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Sample records for actin-rich membrane protrusions

  1. Lgr4 and Lgr5 drive the formation of long actin-rich cytoneme-like membrane protrusions

    PubMed Central

    Snyder, Joshua C.; Rochelle, Lauren K.; Marion, Sébastien; Lyerly, H. Kim; Barak, Larry S.; Caron, Marc G.

    2015-01-01

    ABSTRACT Embryonic development and adult tissue homeostasis require precise information exchange between cells and their microenvironment to coordinate cell behavior. A specialized class of ultra-long actin-rich filopodia, termed cytonemes, provides one mechanism for this spatiotemporal regulation of extracellular cues. We provide here a mechanism whereby the stem-cell marker Lgr5, and its family member Lgr4, promote the formation of cytonemes. Lgr4- and Lgr5-induced cytonemes exceed lengths of 80 µm, are generated through stabilization of nascent filopodia from an underlying lamellipodial-like network and functionally provide a pipeline for the transit of signaling effectors. As proof-of-principle, we demonstrate that Lgr5-induced cytonemes act as conduits for cell signaling by demonstrating that the actin motor and filopodial cargo carrier protein myosin X (Myo10) and the G-protein-coupled receptor (GPCR) signaling effector β-arrestin-2 (Arrb2) transit into cytonemes. This work delineates a biological function for Lgr4 and Lgr5 and provides the rationale to fully investigate Lgr4 and Lgr5 function and cytonemes in mammalian stem cell and cancer stem cell behavior. PMID:25653388

  2. Effects of Plasma Membrane Cholesterol Level and Cytoskeleton F-Actin on Cell Protrusion Mechanics

    PubMed Central

    Khatibzadeh, Nima; Spector, Alexander A.; Brownell, William E.; Anvari, Bahman

    2013-01-01

    Protrusions are deformations that form at the surface of living cells during biological activities such as cell migration. Using combined optical tweezers and fluorescent microscopy, we quantified the mechanical properties of protrusions in adherent human embryonic kidney cells in response to application of an external force at the cell surface. The mechanical properties of protrusions were analyzed by obtaining the associated force-length plots during protrusion formation, and force relaxation at constant length. Protrusion mechanics were interpretable by a standard linear solid (Kelvin) model, consisting of two stiffness parameters, k0 and k1 (with k0>k1), and a viscous coefficient. While both stiffness parameters contribute to the time-dependant mechanical behavior of the protrusions, k0 and k1 in particular dominated the early and late stages of the protrusion formation and elongation process, respectively. Lowering the membrane cholesterol content by 25% increased the k0 stiffness by 74%, and shortened the protrusion length by almost half. Enhancement of membrane cholesterol content by nearly two-fold increased the protrusion length by 30%, and decreased the k0 stiffness by nearly two-and-half-fold as compared with control cells. Cytoskeleton integrity was found to make a major contribution to protrusion mechanics as evidenced by the effects of F-actin disruption on the resulting mechanical parameters. Viscoelastic behavior of protrusions was further characterized by hysteresis and force relaxation after formation. The results of this study elucidate the coordination of plasma membrane composition and cytoskeleton during protrusion formation. PMID:23451167

  3. Myosin-X facilitates Shigella-induced membrane protrusions and cell-to-cell spread

    PubMed Central

    Li, Wei; Dhillon, Jess; Bohil, Aparna B.; Cheney, Richard E.; Hartwig, John H.; Southwick, Frederick S.

    2014-01-01

    Summary The intracellular pathogen Shigella flexneri forms membrane protrusions to spread from cell to cell. As protrusions form, myosin-X (Myo10) localizes to Shigella. Electron micrographs of immunogold-labelled Shigella-infected HeLa cells reveal that Myo10 concentrates at the bases and along the sides of bacteria within membrane protrusions. Time-lapse video microscopy shows that a full-length Myo10 GFP-construct cycles along the sides of Shigella within the membrane protrusions as these structures progressively lengthen. RNAi knock-down of Myo10 is associated with shorter protrusions with thicker stalks, and causes a >80% decrease in confluent cell plaque formation. Myo10 also concentrates in membrane protrusions formed by another intracellular bacteria, Listeria, and knock-down of Myo10 also impairs Listeria plaque formation. In Cos7 cells (contain low concentrations of Myo10), the expression of full-length Myo10 nearly doubles Shigella-induced protrusion length, and lengthening requires the head domain, as well as the tail-PH domain, but not the FERM domain. The GFP-Myo10-HMM domain localizes to the sides of Shigella within membrane protrusions and the GFP-Myo10-PH domain localizes to host cell membranes. We conclude that Myo10 generates the force to enhance bacterial-induced protrusions by binding its head region to actin filaments and its PH tail domain to the peripheral membrane. PMID:23083060

  4. 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.

  5. Myosin motor function: the ins and outs of actin-based membrane protrusions

    PubMed Central

    Nambiar, Rajalakshmi; McConnell, Russell E.

    2011-01-01

    Cells build plasma membrane protrusions supported by parallel bundles of F-actin to enable a wide variety of biological functions, ranging from motility to host defense. Filopodia, microvilli and stereocilia are three such protrusions that have been the focus of intense biological and biophysical investigation in recent years. While it is evident that actin dynamics play a significant role in the formation of these organelles, members of the myosin superfamily have also been implicated as key players in the maintenance of protrusion architecture and function. Based on a simple analysis of the physical forces that control protrusion formation and morphology, as well as our review of available data, we propose that myosins play two general roles within these structures: (1) as cargo transporters to move critical regulatory components toward distal tips and (2) as mediators of membrane-cytoskeleton adhesion. PMID:20107861

  6. Normal mammary epithelial cells promote carcinoma basement membrane invasion by inducing microtubule-rich protrusions

    PubMed Central

    Lee, Meng-Horng; Wu, Pei-Hsun; Gilkes, Daniele; Aifuwa, Ivie; Wirtz, Denis

    2015-01-01

    Recent work suggests that the dissemination of tumor cells may occur in parallel with, and even preceed, tumor growth. The mechanism for this early invasion is largely unknown. Here, we find that mammary epithelial cells (MECs) induce neighboring breast carcinoma cells (BCCs) to cross the basement membrane by secreting soluble laminin. Laminin continuously produced by MECs induce long membrane cellular protrusions in BCCs that promote their contractility and invasion into the surrounding matrix. These protrusions depend on microtubule bundles assembled de novo through laminin-integrin β1 signaling. These results describe how non-cancerous MECs can actively participate in the invasive process of BCCs. PMID:26334095

  7. Fluxes of Water through Aquaporin 9 Weaken Membrane-Cytoskeleton Anchorage and Promote Formation of Membrane Protrusions

    PubMed Central

    Karlsson, Thommie; Bolshakova, Anastasia; Magalhães, Marco A. O.; Loitto, Vesa M.; Magnusson, Karl-Eric

    2013-01-01

    All modes of cell migration require rapid rearrangements of cell shape, allowing the cell to navigate within narrow spaces in an extracellular matrix. Thus, a highly flexible membrane and a dynamic cytoskeleton are crucial for rapid cell migration. Cytoskeleton dynamics and tension also play instrumental roles in the formation of different specialized cell membrane protrusions, viz. lamellipodia, filopodia, and membrane blebs. The flux of water through membrane-anchored water channels, known as aquaporins (AQPs) has recently been implicated in the regulation of cell motility, and here we provide novel evidence for the role of AQP9 in the development of various forms of membrane protrusion. Using multiple imaging techniques and cellular models we show that: (i) AQP9 induced and accumulated in filopodia, (ii) AQP9-associated filopodial extensions preceded actin polymerization, which was in turn crucial for their stability and dynamics, and (iii) minute, local reductions in osmolarity immediately initiated small dynamic bleb-like protrusions, the size of which correlated with the reduction in osmotic pressure. Based on this, we present a model for AQP9-induced membrane protrusion, where the interplay of water fluxes through AQP9 and actin dynamics regulate the cellular protrusive and motile activity of cells. PMID:23573219

  8. Membrane protrusion powers clathrin-independent endocytosis of interleukin-2 receptor.

    PubMed

    Basquin, Cyril; Trichet, Michaël; Vihinen, Helena; Malardé, Valérie; Lagache, Thibault; Ripoll, Léa; Jokitalo, Eija; Olivo-Marin, Jean-Christophe; Gautreau, Alexis; Sauvonnet, Nathalie

    2015-08-13

    Endocytosis controls many functions including nutrient uptake, cell division, migration and signal transduction. A clathrin- and caveolin-independent endocytosis pathway is used by important physiological cargos, including interleukin-2 receptors (IL-2R). However, this process lacks morphological and dynamic data. Our electron microscopy (EM) and tomography studies reveal that IL-2R-pits and vesicles are initiated at the base of protrusions. We identify the WAVE complex as a specific endocytic actor. The WAVE complex interacts with IL-2R, via a WAVE-interacting receptor sequence (WIRS) present in the receptor polypeptide, and allows for receptor clustering close to membrane protrusions. In addition, using total internal reflection fluorescent microscopy (TIRF) and automated analysis we demonstrate that two timely distinct bursts of actin polymerization are required during IL-2R uptake, promoted first by the WAVE complex and then by N-WASP. Finally, our data reveal that dynamin acts as a transition controller for the recruitment of Arp2/3 activators required for IL-2R endocytosis. Altogether, our work identifies the spatio-temporal specific role of factors initiating clathrin-independent endocytosis by a unique mechanism that does not depend on the deformation of a flat membrane, but rather on that of membrane protrusions.

  9. Dynamic Localization of G-actin During Membrane Protrusion in Neuronal Motility

    PubMed Central

    Lee, Chi Wai; Vitriol, Eric A.; Shim, Sangwoo; Wise, Ariel L.; Velayutham, Radhi P.; Zheng, James Q.

    2013-01-01

    Summary Background Actin-based cell motility is fundamental for the development, function, and malignant events of eukaryotic organisms. During neural development, axonal growth cones depend on rapid assembly and disassembly of actin filaments (F-actin) for their guided extension to specific targets for wiring. Monomeric globular actin (G-actin) is the building block for F-actin but is not considered to play a direct role in spatiotemporal control of actin dynamics in cell motility. Results Here we report that a pool of G-actin dynamically localizes to the leading edge of growth cones and neuroblastoma cells to spatially elevate the G-/F-actin ratio that drives membrane protrusion and cell movement. Loss of G-actin localization leads to the cessation and retraction of membrane protrusions. Moreover, G-actin localization occurs asymmetrically in growth cones during attractive turning. Finally, we identify the actin monomer binding proteins profilin and thymosin β4 as key molecules that localize actin monomers to the leading edge of lamellipodia for their motility. Conclusions Our results suggest that dynamic localization of G-actin provides a novel mechanism to regulate the spatiotemporal actin dynamics underlying membrane protrusion in cell locomotion and growth cone chemotaxis. PMID:23746641

  10. Membrane protrusion powers clathrin-independent endocytosis of interleukin-2 receptor

    PubMed Central

    Basquin, Cyril; Trichet, Michaël; Vihinen, Helena; Malardé, Valérie; Lagache, Thibault; Ripoll, Léa; Jokitalo, Eija; Olivo-Marin, Jean-Christophe; Gautreau, Alexis; Sauvonnet, Nathalie

    2015-01-01

    Endocytosis controls many functions including nutrient uptake, cell division, migration and signal transduction. A clathrin- and caveolin-independent endocytosis pathway is used by important physiological cargos, including interleukin-2 receptors (IL-2R). However, this process lacks morphological and dynamic data. Our electron microscopy (EM) and tomography studies reveal that IL-2R-pits and vesicles are initiated at the base of protrusions. We identify the WAVE complex as a specific endocytic actor. The WAVE complex interacts with IL-2R, via a WAVE-interacting receptor sequence (WIRS) present in the receptor polypeptide, and allows for receptor clustering close to membrane protrusions. In addition, using total internal reflection fluorescent microscopy (TIRF) and automated analysis we demonstrate that two timely distinct bursts of actin polymerization are required during IL-2R uptake, promoted first by the WAVE complex and then by N-WASP. Finally, our data reveal that dynamin acts as a transition controller for the recruitment of Arp2/3 activators required for IL-2R endocytosis. Altogether, our work identifies the spatio-temporal specific role of factors initiating clathrin-independent endocytosis by a unique mechanism that does not depend on the deformation of a flat membrane, but rather on that of membrane protrusions. PMID:26124312

  11. Mechanisms of leading edge protrusion in interstitial migration

    PubMed Central

    Wilson, Kerry; Lewalle, Alexandre; Fritzsche, Marco; Thorogate, Richard; Duke, Tom; Charras, Guillaume

    2013-01-01

    While the molecular and biophysical mechanisms underlying cell protrusion on two-dimensional substrates are well understood, our knowledge of the actin structures driving protrusion in three-dimensional environments is poor, despite relevance to inflammation, development and cancer. Here we report that, during chemotactic migration through microchannels with 5 μm × 5 μm cross-sections, HL60 neutrophil-like cells assemble an actin-rich slab filling the whole channel cross-section at their front. This leading edge comprises two distinct F-actin networks: an adherent network that polymerizes perpendicular to cell-wall interfaces and a ‘free’ network that grows from the free membrane at the cell front. Each network is polymerized by a distinct nucleator and, due to their geometrical arrangement, the networks interact mechanically. On the basis of our experimental data, we propose that, during interstitial migration, medial growth of the adherent network compresses the free network preventing its retrograde movement and enabling new polymerization to be converted into forward protrusion. PMID:24305616

  12. The F-BAR domain of srGAP2 induces membrane protrusions required for neuronal migration and morphogenesis

    PubMed Central

    Guerrier, Sabrice; Coutinho-Budd, Jaeda; Sassa, Takayuki; Gresset, Aurélie; Jordan, Nicole Vincent; Cheng, Ken; Jin, Wei-Lin; Frost, Adam; Polleux, Franck

    2009-01-01

    SUMMARY During brain development, proper neuronal migration and morphogenesis is critical for the establishment of functional neural circuits. Here we report that srGAP2 negatively regulates neuronal migration and induces neurite outgrowth and branching through the ability of its F-BAR domain to induce filopodia-like membrane protrusions resembling those induced by I-BAR domains in vivo and in vitro. Previous work has suggested that in non-neuronal cells, filopodia dynamics decreases the rate of cell migration and the persistence of leading edge protrusions. srGAP2 knockdown reduces leading process branching and increases the rate of neuronal migration in vivo. Overexpression of srGAP2 or its F-BAR domain has the opposite effects, increasing leading process branching and decreasing migration. These results (1) suggest that F-BAR domains are functionally diverse and (2) highlight the functional importance of proteins directly regulating membrane deformation for proper neuronal migration and morphogenesis. PMID:19737524

  13. Tetraspanin CD82 Inhibits Protrusion and Retraction in Cell Movement by Attenuating the Plasma Membrane-Dependent Actin Organization

    PubMed Central

    Liu, Wei M.; Zhang, Feng; Moshiach, Simon; Zhou, Bin; Huang, Chao; Srinivasan, Kamalakkannan; Khurana, Seema; Zheng, Yi; Lahti, Jill M.; Zhang, Xin A.

