fourier transform infrared spectroscopy (FTIR). Both wild-type and isogenic mutant strains with different alginate EPS production capabilities were used to evaluate their susceptibility to chlorine and monochloramine. The mucA22 mutant strain, which overproduces the EPS composed largely of acidic polysaccharide alginate, exhibited high resistance and prolonged inactivation time to both chlorine and monochloramine relative to PAO1 (wild-type) and algT(U) mutant strains (alginate EPS deficient). Multiple analyses were combined to better understand the mechanistic role of EPS against chlorine-based disinfectants. The extracted EPS exhibited high reactivity with chlorine and very low reactivity with monochloramine, suggesting different mechanism of protection against disinfectants. Moreover, capsular EPS on cell membrane appeared to reduce membrane permeabilization by disinfectants as suggested by deformation of key functional groups in EPS and cell membrane (the C-O-C stretching of carbohydrate and the C=O stretching of ester group). The combined results supported that capsular EPS, acting either as a disinfectant consumer (for chlorine inactivation) or limiting access to reactive sites on cell membrane (for monochloramine inactivation), provide a protective role for bacterial cells against regulatory residual disinfectants by reducing membrane permeabilization. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Generalized Boussinesq-Scriven surface fluid model with curvature dissipation for liquid surfaces and membranes.

PubMed

Aguilar Gutierrez, Oscar F; Herrera Valencia, Edtson E; Rey, Alejandro D

2017-10-01

Curvature dissipation is relevant in synthetic and biological processes, from fluctuations in semi-flexible polymer solutions, to buckling of liquid columns, tomembrane cell wall functioning. We present a micromechanical model of curvature dissipation relevant to fluid membranes and liquid surfaces based on a parallel surface parameterization and a stress constitutive equation appropriate for anisotropic fluids and fluid membranes.The derived model, aimed at high curvature and high rate of change of curvature in liquid surfaces and membranes, introduces additional viscous modes not included in the widely used 2D Boussinesq-Scriven rheological constitutive equation for surface fluids.The kinematic tensors that emerge from theparallel surface parameterization are the interfacial rate of deformation and the surface co-rotational Zaremba-Jaumann derivative of the curvature, which are used to classify all possibledissipative planar and non-planar modes. The curvature dissipation function that accounts for bending, torsion and twist rates is derived and analyzed under several constraints, including the important inextensional bending mode.A representative application of the curvature dissipation model to the periodic oscillation in nano-wrinkled outer hair cells show how and why curvature dissipation decreases with frequency, and why the 100kHz frequency range is selected. These results contribute to characterize curvature dissipation in membranes and liquid surfaces. Copyright © 2017 Elsevier Inc. All rights reserved.

Membrane Crossing and Membranotropic Activity of Cell-Penetrating Peptides: Dangerous Liaisons?

PubMed

Walrant, Astrid; Cardon, Sébastien; Burlina, Fabienne; Sagan, Sandrine

2017-12-19

Living organisms have to maintain a stable balance in molecules and ions in the changing environment in which they are living, a process known as homeostasis. At the level of cells, the plasma membrane has a major role in homeostasis, since this hydrophobic film prevents passive diffusion of large and hydrophilic molecules between the extracellular and intracellular milieu. Living organisms have evolved with highly sophisticated transport systems to control exchanges across this barrier: import of nutrients and fuel essential for their survival; recognition of chemical or physical messengers allowing information interchanges with surrounding cells. Besides specialized proteins, endocytosis mechanisms at the level of the lipid bilayer can transport molecules from the outside across the cell membrane, in an energy-dependent manner. The cell membrane is highly heterogeneous in its molecular composition (tens of different lipids, proteins, polysaccharides, and combinations of these) and dynamic with bending, deformation, and elastic properties that depend on the local composition of membrane domains. Many viruses, microorganisms, and toxins exploit the plasma membrane to enter into cells. Chemists develop strategies to target the plasma membrane with molecules capable of circumventing this hydrophobic barrier, in particular to transport and deliver nonpermeable drugs in cells for biotechnological or pharmaceutical purposes. Drug delivery systems are numerous and include lipid-, sugar-, protein-, and peptide-based delivery systems, since these biomolecules generally have good biocompatibility, biodegradability, environmental sustainability, cost effectiveness, and availability. Among those, cell-penetrating peptides (CPPs), reported for the first time in the early 1990s, are attracting major interest not only as potential drug delivery systems but also at the level of fundamental research. It was indeed demonstrated very early that these peptides, which generally correspond to highly cationic sequences, can still cross the cell membrane at 4 °C, a temperature at which all active transport and endocytosis pathways are totally inhibited. Therefore, how these charged hydrophilic peptides cross the hydrophobic membrane barrier is of utmost interest as a pure basic and physicochemical question. In this Account, we focus on cationic cell-penetrating peptides (CPPs) and the way they cross cell membranes. We summarize the history of this field that emerged around 20 years ago. CPPs were indeed first identified as protein-transduction domains from the human immunodeficiency virus (HIV) TAT protein and the Antennapedia homeoprotein, a transcription factor from Drosophila. We highlight our contribution to the elucidation of CPP internalization pathways, in particular translocation, which implies perturbation and reorganization of the lipid bilayer, and endocytosis depending on sulfated glycosaminoglycans. We show a particular role of Trp (indole side chain) and Arg (guanidinium side chain), which are essential amino acids for CPP internalization. Interactions with the cell-surface are not only Coulombic; H-bonds and hydrophobic interactions contribute also significantly to CPP entry. The capacity of CPPs to cross cell membrane is not related to their strength of membrane binding. Finally, we present optimized methods based on mass spectrometry and fluorescence spectroscopy that allow unequivocal quantification of CPPs inside cells or bound to the outer leaflet of the membrane, and discuss some limitations of the technique of flow cytometry that we have recently highlighted.

On the mechanochemical theory of biological pattern formation with application to vasculogenesis.

PubMed

Murray, James D

2003-02-01

We first describe the Murray-Oster mechanical theory of pattern formation, the biological basis of which is experimentally well documented. The model quantifies the interaction of cells and the extracellular matrix via the cell-generated forces. The model framework is described in quantitative detail. Vascular endothelial cells, when cultured on gelled basement membrane matrix, rapidly aggregate into clusters while deforming the matrix into a network of cord-like structures tessellating the planar culture. We apply the mechanical theory of pattern formation to this culture system and show that neither strain-biased anisotropic cell traction nor cell migration are necessary for pattern formation: isotropic, strain-stimulated cell traction is sufficient to form the observed patterns. Predictions from the model were confirmed experimentally.

Rheology of dilute suspensions of red blood cells: experimental and theoretical approaches

NASA Astrophysics Data System (ADS)

Drochon, A.

2003-05-01

Shear viscosity measurements with dilute suspensions of red blood cells are interpreted using a microrheological model that relates the bulk measurements to the physical properties of the suspended cells. It is thus possible to quantify the average deformability of a RBC population in terms of a mean value of the membrane shear elastic modulus E_s. The values obtained for normal cells are in good agreement with those given in the literature. The method allows to discriminate between normal and altered (diamide or glutaraldehyde treated) cells or pathological cells (scleroderma). The predictions of the microrheological model, based on analytic calculations, are also compared with the numerical results of Ramanujan and Pozrikidis (JFM 361, 1998) for dilute suspensions of capsules in simple shear flow.

Dynamic actin filaments control the mechanical behavior of the human red blood cell membrane

PubMed Central

Gokhin, David S.; Nowak, Roberta B.; Khoory, Joseph A.; de la Piedra, Alfonso; Ghiran, Ionita C.; Fowler, Velia M.

2015-01-01

Short, uniform-length actin filaments function as structural nodes in the spectrin-actin membrane skeleton to optimize the biomechanical properties of red blood cells (RBCs). Despite the widespread assumption that RBC actin filaments are not dynamic (i.e., do not exchange subunits with G-actin in the cytosol), this assumption has never been rigorously tested. Here we show that a subpopulation of human RBC actin filaments is indeed dynamic, based on rhodamine-actin incorporation into filaments in resealed ghosts and fluorescence recovery after photobleaching (FRAP) analysis of actin filament mobility in intact RBCs (∼25–30% of total filaments). Cytochalasin-D inhibition of barbed-end exchange reduces rhodamine-actin incorporation and partially attenuates FRAP recovery, indicating functional interaction between actin subunit turnover at the single-filament level and mobility at the membrane-skeleton level. Moreover, perturbation of RBC actin filament assembly/disassembly with latrunculin-A or jasplakinolide induces an approximately twofold increase or ∼60% decrease, respectively, in soluble actin, resulting in altered membrane deformability, as determined by alterations in RBC transit time in a microfluidic channel assay, as well as by abnormalities in spontaneous membrane oscillations (flickering). These experiments identify a heretofore-unrecognized but functionally important subpopulation of RBC actin filaments, whose properties and architecture directly control the biomechanical properties of the RBC membrane. PMID:25717184

Ethylhexylglycerin Impairs Membrane Integrity and Enhances the Lethal Effect of Phenoxyethanol

PubMed Central

Langsrud, Solveig; Steinhauer, Katrin; Lüthje, Sonja; Weber, Klaus; Goroncy-Bermes, Peter; Holck, Askild L.

2016-01-01

Preservatives are added to cosmetics to protect the consumers from infections and prevent product spoilage. The concentration of preservatives should be kept as low as possible and this can be achieved by adding potentiating agents. The aim of the study was to investigate the mechanisms behind potentiation of the bactericidal effect of a commonly used preservative, 2-phenoxyethanol (PE), by the potentiating agent ethylhexylglycerin (EHG). Sub-lethal concentrations of EHG (0.075%) and PE (0.675%) in combination led to rapid killing of E. coli (> 5 log reduction of cfu after 30 min), leakage of cellular constituents, disruption of the energy metabolism, morphological deformities of cells and condensation of DNA. Used alone, EHG disrupted the membrane integrity even at low concentrations. In conclusion, sub-lethal concentrations of EHG potentiate the effect of PE through damage of the cell membrane integrity. Thus, adding EHG to PE in a 1:9 ratio has a similar effect on membrane damage and bacterial viability as doubling the concentration of PE. This study provides insight about the mechanism of action of a strong potentiating agent, EHG, which is commonly used in cosmetics together with PE. PMID:27783695

Two interdependent mechanisms of antimicrobial activity allow for efficient killing in nylon-3-based polymeric mimics of innate immunity peptides.

PubMed

Lee, Michelle W; Chakraborty, Saswata; Schmidt, Nathan W; Murgai, Rajan; Gellman, Samuel H; Wong, Gerard C L

2014-09-01

Novel synthetic mimics of antimicrobial peptides have been developed to exhibit structural properties and antimicrobial activity similar to those of natural antimicrobial peptides (AMPs) of the innate immune system. These molecules have a number of potential advantages over conventional antibiotics, including reduced bacterial resistance, cost-effective preparation, and customizable designs. In this study, we investigate a family of nylon-3 polymer-based antimicrobials. By combining vesicle dye leakage, bacterial permeation, and bactericidal assays with small-angle X-ray scattering (SAXS), we find that these polymers are capable of two interdependent mechanisms of action: permeation of bacterial membranes and binding to intracellular targets such as DNA, with the latter necessarily dependent on the former. We systemically examine polymer-induced membrane deformation modes across a range of lipid compositions that mimic both bacteria and mammalian cell membranes. The results show that the polymers' ability to generate negative Gaussian curvature (NGC), a topological requirement for membrane permeation and cellular entry, in model Escherichia coli membranes correlates with their ability to permeate membranes without complete membrane disruption and kill E. coli cells. Our findings suggest that these polymers operate with a concentration-dependent mechanism of action: at low concentrations permeation and DNA binding occur without membrane disruption, while at high concentrations complete disruption of the membrane occurs. This article is part of a Special Issue entitled: Interfacially Active Peptides and Proteins. Guest Editors: William C. Wimley and Kalina Hristova. Copyright © 2014 Elsevier B.V. All rights reserved.

Mutations in BIN1 Associated with Centronuclear Myopathy Disrupt Membrane Remodeling by Affecting Protein Density and Oligomerization

PubMed Central

Wu, Tingting; Shi, Zheng; Baumgart, Tobias

2014-01-01

The regulation of membrane shapes is central to many cellular phenomena. Bin/Amphiphysin/Rvs (BAR) domain-containing proteins are key players for membrane remodeling during endocytosis, cell migration, and endosomal sorting. BIN1, which contains an N-BAR domain, is assumed to be essential for biogenesis of plasma membrane invaginations (T-tubules) in muscle tissues. Three mutations, K35N, D151N and R154Q, have been discovered so far in the BAR domain of BIN1 in patients with centronuclear myopathy (CNM), where impaired organization of T-tubules has been reported. However, molecular mechanisms behind this malfunction have remained elusive. None of the BIN1 disease mutants displayed a significantly compromised curvature sensing ability. However, two mutants showed impaired membrane tubulation both in vivo and in vitro, and displayed characteristically different behaviors. R154Q generated smaller membrane curvature compared to WT N-BAR. Quantification of protein density on membranes revealed a lower membrane-bound density for R154Q compared to WT and the other mutants, which appeared to be the primary reason for the observation of impaired deformation capacity. The D151N mutant was unable to tubulate liposomes under certain experimental conditions. At medium protein concentrations we found ‘budding’ structures on liposomes that we hypothesized to be intermediates during the tubulation process except for the D151N mutant. Chemical crosslinking assays suggested that the D151N mutation impaired protein oligomerization upon membrane binding. Although we found an insignificant difference between WT and K35N N-BAR in in vitro assays, depolymerizing actin in live cells allowed tubulation of plasma membranes through the K35N mutant. Our results provide insights into the membrane-involved pathophysiological mechanisms leading to human disease. PMID:24755653

Gratings Fabricated on Flat Surfaces and Reproduced on Non-Flat Substrates

NASA Technical Reports Server (NTRS)

Content, David; Iazikov, Dmitri; Mossberg, Thomas W.; Greiner, Christopher M.

2009-01-01

A method has been developed for fabricating gratings on flat substrates, and then reproducing the groove pattern on a curved (concave or convex) substrate and a corresponding grating device. First, surface relief diffraction grating grooves are formed on flat substrates. For example, they may be fabricated using photolithography and reactive ion etching, maskless lithography, holography, or mechanical ruling. Then, an imprint of the grating is made on a deformable substrate, such as plastic, polymer, or other materials using thermoforming, hot or cold embossing, or other methods. Interim stamps using electroforming, or other methods, may be produced for the imprinting process or if the same polarity of the grating image is required. The imprinted, deformable substrate is then attached to a curved, rigid substrate using epoxy or other suitable adhesives. The imprinted surface is facing away from the curved rigid substrate. As an alternative fabrication method, after grating is imprinted on the deformable substrate as described above, the grating may be coated with thin conformal conductive layer (for example, using vacuum deposition of gold). Then the membrane may be mounted over an opening in a pressured vessel in a manner of a membrane on a drum, grating side out. The pressure inside of the vessel may be changed with respect to the ambient pressure to produce concave or convex membrane surface. The shape of the opening may control the type of the surface curvature (for example, a circular opening would create spherical surface, oval opening would create toroidal surface, etc.). After that, well-known electroforming methods may be used to create a replica of the grating on the concave or convex membrane. For example, the pressure vessel assembly may be submerged into an electro-forming solution and negative electric potential applied to the metal coated membrane using an insulated wire. Positive electric potential may be then applied to a nickel or other metal plate submerged into the same solution. Metal ions would transfer from the plate through the solution into the membrane, producing high fidelity metal replica of the grating on the membrane. In one variation, an adhesive may be deposited on the deformable substrate, and then cured without touching the rigid, curved substrate. Edges of the deformable substrate may be attached to the rigid substrate to ensure uniform deformation of the deformable substrate. The assembly may be performed in vacuum, and then taken out to atmospheric pressure conditions to ensure that no air is trapped between the deformable and rigid substrates. Alternatively, a rigid surface with complementary curvature to the rigid substrate may be used to ensure uniform adhesion of the deformable substrate to the rigid substrate. Liquid may be applied to the surface of the deformable substrate to uniformly distribute pressure across its surface during the curing or hardening of the adhesive, or the film may be pressed into the surface using a deformable object or surface. After the attachment is complete, the grooves may be coated with reflective or dielectric layers to improve diffraction efficiency.

Apical constriction drives tissue-scale hydrodynamic flow to mediate cell elongation

PubMed Central

He, Bing; Doubrovinski, Konstantin; Polyakov, Oleg; Wieschaus, Eric

2014-01-01

Epithelial folding mediated by apical constriction converts flat epithelial sheets into multilayered, complex tissue structures and is employed throughout the development in most animals1. Little is known, however, how forces produced near the apical surface of the tissue are transmitted within individual cells to generate the global changes in cell shape that characterize tissue deformation. Here we apply particle tracking velocimetry in gastrulating Drosophila embryos to measure the movement of cytoplasm and plasma membrane during ventral furrow (VF) formation2, 3. We find that cytoplasmic redistribution during the lengthening phase of VF formation can be precisely described by viscous flows that quantitatively match the predictions of hydrodynamics. Cell membranes move with the ambient cytoplasm, with little resistance to or driving force on the flow. Strikingly, apical constriction produces similar flow patterns in mutant embryos that fail to form cells prior to gastrulation (“acellular” embryos), such that the global redistribution of cytoplasm mirrors the summed redistribution occurring in individual cells of wild type embryos. Our results suggest that during the lengthening phase of VF formation, hydrodynamic behavior of the cytoplasm provides the predominant mechanism transmitting apically generated forces deep into the tissue and that cell individualization is dispensable. PMID:24590071

A Microfluidic System with Surface Patterning for Investigating Cavitation Bubble(s)-Cell Interaction and the Resultant Bioeffects at the Single-cell Level.

PubMed

Li, Fenfang; Yuan, Fang; Sankin, Georgy; Yang, Chen; Zhong, Pei

2017-01-10

In this manuscript, we first describe the fabrication protocol of a microfluidic chip, with gold dots and fibronectin-coated regions on the same glass substrate, that precisely controls the generation of tandem bubbles and individual cells patterned nearby with well-defined locations and shapes. We then demonstrate the generation of tandem bubbles by using two pulsed lasers illuminating a pair of gold dots with a few-microsecond time delay. We visualize the bubble-bubble interaction and jet formation by high-speed imaging and characterize the resultant flow field using particle image velocimetry (PIV). Finally, we present some applications of this technique for single cell analysis, including cell membrane poration with macromolecule uptake, localized membrane deformation determined by the displacements of attached integrin-binding beads, and intracellular calcium response from ratiometric imaging. Our results show that a fast and directional jetting flow is produced by the tandem bubble interaction, which can impose a highly localized shear stress on the surface of a cell grown in close proximity. Furthermore, different bioeffects can be induced by altering the strength of the jetting flow by adjusting the standoff distance from the cell to the tandem bubbles.

Distributed microscopic actuation analysis of deformable plate membrane mirrors

NASA Astrophysics Data System (ADS)

Lu, Yifan; Yue, Honghao; Deng, Zongquan; Tzou, Hornsen

2018-02-01

To further reduce the areal density of optical mirrors used in space telescopes and other space-borne optical structures, the concept of flexible membrane deformable mirror has been proposed. Because of their high flexibility, poor stiffness and low damping properties, environmental excitations such as orbital maneuver, path changing, and non-uniform heating may induce unexpected vibrations and thus reduce working performance. Therefore, active vibration control is essential for these membrane mirrors. In this paper, two different mirror models, i.e., the plate membrane model and pure membrane model, are studied respectively. In order to investigate the modal vibration characteristics of the mirror, a piezoelectric layer is fully laminated on its non-reflective side to serve as actuators. Dynamic equations of the mirror laminated with piezoelectric actuators are presented first. Then, the actuator induced modal control force is defined. When the actuator area shrinks to infinitesimal, the expressions of microscopic local modal control force and its two components are obtained to predict the spatial microscopic actuation behavior of the mirror. Different membrane pretension forces are also applied to reveal the tension effects on the actuation of the mirror. Analyses indicate that the spatial distribution of modal micro-control forces is exactly the same with the sensing signals distribution of the mirror, which provides crucial guidelines for optimal actuator placement of membrane deformable mirrors.

Coupled biopolymer networks

NASA Astrophysics Data System (ADS)

Schwarz, J. M.; Zhang, Tao

2015-03-01

The actin cytoskeleton provides the cell with structural integrity and allows it to change shape to crawl along a surface, for example. The actin cytoskeleton can be modeled as a semiflexible biopolymer network that modifies its morphology in response to both external and internal stimuli. Just inside the inner nuclear membrane of a cell exists a network of filamentous lamin that presumably protects the heart of the cell nucleus--the DNA. Lamins are intermediate filaments that can also be modeled as semiflexible biopolymers. It turns out that the actin cytoskeletal biopolymer network and the lamin biopolymer network are coupled via a sequence of proteins that bridge the outer and inner nuclear membranes. We, therefore, probe the consequences of such a coupling via numerical simulations to understand the resulting deformations in the lamin network in response to perturbations in the cytoskeletal network. Such study could have implications for mechanical mechanisms of the regulation of transcription, since DNA--yet another semiflexible polymer--contains lamin-binding domains, and, thus, widen the field of epigenetics.

Compressive deformation of a single microcapsule

NASA Astrophysics Data System (ADS)

Liu, K. K.; Williams, D. R.; Briscoe, B. J.

1996-12-01

This paper reports an experimental and theoretical study of the compressive behavior of single microcapsules; that is, liquid-filled cellular entities (approximately 65 μm in diameter) with a thin polymeric membrane wall. An experimental technique which allows the simultaneous measurement of both the compressive displacement and the reaction forces of individual microcapsules deformed between two parallel plates up to a dimensionless approach [(compressive displacement)/(initial particle diameter)] of 60% is described. The corresponding major geometric parameters of the deformed microcapsule, such as central lateral extension as well as the failure phenomena, are reported and recorded through a microscopic visualization system. The elastic modulus, the bursting strength of the membrane, and the pressure difference across the membrane are computed by using a theoretical analysis which is also presented in this paper. This theoretical model, which was developed by Feng and Yang [

Deformations of a pre-stretched and lubricated finite elastic membrane driven by non-uniform external forcing

NASA Astrophysics Data System (ADS)

Boyko, Evgeniy; Gat, Amir; Bercovici, Moran

2017-11-01

We study viscous-elastic dynamics of a fluid confined between a rigid plate and a finite pre-stretched circular elastic membrane, pinned at its boundaries. The membrane is subjected to forces acting either directly on the membrane or through a pressure distribution in the fluid. Under the assumptions of strong pre-stretching and small deformations of the elastic sheet, and by applying the lubrication approximation for the flow, we derive the Green's function for the resulting linearized 4th order diffusion equation governing the deformation field in cylindrical coordinates. In addition, defining an asymptotic expansion with the ratio of the induced to prescribed tension serving as the small parameter, we reduce the coupled Reynolds and non-linear von-Karman equations to a set of three one-way coupled linear equations. The solutions to these equations provide insight onto the effects of induced tension, and enable simplified prediction of the correction for the deformation field. Funded by the European Research Council (ERC) under the European Union'sHorizon 2020 Research and Innovation Programme, Grant Agreement No. 678734 (MetamorphChip). E.B. is supported by the Adams Fellowship Program.

Review of infrared scene projector technology-1993

NASA Astrophysics Data System (ADS)

Driggers, Ronald G.; Barnard, Kenneth J.; Burroughs, E. E.; Deep, Raymond G.; Williams, Owen M.

1994-07-01

The importance of testing IR imagers and missile seekers with realistic IR scenes warrants a review of the current technologies used in dynamic infrared scene projection. These technologies include resistive arrays, deformable mirror arrays, mirror membrane devices, liquid crystal light valves, laser writers, laser diode arrays, and CRTs. Other methods include frustrated total internal reflection, thermoelectric devices, galvanic cells, Bly cells, and vanadium dioxide. A description of each technology is presented along with a discussion of their relative benefits and disadvantages. The current state of each methodology is also summarized. Finally, the methods are compared and contrasted in terms of their performance parameters.

On the Mechanism of Human Red Blood Cell Longevity: Roles of Calcium, the Sodium Pump, PIEZO1, and Gardos Channels.

PubMed

Lew, Virgilio L; Tiffert, Teresa

2017-01-01

In a healthy adult, the transport of O 2 and CO 2 between lungs and tissues is performed by about 2 · 10 13 red blood cells, of which around 1.7 · 10 11 are renewed every day, a turnover resulting from an average circulatory lifespan of about 120 days. Cellular lifespan is the result of an evolutionary balance between the energy costs of maintaining cells in a fit functional state versus cell renewal. In this Review we examine how the set of passive and active membrane transporters of the mature red blood cells interact to maximize their circulatory longevity thus minimizing costs on expensive cell turnover. Red blood cell deformability is critical for optimal rheology and gas exchange functionality during capillary flow, best fulfilled when the volume of each human red blood cell is kept at a fraction of about 0.55-0.60 of the maximal spherical volume allowed by its membrane area, the optimal-volume-ratio range. The extent to which red blood cell volumes can be preserved within or near these narrow optimal-volume-ratio margins determines the potential for circulatory longevity. We show that the low cation permeability of red blood cells allows volume stability to be achieved with extraordinary cost-efficiency, favouring cell longevity over cell turnover. We suggest a mechanism by which the interplay of a declining sodium pump and two passive membrane transporters, the mechanosensitive PIEZO1 channel, a candidate mediator of P sickle in sickle cells, and the Ca 2+ -sensitive, K + -selective Gardos channel, can implement red blood cell volume stability around the optimal-volume-ratio range, as required for extended circulatory longevity.

On the Mechanism of Human Red Blood Cell Longevity: Roles of Calcium, the Sodium Pump, PIEZO1, and Gardos Channels

PubMed Central

Lew, Virgilio L.; Tiffert, Teresa

2017-01-01

In a healthy adult, the transport of O2 and CO2 between lungs and tissues is performed by about 2 · 1013 red blood cells, of which around 1.7 · 1011 are renewed every day, a turnover resulting from an average circulatory lifespan of about 120 days. Cellular lifespan is the result of an evolutionary balance between the energy costs of maintaining cells in a fit functional state versus cell renewal. In this Review we examine how the set of passive and active membrane transporters of the mature red blood cells interact to maximize their circulatory longevity thus minimizing costs on expensive cell turnover. Red blood cell deformability is critical for optimal rheology and gas exchange functionality during capillary flow, best fulfilled when the volume of each human red blood cell is kept at a fraction of about 0.55–0.60 of the maximal spherical volume allowed by its membrane area, the optimal-volume-ratio range. The extent to which red blood cell volumes can be preserved within or near these narrow optimal-volume-ratio margins determines the potential for circulatory longevity. We show that the low cation permeability of red blood cells allows volume stability to be achieved with extraordinary cost-efficiency, favouring cell longevity over cell turnover. We suggest a mechanism by which the interplay of a declining sodium pump and two passive membrane transporters, the mechanosensitive PIEZO1 channel, a candidate mediator of Psickle in sickle cells, and the Ca2+-sensitive, K+-selective Gardos channel, can implement red blood cell volume stability around the optimal-volume-ratio range, as required for extended circulatory longevity. PMID:29311949

Continuum modeling of neuronal cell under blast loading

PubMed Central

Jérusalem, Antoine; Dao, Ming

2012-01-01

Traumatic brain injuries have recently been put under the spotlight as one of the most important causes of accidental brain dysfunctions. Significant experimental and modeling efforts are thus ongoing to study the associated biological, mechanical and physical mechanisms. In the field of cell mechanics, progresses are also being made at the experimental and modeling levels to better characterize many of the cell functions such as differentiation, growth, migration and death, among others. The work presented here aims at bridging both efforts by proposing a continuum model of neuronal cell submitted to blast loading. In this approach, cytoplasm, nucleus and membrane (plus cortex) are differentiated in a representative cell geometry, and different material constitutive models are adequately chosen for each one. The material parameters are calibrated against published experimental work of cell nanoindentation at multiple rates. The final cell model is ultimately subjected to blast loading within a complete fluid-structure interaction computational framework. The results are compared to the nanoindentation simulation and the specific effects of the blast wave on the pressure and shear levels at the interfaces are identified. As a conclusion, the presented model successfully captures some of the intrinsic intracellular phenomena occurring during its deformation under blast loading and potentially leading to cell damage. It suggests more particularly the localization of damage at the nucleus membrane similarly to what has already been observed at the overall cell membrane. This degree of damage is additionally predicted to be worsened by a longer blast positive phase duration. As a conclusion, the proposed model ultimately provides a new three dimensional computational tool to evaluate intracellular damage during blast loading. PMID:22562014

Nuclear pore assembly proceeds by an inside-out extrusion of the nuclear envelope

PubMed Central

Otsuka, Shotaro; Bui, Khanh Huy; Schorb, Martin; Hossain, M Julius; Politi, Antonio Z; Koch, Birgit; Eltsov, Mikhail; Beck, Martin; Ellenberg, Jan

2016-01-01

The nuclear pore complex (NPC) mediates nucleocytoplasmic transport through the nuclear envelope. How the NPC assembles into this double membrane boundary has remained enigmatic. Here, we captured temporally staged assembly intermediates by correlating live cell imaging with high-resolution electron tomography and super-resolution microscopy. Intermediates were dome-shaped evaginations of the inner nuclear membrane (INM), that grew in diameter and depth until they fused with the flat outer nuclear membrane. Live and super-resolved fluorescence microscopy revealed the molecular maturation of the intermediates, which initially contained the nuclear and cytoplasmic ring component Nup107, and only later the cytoplasmic filament component Nup358. EM particle averaging showed that the evagination base was surrounded by an 8-fold rotationally symmetric ring structure from the beginning and that a growing mushroom-shaped density was continuously associated with the deforming membrane. Quantitative structural analysis revealed that interphase NPC assembly proceeds by an asymmetric inside-out extrusion of the INM. DOI: http://dx.doi.org/10.7554/eLife.19071.001 PMID:27630123

Sensor for Injection Rate Measurements

PubMed Central

Marcic, Milan

2006-01-01

A vast majority of the medium and high speed Diesel engines are equipped with multi-hole injection nozzles nowadays. Inaccuracies in workmanship and changing hydraulic conditions in the nozzles result in differences in injection rates between individual injection nozzle holes. The new deformational measuring method described in the paper allows injection rate measurement in each injection nozzle hole. The differences in injection rates lead to uneven thermal loads of Diesel engine combustion chambers. All today known measuring method, such as Bosch and Zeuch give accurate results of the injection rate in diesel single-hole nozzles. With multihole nozzles they tell us nothing about possible differences in injection rates between individual holes of the nozzle. At deformational measuring method, the criterion of the injected fuel is expressed by the deformation of membrane occurring due to the collision of the pressure wave against the membrane. The pressure wave is generated by the injection of the fuel into the measuring space. For each hole of the nozzle the measuring device must have a measuring space of its own into which fuel is injected as well as its measuring membrane and its own fuel outlet. During measurements procedure the measuring space must be filled with fuel to maintain an overpressure of 5 kPa. Fuel escaping from the measuring device is conducted into the graduated cylinders for measuring the volumetric flow through each hole of the nozzle.The membrane deformation is assessed by strain gauges. They are glued to the membrane and forming the full Wheatstone's bridge. We devoted special attention to the membrane shape and temperature compensation of the strain gauges.

Spectrin tetramer-dimer equilibrium and the stability of erythrocyte membrane skeletons

NASA Astrophysics Data System (ADS)

Liu, Shih-Chun; Palek, Jiri

1980-06-01

The inner side of the red-cell membrane is laminated by a two-dimensional network of membrane proteins which include spectrin, actin and some other components1-4. After extraction of lipids and integral proteins from the membrane, this membrane skeleton can be visualized as a ball-shaped network consisting of twisted fibres1-4 and globular protrusions4; however, the assembly of the individual proteins in the membrane skeleton is not well understood. Spectrin can be eluted from the membrane in the form of dimers and tetramers5-8. Electron microscopic study with low-angle shadowing technique shows that spectrin dimers are two parallel strands of twisted fibres presumably representing bands 1 and 2 of spectrin9. Spectrin tetramers presumably formed by head-to-head associations of two dimers are twice as long9. In solution, the spectrin dimer-tetramer equilibrium depends on temperature and salt concentration7,8; however, it is not known whether the same equilibrium exists in the membrane and whether it affects the physical properties of the membrane, such as its structural stability and deformability. We now demonstrate that spectrin dimers and tetramers are in a reversible equilibrium in the membrane and that in physiological conditions this equilibrium favours spectrin tetramers. Furthermore, we show that transformation of spectrin tetramers to dimers, as induced by ghost incubation in hypotonic conditions, diminishes the structural stability of the Triton-insoluble membrane skeletons.

46 CFR 154.427 - Membrane tank system design.

Code of Federal Regulations, 2010 CFR

2010-10-01

... 46 Shipping 5 2010-10-01 2010-10-01 false Membrane tank system design. 154.427 Section 154.427... Membrane Tanks § 154.427 Membrane tank system design. A membrane tank system must be designed for: (a) Any static and dynamic loads with respect to plastic deformation and fatigue; (b) Combined strains from...

In vivo influence of extract from Aronia melanocarpa on the erythrocyte membranes in patients with hypercholesterolemia.

PubMed

Duchnowicz, Piotr; Nowicka, Agmieszka; Koter-Michalak, Maria; Broncel, Marlena

2012-09-01

Hypercholesterolemia increases cholesterol concentration in erythrocyte membranes, which results in decrease of membrane fluidity and decreases the deformability of red blood cells. The fruits of Arona melanocarpa contains many of polyphenols and other compounds that have beneficial health effects. The aim of the study was to estimate the influence of 2-month supplementation of extract from Aronia melanocarpa (100 mg Aronox, three times per day) on cholesterol concentration, lipid peroxidation, membrane fluidity, level of thiol groups and activity of ATPase in erythrocytes from patients with hypercholesterolemia. The study involved 25 patients with hypercholesterolemia without pharmacological treatment and 20 healthy individuals as a control group. Blood samples were collected before, and after 1 and 2 months of Aronia administration. The 2-month Aronia supplementation resulted in a decrease of cholesterol concentration (by 22%) and a decrease of lipid peroxidation (by 40%), and an increase of membrane fluidity. No statistically significant increase of the concentration of thiol groups and of ATPase activity were observed. Our study shows that supplementation of extract from Aronia melanocarpa has a beneficial effect on rheological properties of erythrocytes.

Probing eukaryotic cell mechanics via mesoscopic simulations

PubMed Central

Shang, Menglin; Lim, Chwee Teck

2017-01-01

Cell mechanics has proven to be important in many biological processes. Although there is a number of experimental techniques which allow us to study mechanical properties of cell, there is still a lack of understanding of the role each sub-cellular component plays during cell deformations. We present a new mesoscopic particle-based eukaryotic cell model which explicitly describes cell membrane, nucleus and cytoskeleton. We employ Dissipative Particle Dynamics (DPD) method that provides us with the unified framework for modeling of a cell and its interactions in the flow. Data from micropipette aspiration experiments were used to define model parameters. The model was validated using data from microfluidic experiments. The validated model was then applied to study the impact of the sub-cellular components on the cell viscoelastic response in micropipette aspiration and microfluidic experiments. PMID:28922399

Exploiting Conjugated Polyelectrolyte Photophysics toward Monitoring Real-Time Lipid Membrane-Surface Interaction Dynamics at the Single-Particle Level.

PubMed

Calver, Christina F; Liu, Hsiao-Wei; Cosa, Gonzalo

2015-11-03

Herein we report the real-time observation of the interaction dynamics between cationic liposomes flowing in solution and a surface-immobilized charged scaffolding formed by the deposition of conjugated polyanion poly[5-methoxy-2-(3-sulfopropoxy)-1,4-phenylenevinylene (MPS-PPV) onto 100-nm-diameter SiO2 nanoparticles (NPs). Contact of the freely floating liposomes with the polymer-coated surfaces led to the formation of supported lipid bilayers (SLBs). The interaction of the incoming liposomes with MPS-PPV adsorbed on individual SiO2 nanoparticles promoted the deaggregation of the polymer conformation and led to large emission intensity enhancements. Single-particle total internal reflection fluorescence microscopy studies exploited this phenomenon as a way to monitor the deformation dynamics of liposomes on surface-immobilized NPs. The MPS-PPV emission enhancement (up to 25-fold) reflected on the extent of membrane contact with the surface of the NP and was correlated with the size of the incoming liposome. The time required for the MPS-PPV emission to reach a maximum (ranging from 400 to 1000 ms) revealed the dynamics of membrane deformation and was also correlated with the liposome size. Cryo-TEM experiments complemented these results by yielding a structural view of the process. Immediately following the mixing of liposomes and NPs the majority of NPs had one or more adsorbed liposomes, yet the presence of a fully formed SLB was rare. Prolonged incubation of liposomes and NPs showed completely formed SLBs on all of the NPs, confirming that the liposomes eventually ruptured to form SLBs. We foresee that the single-particle studies we report herein may be readily extended to study membrane dynamics of other lipids including cellular membranes in live cell studies and to monitor the formation of polymer-cushioned SLBs.

Nitric oxide in red blood cell adaptation to hypoxia.