    2012-01-01

    To determine how tetraspanin KAI1/CD82, a tumor metastasis suppressor, inhibits cell migration, we assessed which cellular events critical for motility are altered by KAI1/CD82 and how KAI1/CD82 regulates these events. We found that KAI1/CD82-expressing cells typically exhibited elongated cellular tails and diminished lamellipodia. Live imaging demonstrated that the polarized protrusion and retraction of the plasma membrane became deficient upon KAI1/CD82 expression. The deficiency in developing these motility-related cellular events was caused by poor formations of actin cortical network and stress fiber and by aberrant dynamics in actin organization. Rac1 activity was reduced by KAI1/CD82, consistent with the diminution of lamellipodia and actin cortical network; while the growth factor-stimulated RhoA activity was blocked by KAI1/CD82, consistent with the loss of stress fiber and attenuation in cellular retraction. Upon KAI1/CD82 expression, Rac effector cofilin was not enriched at the cell periphery to facilitate lamellipodia formation while Rho kinase exhibited a significantly lower activity leading to less retraction. Phosphatidylinositol 4, 5-biphosphate, which initiates actin polymerization from the plasma membrane, became less detectable at the cell periphery in KAI1/CD82-expressing cells. Moreover, KAI1/CD82-induced phenotypes likely resulted from the suppression of multiple signaling pathways such as integrin and growth factor signaling. In summary, at the cellular level KAI1/CD82 inhibited polarized protrusion and retraction events by disrupting actin reorganization; at the molecular level, KAI1/CD82 deregulated Rac1, RhoA, and their effectors cofilin and Rho kinase by perturbing the plasma membrane lipids. PMID:23251627

  14. Protrusion force microscopy reveals oscillatory force generation and mechanosensing activity of human macrophage podosomes

    NASA Astrophysics Data System (ADS)

    Labernadie, Anna; Bouissou, Anaïs; Delobelle, Patrick; Balor, Stéphanie; Voituriez, Raphael; Proag, Amsha; Fourquaux, Isabelle; Thibault, Christophe; Vieu, Christophe; Poincloux, Renaud; Charrière, Guillaume M.; Maridonneau-Parini, Isabelle

    2014-11-01

    Podosomes are adhesion structures formed in monocyte-derived cells. They are F-actin-rich columns perpendicular to the substrate surrounded by a ring of integrins. Here, to measure podosome protrusive forces, we designed an innovative experimental setup named protrusion force microscopy (PFM), which consists in measuring by atomic force microscopy the deformation induced by living cells onto a compliant Formvar sheet. By quantifying the heights of protrusions made by podosomes onto Formvar sheets, we estimate that a single podosome generates a protrusion force that increases with the stiffness of the substratum, which is a hallmark of mechanosensing activity. We show that the protrusive force generated at podosomes oscillates with a constant period and requires combined actomyosin contraction and actin polymerization. Finally, we elaborate a model to explain the mechanical and oscillatory activities of podosomes. Thus, PFM shows that podosomes are mechanosensing cell structures exerting a protrusive force.

  15. Protrusion force microscopy reveals oscillatory force generation and mechanosensing activity of human macrophage podosomes.

    PubMed

    Labernadie, Anna; Bouissou, Anaïs; Delobelle, Patrick; Balor, Stéphanie; Voituriez, Raphael; Proag, Amsha; Fourquaux, Isabelle; Thibault, Christophe; Vieu, Christophe; Poincloux, Renaud; Charrière, Guillaume M; Maridonneau-Parini, Isabelle

    2014-01-01

    Podosomes are adhesion structures formed in monocyte-derived cells. They are F-actin-rich columns perpendicular to the substrate surrounded by a ring of integrins. Here, to measure podosome protrusive forces, we designed an innovative experimental setup named protrusion force microscopy (PFM), which consists in measuring by atomic force microscopy the deformation induced by living cells onto a compliant Formvar sheet. By quantifying the heights of protrusions made by podosomes onto Formvar sheets, we estimate that a single podosome generates a protrusion force that increases with the stiffness of the substratum, which is a hallmark of mechanosensing activity. We show that the protrusive force generated at podosomes oscillates with a constant period and requires combined actomyosin contraction and actin polymerization. Finally, we elaborate a model to explain the mechanical and oscillatory activities of podosomes. Thus, PFM shows that podosomes are mechanosensing cell structures exerting a protrusive force. PMID:25385672

  16. Arp2/3 Inhibition Induces Amoeboid-Like Protrusions in MCF10A Epithelial Cells by Reduced Cytoskeletal-Membrane Coupling and Focal Adhesion Assembly

    PubMed Central

    Stricker, Jonathan; Sayegh, Kareem; Campillo, Clement; Gardel, Margaret L.

    2014-01-01

    Here we demonstrate that Arp2/3 regulates a transition between mesenchymal and amoeboid protrusions in MCF10A epithelial cells. Using genetic and pharmacological means, we first show Arp2/3 inhibition impairs directed cell migration. Arp2/3 inhibition results in a dramatically impaired cell adhesion, causing deficient cell attachment and spreading to ECM as well as an 8-fold decrease in nascent adhesion assembly at the leading edge. While Arp2/3 does not play a significant role in myosin-dependent adhesion growth, mature focal adhesions undergo large scale movements against the ECM suggesting reduced coupling to the ECM. Cell edge protrusions occur at similar rates when Arp2/3 is inhibited but their morphology is dramatically altered. Persistent lamellipodia are abrogated and we observe a markedly increased incidence of blebbing and unstable pseuodopods. Micropipette-aspiration assays indicate that Arp2/3-inhibited cells have a weak coupling between the cell cortex and the plasma membrane, and suggest a potential mechanism for increased pseudopod and bleb formation. Pseudopods are not sensitive to reduced in formin or myosin II activity. Collectively, these results indicate that Arp2/3 is not necessary for rapid protrusion of the cell edge but plays a crucial role in assembling focal adhesions required for its stabilization. PMID:24967897

  17. Dynamic Membrane Remodeling at Invadopodia Differentiates Invadopodia from Podosomes

    PubMed Central

    Artym, Vira V.; Matsumoto, Kazue; Mueller, Susette C.; Yamada, Kenneth M.

    2011-01-01

    Invadopodia are specialized actin-rich protrusions of metastatic tumor and transformed cells with crucial functions in ECM degradation and invasion. Although early electron microscopy studies described invadopodia as long filament-like protrusions of the cell membrane adherent to the matrix, fluorescence microscopy studies have focused on invadopodia as actin-cortactin aggregates localized to areas of ECM degradation. The absence of a clear conceptual integration of these two descriptions of invadopodial structure has impeded understanding of the regulatory mechanisms that govern invadopodia. To determine the relationship between the membrane filaments identified by electron microscopy and the actin-cortactin aggregates of invadopodia, we applied rapid live-cell high-resolution TIRF microscopy to examine cell membrane dynamics at the cortactin core of the invadopodia of human carcinoma cells. We found that cortactin docking to the cell membrane adherent to 2D fibronectin matrix initiates invadopodium assembly associated with the formation of a invadopodial membrane process that extends from a ventral cell membrane lacuna toward the ECM. The tip of the invadopodial process flattens as it interacts with the 2D matrix, and it undergoes constant rapid ruffling and dynamic formation of filament-like protrusions as the invadopodium matures. To describe this newly discovered dynamic relationship between the actin-cortactin core and invadopodial membranes, we propose a model of the invadopodial complex. Using TIRF microscopy, we also established that – in striking contrast to the invadopodium – membrane at the podosome of a macrophage fails to form any process- or filament-like membrane protrusions. Thus, the undulation and ruffling of the invadopodial membrane together with the formation of dynamic filament-like extensions from the invadopodial cortactin core defines invadopodia as invasive superstructures that are distinct from the podosomes. PMID:20656375

  18. Endothelial Surface Protrusion by a Point Force.

    PubMed

    Chen, Yong; Lu, Lan; Shao, Jin-Yu

    2016-03-01

    During leukocyte rolling on the endothelium, surface protrusion and membrane tether extraction occur consecutively on leukocytes. Both surface protrusion and tether extraction of leukocytes stabilize leukocyte rolling. Tethers can also be extracted from endothelial cells (ECs), but surface protrusion of ECs has never been confirmed to exist. In this study, we examined EC surface protrusion with the micropipette aspiration technique. We found that, like leukocytes, surface protrusion on an EC did exist when a point force was imposed. Both the protrusional stiffness and the crossover force of EC surface protrusion were dependent on the force loading rate and the cytoskeletal integrity, but neither of them was dependent on tumor necrosis factor α stimulation. Temperature (37°C) affected the protrusional stiffness only at small force loading rates. When a neutrophil was employed to directly impose the pulling force on the EC, simultaneous surface protrusion from both cells occurred, and it can be modeled as two springs connected in series, although the spring constants should be adjusted according to the force loading rate. Therefore, EC surface protrusion is an important aspect of leukocyte rolling, and it should not be ignored when leukocyte rolling stability is studied systematically.

  19. Myosin Va facilitates the distribution of secretory granules in the F-actin rich cortex of PC12 cells.

    PubMed

    Rudolf, Rüdiger; Kögel, Tanja; Kuznetsov, Sergei A; Salm, Thorsten; Schlicker, Oliver; Hellwig, Andrea; Hammer, John A; Gerdes, Hans-Hermann

    2003-04-01

    Neuroendocrine secretory granules, the storage organelles for neuropeptides and hormones, are formed at the trans-Golgi network, stored inside the cell and exocytosed upon stimulation. Previously, we have reported that newly formed secretory granules of PC12 cells are transported in a microtubule-dependent manner from the trans-Golgi network to the F-actin-rich cell cortex, where they undergo short directed movements and exhibit a homogeneous distribution. Here we provide morphological and biochemical evidence that myosin Va is associated with secretory granules. Expression of a dominant-negative tail domain of myosin Va in PC12 cells led to an extensive clustering of secretory granules close to the cell periphery, a loss of their cortical restriction and a strong reduction in their motility in the actin cortex. Based on this data we propose a model that implies a dual transport system for secretory granules: after microtubule-dependent delivery to the cell periphery, secretory granules exhibit a myosin Va-dependent transport leading to their restriction and even dispersal in the F-actin-rich cortex of PC12 cells. PMID:12615975

  20. The initiation of post-synaptic protrusions

    PubMed Central

    Hotulainen, Pirta; Saarikangas, Juha

    2016-01-01

    ABSTRACT The post-synaptic spines of neuronal dendrites are highly elaborate membrane protrusions. Their anatomy, stability and density are intimately linked to cognitive performance. The morphological transitions of spines are powered by coordinated polymerization of actin filaments against the plasma membrane, but how the membrane-associated polymerization is spatially and temporally regulated has remained ill defined. Here, we discuss our recent findings showing that dendritic spines can be initiated by direct membrane bending by the I-BAR protein MIM/Mtss1. This lipid phosphatidylinositol (PI(4,5)P2) signaling-activated membrane bending coordinated spatial actin assembly and promoted spine formation. From recent advances, we formulate a general model to discuss how spatially concentrated protein-lipid microdomains formed by multivalent interactions between lipids and actin/membrane regulatory proteins might launch cell protrusions. PMID:27489575

  1. The mechanics of cell protrusion

    NASA Astrophysics Data System (ADS)

    Danuser, Gaudenz

    2006-03-01

    The protrusion of the cell edge is the first step in a cycle of molecular processes that drive cell movements during development, immune responses, wound healing and many other physiological functions. It is also the earliest pathological event observed during metastasis of cancer. Textbook models associate protrusion with the assembly of an actin polymer network subadjacent to the cell plasma membrane. However, for this process to be transformed into edge advancement, polymerization-induced forces need to be balanced by adhesion complexes that link the actin network to the extracellular domain. Also, the effectiveness of network assembly in mediating forward movement of the cell edge depends on how contraction forces pull the network in the cell front retrogradly towards the cell center. Thus, what is observed in a microscope as cell protrusion reflects the kinematic output of at least three space- and time-modulated mechanisms of force generation. The coordination of these machineries is thought to be regulated by a complex network of mechano-chemical signals. Our goal is to establish the contributions of each those mechanisms and their control by reconstructing the spatiotemporal distribution of intracellular forces via inverse dynamics and molecular intervention with the relevant signalling pathways. To this end, we have developed quantitative Fluorescent Speckle Microscopy (qFSM) which provides high-resolution spatiotemporal measurements of actin network deformation and material properties in migrating cells. In addition, qFSM delivers maps of cytoskeleton assembly and disassembly, so that we can infer the plasticity of the material in situ. Together, this data allows us to deduce intracellular force distributions from the constitutive laws of strain and stress in the actin polymer network. Using this approach we discovered that unperturbed cells protrude in a dynamic steady state where periodic patterns of network assembly, adhesion formation, and

  2. Cell Protrusions and Tethers: A Unified Approach

    PubMed Central

    Pospieszalska, Maria K.; Lasiecka, Irena; Ley, Klaus

    2011-01-01

    Low pulling forces applied locally to cell surface membranes produce viscoelastic cell surface protrusions. As the force increases, the membrane can locally separate from the cytoskeleton and a tether forms. Tethers can grow to great lengths exceeding the cell diameter. The protrusion-to-tether transition is known as the crossover. Here we propose a unified approach to protrusions and tethers providing, to our knowledge, new insights into their biomechanics. We derive a necessary and sufficient condition for a crossover to occur, a formula for predicting the crossover time, conditions for a tether to establish a dynamic equilibrium (characterized by constant nonzero pulling force and tether extension rate), a general formula for the tether material after crossover, and a general modeling method for tether pulling experiments. We introduce two general protrusion parameters, the spring constant and effective viscosity, valid before and after crossover. Their first estimates for neutrophils are 50 pN μm−1 and 9 pN s μm−1, respectively. The tether elongation after crossover is described as elongation of a viscoelastic-like material with a nonlinearly decaying spring (NLDs-viscoelastic material). Our model correctly describes the results of the published protrusion and tether pulling experiments, suggesting that it is universally applicable to such experiments. PMID:21463583

  3. Powered protrusion cutter

    DOEpatents

    Bzorgi, Fariborz M.

    2010-03-09

    An apparatus for clipping a protrusion of material is provided. The protrusion may, for example, be a bolt head, a nut, a rivet, a weld bead, or a temporary assembly alignment tab protruding from a substrate surface of assembled components. The apparatus typically includes a cleaver having a cleaving edge and a cutting blade having a cutting edge. Generally, a mounting structure configured to confine the cleaver and the cutting blade and permit a range of relative movement between the cleaving edge and the cutting edge is provided. Also typically included is a power device coupled to the cutting blade. The power device is configured to move the cutting edge toward the cleaving edge. In some embodiments the power device is activated by a momentary switch. A retraction device is also generally provided, where the retraction device is configured to move the cutting edge away from the cleaving edge.

  4. Device for cutting protrusions

    DOEpatents

    Bzorgi, Fariborz M.

    2011-07-05

    An apparatus for clipping a protrusion of material is provided. The protrusion may, for example, be a bolt head, a nut, a rivet, a weld bead, or a temporary assembly alignment tab protruding from a substrate surface of assembled components. The apparatus typically includes a cleaver having a cleaving edge and a cutting blade having a cutting edge. Generally, a mounting structure configured to confine the cleaver and the cutting blade and permit a range of relative movement between the cleaving edge and the cutting edge is provided. Also typically included is a power device coupled to the cutting blade. The power device is configured to move the cutting edge toward the cleaving edge. In some embodiments the power device is activated by a momentary switch. A retraction device is also generally provided, where the retraction device is configured to move the cutting edge away from the cleaving edge.

  5. Contact angle at the leading edge controls cell protrusion rate.

    PubMed

    Gabella, Chiara; Bertseva, Elena; Bottier, Céline; Piacentini, Niccolò; Bornert, Alicia; Jeney, Sylvia; Forró, László; Sbalzarini, Ivo F; Meister, Jean-Jacques; Verkhovsky, Alexander B

    2014-05-19

    Plasma membrane tension and the pressure generated by actin polymerization are two antagonistic forces believed to define the protrusion rate at the leading edge of migrating cells [1-5]. Quantitatively, resistance to actin protrusion is a product of membrane tension and mean local curvature (Laplace's law); thus, it depends on the local geometry of the membrane interface. However, the role of the geometry of the leading edge in protrusion control has not been yet investigated. Here, we manipulate both the cell shape and substrate topography in the model system of persistently migrating fish epidermal keratocytes. We find that the protrusion rate does not correlate with membrane tension, but, instead, strongly correlates with cell roundness, and that the leading edge of the cell exhibits pinning on substrate ridges-a phenomenon characteristic of spreading of liquid drops. These results indicate that the leading edge could be considered a triple interface between the substrate, membrane, and extracellular medium and that the contact angle between the membrane and the substrate determines the load on actin polymerization and, therefore, the protrusion rate. Our findings thus illuminate a novel relationship between the 3D shape of the cell and its dynamics, which may have implications for cell migration in 3D environments.

  6. Protrusion Fluctuations Direct Cell Motion

    PubMed Central

    Caballero, David; Voituriez, Raphaël; Riveline, Daniel

    2014-01-01

    Many physiological phenomena involve directional cell migration. It is usually attributed to chemical gradients in vivo. Recently, other cues have been shown to guide cells in vitro, including stiffness/adhesion gradients or micropatterned adhesive motifs. However, the cellular mechanism leading to these biased migrations remains unknown, and, often, even the direction of motion is unpredictable. In this study, we show the key role of fluctuating protrusions on ratchet-like structures in driving NIH3T3 cell migration. We identified the concept of efficient protrusion and an associated direction index. Our analysis of the protrusion statistics facilitated the quantitative prediction of cell trajectories in all investigated conditions. We varied the external cues by changing the adhesive patterns. We also modified the internal cues using drug treatments, which modified the protrusion activity. Stochasticity affects the short- and long-term steps. We developed a theoretical model showing that an asymmetry in the protrusion fluctuations is sufficient for predicting all measures associated with the long-term motion, which can be described as a biased persistent random walk. PMID:24988339

  7. 16 CFR 1511.4 - Protrusions.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... orientation of the hinge axis shall be horizontal. A plane surface shall be applied to any protrusion from the... axis of the nipple. Any protrusion shall be allowed to flex or rotate about its hinge as the...