PubMed

Zhao, Yajin; Wang, Xiang; Noviana, Milody; Hou, Man

2018-06-01

Nitric oxide (NO) appears to be involved in virtually every aspect of cardiovascular biology. Most attention has been focused on the role of endothelial-derived NO in basal blood flow regulation by relaxing vascular smooth muscle; however, it is now known that NO derived from red blood cells (RBCs) plays a fundamental role in vascular homeostasis by enhancing oxygen (O2) release at the cellular and physiological level. Hypoxia is an often seen problem in diverse conditions; systemic adaptations to hypoxia permit people to adjust to the hypoxic environment at high altitudes and to disease processes. In addition to the cardiopulmonary and hematologic adaptations that support systemic O2 delivery in hypoxia, RBCs assist through newly described NO-based mechanisms, in line with their vital role in O2 transport and delivery. Furthermore, to increase the local blood flow in proportion to metabolic demand, NO regulates membrane mechanical properties thereby modulating RBC deformability and O2 carrying-releasing function. In this review article, we focus on the effect of NO bioactivity on RBC-based mechanisms that regulate blood flow and RBC deformability. RBC adaptations to hypoxia are summarized, with particular attention to NO-dependent S-nitrosylation of membrane proteins and hemoglobin (S-nitrosohemoglobin). The NO/S-nitrosylation/RBC vasoregulatory cascade contributes fundamentally to the molecular understanding of the role of NO in human adaptation to hypoxia and may inform novel therapeutic strategies.

Molecular structure and differential function of choline kinases CHKα and CHKβ in musculoskeletal system and cancer.

PubMed

Chen, Xi; Qiu, Heng; Wang, Chao; Yuan, Yu; Tickner, Jennifer; Xu, Jiake; Zou, Jun

2017-02-01

Choline, a hydrophilic cation, has versatile physiological roles throughout the body, including cholinergic neurotransmission, memory consolidation and membrane biosynthesis and metabolism. Choline kinases possess enzyme activity that catalyses the conversion of choline to phosphocholine, which is further converted to cytidine diphosphate-coline (CDP-choline) in the biosynthesis of phosphatidylcholine (PC). PC is a major constituent of the phospholipid bilayer which constitutes the eukaryotic cell membrane, and regulates cell signal transduction. Choline Kinase consists of three isoforms, CHKα1, CHKα2 and CHKβ, encoded by two separate genes (CHKA(Human)/Chka(Mouse) and CHKB(Human)/Chkb(Mouse)). Both isoforms have similar structures and enzyme activity, but display some distinct molecular structural domains and differential tissue expression patterns. Whilst Choline Kinase was discovered in early 1950, its pivotal role in the development of muscular dystrophy, bone deformities, and cancer has only recently been identified. CHKα has been proposed as a cancer biomarker and its inhibition as an anti-cancer therapy. In contrast, restoration of CHKβ deficiency through CDP-choline supplements like citicoline may be beneficial for the treatment of muscular dystrophy, bone metabolic diseases, and cognitive conditions. The molecular structure and expression pattern of Choline Kinase, the differential roles of Choline Kinase isoforms and their potential as novel therapeutic targets for muscular dystrophy, bone deformities, cognitive conditions and cancer are discussed. Copyright © 2016. Published by Elsevier Ltd.

Microfluidic Investigation Reveals Distinct Roles for Actin Cytoskeleton and Myosin II Activity in Capillary Leukocyte Trafficking

PubMed Central

Gabriele, Sylvain; Benoliel, Anne-Marie; Bongrand, Pierre; Théodoly, Olivier

2009-01-01

Circulating leukocyte sequestration in pulmonary capillaries is arguably the initiating event of lung injury in acute respiratory distress syndrome. We present a microfluidic investigation of the roles of actin organization and myosin II activity during the different stages of leukocyte trafficking through narrow capillaries (entry, transit and shape relaxation) using specific drugs (latrunculin A, jasplakinolide, and blebbistatin). The deformation rate during entry reveals that cell stiffness depends strongly on F-actin organization and hardly on myosin II activity, supporting a microfilament role in leukocyte sequestration. In the transit stage, cell friction is influenced by stiffness, demonstrating that the actin network is not completely broken after a forced entry into a capillary. Conversely, membrane unfolding was independent of leukocyte stiffness. The surface area of sequestered leukocytes increased by up to 160% in the absence of myosin II activity, showing the major role of molecular motors in microvilli wrinkling and zipping. Finally, cell shape relaxation was largely independent of both actin organization and myosin II activity, whereas a deformed state was required for normal trafficking through capillary segments. PMID:19450501

Adaptive metal mirror for high-power CO2 lasers

NASA Astrophysics Data System (ADS)

Jarosch, Uwe-Klaus

1996-08-01

Spherical mirrors with a variable radius of curvature are used inside laser resonators as well as in the beam path between the laser and the workpiece. Commercially-available systems use piezoelectric actuators, or the pressure of the coolant, to deform the mirror surface. In both cases, the actuator and the cooling system influence each other. This interaction is avoided through the integration of the cooling system with the flexible mirror membrane. A multi- channel design leads to an optimized cooling effect, which is necessary for high power applications. The contour of the variable metal mirror depends on the mounting between the membrane and the mirror body and on the distribution of forces. Four cases of deformation can be distinguished for a circular elastic membrane. The realization of an adaptive metal mirror requires a technical compromise to be made. A mechanical construction is presented which combines an elastic hinge with the inlet and outlet of the coolant. For the deformation of the mirror membranes two actuators with different character of deformation are used. The superposition of the two deformations results in smaller deviations from the spherical surface shape than can be achieved using a single actuator. DC proportional magnets have been introduced as cheap and rigid actuators. The use of this adaptive mirror, either in a low pressure atmosphere of a gas laser resonator, or in an extra-cavity beam path is made possible through the use of a ventilation system.

A biochemo-mechano coupled, computational model combining membrane transport and pericellular proteolysis in tissue mechanics

PubMed Central

Vuong, A.-T.; Rauch, A. D.

2017-01-01

We present a computational model for the interaction of surface- and volume-bound scalar transport and reaction processes with a deformable porous medium. The application in mind is pericellular proteolysis, i.e. the dissolution of the solid phase of the extracellular matrix (ECM) as a response to the activation of certain chemical species at the cell membrane and in the vicinity of the cell. A poroelastic medium model represents the extra cellular scaffold and the interstitial fluid flow, while a surface-bound transport model accounts for the diffusion and reaction of membrane-bound chemical species. By further modelling the volume-bound transport, we consider the advection, diffusion and reaction of sequestered chemical species within the extracellular scaffold. The chemo-mechanical coupling is established by introducing a continuum formulation for the interplay of reaction rates and the mechanical state of the ECM. It is based on known experimental insights and theoretical work on the thermodynamics of porous media and degradation kinetics of collagen fibres on the one hand and a damage-like effect of the fibre dissolution on the mechanical integrity of the ECM on the other hand. The resulting system of partial differential equations is solved via the finite-element method. To the best of our knowledge, it is the first computational model including contemporaneously the coupling between (i) advection–diffusion–reaction processes, (ii) interstitial flow and deformation of a porous medium, and (iii) the chemo-mechanical interaction impelled by the dissolution of the ECM. Our numerical examples show good agreement with experimental data. Furthermore, we outline the capability of the methodology to extend existing numerical approaches towards a more comprehensive model for cellular biochemo-mechanics. PMID:28413347

Rheological behavior of rat mesangial cells during swelling in vitro.

PubMed

Craelius, W; Huang, C J; Guber, H; Palant, C E

1997-01-01

The response of cells to mechanical forces depends on the rheological properties of their membranes and cytoplasm. To characterize those properties, mechanical and electrical responses to swelling were measured in rat mesangial cells (MC) using electrophysiologic and video microscopic techniques. Ion transport rates during hyposmotic exposures were measured with whole-cell recording electrodes. Results showed that cell swelling varied nonlinearly with positive internal pressure, consistent with a viscoelastic cytoplasm. The extrapolated area expansivity modulus for small deformations was estimated to be 450 dyne/cm. Cell swelling, caused either by positive pipet pressure or hyposmotic exposure (40-60 mOsm Kg-1), rapidly induced an outwardly rectifying membrane conductance with an outward magnitude 4-5 times the baseline conductance of 0.9 +/- 0.5 nS (p < .01). Swelling-induced (SI) current was weakly selective for K+ over Na+, partially reversed upon return to isotonicity, and was antagonized by 0.5 mM GdCl3 (p < 0.02; n = 6). Isolated cells treated with GdCl3 rapidly lysed after hypotonic exposure, in contrast to untreated cells that exhibited regulatory volume decrease (RVD). Our results indicate that volume regulation by MC depends upon a large swelling-induced K+ efflux, and suggest that swelling in MC is a viscoelastic process, with a viscosity dependent on the degree of swelling.

Micrometer-Scale Membrane Transition of Supported Lipid Bilayer Membrane Reconstituted with Cytosol of Dictyostelium discoideum.

PubMed

Takahashi, Kei; Toyota, Taro

2017-03-07

The transformation of the supported lipid bilayer (SLB) membrane by extracted cytosol from living resources, has recently drawn much attention. It enables us to address the question of whether the purified phospholipid SLB membrane, including lipids related to amoeba locomotion, which was discussed in many previous studies, exhibits membrane deformation in the presence of cytosol extracted from amoeba; Methods: In this report, a method for reconstituting a supported lipid bilayer (SLB) membrane, composed of purified phospholipids and cytosol extracted from Dictyostelium discoideum , is described. This technique is a new reconstitution method combining the artificial constitution of membranes with the reconstitution using animate cytosol (without precise purification at a molecular level), contributing to membrane deformation analysis; Results: The morphology transition of a SLB membrane composed of phosphatidylcholines, after the addition of cytosolic extract, was traced using a confocal laser scanning fluorescence microscope. As a result, pore formation in the SLB membrane was observed and phosphatidylinositides incorporated into the SLB membrane tended to suppress pore formation and expansion; Conclusions: The current findings imply that phosphatidylinositides have the potential to control cytoplasm activity and bind to a phosphoinositide-containing SLB membrane.

Free energy analysis of cell spreading.

PubMed

McEvoy, Eóin; Deshpande, Vikram S; McGarry, Patrick

2017-10-01

In this study we present a steady-state adaptation of the thermodynamically motivated stress fiber (SF) model of Vigliotti et al. (2015). We implement this steady-state formulation in a non-local finite element setting where we also consider global conservation of the total number of cytoskeletal proteins within the cell, global conservation of the number of binding integrins on the cell membrane, and adhesion limiting ligand density on the substrate surface. We present a number of simulations of cell spreading in which we consider a limited subset of the possible deformed spread-states assumed by the cell in order to examine the hypothesis that free energy minimization drives the process of cell spreading. Simulations suggest that cell spreading can be viewed as a competition between (i) decreasing cytoskeletal free energy due to strain induced assembly of cytoskeletal proteins into contractile SFs, and (ii) increasing elastic free energy due to stretching of the mechanically passive components of the cell. The computed minimum free energy spread area is shown to be lower for a cell on a compliant substrate than on a rigid substrate. Furthermore, a low substrate ligand density is found to limit cell spreading. The predicted dependence of cell spread area on substrate stiffness and ligand density is in agreement with the experiments of Engler et al. (2003). We also simulate the experiments of Théry et al. (2006), whereby initially circular cells deform and adhere to "V-shaped" and "Y-shaped" ligand patches. Analysis of a number of different spread states reveals that deformed configurations with the lowest free energy exhibit a SF distribution that corresponds to experimental observations, i.e. a high concentration of highly aligned SFs occurs along free edges, with lower SF concentrations in the interior of the cell. In summary, the results of this study suggest that cell spreading is driven by free energy minimization based on a competition between decreasing cytoskeletal free energy and increasing passive elastic free energy. Copyright © 2017 Elsevier Ltd. All rights reserved.

Large aperture deformable mirror with a transferred single-crystal silicon membrane actuated using large-stroke PZT Unimorph Actuators

NASA Technical Reports Server (NTRS)

Hishinumat, Yoshikazu; Yang, Eui - Hyeok (EH)

2005-01-01

We have demonstrated a large aperture (50 mm x 50 mm) continuous membrane deformable mirror (DM) with a large-stroke piezoelectric unimorph actuator array. The DM consists of a continuous, large aperture, silicon membrane 'transferred' in its entirety onto a 20 x 20 piezoelectric unimorph actuator array. A PZT unimorph actuator, 2.5 mm in diameter with optimized PZT/Si thickness and design showed a deflection of 5.7 [m at 20V. An assembled DM showed an operating frequency bandwidth of 30 kHz and influence function of approximately 30%.

Comparison of cell behavior on pva/pva-gelatin electrospun nanofibers with random and aligned configuration

NASA Astrophysics Data System (ADS)

Huang, Chen-Yu; Hu, Keng-Hsiang; Wei, Zung-Hang

2016-12-01

Electrospinning technique is able to create nanofibers with specific orientation. Poly(vinyl alcohol) (PVA) have good mechanical stability but poor cell adhesion property due to the low affinity of protein. In this paper, extracellular matrix, gelatin is incorporated into PVA solution to form electrospun PVA-gelatin nanofibers membrane. Both randomly oriented and aligned nanofibers are used to investigate the topography-induced behavior of fibroblasts. Surface morphology of the fibers is studied by optical microscopy and scanning electron microscopy (SEM) coupled with image analysis. Functional group composition in PVA or PVA-gelatin is investigated by Fourier Transform Infrared (FTIR). The morphological changes, surface coverage, viability and proliferation of fibroblasts influenced by PVA and PVA-gelatin nanofibers with randomly orientated or aligned configuration are systematically compared. Fibroblasts growing on PVA-gelatin fibers show significantly larger projected areas as compared with those cultivated on PVA fibers which p-value is smaller than 0.005. Cells on PVA-gelatin aligned fibers stretch out extensively and their intracellular stress fiber pull nucleus to deform. Results suggest that instead of the anisotropic topology within the scaffold trigger the preferential orientation of cells, the adhesion of cell membrane to gelatin have substantial influence on cellular behavior.

Spore formation in Myxococcus xanthus is tied to cytoskeleton functions and polysaccharide spore coat deposition

PubMed Central

Müller, Frank D.; Schink, Christian W.; Hoiczyk, Egbert; Cserti, Emöke; Higgs, Penelope I.

2011-01-01

Summary Myxococcus xanthus is a Gram-negative bacterium that differentiates into environmentally resistant spores. Spore differentiation involves septation-independent remodelling of the rod-shaped vegetative cell into a spherical spore and deposition of a thick and compact spore coat outside of the outer membrane. Our analyses suggest that spore coat polysaccharides are exported to the cell surface by the Exo outer membrane polysaccharide export/polysaccharide co-polymerase 2a (OPX/PCP-2a) machinery. Conversion of the capsule-like polysaccharide layer into a compact spore coat layer requires the Nfs proteins which likely form a complex in the cell envelope. Mutants in either nfs, exo, or two other genetic loci encoding homologs of polysaccharide synthesis enzymes, fail to complete morphogenesis from rods to spherical spores and instead produce a transient state of deformed cell morphology before reversion into typical rods. We additionally provide evidence that the cell cytoskeletal protein, MreB, plays an important role in rod to spore morphogenesis and for spore outgrowth. These studies provide evidence that this novel gram-negative differentiation process is tied to cytoskeleton functions and polysaccharide spore coat deposition. PMID:22188356

Capillary flow enhancement in rectangular polymer microchannels with a deformable wall.

PubMed

Anoop, R; Sen, A K

2015-07-01

We report the capillary flow enhancement in rectangular polymer microchannels, when one of the channel walls is a deformable polymer membrane. We provide detailed insight into the physics of elastocapillary interaction between the capillary flow and elastic membrane, which leads to significant improvements in capillary flow performance. As liquid flows by capillary action in such channels, the deformable wall deflects inwards due to the Young-Laplace pressure drop across the liquid meniscus. This, in turn, decreases the radius of curvature of the meniscus and increases the driving capillary pressure. A theoretical model is proposed to predict the resultant increase in filling speed and rise height, respectively, in deformable horizontal and vertical microchannels having large aspect ratios. A non-dimensional parameter J, which represents the ratio of the capillary force to the mechanical restoring force, is identified to quantify the elastocapillary effects in terms of the improvement in filling speed (for J>0.238) and the condition for channel collapse (J>1). The theoretical predictions show good agreement with experimental data obtained using deformable rectangular poly(dimethylsiloxane) microchannels. Both model predictions and experimental data show that over 15% improvement in the Washburn coefficient in horizontal channels, and over 30% improvement in capillary rise height in vertical channels, are possible prior to channel collapse. The proposed technique of using deformable membranes as channel walls is a viable method for capillary flow enhancement in microfluidic devices.

Hereditary spherocytosis and elliptocytosis associated with prosthetic heart valve replacement: rheological study of erythrocyte modifications.

PubMed

Caprari, Patrizia; Tarzia, Anna; Mojoli, Giorgio; Cianciulli, Paolo; Mannella, Emilio; Martorana, Maria Cristina

2009-04-01

The implantation of a prosthetic heart valve (HVP) in patients with hereditary spherocytosis (HS) and hereditary elliptocytosis (HE) is rare, and the changes in the structure and deformability of erythrocytes that follow implantation in these patients have been poorly described. In the present study, the erythrocytes in HS and HE patients with mechanical HVP were compared to the erythrocytes in patients with only congenital membrane defects, in terms of biochemical modifications and rheological behaviour. Integral and cytoskeletal erythrocyte membrane proteins were studied, and blood viscosity (shear rate/shear stress ratio), aggregation ratio [eta(1 s(-1))/eta(200 s(-1))], and red cell visco-elasticity were determined. Valve replacement with a mechanical prosthesis worsened anaemia and resulted in a change in haemolysis, from sub-clinical to evident. The rheological investigation of erythrocytes from HS patients confirmed the characteristic increased viscosity and aggregation ratio and the decreased deformability. The rheological behaviour of erythrocytes from patients with HVP showed a decrease in viscosity and an increase in elastic modulus. In these patients, the prosthesis seems to have induced traumatic damage to the erythrocyte membrane, leading to fragmentation and lysis, which in turn modified rheological parameters. The biochemical and rheological investigation allowed us to understand the clinical and haematological pictures of the patients and to describe the role played by different factors in haemolytic anaemia.

Haemato-protective influence of dietary fenugreek (Trigonella foenum-graecum L.) seeds is potentiated by onion (Allium cepa L.) in streptozotocin-induced diabetic rats.

PubMed

Pradeep, Seetur R; Srinivasan, Krishnapura

2018-02-01

We have recently reported the beneficial modulation of metabolic abnormalities and oxidative stress in diabetic rats by dietary fenugreek seeds and onion. This investigation evaluated the protective influence of dietary fenugreek seeds (100 g kg -1 ) and onion (30 g kg -1 ) on erythrocytes of streptozotocin-induced diabetic rats, through modulation of reduced haematological indices and antisickling potency. This study also evaluated the altered erythrocyte membrane lipid profile and beneficial countering of increased lipid peroxidation, osmotic fragility, along with reduced membrane fluidity and deformability, nitric oxide production and echinocyte formation. Dietary fenugreek seeds and onion appeared to counter the deformity and fragility of erythrocytes partially in diabetic rats by their antioxidant potential and hypocholesterolemic property. The antisickling potency of these spices was accomplished by a substantial decrease in echinocyte population and AGEs in diabetic rats. Further insight into the factors that might have reduced the fluidity of erythrocytes in diabetic rats revealed changes in the cholesterol: phospholipid ratio, fatty acid profile, and activities of membrane-bound enzymes. Dietary fenugreek seeds and onion offered a beneficial protective effect to the red blood cells, the effect being higher with fenugreek + onion. This is the first report on the hemato-protective influence of a nutraceutical food component in diabetic situation. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

[In vitro anti-Trichomonas vaginalis effects of a mixture of dihydroartemisinin and metronidazole].

PubMed

Tang, Zi-Hao; Liu, Ke-Yue; Mei, Jun; Gao, Xing-Zheng

2010-12-30

To observe the effect of a mixture of dihydroartemisinin and metronidazole on ultrastructure of Trichomonas vaginalis trophozoites in vitro for exploring trichomonacidal mechanism of the drug mixture. The trophozoites were cultivated with liver extract solution medium that contained 2.5 x 10(6) parasites/ml. There were dihydroartemisinin 0.5 mg/ml and metronidazole 0.002 mg/ml in the experimental tubes of the drug mixture group. Groups of control (without drug), dihydroartemisinin (1 mg/ml) and metronidazole (5 mg/ml) were established and performed in the same experimental conditions. The parasites were observed by scanning and transmission electron microscopes after having treated with the drugs at 37 degrees for 3.5-5 h. Under scanning electron microscope, the cell membrane of T. vaginalis treated only with dihydroartemisinin for 35 h was damaged, part of pellicle peeled off. Although the surface of the trophozoites treated only with metronidazole for 5 h showed many small bubbles and hollows, the cell membrane looked integral. However, surface of the parasite exposed to the drug mixture for 3.5-4.2 h showed deep folds and cracks, the cell membrane was damaged and even peeled off. When the cell ruptured, the nucleus, axostyle, pelta and hydrogenosomes were exposed, and the cytoplasm spilled out. Transmission electron microscopy showed that the membrane system of the trophozoites treated only with dihydroartemisinin for 3.5 h was damaged considerably. The cytoplasm of damaged parasite spilled out. The cytoplasm of the parasite treated only with metronidazole for 3.5-5 h was damaged seriously. Vacuoles and crevices were visible in the cytoplasm. The cell membrane and the content of the parasites treated with the drug mixture for 3.5-4.5 h were damaged seriously. There were some vacuoles and crevices, dilated endoplasmic reticulum, injured and deformed hydrogenosomes in the cytoplasm. The cell organelles mostly disappeared. Crevices also existed in the nucleus. The nuclear membrane fractured and even disappeared. The acting targets of dihydroartemisinin and metronidazole to T. vaginalis trophozoite were different, and a combination of the two drugs shows stronger effect in killing the parasites.

Thermal and active fluctuations of a compressible bilayer vesicle

NASA Astrophysics Data System (ADS)

Sachin Krishnan, T. V.; Yasuda, Kento; Okamoto, Ryuichi; Komura, Shigeyuki

2018-05-01

We discuss thermal and active fluctuations of a compressible bilayer vesicle by using the results of hydrodynamic theory for vesicles. Coupled Langevin equations for the membrane deformation and the density fields are employed to calculate the power spectral density matrix of membrane fluctuations. Thermal contribution is obtained by means of the fluctuation dissipation theorem, whereas active contribution is calculated from exponentially decaying time correlation functions of active random forces. We obtain the total power spectral density as a sum of thermal and active contributions. An apparent response function is further calculated in order to compare with the recent microrheology experiment on red blood cells. An enhanced response is predicted in the low-frequency regime for non-thermal active fluctuations.

Mechanical properties of stored red blood cells using optical tweezers

NASA Astrophysics Data System (ADS)

Fontes, Adriana; Alexandre de Thomaz, Andre; de Ysasa Pozzo, Liliana; de Lourdes Barjas-Castro, Maria; Brandao, Marcelo M.; Saad, Sara T. O.; Barbosa, Luiz Carlos; Cesar, Carlos Lenz

2005-08-01

We have developed a method for measuring the red blood cell (RBC) membrane overall elasticity μ by measuring the deformation of the cells when dragged at a constant velocity through a plasma fluid by an optical tweezers. The deformability of erythrocytes is a critical determinant of blood flow in the microcirculation. We tested our method and hydrodynamic models, which included the presence of two walls, by measuring the RBC deformation as a function of drag velocity and of the distance to the walls. The capability and sensitivity of this method can be evaluated by its application to a variety of studies, such as, the measurement of RBC elasticity of sickle cell anemia patients comparing homozygous (HbSS), including patients taking hydroxyrea (HU) and heterozygous (HbAS) with normal donors and the RBC elasticity measurement of gamma irradiated stored blood for transfusion to immunosupressed patients as a function of time and dose. These studies show that the technique has the sensitivity to discriminate heterozygous and homozygous sickle cell anemia patients from normal donors and even follow the course of HU treatment of Homozygous patients. The gamma irradiation studies show that there is no significant change in RBC elasticity over time for up to 14 days of storage, regardless of whether the unit was irradiated or not, but there was a huge change in the measured elasticity for the RBC units stored for more than 21 days after irradiation. These finds are important for the assessment of stored irradiated RBC viability for transfusion purposes because the present protocol consider 28 storage days after irradiation as the limit for the RBC usage.

Inhibition of free radical scavenging enzymes affects mitochondrial membrane permeability transition during growth and aging of yeast cells.

PubMed

Deryabina, Yulia; Isakova, Elena; Sekova, Varvara; Antipov, Alexey; Saris, Nils-Erik L

2014-12-01

In this study, we investigated the change in the antioxidant enzymes activity, cell respiration, reactive oxygen species (ROS), and impairment of membrane mitochondria permeability in the Endomyces magnusii yeasts during culture growth and aging. We showed that the transition into stationary phase is the key tool to understanding interaction of these processes. This growth stage is distinguished by two-fold increase in ROS production and respiration rate as compared to those in the logarithmic phase. It results in induction of alternative oxidase (AO) in the stationary phase, decline of the main antioxidant enzymes activities, ROS-production, and mitochondria membrane permeability. Significant increase in the share of mitochondrial isoform of superoxide dismutase (SOD2) occurred in the stationary phase from 51.8% (24 h of cultivation) to 68.6% (48 h of cultivation). Upon blocking the essential ROS-scavenging enzymes, SODs and catalases (CATs) some heterogeneity of cell population was observed: 80-90% of cells displayed evident signs of early apoptosis (such as disorientation of mitochondria cristae, mitochondrial fragmentation and deformation of nuclear chromatine). However, 10-20% of the population were definitely healthy. It allowed to draw the conclusion that a complete system of cell antioxidant protection underlies normal mitochondria functioning while the E. magnusii yeasts grow and age. Moreover, this system provides unimpaired cell physiology under oxidative stress during culture aging in the stationary phase. Failures in mitochondria functions due to inhibition of ROS-scavenging enzymes of CATs and SODs could lead to damage of the cells and some signs of early apoptosis.

Coarse-grained Brownian ratchet model of membrane protrusion on cellular scale.

PubMed

Inoue, Yasuhiro; Adachi, Taiji

2011-07-01

Membrane protrusion is a mechanochemical process of active membrane deformation driven by actin polymerization. Previously, Brownian ratchet (BR) was modeled on the basis of the underlying molecular mechanism. However, because the BR requires a priori load that cannot be determined without information of the cell shape, it cannot be effective in studies in which resultant shapes are to be solved. Other cellular-scale models describing the protrusion have also been suggested for modeling a whole cell; however, these models were not developed on the basis of coarse-grained physics representing the underlying molecular mechanism. Therefore, to express the membrane protrusion on the cellular scale, we propose a novel mathematical model, the coarse-grained BR (CBR), which is derived on the basis of nonequilibrium thermodynamics theory. The CBR can reproduce the BR within the limit of the quasistatic process of membrane protrusion and can estimate the protrusion velocity consistently with an effective elastic constant that represents the state of the energy of the membrane. Finally, to demonstrate the applicability of the CBR, we attempt to perform a cellular-scale simulation of migrating keratocyte in which the proposed CBR is used for the membrane protrusion model on the cellular scale. The results show that the experimentally observed shapes of the leading edge are well reproduced by the simulation. In addition, The trend of dependences of the protrusion velocity on the curvature of the leading edge, the temperature, and the substrate stiffness also agreed with the other experimental results. Thus, the CBR can be considered an appropriate cellular-scale model to express the membrane protrusion on the basis of its underlying molecular mechanism.

A Microfluidic System with Surface Patterning for Investigating Cavitation Bubble(s)-Cell Interaction and the Resultant Bioeffects at the Single-Cell Level

PubMed Central

Li, Fenfang; Yuan, Fang; Sankin, Georgy; Yang, Chen; Zhong, Pei

2017-01-01

In this manuscript, we first describe the fabrication protocol of a microfluidic chip, with gold dots and fibronectin-coated regions on the same glass substrate that precisely controls the generation of tandem bubbles and individual cells patterned nearby with well-defined locations and shapes. We then demonstrate the generation of tandem bubbles by using two pulsed lasers illuminating a pair of gold dots with a few-microsecond time delay. We visualize the bubble-bubble interaction and jet formation by high-speed imaging and characterize the resultant flow field using particle image velocimetry (PIV). Finally, we present some applications of this technique for single cell analysis, including cell membrane poration with macromolecule uptake, localized membrane deformation determined by the displacements of attached integrin-binding beads, and intracellular calcium response from ratiometric imaging. Our results show that a fast and directional jetting flow is produced by the tandem bubble interaction, which can impose a highly-localized shear stress on the surface of a cell grown in close proximity. Furthermore, different bioeffects can be induced by altering the strength of the jetting flow by adjusting the standoff distance from the cell to the tandem bubbles. PMID:28117807

Electric potentiation of gravikinesis in Paramecium is possibly mediated by filaments.

PubMed

Machemer, H

1998-01-01

Sensitivity of Paramecium to mechanical stress including gravitational force is organized along two opposing gradients of membrane channel distribution: depolarizing Ca channels and hyperpolarizing K channels. Mechanoreceptor channels reside in the membrane of the cell soma and are activated, when the weight of the cytoplasm deforms the "lower" plasma membrane. Channel distribution is such as to generate ciliary activation which can counteract sedimentation of the cells: a reduction in downward swimming rate and an augmentation in upward swimming rate. Application of weak DC fields does not only induce the well-known cathodal orientation and swimming of Paramecium toward the cathode (galvano-taxis). We document that swimming velocity is augmented up to 175% as a function of the voltage gradient between 0.3 V/cm and 0.8 V/cm (galvanokinesis). A gradient of 0.3 V/cm was highly effective in raising the common negative gravikinesis of downward swimmers threefold. The gravikinesis of upward swimmers reversed polarity under field stimulation inducing cells to augment sedimentation effects (positive gravikinesis). Both effects of electric-field stimulation on ciliary activation are of the depolarizing type: reduction in the frequency of normally beating cilia. Analysis of the data shows that a voltage-sensitivity of gravireceptor channels would not account for the observed potentiation of negative gravikinesis. It is suggested that a previously described voltage-dependent Ca channel of the soma membrane interferes with a Ca(2+)-sensitive, peripheral filament system, which directly connects to gravireceptor channels.

Drosophila F-BAR protein Syndapin contributes to coupling the plasma membrane and contractile ring in cytokinesis.

PubMed

Takeda, Tetsuya; Robinson, Iain M; Savoian, Matthew M; Griffiths, John R; Whetton, Anthony D; McMahon, Harvey T; Glover, David M

2013-08-07

Cytokinesis is a highly ordered cellular process driven by interactions between central spindle microtubules and the actomyosin contractile ring linked to the dynamic remodelling of the plasma membrane. The mechanisms responsible for reorganizing the plasma membrane at the cell equator and its coupling to the contractile ring in cytokinesis are poorly understood. We report here that Syndapin, a protein containing an F-BAR domain required for membrane curvature, contributes to the remodelling of the plasma membrane around the contractile ring for cytokinesis. Syndapin colocalizes with phosphatidylinositol 4,5-bisphosphate (PI(4,5)P₂) at the cleavage furrow, where it directly interacts with a contractile ring component, Anillin. Accordingly, Anillin is mislocalized during cytokinesis in Syndapin mutants. Elevated or diminished expression of Syndapin leads to cytokinesis defects with abnormal cortical dynamics. The minimal segment of Syndapin, which is able to localize to the cleavage furrow and induce cytokinesis defects, is the F-BAR domain and its immediate C-terminal sequences. Phosphorylation of this region prevents this functional interaction, resulting in reduced ability of Syndapin to bind to and deform membranes. Thus, the dephosphorylated form of Syndapin mediates both remodelling of the plasma membrane and its proper coupling to the cytokinetic machinery.

Geometrically Nonlinear Shell Analysis of Wrinkled Thin-Film Membranes with Stress Concentrations

NASA Technical Reports Server (NTRS)

Tessler, Alexander; Sleight, David W.

2006-01-01

Geometrically nonlinear shell finite element analysis has recently been applied to solar-sail membrane problems in order to model the out-of-plane deformations due to structural wrinkling. Whereas certain problems lend themselves to achieving converged nonlinear solutions that compare favorably with experimental observations, solutions to tensioned membranes exhibiting high stress concentrations have been difficult to obtain even with the best nonlinear finite element codes and advanced shell element technology. In this paper, two numerical studies are presented that pave the way to improving the modeling of this class of nonlinear problems. The studies address the issues of mesh refinement and stress-concentration alleviation, and the effects of these modeling strategies on the ability to attain converged nonlinear deformations due to wrinkling. The numerical studies demonstrate that excessive mesh refinement in the regions of stress concentration may be disadvantageous to achieving wrinkled equilibrium states, causing the nonlinear solution to lock in the membrane response mode, while totally discarding the very low-energy bending response that is necessary to cause wrinkling deformation patterns.

Theoretical Analysis on Mechanical Deformation of Membrane-Based Photomask Blanks

NASA Astrophysics Data System (ADS)

Marumoto, Kenji; Aya, Sunao; Yabe, Hedeki; Okada, Tatsunori; Sumitani, Hiroaki

2012-04-01

Membrane-based photomask is used in proximity X-ray lithography including that in LIGA (Lithographie, Galvanoformung und Abformung) process, and near-field photolithography. In this article, out-of-plane deformation (OPD) and in-plane displacement (IPD) of membrane-based photomask blanks are theoretically analyzed to obtain the mask blanks with flat front surface and low stress absorber film. First, we derived the equations of OPD and IPD for the processing steps of membrane-based photomask such as film deposition, back-etching and bonding, using a theory of symmetrical bending of circular plates with a coaxial circular hole and that of deformation of cylinder under hydrostatic pressure. The validity of the equations was proved by comparing the calculation results with experimental ones. Using these equations, we investigated the relation between the geometry of the mask blanks and the distortions generally, and gave the criterion to attain the flat front surface. Moreover, the absorber stress-bias required to obtain zero-stress on finished mask blanks was also calculated and it has been found that only little stress-bias was required for adequate hole size of support plate.

Closed membrane shapes with attached BAR domains subject to external force of actin filaments.

PubMed

Mesarec, Luka; Góźdź, Wojciech; Iglič, Veronika Kralj; Kralj, Samo; Iglič, Aleš

2016-05-01

Membrane deformations induced by attached BAR superfamily domains could trigger or facilitate the growth of plasma membrane protrusions. The BAR domain family consists of BAR, F-BAR and I-BAR domains, each enforcing a different local curvature when attached to the membrane surface. Our theoretical study mainly focuses on the role of I-BAR in the membrane tubular deformations generated or stabilised by actin filaments. The influence of the area density of membrane attached BAR domains and their intrinsic curvature on the closed membrane shapes (vesicles) was investigated numerically. We derived an analytical approximative expression for the critical relative area density of BARs at which the membrane tubular protrusions on vesicles are most prominent. We have shown that the BARs with a higher intrinsic curvature induce thinner and longer cylindrical protrusions. The average orientation of the membrane attached BARs is altered when the vesicle shape is subjected to external force of growing actin rod-like structure inside a vesicle. The average orientation angle of membrane attached BARs may indicate whether the actin filaments are just stabilising the protrusion or generating it by stretching the vesicle. Copyright © 2016 Elsevier B.V. All rights reserved.

Caterpillar locomotion-inspired valveless pneumatic micropump using a single teardrop-shaped elastomeric membrane.

PubMed

So, Hongyun; Pisano, Albert P; Seo, Young Ho

2014-07-07

This paper presents a microfluidic pump operated by an asymmetrically deformed membrane, which was inspired by caterpillar locomotion. Almost all mechanical micropumps consist of two major components of fluid halting and fluid pushing parts, whereas the proposed caterpillar locomotion-inspired micropump has only a single, bilaterally symmetric membrane-like teardrop shape. A teardrop-shaped elastomeric membrane was asymmetrically deformed and then consecutively touched down to the bottom of the chamber in response to pneumatic pressure, thus achieving fluid pushing. Consecutive touchdown motions of the teardrop-shaped membrane mimicked the propagation of a caterpillar's hump during its locomotory gait. The initial touchdown motion of the teardrop-shaped membrane at the centroid worked as a valve that blocked the inlet channel, and then, the consecutive touchdown motions pushed fluid in the chamber toward the tail of the chamber connected to the outlet channel. The propagation of the touchdown motion of the teardrop-shaped membrane was investigated using computational analysis as well as experimental studies. This caterpillar locomotion-inspired micropump composed of only a single membrane can provide new opportunities for simple integration of microfluidic systems.

A Comprehensive Fluid Dynamic-Diffusion Model of Blood Microcirculation with Focus on Sickle Cell Disease

NASA Astrophysics Data System (ADS)

Le Floch, Francois; Harris, Wesley L.

2009-11-01

A novel methodology has been developed to address sickle cell disease, based on highly descriptive mathematical models for blood flow in the capillaries. Our investigations focus on the coupling between oxygen delivery and red blood cell dynamics, which is crucial to understanding sickle cell crises and is unique to this blood disease. The main part of our work is an extensive study of blood dynamics through simulations of red cells deforming within the capillary vessels, and relies on the use of a large mathematical system of equations describing oxygen transfer, blood plasma dynamics and red cell membrane mechanics. This model is expected to lead to the development of new research strategies for sickle cell disease. Our simulation model could be used not only to assess current researched remedies, but also to spur innovative research initiatives, based on our study of the physical properties coupled in sickle cell disease.

Characterization, fabrication, and analysis of soft dielectric elastomer actuators capable of complex 3D deformation

NASA Astrophysics Data System (ADS)

Lai, William

Inspired by nature, the development of soft actuators has drawn large attention to provide higher flexibility and allow adaptation to more complex environment. This thesis is focused on utilizing electroactive polymers as active materials to develop soft planar dielectric elastomer actuators capable of complex 3D deformation. The potential applications of such soft actuators are in flexible robotic arms and grippers, morphing structures and flapping wings for micro aerial vehicles. The embraces design for a freestanding actuator utilizes the constrained deformation imposed by surface stiffeners on an electroactive membrane to avert the requirement of membrane pre-stretch and the supporting frames. The proposed design increases the overall actuator flexibility and degrees-of-freedom. Actuator design, fabrication, and performance are presented for different arrangement of stiffeners. Digital images correlation technique were utilized to evaluate the in-plane finite strain components, in order to elucidate the role of the stiffeners in controlling the three dimensional deformation. It was found that a key controlling factor was the localized deformation near the stiffeners, while the rest of the membrane would follow through. A detailed finite element modeling framework was developed with a user-material subroutine, built into the ABAQUS commercial finite element package. An experimentally calibrated Neo-Hookean based material model that coupled the applied electrical field to the actuator mechanical deformation was employed. The numerical model was used to optimize different geometrical features, electrode layup and stacking sequence of actuators. It was found that by splitting the stiffeners into finer segments, the force-stroke characteristics of actuator were able to be adjusted with stiffener configuration, while keeping the overall bending stiffness. The efficacy of actuators could also be greatly improved by increasing the stiffener periodicity. The developed framework would aid in designing and optimizing the dielectric elastomer actuator configurations for 3D prescribed deformation configuration. Finally, inspired by the membrane textures of bat wings, a study of utilizing fiber reinforcement on dielectric elastomer actuators were conducted for the mechanical and the coupled electromechanical characteristics. Woven fibers were employed on the surface of actuator membrane with different pre-deformed configurations. Experimentally, actuator stiffness changes were measured for up to four orders of magnitude. The orientation of embedded fibers controlled the level and the triggered phase of stiffness changes. A trade-off between the actuator stiffness and stroke could be controlled during the fabrication stage by the fiber orientation and the prestretch level of the base elastomer membrane. A simplified model using small-strain composite laminate theory was developed and accurately predicted the composite actuator stiffness. Additionally, compliant edge stiffeners were found had to present a marked overall effect on actuator electromechanical response. The developed simplified analytical solutions using Timoshenko-bimaterial laminate solution and composite laminate theory, as well as the developed finite element framework can be utilized in addressing more complex 3D deformation patterns and their electromechanical response.

Effects of flaxseed oil on anti-oxidative system and membrane deformation of human peripheral blood erythrocytes in high glucose level.

PubMed

Yang, Wei; Fu, Juan; Yu, Miao; Huang, Qingde; Wang, Di; Xu, Jiqu; Deng, Qianchun; Yao, Ping; Huang, Fenghong; Liu, Liegang

2012-07-08

The erythrocyte membrane lesion is a serious diabetic complication. A number of studies suggested that n-3 fatty acid could reduce lipid peroxidation and elevate α- or γ-tocopherol contents in membrane of erythrocytes. However, evidence regarding the protective effects of flaxseed oil, a natural product rich in n-3 fatty acid, on lipid peroxidation, antioxidative capacity and membrane deformation of erythrocytes exposed to high glucose is limited. Human peripheral blood erythrocytes were isolated and treated with 50 mM glucose to mimic hyperglycemia in the absence or presence of three different doses of flaxseed oil (50, 100 or 200 μM) in the culture medium for 24 h. The malondialdehyde (MDA) and L-glutathione (GSH) were measured by HPLC and LC/MS respectively. The phospholipids symmetry and membrane fatty acid composition of human erythrocytes were detected by flow cytometry and gas chromatograph (GC). The morphology of human erythrocyte was illuminated by ultra scanning electron microscopy. Flaxseed oil attenuated hyperglycemia-induced increase of MDA and decrease of GSH in human erythrocytes. Human erythrocytes treated with flaxseed oil contained higher C22:5 and C22:6 than those in the 50 mM glucose control group, indicating that flaxseed oil could reduce lipid asymmetric distribution and membrane perturbation. The ultra scanning electron microscopy and flow cytometer have also indicated that flaxseed oil could protect the membrane of human erythrocytes from deformation at high glucose level. The flaxseed oil supplementation may prevent lipid peroxidation and membrane dysfunction of human erythrocytes in hyperglycemia.

Pf155/RESA protein influences the dynamic microcirculatory behavior of ring-stage Plasmodium falciparum infected red blood cells

PubMed Central

Diez-Silva, Monica; Park, YongKeun; Huang, Sha; Bow, Hansen; Mercereau-Puijalon, Odile; Deplaine, Guillaume; Lavazec, Catherine; Perrot, Sylvie; Bonnefoy, Serge; Feld, Michael S.; Han, Jongyoon; Dao, Ming; Suresh, Subra

2012-01-01

Proteins exported by Plasmodium falciparum to the red blood cell (RBC) membrane modify the structural properties of the parasitized RBC (Pf-RBC). Although quasi-static single cell assays show reduced ring-stage Pf-RBCs deformability, the parameters influencing their microcirculatory behavior remain unexplored. Here, we study the dynamic properties of ring-stage Pf-RBCs and the role of the parasite protein Pf155/Ring-Infected Erythrocyte Surface Antigen (RESA). Diffraction phase microscopy revealed RESA-driven decreased Pf-RBCs membrane fluctuations. Microfluidic experiments showed a RESA-dependent reduction in the Pf-RBCs transit velocity, which was potentiated at febrile temperature. In a microspheres filtration system, incubation at febrile temperature impaired traversal of RESA-expressing Pf-RBCs. These results show that RESA influences ring-stage Pf-RBCs microcirculation, an effect that is fever-enhanced. This is the first identification of a parasite factor influencing the dynamic circulation of young asexual Pf-RBCs in physiologically relevant conditions, offering novel possibilities for interventions to reduce parasite survival and pathogenesis in its human host. PMID:22937223

Pf155/RESA protein influences the dynamic microcirculatory behavior of ring-stage Plasmodium falciparum infected red blood cells

NASA Astrophysics Data System (ADS)

Diez-Silva, Monica; Park, Yongkeun; Huang, Sha; Bow, Hansen; Mercereau-Puijalon, Odile; Deplaine, Guillaume; Lavazec, Catherine; Perrot, Sylvie; Bonnefoy, Serge; Feld, Michael S.; Han, Jongyoon; Dao, Ming; Suresh, Subra

2012-08-01

Proteins exported by Plasmodium falciparum to the red blood cell (RBC) membrane modify the structural properties of the parasitized RBC (Pf-RBC). Although quasi-static single cell assays show reduced ring-stage Pf-RBCs deformability, the parameters influencing their microcirculatory behavior remain unexplored. Here, we study the dynamic properties of ring-stage Pf-RBCs and the role of the parasite protein Pf155/Ring-Infected Erythrocyte Surface Antigen (RESA). Diffraction phase microscopy revealed RESA-driven decreased Pf-RBCs membrane fluctuations. Microfluidic experiments showed a RESA-dependent reduction in the Pf-RBCs transit velocity, which was potentiated at febrile temperature. In a microspheres filtration system, incubation at febrile temperature impaired traversal of RESA-expressing Pf-RBCs. These results show that RESA influences ring-stage Pf-RBCs microcirculation, an effect that is fever-enhanced. This is the first identification of a parasite factor influencing the dynamic circulation of young asexual Pf-RBCs in physiologically relevant conditions, offering novel possibilities for interventions to reduce parasite survival and pathogenesis in its human host.

Asymmetric Mbc, active Rac1 and F-actin foci in the fusion-competent myoblasts during myoblast fusion in Drosophila

PubMed Central

Haralalka, Shruti; Shelton, Claude; Cartwright, Heather N.; Katzfey, Erin; Janzen, Evan; Abmayr, Susan M.

2011-01-01

Myoblast fusion is an intricate process that is initiated by cell recognition and adhesion, and culminates in cell membrane breakdown and formation of multinucleate syncytia. In the Drosophila embryo, this process occurs asymmetrically between founder cells that pattern the musculature and fusion-competent myoblasts (FCMs) that account for the bulk of the myoblasts. The present studies clarify and amplify current models of myoblast fusion in several important ways. We demonstrate that the non-conventional guanine nucleotide exchange factor (GEF) Mbc plays a fundamental role in the FCMs, where it functions to activate Rac1, but is not required in the founder cells for fusion. Mbc, active Rac1 and F-actin foci are highly enriched in the FCMs, where they localize to the Sns:Kirre junction. Furthermore, Mbc is crucial for the integrity of the F-actin foci and the FCM cytoskeleton, presumably via its activation of Rac1 in these cells. Finally, the local asymmetric distribution of these proteins at adhesion sites is reminiscent of invasive podosomes and, consistent with this model, they are enriched at sites of membrane deformation, where the FCM protrudes into the founder cell/myotube. These data are consistent with models promoting actin polymerization as the driving force for myoblast fusion. PMID:21389053

FURTHER STUDY OF SOMA, DENDRITE, AND AXON EXCITATION IN SINGLE NEURONS

PubMed Central

Eyzaguirre, Carlos; Kuffler, Stephen W.

1955-01-01

The present investigation continues a previous study in which the soma-dendrite system of sensory neurons was excited by stretch deformation of the peripheral dendrite portions. Recording was done with intracellular leads which were inserted into the cell soma while the neuron was activated orthodromically or antidromically. The analysis was also extended to axon conduction. Crayfish, Procambarus alleni (Faxon) and Orconectes virilis (Hagen), were used. 1. The size and time course of action potentials recorded from the soma-dendrite complex vary greatly with the level of the cell's membrane potential. The latter can be changed over a wide range by stretch deformation which sets up a "generator potential" in the distal portions of the dendrites. If a cell is at its resting unstretched equilibrium potential, antidromic stimulation through the axon causes an impulse which normally overshoots the resting potential and decays into an afternegativity of 15 to 20 msec. duration. The postspike negativity is not followed by an appreciable hyperpolarization (positive) phase. If the membrane potential is reduced to a new steady level a postspike positivity appears and increases linearly over a depolarization range of 12 to 20 mv. in various cells. At those levels the firing threshold of the cell for orthodromic discharges is generally reached. 2. The safety factor for conduction between axon and cell soma is reduced under three unrelated conditions, (a) During the recovery period (2 to 3 msec.) immediately following an impulse which has conducted fully over the cell soma, a second impulse may be delayed, may invade the soma partially, or may be blocked completely. (b) If progressive depolarization is produced by stretch, it leads to a reduction of impulse height and eventually to complete block of antidromic soma invasion, resembling cathodal block, (c) In some cells, when the normal membrane potential is within several millivolts of the relaxed resting state, an antidromic impulse may be blocked and may set up within the soma a local potential only. The local potential can sum with a second one or it may sum with potential changes set up in the dendrites, leading to complete invasion of the soma. Such antidromic invasion block can always be relieved by appropriate stretch which shifts the membrane potential out of the "blocking range" nearer to the soma firing level. During the afterpositivity of an impulse in a stretched cell the membrane potential may fall below or near the blocking range. During that period another impulse may be delayed or blocked. 3. Information regarding activity and conduction in dendrites has been obtained indirectly, mainly by analyzing the generator action under various conditions of stretch. The following conclusions have been reached: The large dendrite branches have similar properties to the cell body from which they arise and carry the same kind of impulses. In the finer distal filaments of even lightly depolarized dendrites, however, no axon type all-or-none conduction occurs since the generator potential persists to a varying degree during antidromic invasion of the cell. With the membrane potential at its resting level the dendrite terminals contribute to the prolonged impulse afternegativity of the soma. 4. Action potentials in impaled axons and in cell bodies have been compared. It is thought that normally the over-all duration of axon impulses is shorter. Local activity during reduction of the safety margin for conduction was studied. 5. An analysis was made of high frequency grouped discharges which occasionally arise in cells. They differ in many essential aspects from the regular discharges set up by the generator action. It is proposed that grouped discharges occur only when invasion of dendrites is not synchronous, due to a delay in excitation spread between soma and dendrites. Each impulse in a group is assumed to be caused by an impulse in at least one of the large dendrite branches. Depolarization of dendrites abolishes the grouped activity by facilitating invasion of the large dendrite branches. PMID:13252238

Nanomechanical properties of phospholipid microbubbles.

PubMed

Buchner Santos, Evelyn; Morris, Julia K; Glynos, Emmanouil; Sboros, Vassilis; Koutsos, Vasileios

2012-04-03

This study uses atomic force microscopy (AFM) force-deformation (F-Δ) curves to investigate for the first time the Young's modulus of a phospholipid microbubble (MB) ultrasound contrast agent. The stiffness of the MBs was calculated from the gradient of the F-Δ curves, and the Young's modulus of the MB shell was calculated by employing two different mechanical models based on the Reissner and elastic membrane theories. We found that the relatively soft phospholipid-based MBs behave inherently differently to stiffer, polymer-based MBs [Glynos, E.; Koutsos, V.; McDicken, W. N.; Moran, C. M.; Pye, S. D.; Ross, J. A.; Sboros, V. Langmuir2009, 25 (13), 7514-7522] and that elastic membrane theory is the most appropriate of the models tested for evaluating the Young's modulus of the phospholipid shell, agreeing with values available for living cell membranes, supported lipid bilayers, and synthetic phospholipid vesicles. Furthermore, we show that AFM F-Δ curves in combination with a suitable mechanical model can assess the shell properties of phospholipid MBs. The "effective" Young's modulus of the whole bubble was also calculated by analysis using Hertz theory. This analysis yielded values which are in agreement with results from studies which used Hertz theory to analyze similar systems such as cells.

Computational model of deformable lenses actuated by dielectric elastomers

NASA Astrophysics Data System (ADS)

Lu, Tongqing; Cai, Shengqiang; Wang, Huiming; Suo, Zhigang

2013-09-01

A recent design of deformable lens mimics the human eye, adjusting its focal length in response to muscle-like actuation. The artificial muscle is a membrane of a dielectric elastomer subject to a voltage. Here, we calculate the coupled and inhomogeneous deformation of the lens and the dielectric elastomer actuator by formulating a nonlinear boundary-value problem. We characterize the strain-stiffening elastomer with the Gent model and describe the voltage-induced deformation using the model of ideal dielectric elastomer. The computational predictions agree well with experimental data. We use the model to explore the space of parameters, including the prestretch of the membrane, the volume of the liquid in the lens, and the size of the dielectric elastomer actuator relative to the lens. We examine how various modes of failure limit the minimum radius of curvature.

Interaction measurement of particles bound to a lipid membrane

NASA Astrophysics Data System (ADS)

Sarfati, Raphael; Dufresne, Eric

2015-03-01

The local shape and dynamics of the plasma membrane play important roles in many cellular processes. Local membrane deformations are often mediated by the adsorption of proteins (notably from the BAR family), and their subsequent self-assembly. The emerging hypothesis is that self-assembly arises from long-range interactions of individual proteins through the membrane's deformation field. We study these interactions in a model system of micron-sized colloidal particles adsorbed onto a lipid bilayer. We use fluorescent microscopy, optical tweezers and particle tracking to measure dissipative and conservative forces as a function of the separation between the particles. We find that particles are driven together with forces of order 100 fN and remain bound in a potential well with a stiffness of order 100 fN/micron.

Force-Mediating Magnetic Nanoparticles to Engineer Neuronal Cell Function

PubMed Central

Gahl, Trevor J.; Kunze, Anja

2018-01-01

Cellular processes like membrane deformation, cell migration, and transport of organelles are sensitive to mechanical forces. Technically, these cellular processes can be manipulated through operating forces at a spatial precision in the range of nanometers up to a few micrometers through chaperoning force-mediating nanoparticles in electrical, magnetic, or optical field gradients. But which force-mediating tool is more suitable to manipulate cell migration, and which, to manipulate cell signaling? We review here the differences in forces sensation to control and engineer cellular processes inside and outside the cell, with a special focus on neuronal cells. In addition, we discuss technical details and limitations of different force-mediating approaches and highlight recent advancements of nanomagnetics in cell organization, communication, signaling, and intracellular trafficking. Finally, we give suggestions about how force-mediating nanoparticles can be used to our advantage in next-generation neurotherapeutic devices. PMID:29867315

Force-Mediating Magnetic Nanoparticles to Engineer Neuronal Cell Function.

PubMed

Gahl, Trevor J; Kunze, Anja

2018-01-01

Cellular processes like membrane deformation, cell migration, and transport of organelles are sensitive to mechanical forces. Technically, these cellular processes can be manipulated through operating forces at a spatial precision in the range of nanometers up to a few micrometers through chaperoning force-mediating nanoparticles in electrical, magnetic, or optical field gradients. But which force-mediating tool is more suitable to manipulate cell migration, and which, to manipulate cell signaling? We review here the differences in forces sensation to control and engineer cellular processes inside and outside the cell, with a special focus on neuronal cells. In addition, we discuss technical details and limitations of different force-mediating approaches and highlight recent advancements of nanomagnetics in cell organization, communication, signaling, and intracellular trafficking. Finally, we give suggestions about how force-mediating nanoparticles can be used to our advantage in next-generation neurotherapeutic devices.

A Continuum Method for Determining Membrane Protein Insertion Energies and the Problem of Charged Residues

PubMed Central

Choe, Seungho; Hecht, Karen A.; Grabe, Michael

2008-01-01

Continuum electrostatic approaches have been extremely successful at describing the charged nature of soluble proteins and how they interact with binding partners. However, it is unclear whether continuum methods can be used to quantitatively understand the energetics of membrane protein insertion and stability. Recent translation experiments suggest that the energy required to insert charged peptides into membranes is much smaller than predicted by present continuum theories. Atomistic simulations have pointed to bilayer inhomogeneity and membrane deformation around buried charged groups as two critical features that are neglected in simpler models. Here, we develop a fully continuum method that circumvents both of these shortcomings by using elasticity theory to determine the shape of the deformed membrane and then subsequently uses this shape to carry out continuum electrostatics calculations. Our method does an excellent job of quantitatively matching results from detailed molecular dynamics simulations at a tiny fraction of the computational cost. We expect that this method will be ideal for studying large membrane protein complexes. PMID:18474636

Soft x-ray spectroscopy of high pressure liquid.

PubMed

Qiao, Ruimin; Xia, Yujian; Feng, Xuefei; Macdougall, James; Pepper, John; Armitage, Kevin; Borsos, Jason; Knauss, Kevin G; Lee, Namhey; Allézy, Arnaud; Gilbert, Benjamin; MacDowell, Alastair A; Liu, Yi-Sheng; Glans, Per-Anders; Sun, Xuhui; Chao, Weilun; Guo, Jinghua

2018-01-01

We describe a new experimental technique that allows for soft x-ray spectroscopy studies (∼100-1000 eV) of high pressure liquid (∼100 bars). We achieve this through a liquid cell with a 100 nm-thick Si 3 N 4 membrane window, which is sandwiched by two identical O-rings for vacuum sealing. The thin Si 3 N 4 membrane allows soft x-rays to penetrate, while separating the high-pressure liquid under investigation from the vacuum required for soft x-ray transmission and detection. The burst pressure of the Si 3 N 4 membrane increases with decreasing size and more specifically is inversely proportional to the side length of the square window. It also increases proportionally with the membrane thickness. Pressures > 60 bars could be achieved for 100 nm-thick square Si 3 N 4 windows that are smaller than 65 μm. However, above a certain pressure, the failure of the Si wafer becomes the limiting factor. The failure pressure of the Si wafer is sensitive to the wafer thickness. Moreover, the deformation of the Si 3 N 4 membrane is quantified using vertical scanning interferometry. As an example of the performance of the high-pressure liquid cell optimized for total-fluorescence detected soft x-ray absorption spectroscopy (sXAS), the sXAS spectra at the Ca L edge (∼350 eV) of a CaCl 2 aqueous solution are collected under different pressures up to 41 bars.

Soft x-ray spectroscopy of high pressure liquid

DOE PAGES

Qiao, Ruimin; Xia, Yujian; Feng, Xuefei; ...

2018-01-01

Here, we describe a new experimental technique that allows for soft x-ray spectroscopy studies (~100-1000 eV) of high pressure liquid (~100 bars). We achieve this through a liquid cell with a 100 nm-thick Si 3N 4 membrane window, which is sandwiched by two identical O-rings for vacuum sealing. The thin Si 3N 4 membrane allows soft x-rays to penetrate, while separating the high-pressure liquid under investigation from the vacuum required for soft x-ray transmission and detection. The burst pressure of the Si 3N 4 membrane increases with decreasing size and more specifically is inversely proportional to the side length ofmore » the square window. It also increases proportionally with the membrane thickness. Pressures > 60 bars could be achieved for 100 nm-thick square Si 3N 4 windows that are smaller than 65 μm. However, above a certain pressure, the failure of the Si wafer becomes the limiting factor. The failure pressure of the Si wafer is sensitive to the wafer thickness. Moreover, the deformation of the Si 3N 4 membrane is quantified using vertical scanning interferometry. As an example of the performance of the high-pressure liquid cell optimized for total-fluorescence detected soft x-ray absorption spectroscopy (sXAS), the sXAS spectra at the Ca L edge (~350 eV) of a CaCl 2 aqueous solution are collected under different pressures up to 41 bars.« less

Soft x-ray spectroscopy of high pressure liquid

DOE Office of Scientific and Technical Information (OSTI.GOV)

Qiao, Ruimin; Xia, Yujian; Feng, Xuefei

Here, we describe a new experimental technique that allows for soft x-ray spectroscopy studies (~100-1000 eV) of high pressure liquid (~100 bars). We achieve this through a liquid cell with a 100 nm-thick Si 3N 4 membrane window, which is sandwiched by two identical O-rings for vacuum sealing. The thin Si 3N 4 membrane allows soft x-rays to penetrate, while separating the high-pressure liquid under investigation from the vacuum required for soft x-ray transmission and detection. The burst pressure of the Si 3N 4 membrane increases with decreasing size and more specifically is inversely proportional to the side length ofmore » the square window. It also increases proportionally with the membrane thickness. Pressures > 60 bars could be achieved for 100 nm-thick square Si 3N 4 windows that are smaller than 65 μm. However, above a certain pressure, the failure of the Si wafer becomes the limiting factor. The failure pressure of the Si wafer is sensitive to the wafer thickness. Moreover, the deformation of the Si 3N 4 membrane is quantified using vertical scanning interferometry. As an example of the performance of the high-pressure liquid cell optimized for total-fluorescence detected soft x-ray absorption spectroscopy (sXAS), the sXAS spectra at the Ca L edge (~350 eV) of a CaCl 2 aqueous solution are collected under different pressures up to 41 bars.« less

Acoustic investigation of the aperture dynamics of an elastic membrane closing an overpressurized cylindrical cavity

NASA Astrophysics Data System (ADS)

Sánchez, Claudia; Vidal, Valérie; Melo, Francisco

2015-08-01

We report an experimental study of the acoustic signal produced by the rupture of an elastic membrane that initially closes a cylindrical overpressurized cavity. This configuration has been recently used as an experimental model system for the investigation of the acoustic emission from the bursting of elongated gas bubbles rising in a conduit. Here, we investigate the effect of the membrane rupture dynamics on the acoustic signal produced by the pressure release by changing the initial tension of the membrane. The initial overpressure in the cavity is fixed at a value such that the system remains in the linear acoustic regime. For large initial membrane deformation, the rupture time τ rup is small compared to the wave propagation time in the cavity and the pressure wave inside the conduit can be fully captured by the linear theory. For low membrane tension, a hole is pierced in the membrane but its rupture does not occur. For intermediate deformation, finally, the rupture progresses in two steps: first the membrane opens slowly; then, after reaching a critical size, the rupture accelerates. A transversal wave is excited along the membrane surface. The characteristic signature of each opening dynamics on the acoustic emission is described.

Vibration mode analysis of the proton exchange membrane fuel cell stack

NASA Astrophysics Data System (ADS)

Liu, B.; Liu, L. F.; Wei, M. Y.; Wu, C. W.

2016-11-01

Proton exchange membrane fuel cell (PEMFC) stacks usually undergo vibration during packing, transportation, and serving time, in particular for those used in the automobiles or portable equipment. To study the stack vibration response, based on finite element method (FEM), a mode analysis is carried out in the present paper. Using this method, we can distinguish the local vibration from the stack global modes, predict the vibration responses, such as deformed shape and direction, and discuss the effects of the clamping configuration and the clamping force magnitude on vibration modes. It is found that when the total clamping force remains the same, increasing the bolt number can strengthen the stack resistance to vibration in the clamping direction, but cannot obviously strengthen stack resistance to vibration in the translations perpendicular to clamping direction and the three axis rotations. Increasing the total clamping force can increase both of the stack global mode and the bolt local mode frequencies, but will decrease the gasket local mode frequency.

Combinatorial approaches to evaluate nanodiamond uptake and induced cellular fate

NASA Astrophysics Data System (ADS)

Eldawud, Reem; Reitzig, Manuela; Opitz, Jörg; Rojansakul, Yon; Jiang, Wenjuan; Nangia, Shikha; Zoica Dinu, Cerasela

2016-02-01

Nanodiamonds (NDs) are an emerging class of engineered nanomaterials that hold great promise for the next generation of bionanotechnological products to be used for drug and gene delivery, or for bio-imaging and biosensing. Previous studies have shown that upon their cellular uptake, NDs exhibit high biocompatibility in various in vitro and in vivo set-ups. Herein we hypothesized that the increased NDs biocompatibility is a result of minimum membrane perturbations and their reduced ability to induce disruption or damage during cellular translocation. Using multi-scale combinatorial approaches that simulate ND-membrane interactions, we correlated NDs real-time cellular uptake and kinetics with the ND-induced membrane fluctuations to derive energy requirements for the uptake to occur. Our discrete and real-time analyses showed that the majority of NDs internalization occurs within 2 h of cellular exposure, however, with no effects on cellular viability, proliferation or cellular behavior. Furthermore, our simulation analyses using coarse-grained models identified key changes in the energy profile, membrane deformation and recovery time, all functions of the average ND or ND-based agglomerate size. Understanding the mechanisms responsible for ND-cell membrane interactions could possibly advance their implementation in various biomedical applications.

Combinatorial approaches to evaluate nanodiamond uptake and induced cellular fate

PubMed Central

Eldawud, Reem; Reitzig, Manuela; Opitz, Jörg; Rojansakul, Yon; Jiang, Wenjuan; Nangia, Shikha; Dinu, Cerasela Zoica

2016-01-01

Nanodiamonds (NDs) are an emerging class of engineered nanomaterials that hold great promise for the next generation of bionanotechnological products to be used for drug and gene delivery, or for bio-imaging and biosensing. Previous studies have shown that upon their cellular uptake, NDs exhibit high biocompatibility in various in vitro and in vivo set-ups. Herein we hypothesized that the increased NDs biocompatibility is a result of minimum membrane perturbations and their reduced ability to induce disruption or damage during cellular translocation. Using multi-scale combinatorial approaches that simulate ND-membrane interactions, we correlated NDs real-time cellular uptake and kinetics with the ND-induced membrane fluctuations to derive energy requirements for the uptake to occur. Our discrete and real-time analyses showed that the majority of NDs internalization occurs within 2 h of cellular exposure, however, with no effects on cellular viability, proliferation or cellular behavior. Furthermore, our simulation analyses using coarse-grained models identified key changes in the energy profile, membrane deformation and recovery time, all functions of the average ND or ND-based agglomerate size. Understanding the mechanisms responsible for ND-cell membrane interactions could possibly advance their implementation in various biomedical applications. PMID:26820775

In vivo influence of extract from Aronia melanocarpa on the erythrocyte membranes in patients with hypercholesterolemia

PubMed Central

Duchnowicz, Piotr; Nowicka, Agnieszka; Koter-Michalak, Maria; Broncel, Marlena

2012-01-01

Summary Background Hypercholesterolemia increases cholesterol concentration in erythrocyte membranes, which results in decrease of membrane fluidity and decreases the deformability of red blood cells. The fruits of Arona melanocarpa contains many of polyphenols and other compounds that have beneficial health effects. Material/Methods The aim of the study was to estimate the influence of 2-month supplementation of extract from Aronia melanocarpa (100 mg Aronox, three times per day) on cholesterol concentration, lipid peroxidation, membrane fluidity, level of thiol groups and activity of ATPase in erythrocytes from patients with hypercholesterolemia. The study involved 25 patients with hypercholesterolemia without pharmacological treatment and 20 healthy individuals as a control group. Blood samples were collected before, and after 1 and 2 months of Aronia administration. Results The 2-month Aronia supplementation resulted in a decrease of cholesterol concentration (by 22%) and a decrease of lipid peroxidation (by 40%), and an increase of membrane fluidity. No statistically significant increase of the concentration of thiol groups and of ATPase activity were observed. Conclusions Our study shows that supplementation of extract from Aronia melanocarpa has a beneficial effect on rheological properties of erythrocytes. PMID:22936193

Versatile Miniature Tunable Liquid Lenses Using Transparent Graphene Electrodes.

PubMed

Shahini, Ali; Xia, Jinjun; Zhou, Zhixian; Zhao, Yang; Cheng, Mark Ming-Cheng

2016-02-16

This paper presents, for the first time, versatile and low-cost miniature liquid lenses with graphene as electrodes. Tunable focal length is achieved by changing the droplet curvature using electrowetting on dielectric (EWOD). Ionic liquid and KCl solution are utilized as lens liquid on the top of a flexible Teflon-coated PDMS/parylene membrane. Transparent and flexible, graphene allows transmission of visible light as well as large deformation of the polymer membrane to achieve requirements for different lens designs and to increase the field of view without damaging of electrodes. The tunable range for the focal length is between 3 and 7 mm for a droplet with a volume of 3 μL. The visualization of bone marrow dendritic cells is demonstrated by the liquid lens system with a high resolution (456 lp/mm).

Early alterations of red blood cell rheology in critically ill patients.

PubMed

Reggiori, Giulia; Occhipinti, Giovanna; De Gasperi, Andrea; Vincent, Jean-Louis; Piagnerelli, Michael

2009-12-01

To investigate red blood cell rheology in a large intensive care unit population on admission, and to assess the possible influence of comorbidities on the rheology. : Prospective study. Medico-surgical intensive care unit with 31 beds. All intensive care unit admissions during a 5-month period and 20 healthy volunteers. Blood sampling. A total of 196 intensive care patients (160 without and 36 with sepsis) and 20 healthy volunteers were studied. Red blood cell rheology (deformability and aggregation) was assessed ex vivo using the laser-assisted optical rotational cell analyzer (LORCA; Mechatronics Instruments BV, AN Zwaag, Netherlands) within the first 24 hrs after intensive care unit admission. Red blood cell deformability was determined by the elongation index in relation to the shear stress (0.3 to 50 Pa) applied on the red blood cell membrane surface. Aggregation was assessed by the aggregation index. Septic patients were more likely to have anemia, coagulation abnormalities, and comorbidities than were nonseptic patients. Red blood cell deformability was significantly altered in septic compared to nonseptic patients and volunteers for the majority of shear stress rates studied. The aggregation index was greater in septic patients than in volunteers (67.9% [54.7-73.5] vs. 61.8% [58.2-68.4]; p < .05). Only sepsis and hematologic disease influenced the elongation index (both p < .01). Other comorbidities, like cancer, diabetes mellitus, cirrhosis, and terminal renal failure, had no effect on the elongation index. Aggregation index was related to the degree of organ failure (Sequential Organ Failure Assessment score), the red blood cell count, and fibrinogen concentrations. Early alterations of red blood cell rheology are common in intensive care unit patients, especially in those with sepsis. Comorbidities (other than hematologic diseases) do not significantly influence these abnormalities. These alterations could contribute to the microcirculatory alterations observed in critically ill patients.

Loading system mechanism for dielectric elastomer generators with equi-biaxial state of deformation

NASA Astrophysics Data System (ADS)

Fontana, M.; Moretti, G.; Lenzo, B.; Vertechy, R.

2014-03-01

Dielectric Elastomer Generators (DEGs) are devices that employ a cyclically variable membrane capacitor to produce electricity from oscillating sources of mechanical energy. Capacitance variation is obtained thanks to the use of dielectric and conductive layers that can undergo different states of deformation including: uniform or non-uniform and uni- or multi-axial stretching. Among them, uniform equi-biaxial stretching is reputed as being the most effective state of deformation that maximizes the amount of energy that can be extracted in a cycle by a unit volume of Dielectric Elastomer (DE) material. This paper presents a DEG concept, with linear input motion and tunable impedance, that is based on a mechanical loading system for inducing uniform equi-biaxial states of deformation. The presented system employs two circular DE membrane capacitors that are arranged in an agonist-antagonist configuration. An analytical model of the overall system is developed and used to find the optimal design parameters that make it possible to tune the elastic response of the generator over the range of motion of interest. An apparatus is developed for the equi-biaxial testing of DE membranes and used for the experimental verification of the employed numerical models.

Optical micromanipulation of nanoparticles and cells inside living zebrafish.

PubMed

Johansen, Patrick Lie; Fenaroli, Federico; Evensen, Lasse; Griffiths, Gareth; Koster, Gerbrand

2016-03-21

Regulation of biological processes is often based on physical interactions between cells and their microenvironment. To unravel how and where interactions occur, micromanipulation methods can be used that offer high-precision control over the duration, position and magnitude of interactions. However, lacking an in vivo system, micromanipulation has generally been done with cells in vitro, which may not reflect the complex in vivo situation inside multicellular organisms. Here using optical tweezers we demonstrate micromanipulation throughout the transparent zebrafish embryo. We show that different cells, as well as injected nanoparticles and bacteria can be trapped and that adhesion properties and membrane deformation of endothelium and macrophages can be analysed. This non-invasive micromanipulation inside a whole-organism gives direct insights into cell interactions that are not accessible using existing approaches. Potential applications include screening of nanoparticle-cell interactions for cancer therapy or tissue invasion studies in cancer and infection biology.

Effects of flaxseed oil on anti-oxidative system and membrane deformation of human peripheral blood erythrocytes in high glucose level

PubMed Central

2012-01-01

Background The erythrocyte membrane lesion is a serious diabetic complication. A number of studies suggested that n-3 fatty acid could reduce lipid peroxidation and elevate α- or γ-tocopherol contents in membrane of erythrocytes. However, evidence regarding the protective effects of flaxseed oil, a natural product rich in n-3 fatty acid, on lipid peroxidation, antioxidative capacity and membrane deformation of erythrocytes exposed to high glucose is limited. Methods Human peripheral blood erythrocytes were isolated and treated with 50 mM glucose to mimic hyperglycemia in the absence or presence of three different doses of flaxseed oil (50, 100 or 200 μM) in the culture medium for 24 h. The malondialdehyde (MDA) and L-glutathione (GSH) were measured by HPLC and LC/MS respectively. The phospholipids symmetry and membrane fatty acid composition of human erythrocytes were detected by flow cytometry and gas chromatograph (GC). The morphology of human erythrocyte was illuminated by ultra scanning electron microscopy. Results Flaxseed oil attenuated hyperglycemia-induced increase of MDA and decrease of GSH in human erythrocytes. Human erythrocytes treated with flaxseed oil contained higher C22:5 and C22:6 than those in the 50 mM glucose control group, indicating that flaxseed oil could reduce lipid asymmetric distribution and membrane perturbation. The ultra scanning electron microscopy and flow cytometer have also indicated that flaxseed oil could protect the membrane of human erythrocytes from deformation at high glucose level. Conclusion The flaxseed oil supplementation may prevent lipid peroxidation and membrane dysfunction of human erythrocytes in hyperglycemia. PMID:22768971

Effect of sterol esters on lipid composition and antioxidant status of erythrocyte membrane of hypercholesterolemic rats.

PubMed

Sengupta, Avery; Ghosh, Mahua

2014-01-01

Hypercholesterolemia is a major cause of coronary heart disease. Erythrocyte membrane is affected during hypercholesterolemia. The effect of EPA-DHA rich sterol ester and ALA rich sterol ester on erythrocyte membrane composition, osmotic fragility in normal and hypercholesterolemic rats and changes in antioxidant status of erythrocyte membrane were studied. Erythrocyte membrane composition, osmotic fragility of the membrane and antioxidant enzyme activities was analyzed. Osmotic fragility data suggested that the erythrocyte membrane of hypercholesterolemia was relatively more fragile than that of the normal rats' membrane which could be reversed with the addition of sterol esters in the diet. The increased plasma cholesterol in hypercholesterolemic rats could also be lowered by the sterol ester administration. There was also marked changes in the antioxidant enzyme activities of the erythrocyte membrane. Antioxidant enzyme levels decreased in the membrane of the hypercholesterolemic subjects were increased with the treatment of the sterol esters. The antioxidative activity of ALA rich sterol ester was better in comparison to EPA-DHA rich sterol ester. In conclusion, rat erythrocytes appear to be deformed and became more fragile in cholesterol rich blood. This deformity and fragility was partially reversed by sterol esters by virtue of their ability to lower the extent of hypercholesterolemia.

Micropore Geometry Manipulation by Macroscopic Deformation Based on Shape Memory Effect in Porous PLLA Membrane and its Enhanced Separation Performance.