  8. The Calponin Family Member CHDP-1 Interacts with Rac/CED-10 to Promote Cell Protrusions

    PubMed Central

    Zhang, Jingyan; Liu, Jia-Jia; Wang, Yingchun; Ding, Mei

    2016-01-01

    Eukaryotic cells extend a variety of surface protrusions to direct cell motility. Formation of protrusions is mediated by coordinated actions between the plasma membrane and the underlying actin cytoskeleton. Here, we found that the single calponin homology (CH) domain-containing protein CHDP-1 induces the formation of cell protrusions in C. elegans. CHDP-1 is anchored to the cortex through its amphipathic helix. CHDP-1 associates through its CH domain with the small GTPase Rac1/CED-10, which is a key regulator of the actin cytoskeleton. CHDP-1 preferentially binds to the GTP-bound active form of the CED-10 protein and preserves the membrane localization of GTP-CED-10. Hence, by coupling membrane expansion to Rac1-mediated actin dynamics, CHDP-1 promotes the formation of cellular protrusions in vivo. PMID:27415421

  9. The Calponin Family Member CHDP-1 Interacts with Rac/CED-10 to Promote Cell Protrusions.

    PubMed

    Guan, Liying; Ma, Xuehua; Zhang, Jingyan; Liu, Jia-Jia; Wang, Yingchun; Ding, Mei

    2016-07-01

    Eukaryotic cells extend a variety of surface protrusions to direct cell motility. Formation of protrusions is mediated by coordinated actions between the plasma membrane and the underlying actin cytoskeleton. Here, we found that the single calponin homology (CH) domain-containing protein CHDP-1 induces the formation of cell protrusions in C. elegans. CHDP-1 is anchored to the cortex through its amphipathic helix. CHDP-1 associates through its CH domain with the small GTPase Rac1/CED-10, which is a key regulator of the actin cytoskeleton. CHDP-1 preferentially binds to the GTP-bound active form of the CED-10 protein and preserves the membrane localization of GTP-CED-10. Hence, by coupling membrane expansion to Rac1-mediated actin dynamics, CHDP-1 promotes the formation of cellular protrusions in vivo. PMID:27415421

  10. Actin Grips: Circular Actin-Rich Cytoskeletal Structures that Mediate the Wrapping of Polymeric Microfibers by Endothelial Cells

    PubMed Central

    Jones, Desiree; Park, DoYoung; Anghelina, Mirela; Pecot, Thierry; Machiraju, Raghu; Xue, Ruipeng; Lannutti, John; Thomas, Jessica; Cole, Sara; Moldovan, Leni; Moldovan, Nicanor I.

    2015-01-01

    Interaction of endothelial-lineage cells with three-dimensional substrates was much less studied than that with flat culture surfaces. We investigated the in vitro attachment of both mature endothelial cells (ECs) and of less differentiated EC colony-forming cells to poly-e-capro-lactone (PCL) fibers with diameters in 5–20 μm range (‘scaffold microfibers’, SMFs). We found that notwithstanding the poor intrinsic adhesiveness to PCL, both cell types completely wrapped the SMFs after long-term cultivation, thus attaining a cylindrical morphology. In this system, both EC types grew vigorously for more than a week and became increasingly more differentiated, as shown by multiplexed gene expression. Three-dimensional reconstructions from multiphoton confocal microscopy images using custom software showed that the filamentous (F) actin bundles took a conspicuous ring-like organization around the SMFs. Unlike the classical F-actin-containing stress fibers, these rings were not associated with either focal adhesions or intermediate filaments. We also demonstrated that plasma membrane boundaries adjacent to these circular cytoskeletal structures were tightly yet dynamically apposed to the SMFs, for which reason we suggest to call them ‘actin grips’. In conclusion, we describe a particular form of F-actin assembly with relevance for cytoskeletal organization in response to biomaterials, for endothelial-specific cell behavior in vitro and in vivo, and for tissue engineering. PMID:25818446

  11. Actin is not required for nanotubular protrusions of primary astrocytes grown on metal nano-lawn.

    PubMed

    Gimsa, Ulrike; Iglic, Ales; Fiedler, Stefan; Zwanzig, Michael; Kralj-Iglic, Veronika; Jonas, Ludwig; Gimsa, Jan

    2007-01-01

    We used sub-micron metal rod decorated surfaces, 'nano-lawn' structures, as a substrate to study cell-to-cell and cell-to-surface interactions of primary murine astrocytes. These cells form thin membranous tubes with diameters of less than 100 nm and a length of several microns, which make contact to neighboring cells and the substrate during differentiation. While membrane protrusions grow on top of the nano-lawn pillars, nuclei sink to the bottom of the substrate. We observed gondola-like structures along those tubes, suggestive of their function as transport vehicles. Elements of the cytoskeleton such as actin fibers are commonly believed to be essential for triggering the onset and growth of tubular membrane protrusions. A rope-pulling mechanism along actin fibers has recently been proposed to account for the transport or exchange of cellular material between cells. We present evidence for a complementary mechanism that promotes growth and stabilization of the observed tubular protrusions of cell membranes. This mechanism does not require active involvement of actin fibers as the formation of membrane protrusions could not be prevented by suppressing polymerization of actin by latrunculin B. Also theoretically, actin fibers are not essential for the growing and stability of nanotubes since curvature-driven self-assembly of interacting anisotropic raft elements is sufficient for the spontaneous formation of thin nano-tubular membrane protrusions. PMID:17520481

  12. 16 CFR 1511.4 - Protrusions.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... orientation of the hinge axis shall be horizontal. A plane surface shall be applied to any protrusion from the... direction along the axis of the nipple. The normal of the plane surface shall be maintained parallel to the axis of the nipple. Any protrusion shall be allowed to flex or rotate about its hinge as the...

  13. Electrical assembly having heat sink protrusions

    DOEpatents

    Rinehart, Lawrence E.; Romero, Guillermo L.

    2009-04-21

    An electrical assembly, comprising a heat producing semiconductor device supported on a first major surface of a direct bond metal substrate that has a set of heat sink protrusions supported by its second major surface. In one preferred embodiment the heat sink protrusions are made of the same metal as is used in the direct bond copper.

  14. 16 CFR 1511.4 - Protrusions.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... orientation of the hinge axis shall be horizontal. A plane surface shall be applied to any protrusion from the... direction along the axis of the nipple. The normal of the plane surface shall be maintained parallel to the axis of the nipple. Any protrusion shall be allowed to flex or rotate about its hinge as the...

  15. Filopodia and Viruses: An Analysis of Membrane Processes in Entry Mechanisms

    PubMed Central

    Chang, Kenneth; Baginski, John; Hassan, Samer F.; Volin, Michael; Shukla, Deepak; Tiwari, Vaibhav

    2016-01-01

    Filopodia are thin, actin rich bundles protruding from cell plasma membranes, serving physiological purposes, such as probing the environment and facilitating cell-to-cell adhesion. Recent studies have highlighted that actively polymerized filopodial-protrusions are exploited during virus entry, trafficking, spread, and the development of clinical pathology of viral diseases. These observations have caused a surge in investigation of the key determinants of filopodial induction and their influence on cell topography including receptor expression for viral entry. It is now very clear that filopodia can provide unique opportunities for many viruses to invade host cells vertically during primary infection, or horizontally during virus spread from cell-to-cell. These emerging concepts can explain the unprecedented ability of viruses to invade both nearby and long-distant host cells, a feature that may directly contribute to viral tropism. In this review, we summarize the significance of filopodia in viral diseases and discuss future therapeutic possibilities to precisely target filopodial-flyovers to prevent or control infectious diseases. PMID:27014223

  16. Trespassing cancer cells: ‘fingerprinting’ invasive protrusions reveals metastatic culprits

    PubMed Central

    Klemke, Richard L.

    2012-01-01

    Metastatic cancer cells produce invasive membrane protrusions called invadopodia and pseudopodia, which play a central role in driving cancer cell dissemination in the body. Malignant cells use these structures to attach to and degrade extracellular matrix proteins, generate force for cell locomotion, and to penetrate the vasculature. Recent work using unique subcellular fractionation methodologies combined with spatial genomic, proteomic, and phosphoproteomic profiling has provided insight into the invadopodiome and pseudopodiome signaling networks that control the protrusion of invasive membranes. Here I highlight how these powerful spatial “omics” approaches reveal important signatures of metastatic cancer cells and possible new therapeutic targets aimed at treating metastatic disease. PMID:22980730

  17. Cell protrusions induced by hyaluronan synthase 3 (HAS3) resemble mesothelial microvilli and share cytoskeletal features of filopodia.

    PubMed

    Koistinen, Ville; Kärnä, Riikka; Koistinen, Arto; Arjonen, Antti; Tammi, Markku; Rilla, Kirsi

    2015-10-01

    Previous studies have shown that overexpression of enzymatically active GFP-HAS induces the growth of long, slender protrusions that share many features of both filopodia and microvilli. These protrusions are dependent on continuing hyaluronan synthesis, and disrupt upon digestion of hyaluronan by hyaluronidase. However, complete understanding of their nature is still missing. This work shows that the protrusions on rat peritoneal surface are ultrastructurally indistinguishable from those induced by GFP-HAS3 in MCF-7 cells. Analysis of the actin-associated proteins villin, ezrin, espin, fascin, and Myo10 indicated that the HAS3-induced protrusions share most cytoskeletal features with filopodia, but they do not require adherence to the substratum like traditional filopodia. GFP-HAS3 overexpression was found to markedly enhance filamentous actin in the protrusions and their cortical basis. Analysis of the protrusion dynamics after enzymatic digestion of hyaluronan revealed that while GFP-HAS3 escape from the protrusions and the protrusion collapse takes place immediately, the complete retraction of the protrusions occurs more slowly. This finding also suggests that hyaluronan chain maintains HAS3 in the plasma membrane. The results of this work suggest that protrusions similar to those of HAS3 overexpressing cells in vitro exist also in cells with active hyaluronan synthesis in vivo. These protrusions are similar to common filopodia but are independent of substratum attachment due to the extracellular scaffolding by the hyaluronan coat that accounts for the growth and maintenance of these structures, previously associated to invasion, adhesion and multidrug resistance. PMID:26162854

  18. Subcellular and Dynamic Coordination between Src Activity and Cell Protrusion in Microenvironment

    PubMed Central

    Zhuo, Yue; Qian, Tongcheng; Wu, Yiqian; Seong, Jihye; Gong, Ya; Ma, Hongwei; Wang, Yingxiao; Lu, Shaoying

    2015-01-01

    Migration of endothelial cells is essential for wound healing and angiogenesis. Src kinase activity plays important roles at the protrusions of migrating endothelial cells. However, the spatiotemporal coordination between Src kinase activity and the protrusion of cell edge remains unclear. Therefore, we investigate these coordinated molecular events at the initiation of cell migration, by integrating microfabrication, fluorescence resonance energy transfer (FRET)-based biosensors, and automated computational image analysis. We demonstrate that the physical release of restrictive micropattern triggered a significant decrease of Src activity at the protrusive edge of endothelial cells. Computational cross-correlation analysis reveals that the decrease of Src activity occurred earlier in time, and was well-coordinated with the protrusion of cell edge in polarized cells, but not in non-polarized cells. These results suggest that the spatiotemporal control of Src kinase activity is well-coordinated with cell polarization and protrusion in endothelial cells upon the release of physical constraint, as that experienced by endothelial cells sprouting from stiff tumor micro-environment during angiogenesis. Therefore, our integrative approach enabled the discovery of a new model where Src is de-activated in coordination with membrane protrusion, providing important insights into the regulation of endothelial migration and angiogenesis. PMID:26261043

  19. Polarized Cdc42 activation promotes polar body protrusion and asymmetric division in mouse oocytes

    PubMed Central

    Dehapiot, Benoit; Carrière, Virginie; Carroll, John; Halet, Guillaume

    2013-01-01

    Asymmetric meiotic divisions in mammalian oocytes rely on the eccentric positioning of the spindle and the remodeling of the overlying cortex, resulting in the formation of small polar bodies. The mechanism of this cortical polarization, exemplified by the formation of a thick F-actin cap, is poorly understood. Cdc42 is a major player in cell polarization in many systems; however, the spatio-temporal dynamics of Cdc42 activation during oocyte meiosis, and its contribution to mammalian oocyte polarization, have remained elusive. In this study, we investigated Cdc42 activation (Cdc42–GTP), dynamics and role during mouse oocyte meiotic divisions. We show that Cdc42–GTP accumulates in restricted cortical regions overlying meiotic chromosomes or chromatids, in a Ran–GTP-dependent manner. This polarized activation of Cdc42 is required for the recruitment of N-WASP and the formation of F-actin-rich protrusions during polar body formation. Cdc42 inhibition in MII oocytes resulted in the release of N-WASP into the cytosol, a loss of the polarized F-actin cap, and a failure to protrude the second polar body. Cdc42 inhibition also resulted in central spindle defects in activated MII oocytes. In contrast, emission of the first polar body during oocyte maturation could occur in the absence of a functional Cdc42/N-WASP pathway. Therefore, Cdc42 is a new protagonist in chromatin-induced cortical polarization in mammalian oocytes, with an essential role in meiosis II completion, through the recruitment and activation of N-WASP, downstream of the chromatin-centered Ran–GTP gradient. PMID:23384564

  20. Prominin-2 expression increases protrusions, decreases caveolae and inhibits Cdc42 dependent fluid phase endocytosis

    SciTech Connect

    Singh, Raman Deep Schroeder, Andreas S.; Scheffer, Luana; Holicky, Eileen L.; Wheatley, Christine L.; Marks, David L. Pagano, Richard E.

    2013-05-10

    Highlights: •Prominin-2 expression induced protrusions that co-localized with lipid raft markers. •Prominin-2 expression decreased caveolae, caveolar endocytosis and increased pCav1. •Prominin-2 expression inhibited fluid phase endocytosis by inactivation of Cdc42. •These endocytic effects can be reversed by adding exogenous cholesterol. •Caveolin1 knockdown restored fluid phase endocytosis in Prominin2 expressing cells. -- Abstract: Background: Membrane protrusions play important roles in biological processes such as cell adhesion, wound healing, migration, and sensing of the external environment. Cell protrusions are a subtype of membrane microdomains composed of cholesterol and sphingolipids, and can be disrupted by cholesterol depletion. Prominins are pentaspan membrane proteins that bind cholesterol and localize to plasma membrane (PM) protrusions. Prominin-1 is of great interest as a marker for stem and cancer cells, while Prominin-2 (Prom2) is reportedly restricted to epithelial cells. Aim: To characterize the effects of Prom-2 expression on PM microdomain organization. Methods: Prom2-fluorescent protein was transfected in human skin fibroblasts (HSF) and Chinese hamster ovary (CHO) cells for PM raft and endocytic studies. Caveolae at PM were visualized using transmission electron microscopy. Cdc42 activation was measured and caveolin-1 knockdown was performed using siRNAs. Results: Prom2 expression in HSF and CHO cells caused extensive Prom2-positive protrusions that co-localized with lipid raft markers. Prom2 expression significantly decreased caveolae at the PM, reduced caveolar endocytosis and increased caveolin-1 phosphorylation. Prom2 expression also inhibited Cdc42-dependent fluid phase endocytosis via decreased Cdc42 activation. Effects on endocytosis were reversed by addition of cholesterol. Knockdown of caveolin-1 by siRNA restored Cdc42 dependent fluid phase endocytosis in Prom2-expressing cells. Conclusions: Prom2 protrusions primarily

  1. Transanal protrusion of a ventriculoperitoneal shunt catheter.

    PubMed

    Ozturk, Hulya; Is, Merih; Ozturk, Hayrettin; Kucuk, Adem; Dosoglu, Murat

    2012-11-01

    A two years old boy presented with a transanal protrusion of the ventriculoperitoneal (VP) shunt catheter. A VP shunt was inserted when the boy was six-month-old for congenital hydrocephalus. He was active and neurologically normal, with no signs of meningitis and peritoneal irritation. During laparotomy, the tube was seen entering the sigmoid colon, so the tube was cut at the point where it entered the sigmoid colon. The distal end of the protruding tube was pulled out from the anus. The sigmoid colon was repaired, and a delayed shunt revision was completed. The patient was discharged without abdominal and neurological deterioration.