PubMed

Zhao, Jingxin; Yang, Qiucheng; Wang, Tao; Wang, Lian; You, Jichun; Li, Yongjin

2017-12-20

An effective strategy to tailor the microporous structures has been developed based on the shape memory effect in porous poly(l-lactic acid) membranes in which tiny crystals and amorphous matrix play the roles of shape-fixed phase and reversible-phase, respectively. Our results indicate that not only PLLA membranes but micropores exhibit shape memory properties. The proportional deformations on two scales have been achieved by uniaxial or biaxial tension, providing a facile way to manipulate continuously the size and the orientation degree of pores on microscale. The enhanced separation performance has been validated by taking polystyrene colloids with varying diameters as an example.

Contact behavior modelling and its size effect on proton exchange membrane fuel cell

NASA Astrophysics Data System (ADS)

Qiu, Diankai; Peng, Linfa; Yi, Peiyun; Lai, Xinmin; Janßen, Holger; Lehnert, Werner

2017-10-01

Contact behavior between the gas diffusion layer (GDL) and bipolar plate (BPP) is of significant importance for proton exchange membrane fuel cells. Most current studies on contact behavior utilize experiments and finite element modelling and focus on fuel cells with graphite BPPs, which lead to high costs and huge computational requirements. The objective of this work is to build a more effective analytical method for contact behavior in fuel cells and investigate the size effect resulting from configuration alteration of channel and rib (channel/rib). Firstly, a mathematical description of channel/rib geometry is outlined in accordance with the fabrication of metallic BPP. Based on the interface deformation characteristic and Winkler surface model, contact pressure between BPP and GDL is then calculated to predict contact resistance and GDL porosity as evaluative parameters of contact behavior. Then, experiments on BPP fabrication and contact resistance measurement are conducted to validate the model. The measured results demonstrate an obvious dependence on channel/rib size. Feasibility of the model used in graphite fuel cells is also discussed. Finally, size factor is proposed for evaluating the rule of size effect. Significant increase occurs in contact resistance and porosity for higher size factor, in which channel/rib width decrease.

Property development for biaxial drawing of ethylene-tetrafluoroehtylene copolymer films and resultant fractural behavior analyzed by in situ X-ray measurements.

PubMed

Uehara, Hiroki; Ono, Yasunori; Kakiage, Masaki; Sakamura, Takumi; Masunaga, Hiroyasu; Yukawa, Yasumasa; Higuchi, Yoshiaki; Kamiya, Hiroki; Yamanobe, Takeshi

2015-03-19

The property development of the ethylene-tetrafluoroethylene copolymer (ETFE) membrane induced by simultaneous biaxial drawing was investigated. Commonly, tensile strength can be increased by drawing; conversely, tear resistance decreases. In this study, the balance between tensile strength and tear resistance for the resultant ETFE membrane was optimized achieved by a combination of lamination of low molecular weight (LMW) and high molecular weight (HMW) layers and subsequent biaxial drawing. The structural factor determining tear resistance of these biaxially drawn membranes was determined based on in situ small-angle X-ray scattering (SAXS) measurement during tensile deformation simulating tearing tests. Lozenge shaped scattering, which indicated inclined lamellae, was observed during such tensile deformation of the resultant membranes. Remarkably, this inclined lamellar structure was observed for the pure LMW membrane; however, it also appeared at the interface between LMW and HMW layers within biaxially drawn membranes. For the membrane exhibiting the highest tearing strength, the fraction of such inclined lamella increased up to the critical strain corresponding to the actual sample breaking. These results confirm that the inclined lamellar structure absorbed strain during membrane tearing.

Continuum Approaches to Understanding Ion and Peptide Interactions with the Membrane

PubMed Central

Latorraca, Naomi R.; Callenberg, Keith M.; Boyle, Jon P.; Grabe, Michael

2014-01-01

Experimental and computational studies have shown that cellular membranes deform to stabilize the inclusion of transmembrane (TM) proteins harboring charge. Recent analysis suggests that membrane bending helps to expose charged and polar residues to the aqueous environment and polar head groups. We previously used elasticity theory to identify membrane distortions that minimize the insertion of charged TM peptides into the membrane. Here, we extend our work by showing that it also provides a novel, computationally efficient method for exploring the energetics of ion and small peptide penetration into membranes. First, we show that the continuum method accurately reproduces energy profiles and membrane shapes generated from molecular simulations of bare ion permeation at a fraction of the computational cost. Next, we demonstrate that the dependence of the ion insertion energy on the membrane thickness arises primarily from the elastic properties of the membrane. Moreover, the continuum model readily provides a free energy decomposition into components not easily determined from molecular dynamics. Finally, we show that the energetics of membrane deformation strongly depend on membrane patch size both for ions and peptides. This dependence is particularly strong for peptides based on simulations of a known amphipathic, membrane binding peptide from the human pathogen Toxoplasma gondii. In total, we address shortcomings and advantages that arise from using a variety of computational methods in distinct biological contexts. PMID:24652510

Development of Sample Verification System for Sample Return Missions

NASA Technical Reports Server (NTRS)

Toda, Risaku; McKinney, Colin; Jackson, Shannon P.; Mojarradi, Mohammad; Trebi-Ollennu, Ashitey; Manohara, Harish

2011-01-01

This paper describes the development of a proof of-concept sample verification system (SVS) for in-situ mass measurement of planetary rock and soil sample in future robotic sample return missions. Our proof-of-concept SVS device contains a 10 cm diameter pressure sensitive elastic membrane placed at the bottom of a sample canister. The membrane deforms under the weight of accumulating planetary sample. The membrane is positioned in proximity to an opposing substrate with a narrow gap. The deformation of the membrane makes the gap to be narrower, resulting in increased capacitance between the two nearly parallel plates. Capacitance readout circuitry on a nearby printed circuit board (PCB) transmits data via a low-voltage differential signaling (LVDS) interface. The fabricated SVS proof-of-concept device has successfully demonstrated approximately 1pF/gram capacitance change

HIV-1 Vpr triggers mitochondrial destruction by impairing Mfn2-mediated ER-mitochondria interaction.

PubMed

Huang, Chih-Yang; Chiang, Shu-Fen; Lin, Tze-Yi; Chiou, Shiow-Her; Chow, Kuan-Chih

2012-01-01

Human immunodeficiency virus 1 (HIV-1) viral protein R (Vpr) has been shown to induce host cell death by increasing the permeability of mitochondrial outer membrane (MOM). The mechanism underlying the damage to the mitochondria by Vpr, however, is not clearly illustrated. In this study, Vpr that is introduced, via transient transfection or lentivirus infection, into the human embryonic kidney cell line HEK293, human CD4(+) T lymphoblast cell line SupT1, or human primary CD4(+) T cells serves as the model system to study the molecular mechanism of Vpr-mediated HIV-1 pathogenesis. The results show that Vpr injures MOM and causes a loss in membrane potential (MMP) by posttranscriptionally reducing the expression of mitofusin 2 (Mfn2) via VprBP-DDB1-CUL4A ubiquitin ligase complex, gradually weakening MOM, and increasing mitochondrial deformation. Vpr also markedly decreases cytoplasmic levels of dynamin-related protein 1 (DRP1) and increases bulging in mitochondria-associated membranes (MAM), the specific regions of endoplasmic reticulum (ER) which form physical contacts with the mitochondria. Overexpression of Mfn2 and DRP1 significantly decreased the loss of MMP and apoptotic cell death caused by Vpr. Furthermore, by employing time-lapse confocal fluorescence microscopy, we identify the transport of Vpr protein from the ER, via MAM to the mitochondria. Taken together, our results suggest that Vpr-mediated cellular damage may occur on an alternative protein transport pathway from the ER, via MAM to the mitochondria, which are modulated by Mfn2 and DRP1.

HIV-1 Vpr Triggers Mitochondrial Destruction by Impairing Mfn2-Mediated ER-Mitochondria Interaction

PubMed Central

Huang, Chih-Yang; Chiang, Shu-Fen; Lin, Tze-Yi; Chiou, Shiow-Her; Chow, Kuan-Chih

2012-01-01

Human immunodeficiency virus 1 (HIV-1) viral protein R (Vpr) has been shown to induce host cell death by increasing the permeability of mitochondrial outer membrane (MOM). The mechanism underlying the damage to the mitochondria by Vpr, however, is not clearly illustrated. In this study, Vpr that is introduced, via transient transfection or lentivirus infection, into the human embryonic kidney cell line HEK293, human CD4+ T lymphoblast cell line SupT1, or human primary CD4+ T cells serves as the model system to study the molecular mechanism of Vpr-mediated HIV-1 pathogenesis. The results show that Vpr injures MOM and causes a loss in membrane potential (MMP) by posttranscriptionally reducing the expression of mitofusin 2 (Mfn2) via VprBP-DDB1-CUL4A ubiquitin ligase complex, gradually weakening MOM, and increasing mitochondrial deformation. Vpr also markedly decreases cytoplasmic levels of dynamin-related protein 1 (DRP1) and increases bulging in mitochondria-associated membranes (MAM), the specific regions of endoplasmic reticulum (ER) which form physical contacts with the mitochondria. Overexpression of Mfn2 and DRP1 significantly decreased the loss of MMP and apoptotic cell death caused by Vpr. Furthermore, by employing time-lapse confocal fluorescence microscopy, we identify the transport of Vpr protein from the ER, via MAM to the mitochondria. Taken together, our results suggest that Vpr-mediated cellular damage may occur on an alternative protein transport pathway from the ER, via MAM to the mitochondria, which are modulated by Mfn2 and DRP1. PMID:22438978

Combined Systems Approaches Reveal a Multistage Mode of Action of a Marine Antimicrobial Peptide against Pathogenic Escherichia coli and Its Protective Effect against Bacterial Peritonitis and Endotoxemia

PubMed Central

Wang, Xiumin; Teng, Da; Mao, Ruoyu; Yang, Na; Hao, Ya

2016-01-01

ABSTRACT A marine arenicin-3 derivative, N4, displayed potent antibacterial activity against Gram-negative bacteria, but its antibacterial mode of action remains elusive. The mechanism of action of N4 against pathogenic Escherichia coli was first researched by combined cytological and transcriptomic techniques in this study. The N4 peptide permeabilized the outer membrane within 1 min, disrupted the plasma membrane after 0.5 h, and localized in the cytoplasm within 5 min. Gel retardation and circular dichroism (CD) spectrum analyses demonstrated that N4 bound specifically to DNA and disrupted the DNA conformation from the B type to the C type. N4 inhibited 21.1% of the DNA and 20.6% of the RNA synthesis within 15 min. Several hallmarks of apoptosis-like cell death were exhibited by N4-induced E. coli, such as cell cycle arrest in the replication (R) and division(D) phases, reactive oxygen species production, depolarization of the plasma membrane potential, and chromatin condensation within 0.5 h. Deformed cell morphology, disappearance of the plasma membrane, leakage of the contents, and ghost cell formation were demonstrated by transmission electron microscopy, and nearly 100% of the bacteria were killed by N4. A total of 428 to 663 differentially expressed genes are involved in the response to N4, which are associated mainly with membrane biogenesis (53.9% to 56.7%) and DNA binding (13.3% to 14.9%). N4-protected mice that were lethally challenged with lipopolysaccharide (LPS) exhibited reduced levels of interleukin-6 (IL-6), IL-1β, and tumor necrosis factor alpha (TNF-α) in serum and protected the lungs from LPS-induced injury. These data facilitate an enhanced understanding of the mechanisms of marine antimicrobial peptides (AMPs) against Gram-negative bacteria and provide guidelines in developing and applying novel multitarget AMPs in the field of unlimited marine resources as therapeutics. PMID:27795369

Dynamic regulation of mitochondrial fission through modification of the dynamin-related protein Drp1

PubMed Central

Chang, Chuang-Rung; Blackstone, Craig

2017-01-01

Mitochondria in cells comprise a tubulovesicular network shaped continuously by complementary fission and fusion events. The mammalian Drp1 protein plays a key role in fission, while Mfn1, Mfn2, and OPA1 are required for fusion. Shifts in the balance between these opposing processes can occur rapidly, indicating that modifications to these proteins may regulate mitochondrial membrane dynamics. We highlight posttranslational modifications of the mitochondrial fission protein Drp1, for which these regulatory mechanisms are best characterized. This dynamin-related GTPase undergoes a number of steps to mediate mitochondrial fission, including translocation from cytoplasm to the mitochondrial outer membrane, higher-order assembly into spirals, GTP hydrolysis associated with a conformational change and membrane deformation, and ultimately disassembly. Many of these steps may be influenced by covalent modification of Drp1. We discuss the dynamic nature of Drp1 modifications and how they contribute not only to the normal regulation of mitochondrial division, but also to neuropathologic processes. PMID:20649536

Deformational injection rate measuring method

NASA Astrophysics Data System (ADS)

Marčič, Milan

2002-09-01

After completing the diesel engine endurance testing, we detected various traces of thermal load on the walls of combustion chambers located in the engine pistons. The engines were fitted with ω combustion chambers. The thermal load of different intensity levels occurred where the spray of fuel, fuel vapor, and air interacted with the combustion chamber wall. The uneven thermal load distribution of the combustion chamber wall results from varying injection rates in each injection nozzle hole. The most widely applied controlling methods so far for injection rate measurement, such as the Zeuch and Bosch concepts, allow measurement of only the total injection rate in multihole nozzles, without providing any indication whatsoever of the injection rate differences in individual injection nozzle holes. The new deformational measuring method described in the article allows the injection rate to be measured in each hole of the multihole nozzle. The results of the measurements using this method showed that the differences occurred in injection rates of individual injection nozzle holes. These differences may be the cause of various thermal loads on the combustion chamber walls. The criterion for injection rate is the deformation of the membrane due to an increase in the fuel quantity in the measuring space and due to the pressure waves resulting from the fuel being injected into the measuring space. The membrane deformation is measured using strain gauges, glued to the membrane and forming the Wheatstone's bridge. We devoted special attention to the temperature compensation of the Wheatstone's bridge and the membrane, heated up during the measurements.

Finite Element Modeling of the Deformation of a Thin Magnetoelastic Film Compared to a Membrane Model

DOE Office of Scientific and Technical Information (OSTI.GOV)

Barham, M; White, D; Steigmann, D

2009-04-08

Recently a new class of biocompatible elastic polymers loaded with small ferrous particles (magnetoelastomer) was developed at Lawrence Livermore National Laboratory. This new material was formed as a thin film using spin casting. The deformation of this material using a magnetic field has many possible applications to microfluidics. Two methods will be used to calculate the deformation of a circular magneto-elastomeric film subjected to a magnetic field. The first method is an arbitrary Lagrangian-Eulerian (ALE) finite element method (FEM) and the second is based on nonlinear continuum electromagnetism and continuum elasticity in the membrane limit. The comparison of these twomore » methods is used to test/validate the finite element method.« less

Electron microscopic study on macrogametogenesis of Eimeria labbeana infecting the Egyptian wild doves and host-parasite relationship.

PubMed

Bashtar, A R; Abdel-Ghaffar, F A; Ahmed, A K

1991-04-01

The development of macrogametes of Eimeria labbeana was studied by electron microscopy in the epithelial cells of the villi at 96 hrs. post-infection. Appearance of young macrogamonts was characterized by the loss of the architecture of the apicomplexa (polar ring, rhoptries, micronemes, conoid, subpellicular microtubules), while the pellicle became only one unit membrane. This was associated by the formation of wall forming bodies II then I. Moreover, the mitochondria, endoplasmic reticulum and Golgi were increased in the cytoplasm. Amylopectin granules as well as lipid globules were greatly increased in mature macrogametes. Host cell reaction due to infection included enlargement and deformation of infected cells, hypertrophy of their nuclei, swollen and degeneration of mitochondria, endoplasmic reticulum and vacuolation of ground cytoplasm. These changes occur in both cells with and without parasite.

Deletion of the Vaccinia Virus I2 Protein Interrupts Virion Morphogenesis, Leading to Retention of the Scaffold Protein and Mislocalization of Membrane-Associated Entry Proteins

PubMed Central

Hyun, Seong-In; Weisberg, Andrea

2017-01-01

ABSTRACT The I2L open reading frame of vaccinia virus (VACV) encodes a conserved 72-amino-acid protein with a putative C-terminal transmembrane domain. Previous studies with a tetracycline-inducible mutant demonstrated that I2-deficient virions are defective in cell entry. The purpose of the present study was to determine the step of replication or entry that is affected by loss of the I2 protein. Fluorescence microscopy experiments showed that I2 colocalized with a major membrane protein of immature and mature virions. We generated a cell line that constitutively expressed I2 and allowed construction of the VACV I2L deletion mutant vΔI2. As anticipated, vΔI2 was unable to replicate in cells that did not express I2. Unexpectedly, morphogenesis was interrupted at a stage after immature virion formation, resulting in the accumulation of dense spherical particles instead of brick-shaped mature virions with well-defined core structures. The abnormal particles retained the D13 scaffold protein of immature virions, were severely deficient in the transmembrane proteins that comprise the entry fusion complex (EFC), and had increased amounts of unprocessed membrane and core proteins. Total lysates of cells infected with vΔI2 also had diminished EFC proteins due to instability attributed to their hydrophobicity and failure to be inserted into viral membranes. A similar instability of EFC proteins had previously been found with unrelated mutants blocked earlier in morphogenesis that also accumulated viral membranes retaining the D13 scaffold. We concluded that I2 is required for virion morphogenesis, release of the D13 scaffold, and the association of EFC proteins with viral membranes. IMPORTANCE Poxviruses comprise a large family that infect vertebrates and invertebrates, cause disease in both in humans and in wild and domesticated animals, and are being engineered as vectors for vaccines and cancer therapy. In addition, investigations of poxviruses have provided insights into many aspects of cell biology. The I2 protein is conserved in all poxviruses that infect vertebrates, suggesting an important role. The present study revealed that this protein is essential for vaccinia virus morphogenesis and that its absence results in an accumulation of deformed virus particles retaining the scaffold protein and deficient in surface proteins needed for cell entry. PMID:28490596

Multiscale Morphology of Organic Semiconductor Thin Films Controls the Adhesion and Viability of Human Neural Cells

PubMed Central

Tonazzini, I.; Bystrenova, E.; Chelli, B.; Greco, P.; Stoliar, P.; Calò, A.; Lazar, A.; Borgatti, F.; D'Angelo, P.; Martini, C.; Biscarini, F.

2010-01-01

Abstract We investigate how multiscale morphology of functional thin films affects the in vitro behavior of human neural astrocytoma 1321N1 cells. Pentacene thin film morphology is precisely controlled by means of the film thickness, Θ (here expressed in monolayers (ML)). Fluorescence and atomic force microscopy allow us to correlate the shape, adhesion, and proliferation of cells to the morphological properties of pentacene films controlled by saturated roughness, σ, correlation length, ξ, and fractal dimension, df. At early incubation times, cell adhesion exhibits a transition from higher to lower values at Θ ≈ 10 ML. This is explained using a model of conformal adhesion of the cell membrane onto the growing pentacene islands. From the model fitting of the data, we show that the cell explores the surface with a deformation of the membrane whose minimum curvature radius is 90 (± 45) nm. The transition in the adhesion at ∼10 ML arises from the saturation of ξ accompanied by the monotonic increase of σ, which leads to a progressive decrease of the pentacene local radius of curvature and hence to the surface area accessible to the cell. Cell proliferation is also enhanced for Θ < 10 ML, and the optimum morphology parameter ranges for cell deployment and growth are σ ≤ 6 nm, ξ > 500 nm, and df > 2.45. The characteristic time of cell proliferation is τ ≈ 10 ± 2 h. PMID:20550892

Phospholipase Cβ1 induces membrane tubulation and is involved in caveolae formation

PubMed Central

Inaba, Takehiko; Kishimoto, Takuma; Murate, Motohide; Tajima, Takuya; Sakai, Shota; Abe, Mitsuhiro; Makino, Asami; Tomishige, Nario; Ishitsuka, Reiko; Ikeda, Yasuo; Takeoka, Shinji; Kobayashi, Toshihide

2016-01-01

Lipid membrane curvature plays important roles in various physiological phenomena. Curvature-regulated dynamic membrane remodeling is achieved by the interaction between lipids and proteins. So far, several membrane sensing/sculpting proteins, such as Bin/amphiphysin/Rvs (BAR) proteins, are reported, but there remains the possibility of the existence of unidentified membrane-deforming proteins that have not been uncovered by sequence homology. To identify new lipid membrane deformation proteins, we applied liposome-based microscopic screening, using unbiased-darkfield microscopy. Using this method, we identified phospholipase Cβ1 (PLCβ1) as a new candidate. PLCβ1 is well characterized as an enzyme catalyzing the hydrolysis of phosphatidylinositol-4,5-bisphosphate (PIP2). In addition to lipase activity, our results indicate that PLCβ1 possessed the ability of membrane tubulation. Lipase domains and inositol phospholipids binding the pleckstrin homology (PH) domain of PLCβ1 were not involved, but the C-terminal sequence was responsible for this tubulation activity. Computational modeling revealed that the C terminus displays the structural homology to the BAR domains, which is well known as a membrane sensing/sculpting domain. Overexpression of PLCβ1 caused plasma membrane tubulation, whereas knockdown of the protein reduced the number of caveolae and induced the evagination of caveolin-rich membrane domains. Taken together, our results suggest a new function of PLCβ1: plasma membrane remodeling, and in particular, caveolae formation. PMID:27342861

Dynamics of membrane nanotubes coated with I-BAR

NASA Astrophysics Data System (ADS)

Barooji, Younes F.; Rørvig-Lund, Andreas; Semsey, Szabolcs; Reihani, S. Nader S.; Bendix, Poul M.

2016-07-01

Membrane deformation is a necessary step in a number of cellular processes such as filopodia and invadopodia formation and has been shown to involve membrane shaping proteins containing membrane binding domains from the IRSp53-MIM protein family. In reconstituted membranes the membrane shaping domains can efficiently deform negatively charged membranes into tubules without any other proteins present. Here, we show that the IM domain (also called I-BAR domain) from the protein ABBA, forms semi-flexible nanotubes protruding into Giant Unilamellar lipid Vesicles (GUVs). By simultaneous quantification of tube intensity and tubular shape we find both the diameter and stiffness of the nanotubes. I-BAR decorated tubes were quantified to have a diameter of ~50 nm and exhibit no stiffening relative to protein free tubes of the same diameter. At high protein density the tubes are immobile whereas at lower density the tubes diffuse freely on the surface of the GUV. Bleaching experiments of the fluorescently tagged I-BAR confirmed that the mobility of the tubes correlates with the mobility of the I-BAR on the GUV membrane. Finally, at low density of I-BAR the protein upconcentrates within tubes protruding into the GUVs. This implies that I-BAR exhibits strong preference for negatively curved membranes.

Real-time intravital imaging of pH variation associated with osteoclast activity.

PubMed

Maeda, Hiroki; Kowada, Toshiyuki; Kikuta, Junichi; Furuya, Masayuki; Shirazaki, Mai; Mizukami, Shin; Ishii, Masaru; Kikuchi, Kazuya

2016-08-01

Intravital imaging by two-photon excitation microscopy (TPEM) has been widely used to visualize cell functions. However, small molecular probes (SMPs), commonly used for cell imaging, cannot be simply applied to intravital imaging because of the challenge of delivering them into target tissues, as well as their undesirable physicochemical properties for TPEM imaging. Here, we designed and developed a functional SMP with an active-targeting moiety, higher photostability, and a fluorescence switch and then imaged target cell activity by injecting the SMP into living mice. The combination of the rationally designed SMP with a fluorescent protein as a reporter of cell localization enabled quantitation of osteoclast activity and time-lapse imaging of its in vivo function associated with changes in cell deformation and membrane fluctuations. Real-time imaging revealed heterogenic behaviors of osteoclasts in vivo and provided insights into the mechanism of bone resorption.

Continuum electromechanical modeling of protein-membrane interactions

NASA Astrophysics Data System (ADS)

Zhou, Y. C.; Lu, Benzhuo; Gorfe, Alemayehu A.

2010-10-01

A continuum electromechanical model is proposed to describe the membrane curvature induced by electrostatic interactions in a solvated protein-membrane system. The model couples the macroscopic strain energy of membrane and the electrostatic solvation energy of the system, and equilibrium membrane deformation is obtained by minimizing the electroelastic energy functional with respect to the dielectric interface. The model is illustrated with the systems with increasing geometry complexity and captures the sensitivity of membrane curvature to the permanent and mobile charge distributions.

Conformable seal

DOEpatents

Neef, W.S.; Lambert, D.R.

1982-08-10

Sealing apparatus and method, comprising first and second surfaces or membranes, at least one of which surfaces is deformable, placed in proximity to one another. Urging means cause these surfaces to contact one another in a manner such that the deformable surface deforms to conform to the geometry of the other surface, thereby creating a seal. The seal is capable of undergoing multiple cycles of sealing and unsealing.

Active membrane masks for improved overlay performance in proximity lithography

NASA Astrophysics Data System (ADS)

Huston, Dryver R.; Plumpton, James; Esser, Brian; Sullivan, Gerald A.

2004-07-01

Membrane masks are thin (2 micron x 35 mm x 35 mm) structures that carry the master exposure patterns in proximity (X-ray) lithography. With the continuous drive to the printing of ever-finer features in microelectronics, the reduction of mask-wafer overlay positioning errors by passive rigid body positioning and passive stress control in the mask becomes impractical due to nano and sub-micron scale elastic deformations in the membrane mask. This paper describes the design, mechanics and performance of a system for actively stretching a membrane mask in-plane to control overlay distortion. The method uses thermoelectric heating/cooling elements placed on the mask perimeter. The thermoelectric elements cause controlled thermoelastic deformations in the supporting wafer, which in turn corrects distortions in the membrane mask. Silicon carbide masks are the focus of this study, but the method is believed to be applicable to other mask materials, such as diamond. Experimental and numerical results will be presented, as well as a discussion of the design issues and related design decisions.

Distinct molecular and cellular contributions to stabilizing selectin-mediated rolling under flow

PubMed Central

Yago, Tadayuki; Leppänen, Anne; Qiu, Haiying; Marcus, Warren D.; Nollert, Matthias U.; Zhu, Cheng; Cummings, Richard D.; McEver, Rodger P.

2002-01-01

Leukocytes roll on selectins at nearly constant velocities over a wide range of wall shear stresses. Ligand-coupled microspheres roll faster on selectins and detach quickly as wall shear stress is increased. To examine whether the superior performance of leukocytes reflects molecular features of native ligands or cellular properties that favor selectin-mediated rolling, we coupled structurally defined selectin ligands to microspheres or K562 cells and compared their rolling on P-selectin. Microspheres bearing soluble P-selectin glycoprotein ligand (sPSGL)-1 or 2-glycosulfopeptide (GSP)-6, a GSP modeled after the NH2-terminal P-selectin–binding region of PSGL-1, rolled equivalently but unstably on P-selectin. K562 cells displaying randomly coupled 2-GSP-6 also rolled unstably. In contrast, K562 cells bearing randomly coupled sPSGL-1 or 2-GSP-6 targeted to a membrane-distal region of the presumed glycocalyx rolled more like leukocytes: rolling steps were more uniform and shear resistant, and rolling velocities tended to plateau as wall shear stress was increased. K562 cells treated with paraformaldehyde or methyl-β-cyclodextrin before ligand coupling were less deformable and rolled unstably like microspheres. Cells treated with cytochalasin D were more deformable, further resisted detachment, and rolled slowly despite increases in wall shear stress. Thus, stable, shear-resistant rolling requires cellular properties that optimize selectin–ligand interactions. PMID:12177042

Nonlinear deformations of microcapsules in elongation flow

NASA Astrophysics Data System (ADS)

Deschamps, Julien; de Loubens, Clément; Boedec, Gwenn; Georgelin, Marc; Leonetti, Marc; Soft Matter; Biophysics Group Team

2014-11-01

Soft microcapsules are drops bounded by a thin elastic shell made of cross-linked proteins. They have numerous applications for drug delivery in bioengineering, pharmaceutics and medicine, where their mechanical stability and their dynamics under flow are crucial. They can also be used as red blood cells models. Here, we investigate the mechanical behaviour of microcapsules made of albumine in strong elongational flow, up to a stretching of 180% just before breaking. The set-up allows us to visualize the deformed shape in the two perpendicular main fields of view, to manage high capillary number and to manipulate soft microcapsules. The steady-state shape of a capsule in the planar elongational flow is non-axisymmetric. In each cross section, the shape is an ellipse but with different small axis which vary in opposite sense with the stretching. Whatever the degree of cross-linking and the size of the capsules, the deformations followed the same master-curve. Comparisons between numerical predictions and experimental results permit to conclude unambiguously that the more properly strain-energy model of membrane is the generalized Hooke model.

Interactions of the EGFR juxtamembrane domain with PIP2-containing lipid bilayers: Insights from multiscale molecular dynamics simulations☆

PubMed Central

Abd Halim, Khairul Bariyyah; Koldsø, Heidi; Sansom, Mark S.P.

2015-01-01

Background The epidermal growth factor receptor (EGFR) is the best characterised member of the receptor tyrosine kinases, which play an important role in signalling across mammalian cell membranes. The EGFR juxtamembrane (JM) domain is involved in the mechanism of activation of the receptor, interacting with the anionic lipid phosphatidylinositol 4,5-bisphosphate (PIP2) in the intracellular leaflet of the cell membrane. Methods Multiscale MD simulations were used to characterize PIP2–JM interactions. Simulations of the transmembrane helix plus JM region (TM–JM) dimer (PDB:2M20) in both PIP2-containing and PIP2-depleted lipid bilayer membranes revealed the interactions of the JM with PIP2 and other lipids. Results PIP2 forms strong interactions with the basic residues in the R645–R647 motif of the JM domain resulting in clustering of PIP2 around the protein. This association of PIP2 and the JM domain aids stabilization of JM-A dimer away from the membrane. Mutation (R645N/R646N/R647N) or PIP2-depletion results in deformation of the JM-A dimer and changes in JM–membrane interactions. Conclusions These simulations support the proposal that the positively charged residues at the start of the JM-A domain stabilize the JM-A helices in an orientation away from the membrane surface through binding to PIP2, thus promoting a conformation corresponding to an asymmetric (i.e. activated) kinase. General significance This study indicates that MD simulations may be used to characterise JM/lipid interactions, thus helping to define their role in the mechanisms of receptor tyrosine kinases. This article is part of a Special Issue entitled Recent developments of molecular dynamics. PMID:25219456

Interactions of the EGFR juxtamembrane domain with PIP2-containing lipid bilayers: Insights from multiscale molecular dynamics simulations.

PubMed

Abd Halim, Khairul Bariyyah; Koldsø, Heidi; Sansom, Mark S P

2015-05-01

The epidermal growth factor receptor (EGFR) is the best characterised member of the receptor tyrosine kinases, which play an important role in signalling across mammalian cell membranes. The EGFR juxtamembrane (JM) domain is involved in the mechanism of activation of the receptor, interacting with the anionic lipid phosphatidylinositol 4,5-bisphosphate (PIP2) in the intracellular leaflet of the cell membrane. Multiscale MD simulations were used to characterize PIP2-JM interactions. Simulations of the transmembrane helix plus JM region (TM-JM) dimer (PDB:2M20) in both PIP2-containing and PIP2-depleted lipid bilayer membranes revealed the interactions of the JM with PIP2 and other lipids. PIP2 forms strong interactions with the basic residues in the R645-R647 motif of the JM domain resulting in clustering of PIP2 around the protein. This association of PIP2 and the JM domain aids stabilization of JM-A dimer away from the membrane. Mutation (R645N/R646N/R647N) or PIP2-depletion results in deformation of the JM-A dimer and changes in JM-membrane interactions. These simulations support the proposal that the positively charged residues at the start of the JM-A domain stabilize the JM-A helices in an orientation away from the membrane surface through binding to PIP2, thus promoting a conformation corresponding to an asymmetric (i.e. activated) kinase. This study indicates that MD simulations may be used to characterise JM/lipid interactions, thus helping to define their role in the mechanisms of receptor tyrosine kinases. This article is part of a Special Issue entitled Recent developments of molecular dynamics. Copyright © 2014. Published by Elsevier B.V.

A fluid–structure interaction model to characterize bone cell stimulation in parallel-plate flow chamber systems

PubMed Central

Vaughan, T. J.; Haugh, M. G.; McNamara, L. M.

2013-01-01

Bone continuously adapts its internal structure to accommodate the functional demands of its mechanical environment and strain-induced flow of interstitial fluid is believed to be the primary mediator of mechanical stimuli to bone cells in vivo. In vitro investigations have shown that bone cells produce important biochemical signals in response to fluid flow applied using parallel-plate flow chamber (PPFC) systems. However, the exact mechanical stimulus experienced by the cells within these systems remains unclear. To fully understand this behaviour represents a most challenging multi-physics problem involving the interaction between deformable cellular structures and adjacent fluid flows. In this study, we use a fluid–structure interaction computational approach to investigate the nature of the mechanical stimulus being applied to a single osteoblast cell under fluid flow within a PPFC system. The analysis decouples the contribution of pressure and shear stress on cellular deformation and for the first time highlights that cell strain under flow is dominated by the pressure in the PPFC system rather than the applied shear stress. Furthermore, it was found that strains imparted on the cell membrane were relatively low whereas significant strain amplification occurred at the cell–substrate interface. These results suggest that strain transfer through focal attachments at the base of the cell are the primary mediators of mechanical signals to the cell under flow in a PPFC system. Such information is vital in order to correctly interpret biological responses of bone cells under in vitro stimulation and elucidate the mechanisms associated with mechanotransduction in vivo. PMID:23365189

Visualization and Sequencing of Membrane Remodeling Leading to Influenza Virus Fusion

PubMed Central

Gui, Long; Ebner, Jamie L.; Mileant, Alexander; Williams, James A.

2016-01-01

ABSTRACT Protein-mediated membrane fusion is an essential step in many fundamental biological events, including enveloped virus infection. The nature of protein and membrane intermediates and the sequence of membrane remodeling during these essential processes remain poorly understood. Here we used cryo-electron tomography (cryo-ET) to image the interplay between influenza virus and vesicles with a range of lipid compositions. By following the population kinetics of membrane fusion intermediates imaged by cryo-ET, we found that membrane remodeling commenced with the hemagglutinin fusion protein spikes grappling onto the target membrane, followed by localized target membrane dimpling as local clusters of hemagglutinin started to undergo conformational refolding. The local dimples then transitioned to extended, tightly apposed contact zones where the two proximal membrane leaflets were in most cases indistinguishable from each other, suggesting significant dehydration and possible intermingling of the lipid head groups. Increasing the content of fusion-enhancing cholesterol or bis-monoacylglycerophosphate in the target membrane led to an increase in extended contact zone formation. Interestingly, hemifused intermediates were found to be extremely rare in the influenza virus fusion system studied here, most likely reflecting the instability of this state and its rapid conversion to postfusion complexes, which increased in population over time. By tracking the populations of fusion complexes over time, the architecture and sequence of membrane reorganization leading to efficient enveloped virus fusion were thus resolved. IMPORTANCE Enveloped viruses employ specialized surface proteins to mediate fusion of cellular and viral membranes that results in the formation of pores through which the viral genetic material is delivered to the cell. For influenza virus, the trimeric hemagglutinin (HA) glycoprotein spike mediates host cell attachment and membrane fusion. While structures of a subset of conformations and parts of the fusion machinery have been characterized, the nature and sequence of membrane deformations during fusion have largely eluded characterization. Building upon studies that focused on early stages of HA-mediated membrane remodeling, here cryo-electron tomography (cryo-ET) was used to image the three-dimensional organization of intact influenza virions at different stages of fusion with liposomes, leading all the way to completion of the fusion reaction. By monitoring the evolution of fusion intermediate populations over the course of acid-induced fusion, we identified the progression of membrane reorganization that leads to efficient fusion by an enveloped virus. PMID:27226364

Membrane Mirrors With Bimorph Shape Actuators

NASA Technical Reports Server (NTRS)

Yang, Eui-Hyeok

2003-01-01

Deformable mirrors of a proposed type would be equipped with relatively-large-stroke microscopic piezoelectric actuators that would be used to maintain their reflective surfaces in precise shapes. These mirrors would be members of the class of MEMS-DM (for microelectromechanical system deformable mirror) devices, which offer potential for a precise optical control in adaptive-optics applications in such diverse fields as astronomy and vision science. The proposed mirror would be fabricated, in part, by use of a membrane-transfer technique. The actuator design would contain bimorph-type piezoelectric actuators.

Trapping, Deformation, and Rotation of Giant Unilamellar Vesicles in Octode Dielectrophoretic Field Cages

PubMed Central

Korlach, J.; Reichle, C.; Müller, T.; Schnelle, T.; Webb, W. W.

2005-01-01

The behavior of freestanding lipid bilayer membranes under the influence of dielectric force potentials was studied by trapping, holding, and rotating individual giant unilamellar vesicles (GUVs) inside dielectrophoretic microfield cages. Using laser scanning confocal microscopy and three-dimensional image reconstructions of GUVs labeled with fluorescent membrane probes, field strength and frequency-dependent vesicle deformations were observed which are explained by calculations of the dielectric force potentials inside the cage. Dynamical membrane properties under the influence of the field cage were studied by fluorescence correlation spectroscopy, circumventing potential artifacts associated with measurements involving GUV immobilization on support surfaces. Lipid transport could be accelerated markedly by the applied fields, aided by hydrodynamic fluid streaming which was also studied by fluorescence correlation spectroscopy. PMID:15863477

Structure-based membrane dome mechanism for Piezo mechanosensitivity

PubMed Central

Guo, Yusong R

2017-01-01

Mechanosensitive ion channels convert external mechanical stimuli into electrochemical signals for critical processes including touch sensation, balance, and cardiovascular regulation. The best understood mechanosensitive channel, MscL, opens a wide pore, which accounts for mechanosensitive gating due to in-plane area expansion. Eukaryotic Piezo channels have a narrow pore and therefore must capture mechanical forces to control gating in another way. We present a cryo-EM structure of mouse Piezo1 in a closed conformation at 3.7Å-resolution. The channel is a triskelion with arms consisting of repeated arrays of 4-TM structural units surrounding a pore. Its shape deforms the membrane locally into a dome. We present a hypothesis in which the membrane deformation changes upon channel opening. Quantitatively, membrane tension will alter gating energetics in proportion to the change in projected area under the dome. This mechanism can account for highly sensitive mechanical gating in the setting of a narrow, cation-selective pore. PMID:29231809

The origin of the vertebrate skeleton

NASA Astrophysics Data System (ADS)

Pivar, Stuart

2011-01-01

The anatomy of the human and other vertebrates has been well described since the days of Leonardo da Vinci and Vesalius. The causative origin of the configuration of the bones and of their shapes and forms has been addressed over the ensuing centuries by such outstanding investigators as Goethe, Von Baer, Gegenbauer, Wilhelm His and D'Arcy Thompson, who sought to apply mechanical principles to morphogenesis. However, no coherent causative model of morphogenesis has ever been presented. This paper presents a causative model for the origin of the vertebrate skeleton, based on the premise that the body is a mosaic enlargement of self-organized patterns engrained in the membrane of the egg cell. Drawings illustrate the proposed hypothetical origin of membrane patterning and the changes in the hydrostatic equilibrium of the cytoplasm that cause topographical deformations resulting in the vertebrate body form.

Phase geometries of two-dimensional excitable waves govern self-organized morphodynamics of amoeboid cells

PubMed Central

Taniguchi, Daisuke; Ishihara, Shuji; Oonuki, Takehiko; Honda-Kitahara, Mai; Kaneko, Kunihiko; Sawai, Satoshi

2013-01-01

In both randomly moving Dictyostelium and mammalian cells, phosphatidylinositol (3,4,5)-trisphosphate and F-actin are known to propagate as waves at the membrane and act to push out the protruding edge. To date, however, the relationship between the wave geometry and the patterns of amoeboid shape change remains elusive. Here, by using phase map analysis, we show that morphology dynamics of randomly moving Dictyostelium discoideum cells can be characterized by the number, topology, and position of spatial phase singularities, i.e., points that represent organizing centers of rotating waves. A single isolated singularity near the cellular edge induced a rotational protrusion, whereas a pair of singularities supported a symmetric extension. These singularities appeared by strong phase resetting due to de novo nucleation at the back of preexisting waves. Analysis of a theoretical model indicated excitability of the system that is governed by positive feedback from phosphatidylinositol (3,4,5)-trisphosphate to PI3-kinase activation, and we showed experimentally that this requires F-actin. Furthermore, by incorporating membrane deformation into the model, we demonstrated that geometries of competing waves explain most of the observed semiperiodic changes in amoeboid morphology. PMID:23479620

Hydrogen separation process

DOEpatents

Mundschau, Michael [Longmont, CO; Xie, Xiaobing [Foster City, CA; Evenson, IV, Carl; Grimmer, Paul [Longmont, CO; Wright, Harold [Longmont, CO

2011-05-24

A method for separating a hydrogen-rich product stream from a feed stream comprising hydrogen and at least one carbon-containing gas, comprising feeding the feed stream, at an inlet pressure greater than atmospheric pressure and a temperature greater than 200.degree. C., to a hydrogen separation membrane system comprising a membrane that is selectively permeable to hydrogen, and producing a hydrogen-rich permeate product stream on the permeate side of the membrane and a carbon dioxide-rich product raffinate stream on the raffinate side of the membrane. A method for separating a hydrogen-rich product stream from a feed stream comprising hydrogen and at least one carbon-containing gas, comprising feeding the feed stream, at an inlet pressure greater than atmospheric pressure and a temperature greater than 200.degree. C., to an integrated water gas shift/hydrogen separation membrane system wherein the hydrogen separation membrane system comprises a membrane that is selectively permeable to hydrogen, and producing a hydrogen-rich permeate product stream on the permeate side of the membrane and a carbon dioxide-rich product raffinate stream on the raffinate side of the membrane. A method for pretreating a membrane, comprising: heating the membrane to a desired operating temperature and desired feed pressure in a flow of inert gas for a sufficient time to cause the membrane to mechanically deform; decreasing the feed pressure to approximately ambient pressure; and optionally, flowing an oxidizing agent across the membrane before, during, or after deformation of the membrane. A method of supporting a hydrogen separation membrane system comprising selecting a hydrogen separation membrane system comprising one or more catalyst outer layers deposited on a hydrogen transport membrane layer and sealing the hydrogen separation membrane system to a porous support.

The ESCRT-III pathway facilitates cardiomyocyte release of cBIN1-containing microparticles.

PubMed

Xu, Bing; Fu, Ying; Liu, Yan; Agvanian, Sosse; Wirka, Robert C; Baum, Rachel; Zhou, Kang; Shaw, Robin M; Hong, TingTing

2017-08-01

Microparticles (MPs) are cell-cell communication vesicles derived from the cell surface plasma membrane, although they are not known to originate from cardiac ventricular muscle. In ventricular cardiomyocytes, the membrane deformation protein cardiac bridging integrator 1 (cBIN1 or BIN1+13+17) creates transverse-tubule (t-tubule) membrane microfolds, which facilitate ion channel trafficking and modulate local ionic concentrations. The microfold-generated microdomains continuously reorganize, adapting in response to stress to modulate the calcium signaling apparatus. We explored the possibility that cBIN1-microfolds are externally released from cardiomyocytes. Using electron microscopy imaging with immunogold labeling, we found in mouse plasma that cBIN1 exists in membrane vesicles about 200 nm in size, which is consistent with the size of MPs. In mice with cardiac-specific heterozygous Bin1 deletion, flow cytometry identified 47% less cBIN1-MPs in plasma, supporting cardiac origin. Cardiac release was also evidenced by the detection of cBIN1-MPs in medium bathing a pure population of isolated adult mouse cardiomyocytes. In human plasma, osmotic shock increased cBIN1 detection by enzyme-linked immunosorbent assay (ELISA), and cBIN1 level decreased in humans with heart failure, a condition with reduced cardiac muscle cBIN1, both of which support cBIN1 release in MPs from human hearts. Exploring putative mechanisms of MP release, we found that the membrane fission complex endosomal sorting complexes required for transport (ESCRT)-III subunit charged multivesicular body protein 4B (CHMP4B) colocalizes and coimmunoprecipitates with cBIN1, an interaction enhanced by actin stabilization. In HeLa cells with cBIN1 overexpression, knockdown of CHMP4B reduced the release of cBIN1-MPs. Using truncation mutants, we identified that the N-terminal BAR (N-BAR) domain in cBIN1 is required for CHMP4B binding and MP release. This study links the BAR protein superfamily to the ESCRT pathway for MP biogenesis in mammalian cardiac ventricular cells, identifying elements of a pathway by which cytoplasmic cBIN1 is released into blood.

Sealing apparatus utilizing a conformable member

DOEpatents

Neef, William S.; Lambert, Donald R.

1988-01-01

Sealing apparatus and method, comprising first and second surfaces or membranes, at least one of which surfaces is deformable, placed in proximity to one another. Urging means cause these surfaces to contact one another in a manner such that the deformable surface "deforms" to conform to the geometry of the other surface, thereby creating a seal. The seal is capable of undergoing multiple cycles of sealing and unsealing.

Distribution of protein motors along the lateral wall of the outer hair cell.

PubMed

Wada, H; Usukura, H; Takeuchi, S; Sugawara, M; Kakehata, S; Ikeda, K

2001-12-01

The outer hair cell (OHC) plays an important role in the normal functioning of the cochlea, and cochlear amplification is believed to be based on OHC electromotility. This electromotility putatively arises from a conformational change of molecules, i.e., 'protein motors', which would be distributed along the plasma membrane. Although it has been assumed that protein motors are distributed in a restricted area of the plasma membrane, details of such distribution remain unclarified. In this study, first, in order to understand the difference in the stiffness along the cell axis, the local deformation of the OHC in response to hypotonic stimulation is analyzed by measuring the displacement of microspheres attached randomly to the lateral wall of the cell. As a result, the stiffness is expected to be constant throughout the region except in the apical part of the cell, and the stiffness of the apical part is expected to be higher than that of the other regions. Then, the local elongation and contraction of the OHC in response to sinusoidal voltage stimulation are analyzed by measuring the displacement of the microspheres in the same way as in the case of the hypotonic stimulation. From the two measurements mentioned above, it is concluded that there are no motors in the apical and basal parts of the cell, and that the motors are equally distributed along the cell lateral wall in the middle part of the cell.

Effects of an angelica extract on human erythrocyte aggregation, deformation and osmotic fragility.

PubMed

Wang, X; Wei, L; Ouyang, J P; Muller, S; Gentils, M; Cauchois, G; Stoltz, J F

2001-01-01

In Chinese traditional medicine, angelica is widely used for its known clinical effects of ameliorating blood microcirculation. But the mechanism of these beneficial effects still remains unclear. In this work the rheological behaviour of human erythrocytes treated by angelica was studied in vitro. Normal RBCs incubated with an angelica extract at different concentrations (5, 10 or 20 mg/ml) for 60 min at 37 degrees C and then their aggregation, deformation and osmotic fragility were measured with different recently developed optical techniques, namely Erythroaggregometer (Regulest, Florange, France), LORCA (Mechatronics, Amsterdam) and Fragilimeter (Regulest, Florange, France). Experimental results show that angelica (20 mg/ml) significantly decreased normal RBCs' aggregation speed (p<0.01) and could inhibit the hyperaggregability caused by dextran 500. However, the strength of normal RBCs aggregates were not influenced by angelica. When a calcium ionophore A23187 (1.9 microM) was used to harden cell membrane, angelica (20 mg/ml) could significantly (p<0.01) protect erythrocytes against the loss of their deformability even it had no effects on normal RBCs deformation. Finally angelica (5 and 10 mg/ml) decreased significantly (p<0.01) normal RBCs osmotic fragility. In conclusion angelica plays a rheologically active role on human erythrocytes, and this study suggests a possible mechanism for angelica's positive effects against certain cardiovascular diseases.

Lightweight In-Plane Actuated Deformable Mirrors for Space Telescopes

DTIC Science & Technology

2006-09-01

dimensional beam-string and axisymmetric plate-membrane. The beam-string (a clamped beam simultaneously under an axial load ) is an important...Tensile load versus radius. . . . . . . . . . . . . . . . . . . . . . 175 7.4. Actuation voltage functions. . . . . . . . . . . . . . . . . . . . 179...membrane Asymptotic finite element Flint and De- noyer [45] 2003 In-plane Circular membrane Numerical least squares fit Actuators modelled as line loads

Fabrication Process of Silicone-based Dielectric Elastomer Actuators

PubMed Central

Rosset, Samuel; Araromi, Oluwaseun A.; Schlatter, Samuel; Shea, Herbert R.

2016-01-01

This contribution demonstrates the fabrication process of dielectric elastomer transducers (DETs). DETs are stretchable capacitors consisting of an elastomeric dielectric membrane sandwiched between two compliant electrodes. The large actuation strains of these transducers when used as actuators (over 300% area strain) and their soft and compliant nature has been exploited for a wide range of applications, including electrically tunable optics, haptic feedback devices, wave-energy harvesting, deformable cell-culture devices, compliant grippers, and propulsion of a bio-inspired fish-like airship. In most cases, DETs are made with a commercial proprietary acrylic elastomer and with hand-applied electrodes of carbon powder or carbon grease. This combination leads to non-reproducible and slow actuators exhibiting viscoelastic creep and a short lifetime. We present here a complete process flow for the reproducible fabrication of DETs based on thin elastomeric silicone films, including casting of thin silicone membranes, membrane release and prestretching, patterning of robust compliant electrodes, assembly and testing. The membranes are cast on flexible polyethylene terephthalate (PET) substrates coated with a water-soluble sacrificial layer for ease of release. The electrodes consist of carbon black particles dispersed into a silicone matrix and patterned using a stamping technique, which leads to precisely-defined compliant electrodes that present a high adhesion to the dielectric membrane on which they are applied. PMID:26863283

Bitter taste receptor agonists alter mitochondrial function and induce autophagy in airway smooth muscle cells.

PubMed

Pan, Shi; Sharma, Pawan; Shah, Sushrut D; Deshpande, Deepak A

2017-07-01

Airway remodeling, including increased airway smooth muscle (ASM) mass, is a hallmark feature of asthma and COPD. We previously identified the expression of bitter taste receptors (TAS2Rs) on human ASM cells and demonstrated that known TAS2R agonists could promote ASM relaxation and bronchodilation and inhibit mitogen-induced ASM growth. In this study, we explored cellular mechanisms mediating the antimitogenic effect of TAS2R agonists on human ASM cells. Pretreatment of ASM cells with TAS2R agonists chloroquine and quinine resulted in inhibition of cell survival, which was largely reversed by bafilomycin A1, an autophagy inhibitor. Transmission electron microscope studies demonstrated the presence of double-membrane autophagosomes and deformed mitochondria. In ASM cells, TAS2R agonists decreased mitochondrial membrane potential and increased mitochondrial ROS and mitochondrial fragmentation. Inhibiting dynamin-like protein 1 (DLP1) reversed TAS2R agonist-induced mitochondrial membrane potential change and attenuated mitochondrial fragmentation and cell death. Furthermore, the expression of mitochondrial protein BCL2/adenovirus E1B 19-kDa protein-interacting protein 3 (Bnip3) and mitochondrial localization of DLP1 were significantly upregulated by TAS2R agonists. More importantly, inhibiting Bnip3 mitochondrial localization by dominant-negative Bnip3 significantly attenuated cell death induced by TAS2R agonist. Collectively the TAS2R agonists chloroquine and quinine modulate mitochondrial structure and function, resulting in ASM cell death. Furthermore, Bnip3 plays a central role in TAS2R agonist-induced ASM functional changes via a mitochondrial pathway. These findings further establish the cellular mechanisms of antimitogenic effects of TAS2R agonists and identify a novel class of receptors and pathways that can be targeted to mitigate airway remodeling as well as bronchoconstriction in obstructive airway diseases. Copyright © 2017 the American Physiological Society.

A tethering complex drives the terminal stage of SNARE-dependent membrane fusion

NASA Astrophysics Data System (ADS)

D'Agostino, Massimo; Risselada, Herre Jelger; Lürick, Anna; Ungermann, Christian; Mayer, Andreas

2017-11-01

Membrane fusion in eukaryotic cells mediates the biogenesis of organelles, vesicular traffic between them, and exo- and endocytosis of important signalling molecules, such as hormones and neurotransmitters. Distinct tasks in intracellular membrane fusion have been assigned to conserved protein systems. Tethering proteins mediate the initial recognition and attachment of membranes, whereas SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) protein complexes are considered as the core fusion engine. SNARE complexes provide mechanical energy to distort membranes and drive them through a hemifusion intermediate towards the formation of a fusion pore. This last step is highly energy-demanding. Here we combine the in vivo and in vitro fusion of yeast vacuoles with molecular simulations to show that tethering proteins are critical for overcoming the final energy barrier to fusion pore formation. SNAREs alone drive vacuoles only into the hemifused state. Tethering proteins greatly increase the volume of SNARE complexes and deform the site of hemifusion, which lowers the energy barrier for pore opening and provides the driving force. Thereby, tethering proteins assume a crucial mechanical role in the terminal stage of membrane fusion that is likely to be conserved at multiple steps of vesicular traffic. We therefore propose that SNAREs and tethering proteins should be considered as a single, non-dissociable device that drives fusion. The core fusion machinery may then be larger and more complex than previously thought.

Equilibrium electrodeformation of a spheroidal vesicle in an ac electric field

NASA Astrophysics Data System (ADS)

Nganguia, H.; Young, Y.-N.

2013-11-01

In this work, we develop a theoretical model to explain the equilibrium spheroidal deformation of a giant unilamellar vesicle (GUV) under an alternating (ac) electric field. Suspended in a leaky dielectric fluid, the vesicle membrane is modeled as a thin capacitive spheroidal shell. The equilibrium vesicle shape results from the balance between mechanical forces from the viscous fluid, the restoring elastic membrane forces, and the externally imposed electric forces. Our spheroidal model predicts a deformation-dependent transmembrane potential, and is able to capture large deformation of a vesicle under an electric field. A detailed comparison against both experiments and small-deformation (quasispherical) theory showed that the spheroidal model gives better agreement with experiments in terms of the dependence on fluid conductivity ratio, permittivity ratio, vesicle size, electric field strength, and frequency. The spheroidal model also allows for an asymptotic analysis on the crossover frequency where the equilibrium vesicle shape crosses over between prolate and oblate shapes. Comparisons show that the spheroidal model gives better agreement with experimental observations.

Liquid gating elastomeric porous system with dynamically controllable gas/liquid transport.

PubMed

Sheng, Zhizhi; Wang, Honglong; Tang, Yongliang; Wang, Miao; Huang, Lizhi; Min, Lingli; Meng, Haiqiang; Chen, Songyue; Jiang, Lei; Hou, Xu

2018-02-01

The development of membrane technology is central to fields ranging from resource harvesting to medicine, but the existing designs are unable to handle the complex sorting of multiphase substances required for many systems. Especially, the dynamic multiphase transport and separation under a steady-state applied pressure have great benefits for membrane science, but have not been realized at present. Moreover, the incorporation of precisely dynamic control with avoidance of contamination of membranes remains elusive. We show a versatile strategy for creating elastomeric microporous membrane-based systems that can finely control and dynamically modulate the sorting of a wide range of gases and liquids under a steady-state applied pressure, nearly eliminate fouling, and can be easily applied over many size scales, pressures, and environments. Experiments and theoretical calculation demonstrate the stability of our system and the tunability of the critical pressure. Dynamic transport of gas and liquid can be achieved through our gating interfacial design and the controllable pores' deformation without changing the applied pressure. Therefore, we believe that this system will bring new opportunities for many applications, such as gas-involved chemical reactions, fuel cells, multiphase separation, multiphase flow, multiphase microreactors, colloidal particle synthesis, and sizing nano/microparticles.

Liquid gating elastomeric porous system with dynamically controllable gas/liquid transport

PubMed Central

Sheng, Zhizhi; Wang, Honglong; Tang, Yongliang; Wang, Miao; Huang, Lizhi; Min, Lingli; Meng, Haiqiang; Chen, Songyue; Jiang, Lei; Hou, Xu

2018-01-01

The development of membrane technology is central to fields ranging from resource harvesting to medicine, but the existing designs are unable to handle the complex sorting of multiphase substances required for many systems. Especially, the dynamic multiphase transport and separation under a steady-state applied pressure have great benefits for membrane science, but have not been realized at present. Moreover, the incorporation of precisely dynamic control with avoidance of contamination of membranes remains elusive. We show a versatile strategy for creating elastomeric microporous membrane-based systems that can finely control and dynamically modulate the sorting of a wide range of gases and liquids under a steady-state applied pressure, nearly eliminate fouling, and can be easily applied over many size scales, pressures, and environments. Experiments and theoretical calculation demonstrate the stability of our system and the tunability of the critical pressure. Dynamic transport of gas and liquid can be achieved through our gating interfacial design and the controllable pores’ deformation without changing the applied pressure. Therefore, we believe that this system will bring new opportunities for many applications, such as gas-involved chemical reactions, fuel cells, multiphase separation, multiphase flow, multiphase microreactors, colloidal particle synthesis, and sizing nano/microparticles. PMID:29487906

Extracting Cell Stiffness from Real-Time Deformability Cytometry: Theory and Experiment.

PubMed

Mietke, Alexander; Otto, Oliver; Girardo, Salvatore; Rosendahl, Philipp; Taubenberger, Anna; Golfier, Stefan; Ulbricht, Elke; Aland, Sebastian; Guck, Jochen; Fischer-Friedrich, Elisabeth

2015-11-17

Cell stiffness is a sensitive indicator of physiological and pathological changes in cells, with many potential applications in biology and medicine. A new method, real-time deformability cytometry, probes cell stiffness at high throughput by exposing cells to a shear flow in a microfluidic channel, allowing for mechanical phenotyping based on single-cell deformability. However, observed deformations of cells in the channel not only are determined by cell stiffness, but also depend on cell size relative to channel size. Here, we disentangle mutual contributions of cell size and cell stiffness to cell deformation by a theoretical analysis in terms of hydrodynamics and linear elasticity theory. Performing real-time deformability cytometry experiments on both model spheres of known elasticity and biological cells, we demonstrate that our analytical model not only predicts deformed shapes inside the channel but also allows for quantification of cell mechanical parameters. Thereby, fast and quantitative mechanical sampling of large cell populations becomes feasible. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.

Extracting Cell Stiffness from Real-Time Deformability Cytometry: Theory and Experiment

PubMed Central

Mietke, Alexander; Otto, Oliver; Girardo, Salvatore; Rosendahl, Philipp; Taubenberger, Anna; Golfier, Stefan; Ulbricht, Elke; Aland, Sebastian; Guck, Jochen; Fischer-Friedrich, Elisabeth

2015-01-01

Cell stiffness is a sensitive indicator of physiological and pathological changes in cells, with many potential applications in biology and medicine. A new method, real-time deformability cytometry, probes cell stiffness at high throughput by exposing cells to a shear flow in a microfluidic channel, allowing for mechanical phenotyping based on single-cell deformability. However, observed deformations of cells in the channel not only are determined by cell stiffness, but also depend on cell size relative to channel size. Here, we disentangle mutual contributions of cell size and cell stiffness to cell deformation by a theoretical analysis in terms of hydrodynamics and linear elasticity theory. Performing real-time deformability cytometry experiments on both model spheres of known elasticity and biological cells, we demonstrate that our analytical model not only predicts deformed shapes inside the channel but also allows for quantification of cell mechanical parameters. Thereby, fast and quantitative mechanical sampling of large cell populations becomes feasible. PMID:26588562

Electrothermal Equivalent Three-Dimensional Finite-Element Model of a Single Neuron.

PubMed

Cinelli, Ilaria; Destrade, Michel; Duffy, Maeve; McHugh, Peter

2018-06-01

We propose a novel approach for modelling the interdependence of electrical and mechanical phenomena in nervous cells, by using electrothermal equivalences in finite element (FE) analysis so that existing thermomechanical tools can be applied. First, the equivalence between electrical and thermal properties of the nerve materials is established, and results of a pure heat conduction analysis performed in Abaqus CAE Software 6.13-3 are validated with analytical solutions for a range of steady and transient conditions. This validation includes the definition of equivalent active membrane properties that enable prediction of the action potential. Then, as a step toward fully coupled models, electromechanical coupling is implemented through the definition of equivalent piezoelectric properties of the nerve membrane using the thermal expansion coefficient, enabling prediction of the mechanical response of the nerve to the action potential. Results of the coupled electromechanical model are validated with previously published experimental results of deformation for squid giant axon, crab nerve fibre, and garfish olfactory nerve fibre. A simplified coupled electromechanical modelling approach is established through an electrothermal equivalent FE model of a nervous cell for biomedical applications. One of the key findings is the mechanical characterization of the neural activity in a coupled electromechanical domain, which provides insights into the electromechanical behaviour of nervous cells, such as thinning of the membrane. This is a first step toward modelling three-dimensional electromechanical alteration induced by trauma at nerve bundle, tissue, and organ levels.

How sterol tilt regulates properties and organization of lipid membranes and membrane insertions

PubMed Central

Khelashvili, George; Harries, Daniel

2013-01-01

Serving as a crucial component of mammalian cells, cholesterol critically regulates the functions of biomembranes. This review focuses on a specific property of cholesterol and other sterols: the tilt modulus χ that quantifies the energetic cost of tilting sterol molecules inside the lipid membrane. We show how χ is involved in determining properties of cholesterol-containing membranes, and detail a novel approach to quantify its value from atomistic molecular dynamics (MD) simulations. Specifically, we link χ with other structural, thermodynamic, and mechanical properties of cholesterol-containing lipid membranes, and delineate how this useful parameter can be obtained from the sterol tilt probability distributions derived from relatively small-scale unbiased MD simulations. We demonstrate how the tilt modulus quantitatively describes the aligning field that sterol molecules create inside the phospholipid bilayers, and we relate χ to the bending rigidity of the lipid bilayer through effective tilt and splay energy contributions to the elastic deformations. Moreover, we show how χ can conveniently characterize the “condensing effect” of cholesterol on phospholipids. Finally, we demonstrate the importance of this cholesterol aligning field to the proper folding and interactions of membrane peptides. Given the relative ease of obtaining the tilt modulus from atomistic simulations, we propose that χ can be routinely used to characterize the mechanical properties of sterol/lipid bilayers, and can also serve as a required fitting parameter in multi-scaled simulations of lipid membrane models to relate the different levels of coarse-grained details. PMID:23291283

Phase imaging microscopy for the diagnostics of plasma-cell interaction

NASA Astrophysics Data System (ADS)

Ohene, Yolanda; Marinov, Ilya; de Laulanié, Lucie; Dupuy, Corinne; Wattelier, Benoit; Starikovskaia, Svetlana

2015-06-01

Phase images of biological specimens were obtained by the method of Quadriwave Lateral Shearing Interferometry (QWLSI). The QWLSI technique produces, at high resolution, phase images of the cells having been exposed to a plasma treatment and enables the quantitative analysis of the changes in the surface area of the cells over time. Morphological changes in the HTori normal thyroid cells were demonstrated using this method. There was a comparison of the cell behaviour between control cells, cells treated by plasma of a nanosecond dielectric barrier discharge, including cells pre-treated by catalase, and cells treated with an equivalent amount of H2O2. The major changes in the cell membrane morphology were observed at only 5 min after the plasma treatment. The primary role of reactive oxygen species (ROS) in this degradation is suggested. Deformation and condensation of the cell nucleus were observed 2-3 h after the treatment and are supposedly related to apoptosis induction. The coupling of the phase QWLSI with immunofluorescence imaging would give a deeper insight into the mechanisms of plasma induced cell death.

Cell mechanics in biomedical cavitation

PubMed Central

Wang, Qianxi; Manmi, Kawa; Liu, Kuo-Kang

2015-01-01

Studies on the deformation behaviours of cellular entities, such as coated microbubbles and liposomes subject to a cavitation flow, become increasingly important for the advancement of ultrasonic imaging and drug delivery. Numerical simulations for bubble dynamics of ultrasound contrast agents based on the boundary integral method are presented in this work. The effects of the encapsulating shell are estimated by adapting Hoff's model used for thin-shell contrast agents. The viscosity effects are estimated by including the normal viscous stress in the boundary condition. In parallel, mechanical models of cell membranes and liposomes as well as state-of-the-art techniques for quantitative measurement of viscoelasticity for a single cell or coated microbubbles are reviewed. The future developments regarding modelling and measurement of the material properties of the cellular entities for cutting-edge biomedical applications are also discussed. PMID:26442142

Role of RhoA and its effectors ROCK and mDia1 in the modulation of deformation-induced FAK, ERK, p38, and MLC motogenic signals in human Caco-2 intestinal epithelial cells

PubMed Central

Chaturvedi, Lakshmi S.; Marsh, Harold M.

2011-01-01

Repetitive deformation enhances intestinal epithelial migration across tissue fibronectin. We evaluated the contribution of RhoA and its effectors Rho-associated kinase (ROK/ROCK) and mammalian diaphanous formins (mDia1) to deformation-induced intestinal epithelial motility across fibronectin and the responsible focal adhesion kinase (FAK), extracellular signal-regulated kinase (ERK), p38, and myosin light chain (MLC) signaling. We reduced RhoA, ROCK1, ROCK2, and mDia1 by smart-pool double-stranded short-interfering RNAs (siRNA) and pharmacologically inhibited RhoA, ROCK, and FAK in human Caco-2 intestinal epithelial monolayers on fibronectin-coated membranes subjected to 10% repetitive deformation at 10 cycles/min. Migration was measured by wound closure. Stimulation of migration by deformation was prevented by exoenzyme C3, Y27632, or selective RhoA, ROCK1, and ROCK2 or mDia1 siRNAs. RhoA, ROCK inhibition, or RhoA, ROCK1, ROCK2, mDia1, and FAK reduction by siRNA blocked deformation-induced nuclear ERK phosphorylation without preventing ERK phosphorylation in the cytoplasmic protein fraction. Furthermore, RhoA, ROCK inhibition or RhoA, ROCK1, ROCK2, and mDia1 reduction by siRNA also blocked strain-induced FAK-Tyr925, p38, and MLC phosphorylation. These results suggest that RhoA, ROCK, mDia1, FAK, ERK, p38, and MLC all mediate the stimulation of intestinal epithelial migration by repetitive deformation. This pathway may be an important target for interventions to promote mechanotransduced mucosal healing during inflammation. PMID:21849669

A continuum membrane model for small deformations of a spider orb-web

NASA Astrophysics Data System (ADS)

Morassi, Antonino; Soler, Alejandro; Zaera, Ramón

2017-09-01

In this paper we propose a continuum membrane model for the infinitesimal deformation of a spider web. The model is derived in the simple context of axially-symmetric webs formed by radial threads connected with circumferential threads belonging to concentric circles. Under suitable assumption on the tensile pre-stress acting in the referential configuration, the out-of-plane static equilibrium and the free transverse and in-plane vibration of a supported circular orb-web are studied in detail. The accuracy of the model in describing a discrete spider web is numerically investigated.

Fragmentation of cancer cells

NASA Astrophysics Data System (ADS)

Vanapalli, Siva; Kamyabi, Nabiollah

Tumor cells have to travel through blood capillaries to be able to metastasize and colonize in distant organs. Among the numerous cells that are shed by the primary tumor, very few survive in circulation. In vivo studies have shown that tumor cells can undergo breakup at microcapillary junctions affecting their survival. It is currently unclear what hydrodynamic and biomechanical factors contribute to fragmentation and moreover how different are the breakup dynamics of highly and weakly metastatic cells. In this study, we use microfluidics to investigate flow-induced breakup of prostate and breast cancer cells. We observe several different modes of breakup of cancer cells, which have striking similarities with breakup of viscous drops. We quantify the breakup time and find that highly metastatic cancer cells take longer to breakup than lowly metastatic cells suggesting that tumor cells may dynamically modify their deformability to avoid fragmentation. We also identify the role that cytoskeleton and membrane plays in the breakup process. Our study highlights the important role that tumor cell fragmentation plays in cancer metastasis. Cancer Prevention and Research Institute of Texas.

Protozoa inhibition by different salts: Osmotic stress or ionic stress?

PubMed

Li, Changhao; Li, Jingya; Lan, Christopher Q; Liao, Dankui

2017-09-01

Cell density and morphology changes were tested to examine the effects of salts including NaHCO 3 , NaCl, KHCO 3 , and KCl at 160 mM on protozoa. It was demonstrated that ionic stress rather than osmotic stress led to protozoa cell death and NaHCO 3 was shown to be the most effective inhibitor. Deformation of cells and cell shrinkage were observed when protozoan cells were exposed to polyethylene glycol (PEG) or any of the salts. However, while PEG treated cells could fully recover in both number and size, only a small portion of the salt-treated cells survive and cell size was 36-58% smaller than the regular. The disappearance of salt-treated protozoa cells was hypothetically attributed to disruption of the cytoplasmic membrane of these cells. It is further hypothesized that the PEG-treated protozoan cells carried out regulatory volume increase (RVI) after the osmotic shock but the RVI of salt-treated protozoa was hurdled to varied extents. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1418-1424, 2017. © 2017 American Institute of Chemical Engineers.

A deformation model of flexible, HAMR objects for accurate propagation under perturbations and the self-shadowing effects

NASA Astrophysics Data System (ADS)

Channumsin, Sittiporn; Ceriotti, Matteo; Radice, Gianmarco

2018-02-01

A new type of space debris in near geosynchronous orbit (GEO) was recently discovered and later identified as exhibiting unique characteristics associated with high area-to-mass ratio (HAMR) objects, such as high rotation rates and high reflection properties. Observations have shown that this debris type is very sensitive to environmental disturbances, particularly solar radiation pressure, due to the fact that its motion depends on the actual effective area, orientation of that effective area, reflection properties and the area-to-mass ratio of the object is not stable over time. Previous investigations have modelled this type of debris as rigid bodies (constant area-to-mass ratios) or discrete deformed body; however, these simplifications will lead to inaccurate long term orbital predictions. This paper proposes a simple yet reliable model of a thin, deformable membrane based on multibody dynamics. The membrane is modelled as a series of flat plates, connected through joints, representing the flexibility of the membrane itself. The mass of the membrane, albeit low, is taken into account through lump masses at the joints. The attitude and orbital motion of this flexible membrane model is then propagated near GEO to predict its orbital evolution under the perturbations of solar radiation pressure, Earth's gravity field (J2), third body gravitational fields (the Sun and Moon) and self-shadowing. These results are then compared to those obtained for two rigid body models (cannonball and flat rigid plate). In addition, Monte Carlo simulations of the flexible model by varying initial attitude and deformation angle (different shape) are investigated and compared with the two rigid models (cannonball and flat rigid plate) over a period of 100 days. The numerical results demonstrate that cannonball and rigid flat plate are not appropriate to capture the true dynamical evolution of these objects, at the cost of increased computational time.

A universal piezo-driven ultrasonic cell microinjection system.

PubMed

Huang, Haibo; Mills, James K; Lu, Cong; Sun, Dong

2011-08-01

Over the past decade, the rapid development of biotechnologies such as gene injection, in-vitro fertilization, intracytoplasmic sperm injection (ICSI) and drug development have led to great demand for highly automated, high precision equipment for microinjection. Recently a new cell injection technology using piezo-driven pipettes with a very small mercury column was proposed and successfully applied in ICSI to a variety of mammal species. Although this technique significantly improves the survival rates of the ICSI process, shortcomings due to the toxicity of mercury and damage to the cell membrane due to large lateral tip oscillations of the injector pipette may limit its application. In this paper, a new cell injection system for automatic batch injection of suspended cells is developed. A new design of the piezo-driven cell injector is proposed for automated suspended cell injection. This new piezo-driven cell injector design relocates the piezo oscillation actuator to the injector pipette which eliminates the vibration effect on other parts of the micromanipulator. A small piezo stack is sufficient to perform the cell injection process. Harmful lateral tip oscillations of the injector pipette are reduced substantially without the use of a mercury column. Furthermore, ultrasonic vibration micro-dissection (UVM) theory is utilized to analyze the piezo-driven cell injection process, and the source of the lateral oscillations of the injector pipette is investigated. From preliminary experiments of cell injection of a large number of zebrafish embryos (n = 200), the injector pipette can easily pierce through the cell membrane at a low injection speed and almost no deformation of the cell wall, and with a high success rate(96%) and survival rate(80.7%) This new injection approach shows good potential for precision injection with less damage to the injected cells.

Biophysics of malarial parasite exit from infected erythrocytes.

PubMed

Chandramohanadas, Rajesh; Park, YongKeun; Lui, Lena; Li, Ang; Quinn, David; Liew, Kingsley; Diez-Silva, Monica; Sung, Yongjin; Dao, Ming; Lim, Chwee Teck; Preiser, Peter Rainer; Suresh, Subra

2011-01-01

Upon infection and development within human erythrocytes, P. falciparum induces alterations to the infected RBC morphology and bio-mechanical properties to eventually rupture the host cells through parasitic and host derived proteases of cysteine and serine families. We used previously reported broad-spectrum inhibitors (E64d, EGTA-AM and chymostatin) to inhibit these proteases and impede rupture to analyze mechanical signatures associated with parasite escape. Treatment of late-stage iRBCs with E64d and EGTA-AM prevented rupture, resulted in no major RBC cytoskeletal reconfiguration but altered schizont morphology followed by dramatic re-distribution of three-dimensional refractive index (3D-RI) within the iRBC. These phenotypes demonstrated several-fold increased iRBC membrane flickering. In contrast, chymostatin treatment showed no 3D-RI changes and caused elevated fluctuations solely within the parasitophorous vacuole. We show that E64d and EGTA-AM supported PV breakdown and the resulting elevated fluctuations followed non-Gaussian pattern that resulted from direct merozoite impingement against the iRBC membrane. Optical trapping experiments highlighted reduced deformability of the iRBC membranes upon rupture-arrest, more specifically in the treatments that facilitated PV breakdown. Taken together, our experiments provide novel mechanistic interpretations on the role of parasitophorous vacuole in maintaining the spherical schizont morphology, the impact of PV breakdown on iRBC membrane fluctuations leading to eventual parasite escape and the evolution of membrane stiffness properties of host cells in which merozoites were irreversibly trapped, recourse to protease inhibitors. These findings provide a comprehensive, previously unavailable, body of information on the combined effects of biochemical and biophysical factors on parasite egress from iRBCs.

Porous Anodic Aluminum Oxide with Serrated Nanochannels

NASA Astrophysics Data System (ADS)

Li, Dongdong; Zhao, Liang; Lu, Jia G.