  2. Transanal protrusion of a ventriculoperitoneal shunt catheter.

    PubMed

    Ozturk, Hulya; Is, Merih; Ozturk, Hayrettin; Kucuk, Adem; Dosoglu, Murat

    2012-11-01

    A two years old boy presented with a transanal protrusion of the ventriculoperitoneal (VP) shunt catheter. A VP shunt was inserted when the boy was six-month-old for congenital hydrocephalus. He was active and neurologically normal, with no signs of meningitis and peritoneal irritation. During laparotomy, the tube was seen entering the sigmoid colon, so the tube was cut at the point where it entered the sigmoid colon. The distal end of the protruding tube was pulled out from the anus. The sigmoid colon was repaired, and a delayed shunt revision was completed. The patient was discharged without abdominal and neurological deterioration. PMID:23146859

  3. ERK reinforces actin polymerization to power persistent edge protrusion during motility

    PubMed Central

    Mendoza, Michelle C.; Vilela, Marco; Juarez, Jesus E.; Blenis, John; Danuser, Gaudenz

    2016-01-01

    Cells move through perpetual protrusion and retraction cycles at the leading edge. These cycles are coordinated with substrate adhesion and retraction of the cell rear. Here, we tracked spatial and temporal fluctuations in the molecular activities of individual moving cells to elucidate how extracellular regulated kinase (ERK) signaling controlled the dynamics of protrusion and retraction cycles. ERK is activated by many cell-surface receptors and we found that ERK signaling specifically reinforced cellular protrusions so that they translated into rapid, sustained forward motion of the leading edge. Using quantitative fluorescent speckle microscopy (qFSM) and cross-correlation analysis, we showed that ERK controlled the rate and timing of actin polymerization by promoting the recruitment of the actin nucleator Arp2/3 to the leading edge. Arp2/3 activity generates branched actin networks that can produce pushing force. These findings support a model in which surges in ERK activity induced by extracellular cues enhance Arp2/3-mediated actin polymerization to generate protrusion power phases with enough force to counteract increasing membrane tension and to promote sustained motility. PMID:25990957

  4. FAK phosphorylation at Tyr-925 regulates cross-talk between focal adhesion turnover and cell protrusion

    PubMed Central

    Deramaudt, Therese B.; Dujardin, Denis; Hamadi, Abdelkader; Noulet, Fanny; Kolli, Kaouther; De Mey, Jan; Takeda, Kenneth; Rondé, Philippe

    2011-01-01

     Cell migration is a highly complex process that requires the coordinated formation of membrane protrusion and focal adhesions (FAs). Focal adhesion kinase (FAK), a major signaling component of FAs, is involved in the disassembly process of FAs through phosphorylation and dephosphorylation of its tyrosine residues, but the role of such phosphorylations in nascent FA formation and turnover near the cell front and in cell protrusion is less well understood. In the present study, we demonstrate that, depending on the phosphorylation status of Tyr-925 residue, FAK modulates cell migration via two specific mechanisms. FAK−/− mouse embryonic fibroblasts (MEFs) expressing nonphosphorylatable Y925F-FAK show increased interactions between FAK and unphosphorylated paxillin, which lead to FA stabilization and thus decreased FA turnover and reduced cell migration. Conversely, MEFs expressing phosphomimetic Y925E-FAK display unchanged FA disassembly rates, show increase in phosphorylated paxillin in FAs, and exhibit increased formation of nascent FAs at the cell leading edges. Moreover, Y925E-FAK cells present enhanced cell protrusion together with activation of the p130CAS/Dock180/Rac1 signaling pathway. Together, our results demonstrate that phosphorylation of FAK at Tyr-925 is required for FAK-mediated cell migration and cell protrusion. PMID:21289086

  5. ERK reinforces actin polymerization to power persistent edge protrusion during motility.

    PubMed

    Mendoza, Michelle C; Vilela, Marco; Juarez, Jesus E; Blenis, John; Danuser, Gaudenz

    2015-05-19

    Cells move through perpetual protrusion and retraction cycles at the leading edge. These cycles are coordinated with substrate adhesion and retraction of the cell rear. We tracked spatial and temporal fluctuations in the molecular activities of individual moving cells to elucidate how extracellular signal-regulated kinase (ERK) signaling controlled the dynamics of protrusion and retraction cycles. ERK is activated by many cell surface receptors, and we found that ERK signaling specifically reinforced cellular protrusions so that they translated into rapid, sustained forward motion of the leading edge. Using quantitative fluorescent speckle microscopy and cross-correlation analysis, we showed that ERK controlled the rate and timing of actin polymerization by promoting the recruitment of the actin nucleator Arp2/3 to the leading edge. These findings support a model in which surges in ERK activity induced by extracellular cues enhance Arp2/3-mediated actin polymerization to generate protrusion power phases with enough force to counteract increasing membrane tension and to promote sustained motility.

  6. The class II phosphatidylinositol 3-phosphate kinase PIK3C2A promotes Shigella flexneri dissemination through formation of vacuole-like protrusions.

    PubMed

    Dragoi, Ana-Maria; Agaisse, Hervé

    2015-04-01

    Intracellular pathogens such as Shigella flexneri and Listeria monocytogenes achieve dissemination in the intestinal epithelium by displaying actin-based motility in the cytosol of infected cells. As they reach the cell periphery, motile bacteria form plasma membrane protrusions that resolve into vacuoles in adjacent cells, through a poorly understood mechanism. Here, we report on the role of the class II phosphatidylinositol 3-phosphate kinase PIK3C2A in S. flexneri dissemination. Time-lapse microscopy revealed that PIK3C2A was required for the resolution of protrusions into vacuoles through the formation of an intermediate membrane-bound compartment that we refer to as a vacuole-like protrusion (VLP). Genetic rescue of PIK3C2A depletion with RNA interference (RNAi)-resistant cDNA constructs demonstrated that VLP formation required the activity of PIK3C2A in primary infected cells. PIK3C2A expression was required for production of phosphatidylinositol 3-phosphate [PtdIns(3)P] at the plasma membrane surrounding protrusions. PtdIns(3)P production was not observed in the protrusions formed by L. monocytogenes, whose dissemination did not rely on PIK3C2A. PIK3C2A-mediated PtdIns(3)P production in S. flexneri protrusions was regulated by host cell tyrosine kinase signaling and relied on the integrity of the S. flexneri type 3 secretion system (T3SS). We suggest a model of S. flexneri dissemination in which the formation of VLPs is mediated by the PIK3C2A-dependent production of the signaling lipid PtdIns(3)P in the protrusion membrane, which relies on the T3SS-dependent activation of tyrosine kinase signaling in protrusions.

  7. Regulation of cell protrusions by small GTPases during fusion of the neural folds

    PubMed Central

    Rolo, Ana; Savery, Dawn; Escuin, Sarah; de Castro, Sandra C; Armer, Hannah EJ; Munro, Peter MG; Molè, Matteo A; Greene, Nicholas DE; Copp, Andrew J

    2016-01-01

    Epithelial fusion is a crucial process in embryonic development, and its failure underlies several clinically important birth defects. For example, failure of neural fold fusion during neurulation leads to open neural tube defects including spina bifida. Using mouse embryos, we show that cell protrusions emanating from the apposed neural fold tips, at the interface between the neuroepithelium and the surface ectoderm, are required for completion of neural tube closure. By genetically ablating the cytoskeletal regulators Rac1 or Cdc42 in the dorsal neuroepithelium, or in the surface ectoderm, we show that these protrusions originate from surface ectodermal cells and that Rac1 is necessary for the formation of membrane ruffles which typify late closure stages, whereas Cdc42 is required for the predominance of filopodia in early neurulation. This study provides evidence for the essential role and molecular regulation of membrane protrusions prior to fusion of a key organ primordium in mammalian development. DOI: http://dx.doi.org/10.7554/eLife.13273.001 PMID:27114066

  8. Cortactin Branches Out: Roles in Regulating Protrusive Actin Dynamics

    PubMed Central

    Ammer, Amanda Gatesman; Weed, Scott A.

    2008-01-01

    Since its discovery in the early 1990’s, cortactin has emerged as a key signaling protein in many cellular processes, including cell adhesion, migration, endocytosis, and tumor invasion. While the list of cellular functions influenced by cortactin grows, the ability of cortactin to interact with and alter the cortical actin network is central to its role in regulating these processes. Recently, several advances have been made in our understanding of the interaction between actin and cortactin, providing insight into how these two proteins work together to provide a framework for normal and altered cellular function. This review examines how regulation of cortactin through post-translational modifications and interactions with multiple binding partners elicits changes in cortical actin cytoskeletal organization, impacting the regulation and formation of actin-rich motility structures. PMID:18615630

  9. Nanoscale segregation of actin nucleation and elongation factors determines dendritic spine protrusion

    PubMed Central

    Chazeau, Anaël; Mehidi, Amine; Nair, Deepak; Gautier, Jérémie J; Leduc, Cécile; Chamma, Ingrid; Kage, Frieda; Kechkar, Adel; Thoumine, Olivier; Rottner, Klemens; Choquet, Daniel; Gautreau, Alexis; Sibarita, Jean-Baptiste; Giannone, Grégory

    2014-01-01

    Actin dynamics drive morphological remodeling of neuronal dendritic spines and changes in synaptic transmission. Yet, the spatiotemporal coordination of actin regulators in spines is unknown. Using single protein tracking and super-resolution imaging, we revealed the nanoscale organization and dynamics of branched F-actin regulators in spines. Branched F-actin nucleation occurs at the PSD vicinity, while elongation occurs at the tip of finger-like protrusions. This spatial segregation differs from lamellipodia where both branched F-actin nucleation and elongation occur at protrusion tips. The PSD is a persistent confinement zone for IRSp53 and the WAVE complex, an activator of the Arp2/3 complex. In contrast, filament elongators like VASP and formin-like protein-2 move outwards from the PSD with protrusion tips. Accordingly, Arp2/3 complexes associated with F-actin are immobile and surround the PSD. Arp2/3 and Rac1 GTPase converge to the PSD, respectively, by cytosolic and free-diffusion on the membrane. Enhanced Rac1 activation and Shank3 over-expression, both associated with spine enlargement, induce delocalization of the WAVE complex from the PSD. Thus, the specific localization of branched F-actin regulators in spines might be reorganized during spine morphological remodeling often associated with synaptic plasticity. PMID:25293574

  10. Directional Persistence of EGF-Induced Cell Migration Is Associated with Stabilization of Lamellipodial Protrusions

    PubMed Central

    Harms, Brian D.; Bassi, Gina M.; Horwitz, Alan Rick; Lauffenburger, Douglas A.

    2005-01-01

    Migrating cells can sustain a relatively constant direction of lamellipodial protrusion and locomotion over timescales ranging from minutes to hours. However, individual waves of lamellipodial extension occur over much shorter characteristic times. Little understanding exists regarding how cells might integrate biophysical processes across these disparate timescales to control the directional persistence of locomotion. We address this issue by examining the effects of epidermal growth factor (EGF) stimulation on long-timescale directional persistence and short-timescale lamellipodial dynamics of EGF receptor-transfected Chinese hamster ovary cells migrating on fibronectin-coated substrata. Addition of EGF increased persistence, with the magnitude of increase correlating with fibronectin coating concentration. Kymographic analysis of EGF-stimulated lamellipodial dynamics revealed that the temporal stability of lamellipodial protrusions similarly increased with fibronectin concentration. A soluble RGD peptide competitor reduced both the persistence of long-timescale cell paths and the stability of short-timescale membrane protrusions, indicating that cell-substratum adhesion concomitantly influences lamellipodial dynamics and directional persistence. These results reveal the importance of adhesion strength in regulating the directional motility of cells and suggest that the short-timescale kinetics of adhesion complex formation may play a key role in modulating directional persistence over much longer timescales. PMID:15713602

  11. Differential regulation of protrusion and polarity by PI(3)K during neutrophil motility in live zebrafish

    PubMed Central

    Yoo, Sa Kan; Deng, Qing; Cavnar, Peter J.; Wu, Yi I.; Hahn, Klaus M.; Huttenlocher, Anna

    2010-01-01

    Summary Cell polarity is crucial for directed migration. Here we show that phosphoinositide 3-kinase (PI(3)K) mediates neutrophil migration in vivo by differentially regulating cell protrusion and polarity. The dynamics of PI(3)K products PI(3,4,5)P3-PI(3,4)P2 during neutrophil migration were visualized in living zebrafish, revealing that PI(3)K activation at the leading edge is critical for neutrophil motility in intact tissues. A genetically encoded photoactivatable Rac was used to demonstrate that localized activation of Rac is sufficient to direct migration with precise temporal and spatial control in vivo. Similar stimulation of PI(3)K-inhibited cells did not direct migration. Localized Rac activation rescued membrane protrusion but not anteroposterior polarization of F-actin dynamics of PI(3)K-inhibited cells. Uncoupling Rac-mediated protrusion and polarization suggests a paradigm of two-tiered PI(3)K-mediated regulation of cell motility. This work provides new insight into how cell signaling at the front and back of the cell is coordinated during polarized cell migration in intact tissues within a multicellular organism. PMID:20159593

  12. Influence of bimaxillary protrusion on the perception of smile esthetics

    PubMed Central

    Almutairi, Terki K.; Albarakati, Sahar F.; Aldrees, Abdullah M.

    2015-01-01

    Objectives: To evaluate the impact of bimaxillary protrusion on smile esthetics as perceived by dental professionals and laypersons. Methods: One hundred and fifty evaluators, equally distributed into their respective panels (orthodontists, general dentists, and laypersons), participated in this cross-sectional study conducted in April to December 2012 in Riyadh, Saudi Arabia. The patient sample consisted of 14 female patients divided equally into 2 groups: bimaxillary protrusion patients, and patients who have had 4-premolar extraction treatment. Two standardized photographs (frontal and three-quarter close-up smile views), and a lateral cephalogram were taken for each patient. The evaluators were asked to rate the attractiveness of each photo according to a 100-mm visual analog scale. These esthetic ratings were correlated with the patients’ cephalometric measurements. Results: The bimaxillary protrusion group was rated significantly as less attractive than the treatment group by each evaluator panel. Panel comparison showed that laypeople were less receptive of bimaxillary protrusion than dental professionals. Frontal and three-quarter views of the same smiles were not similarly rated for esthetic perceptions. Correlational analysis revealed that the dentoalveolar measurement with the highest significant negative correlation to the smile esthetics was the upper incisors to palatal plane (U1-PP) angle. Conclusion: Patients with bimaxillary protrusion were found to be less attractive than patients who were treated for the condition. This was especially evident among the laypersons. An increase in the upper incisor inclination, as well as a decrease in the interincisal angle compounds the bimaxillary effect. PMID:25630010

  13. Local force induced conical protrusions of phagocytic cells.

    PubMed

    Vonna, Laurent; Wiedemann, Agnès; Aepfelbacher, Martin; Sackmann, Erich

    2003-03-01

    Magnetic tweezers were used to study the passive and active response of macrophages to local centripetal nanonewton forces on beta1 integrins. Superparamagnetic beads coated with the beta1-integrin-binding protein invasin were attached to J774 murine macrophages to mimic phagocytosis of bacterial pathogens. Forces exceeding approximately 0.5 nN induce the active formation of trumpet-like protrusions resembling pseudopodia after an initial elastic deflection and a response time of approximately 30 seconds. The speed of advancement of the protrusion is =0.065+/-0.020 micro m second(-1) and is force independent. After saturation (after about 100 seconds) the protrusion stops abruptly and is completely retracted again against forces exceeding 5 nN with an effective relaxation time of approximately 30 seconds. The active protrusion is tentatively attributed to the growth of the actin cortex in the direction of the force, and evidence for the involvement of actin is provided by the finding that Latrunculin A abolishes the activated cone growth. The growth is assumed to be activated by cell signaling mediated by the invasin-specific integrins (exhibiting beta1 chains) and could play a role in phagocytic and protrusive events during immune response by macrophages. PMID:12571276

  14. Actin Mediates the Nanoscale Membrane Organization of the Clustered Membrane Protein Influenza Hemagglutinin

    PubMed Central

    Gudheti, Manasa V.; Curthoys, Nikki M.; Gould, Travis J.; Kim, Dahan; Gunewardene, Mudalige S.; Gabor, Kristin A.; Gosse, Julie A.; Kim, Carol H.; Zimmerberg, Joshua; Hess, Samuel T.

    2013-01-01

    The influenza viral membrane protein hemagglutinin (HA) is required at high concentrations on virion and host-cell membranes for infectivity. Because the role of actin in membrane organization is not completely understood, we quantified the relationship between HA and host-cell actin at the nanoscale. Results obtained using superresolution fluorescence photoactivation localization microscopy (FPALM) in nonpolarized cells show that HA clusters colocalize with actin-rich membrane regions (ARMRs). Individual molecular trajectories in live cells indicate restricted HA mobility in ARMRs, and actin disruption caused specific changes to HA clustering. Surprisingly, the actin-binding protein cofilin was excluded from some regions within several hundred nanometers of HA clusters, suggesting that HA clusters or adjacent proteins within the same clusters influence local actin structure. Thus, with the use of imaging, we demonstrate a dynamic relationship between glycoprotein membrane organization and the actin cytoskeleton at the nanoscale. PMID:23708358

  15. Drop stability in wind: effect of solid protrusion

    NASA Astrophysics Data System (ADS)

    Hooshanginejad, Alireza; Wilcox, Benjamin; White, Edward; Lee, Sungyon

    2015-11-01

    We experimentally investigate the inertia-driven onset of droplet depinning behind a solid protrusion inside a wind tunnel. In the high Reynolds number regime, the separation and reattachment of the boundary layer in the presence of the solid protrusion directly lead to complex behavior of the partially wetting water drop as a function of its position. For varying droplet volumes and droplet positions from the protrusion, we measure the critical wind velocity at which the drop starts to depin. In particular, drops in a certain volume range are observed to reverse their depinning direction at a critical distance from the solid. By coupling the boundary layer characteristics with droplet dynamics, we explain the physical mechanism of the resultant droplet behavior.

  16. The Shigella flexneri type 3 secretion system is required for tyrosine kinase-dependent protrusion resolution, and vacuole escape during bacterial dissemination.

    PubMed

    Kuehl, Carole J; Dragoi, Ana-Maria; Agaisse, Hervé

    2014-01-01

    Shigella flexneri is a human pathogen that triggers its own entry into intestinal cells and escapes primary vacuoles to gain access to the cytosolic compartment. As cytosolic and motile bacteria encounter the cell cortex, they spread from cell to cell through formation of membrane protrusions that resolve into secondary vacuoles in adjacent cells. Here, we examined the roles of the Type 3 Secretion System (T3SS) in S. flexneri dissemination in HT-29 intestinal cells infected with the serotype 2a strain 2457T. We generated a 2457T strain defective in the expression of MxiG, a central component of the T3SS needle apparatus. As expected, the ΔmxiG strain was severely affected in its ability to invade HT-29 cells, and expression of mxiG under the control of an arabinose inducible expression system (ΔmxiG/pmxiG) restored full infectivity. In this experimental system, removal of the inducer after the invasion steps (ΔmxiG/pmxiG (Ara withdrawal)) led to normal actin-based motility in the cytosol of HT-29 cells. However, the time spent in protrusions until vacuole formation was significantly increased. Moreover, the number of formed protrusions that failed to resolve into vacuoles was also increased. Accordingly, the ΔmxiG/pmxiG (Ara withdrawal) strain failed to trigger tyrosine phosphorylation in membrane protrusions, a signaling event that is required for the resolution of protrusions into vacuoles. Finally, the ΔmxiG/pmxiG (Ara withdrawal) strain failed to escape from the formed secondary vacuoles, as previously reported in non-intestinal cells. Thus, the T3SS system displays multiple roles in S. flexneri dissemination in intestinal cells, including the tyrosine kinase signaling-dependent resolution of membrane protrusions into secondary vacuoles, and the escape from the formed secondary vacuoles.

  17. Formation of Combined Surface Features of Protrusion Array and Wrinkles atop Shape-Memory Polymer

    NASA Astrophysics Data System (ADS)

    Sun, L.; Zhao, Y.; Huang, W. M.; Tong, T. H.

    We demonstrate a simple and cost-effective approach to realize two combined surface features of different scales together, namely submillimeter-sized protrusion array and microwrinkles, atop a polystyrene shape-memory polymer. Two different types of protrusions, namely flat-top protrusion and crown-shaped protrusion, were studied. The array of protrusions was produced by the Indentation-Polishing-Heating (IPH) process. Compactly packed steel balls were used for making array of indents. A thin gold layer was sputter deposited atop the polymer surface right after polishing. After heating for shape recovery, array of protrusions with wrinkles on the top due to the buckling of gold layer was produced.

  18. Glutamate induces the rapid formation of spine head protrusions in hippocampal slice cultures

    PubMed Central

    Richards, David A.; Mateos, José Maria; Hugel, Sylvain; de Paola, Vincenzo; Caroni, Pico; Gähwiler, Beat H.; McKinney, R. Anne

    2005-01-01

    Synaptic plasticity at neuronal connections has been well characterized functionally by using electrophysiological approaches, but the structural basis for this phenomenon remains controversial. We have studied the dynamic interactions between presynaptic and postsynaptic structures labeled with FM 4-64 and a membrane-targeted GFP, respectively, in hippocampal slices. Under conditions of reduced neuronal activity (1 μM tetrodotoxin), we observed extension of glutamate receptor-dependent processes from dendritic spines of CA1 pyramidal cells to presynaptic boutons. The formation of these spine head protrusions is blocked by α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) receptor antagonists and by agents that reduce the release of glutamate from presynaptic terminals. Moreover, spine head protrusions form in response to exogenously applied glutamate, with clear directionality toward the glutamate electrode. Our results suggest that spontaneously released glutamate is sufficient to activate nearby spines, which can then lead to the growth of new postsynaptic processes connecting to a presynaptic site. Spines thus can compare their recent history with that of neighboring synapses and modify local connectivity accordingly. PMID:15831587

  19. Protrusive and Contractile Forces of Spreading Human Neutrophils

    PubMed Central

    Henry, Steven J.; Chen, Christopher S.; Crocker, John C.; Hammer, Daniel A.

    2015-01-01

    Human neutrophils are mediators of innate immunity and undergo dramatic shape changes at all stages of their functional life cycle. In this work, we quantified the forces associated with a neutrophil’s morphological transition from a nonadherent, quiescent sphere to its adherent and spread state. We did this by tracking, with high spatial and temporal resolution, the cell’s mechanical behavior during spreading on microfabricated post-array detectors printed with the extracellular matrix protein fibronectin. Two dominant mechanical regimes were observed: transient protrusion and steady-state contraction. During spreading, a wave of protrusive force (75 ± 8 pN/post) propagates radially outward from the cell center at a speed of 206 ± 28 nm/s. Once completed, the cells enter a sustained contractile state. Although post engagement during contraction was continuously varying, posts within the core of the contact zone were less contractile (−20 ± 10 pN/post) than those residing at the geometric perimeter (−106 ± 10 pN/post). The magnitude of the protrusive force was found to be unchanged in response to cytoskeletal inhibitors of lamellipodium formation and myosin II-mediated contractility. However, cytochalasin B, known to reduce cortical tension in neutrophils, slowed spreading velocity (61 ± 37 nm/s) without significantly reducing protrusive force. Relaxation of the actin cortical shell was a prerequisite for spreading on post arrays as demonstrated by stiffening in response to jasplakinolide and the abrogation of spreading. ROCK and myosin II inhibition reduced long-term contractility. Function blocking antibody studies revealed haptokinetic spreading was induced by β2 integrin ligation. Neutrophils were found to moderately invaginate the post arrays to a depth of ∼1 μm as measured from spinning disk confocal microscopy. Our work suggests a competition of adhesion energy, cortical tension, and the relaxation of cortical tension is at play at the

  20. Protrusive and Contractile Forces of Spreading Human Neutrophils.

    PubMed

    Henry, Steven J; Chen, Christopher S; Crocker, John C; Hammer, Daniel A

    2015-08-18

    Human neutrophils are mediators of innate immunity and undergo dramatic shape changes at all stages of their functional life cycle. In this work, we quantified the forces associated with a neutrophil's morphological transition from a nonadherent, quiescent sphere to its adherent and spread state. We did this by tracking, with high spatial and temporal resolution, the cell's mechanical behavior during spreading on microfabricated post-array detectors printed with the extracellular matrix protein fibronectin. Two dominant mechanical regimes were observed: transient protrusion and steady-state contraction. During spreading, a wave of protrusive force (75 ± 8 pN/post) propagates radially outward from the cell center at a speed of 206 ± 28 nm/s. Once completed, the cells enter a sustained contractile state. Although post engagement during contraction was continuously varying, posts within the core of the contact zone were less contractile (-20 ± 10 pN/post) than those residing at the geometric perimeter (-106 ± 10 pN/post). The magnitude of the protrusive force was found to be unchanged in response to cytoskeletal inhibitors of lamellipodium formation and myosin II-mediated contractility. However, cytochalasin B, known to reduce cortical tension in neutrophils, slowed spreading velocity (61 ± 37 nm/s) without significantly reducing protrusive force. Relaxation of the actin cortical shell was a prerequisite for spreading on post arrays as demonstrated by stiffening in response to jasplakinolide and the abrogation of spreading. ROCK and myosin II inhibition reduced long-term contractility. Function blocking antibody studies revealed haptokinetic spreading was induced by β2 integrin ligation. Neutrophils were found to moderately invaginate the post arrays to a depth of ∼1 μm as measured from spinning disk confocal microscopy. Our work suggests a competition of adhesion energy, cortical tension, and the relaxation of cortical tension is at play at the onset of

  1. Protrusive and Contractile Forces of Spreading Human Neutrophils.

    PubMed

    Henry, Steven J; Chen, Christopher S; Crocker, John C; Hammer, Daniel A

    2015-08-18

    Human neutrophils are mediators of innate immunity and undergo dramatic shape changes at all stages of their functional life cycle. In this work, we quantified the forces associated with a neutrophil's morphological transition from a nonadherent, quiescent sphere to its adherent and spread state. We did this by tracking, with high spatial and temporal resolution, the cell's mechanical behavior during spreading on microfabricated post-array detectors printed with the extracellular matrix protein fibronectin. Two dominant mechanical regimes were observed: transient protrusion and steady-state contraction. During spreading, a wave of protrusive force (75 ± 8 pN/post) propagates radially outward from the cell center at a speed of 206 ± 28 nm/s. Once completed, the cells enter a sustained contractile state. Although post engagement during contraction was continuously varying, posts within the core of the contact zone were less contractile (-20 ± 10 pN/post) than those residing at the geometric perimeter (-106 ± 10 pN/post). The magnitude of the protrusive force was found to be unchanged in response to cytoskeletal inhibitors of lamellipodium formation and myosin II-mediated contractility. However, cytochalasin B, known to reduce cortical tension in neutrophils, slowed spreading velocity (61 ± 37 nm/s) without significantly reducing protrusive force. Relaxation of the actin cortical shell was a prerequisite for spreading on post arrays as demonstrated by stiffening in response to jasplakinolide and the abrogation of spreading. ROCK and myosin II inhibition reduced long-term contractility. Function blocking antibody studies revealed haptokinetic spreading was induced by β2 integrin ligation. Neutrophils were found to moderately invaginate the post arrays to a depth of ∼1 μm as measured from spinning disk confocal microscopy. Our work suggests a competition of adhesion energy, cortical tension, and the relaxation of cortical tension is at play at the onset of

  2. Membrane tubulation from giant lipid vesicles in alternating electric fields.

    PubMed

    Antonova, K; Vitkova, V; Meyer, C

    2016-01-01

    We report on the formation of tubular membrane protrusions from giant unilamellar vesicles in alternating electric fields. The construction of the experimental chamber permitted the application of external AC fields with strength of dozens of V/mm and kHz frequency during relatively long time periods (several minutes). Besides the vesicle electrodeformation from quasispherical to prolate ellipsoidal shape, the formation of long tubular membrane protrusions with length of up to several vesicle diameters, arising from the vesicular surface in the field direction, was registered and analyzed. The threshold electric field at which the electro-induced protrusions appeared was lower than the field strengths inducing membrane electroporation. PMID:26871107

  3. Shape control of lipid bilayer membranes by confined actin bundles.

    PubMed

    Tsai, Feng-Ching; Koenderink, Gijsje Hendrika

    2015-12-01

    In living cells, lipid membranes and biopolymers determine each other's conformation in a delicate force balance. Cellular polymers such as actin filaments are strongly confined by the plasma membrane in cell protrusions such as lamellipodia and filopodia. Conversely, protrusion formation is facilitated by actin-driven membrane deformation and these protrusions are maintained by dense actin networks or bundles of actin filaments. Here we investigate the mechanical interplay between actin bundles and lipid bilayer membranes by reconstituting a minimal model system based on cell-sized liposomes with encapsulated actin filaments bundled by fascin. To address the competition between the deformability of the membrane and the enclosed actin bundles, we tune the bundle stiffness (through the fascin-to-actin molar ratio) and the membrane rigidity (through protein decoration). Using confocal microscopy and quantitative image analysis, we show that actin bundles deform the liposomes into a rich set of morphologies. For liposomes having a small membrane bending rigidity, the actin bundles tend to generate finger-like membrane protrusions that resemble cellular filopodia. Stiffer bundles formed at high crosslink density stay straight in the liposome body, whereas softer bundles formed at low crosslink density are bent and kinked. When the membrane has a large bending rigidity, membrane protrusions are suppressed. In this case, membrane enclosure forces the actin bundles to organize into cortical rings, to minimize the energy cost associated with filament bending. Our results highlight the importance of taking into account mechanical interactions between the actin cytoskeleton and the membrane to understand cell shape control.

  4. PTEN knockdown alters dendritic spine/protrusion morphology, not density

    PubMed Central

    Haws, Michael E.; Jaramillo, Thomas C.; Espinosa-Becerra, Felipe; Widman, Allie; Stuber, Garret D.; Sparta, Dennis R.; Tye, Kay M.; Russo, Scott J.; Parada, Luis F.; Kaplitt, Michael; Bonci, Antonello; Powell, Craig M.

    2014-01-01

    Mutations in phosphatase and tensin homolog deleted on chromosome ten (PTEN) are implicated in neuropsychiatric disorders including autism. Previous studies report that PTEN knockdown in neurons in vivo leads to increased spine density and synaptic activity. To better characterize synaptic changes in neurons lacking PTEN, we examined the effects of shRNA knockdown of PTEN in basolateral amygdala neurons on synaptic spine density and morphology using fluorescent dye confocal imaging. Contrary to previous studies in dentate gyrus, we find that knockdown of PTEN in basolateral amygdala leads to a significant decrease in total spine density in distal dendrites. Curiously, this decreased spine density is associated with increased miniature excitatory post-synaptic current frequency and amplitude, suggesting an increase in number and function of mature spines. These seemingly contradictory findings were reconciled by spine morphology analysis demonstrating increased mushroom spine density and size with correspondingly decreased thin protrusion density at more distal segments. The same analysis of PTEN conditional deletion in dentate gyrus demonstrated that loss of PTEN does not significantly alter total density of dendritic protrusions in the dentate gyrus, but does decrease thin protrusion density and increases density of more mature mushroom spines. These findings suggest that, contrary to previous reports, PTEN knockdown may not induce de novo spinogenesis, but instead may increase synaptic activity by inducing morphological and functional maturation of spines. Furthermore, behavioral analysis of basolateral amygdala PTEN knockdown suggests that these changes limited only to the basolateral amygdala complex may not be sufficient to induce increased anxiety-related behaviors. PMID:24264880

  5. Management of severe pain due to lumbar disk protrusion.

    PubMed

    Conroy, Liam

    2015-03-01

    Lumbar intervertebral disk protrusion can cause excruciating pain in severe cases, which can be exacerbated by activity such as sitting down and straining at stool. Acute sciatica due to disk rupture will improve within 1 to 3 months. The efficacy of drugs used for the management of sciatica in primary care is unclear. Severe cases can require opioid analgesia, however people taking opioids for pain relief frequently present with opioid-induced bowel dysfunction. The use of transforaminal steroid injections is a controversial issue and repeat steroid injections should be considered in light of the risk-benefit profile of the individual patient. PMID:25643230

  6. Counterion density profiles at charged flexible membranes.

    PubMed

    Fleck, Christian C; Netz, Roland R

    2005-09-16

    Counterion distributions at charged flexible membranes are studied using analytical and simulation methods in both Poisson-Boltzmann and strong-coupling limits. The softer the membrane, the more smeared out the counterion-density profile becomes and counterions penetrate through the mean-membrane surface location, in agreement with anomalous scattering results. Repulsion between membrane charges enhances protrusions and induces short-scale membrane roughening.