2010-03-01

Self-assembled nanoporous anodic aluminum oxide (AAO) membrane with straight channels has long been an important tool in synthesizing highly ordered and vertically aligned quasi-1D nanostructures for various applications. Recently shape-selective nanomaterials have been achieved using AAO as a template. It is envisioned that nanowires with multi-branches will significantly increase the active functional sites for applications as sensors, catalysts, chemical cells, etc. Here AAO membranes with serrated nanochannels have been successfully fabricated via a two-step annodization method. The serrated channels with periodic intervals are aligned at an angle of ˜25^circ along the stem channels. The formation of the serrated channels is attributed to the evolution of oxygen gas bubbles and the resulted plastic deformation in oxide membrane. In order to reveal the inside channel structure, Platinum are electrodeposited into the AAO template. The as-synthesized serrated Pt nanowires demonstrate a superior electrocatalytic activity. This is attributed to the enhanced electric field strength around serrated tips as shown in the electric field simulation by COMOSL. Moreover, hierarchical serrated/straight hybrid structures can be constructed using this simple and novel self assembly technique.

Membrane interactions between secretion granules and plasmalemma in three exocrine glands

PubMed Central

Tanaka, Y; De Camilli, P; Meldolesi, J

1980-01-01

Three types of membrane interactions were studied in three exocrine systems (the acinar cells of the rat parotid, rat lacrimal gland, and guinea pig pancrease) by freeze- fracture and thin-section electron microscopy: exocytosis, induced in vivo by specific pharmacological stimulations; the mutual apposition of secretory granule membranes in the intact cell; membrane appositions induced in vitro by centrifugation of the isolated granules. In all three glandular cells, the distribution of intramembrane particles (IMP) on the fracture faces of the luminal plasmagranule membrane particles (IMP) on the fracture faces of the lumenal plasmalemma appeared random before stimulation. However, after injection of secretagogues, IMP were rapidly clearly from the areas of granule- plasmalemma apposition in the parotid cells and, especially, in lacrimocytes. In the latter, the cleared areas appeared as large bulges toward the lumen, whereas in the parotid they were less pronounced. Exocytotic openings were usually large and the fracture faces of their rims were covered with IMP. In contrast, in stimulated pancreatic acinar cells, the IMP distribution remained apparently random after stimulation. Exocytoses were established through the formation of narrown necks, and no images which might correspond to early stages of membrane fusion were revealed. Within the cytoplasm of parotid and lacrimal cells (but not in the pancreas), both at rest and after stimulation, secretion granules were often closely apposed by means of flat, circular areas, also devoid of IMP. In thin sections, the images corresponding to IMP-free areas were close granule-granule and granule-plasmalemma appositions, sometimes with focal merging of the membrane outer layers to yield pentalaminar structures. Isolated secretion granules were forced together in vitro by centrifugation. Under these conditions, increasing the centrifugal force from 1,600 to 50,000 g for 10 min resulted in a progressive, statistically significant increase of the frequency of IMP-free flat appositions between parotid granules. In contrast, no such areas were seen between freeze-fractured pancreatic granules, although some focal pentalaminar appositions appeared in section after centrifugation at 50 and 100,000 g for 10 min. On the basis of the observation that, in secretory cells, IMP clearing always develops in deformed membrane areas (bulges, depressions, flat areas), it is suggested that it might result from the forced mechanical apposition of the interacting membranes. This might be a preliminary process not sufficient to initiate fusion. In the pancreas, IMP clearing could occur over surface areas too small to be detected. In stimulated parotid and lacrimal glands they were exceptional. These structures were either attached at the sites of continuity between granule and plasma membranes, or free in the acinar lumen, with a preferential location within exocytotic pockets or in their proximity. Experiments designed to investigate the nature of these blisters and vesicles revealed that they probably arise artifactually during glutaraldehyde fixation. In fact, (a) they were large and numerous in poorly fixed samples but were never observed in thin sections of specimens fixed in one step with glutaraldehyde and OsO(4); and (b) no increase in concentration of phospholipids was observed in the parotid saliva and pancreatic juice after stimulation of protein discharge, as was to be expected if release of membrane material were occurring after exocytosis. PMID:7380885

Deletion of the Vaccinia Virus I2 Protein Interrupts Virion Morphogenesis, Leading to Retention of the Scaffold Protein and Mislocalization of Membrane-Associated Entry Proteins.

PubMed

Hyun, Seong-In; Weisberg, Andrea; Moss, Bernard

2017-08-01

The I2L open reading frame of vaccinia virus (VACV) encodes a conserved 72-amino-acid protein with a putative C-terminal transmembrane domain. Previous studies with a tetracycline-inducible mutant demonstrated that I2-deficient virions are defective in cell entry. The purpose of the present study was to determine the step of replication or entry that is affected by loss of the I2 protein. Fluorescence microscopy experiments showed that I2 colocalized with a major membrane protein of immature and mature virions. We generated a cell line that constitutively expressed I2 and allowed construction of the VACV I2L deletion mutant vΔI2. As anticipated, vΔI2 was unable to replicate in cells that did not express I2. Unexpectedly, morphogenesis was interrupted at a stage after immature virion formation, resulting in the accumulation of dense spherical particles instead of brick-shaped mature virions with well-defined core structures. The abnormal particles retained the D13 scaffold protein of immature virions, were severely deficient in the transmembrane proteins that comprise the entry fusion complex (EFC), and had increased amounts of unprocessed membrane and core proteins. Total lysates of cells infected with vΔI2 also had diminished EFC proteins due to instability attributed to their hydrophobicity and failure to be inserted into viral membranes. A similar instability of EFC proteins had previously been found with unrelated mutants blocked earlier in morphogenesis that also accumulated viral membranes retaining the D13 scaffold. We concluded that I2 is required for virion morphogenesis, release of the D13 scaffold, and the association of EFC proteins with viral membranes. IMPORTANCE Poxviruses comprise a large family that infect vertebrates and invertebrates, cause disease in both in humans and in wild and domesticated animals, and are being engineered as vectors for vaccines and cancer therapy. In addition, investigations of poxviruses have provided insights into many aspects of cell biology. The I2 protein is conserved in all poxviruses that infect vertebrates, suggesting an important role. The present study revealed that this protein is essential for vaccinia virus morphogenesis and that its absence results in an accumulation of deformed virus particles retaining the scaffold protein and deficient in surface proteins needed for cell entry. Copyright © 2017 American Society for Microbiology.

Structure-based membrane dome mechanism for Piezo mechanosensitivity.

PubMed

Guo, Yusong R; MacKinnon, Roderick

2017-12-12

Mechanosensitive ion channels convert external mechanical stimuli into electrochemical signals for critical processes including touch sensation, balance, and cardiovascular regulation. The best understood mechanosensitive channel, MscL, opens a wide pore, which accounts for mechanosensitive gating due to in-plane area expansion. Eukaryotic Piezo channels have a narrow pore and therefore must capture mechanical forces to control gating in another way. We present a cryo-EM structure of mouse Piezo1 in a closed conformation at 3.7Å-resolution. The channel is a triskelion with arms consisting of repeated arrays of 4-TM structural units surrounding a pore. Its shape deforms the membrane locally into a dome. We present a hypothesis in which the membrane deformation changes upon channel opening. Quantitatively, membrane tension will alter gating energetics in proportion to the change in projected area under the dome. This mechanism can account for highly sensitive mechanical gating in the setting of a narrow, cation-selective pore. © 2017, Guo et al.

Flexoelectricity in two-dimensional crystalline and biological membranes

NASA Astrophysics Data System (ADS)

Ahmadpoor, Fatemeh; Sharma, Pradeep

2015-10-01

The ability of a material to convert electrical stimuli into mechanical deformation, i.e. piezoelectricity, is a remarkable property of a rather small subset of insulating materials. The phenomenon of flexoelectricity, on the other hand, is universal. All dielectrics exhibit the flexoelectric effect whereby non-uniform strain (or strain gradients) can polarize the material and conversely non-uniform electric fields may cause mechanical deformation. The flexoelectric effect is strongly enhanced at the nanoscale and accordingly, all two-dimensional membranes of atomistic scale thickness exhibit a strong two-way coupling between the curvature and electric field. In this review, we highlight the recent advances made in our understanding of flexoelectricity in two-dimensional (2D) membranes--whether the crystalline ones such as dielectric graphene nanoribbons or the soft lipid bilayer membranes that are ubiquitous in biology. Aside from the fundamental mechanisms, phenomenology, and recent findings, we focus on rapidly emerging directions in this field and discuss applications such as energy harvesting, understanding of the mammalian hearing mechanism and ion transport among others.

The ESCRT-III pathway facilitates cardiomyocyte release of cBIN1-containing microparticles

PubMed Central

Xu, Bing; Fu, Ying; Liu, Yan; Agvanian, Sosse; Wirka, Robert C.; Baum, Rachel; Zhou, Kang; Shaw, Robin M.

2017-01-01

Microparticles (MPs) are cell–cell communication vesicles derived from the cell surface plasma membrane, although they are not known to originate from cardiac ventricular muscle. In ventricular cardiomyocytes, the membrane deformation protein cardiac bridging integrator 1 (cBIN1 or BIN1+13+17) creates transverse-tubule (t-tubule) membrane microfolds, which facilitate ion channel trafficking and modulate local ionic concentrations. The microfold-generated microdomains continuously reorganize, adapting in response to stress to modulate the calcium signaling apparatus. We explored the possibility that cBIN1-microfolds are externally released from cardiomyocytes. Using electron microscopy imaging with immunogold labeling, we found in mouse plasma that cBIN1 exists in membrane vesicles about 200 nm in size, which is consistent with the size of MPs. In mice with cardiac-specific heterozygous Bin1 deletion, flow cytometry identified 47% less cBIN1-MPs in plasma, supporting cardiac origin. Cardiac release was also evidenced by the detection of cBIN1-MPs in medium bathing a pure population of isolated adult mouse cardiomyocytes. In human plasma, osmotic shock increased cBIN1 detection by enzyme-linked immunosorbent assay (ELISA), and cBIN1 level decreased in humans with heart failure, a condition with reduced cardiac muscle cBIN1, both of which support cBIN1 release in MPs from human hearts. Exploring putative mechanisms of MP release, we found that the membrane fission complex endosomal sorting complexes required for transport (ESCRT)-III subunit charged multivesicular body protein 4B (CHMP4B) colocalizes and coimmunoprecipitates with cBIN1, an interaction enhanced by actin stabilization. In HeLa cells with cBIN1 overexpression, knockdown of CHMP4B reduced the release of cBIN1-MPs. Using truncation mutants, we identified that the N-terminal BAR (N-BAR) domain in cBIN1 is required for CHMP4B binding and MP release. This study links the BAR protein superfamily to the ESCRT pathway for MP biogenesis in mammalian cardiac ventricular cells, identifying elements of a pathway by which cytoplasmic cBIN1 is released into blood. PMID:28806752

Effect of reconstructive vascular surgery on red cell deformability--preliminary results.

PubMed Central

Irwin, S T; Rocks, M J; McGuigan, J A; Patterson, C C; Morris, T C; O'Reilly, M J

1983-01-01

Using a simple filtration method, red cell deformability was measured in healthy control subjects and in patients with peripheral vascular disease. Impaired red cell deformability was demonstrated in patients with rest pain or gangrene and in patients with intermittent claudication. An improvement in red cell deformability was demonstrated after successful reconstructive vascular surgery in both patient groups. An improvement in red cell deformability was demonstrated in patients undergoing major limb amputation. PMID:6619311

Parametric Studies of Square Solar Sails Using Finite Element Analysis

NASA Technical Reports Server (NTRS)

Sleight, David W.; Muheim, Danniella M.

2004-01-01

Parametric studies are performed on two generic square solar sail designs to identify parameters of interest. The studies are performed on systems-level models of full-scale solar sails, and include geometric nonlinearity and inertia relief, and use a Newton-Raphson scheme to apply sail pre-tensioning and solar pressure. Computational strategies and difficulties encountered during the analyses are also addressed. The purpose of this paper is not to compare the benefits of one sail design over the other. Instead, the results of the parametric studies may be used to identify general response trends, and areas of potential nonlinear structural interactions for future studies. The effects of sail size, sail membrane pre-stress, sail membrane thickness, and boom stiffness on the sail membrane and boom deformations, boom loads, and vibration frequencies are studied. Over the range of parameters studied, the maximum sail deflection and boom deformations are a nonlinear function of the sail properties. In general, the vibration frequencies and modes are closely spaced. For some vibration mode shapes, local deformation patterns that dominate the response are identified. These localized patterns are attributed to the presence of negative stresses in the sail membrane that are artifacts of the assumption of ignoring the effects of wrinkling in the modeling process, and are not believed to be physically meaningful. Over the range of parameters studied, several regions of potential nonlinear modal interaction are identified.

Two-dimensional numerical modeling for separation of deformable cells using dielectrophoresis.

PubMed

Ye, Ting; Li, Hua; Lam, K Y

2015-02-01

In this paper, we numerically explore the possibility of separating two groups of deformable cells, by a very small dielectrophoretic (DEP) microchip with the characteristic length of several cell diameters. A 2D two-fluid model is developed to describe the separation process, where three types of forces are considered, the aggregation force for cell-cell interaction, the deformation force for cell deformation, and the DEP force for cell dielectrophoresis. As a model validation, we calculate the levitation height of a cell subject to DEP force, and compare it with the experimental data. After that, we simulate the separation of two groups of cells with different dielectric properties at high and low frequencies, respectively. The simulation results show that the deformable cells can be separated successfully by a very small DEP microchip, according to not only their different permittivities at the high frequency, but also their different conductivities at the low frequency. In addition, both two groups of cells have a shape deformation from an original shape to a lopsided slipper shape during the separation process. It is found that the cell motion is mainly determined by the DEP force arising from the electric field, causing the cells to deviate from the centerline of microchannel. However, the cell deformation is mainly determined by the deformation force arising from the fluid flow, causing the deviated cells to undergo an asymmetric motion with the deformation of slipper shape. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Dynamics of a bilayer membrane coupled to a two-dimensional cytoskeleton: Scale transfers of membrane deformations

NASA Astrophysics Data System (ADS)

Okamoto, Ryuichi; Komura, Shigeyuki; Fournier, Jean-Baptiste

2017-07-01

We theoretically investigate the dynamics of a floating lipid bilayer membrane coupled with a two-dimensional cytoskeleton network, taking into account explicitly the intermonolayer friction, the discrete lattice structure of the cytoskeleton, and its prestress. The lattice structure breaks lateral continuous translational symmetry and couples Fourier modes with different wave vectors. It is shown that within a short time interval a long-wavelength deformation excites a collection of modes with wavelengths shorter than the lattice spacing. These modes relax slowly with a common renormalized rate originating from the long-wavelength mode. As a result, and because of the prestress, the slowest relaxation is governed by the intermonolayer friction. Conversely, and most interestingly, forces applied at the scale of the cytoskeleton for a sufficiently long time can cooperatively excite large-scale modes.

Individually programmable cell stretching microwell arrays actuated by a Braille display.

PubMed

Kamotani, Yoko; Bersano-Begey, Tommaso; Kato, Nobuhiro; Tung, Yi-Chung; Huh, Dongeun; Song, Jonathan W; Takayama, Shuichi

2008-06-01

Cell culture systems are often static and are therefore nonphysiological. In vivo, many cells are exposed to dynamic surroundings that stimulate cellular responses in a process known as mechanotransduction. To recreate this environment, stretchable cell culture substrate systems have been developed, however, these systems are limited by being macroscopic and low throughput. We have developed a device consisting of 24 miniature cell stretching chambers with flexible bottom membranes that are deformed using the computer-controlled, piezoelectrically actuated pins of a Braille display. We have also developed efficient image capture and analysis protocols to quantify morphological responses of the cells to applied strain. Human dermal microvascular endothelial cells (HDMECs) were found to show increasing degrees of alignment and elongation perpendicular to the radial strain in response to cyclic stretch at increasing frequencies of 0.2, 1, and 5 Hz, after 2, 4, and 12h. Mouse myogenic C2C12 cells were also found to align in response to the stretch, while A549 human lung adenocarcinoma epithelial cells did not respond to stretch.

Individually Programmable Cell Stretching Microwell Arrays Actuated by a Braille Display

PubMed Central

Kamotani, Yoko; Bersano-Begey, Tommaso; Kato, Nobuhiro; Tung, Yi-chung; Huh, Dongeun; Song, Jonathan W.; Takayama, Shuichi

2008-01-01

Cell culture systems are often static and are therefore nonphysiological. In vivo, many cells are exposed to dynamic surroundings that stimulate cellular responses in a process known as mechanotransduction. To recreate this environment, stretchable cell culture substrate systems have been developed, however these systems are limited by being macroscopic and low throughput. We have developed a device consisting of 24 miniature cell stretching chambers with flexible bottom membranes that are deformed using the computer-controlled, piezoelectrically actuated pins of a Braille display. We have also developed efficient image capture and analysis protocols to quantify morphological responses of the cells to applied strain. Human dermal microvascular endothelial cells (HDMECs) were found to show increasing degrees of alignment and elongation perpendicular to the radial strain in response to cyclic stretch at increasing frequencies of 0.2, 1, and 5 Hz, after 2, 4, and 12 hours. Mouse myogenic C2C12 cells were also found to align in response to the stretch, while A549 human lung adenocarcinoma epithelial cells did not respond to stretch. PMID:18342367

Stretchable Biofuel Cells as Wearable Textile-based Self-Powered Sensors.

PubMed

Jeerapan, Itthipon; Sempionatto, Juliane R; Pavinatto, Adriana; You, Jung-Min; Wang, Joseph

2016-12-21

Highly stretchable textile-based biofuel cells (BFCs), acting as effective self-powered sensors, have been fabricated using screen-printing of customized stress-enduring inks. Due to synergistic effects of nanomaterial-based engineered inks and the serpentine designs, these printable bioelectronic devices endure severe mechanical deformations, e.g., stretching, indentation, or torsional twisting. Glucose and lactate BFCs with the single enzyme and membrane-free configurations generated the maximum power density of 160 and 250 µW cm -2 with the open circuit voltages of 0.44 and 0.46 V, respectively. The textile-BFCs were able to withstand repeated severe mechanical deformations with minimal impact on its structural integrity, as was indicated from their stable power output after 100 cycles of 100% stretching. By providing power signals proportional to the sweat fuel concentration, these stretchable devices act as highly selective and stable self-powered textile sensors. Applicability to sock-based BFC and self-powered biosensor and mechanically compliant operations was demonstrated on human subjects. These stretchable skin-worn "scavenge-sense-display" devices are expected to contribute to the development of skin-worn energy harvesting systems, advanced non-invasive self-powered sensors and wearable electronics on a stretchable garment.

Seismic anisotropy of the D'' layer induced by (001) deformation of post-perovskite

DOE Office of Scientific and Technical Information (OSTI.GOV)

Wu, Xiang; Lin, Jung-Fu; Kaercher, Pamela

Crystallographic preferred orientation (CPO) of post-perovskite (Mg,Fe)SiO 3 (pPv) has been believed to be one potential source of the seismic anisotropic layer at the bottom of the lower mantle (D'' layer). However, the natural CPO of pPv remains ambiguous in the D'' layer. Here we have carried out the deformation experiments of pPv-(Mg 0.75,Fe 0.25)SiO 3 using synchrotron radial X-ray diffraction in a membrane-driven laser-heated diamond anvil cell from 135 GPa and 2,500 K to 154 GPa and 3,000 K. Our results show that the intrinsic texture of pPv-(Mg 0.75,Fe 0.25)SiO 3 should be (001) at realistic P–T conditions ofmore » the D'' layer, which can produce a shear wave splitting anisotropy of ~3.7% with V SH>V SV. Considering the combined effect of both pPv and ferropericlase, we suggest that 50% or less of deformation is sufficient to explain the origin of the shear wave anisotropy observed seismically in the D'' layer beneath the circum-Pacific rim.« less

Full Eulerian simulations of biconcave neo-Hookean particles in a Poiseuille flow

NASA Astrophysics Data System (ADS)

Sugiyama, Kazuyasu; , Satoshi, II; Takeuchi, Shintaro; Takagi, Shu; Matsumoto, Yoichiro

2010-03-01

For a given initial configuration of a multi-component geometry represented by voxel-based data on a fixed Cartesian mesh, a full Eulerian finite difference method facilitates solution of dynamic interaction problems between Newtonian fluid and hyperelastic material. The solid volume fraction, and the left Cauchy-Green deformation tensor are temporally updated on the Eulerian frame, respectively, to distinguish the fluid and solid phases, and to describe the solid deformation. The simulation method is applied to two- and three-dimensional motions of two biconcave neo-Hookean particles in a Poiseuille flow. Similar to the numerical study on the red blood cell motion in a circular pipe (Gong et al. in J Biomech Eng 131:074504, 2009), in which Skalak’s constitutive laws of the membrane are considered, the deformation, the relative position and orientation of a pair of particles are strongly dependent upon the initial configuration. The increase in the apparent viscosity is dependent upon the developed arrangement of the particles. The present Eulerian approach is demonstrated that it has the potential to be easily extended to larger system problems involving a large number of particles of complicated geometries.

Origins of chemoreceptor curvature sorting in Escherichia coli

PubMed Central

Draper, Will; Liphardt, Jan

2017-01-01

Bacterial chemoreceptors organize into large clusters at the cell poles. Despite a wealth of structural and biochemical information on the system's components, it is not clear how chemoreceptor clusters are reliably targeted to the cell pole. Here, we quantify the curvature-dependent localization of chemoreceptors in live cells by artificially deforming growing cells of Escherichia coli in curved agar microchambers, and find that chemoreceptor cluster localization is highly sensitive to membrane curvature. Through analysis of multiple mutants, we conclude that curvature sensitivity is intrinsic to chemoreceptor trimers-of-dimers, and results from conformational entropy within the trimer-of-dimers geometry. We use the principles of the conformational entropy model to engineer curvature sensitivity into a series of multi-component synthetic protein complexes. When expressed in E. coli, the synthetic complexes form large polar clusters, and a complex with inverted geometry avoids the cell poles. This demonstrates the successful rational design of both polar and anti-polar clustering, and provides a synthetic platform on which to build new systems. PMID:28322223

Neuronize: a tool for building realistic neuronal cell morphologies

PubMed Central

Brito, Juan P.; Mata, Susana; Bayona, Sofia; Pastor, Luis; DeFelipe, Javier; Benavides-Piccione, Ruth

2013-01-01

This study presents a tool, Neuronize, for building realistic three-dimensional models of neuronal cells from the morphological information extracted through computer-aided tracing applications. Neuronize consists of a set of methods designed to build 3D neural meshes that approximate the cell membrane at different resolution levels, allowing a balance to be reached between the complexity and the quality of the final model. The main contribution of the present study is the proposal of a novel approach to build a realistic and accurate 3D shape of the soma from the incomplete information stored in the digitally traced neuron, which usually consists of a 2D cell body contour. This technique is based on the deformation of an initial shape driven by the position and thickness of the first order dendrites. The addition of a set of spines along the dendrites completes the model, building a final 3D neuronal cell suitable for its visualization in a wide range of 3D environments. PMID:23761740

Neuronize: a tool for building realistic neuronal cell morphologies.

PubMed

Brito, Juan P; Mata, Susana; Bayona, Sofia; Pastor, Luis; Defelipe, Javier; Benavides-Piccione, Ruth

2013-01-01

This study presents a tool, Neuronize, for building realistic three-dimensional models of neuronal cells from the morphological information extracted through computer-aided tracing applications. Neuronize consists of a set of methods designed to build 3D neural meshes that approximate the cell membrane at different resolution levels, allowing a balance to be reached between the complexity and the quality of the final model. The main contribution of the present study is the proposal of a novel approach to build a realistic and accurate 3D shape of the soma from the incomplete information stored in the digitally traced neuron, which usually consists of a 2D cell body contour. This technique is based on the deformation of an initial shape driven by the position and thickness of the first order dendrites. The addition of a set of spines along the dendrites completes the model, building a final 3D neuronal cell suitable for its visualization in a wide range of 3D environments.

Measuring skewness of red blood cell deformability distribution by laser ektacytometry

DOE Office of Scientific and Technical Information (OSTI.GOV)

Nikitin, S Yu; Priezzhev, A V; Lugovtsov, A E

An algorithm is proposed for measuring the parameters of red blood cell deformability distribution based on laser diffractometry of red blood cells in shear flow (ektacytometry). The algorithm is tested on specially prepared samples of rat blood. In these experiments we succeeded in measuring the mean deformability, deformability variance and skewness of red blood cell deformability distribution with errors of 10%, 15% and 35%, respectively. (laser biophotonics)

Simulation of controllable permeation in PNIPAAm coated membranes

NASA Astrophysics Data System (ADS)

Ehrenhofer, Adrian; Wallmersperger, Thomas; Richter, Andreas

2016-04-01

Membranes separate fluid compartments and can comprise transport structures for selective permeation. In biology, channel proteins are specialized in their atomic structure to allow transport of specific compounds (selectivity). Conformational changes in protein structure allow the control of the permeation abilities by outer stimuli (gating). In polymeric membranes, the selectivity is due to electrostatic or size-exclusion. It can thus be controlled by size variation or electric charges. Controllable permeation can be useful to determine particle-size distributions in continuous flow, e.g. in microfluidics and biomedicine to gain cell diameter profiles in blood. The present approach uses patterned polyethylene terephthalate (PET) membranes with hydrogel surface coating for permeation control by size-exclusion. The thermosensitive hydrogel poly(N-isopropylacrylamide) (PNIPAAm) is structured with a cross-shaped pore geometry. A change in the temperature of the water flow through the membrane leads to a pore shape variation. The temperature dependent behavior of PNIPAAm can be numerically modeled with a temperature expansion model, where the swelling and deswelling is depicted by temperature dependent expansion coefficients. In the present study, the free swelling behavior was implemented to the Finite Element tool ABAQUS for the complex composite structure of the permeation control membrane. Experimental values of the geometry characteristics were derived from microscopy images with the tool Image J and compared to simulation results. Numerical simulations using the derived thermo-mechanical model for different pore geometries (circular, rectangle, cross and triangle) were performed. With this study, we show that the temperature expansion model with values from the free swelling behavior can be used to adequately predict the deformation behavior of the complex membrane system. The predictions can be used to optimize the behavior of the membrane pores and the overall performance of the smart membrane.

Effect of electromagnetic fields on some biomechanical and biochemical properties of rat’s blood

NASA Astrophysics Data System (ADS)

Mohaseb, M. A.; Shahin, F. A.; Ali, F. M.; Baieth, H. A.

2017-06-01

In order to study the effect of electromagnetic fields (0.3 mT, 50 Hz) on some biomechanical and biochemical properties of rats’ blood, healthy thirty male albino rats of 150 ± 10 g were divided into three equal groups namely A, B1, B2. Group A used as a control group, group B1 was continuously exposed to a magnetic field of (0.3 mT, 50 Hz) for a period of 21 days for direct effect studies. Group B2 was continuously exposed to the same magnetic field for the same period of time, then was housed away from the magnetic field for a period of 45 days for delayed effects studies. After examination, the results indicated that the apparent viscosity and the consistency index increased significantly and very high significantly for groub B1 and B2 compared to control at P<0.05. Red blood cell counts (RBCs) membrane elasticity had significantly and very high significantly decreased for groups B1 and B2. Moreover, delayed effects studies indicated that there is deterioration in the bone marrow functions. These results are supported by the blood film image, where irregularities and deformations in the RBCs membranes had been occurred. We conclude that the cell membrane properties are highly affected by the extremely low frequency (ELF) magnetic fields, which proved to be biologically toxic.

Three-dimensional multi-scale model of deformable platelets adhesion to vessel wall in blood flow

PubMed Central

Wu, Ziheng; Xu, Zhiliang; Kim, Oleg; Alber, Mark

2014-01-01

When a blood vessel ruptures or gets inflamed, the human body responds by rapidly forming a clot to restrict the loss of blood. Platelets aggregation at the injury site of the blood vessel occurring via platelet–platelet adhesion, tethering and rolling on the injured endothelium is a critical initial step in blood clot formation. A novel three-dimensional multi-scale model is introduced and used in this paper to simulate receptor-mediated adhesion of deformable platelets at the site of vascular injury under different shear rates of blood flow. The novelty of the model is based on a new approach of coupling submodels at three biological scales crucial for the early clot formation: novel hybrid cell membrane submodel to represent physiological elastic properties of a platelet, stochastic receptor–ligand binding submodel to describe cell adhesion kinetics and lattice Boltzmann submodel for simulating blood flow. The model implementation on the GPU cluster significantly improved simulation performance. Predictive model simulations revealed that platelet deformation, interactions between platelets in the vicinity of the vessel wall as well as the number of functional GPIbα platelet receptors played significant roles in platelet adhesion to the injury site. Variation of the number of functional GPIbα platelet receptors as well as changes of platelet stiffness can represent effects of specific drugs reducing or enhancing platelet activity. Therefore, predictive simulations can improve the search for new drug targets and help to make treatment of thrombosis patient-specific. PMID:24982253

Analyses of quasi-isotropic composite plates under quasi-static point loads simulating low-velocity impact phenomena

NASA Technical Reports Server (NTRS)

Kelkar, A. D.

1984-01-01

In thin composite laminates, the first level of visible damage occurs in the back face and is called back face spalling. A plate-membrane coupling model, and a finite element model to analyze the large deformation behavior of eight-ply quasi-isotropic circular composite plates under impact type point loads are developed. The back face spalling phenomenon in thin composite plates is explained by using the plate-membrane coupling model and the finite element model in conjunction with the fracture mechanics principles. The experimental results verifying these models are presented. Several conclusions concerning the deformation behavior are reached and discussed in detail.

Failure mechanism of monolayer graphene under hypervelocity impact of spherical projectile

PubMed Central

Xia, Kang; Zhan, Haifei; Hu, De’an; Gu, Yuantong

2016-01-01

The excellent mechanical properties of graphene have enabled it as appealing candidate in the field of impact protection or protective shield. By considering a monolayer graphene membrane, in this work, we assessed its deformation mechanisms under hypervelocity impact (from 2 to 6 km/s), based on a serial of in silico studies. It is found that the cracks are formed preferentially in the zigzag directions which are consistent with that observed from tensile deformation. Specifically, the boundary condition is found to exert an obvious influence on the stress distribution and transmission during the impact process, which eventually influences the penetration energy and crack growth. For similar sample size, the circular shape graphene possesses the best impact resistance, followed by hexagonal graphene membrane. Moreover, it is found the failure shape of graphene membrane has a strong relationship with the initial kinetic energy of the projectile. The higher kinetic energy, the more number the cracks. This study provides a fundamental understanding of the deformation mechanisms of monolayer graphene under impact, which is crucial in order to facilitate their emerging future applications for impact protection, such as protective shield from orbital debris for spacecraft. PMID:27618989

Failure mechanism of monolayer graphene under hypervelocity impact of spherical projectile

NASA Astrophysics Data System (ADS)

Xia, Kang; Zhan, Haifei; Hu, De'An; Gu, Yuantong

2016-09-01

The excellent mechanical properties of graphene have enabled it as appealing candidate in the field of impact protection or protective shield. By considering a monolayer graphene membrane, in this work, we assessed its deformation mechanisms under hypervelocity impact (from 2 to 6 km/s), based on a serial of in silico studies. It is found that the cracks are formed preferentially in the zigzag directions which are consistent with that observed from tensile deformation. Specifically, the boundary condition is found to exert an obvious influence on the stress distribution and transmission during the impact process, which eventually influences the penetration energy and crack growth. For similar sample size, the circular shape graphene possesses the best impact resistance, followed by hexagonal graphene membrane. Moreover, it is found the failure shape of graphene membrane has a strong relationship with the initial kinetic energy of the projectile. The higher kinetic energy, the more number the cracks. This study provides a fundamental understanding of the deformation mechanisms of monolayer graphene under impact, which is crucial in order to facilitate their emerging future applications for impact protection, such as protective shield from orbital debris for spacecraft.

Vesicle deformation by microtubules: A phase diagram

NASA Astrophysics Data System (ADS)

Emsellem, Virginie; Cardoso, Olivier; Tabeling, Patrick

1998-10-01

The experimental investigation of vesicles deformed by the growth of encapsulated microtubules shows that the axisymmetric morphologies can be classified into ovals, lemons, φ, cherries, dumbbells, and pearls. A geometrical phase diagram is established. Numerical minimization of the elastic energy of the membrane reproduces satisfactorily well the observed morphologies and the corresponding phase diagram.

Modeling Membrane Deformations and Lipid Demixing upon Protein-Membrane Interaction: The BAR Dimer Adsorption

PubMed Central

Khelashvili, George; Harries, Daniel; Weinstein, Harel

2009-01-01

We use a self-consistent mean-field theory, designed to investigate membrane reshaping and lipid demixing upon interaction with proteins, to explore BAR domains interacting with large patches of lipid membranes of heterogeneous compositions. The computational model includes contributions to the system free energy from electrostatic interactions and elastic energies of the membrane, as well as salt and lipid mixing entropies. The results from our simulation of a single adsorbing Amphiphysin BAR dimer indicate that it is capable of stabilizing a significantly curved membrane. However, we predict that such deformations will occur only for membrane patches that have the inherent propensity for high curvature, reflected in the tendency to create local distortions that closely match the curvature of the BAR dimer itself. Such favorable preconditioning for BAR-membrane interaction may be the result of perturbations such as local lipid demixing induced by the interaction, or of a prior insertion of the BAR domain's amphiphatic N-helix. From our simulations it appears that local segregation of charged lipids under the influence of the BAR dimer cannot produce high enough asymmetry between bilayer leaflets to induce significant bending. In the absence of additional energy contributions that favor membrane asymmetry, the membrane will remain nearly flat upon single BAR dimer adsorption, relative to the undulation expected from thermal fluctuations. Thus, we conclude that the N-helix insertions have a critical mechanistic role in the local perturbation and curving of the membrane, which is then stabilized by the electrostatic interaction with the BAR dimer. We discuss how these results can be used to estimate the tendency of BARs to bend membranes in terms of a spatially nonisotropic spontaneous curvature. PMID:19751667

Modeling and Analysis of Wrinkled Membranes: An Overview

NASA Technical Reports Server (NTRS)

Yang, B.; Ding, H.; Lou, M.; Fang, H.; Broduer, Steve (Technical Monitor)

2001-01-01

Thin-film membranes are basic elements of a variety of space inflatable/deployable structures. Wrinkling degrades the performance and reliability of these membrane structures, and hence has been a topic of continued interest. Wrinkling analysis of membranes for general geometry and arbitrary boundary conditions is quite challenging. The objective of this presentation is two-fold. Firstly, the existing models of wrinkled membranes and related numerical solution methods are reviewed. The important issues to be discussed are the capability of a membrane model to characterize taut, wrinkled and slack states of membranes in a consistent and physically reasonable manner; the ability of a wrinkling analysis method to predict the formation and growth of wrinkled regions, and to determine out-of-plane deformation and wrinkled waves; the convergence of a numerical solution method for wrinkling analysis; and the compatibility of a wrinkling analysis with general-purpose finite element codes. According to this review, several opening issues in modeling and analysis of wrinkled membranes that are to be addressed in future research are summarized, The second objective of this presentation is to discuss a newly developed membrane model of two viable parameters (2-VP model) and associated parametric finite element method (PFEM) for wrinkling analysis are introduced. The innovations and advantages of the proposed membrane model and PFEM-based wrinkling analysis are: (1) Via a unified stress-strain relation; the 2-VP model treat the taut, wrinkled, and slack states of membranes consistently; (2) The PFEM-based wrinkling analysis has guaranteed convergence; (3) The 2-VP model along with PFEM is capable of predicting membrane out-of-plane deformations; and (4) The PFEM can be integrated into any existing finite element code. Preliminary numerical examples are also included in this presentation to demonstrate the 2-VP model and PFEM-based wrinkling analysis approach.

4D metrology of flapping-wing micro air vehicle based on fringe projection

NASA Astrophysics Data System (ADS)

Zhang, Qican; Huang, Lei; Chin, Yao-Wei; Keong, Lau-Gih; Asundi, Anand

2013-06-01

Inspired by dominant flight of the natural flyers and driven by civilian and military purposes, micro air vehicle (MAV) has been developed so far by passive wing control but still pales in aerodynamic performance. Better understanding of flapping wing flight mechanism is eager to improve MAV's flight performance. In this paper, a simple and effective 4D metrology technique to measure full-field deformation of flapping membrane wing is presented. Based on fringe projection and 3D Fourier analysis, the fast and complex dynamic deformation, including wing rotation and wing stroke, of a flapping wing during its flight can be accurately reconstructed from the deformed fringe patterns recorded by a highspeed camera. An experiment was carried on a flapping-wing MAV with 5-cm span membrane wing beating at 30 Hz, and the results show that this method is effective and will be useful to the aerodynamicist or micro aircraft designer for visualizing high-speed complex wing deformation and consequently aid the design of flapping wing mechanism to enhanced aerodynamic performance.

Effect of bending on the dynamics and wrinkle formation for a capsule in shear flow

NASA Astrophysics Data System (ADS)

Salsac, Anne-Virginie; Dupont, Claire; Barthes-Biesel, Dominique; Vidrascu, Marina; Le Tallec, Patrick

2014-11-01

When microcapsules are subjected to an external flow, the droplets enclosed within a thin hyperelastic wall undergo large deformations, which often lead to buckling of the thin capsule wall. The objective is to study numerically an initially spherical capsule in shear flow and analyze the influence of the membrane bending rigidity on the capsule dynamics and wrinkle formation. The 3D fluid-structure interactions are modeled coupling a boundary integral method to solve for the internal and external Stokes flows with a thin shell finite element method to solve for the wall deformation. Hyperelastic constitutive laws are implemented to model the deformation of the capsule mid-surface and the generalized Hooke's law for the bending effects. We show that the capsule global motion and deformation are mainly governed by in-plane membrane tensions and are marginally influenced by the bending stiffness Ks. The bending stiffness, however, plays a role locally in regions of compressive tensions. The wrinkle wavelength depends on Ks following a power law, which provides an experimental technique to determine the value of Ks through inverse analysis.