  7. [Two Cases of Ruptured Cerebral Aneurysm Complicated with Delayed Coil Protrusion after Coil Embolization].

    PubMed

    Furukawa, Takashi; Ogata, Atsushi; Ebashi, Ryo; Takase, Yukinori; Masuoka, Jun; Kawashima, Masatou; Abe, Tatsuya

    2016-07-01

    We report two cases of delayed coil protrusion after coil embolization for ruptured cerebral aneurysms. Case 1:An 82-year-old woman with a subarachnoid hemorrhage due to a ruptured small anterior communicating artery aneurysm underwent successful coil embolization. Eighteen days after the procedure, coil protrusion from the aneurysm into the right anterior cerebral artery was observed without any symptoms. Further coil protrusion did not develop after 28 days. Case 2:A 78-year-old woman with a subarachnoid hemorrhage due to a ruptured small left middle cerebral artery aneurysm underwent successful coil embolization. Twenty days after the procedure, coil protrusion from the aneurysm into the left middle cerebral artery was observed, with a transient ischemic attack. Further coil protrusion did not develop. Both patients recovered with antithrombotic treatment. Even though delayed coil protrusion after coil embolization is rare, it should be recognized as a long-term complication of coil embolization for cerebral aneurysms. PMID:27384117

  8. Asymmetric formation of coated pits on dorsal and ventral surfaces at the leading edges of motile cells and on protrusions of immobile cells

    PubMed Central

    Kural, Comert; Akatay, Ahmet Ata; Gaudin, Raphaël; Chen, Bi-Chang; Legant, Wesley R.; Betzig, Eric; Kirchhausen, Tom

    2015-01-01

    Clathrin/AP2-coated vesicles are the principal endocytic carriers originating at the plasma membrane. In the experiments reported here, we used spinning-disk confocal and lattice light-sheet microscopy to study the assembly dynamics of coated pits on the dorsal and ventral membranes of migrating U373 glioblastoma cells stably expressing AP2 tagged with enhanced green fluorescence (AP2-EGFP) and on lateral protrusions from immobile SUM159 breast carcinoma cells, gene-edited to express AP2-EGFP. On U373 cells, coated pits initiated on the dorsal membrane at the front of the lamellipodium and at the approximate boundary between the lamellipodium and lamella and continued to grow as they were swept back toward the cell body; coated pits were absent from the corresponding ventral membrane. We observed a similar dorsal/ventral asymmetry on membrane protrusions from SUM159 cells. Stationary coated pits formed and budded on the remainder of the dorsal and ventral surfaces of both types of cells. These observations support a previously proposed model that invokes net membrane deposition at the leading edge due to an imbalance between the endocytic and exocytic membrane flow at the front of a migrating cell. PMID:25851602

  9. Retreatment of a transfer patient with bialveolar protrusion with mini bone-plate anchorage.

    PubMed

    Bilodeau, John E

    2014-10-01

    A 40-year-old black woman, with pretorqued, preangulated appliances on the maxillary teeth, transferred with chief concerns of missing teeth and her orthodontic progress. She had bialveolar protrusion and edentulous areas in both the maxilla and the mandible. After records were made and her desires assessed, she was retreated with the extraction of 3 premolars. Her protrusion was reduced and all edentulous spaces were closed. The facial change was evident due to the reduction of the protrusion. PMID:25263153

  10. Excitable actin dynamics in lamellipodial protrusion and retraction.

    PubMed

    Ryan, Gillian L; Petroccia, Heather M; Watanabe, Naoki; Vavylonis, Dimitrios

    2012-04-01

    Many animal cells initiate crawling by protruding lamellipodia, consisting of a dense network of actin filaments, at their leading edge. We imaged XTC cells that exhibit flat lamellipodia on poly-L-lysine-coated coverslips. Using active contours, we tracked the leading edge and measured the total amount of F-actin by summing the pixel intensities within a 5-μm band. We observed protrusion and retraction with period 130-200 s and local wavelike features. Positive (negative) velocities correlated with minimum (maximum) integrated actin concentration. Approximately constant retrograde flow indicated that protrusions and retractions were driven by fluctuations of the actin polymerization rate. We present a model of these actin dynamics as an excitable system in which a diffusive, autocatalytic activator causes actin polymerization; F-actin accumulation in turn inhibits further activator accumulation. Simulations of the model reproduced the pattern of actin polymerization seen in experiments. To explore the model's assumption of an autocatalytic activation mechanism, we imaged cells expressing markers for both F-actin and the p21 subunit of the Arp2/3 complex. We found that integrated Arp2/3-complex concentrations spike several seconds before spikes of F-actin concentration. This suggests that the Arp2/3 complex participates in an activation mechanism that includes additional diffuse components. Response of cells to stimulation by fetal calf serum could be reproduced by the model, further supporting the proposed dynamical picture.

  11. Tongue Protrusion Strength in Arousal State Is Predictive of the Airway Patency in Obstructive Sleep Apnea.

    PubMed

    Kanezaki, Masashi; Ogawa, Teruhiro; Izumi, Tadafumi

    2015-01-01

    Contraction of the genioglossus affects either tongue protrusion strength or dilating forces of the upper airway. The upper airway in patients with obstructive sleep apnea (OSA) is thought to collapse during sleep, at least in part because of a sleep related reduction in genioglossus muscle activity. Thus, although tongue protrusion strength by genioglossus activity during sleep contributes to the maintenance of airway patency in patients with OSA, the relationship between tongue protrusion strength in the arousal state and obstructive sleep apnea has not been fully elucidated. Conventional method of tongue protrusion strength cannot be used to evaluate in edentulous subjects and/or subjects with the decreased biting force. In this study, employing a novel measurement method that does not require biting a transducer, we investigated relationships between the tongue protrusion strength and polysomnographic findings. We enrolled twenty normal subjects and 26 subjects with OSA. All subjects completed the measurement of tongue protrusion strength. Each subject with OSA was evaluated by full polysomnography. The degree of tongue protrusion strength was assessed by maximum voluntary contraction against the tongue depressor connected with a strain gauge dynamometer. The tongue protrusion strength was negatively correlated with obstructive apnea time, apnea index (AI) and the percent of sleep stage 2 (r = -0.61, p < 0.0001, r = -0.41 p = 0.03 and r = -0.39 p = 0.04, respectively). Tongue protrusion strength measured in the arousal state is predictive of the airway patency during sleep in OSA.

  12. The Rise of Jaw Protrusion in Spiny-Rayed Fishes Closes the Gap on Elusive Prey.

    PubMed

    Bellwood, David R; Goatley, Christopher H R; Bellwood, Orpha; Delbarre, Daniel J; Friedman, Matt

    2015-10-19

    Jaw protrusion is one of the most important innovations in vertebrate feeding over the last 400 million years [1, 2]. Protrusion enables a fish to rapidly decrease the distance between itself and its prey [2, 3]. We assessed the evolution and functional implications of jaw protrusion in teleost fish assemblages from shallow coastal seas since the Cretaceous. By examining extant teleost fishes, we identified a robust morphological predictor of jaw protrusion that enabled us to predict the extent of jaw protrusion in fossil fishes. Our analyses revealed increases in both average and maximum jaw protrusion over the last 100 million years, with a progressive increase in the potential impact of fish predation on elusive prey. Over this period, the increase in jaw protrusion was initially driven by a taxonomic restructuring of fish assemblages, with an increase in the proportion of spiny-rayed fishes (Acanthomorpha), followed by an increase in the extent of protrusion within this clade. By increasing the ability of fishes to catch elusive prey [2, 4], jaw protrusion is likely to have fundamentally changed the nature of predator-prey interactions and may have contributed to the success of the spiny-rayed fishes, the dominant fish clade in modern oceans [5]. PMID:26455299

  13. [The role of intravertebral disk protrusion in pathogenesis of lumbosacral radiculopathies in flying personnel].

    PubMed

    Vit'ko, N K

    1999-01-01

    The significance of x-ray computer tomography in determining the mechanisms of pathogenesis of lumbar-sacral radiculopathy in patients with protrusions of intervertebral disks of the lumbar-sacral backbone is demonstrated. Pilots with the protrusions (n = 135) and the intervertebral disk herinas (79) have been examined. The clinical course of the sickness was reviewed in both groups of patients. The computer-tomographic semiotics of various protrusions was elaborated and expanded and their clinical implications demonstrated. The fresh paradigm of pathogenesis of radiculopathy at various forms of protrusions is presented. The roentgenomorphologic classification of arthrosis of archiprossesed joints was studied and proposed.

  14. The Rise of Jaw Protrusion in Spiny-Rayed Fishes Closes the Gap on Elusive Prey.

    PubMed

    Bellwood, David R; Goatley, Christopher H R; Bellwood, Orpha; Delbarre, Daniel J; Friedman, Matt

    2015-10-19

    Jaw protrusion is one of the most important innovations in vertebrate feeding over the last 400 million years [1, 2]. Protrusion enables a fish to rapidly decrease the distance between itself and its prey [2, 3]. We assessed the evolution and functional implications of jaw protrusion in teleost fish assemblages from shallow coastal seas since the Cretaceous. By examining extant teleost fishes, we identified a robust morphological predictor of jaw protrusion that enabled us to predict the extent of jaw protrusion in fossil fishes. Our analyses revealed increases in both average and maximum jaw protrusion over the last 100 million years, with a progressive increase in the potential impact of fish predation on elusive prey. Over this period, the increase in jaw protrusion was initially driven by a taxonomic restructuring of fish assemblages, with an increase in the proportion of spiny-rayed fishes (Acanthomorpha), followed by an increase in the extent of protrusion within this clade. By increasing the ability of fishes to catch elusive prey [2, 4], jaw protrusion is likely to have fundamentally changed the nature of predator-prey interactions and may have contributed to the success of the spiny-rayed fishes, the dominant fish clade in modern oceans [5].

  15. A new pathological classification of lumbar disc protrusion and its clinical significance.

    PubMed

    Ma, Xin-long

    2015-02-01

    Lumbar disc protrusion is common. Its clinical manifestations and treatments are closely related to the pathological changes; however, the pathological classification of lumbar disc protrusion is controversial. This article introduces a new pathological classification comprising four types of lumbar disc protrusion according to intraoperative findings. The damage-herniation type is probably caused by injury and is characterized by soft herniation, the capsule can easily be cut and the broken disc tissue blocks overflow or is easily removed. The broken disc substances should be completely removed; satisfactory results can be achieved by minimally invasive endoscopic surgery. The degeneration-protrusion type is characterized by hard and tough protrusions and the pathological process by degeneration and proliferative reaction. The nerve should be decompressed and relaxed with minimally invasive removal of the posterior wall; the bulged or protruded disc often need not be excised. The posterior vertebral osteochondrosis with disc protrusion type is characterized by deformity of the posterior vertebral body, osteochondral nodules and intervertebral disc protrusion. The herniated and fragmented disc tissue should be removed with partially protruding osteochondral nodules. Intervertebral disc cyst is of uncertain pathogenesis and is characterized by a cyst that communicates with the disc. Resection of the cyst under microscopic or endoscopic control can achieve good results; and whether the affected disc needs to be simultaneously resected is controversial. The new pathological classification proposed here is will aid better understanding of pathological changes and pathogenesis of lumbar disc protrusion and provides a reference for diagnosis and treatment. PMID:25708029

  16. Real-time monitoring of cell protrusion dynamics by impedance responses

    PubMed Central

    Armando Gagliardi, Paolo; Puliafito, Alberto; di Blasio, Laura; Chianale, Federica; Somale, Desiana; Seano, Giorgio; Bussolino, Federico; Primo, Luca

    2015-01-01

    Cellular protrusions are highly dynamic structures involved in fundamental processes, including cell migration and invasion. For a cell to migrate, its leading edge must form protrusions, and then adhere or retract. The spatial and temporal coordination of protrusions and retraction is yet to be fully understood. The study of protrusion dynamics mainly relies on live-microscopy often coupled to fluorescent labeling. Here we report the use of an alternative, label-free, quantitative and rapid assay to analyze protrusion dynamics in a cell population based on the real-time recording of cell activity by means of electronic sensors. Cells are seeded on a plate covered with electrodes and their shape changes map into measured impedance variations. Upon growth factor stimulation the impedance increases due to protrusive activity and decreases following retraction. Compared to microscopy-based methods, impedance measurements are suitable to high-throughput studies on different cell lines, growth factors and chemical compounds. We present data indicating that this assay lends itself to dissect the biochemical signaling pathways controlling adhesive protrusions. Indeed, we show that the protrusion phase is sustained by actin polymerization, directly driven by growth factor stimulation. Contraction instead mainly relies on myosin action, pointing at a pivotal role of myosin in lamellipodia retraction. PMID:25976978

  17. Carbon protrusions on PTFE surface prepared by ion irradiation and chemical defluorination

    NASA Astrophysics Data System (ADS)

    Kobayashi, T.; Iwaki, M.

    2006-01-01

    A surface of PTFE was covered with small protrusions by ion-beam irradiation. In this study, we converted PTFE protrusions into carbon protrusions by a defluorination (carbonization) process using sodium vapor. The morphology, composition and structure were analyzed by SEM-EDX, Raman spectroscopy and TEM. The irradiated PTFE sheets were packed in evacuated glass tubes with a sodium block and kept at 473 K for 2-48 h. The samples were then rinsed in HCl and distilled water to remove NaF precipitates. The EDX measurement showed that the NaF precipitates were completely removed by washing, and the percentage of carbon atoms was controlled from 60% to 99% by the treatment. Raman spectra showed that graphite structures grow during the defluorination process. TEM micrographs showed that the protrusions have a bubble structure and are covered with a thin wall. The carbonized protrusions were conductive and grew perpendicular to the substrate.

  18. Refilins are short-lived Actin-bundling proteins that regulate lamellipodium protrusion dynamics

    PubMed Central

    Gay, Olivia; Gilquin, Benoît; Assard, Nicole; Stuelsatz, Pascal; Delphin, Christian; Lachuer, Joël; Gidrol, Xavier; Baudier, Jacques

    2016-01-01

    ABSTRACT Refilins (RefilinA and RefilinB) are members of a novel family of Filamin binding proteins that function as molecular switches to conformationally alter the Actin filament network into bundles. We show here that Refilins are extremely labile proteins. An N-terminal PEST/DSG(X)2-4S motif mediates ubiquitin-independent rapid degradation. A second degradation signal is localized within the C-terminus. Only RefilinB is protected from rapid degradation by an auto-inhibitory domain that masks the PEST/DSG(X)2-4S motif. Dual regulation of RefilinA and RefilinB stability was confirmed in rat brain NG2 precursor cells (polydendrocyte). Using loss- and gain-of-function approaches we show that in these cells, and in U373MG cells, Refilins contribute to the dynamics of lamellipodium protrusion by catalysing Actin bundle formation within the lamella Actin network. These studies extend the Actin bundling function of the Refilin-Filamin complex to dynamic regulation of cell membrane remodelling. PMID:27744291

  19. Independently evolved upper jaw protrusion mechanisms show convergent hydrodynamic function in teleost fishes.

    PubMed

    Staab, Katie Lynn; Holzman, Roi; Hernandez, L Patricia; Wainwright, Peter C

    2012-05-01

    A protrusible upper jaw has independently evolved multiple times within teleosts and has been implicated in the success of two groups in particular: Acanthomorpha and Cypriniformes. We use digital particle image velocimetry (DPIV) to compare suction feeding flow dynamics in a representative of each of these clades: goldfish and bluegill. Using DPIV, we contrast the spatial pattern of flow, the temporal relationship between flow and head kinematics, and the contribution of jaw protrusion to the forces exerted on prey. As expected, the spatial patterns of flow were similar in the two species. However, goldfish were slower to reach maximal kinematic excursions, and were more flexible in the relative timing of jaw protrusion, other jaw movements and suction flows. Goldfish were also able to sustain flow speeds for a prolonged period of time as compared with bluegill, in part because goldfish generate lower peak flow speeds. In both species, jaw protrusion increased the force exerted on the prey. However, slower jaw protrusion in goldfish resulted in less augmentation of suction forces. This difference in force exerted on prey corresponds with differences in trophic niches and feeding behavior of the two species. The bluegill uses powerful suction to capture insect larvae whereas the goldfish uses winnowing to sort through detritus and sediment. The kinethmoid of goldfish may permit jaw protrusion that is independent of lower jaw movement, which could explain the ability of goldfish to decouple suction flows (due to buccal expansion) from upper jaw protrusion. Nevertheless, our results show that jaw protrusion allows both species to augment the force exerted on prey, suggesting that this is a fundamental benefit of jaw protrusion to suction feeders. PMID:22496281

  20. Morphological change of self-organized protrusions of fluoropolymer surface by ion beam irradiation

    NASA Astrophysics Data System (ADS)

    Kitamura (Ogawa), Akane; Kobayashi, Tomohiro; Satoh, Takahiro; Koka, Masashi; Kamiya, Tomihiro; Suzuki, Akihiro; Terai, Takayuki

    2013-07-01

    Polytetrafluoroethylene (PTFE) and fluorinated ethylene propylene (FEP) are typical fluoropolymers displaying several desirable technological properties such as electrical insulation and high chemical resistance. When their surfaces are irradiated with ion beams, dense micro-protrusions formed after the emergence and spread of micropores across the entire irradiated area, allowing culture cells to spread on the top of the protrusions. In this study, we investigate the morphological changes introduced in the fluoropolymer surfaces by ion beams as the energy of the beams is increased. When an FEP sample was irradiated with a nitrogen ion beam with an energy of less than 350 keV at 1.0 μA/cm2, protrusions were formed with a density between 2 × 107/cm2 and 2 × 108/cm2. However, at energies higher than 350 keV, the protrusions became sparse, and the density dropped to 5 × 102/cm2. Protrusions appeared sporadically during irradiation at high energies, and the top of the protrusions appeared as spots inside the sample, which were difficult to etch and became elongated as the erosion of the surface progressed. Erosion was caused by sputtering of FEP molecules and evaporation at notably elevated temperatures on the surface. Analysis based on attenuated total reflectance/Fourier transform infrared spectroscopy showed the presence of Cdbnd C bonds as well as -COOH, -Cdbnd O, and -OH bonds on all irradiated samples. Their concentration on the surface densely covered with micro-protrusions was higher than that on the surface with sparse protrusions after irradiation at energies exceeding 350 keV. Thus, we determined a suitable range for the ion energy for creating FEP surfaces densely covered with protrusions.

  1. PAOO technique for the bimaxillary protrusion: Perio-ortho interrelationship

    PubMed Central

    Bhat, Subraya G.; Singh, Vishal; Bhat, Mahalinga K.

    2012-01-01

    An increasing number of adult patients have been seeking orthodontic treatment, and a short treatment time has been a recurring request. To meet their expectations, a number of innovative techniques have been developed to accelerate orthodontic tooth movement. Significant acceleration in orthodontic tooth movement has been extensively reported following a combination of selective alveolar decortication and bone grafting surgery with the latter being responsible for the increased scope of tooth movement and the long-term improvement of the periodontium. Six patients who have been diagnosed as class I malocclusion with bimaxillary protrusion were selected. A modified corticotomy procedure was carried out The active orthodontic treatment was begun within 1 week after surgery and follow up. All the patients reported the minmalpain and mild swelling 2 days following the surgical procedure. The mean treatment time for these patients was 17.4 months, with the distalization of the canines being mostly completed in 8.5 months. Periodontallyaccelerated osteogenicsorthodontic tooth movement procedure or selective alveolar decortications or corticotomy approach is one of the surgical techniques developed to reduce the time for orthodontic treatment. This newer approach is leading to short orthodontic treatment time and great patient acceptance. PMID:23493808

  2. Protrusive waves guide 3D cell migration along nanofibers.

    PubMed

    Guetta-Terrier, Charlotte; Monzo, Pascale; Zhu, Jie; Long, Hongyan; Venkatraman, Lakshmi; Zhou, Yue; Wang, PeiPei; Chew, Sing Yian; Mogilner, Alexander; Ladoux, Benoit; Gauthier, Nils C

    2015-11-01

    In vivo, cells migrate on complex three-dimensional (3D) fibrous matrices, which has made investigation of the key molecular and physical mechanisms that drive cell migration difficult. Using reductionist approaches based on 3D electrospun fibers, we report for various cell types that single-cell migration along fibronectin-coated nanofibers is associated with lateral actin-based waves. These cyclical waves have a fin-like shape and propagate up to several hundred micrometers from the cell body, extending the leading edge and promoting highly persistent directional movement. Cells generate these waves through balanced activation of the Rac1/N-WASP/Arp2/3 and Rho/formins pathways. The waves originate from one major adhesion site at leading end of the cell body, which is linked through actomyosin contractility to another site at the back of the cell, allowing force generation, matrix deformation and cell translocation. By combining experimental and modeling data, we demonstrate that cell migration in a fibrous environment requires the formation and propagation of dynamic, actin based fin-like protrusions.

  3. Protrusive waves guide 3D cell migration along nanofibers

    PubMed Central

    Guetta-Terrier, Charlotte; Monzo, Pascale; Zhu, Jie; Long, Hongyan; Venkatraman, Lakshmi; Zhou, Yue; Wang, PeiPei; Chew, Sing Yian; Mogilner, Alexander

    2015-01-01

    In vivo, cells migrate on complex three-dimensional (3D) fibrous matrices, which has made investigation of the key molecular and physical mechanisms that drive cell migration difficult. Using reductionist approaches based on 3D electrospun fibers, we report for various cell types that single-cell migration along fibronectin-coated nanofibers is associated with lateral actin-based waves. These cyclical waves have a fin-like shape and propagate up to several hundred micrometers from the cell body, extending the leading edge and promoting highly persistent directional movement. Cells generate these waves through balanced activation of the Rac1/N-WASP/Arp2/3 and Rho/formins pathways. The waves originate from one major adhesion site at leading end of the cell body, which is linked through actomyosin contractility to another site at the back of the cell, allowing force generation, matrix deformation and cell translocation. By combining experimental and modeling data, we demonstrate that cell migration in a fibrous environment requires the formation and propagation of dynamic, actin based fin-like protrusions. PMID:26553933

  4. Increased cellular apoptosis susceptibility (CSE1L/CAS) protein expression promotes protrusion extension and enhances migration of MCF-7 breast cancer cells

    SciTech Connect

    Tai, Cheng-Jeng; Shen, Shing-Chuan; Lee, Woan-Ruoh; Liao, Ching-Fong; Deng, Win-Ping; Chiou, Hung-Yi; Hsieh, Cheng-I; Tung, Jai-Nien; Chen, Ching-Shyang; Chiou, Jeng-Fong; Li, Li-Tzu; Lin, Chuang-Yu; Hsu, Chung-Huei; Jiang, Ming-Chung

    2010-10-15

    Microtubules are part of cell structures that play a role in regulating the migration of cancer cells. The cellular apoptosis susceptibility (CSE1L/CAS) protein is a microtubule-associated protein that is highly expressed in cancer. We report here that CSE1L regulates the association of {alpha}-tubulin with {beta}-tubulin and promotes the migration of MCF-7 breast cancer cells. CSE1L was associated with {alpha}-tubulin and {beta}-tubulin in GST (glutathione S-transferase) pull-down and immunoprecipitation assays. CSE1L-GFP (green fluorescence protein) fusion protein experiments showed that the N-terminal of CSE1L interacted with microtubules. Increased CSE1L expression resulted in decreased tyrosine phosphorylation of {alpha}-tubulin and {beta}-tubulin, increased {alpha}-tubulin and {beta}-tubulin association, and enhanced assembly of microtubules. Cell protrusions or pseudopodia are temporary extensions of the plasma membrane and are implicated in cancer cell migration and invasion. Increased CSE1L expression increased the extension of MCF-7 cell protrusions. In vitro migration assay showed that enhanced CSE1L expression increased the migration of MCF-7 cells. Our results indicate that CSE1L plays a role in regulating the extension of cell protrusions and promotes the migration of cancer cells.

  5. Spatio-temporal co-ordination of RhoA, Rac1 and Cdc42 activation during prototypical edge protrusion and retraction dynamics

    PubMed Central

    Martin, Katrin; Reimann, Andreas; Fritz, Rafael D.; Ryu, Hyunryul; Jeon, Noo Li; Pertz, Olivier

    2016-01-01

    The three canonical Rho GTPases RhoA, Rac1 and Cdc42 co-ordinate cytoskeletal dynamics. Recent studies indicate that all three Rho GTPases are activated at the leading edge of motile fibroblasts, where their activity fluctuates at subminute time and micrometer length scales. Here, we use a microfluidic chip to acutely manipulate fibroblast edge dynamics by applying pulses of platelet-derived growth factor (PDGF) or the Rho kinase inhibitor Y-27632 (which lowers contractility). This induces acute and robust membrane protrusion and retraction events, that exhibit stereotyped cytoskeletal dynamics, allowing us to fairly compare specific morphodynamic states across experiments. Using a novel Cdc42, as well as previously described, second generation RhoA and Rac1 biosensors, we observe distinct spatio-temporal signaling programs that involve all three Rho GTPases, during protrusion/retraction edge dynamics. Our results suggest that Rac1, Cdc42 and RhoA regulate different cytoskeletal and adhesion processes to fine tune the highly plastic edge protrusion/retraction dynamics that power cell motility. PMID:26912264

  6. F-actin crosslinker: A key player for the mechanical stability of filopodial protrusion

    NASA Astrophysics Data System (ADS)

    Li, Tong; Oloyede, Adekunle; Gu, Y. T.

    2013-12-01

    Filopodial protrusion initiates cell migration, which decides the fate of cells in biological environments. In order to understand the structural stability of ultra-slender filopodial protrusion, we have developed an explicit modeling strategy that can study both static and dynamic characteristics of microfilament bundles. Our study reveals that the stability of filopodial protrusions is dependent on the density of F-actin crosslinkers. This cross-linkage strategy is a requirement for the optimization of cell structures, resulting in the provision and maintenance of adequate bending stiffness and buckling resistance while mediating the vibration. This cross-linkage strategy explains the mechanical stability of filopodial protrusion and helps understand the mechanisms of mechanically induced cellular activities.

  7. Evaluation and Management of Plaque Protrusion or Thrombus following Carotid Artery Stenting

    PubMed Central

    HASHIMURA, Naoki; MUTOH, Tatsushi; MATSUDA, Kazuya; MATSUMOTO, Keigo

    2015-01-01

    Carotid artery stenting (CAS) has become a common treatment for carotid artery stenosis. However, complications, such as an ischemic event, can occur with CAS during intra- and post-operative periods. Among these ischemic complications, plaque protrusion into the stent and thrombus on the stent have occurred after CAS. We retrospectively evaluated the temporal profile and treatment options for these complications in 32 consecutive cases who underwent CAS at our hospital between April 2009 and December 2011. The cases were evaluated pre-operatively for risk factors, as well as the plaque morphology and characteristics using computed tomographic angiography (CTA), ultrasound (US), and magnetic resonance imaging (MRI). Post-operatively, lesions were examined by CTA and/or US within 1 week of CAS. As a result, among the 32 cases, 8 experienced plaque protrusions or thrombus, which were treated with medication (anti-platelet and/or anti-coagulation reinforcement). In 7 of these 8 cases, the plaque protrusion or thrombus was stabilized with medication alone. However, the remaining case showed growth and migration of the plaque protrusion or thrombus when treated with medication alone, and therefore, required further endovascular treatment. We identified that a history of symptomatic cerebral infarction and plaques with ulceration were risk factors for plaque protrusion or thrombus formation after CAS, and pre dilatation can decrease the risk of these complications. Medication was effective in most cases of plaque protrusion or thrombus; however, further endovascular treatment was required when medication alone was unsuccessful. PMID:25746309

  8. The actin regulators Enabled and Diaphanous direct distinct protrusive behaviors in different tissues during Drosophila development.

    PubMed

    Nowotarski, Stephanie H; McKeon, Natalie; Moser, Rachel J; Peifer, Mark

    2014-10-15

    Actin-based protrusions are important for signaling and migration during development and homeostasis. Defining how different tissues in vivo craft diverse protrusive behaviors using the same genomic toolkit of actin regulators is a current challenge. The actin elongation factors Diaphanous and Enabled both promote barbed-end actin polymerization and can stimulate filopodia in cultured cells. However, redundancy in mammals and Diaphanous' role in cytokinesis limited analysis of whether and how they regulate protrusions during development. We used two tissues driving Drosophila dorsal closure--migratory leading-edge (LE) and nonmigratory amnioserosal (AS) cells--as models to define how cells shape distinct protrusions during morphogenesis. We found that nonmigratory AS cells produce filopodia that are morphologically and dynamically distinct from those of LE cells. We hypothesized that differing Enabled and/or Diaphanous activity drives these differences. Combining gain- and loss-of-function with quantitative approaches revealed that Diaphanous and Enabled each regulate filopodial behavior in vivo and defined a quantitative "fingerprint"--the protrusive profile--which our data suggest is characteristic of each actin regulator. Our data suggest that LE protrusiveness is primarily Enabled driven, whereas Diaphanous plays the primary role in the AS, and reveal each has roles in dorsal closure, but its robustness ensures timely completion in their absence.

  9. Modeling the Excess Cell Surface Stored in a Complex Morphology of Bleb-Like Protrusions

    PubMed Central

    Wessler, Timothy; Yang, Xiaofeng; Chen, Alex; Roach, Nathan; Elston, Timothy C.; Wang, Qi; Jacobson, Ken; Forest, M. Gregory

    2016-01-01

    Cells transition from spread to rounded morphologies in diverse physiological contexts including mitosis and mesenchymal-to-amoeboid transitions. When these drastic shape changes occur rapidly, cell volume and surface area are approximately conserved. Consequently, the rounded cells are suddenly presented with a several-fold excess of cell surface whose area far exceeds that of a smooth sphere enclosing the cell volume. This excess is stored in a population of bleb-like protrusions (BLiPs), whose size distribution is shown by electron micrographs to be skewed. We introduce three complementary models of rounded cell morphologies with a prescribed excess surface area. A 2D Hamiltonian model provides a mechanistic description of how discrete attachment points between the cell surface and cortex together with surface bending energy can generate a morphology that satisfies a prescribed excess area and BLiP number density. A 3D random seed-and-growth model simulates efficient packing of BLiPs over a primary rounded shape, demonstrating a pathway for skewed BLiP size distributions that recapitulate 3D morphologies. Finally, a phase field model (2D and 3D) posits energy-based constitutive laws for the cell membrane, nematic F-actin cortex, interior cytosol, and external aqueous medium. The cell surface is equipped with a spontaneous curvature function, a proxy for the cell surface-cortex couple, that is a priori unknown, which the model “learns” from the thin section transmission electron micrograph image (2D) or the “seed and growth” model image (3D). Converged phase field simulations predict self-consistent amplitudes and spatial localization of pressure and stress throughout the cell for any posited stationary morphology target and cell compartment constitutive properties. The models form a general framework for future studies of cell morphological dynamics in a variety of biological contexts. PMID:27015526

  10. Self-organization of waves and pulse trains by molecular motors in cellular protrusions

    PubMed Central

    Yochelis, A.; Ebrahim, S.; Millis, B.; Cui, R.; Kachar, B.; Naoz, M.; Gov, N. S.

    2015-01-01

    Actin-based cellular protrusions are an ubiquitous feature of cells, performing a variety of critical functions ranging from cell-cell communication to cell motility. The formation and maintenance of these protrusions relies on the transport of proteins via myosin motors, to the protrusion tip. While tip-directed motion leads to accumulation of motors (and their molecular cargo) at the protrusion tip, it is observed that motors also form rearward moving, periodic and isolated aggregates. The origins and mechanisms of these aggregates, and whether they are important for the recycling of motors, remain open puzzles. Motivated by novel myosin-XV experiments, a mass conserving reaction-diffusion-advection model is proposed. The model incorporates a non-linear cooperative interaction between motors, which converts them between an active and an inactive state. Specifically, the type of aggregate formed (traveling waves or pulse-trains) is linked to the kinetics of motors at the protrusion tip which is introduced by a boundary condition. These pattern selection mechanisms are found not only to qualitatively agree with empirical observations but open new vistas to the transport phenomena by molecular motors in general. PMID:26335545

  11. Local protrusions formed on Si(111) surface by surface melting and solidification under applied tensile stress

    NASA Astrophysics Data System (ADS)

    Nishimura, T.; Tomitori, M.