Cancer cells become less deformable and more invasive with activation of β-adrenergic signaling

PubMed Central

Gill, Navjot Kaur; Nyberg, Kendra D.; Nguyen, Angelyn V.; Hohlbauch, Sophia V.; Geisse, Nicholas A.; Nowell, Cameron J.; Sloan, Erica K.

2016-01-01

ABSTRACT Invasion by cancer cells is a crucial step in metastasis. An oversimplified view in the literature is that cancer cells become more deformable as they become more invasive. β-adrenergic receptor (βAR) signaling drives invasion and metastasis, but the effects on cell deformability are not known. Here, we show that activation of β-adrenergic signaling by βAR agonists reduces the deformability of highly metastatic human breast cancer cells, and that these stiffer cells are more invasive in vitro. We find that βAR activation also reduces the deformability of ovarian, prostate, melanoma and leukemia cells. Mechanistically, we show that βAR-mediated cell stiffening depends on the actin cytoskeleton and myosin II activity. These changes in cell deformability can be prevented by pharmacological β-blockade or genetic knockout of the β2-adrenergic receptor. Our results identify a β2-adrenergic–Ca2+–actin axis as a new regulator of cell deformability, and suggest that the relationship between cell mechanical properties and invasion might be dependent on context. PMID:27875276

Immature HIV-1 lattice assembly dynamics are regulated by scaffolding from nucleic acid and the plasma membrane

PubMed Central

Pak, Alexander J.; Grime, John M. A.; Sengupta, Prabuddha; Chen, Antony K.; Durumeric, Aleksander E. P.; Srivastava, Anand; Yeager, Mark; Briggs, John A. G.; Lippincott-Schwartz, Jennifer; Voth, Gregory A.

2017-01-01

The packaging and budding of Gag polyprotein and viral RNA is a critical step in the HIV-1 life cycle. High-resolution structures of the Gag polyprotein have revealed that the capsid (CA) and spacer peptide 1 (SP1) domains contain important interfaces for Gag self-assembly. However, the molecular details of the multimerization process, especially in the presence of RNA and the cell membrane, have remained unclear. In this work, we investigate the mechanisms that work in concert between the polyproteins, RNA, and membrane to promote immature lattice growth. We develop a coarse-grained (CG) computational model that is derived from subnanometer resolution structural data. Our simulations recapitulate contiguous and hexameric lattice assembly driven only by weak anisotropic attractions at the helical CA–SP1 junction. Importantly, analysis from CG and single-particle tracking photoactivated localization (spt-PALM) trajectories indicates that viral RNA and the membrane are critical constituents that actively promote Gag multimerization through scaffolding, while overexpression of short competitor RNA can suppress assembly. We also find that the CA amino-terminal domain imparts intrinsic curvature to the Gag lattice. As a consequence, immature lattice growth appears to be coupled to the dynamics of spontaneous membrane deformation. Our findings elucidate a simple network of interactions that regulate the early stages of HIV-1 assembly and budding. PMID:29114055

Surface-Tension Replica-Exchange Molecular Dynamics Method for Enhanced Sampling of Biological Membrane Systems.

PubMed

Mori, Takaharu; Jung, Jaewoon; Sugita, Yuji

2013-12-10

Conformational sampling is fundamentally important for simulating complex biomolecular systems. The generalized-ensemble algorithm, especially the temperature replica-exchange molecular dynamics method (T-REMD), is one of the most powerful methods to explore structures of biomolecules such as proteins, nucleic acids, carbohydrates, and also of lipid membranes. T-REMD simulations have focused on soluble proteins rather than membrane proteins or lipid bilayers, because explicit membranes do not keep their structural integrity at high temperature. Here, we propose a new generalized-ensemble algorithm for membrane systems, which we call the surface-tension REMD method. Each replica is simulated in the NPγT ensemble, and surface tensions in a pair of replicas are exchanged at certain intervals to enhance conformational sampling of the target membrane system. We test the method on two biological membrane systems: a fully hydrated DPPC (1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine) lipid bilayer and a WALP23-POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) membrane system. During these simulations, a random walk in surface tension space is realized. Large-scale lateral deformation (shrinking and stretching) of the membranes takes place in all of the replicas without collapse of the lipid bilayer structure. There is accelerated lateral diffusion of DPPC lipid molecules compared with conventional MD simulation, and a much wider range of tilt angle of the WALP23 peptide is sampled due to large deformation of the POPC lipid bilayer and through peptide-lipid interactions. Our method could be applicable to a wide variety of biological membrane systems.

Actively tunable transverse waves in soft membrane-type acoustic metamaterials

NASA Astrophysics Data System (ADS)

Zhou, Weijian; Wu, Bin; Muhammad, Du, Qiujiao; Huang, Guoliang; Lü, Chaofeng; Chen, Weiqiu

2018-04-01

Membrane-type metamaterials have shown a fantastic capacity for manipulating acoustic waves in the low frequency range. They have the advantages of simple geometry, light weight, and active tunability. In general, these membrane-type metamaterials contain a rigid frame support, leading to a fixed configuration. However, in some instances, flexible and reconfigurable devices may be desirable. A soft membrane-type acoustic metamaterial that is highly flexible and controllable is designed here. Different from the previously designed membrane-type metamaterials, the stiff supporting frame is removed and the stiff mass at the center of each unit cell is replaced by the soft mass, realized by bonding fine metallic particles in the central region. In contrast to the previous studies, the propagation of elastic transverse waves in such a soft metamaterial is investigated by employing the plane wave expansion method. Both the Bragg scattering bandgaps and locally resonant bandgaps are found to coexist in the soft metamaterial. The influences of structural parameters and finite biaxial pre-stretch on the dynamic behavior of this soft metamaterial are carefully examined. It is shown that whether or not the wave propagation characteristics are sensitive to the finite deformation does not depend on the property and pre-stretch of the membrane. In addition, a broadband complete bandgap and a pseudo-gap formed by the combination of two extremely adjacent directional bandgaps are observed in the low-frequency range, and both can be controlled by the finite pre-stretch.

The Plasmodium falciparum exported protein PF3D7_0402000 binds to erythrocyte ankyrin and band 4.1

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shakya, Bikash; Penn, Wesley D.; Nakayasu, Ernesto S.

Plasmodium falciparum extensively modifies the infected red blood cell (RBC), resulting in changes in deformability, shape and surface properties. These alterations suggest that the RBC cytoskeleton is a major target for modification during infection. However, the molecular mechanisms leading to these changes are largely unknown. To begin to address this question, we screened for exported P. falciparum proteins that bound to the erythrocyte cytoskeleton proteins ankyrin 1 (ANK1) and band 4.1 (4.1R), which form critical interactions with other cytoskeletal proteins that contribute to the deformability and stability of RBCs. Yeast two-hybrid screens with ANK1 and 4.1R identified eight interactions withmore » P. falciparum exported proteins, including an interaction between 4.1R and PF3D7_0402000 (PFD0090c). This interaction was first identified in a large-scale screen (Vignali et al., Malaria J, 7:211, 2008), which also reported an interaction between PF3D7_0402000 and ANK1. We confirmed the interactions of PF3D7_0402000 with 4.1R and ANK1 in pair-wise yeast two-hybrid and co-precipitation assays. In both cases, an intact PHIST domain in PF3D7_0402000 was required for binding. Complex purification followed by mass spectrometry analysis provided additional support for the interaction of PF3D7_0402000 with ANK1 and 4.1R. RBC ghost cells loaded with maltose-binding protein (MBP)-PF3D7_0402000 passed through a metal microsphere column less efficiently than mock- or MBP-loaded controls, consistent with an effect of PF3D7_0402000 on RBC rigidity or membrane stability. This study confirmed the interaction of PF3D7_0402000 with 4.1R in multiple independent assays, provided the first evidence that PF3D7_0402000 also binds to ANK1, and suggested that PF3D7_0402000 affects deformability or membrane stability of uninfected RBC ghosts.« less

The effect and mechanism of inhibiting glucose-6-phosphate dehydrogenase activity on the proliferation of Plasmodium falciparum.

PubMed

Zhang, Zhiqiang; Chen, Xiaodan; Jiang, Chengrui; Fang, Zishui; Feng, Yi; Jiang, Weiying

2017-05-01

We screened >40,000 patients with glucose-6-phosphate dehydrogenase (G6PD) deficiency and found that the G6PD Kaiping allele was under the most positive selection for fighting against malaria in the Chinese population. However, the mechanism is unknown. The current study was designed to investigate the anti-malarial effect and mechanism of G6PD deficiency. Dehydroepiandrosterone (DHEA) was utilised for inhibiting the G6PD activity of erythrocytes. Giemsa staining of blood smears and quantitative real-time PCR were used for the detection and quantification of Plasmodium falciparum infection. A transmission electron microscope was used to observe the structural changes of P. falciparum. An atomic force microscopy was used for the analyses of morphology, roughness and Young's Modulus of the infective erythrocyte membrane. When G6PD activity was inhibited by DHEA, the infection rate of P. falciparum decreased, its cell nucleus shrank, the cell organelles and metabolites were reduced gradually and the Young's Modulus of the erythrocyte membrane increased with increasing DHEA concentrations. These data indicated that Plasmodium multiplication would be inhibited in G6PD deficient erythrocytes because the Plasmodium organelles could not obtain enough nutrients, including ribose-5-phosphate and the reducing equivalent, NADPH. Moreover, the Young's Modulus of the erythrocyte membrane increased, which resulted in an increased membrane stiffness and decreased deformation. It was difficult for the merozoites to invade erythrocytes through endocytosis. Understanding these points will have a major effect on searching for new anti-malarial drug targets. Copyright © 2017 Elsevier B.V. All rights reserved.

Calpain-1 knockout reveals broad effects on erythrocyte deformability and physiology

PubMed Central

Wieschhaus, Adam; Khan, Anwar; Zaidi, Asma; Rogalin, Henry; Hanada, Toshihiko; Liu, Fei; De Franceschi, Lucia; Brugnara, Carlo; Rivera, Alicia; Chishti, Athar H.

2014-01-01

Pharmacological inhibitors of cysteine proteases have provided useful insights into the regulation of calpain activity in erythrocytes. However, the precise biological function of calpain activity in erythrocytes remains poorly understood. Erythrocytes express calpain-1, an isoform regulated by calpastatin, the endogenous inhibitor of calpains. In the present study, we investigated the function of calpain-1 in mature erythrocytes using our calpain-1-null [KO (knockout)] mouse model. The calpain-1 gene deletion results in improved erythrocyte deformability without any measurable effect on erythrocyte lifespan in vivo. The calcium-induced sphero-echinocyte shape transition is compromised in the KO erythrocytes. Erythrocyte membrane proteins ankyrin, band 3, protein 4.1R, adducin and dematin are degraded in the calcium-loaded normal erythrocytes but not in the KO erythrocytes. In contrast, the integrity of spectrin and its state of phosphorylation are not affected in the calcium-loaded erythrocytes of either genotype. To assess the functional consequences of attenuated cytoskeletal remodelling in the KO erythrocytes, the activity of major membrane transporters was measured. The activity of the K+–Cl− co-transporter and the Gardos channel was significantly reduced in the KO erythrocytes. Similarly, the basal activity of the calcium pump was reduced in the absence of calmodulin in the KO erythrocyte membrane. Interestingly, the calmodulin-stimulated calcium pump activity was significantly elevated in the KO erythrocytes, implying a wider range of pump regulation by calcium and calmodulin. Taken together, and with the atomic force microscopy of the skeletal network, the results of the present study provide the first evidence for the physiological function of calpain-1 in erythrocytes with therapeutic implications for calcium imbalance pathologies such as sickle cell disease. PMID:22870887

Calpain-1 knockout reveals broad effects on erythrocyte deformability and physiology.

PubMed

Wieschhaus, Adam; Khan, Anwar; Zaidi, Asma; Rogalin, Henry; Hanada, Toshihiko; Liu, Fei; De Franceschi, Lucia; Brugnara, Carlo; Rivera, Alicia; Chishti, Athar H

2012-11-15

Pharmacological inhibitors of cysteine proteases have provided useful insights into the regulation of calpain activity in erythrocytes. However, the precise biological function of calpain activity in erythrocytes remains poorly understood. Erythrocytes express calpain-1, an isoform regulated by calpastatin, the endogenous inhibitor of calpains. In the present study, we investigated the function of calpain-1 in mature erythrocytes using our calpain-1-null [KO (knockout)] mouse model. The calpain-1 gene deletion results in improved erythrocyte deformability without any measurable effect on erythrocyte lifespan in vivo. The calcium-induced sphero-echinocyte shape transition is compromised in the KO erythrocytes. Erythrocyte membrane proteins ankyrin, band 3, protein 4.1R, adducin and dematin are degraded in the calcium-loaded normal erythrocytes but not in the KO erythrocytes. In contrast, the integrity of spectrin and its state of phosphorylation are not affected in the calcium-loaded erythrocytes of either genotype. To assess the functional consequences of attenuated cytoskeletal remodelling in the KO erythrocytes, the activity of major membrane transporters was measured. The activity of the K+-Cl- co-transporter and the Gardos channel was significantly reduced in the KO erythrocytes. Similarly, the basal activity of the calcium pump was reduced in the absence of calmodulin in the KO erythrocyte membrane. Interestingly, the calmodulin-stimulated calcium pump activity was significantly elevated in the KO erythrocytes, implying a wider range of pump regulation by calcium and calmodulin. Taken together, and with the atomic force microscopy of the skeletal network, the results of the present study provide the first evidence for the physiological function of calpain-1 in erythrocytes with therapeutic implications for calcium imbalance pathologies such as sickle cell disease.

Finite element analysis of wrinkling membranes

NASA Technical Reports Server (NTRS)

Miller, R. K.; Hedgepeth, J. M.; Weingarten, V. I.; Das, P.; Kahyai, S.

1984-01-01

The development of a nonlinear numerical algorithm for the analysis of stresses and displacements in partly wrinkled flat membranes, and its implementation on the SAP VII finite-element code are described. A comparison of numerical results with exact solutions of two benchmark problems reveals excellent agreement, with good convergence of the required iterative procedure. An exact solution of a problem involving axisymmetric deformations of a partly wrinkled shallow curved membrane is also reported.

Quantification of the fraction poorly deformable red blood cells using ektacytometry.

PubMed

Streekstra, G J; Dobbe, J G G; Hoekstra, A G

2010-06-21

We describe a method to obtain the fraction of poorly deformable red blood cells in a blood sample from the intensity pattern in an ektacytometer. In an ektacytometer red blood cells are transformed into ellipsoids by a shear flow between two transparent cylinders. The intensity pattern, due to a laser beam that is sent through the suspension, is projected on a screen. When measuring a healthy red blood cell population iso-intensity curves are ellipses with an axial ratio equal to that of the average red blood cell. In contrast poorly deformable cells result in circular iso-intensity curves. In this study we show that for mixtures of deformable and poorly deformable red blood cells, iso-intensity curves in the composite intensity pattern are neither elliptical nor circular but obtain cross-like shapes. We propose a method to obtain the fraction of poorly deformable red blood cells from those intensity patterns. Experiments with mixtures of poorly deformable and deformable red blood cells validate the method and demonstrate its accuracy. In a clinical setting our approach is potentially of great value for the detection of the fraction of sickle cells in blood samples of patients with sickle cell disease or to find a measure for the parasitemia in patients infected with malaria.

Mechanism of unassisted ion transport across membrane bilayers

NASA Technical Reports Server (NTRS)

Wilson, M. A.; Pohorille, A.

1996-01-01

To establish how charged species move from water to the nonpolar membrane interior and to determine the energetic and structural effects accompanying this process, we performed molecular dynamics simulations of the transport of Na+ and Cl- across a lipid bilayer located between two water lamellae. The total length of molecular dynamics trajectories generated for each ion was 10 ns. Our simulations demonstrate that permeation of ions into the membrane is accompanied by the formation of deep, asymmetric thinning defects in the bilayer, whereby polar lipid head groups and water penetrate the nonpolar membrane interior. Once the ion crosses the midplane of the bilayer the deformation "switches sides"; the initial defect slowly relaxes, and a defect forms in the outgoing side of the bilayer. As a result, the ion remains well solvated during the process; the total number of oxygen atoms from water and lipid head groups in the first solvation shell remains constant. A similar membrane deformation is formed when the ion is instantaneously inserted into the interior of the bilayer. The formation of defects considerably lowers the free energy barrier to transfer of the ion across the bilayer and, consequently, increases the permeabilities of the membrane to ions, compared to the rigid, planar structure, by approximately 14 orders of magnitude. Our results have implications for drug delivery using liposomes and peptide insertion into membranes.

Using NASTRAN to solve symmetric structures with nonsymmetric loads

NASA Technical Reports Server (NTRS)

Butler, T. G.

1982-01-01

A method for computation of reflective dihedral symmetry in symmetrical structures under nonsymmetric loads is described. The method makes it possible to confine the analysis to a half, a quarter, or an octagonal segment. The symmetry of elastic deformation is discussed, and antisymmetrical deformation is distinguished from nonsymmetrical deformation. Modes of deformation considered are axial, bending, membrane, and torsional deformation. Examples of one and two dimensional elements are presented and extended to three dimensional elements. The method of setting up a problem within NASTRAN is discussed. The technique is applied to a thick structure having quarter symmetry which was modeled with polyhedra and subjected to five distinct loads having varying degrees of symmetry.

AtMSL9 and AtMSL10: Sensors of plasma membrane tension in Arabidopsis roots.

PubMed

Peyronnet, Rémi; Haswell, Elizabeth S; Barbier-Brygoo, Hélène; Frachisse, Jean-Marie

2008-09-01

Plant cells, like those of animals and bacteria, are able to sense physical deformation of the plasma membrane. Mechanosensitive (MS) channels are proteins that transduce mechanical force into ion flux, providing a mechanism for the perception of mechanical stimuli such as sound, touch and osmotic pressure. We recently identified AtMSL9 and AtMSL10, two mechanosensitive channels in Arabidopsis thaliana, as molecular candidates for mechanosensing in higher plants.1 AtMSL9 and AtMSL10 are members of a family of proteins in Arabidopsis that are related to the bacterial MS channel MscS, termed MscS-Like (or MSL).2 MscS (Mechanosensitive channel of Small conductance) is one of the best-characterized MS channels, first identified as an electrophysiological activity in the plasma membrane (PM) of giant E. coli spheroplasts.3,4 Activation of MscS is voltage-independent, but responds directly to tension applied to the membrane and does not require other cellular proteins for this regulation.5,6 MscS family members are widely distributed throughout bacterial and archaeal genomes, are present in all plant genomes yet examined, and are found in selected fungal genomes.2,7,8 MscS homolgues have not yet been identified in animals.

A finite element method to compute three-dimensional equilibrium configurations of fluid membranes: Optimal parameterization, variational formulation and applications

NASA Astrophysics Data System (ADS)

Rangarajan, Ramsharan; Gao, Huajian

2015-09-01

We introduce a finite element method to compute equilibrium configurations of fluid membranes, identified as stationary points of a curvature-dependent bending energy functional under certain geometric constraints. The reparameterization symmetries in the problem pose a challenge in designing parametric finite element methods, and existing methods commonly resort to Lagrange multipliers or penalty parameters. In contrast, we exploit these symmetries by representing solution surfaces as normal offsets of given reference surfaces and entirely bypass the need for artificial constraints. We then resort to a Galerkin finite element method to compute discrete C1 approximations of the normal offset coordinate. The variational framework presented is suitable for computing deformations of three-dimensional membranes subject to a broad range of external interactions. We provide a systematic algorithm for computing large deformations, wherein solutions at subsequent load steps are identified as perturbations of previously computed ones. We discuss the numerical implementation of the method in detail and demonstrate its optimal convergence properties using examples. We discuss applications of the method to studying adhesive interactions of fluid membranes with rigid substrates and to investigate the influence of membrane tension in tether formation.

Special sensors for deformation measurements of different construction materials and structures

NASA Astrophysics Data System (ADS)

Glisic, Branko; Inaudi, Daniele; Kronenberg, Pascal; LLoret, Sandra; Vurpillot, Samuel

1999-05-01

SOFO is a fiber optic sensor system that allows the monitoring of micrometer deformations over measurement bases up to a few meters. It is particularly adapted to measure civil structures built with conventional civil engineering materials (concrete, steel and timber). It has been successfully tested in different types of structures such as bridges, tunnels and piles. The application of the system is however limited in some case when unusual materials are used in the construction and in other cases by the dimensions of standard SOFO sensors. To extend the domain of application of the current system, special sensors have been developed. In this paper we present four special SOFO sensors: long, membrane, thin and stiff sensors. The long sensor has a measurement basis of several tenths of meters and its purpose is the measurement of deformations in massive and large structures (dames, tunnels). The membrane sensor is for use on laminated materials (e.g. membrane roofing) and it is easy to install by simply gluing it to the structure to be monitored. Since standard sensors can not be used for thin mortar layers because of their cross- section, a thin sensor has been developed, too. Finally, the aim of the stiff sensor is to determine the hardening (solidification) time of concrete. This time is determined by comparing the deformations of a stiff and a standard sensor, closely placed in the concrete at the very early age. The design of these sensors is presented along with significant application examples.

Numerical study on 3D composite morphing actuators

NASA Astrophysics Data System (ADS)

Oishi, Kazuma; Saito, Makoto; Anandan, Nishita; Kadooka, Kevin; Taya, Minoru

2015-04-01

There are a number of actuators using the deformation of electroactive polymer (EAP), where fewer papers seem to have focused on the performance of 3D morphing actuators based on the analytical approach, due mainly to their complexity. The present paper introduces a numerical analysis approach on the large scale deformation and motion of a 3D half dome shaped actuator composed of thin soft membrane (passive material) and EAP strip actuators (EAP active coupon with electrodes on both surfaces), where the locations of the active EAP strips is a key parameter. Simulia/Abaqus Static and Implicit analysis code, whose main feature is the high precision contact analysis capability among structures, are used focusing on the whole process of the membrane to touch and wrap around the object. The unidirectional properties of the EAP coupon actuator are used as input data set for the material properties for the simulation and the verification of our numerical model, where the verification is made as compared to the existing 2D solution. The numerical results can demonstrate the whole deformation process of the membrane to wrap around not only smooth shaped objects like a sphere or an egg, but also irregularly shaped objects. A parametric study reveals the proper placement of the EAP coupon actuators, with the modification of the dome shape to induce the relevant large scale deformation. The numerical simulation for the 3D soft actuators shown in this paper could be applied to a wider range of soft 3D morphing actuators.

Optical Deformability as New Diagnostic Cell Marker

NASA Astrophysics Data System (ADS)

Guck, Jochen; Lincoln, Bryan; Schinkinger, Stefan; Wottawah, Falk; Moore, Samantha; Ananthakrishnan, Revathi; Kas, Josef

2002-03-01

The optical stretcher is a novel laser tool that can deform individual cells in rapid succession. When a cell is trapped between two counterpropagating laser beams the optically induced surface forces stretch the cell along the laser axis. The degree of stretching depends on the optical properties, which determine the forces, as well as the mechanical properties, which govern the response of the cell to the forces. Our results show that different cells can be distinguished based on their optical deformability, which naturally suggests using the optical deformability of cells as a novel cell marker. Many diseases are reflected in an altered cytoskeleton, which leads to a different optical deformability. An important example is the malignant transformation of cells, which is accompanied by a decrease in cytoskeletal integrity and, consequently, cell elasticity. Using optical deformability as cell marker holds the promise of earlier detection and improved diagnosis of cancer. In this context, the optical stretcher can be used as a diagnostic device to detect and sort abnormal cells. Future applications in the study of the normal differentiation of cells from stem cells to mature cells are envisioned.

Analytical and Experimental Characterization of Gravity Induced Deformations In Subscale Gossamer Structures

NASA Technical Reports Server (NTRS)

Johnston, John D.; Blandino, Joseph R.; McEvoy, Kiley C.

2004-01-01

The development of gossamer space structures such as solar sails and sunshields presents many challenges due to their large size and extreme flexibility. The post-deployment structural geometry exhibited during ground testing may significantly depart from the in-space configuration due to the presence of gravity-induced deformations (gravity sag) of lightly preloaded membranes. This paper describes a study carried out to characterize gravity sag in two subscale gossamer structures: a single quadrant from a 2 m, 4 quadrant square solar sail and a 1.7 m membrane layer from a multi-layer sunshield The behavior of the test articles was studied over a range of preloads and in several orientations with respect to gravity. An experimental study was carried out to measure the global surface profiles using photogrammetry, and nonlinear finite element analysis was used to predict the behavior of the test articles. Comparison of measured and predicted surface profiles shows that the finite dement analysis qualitatively predicts deformed shapes comparable to those observed in the laboratory. Quantitatively, finite element analysis predictions for peak gravity-induced deformations in both test articles were within 10% of measured values. Results from this study provide increased insight into gravity sag behavior in gossamer structures, and demonstrates the potential to analytically predict gravity-induced deformations to within reasonable accuracy.

The Antibacterial Activity of Coriolus versicolor Methanol Extract and Its Effect on Ultrastructural Changes of Staphylococcus aureus and Salmonella Enteritidis.

PubMed

Matijašević, Danka; Pantić, Milena; Rašković, Božidar; Pavlović, Vladimir; Duvnjak, Dunja; Sknepnek, Aleksandra; Nikšić, Miomir

2016-01-01

The antibacterial activity of methanol extract obtained from fruiting body of industrially grown basidiomycete Coriolus versicolor was examined. The Minimum Inhibitory Concentration (MIC) values against various bacteria ranged from 0.625 to 20 mg mL(-1). C. versicolor expressed bactericidal activity against both Gram-positive and Gram-negative bacteria. The growth curves of Staphylococcus aureus and Salmonella enterica serovar Enteritidis, measured at 630 nm, and confirmed with macrodilution method showed that the obtained extract could inhibit the growth of tested bacteria. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), and the loss of 260-nm-absorbing material were used to examine the ultrastructural changes in bacteria induced by the extract. When S. aureus was exposed to the MIC of C. versicolor, elongated and malformed cells were observed by SEM, while S. Enteritidis treated cells appeared shorter and aggregated with ruptured cell walls. TEM revealed the formation of non-membrane-enclosed bodies and depleted inner content of S. aureus. Larger and irregular periplasmic space and deformed and scattered components of the cell envelope were observed in treated S. Enteritidis. The loss of 260-nm-absorbing material indicated that the disruptive action of the extract on cytoplasmic membrane was more pronounced in S. aureus than in S. Enteritidis treated cells. The UV and FTIR spectrophotometric analyses revealed diverse composition of C. versicolor extract and high content of total phenolics. Altogether, mushroom extracts could be used to develop nutraceuticals or drugs effective against pathogenic microorganisms.

Alq3 coated silicon nanomembranes for cavity optomechanics

NASA Astrophysics Data System (ADS)

Fogliano, Francesco; Ortu, Antonio; Camposeo, Andrea; Pisignano, Dario; Ciampini, Donatella; Fuso, Francesco; Arimondo, E.

2016-09-01

The optomechanical properties of a silicon-nitride membrane mirror covered by Alq3 and Silver layers are investigated. Excitation at two laser wavelengths, 780 and 405 nm, corresponding to different absorptions of the multilayer, is examined. Such dual driving will lead to a more flexible optomechanical operation. Topographic reconstruction of the whole static membrane deformation and cooling of the membrane oscillations are reported. The cooling, observed for blue laser detuning and produced by bolometric forces, is deduced from the optomechanical damping of the membrane eigenfrequency. We determine the presence of different contributions to the photothermal response of the membrane.

Lipid Bilayer Vesicles with Numbers of Membrane-Linking Pores

NASA Astrophysics Data System (ADS)

Ken-ichirou Akashi,; Hidetake Miyata,

2010-06-01

We report that phospholipid membranes spontaneously formed in aqueous medium giant unilamellar vesicles (GUVs) possessing many membranous wormhole-like structures (membrane-linking pores, MLPs). By phase contract microscopy and confocal fluorescence microscopy, the structures of the MLPs, consisting of lipid bilayer, were resolvable, and a variety of vesicular shapes having many MLPs (a high genus topology) were found. These vesicles were stable but easily deformed by micromanipulation with a microneedle. We also observed the size reduction of the MLPs with the increase in membrane tension, which was qualitatively consistent with a prediction from a simple dynamical model.

Structural Stability of Light-harvesting Protein LH2 Adsorbed on Mesoporous Silica Supports.

PubMed

Shibuya, Yuuta; Itoh, Tetsuji; Matsuura, Shun-ichi; Yamaguchi, Akira

2015-01-01

In the present study, we examined the reversible thermal deformation of the membrane protein light-harvesting complex LH2 adsorbed on mesoporous silica (MPS) supports. The LH2 complex from Thermochromatium tepidum cells was conjugated to MPS supports with a series of pore diameter (2.4 to 10.6 nm), and absorption spectra of the resulting LH2/MPS conjugates were observed over a temperature range of 273 - 313 K in order to examine the structure of the LH2 adsorbed on the MPS support. The experimental results confirmed that a slight ellipsoidal deformation of LH2 was induced by adsorption on the MPS supports. On the other hand, the structural stability of LH2 was not perturbed by the adsorption. Since the pore diameter of MPS support did not influence the structural stability of LH2, it could be considered that the spatial confinement of LH2 in size-matches pore did not improve the structural stability of LH2.

Cell Deformation by Single-beam Acoustic Trapping: A Promising Tool for Measurements of Cell Mechanics

PubMed Central

Hwang, Jae Youn; Kim, Jihun; Park, Jin Man; Lee, Changyang; Jung, Hayong; Lee, Jungwoo; Shung, K. Kirk

2016-01-01

We demonstrate a noncontact single-beam acoustic trapping method for the quantification of the mechanical properties of a single suspended cell with label-free. Experimentally results show that the single-beam acoustic trapping force results in morphological deformation of a trapped cell. While a cancer cell was trapped in an acoustic beam focus, the morphological changes of the immobilized cell were monitored using bright-field imaging. The cell deformability was then compared with that of a trapped polystyrene microbead as a function of the applied acoustic pressure for a better understanding of the relationship between the pressure and degree of cell deformation. Cell deformation was found to become more pronounced as higher pressure levels were applied. Furthermore, to determine if this acoustic trapping method can be exploited in quantifying the cell mechanics in a suspension and in a non-contact manner, the deformability levels of breast cancer cells with different degrees of invasiveness due to acoustic trapping were compared. It was found that highly-invasive breast cancer cells exhibited greater deformability than weakly-invasive breast cancer cells. These results clearly demonstrate that the single-beam acoustic trapping technique is a promising tool for non-contact quantitative assessments of the mechanical properties of single cells in suspensions with label-free. PMID:27273365

Analysis of a bubble deformation process in a microcapsule by shock waves for developing DDS

NASA Astrophysics Data System (ADS)

Tamagawa, Masaaki; Morimoto, Kenshi

2012-09-01

This paper describes development of DDS (drug delivery systems) microcapsule using underwater shock waves, especially (1) making polymer microcapsules including a bubble and analysis of a bubble deformation process in a polymer capsule by pressure wave, (2) making liposome microcapsules with different elastic membrane and disintegration tests by ultrasonic waves.

The dielectric spectroscopy of human red blood cells: the differentiation of old from fresh cells.

PubMed

David, Marcelo; Levy, Evgeniya; Feldman, Yuri; Ben Ishai, Paul; Zelig, Orly; Yedgar, Saul; Barshtein, Gregory

2017-06-22

The objective of the study was to gauge the effect of storage lesions on the dielectric response of red blood cells (RBC), in particular those processes linked to deformations of the cellular membrane known as the β-dispersion. The dielectric response of RBC suspensions, exposed to blood-bank cold storage, was studied using time-domain dielectric spectroscopy (TDDS) in the frequency range of 500 kHz up to 1 GHz. The measured dielectric processes are characterized by their dielectric strength (Δε) and relaxation time (τ). Changes in the dielectric properties of the RBC suspensions due to storage-related lesions were evaluated. For a quantitative characterization of RBC lesions, we measured the deformability of fresh and stored RBC as expressed by their elongation ratio (ER), which was achieved under a shear stress of 3.0 Pa. The results show that the storage of RBC induced a statistically significant decrease of dielectric relaxation times. In addition, a sound correlation between the mean values of ER and the relaxation times was observed (Spearman's correlation coefficient ρ  =  0.847). We draw the conclusion that those alterations in the relaxation time are induced by changes in the shape of the RBC that happen during cold-storage. The evolution of the β-dispersion of RBC opens new possibilities in the blood bank inventory management.

Activation of the adenylyl cyclase/cyclic AMP/protein kinase A pathway in endothelial cells exposed to cyclic strain

NASA Technical Reports Server (NTRS)

Cohen, C. R.; Mills, I.; Du, W.; Kamal, K.; Sumpio, B. E.

1997-01-01

The aim of this study was to assess the involvement of the adenylyl cyclase/cyclic AMP/protein kinase A pathway (AC) in endothelial cells (EC) exposed to different levels of mechanical strain. Bovine aortic EC were seeded to confluence on flexible membrane-bottom wells. The membranes were deformed with either 150 mm Hg (average 10% strain) or 37.5 mm Hg (average 6% strain) vacuum at 60 cycles per minute (0.5 s strain; 0.5 s relaxation) for 0-60 min. The results demonstrate that at 10% average strain (but not 6% average strain) there was a 1.5- to 2.2-fold increase in AC, cAMP, and PKA activity by 15 min when compared to unstretched controls. Further studies revealed an increase in cAMP response element binding protein in EC subjected to the 10% average strain (but not 6% average strain). These data support the hypothesis that cyclic strain activates the AC/cAMP/PKA signal transduction pathway in EC which may occur by exceeding a strain threshold and suggest that cyclic strain may stimulate the expression of genes containing cAMP-responsive promoter elements.

Nanoscale diffractive probing of strain dynamics in ultrafast transmission electron microscopy

PubMed Central

Feist, Armin; Rubiano da Silva, Nara; Liang, Wenxi; Ropers, Claus; Schäfer, Sascha

2018-01-01

The control of optically driven high-frequency strain waves in nanostructured systems is an essential ingredient for the further development of nanophononics. However, broadly applicable experimental means to quantitatively map such structural distortion on their intrinsic ultrafast time and nanometer length scales are still lacking. Here, we introduce ultrafast convergent beam electron diffraction with a nanoscale probe beam for the quantitative retrieval of the time-dependent local deformation gradient tensor. We demonstrate its capabilities by investigating the ultrafast acoustic deformations close to the edge of a single-crystalline graphite membrane. Tracking the structural distortion with a 28-nm/700-fs spatio-temporal resolution, we observe an acoustic membrane breathing mode with spatially modulated amplitude, governed by the optical near field structure at the membrane edge. Furthermore, an in-plane polarized acoustic shock wave is launched at the membrane edge, which triggers secondary acoustic shear waves with a pronounced spatio-temporal dependency. The experimental findings are compared to numerical acoustic wave simulations in the continuous medium limit, highlighting the importance of microscopic dissipation mechanisms and ballistic transport channels. PMID:29464187

Length and sequence dependence in the association of Huntingtin protein with lipid membranes

NASA Astrophysics Data System (ADS)

Jawahery, Sudi; Nagarajan, Anu; Matysiak, Silvina

2013-03-01

There is a fundamental gap in our understanding of how aggregates of mutant Huntingtin protein (htt) with overextended polyglutamine (polyQ) sequences gain the toxic properties that cause Huntington's disease (HD). Experimental studies have shown that the most important step associated with toxicity is the binding of mutant htt aggregates to lipid membranes. Studies have also shown that flanking amino acid sequences around the polyQ sequence directly affect interactions with the lipid bilayer, and that polyQ sequences of greater than 35 glutamine repeats in htt are a characteristic of HD. The key steps that determine how flanking sequences and polyQ length affect the structure of lipid bilayers remain unknown. In this study, we use atomistic molecular dynamics simulations to study the interactions between lipid membranes of varying compositions and polyQ peptides of varying lengths and flanking sequences. We find that overextended polyQ interactions do cause deformation in model membranes, and that the flanking sequences do play a role in intensifying this deformation by altering the shape of the affected regions.

Nanoscale diffractive probing of strain dynamics in ultrafast transmission electron microscopy.

PubMed

Feist, Armin; Rubiano da Silva, Nara; Liang, Wenxi; Ropers, Claus; Schäfer, Sascha

2018-01-01

The control of optically driven high-frequency strain waves in nanostructured systems is an essential ingredient for the further development of nanophononics. However, broadly applicable experimental means to quantitatively map such structural distortion on their intrinsic ultrafast time and nanometer length scales are still lacking. Here, we introduce ultrafast convergent beam electron diffraction with a nanoscale probe beam for the quantitative retrieval of the time-dependent local deformation gradient tensor. We demonstrate its capabilities by investigating the ultrafast acoustic deformations close to the edge of a single-crystalline graphite membrane. Tracking the structural distortion with a 28-nm/700-fs spatio-temporal resolution, we observe an acoustic membrane breathing mode with spatially modulated amplitude, governed by the optical near field structure at the membrane edge. Furthermore, an in-plane polarized acoustic shock wave is launched at the membrane edge, which triggers secondary acoustic shear waves with a pronounced spatio-temporal dependency. The experimental findings are compared to numerical acoustic wave simulations in the continuous medium limit, highlighting the importance of microscopic dissipation mechanisms and ballistic transport channels.

Fiber-reinforced dielectric elastomer laminates with integrated function of actuating and sensing

NASA Astrophysics Data System (ADS)

Li, Tiefeng; Xie, Yuhan; Li, Chi; Yang, Xuxu; Jin, Yongbin; Liu, Junjie; Huang, Xiaoqiang

2015-04-01

The natural limbs of animals and insects integrate muscles, skins and neurons, providing both the actuating and sensing functions simultaneously. Inspired by the natural structure, we present a novel structure with integrated function of actuating and sensing with dielectric elastomer (DE) laminates. The structure can deform when subjected to high voltage loading and generate corresponding output signal in return. We investigate the basic physical phenomenon of dielectric elastomer experimentally. It is noted that when applying high voltage, the actuating dielectric elastomer membrane deforms and the sensing dielectric elastomer membrane changes the capacitance in return. Based on the concept, finite element method (FEM) simulation has been conducted to further investigate the electromechanical behavior of the structure.

Analysis of the variation in the determination of the shear modulus of the erythrocyte membrane: Effects of the constitutive law and membrane modeling

PubMed Central

Dimitrakopoulos, P.

2013-01-01

Despite research spanning several decades, the exact value of the shear modulus Gs of the erythrocyte membrane is still ambiguous, and a wealth of studies, using measurements based on micropipette aspirations, ektacytometry systems and other flow chambers, and optical tweezers as well as application of several models have found different average values in the range 2–10 µN/m. Our study shows that different methodologies have predicted the correct shear modulus for the specific membrane modeling employed, i.e. the variation in the shear modulus determination results from the specific membrane modeling. Available experimental findings from ektacytometry systems and optical tweezers suggest that the dynamics of the erythrocyte membrane is strain-hardening at both moderate and large deformations. Thus the erythrocyte shear modulus cannot be determined accurately using strain-softening models (such as the neo-Hookean and Evans laws) or strain-softening/strain-hardening models (such as the Yeoh law) which overestimate the erythrocyte shear modulus. According to our analysis, the only available strain-hardening constitutive law, the Skalak et al. law, is able to match well both deformation-shear rate data from ektacytometry and force-extension data from optical tweezers at moderate and large strains, using an average value of the shear modulus of Gs = 2.4–2.75 µN/m, i.e. very close to that found in the linear regime of deformations via force-extension data from optical tweezers, Gs = 2.5±0.4 µN/m. In addition, our analysis suggests that a standard deviation in Gs of 0.4–0.5 µN/m (owing to the inherent differences between erythrocytes within a large population) describes well the findings from optical tweezers at small and large strains as well as from micro-pipette aspirations. PMID:22680508

IB-LBM simulation of the haemocyte dynamics in a stenotic capillary.

PubMed

Yuan-Qing, Xu; Xiao-Ying, Tang; Fang-Bao, Tian; Yu-Hua, Peng; Yong, Xu; Yan-Jun, Zeng

2014-01-01

To study the behaviour of a haemocyte when crossing a stenotic capillary, the immersed boundary-lattice Boltzmann method was used to establish a quantitative analysis model. The haemocyte was assumed to be spherical and to have an elastic cell membrane, which can be driven by blood flow to adopt a highly deformable character. In the stenotic capillary, the spherical blood cell was stressed both by the flow and the wall dimension, and the cell shape was forced to be stretched to cross the stenosis. Our simulation investigated the haemocyte crossing process in detail. The velocity and pressure were anatomised to obtain information on how blood flows through a capillary and to estimate the degree of cell damage caused by excessive pressure. Quantitative velocity analysis results demonstrated that a large haemocyte crossing a small stenosis would have a noticeable effect on blood flow, while quantitative pressure distribution analysis results indicated that the crossing process would produce a special pressure distribution in the cell interior and to some extent a sudden change between the cell interior and the surrounding plasma.

Mechanically activated artificial cell by using microfluidics

NASA Astrophysics Data System (ADS)

Ho, Kenneth K. Y.; Lee, Lap Man; Liu, Allen P.

2016-09-01

All living organisms sense mechanical forces. Engineering mechanosensitive artificial cell through bottom-up in vitro reconstitution offers a way to understand how mixtures of macromolecules assemble and organize into a complex system that responds to forces. We use stable double emulsion droplets (aqueous/oil/aqueous) to prototype mechanosensitive artificial cells. In order to demonstrate mechanosensation in artificial cells, we develop a novel microfluidic device that is capable of trapping double emulsions into designated chambers, followed by compression and aspiration in a parallel manner. The microfluidic device is fabricated using multilayer soft lithography technology, and consists of a control layer and a deformable flow channel. Deflections of the PDMS membrane above the main microfluidic flow channels and trapping chamber array are independently regulated pneumatically by two sets of integrated microfluidic valves. We successfully compress and aspirate the double emulsions, which result in transient increase and permanent decrease in oil thickness, respectively. Finally, we demonstrate the influx of calcium ions as a response of our mechanically activated artificial cell through thinning of oil. The development of a microfluidic device to mechanically activate artificial cells creates new opportunities in force-activated synthetic biology.

Cellular interactions with tissue-engineered microenvironments and nanoparticles

NASA Astrophysics Data System (ADS)

Pan, Zhi

Tissue-engineered hydrogels composed of intermolecularlly crosslinked hyaluronan (HA-DTPH) and fibronectin functional domains (FNfds) were applied as a physiological relevant ECM mimic with controlled mechanical and biochemical properties. Cellular interactions with this tissue-engineered environment, especially physical interactions (cellular traction forces), were quantitatively measured by using the digital image speckle correlation (DISC) technique and finite element method (FEM). By correlating with other cell functions such as cell morphology and migration, a comprehensive structure-function relationship between cells and their environments was identified. Furthermore, spatiotemporal redistribution of cellular traction stresses was time-lapse measured during cell migration to better understand the dynamics of cell mobility. The results suggest that the reinforcement of the traction stresses around the nucleus, as well as the relaxation of nuclear deformation, are critical steps during cell migration, serving as a speed regulator, which must be considered in any dynamic molecular reconstruction model of tissue cell migration. Besides single cell migration, en masse cell migration was studied by using agarose droplet migration assay. Cell density was demonstrated to be another important parameter to influence cell behaviors besides substrate properties. Findings from these studies will provide fundamental design criteria to develop novel and effective tissue-engineered constructs. Cellular interactions with rutile and anatase TiO2 nanoparticles were also studied. These particles can penetrate easily through the cell membrane and impair cell function, with the latter being more damaging. The exposure to nanoparticles was found to decrease cell area, cell proliferation, motility, and contractility. To prevent this, a dense grafted polymer brush coating was applied onto the nanoparticle surface. These modified nanoparticles failed to adhere to and penetrate through the cell membrane. As a consequence, the coating effectively decreased reactive oxygen species (ROS) formation and protected the cells. Considering the broad applications of these nanoparticles in personal health care products, the functionalized polymer coating will likely play an important role in protecting cells and tissue from damage.

Steel reinforced composite silicone membranes and its integration to microfluidic oxygenators for high performance gas exchange.

PubMed

Matharoo, Harpreet; Dabaghi, Mohammadhossein; Rochow, Niels; Fusch, Gerhard; Saraei, Neda; Tauhiduzzaman, Mohammed; Veldhuis, Stephen; Brash, John; Fusch, Christoph; Selvaganapathy, P Ravi

2018-01-01

Respiratory distress syndrome (RDS) is one of the main causes of fatality in newborn infants, particularly in neonates with low birth-weight. Commercial extracorporeal oxygenators have been used for low-birth-weight neonates in neonatal intensive care units. However, these oxygenators require high blood volumes to prime. In the last decade, microfluidics oxygenators using enriched oxygen have been developed for this purpose. Some of these oxygenators use thin polydimethylsiloxane (PDMS) membranes to facilitate gas exchange between the blood flowing in the microchannels and the ambient air outside. However, PDMS is elastic and the thin membranes exhibit significant deformation and delamination under pressure which alters the architecture of the devices causing poor oxygenation or device failure. Therefore, an alternate membrane with high stability, low deformation under pressure, and high gas exchange was desired. In this paper, we present a novel composite membrane consisting of an ultra-thin stainless-steel mesh embedded in PDMS, designed specifically for a microfluidic single oxygenator unit (SOU). In comparison to homogeneous PDMS membranes, this composite membrane demonstrated high stability, low deformation under pressure, and high gas exchange. In addition, a new design for oxygenator with sloping profile and tapered inlet configuration has been introduced to achieve the same gas exchange at lower pressure drops. SOUs were tested by bovine blood to evaluate gas exchange properties. Among all tested SOUs, the flat design SOU with composite membrane has the highest oxygen exchange of 40.32 ml/min m 2 . The superior performance of the new device with composite membrane was demonstrated by constructing a lung assist device (LAD) with a low priming volume of 10 ml. The LAD was achieved by the oxygen uptake of 0.48-0.90 ml/min and the CO 2 release of 1.05-2.27 ml/min at blood flow rates ranging between 8 and 48 ml/min. This LAD was shown to increase the oxygen saturation level by 25% at the low pressure drop of 29 mm Hg. Finally, a piglet was used to test the gas exchange capacity of the LAD in vivo . The animal experiment results were in accordance with in-vitro results, which shows that the LAD is capable of providing sufficient gas exchange at a blood flow rate of ∼24 ml/min.

Analysing intracellular deformation of polymer capsules using structured illumination microscopy

NASA Astrophysics Data System (ADS)

Chen, Xi; Cui, Jiwei; Sun, Huanli; Müllner, Markus; Yan, Yan; Noi, Ka Fung; Ping, Yuan; Caruso, Frank

2016-06-01

Understanding the behaviour of therapeutic carriers is important in elucidating their mechanism of action and how they are processed inside cells. Herein we examine the intracellular deformation of layer-by-layer assembled polymer capsules using super-resolution structured illumination microscopy (SIM). Spherical- and cylindrical-shaped capsules were studied in three different cell lines, namely HeLa (human epithelial cell line), RAW264.7 (mouse macrophage cell line) and differentiated THP-1 (human monocyte-derived macrophage cell line). We observed that the deformation of capsules was dependent on cell line, but independent of capsule shape. This suggests that the mechanical forces, which induce capsule deformation during cell uptake, vary between cell lines, indicating that the capsules are exposed to higher mechanical forces in HeLa cells, followed by RAW264.7 and then differentiated THP-1 cells. Our study demonstrates the use of super-resolution SIM in analysing intracellular capsule deformation, offering important insights into the cellular processing of drug carriers in cells and providing fundamental knowledge of intracellular mechanobiology. Furthermore, this study may aid in the design of novel drug carriers that are sensitive to deformation for enhanced drug release properties.Understanding the behaviour of therapeutic carriers is important in elucidating their mechanism of action and how they are processed inside cells. Herein we examine the intracellular deformation of layer-by-layer assembled polymer capsules using super-resolution structured illumination microscopy (SIM). Spherical- and cylindrical-shaped capsules were studied in three different cell lines, namely HeLa (human epithelial cell line), RAW264.7 (mouse macrophage cell line) and differentiated THP-1 (human monocyte-derived macrophage cell line). We observed that the deformation of capsules was dependent on cell line, but independent of capsule shape. This suggests that the mechanical forces, which induce capsule deformation during cell uptake, vary between cell lines, indicating that the capsules are exposed to higher mechanical forces in HeLa cells, followed by RAW264.7 and then differentiated THP-1 cells. Our study demonstrates the use of super-resolution SIM in analysing intracellular capsule deformation, offering important insights into the cellular processing of drug carriers in cells and providing fundamental knowledge of intracellular mechanobiology. Furthermore, this study may aid in the design of novel drug carriers that are sensitive to deformation for enhanced drug release properties. Electronic supplementary information (ESI) available: Additional figures. See DOI: 10.1039/c6nr02151d

A microfluidic distribution system for an array of hollow microneedles

NASA Astrophysics Data System (ADS)

Hoel, Antonin; Baron, Nolwenn; Cabodevila, Gonzalo; Jullien, Marie-Caroline

2008-06-01

We report a microfluidic device able to control the ejection of fluid through a matrix of out-of-plane microneedles. The device comprises a matrix of open dispensing units connected to needles and filled by a common filling system. A deformable membrane (e.g. in PDMS) is brought into contact with the dispensing units. Pressure exerted on the deformable membrane closes (and thus individualizes) each dispensing unit and provokes the ejection of the dispensing unit content through the outlets. Sufficient pressure over the deformable membrane ensures that all dispensing units deliver a fixed volume (their content) irrespective of the hydrodynamic pressure outside the dispensing unit outlet. The size of the ensemble matrix of dispensing units, the number of liquid reservoirs, as well as the material can vary depending on the considered application of the device or on the conditions of use. In the present paper, the liquid reservoirs are geometrically identical. The geometrical parameters of the device are optimized to avoid as much dead volume as possible, as it was to handle plasmid DNA solutions which are very expensive. The conception, the fabrication and the experimental results are described in this paper. Our prototype is conceived to inject in a uniform way 10 µl of drug through 100 microneedles distributed over 1 cm2.

Gap junctions are selectively associated with interlocking ball-and-sockets but not protrusions in the lens.

PubMed

Biswas, Sondip K; Lee, Jai Eun; Brako, Lawrence; Jiang, Jean X; Lo, Woo-Kuen

2010-11-09

Ball-and-sockets and protrusions are specialized interlocking membrane domains between lens fibers of all species studied. Ball-and-sockets and protrusions are similar in their shape, size, and surface morphology, and are traditionally believed to play a key role in maintaining fiber-to-fiber stability. Here, we evaluate the hypothesis that ball-and-sockets and protrusions possess important structural and functional differences during fiber cell differentiation and maturation. Intact lenses of leghorn chickens (E7 days to P62 weeks old) and rhesus monkeys (1.5-20 years old) were studied with SEM, freeze-fracture TEM, freeze-fracture immunogold labeling (FRIL), and filipin cytochemistry for membrane cholesterol detection. SEM showed that ball-and-sockets were distributed along the long and short sides of hexagonal fiber cells, whereas protrusions were located along the cell corners, from superficial to deep cortical regions in both chicken and monkey lenses. Importantly, by freeze-fracture TEM, we discovered the selective association of gap junctions with all ball-and-sockets examined, but not with protrusions, in both species. In the embryonic chicken lens (E18), the abundant distribution of ball-and-socket gap junctions was regularly found in an approximate zone extending at least 300 μm deep from the equatorial surface of the superficial cortical fibers. Many ball-and-socket gap junctions often protruded deeply into neighboring cells. However, in the mature fibers of monkey lenses, several ball-and-sockets exhibited only partial occupancy of gap junctions with disorganized connexons, possibly due to degradation of gap junctions during fiber maturation and aging. FRIL analysis confirmed that both connexin46 (Cx46) and connexin50 (Cx50) antibodies specifically labeled ball-and-socket gap junctions, but not protrusions. Furthermore, filipin cytochemistry revealed that the ball-and-socket gap junctions contained different amounts of cholesterol (i.e., cholesterol-rich versus cholesterol-free) as seen with the filipin-cholesterol-complexes (FCC) in different cortical regions during maturation. In contrast, the protrusions contained consistently high cholesterol amounts (i.e., 402 FCCs/μm2 membrane) which were approximately two times greater than that of the cholesterol-rich gap junctions (i.e., 188 FCCs/μm2 membrane) found in ball-and-sockets. Gap junctions are regularly associated with all ball-and-sockets examined in metabolically active young cortical fibers, but not with protrusions, in both chicken and monkey lenses. Since these unique gap junctions often protrude deeply into neighboring cells to increase membrane surface areas, they may significantly facilitate cell-to-cell communication between young cortical fiber cells. In particular, the large number of ball-and-socket gap junctions found near the equatorial region may effectively facilitate the flow of outward current toward the equatorial surface for internal circulation of ions in the lens. In contrast, a consistent distribution of high concentrations of cholesterol in protrusions would make the protrusion membrane less deformable and would be more suitable for maintaining fiber-to-fiber stability during visual accommodation. Thus, the ball-and-sockets and protrusions are two structurally and functionally distinct membrane domains in the lens.

Gap junctions are selectively associated with interlocking ball-and-sockets but not protrusions in the lens

PubMed Central

Biswas, Sondip K.; Lee, Jai Eun; Brako, Lawrence; Jiang, Jean X.

2010-01-01

Purpose Ball-and-sockets and protrusions are specialized interlocking membrane domains between lens fibers of all species studied. Ball-and-sockets and protrusions are similar in their shape, size, and surface morphology, and are traditionally believed to play a key role in maintaining fiber-to-fiber stability. Here, we evaluate the hypothesis that ball-and-sockets and protrusions possess important structural and functional differences during fiber cell differentiation and maturation. Methods Intact lenses of leghorn chickens (E7 days to P62 weeks old) and rhesus monkeys (1.5–20 years old) were studied with SEM, freeze-fracture TEM, freeze-fracture immunogold labeling (FRIL), and filipin cytochemistry for membrane cholesterol detection. Results SEM showed that ball-and-sockets were distributed along the long and short sides of hexagonal fiber cells, whereas protrusions were located along the cell corners, from superficial to deep cortical regions in both chicken and monkey lenses. Importantly, by freeze-fracture TEM, we discovered the selective association of gap junctions with all ball-and-sockets examined, but not with protrusions, in both species. In the embryonic chicken lens (E18), the abundant distribution of ball-and-socket gap junctions was regularly found in an approximate zone extending at least 300 μm deep from the equatorial surface of the superficial cortical fibers. Many ball-and-socket gap junctions often protruded deeply into neighboring cells. However, in the mature fibers of monkey lenses, several ball-and-sockets exhibited only partial occupancy of gap junctions with disorganized connexons, possibly due to degradation of gap junctions during fiber maturation and aging. FRIL analysis confirmed that both connexin46 (Cx46) and connexin50 (Cx50) antibodies specifically labeled ball-and-socket gap junctions, but not protrusions. Furthermore, filipin cytochemistry revealed that the ball-and-socket gap junctions contained different amounts of cholesterol (i.e., cholesterol-rich versus cholesterol-free) as seen with the filipin-cholesterol-complexes (FCC) in different cortical regions during maturation. In contrast, the protrusions contained consistently high cholesterol amounts (i.e., 402 FCCs/μm2 membrane) which were approximately two times greater than that of the cholesterol-rich gap junctions (i.e., 188 FCCs/μm2 membrane) found in ball-and-sockets. Conclusions Gap junctions are regularly associated with all ball-and-sockets examined in metabolically active young cortical fibers, but not with protrusions, in both chicken and monkey lenses. Since these unique gap junctions often protrude deeply into neighboring cells to increase membrane surface areas, they may significantly facilitate cell-to-cell communication between young cortical fiber cells. In particular, the large number of ball-and-socket gap junctions found near the equatorial region may effectively facilitate the flow of outward current toward the equatorial surface for internal circulation of ions in the lens. In contrast, a consistent distribution of high concentrations of cholesterol in protrusions would make the protrusion membrane less deformable and would be more suitable for maintaining fiber-to-fiber stability during visual accommodation. Thus, the ball-and-sockets and protrusions are two structurally and functionally distinct membrane domains in the lens. PMID:21139982

[Peritubular capillary injury in Chinese herb guan-mu-tong-induced acute tubular necrosis].

PubMed

Yang, Li; Li, Xiao-mei; Wang, Su-xia; Wang, Hai-yan

2005-07-01

To explore the role and mechanisms of peritubular capillary (PTC) injury in the progression of Chinese Herb guan-mu-tong (GMT, aristolochiae manshuriensis kom) induced acute tubular necrosis (GMT-ATN). Renal biopsy tissue from 4 cases of GMT-ATN and 5 cases of antibiotic induced ATN (A-ATN) were included in the study. Tubulointerstitial injury was semi-quantitatively assessed. Immunohistochemical SP method was applied to reveal PTC as well as the expression of vascular endothelial growth factor (VEGF). Ultra microstructure of endothelial cells and basement membrane of PTC was detected by electronic microscopy (EM). 5 cases of minor mesangioproliferative non-IgA glomerulonephritis were selected as a control group. The density of PTC was decreased significantly in GMT-ATN, as compared with the A-ATN and control group (211.08 +/- 56.15 vs 413.54 +/- 66.59, 536.62 +/- 68.38, P < 0.01). Dilated and deformed PTC lumina were noted in GMT-ATN with some endothelial cells and basement membrane partially disrupted. Most endothelial cells were found to be swollen with vacuoles dispersed in the cell plasma. The basement membrane was partially shrunk and thickened. The expression of VEGF in renal tubular epithelial cells (RTEC) was much less in the GMT-ATN than that in A-ATN group 2.1 (0-3.86)% vs [42.5 (31.33-60.25)%, P < 0.01], even though it was higher than that in the control group [23.1 (18.2-39.5)%, P < 0.01]; the expression was correlated with PTC density. Close correlation was also found between RTEC regeneration and PTC density, as well as VEGF expression (r = 0.880 and 0.802 respectively, P < 0.01). PTC was markedly injured in GMT-ATN; this could be one of the cause for the continuously progressing tubulointerstitial damage. The low expression of VEGF in RTEC might contribute to the PTC injury process.

Numerical simulation of the pairwise interaction of deformable cells during migration in a microchannel

NASA Astrophysics Data System (ADS)

Lan, Hongzhi; Khismatullin, Damir B.

2014-07-01

Leukocytes and other circulating cells deform and move relatively to the channel flow in the lateral and translational directions. Their migratory property is important in immune response, hemostasis, cancer progression, delivery of nutrients, and microfluidic technologies such as cell separation and enrichment, and flow cytometry. Using our three-dimensional computational algorithm for multiphase viscoelastic flow, we have investigated the effect of pairwise interaction on the lateral and translational migration of circulating cells in a microchannel. The numerical simulation data show that when two cells with the same size and small separation distance interact, repulsive interaction take place until they reach the same lateral equilibrium position. During this process, they undergo swapping or passing, depending on the initial separation distance between each other. The threshold value of this distance increases with cell deformation, indicating that the cells experiencing larger deformation are more likely to swap. When a series of closely spaced cells with the same size are considered, they generally undergo damped oscillation in both lateral and translational directions until they reach equilibrium positions where they become evenly distributed in the flow direction (self-assembly phenomenon). A series of cells with a large lateral separation distance could collide repeatedly with each other, eventually crossing the centerline and entering the other side of the channel. For a series of cells with different deformability, more deformable cells, upon impact with less deformable cells, move to an equilibrium position closer to the centerline. The results of our study show that the bulk deformation of circulating cells plays a key role in their migration in a microchannel.

Increased erythrocyte deformability in fetal erythropoiesis and in erythrocytes deficient in glucose-6-phosphate dehydrogenase and other glycolytic enzymes.

PubMed

Johnson, R M; Panchoosingh, H; Goyette, G; Ravindranath, Y

1999-01-01

Erythrocyte deformability was determined in more than 500 clinical samples, and was found to be elevated in conditions in which fetal-like red cells are produced: aplastic anemia (3/3 cases), myelodysplastic syndromes, polycythemias, sickle cell anemia during treatment with hydroxyurea, paroxysmal nocturnal hemoglobinuria, and recovery from B12 deficiency. Elevated deformability was observed in neonatal erythrocytes, and during recovery from transient erythroblastopenia of childhood, when fetal-like red cells are known to be produced. Increased deformability appears to be a feature of fetal and fetal-like red cells. Forty-eight cases of enzymatically verified glucose-6-phosphate (G-6-PD) deficiency were also examined. Thirty out of 32 G-6-PD(A-) individuals, including both heterozygotes and hemizygotes, exhibited increased deformability during the steady state. In contrast, G-6-PD(Med) hemizygotes had normal deformability. Increased deformability was also found in G-6-PD(Huron) (n=3), G-6-PD(Wayne) (n=4), triose phosphate isomerase deficiency (n=2), and pyruvate kinase deficiency (n=2). An elevated osmoscan was found in more than 90% of female G-6-PD heterozygotes, affording a simple screening test for heterozygotes. Deformability remained high during hemolytic episodes, when older enzyme deficient cells are removed from the circulation. In four cases of G-6-PD deficiency with normal deformability, evidence for co-existing hereditary spherocytosis was found. The combination of conditions with opposing effects on deformability resulted in nearly normal deformability. Because increased red cell deformability is a feature of fetal erythrocytes, these results suggest that the red cells in many cases of glycolytic enzyme deficiency are fetal-like.

Possible Role of Non-Muscle Alpha-Actinins in Muscle Cell Mechanosensitivity

PubMed Central

Ogneva, Irina V.; Biryukov, Nikolay S.; Leinsoo, Toomas A.; Larina, Irina M.

2014-01-01

The main hypothesis suggested that changes in the external mechanical load would lead to different deformations of the submembranous cytoskeleton and, as a result, dissociation of different proteins from its structure (induced by increased/decreased mechanical stress). The study subjects were fibers of the soleus muscle and cardiomyocytes of Wistar rats. Changes in external mechanical conditions were reconstructed by means of antiorthostatic suspension of the animals by their tails for 6, 12, 18, 24 and 72 hours. Transversal stiffness was measured by atomic force microscopy imaging; beta-, gamma-actin, alpha-actinin 1 and alpha-actinin 4 levels in membranous and cytoplasmic fractions were quantified by Western blot analysis; expression rates of the corresponding genes were studied using RT-PCR. Results: In 6 hours, alpha-actinin 1 and alpha-actinin 4 levels decreased in the membranous fraction of proteins of cardiomyocytes and soleus muscle fibers, respectively, but increased in the cytoplasmic fraction of the abovementioned cells. After 6–12 hours of suspension, the expression rates of beta-, gamma-actin, alpha-actinin 1 and alpha-actinin 4 were elevated in the soleus muscle fibers, but the alpha-actinin 1 expression rate returned to the reference level in 72 hours. After 18–24 hours, the expression rates of beta-actin and alpha-actinin 4 increased in cardiomyocytes, while the alpha-actinin 1 expression rate decreased in soleus muscle fibers. After 12 hours, the beta- and gamma-actin content dropped in the membranous fraction and increased in the cytoplasmic protein fractions from both cardiomyocytes and soleus muscle fibers. The stiffness of both cell types decreased after the same period of time. Further, during the unloading period the concentration of nonmuscle actin and different isoforms of alpha-actinins increased in the membranous fraction from cardiomyocytes. At the same time, the concentration of the abovementioned proteins decreased in the soleus muscle fibers. PMID:24780915

How deep cells feel: Mean-field Computations and Experiments

NASA Astrophysics Data System (ADS)

Buxboim, Amnon; Sen, Shamik; Discher, Dennis E.

2009-03-01

Most cells in solid tissues exert contractile forces that mechanically couple them to elastic surroundings and that significantly influence cell adhesion, cytoskeletal organization and differentiation. However, strains within the depths of matrices are often unclear and are likely relevant to thin matrices, such as basement membranes, relative to cell size as well as to defining how far cells can ``feel.'' We present experimental results for cell spreading on thin, ligand- coated gels and for prestress in stem cells in relation to gel stiffness. Matrix thickness affects cell spread area, focal adhesions and cytoskeleton organization in stem cells, which we will compare to differentiated cells. We introduce a finite element computation to estimate the elastostatic deformations within the matrix on which a cell is placed. Interfacial strains between cell and matrix show large deviations only when soft matrices are a fraction of cell dimensions, proving consistent with experiments. 3-D cell morphologies that model stem cell-derived neurons, myoblasts, and osteoblasts show that a cylinder-shaped myoblast induces the highest strains, consistent with the prominent contractility of muscle. Groups of such cells show a weak crosstalk via matrix strains only when cells are much closer than a cell-width. Cells thus feel on length scales closer to that of adhesions than on cellular scales.

Equilibrium of fluid membranes endowed with orientational order

NASA Astrophysics Data System (ADS)

Kumar Alageshan, Jaya; Chakrabarti, Buddhapriya; Hatwalne, Yashodhan

2017-04-01

Minimization of the low-temperature elastic free-energy functional of orientationlly ordered membranes involves independent variation of the membrane-shape, while keeping the orientational order on it (its texture) fixed. We propose an operational, coordinate-independent method for implementing such a variation. Using the Nelson-Peliti formulation of elasticity that emphasizes the interplay between geometry, topology, and thermal fluctuations of orientationally ordered membranes, we minimize the elastic free energy to obtain equations governing their equilibrium shape, together with associated free boundary conditions. Our results are essential for understanding and predicting equilibrium shapes as well as textures of membranes and vesicles; particularly under conditions in which shape deformations are large.

Robust Wave-front Correction in a Small Scale Adaptive Optics System Using a Membrane Deformable Mirror

NASA Astrophysics Data System (ADS)

Choi, Y.; Park, S.; Baik, S.; Jung, J.; Lee, S.; Yoo, J.

A small scale laboratory adaptive optics system using a Shack-Hartmann wave-front sensor (WFS) and a membrane deformable mirror (DM) has been built for robust image acquisition. In this study, an adaptive limited control technique is adopted to maintain the long-term correction stability of an adaptive optics system. To prevent the waste of dynamic correction range for correcting small residual wave-front distortions which are inefficient to correct, the built system tries to limit wave-front correction when a similar small difference wave-front pattern is repeatedly generated. Also, the effect of mechanical distortion in an adaptive optics system is studied and a pre-recognition method for the distortion is devised to prevent low-performance system operation. A confirmation process for a balanced work assignment among deformable mirror (DM) actuators is adopted for the pre-recognition. The corrected experimental results obtained by using a built small scale adaptive optics system are described in this paper.

α-Helix Unwinding as Force Buffer in Spectrins.

PubMed

Takahashi, Hirohide; Rico, Felix; Chipot, Christophe; Scheuring, Simon

2018-03-27

Spectrins are cytoskeletal proteins located at the inner face of the plasma membrane, making connections between membrane anchors and the actin cortex, and between actin filaments. Spectrins share a common structure forming a bundle of 3 α-helices and play a major role during cell deformation. Here, we used high-speed force spectroscopy and steered molecular dynamics simulations to understand the mechanical stability of spectrin, revealing a molecular force buffering function. We find that spectrin acts as a soft spring at short extensions (70-100 Å). Under continuous external stretching, its α-helices unwind, leading to a viscous mechanical response over larger extensions (100-300 Å), represented by a constant-force plateau in force/extension curves. This viscous force buffering emerges from a quasi-equilibrium competition between disruption and re-formation of α-helical hydrogen bonds. Our results suggest that, in contrast to β-sheet proteins, which unfold in a catastrophic event, α-helical spectrins dominantly unwind, providing a viscous force buffer over extensions about 5 times their folded length.

Photodynamic damage to cartilage and synovial tissue grafted on a chick's chorioallantoic membrane

NASA Astrophysics Data System (ADS)

Fisher, M.; Nahir, A. M.; Kimel, Sol

1997-09-01

Rheumatoid arthritis (RA) is a chronic inflammatory disease of the synovial joints causing pain deformities and disability. The highly vascular inflamed synovium has aggressive and destructive characteristics, it invades, erodes and gradually destroys cartilage and underlying bone. Photodynamic therapy (PDT) was performed using the chick chorioallantoic membrane (CAM) model to investigate the vitality of synovium and cartilage implanted on the CAM. Synovium, obtained from human patients, was grafted onto the CAM; gross microscopy and histology proved its vitality 7 days post grafting. Cartilage obtained from rabbit knee joint was also maintained on the CAM for 7 days. Its vitality was demonstrated by histology and by measuring metabolic and enzymatic activity of cartilage cells (chondrocytes) as well as the collagen and proteoglycans content. Selective PDT was performed using aluminum phthalocyanine tetrasulfonate (AlPcS4), a hydrophilic compound, soluble in biological solutions, as a photosensitizer. After irradiation with a diode laser (lambda equals 670 nm, 10 mW) damage was observed in vascularized synovium grafts, whereas avascular cartilage remained intact.

Inelastic behaviour of collagen networks in cell-matrix interactions and mechanosensation.

PubMed

Mohammadi, Hamid; Arora, Pamma D; Simmons, Craig A; Janmey, Paul A; McCulloch, Christopher A

2015-01-06

The mechanical properties of extracellular matrix proteins strongly influence cell-induced tension in the matrix, which in turn influences cell function. Despite progress on the impact of elastic behaviour of matrix proteins on cell-matrix interactions, little is known about the influence of inelastic behaviour, especially at the large and slow deformations that characterize cell-induced matrix remodelling. We found that collagen matrices exhibit deformation rate-dependent behaviour, which leads to a transition from pronounced elastic behaviour at fast deformations to substantially inelastic behaviour at slow deformations (1 μm min(-1), similar to cell-mediated deformation). With slow deformations, the inelastic behaviour of floating gels was sensitive to collagen concentration, whereas attached gels exhibited similar inelastic behaviour independent of collagen concentration. The presence of an underlying rigid support had a similar effect on cell-matrix interactions: cell-induced deformation and remodelling were similar on 1 or 3 mg ml(-1) attached collagen gels while deformations were two- to fourfold smaller in floating gels of high compared with low collagen concentration. In cross-linked collagen matrices, which did not exhibit inelastic behaviour, cells did not respond to the presence of the underlying rigid foundation. These data indicate that at the slow rates of collagen compaction generated by fibroblasts, the inelastic responses of collagen gels, which are influenced by collagen concentration and the presence of an underlying rigid foundation, are important determinants of cell-matrix interactions and mechanosensation. © 2014 The Author(s) Published by the Royal Society. All rights reserved.

MEMS deformable mirror embedded wavefront sensing and control system

NASA Astrophysics Data System (ADS)

Owens, Donald; Schoen, Michael; Bush, Keith

2006-01-01

Electrostatic Membrane Deformable Mirror (MDM) technology developed using silicon bulk micro-machining techniques offers the potential of providing low-cost, compact wavefront control systems for diverse optical system applications. Electrostatic mirror construction using bulk micro-machining allows for custom designs to satisfy wavefront control requirements for most optical systems. An electrostatic MDM consists of a thin membrane, generally with a thin metal or multi-layer high-reflectivity coating, suspended over an actuator pad array that is connected to a high-voltage driver. Voltages applied to the array elements deflect the membrane to provide an optical surface capable of correcting for measured optical aberrations in a given system. Electrostatic membrane DM designs are derived from well-known principles of membrane mechanics and electrostatics, the desired optical wavefront control requirements, and the current limitations of mirror fabrication and actuator drive electronics. MDM performance is strongly dependent on mirror diameter and air damping in meeting desired spatial and temporal frequency requirements. In this paper, we present wavefront control results from an embedded wavefront control system developed around a commercially available high-speed camera and an AgilOptics Unifi MDM driver using USB 2.0 communications and the Linux development environment. This new product, ClariFast TM, combines our previous Clarifi TM product offering into a faster more streamlined version dedicated strictly to Hartmann Wavefront sensing.

Vitamin C and E Supplements Enhance the Antioxidant Capacity of Erythrocytes Obtained from Aged Rats.

PubMed

Xiong, Yanlian; Xiong, Yanlei; Zhou, Shuai; Sun, Yanan; Zhao, Yuqi; Ren, Xiaotong; Zhang, Yingfang; Zhang, Naili

2017-04-01

The main purpose of the present study was to investigate the effects of vitamin C and E supplements on the antioxidant capacity of erythrocytes obtained from young and aged rats. Male Wistar rats aged 3 and 24 months were used. Vitamins C and E were injected at doses of 200 mg/kg (day) intraperitoneally in young and aged groups. The antioxidant capacity, oxidant stress parameters, and deformability of red blood cells collected from different age stages were evaluated. An in vitro oxidation system was constructed to explore the mechanisms of antioxidant capacity change in the vitamin treatment groups. Treatment with vitamins C and E can effectively restore the antioxidant capacity and deformability of red blood cells (RBCs) in aged rats. Under in vitro oxidative conditions, an age-dependent decline in the influx rate of L-cysteine was observed. This was significantly improved following treatment with vitamins C and E. We present evidence of an improvement in the antioxidant capacity of RBCs by treatment with vitamins C and E in aged rats. These observations also suggest that treatment with vitamins C and E improves glutathione synthesis by enhancing the influx rate of L-cysteine through the modification of membrane proteins and lipids.

[Research on electricity frequency property of blood].

PubMed

Hu, Maoqing; Huang, Hua; Yuan, Zirun; Chen, Huaiqing; Den, Lihua

2006-02-01

On the basis of our previous work, the electric frequency property of human blood in different components, in physiological state and in pathological state (diabetes) are tested and analyzed in the range of 1Hz-20MHz progressively. Among the different components of blood; the lowest electrical impedance is serum; the plasma and the whole blood gradually become larger, the blood corpuscle is the largest one. Otherwise, the negative phase of serum is the largest, the plasma and the whole blood are lower, and the blood corpuscle is the lowest. Here, the question is why the effect of the electric capacity of serum and plasma is the biggest in the condition of no cell and cell membrane; diabetes mellitus is an endocrine disorder in which blood changes obviously, the electric frequency property of the blood of diabetic patients changes markedly; the electrical impedance of blood decreases (more obviously with low frequency), the negative phase increases (more obviously with high frequency). These indicate that the increase of electric conductivity in diabetic patients' blood is due to electric capacitance conductivity that is related to the changes of cell membrane, deformation abilities and aggregation of RBC. Related experiments demonstrate again that with the progressing of research in the electric frequency property of blood, we may use the theory and method of electricity to examine some important characters of blood in a different way, and so to corroborate other tests and analyses.