    2016-09-01

    The surface structure and composition of Si(111) was modified, by heating it to 1300 °C in ultrahigh vacuum under an external tensile stress. A stress of approximately 1 GPa was applied, by pressing on the center of the rear side of the sample. This process produced two protrusions of approximately 100 μm in height, to the left and right of the center. Scanning Auger electron spectroscopy revealed Fe, Cr, Ni, and C impurities at the top of one protrusion, and C at the top of the other. These impurities likely diffused into the tops of the protrusions during heating, and segregated to the local surface during cooling when the protrusions formed. The protrusion formation mechanism is discussed. Their formation was related to non-uniform surface temperature, electromigration, piezoresistivity, freezing-point depression due to surface alloying with the impurities, and volume expansion during solidification from surface melting. These findings provide a perspective on controlling surface structures and compositions using heat and stress to induce self-assembly.

  12. Self-organization of waves and pulse trains by molecular motors in cellular protrusions.

    PubMed

    Yochelis, A; Ebrahim, S; Millis, B; Cui, R; Kachar, B; Naoz, M; Gov, N S

    2015-01-01

    Actin-based cellular protrusions are an ubiquitous feature of cells, performing a variety of critical functions ranging from cell-cell communication to cell motility. The formation and maintenance of these protrusions relies on the transport of proteins via myosin motors, to the protrusion tip. While tip-directed motion leads to accumulation of motors (and their molecular cargo) at the protrusion tip, it is observed that motors also form rearward moving, periodic and isolated aggregates. The origins and mechanisms of these aggregates, and whether they are important for the recycling of motors, remain open puzzles. Motivated by novel myosin-XV experiments, a mass conserving reaction-diffusion-advection model is proposed. The model incorporates a non-linear cooperative interaction between motors, which converts them between an active and an inactive state. Specifically, the type of aggregate formed (traveling waves or pulse-trains) is linked to the kinetics of motors at the protrusion tip which is introduced by a boundary condition. These pattern selection mechanisms are found not only to qualitatively agree with empirical observations but open new vistas to the transport phenomena by molecular motors in general. PMID:26335545

  13. Bleb Nucleation through Membrane Peeling

    NASA Astrophysics Data System (ADS)

    Alert, Ricard; Casademunt, Jaume

    2016-02-01

    We study the nucleation of blebs, i.e., protrusions arising from a local detachment of the membrane from the cortex of a cell. Based on a simple model of elastic linkers with force-dependent kinetics, we show that bleb nucleation is governed by membrane peeling. By this mechanism, the growth or shrinkage of a detached membrane patch is completely determined by the linker kinetics, regardless of the energetic cost of the detachment. We predict the critical nucleation radius for membrane peeling and the corresponding effective energy barrier. These may be typically smaller than those predicted by classical nucleation theory, implying a much faster nucleation. We also perform simulations of a continuum stochastic model of membrane-cortex adhesion to obtain the statistics of bleb nucleation times as a function of the stress on the membrane. The determinant role of membrane peeling changes our understanding of bleb nucleation and opens new directions in the study of blebs.

  14. PAK–PIX interactions regulate adhesion dynamics and membrane protrusion to control neurite outgrowth

    PubMed Central

    Santiago-Medina, Miguel; Gregus, Kelly A.; Gomez, Timothy M.

    2013-01-01

    Summary The roles of P21-activated kinase (PAK) in the regulation of axon outgrowth downstream of extracellular matrix (ECM) proteins are poorly understood. Here we show that PAK1–3 and PIX are expressed in the developing spinal cord and differentially localize to point contacts and filopodial tips within motile growth cones. Using a specific interfering peptide called PAK18, we found that axon outgrowth is robustly stimulated on laminin by partial inhibition of PAK–PIX interactions and PAK function, whereas complete inhibition of PAK function stalls axon outgrowth. Furthermore, modest inhibition of PAK–PIX stimulates the assembly and turnover of growth cone point contacts, whereas strong inhibition over-stabilizes adhesions. Point mutations within PAK confirm the importance of PIX binding. Together our data suggest that regulation of PAK–PIX interactions in growth cones controls neurite outgrowth by influencing the activity of several important mediators of actin filament polymerization and retrograde flow, as well as integrin-dependent adhesion to laminin. PMID:23321640

  15. Unusual extraction treatment of Class I bialveolar protrusion using microimplant anchorage.

    PubMed

    Chae, Jong-Moon

    2007-03-01

    This case report describes the treatment of a 16-year-old girl who had severe bialveolar protrusion. Patients with bialveolar protrusion are commonly treated with four first premolar extractions and retraction of the anterior teeth. Unfortunately in this patient, the mandibular left second molar had to be extracted because of extensive caries. To create sufficient space for retraction of the anterior teeth, the mandibular left posterior teeth were retracted with the mandibular posterior microimplant (1.2 mm in diameter, 6 mm long) placed into the retromolar area followed by en masse retraction of the mandibular anterior teeth. Microimplants can provide anchorage for obtaining a good facial profile even without premolar extraction in the case of bialveolar protrusion with the absence of the second molar.

  16. The dynamic distribution of fluorescent analogues of actin and myosin in protrusions at the leading edge of migrating Swiss 3T3 fibroblasts

    PubMed Central

    1988-01-01

    The formation of protrusions at the leading edge of the cell is an essential step in fibroblast locomotion. Using fluorescent analogue cytochemistry, ratio imaging, multiple parameter analysis, and fluorescence photobleaching recovery, the distribution of actin and myosin was examined in the same protrusions at the leading edge of live, locomoting cells during wound-healing in vitro. We have previously defined two temporal stages of the formation of protrusions: (a) initial protrusion and (b) established protrusion (Fisher et al., 1988). Actin was slightly concentrated in initial protrusions, while myosin was either totally absent or present at extremely low levels at the base of the initial protrusions. In contrast, established protrusions contained diffuse actin and actin microspikes, as well as myosin in both diffuse and structured forms. Actin and myosin were also localized along concave transverse fibers near the base of initial and established protrusions. The dynamics of myosin penetration into a relatively stable, established protrusion was demonstrated by recording sequential images over time. Myosin was shown to be absent from an initial protrusion, but diffuse and punctate myosin was detected in the same protrusion within 1-2 min. Fluorescence photobleaching recovery indicated that myosin was 100% immobile in the region behind the leading edge containing transverse fibers, in comparison to the 21% immobile fraction detected in the perinuclear region. Possible explanations of the delayed penetration of myosin into established protrusions and the implications on the mechanism of protrusion are discussed. PMID:3204122

  17. Investigating cytoskeletal function in chloroplast protrusion formation in the arctic-alpine plant Oxyria digyna.

    PubMed

    Holzinger, A; Wasteneys, G O; Lütz, C

    2007-05-01

    Arctic and alpine plants like Oxyria digyna have to face enhanced environmental stress. This study compared leaves from Oxyria digyna collected in the Arctic at Svalbard (78 degrees N) and in the Austrian Alps (47 degrees N) at cellular, subcellular, and ultrastructural levels. Oxyria digyna plants collected in Svalbard had significantly thicker leaves than the samples collected in the Austrian Alps. This difference was generated by increased thickness of the palisade and spongy mesophyll layers in the arctic plants, while epidermal cells had no significant size differences between the two habitats. A characteristic feature of arctic, alpine, and cultivated samples was the occurrence of broad stroma-filled chloroplast protrusions, 2 - 5 microm broad and up to 5 microm long. Chloroplast protrusions were in close spatial contact with other organelles including mitochondria and microbodies. Mitochondria were also present in invaginations of the chloroplasts. A dense network of cortical microtubules found in the mesophyll cells suggested a potential role for microtubules in the formation and function of chloroplast protrusions. No direct interactions between microtubules and chloroplasts, however, were observed and disruption of the microtubule arrays with the anti-microtubule agent oryzalin at 5 - 10 microM did not alter the appearance or dynamics of chloroplast protrusions. These observations suggest that, in contrast to studies on stromule formation in Nicotiana, microtubules are not involved in the formation and morphology of chloroplast protrusions in Oxyria digyna. The actin microfilament-disrupting drug latrunculin B (5 - 10 microM for 2 h) arrested cytoplasmic streaming and altered the cytoplasmic integrity of mesophyll cells. However, at the ultrastructural level, stroma-containing, thylakoid-free areas were still visible, mostly at the concave sides of the chloroplasts. As chloroplast protrusions were frequently found to be mitochondria-associated in Oxyria

  18. Treatment of dentoskeletal bimaxillary protrusion: additional extraction of maxillary second molars.

    PubMed

    Iijima, Shigeki; Yoshida, Mitsuru; Terada, Kazuto

    2009-07-01

    This case report describes the orthodontic treatment of an 18-year-old female patient with Class II malocclusion with dentoskeletal bimaxillary protrusion. An acceptable treatment result was obtained with a four first premolar extraction and an additional maxillary two second molar extraction plan. Standard edgewise appliances were placed after the premolars were extracted. A positive soft tissue response to treatment was achieved, and the patient's profile was improved, with a reduction of lip protrusion and mentalis strain. Dentally, the interincisal angle improved significantly as both the maxillary and mandibular incisors were uprighted after space closure. The occlusion remained stable during 2 years of follow-up.

  19. Open bite, dental alveolar protrusion, class I malocclusion: A successful treatment result.

    PubMed

    Alexander, C D

    1999-11-01

    This case report describes the treatment of a dentoalveolar protrusion, Class I malocclusion with an anterior open bite. The 21-year-old woman presented with a significant anteroposterior and vertical skeletal discrepancy. Her face was convex with procumbent lips. Intraorally, she had an anterior open bite of 4 mm, mild crowding, and an overjet of 4.5 mm. First premolar extractions in conjunction with tongue therapy and high pull headgear were used to reduce protrusion and close anterior open bite. Modification of a tongue thrust habit allowed conventional orthodontic treatment to correct this significant malocclusion and provide stability over the last 4 years.

  20. RhoD activated by fibroblast growth factor induces cytoneme-like cellular protrusions through mDia3C

    PubMed Central

    Koizumi, Kazuhisa; Takano, Kazunori; Kaneyasu, Akiko; Watanabe-Takano, Haruko; Tokuda, Emi; Abe, Tomoyuki; Watanabe, Naoki; Takenawa, Tadaomi; Endo, Takeshi

    2012-01-01

    The small GTPase RhoD regulates actin cytoskeleton to collapse actin stress fibers and focal adhesions, resulting in suppression of cell migration and cytokinesis. It also induces alignment of early endosomes along actin filaments and reduces their motility. We show here that a constitutively activated RhoD generated two types of actin-containing thin peripheral cellular protrusions distinct from Cdc42-induced filopodia. One was longer, almost straight, immotile, and sensitive to fixation, whereas the other was shorter, undulating, motile, and resistant to fixation. Moreover, cells expressing wild-type RhoD extended protrusions toward fibroblast growth factor (FGF) 2/4/8–coated beads. Stimulation of wild-type RhoD-expressing cells with these FGFs also caused formation of cellular protrusions. Nodules moved through the RhoD-induced longer protrusions, mainly toward the cell body. Exogenously expressed FGF receptor was associated with these moving nodules containing endosome-like vesicles. These results suggest that the protrusions are responsible for intercellular communication mediated by FGF and its receptor. Accordingly, the protrusions are morphologically and functionally equivalent to cytonemes. RhoD was activated by FGF2/4/8. Knockdown of RhoD interfered with FGF-induced protrusion formation. Activated RhoD specifically bound to mDia3C and facilitated actin polymerization together with mDia3C. mDia3C was localized to the tips or stems of the protrusions. In addition, constitutively activated mDia3C formed protrusions without RhoD or FGF stimulation. Knockdown of mDia3 obstructed RhoD-induced protrusion formation. These results imply that RhoD activated by FGF signaling forms cytoneme-like protrusions through activation of mDia3C, which induces actin filament formation. PMID:23034183

  1. Effect of Tongue Exercise on Protrusive Force and Muscle Fiber Area in Aging Rats

    ERIC Educational Resources Information Center

    Connor, Nadine P.; Russell, John A.; Wang, Hao; Jackson, Michelle A.; Mann, Laura; Kluender, Keith

    2009-01-01

    Purpose: Age-related changes in tongue function may contribute to dysphagia in elderly people. The authors' purpose was to investigate whether aged rats that have undergone tongue exercise would manifest increased protrusive tongue forces and increased genioglossus (GG) muscle fiber cross-sectional areas. Method: Forty-eight young adult,…

  2. Effects of tongue position and lung volume on voluntary maximal tongue protrusion force in humans.

    PubMed

    Saboisky, Julian P; Luu, Billy L; Butler, Jane E; Gandevia, Simon C

    2015-01-15

    Maximal voluntary protrusion force of the human tongue has not been examined in positions beyond the incisors or at different lung volumes. Tongue force was recorded with the tongue tip at eight positions relative to the incisors (12 and 4mm protrusion, neutral and 4, 12, 16, 24 and 32mm retraction) at functional residual capacity (FRC), total lung capacity (TLC) and residual volume (RV) in 15 healthy subjects. Maximal force occurred between 12mm and 32mm retraction (median 16mm). Maximum force at FRC was reproducible at the optimal tongue position across sessions (P=0.68). Across all positions at FRC the average force was highest at 24mm retraction (28.3±5.3N, mean±95% CI) and lowest at 12mm protrusion (49.1±4.6% maximum; P<0.05). Across all tongue positions, maximal force was on average 9.3% lower at FRC than TLC and RV (range: 4.5-12.7% maximum, P<0.05). Retracted positions produce higher-force protrusions with a small effect of lung volume.

  3. The cell ratchet: Interplay between efficient protrusions and adhesion determines cell motion

    PubMed Central

    Caballero, David; Voituriez, Raphaël; Riveline, Daniel

    2015-01-01

    Many physiological and pathological processes involve directed cell motion. In general, migrating cells are represented with a polarized morphology with extending and retracting protrusions at the leading edge. However, cell motion is a more complex phenomenon. Cells show heterogeneous morphologies and high protrusive dynamics is not always related to cell shape. This prevents the quantitative prediction of cell motion and the identification of cellular mechanisms setting directionality. Here we discuss the importance of protrusion fluctuations in directed cell motion. We show how their spatiotemporal distribution and dynamics determine the fluctuations and directions of cell motion for NIH3T3 fibroblasts plated on micro-patterned adhesive ratchets.1 We introduce efficient protrusions and direction index which capture short-term cell motility over hours: these new read-outs allow the prediction of parameters characteristic for the long-term motion of cells over days. The results may have important implications for the study of biological phenomena where directed cell migration is involved, in morphogenesis and in cancer. PMID:26313125

  4. Src and cortactin promote lamellipodia protrusion and filopodia formation and stability in growth cones

    PubMed Central

    He, Yingpei; Ren, Yuan; Wu, Bingbing; Decourt, Boris; Lee, Aih Cheun; Taylor, Aaron; Suter, Daniel M.

    2015-01-01

    Src tyrosine kinases have been implicated in axonal growth and guidance; however, the underlying cellular mechanisms are not well understood. Specifically, it is unclear which aspects of actin organization and dynamics are regulated by Src in neuronal growth cones. Here, we investigated the function of Src2 and one of its substrates, cortactin, in lamellipodia and filopodia of Aplysia growth cones. We found that up-regulation of Src2 activation state or cortactin increased lamellipodial length, protrusion time, and actin network density, whereas down-regulation had opposite effects. Furthermore, Src2 or cortactin up-regulation increased filopodial density, length, and protrusion time, whereas down-regulation promoted lateral movements of filopodia. Fluorescent speckle microscopy revealed that rates of actin assembly and retrograde flow were not affected in either case. In summary, our results support a model in which Src and cortactin regulate growth cone motility by increasing actin network density and protrusion persistence of lamellipodia by controlling the state of actin-driven protrusion versus retraction. In addition, both proteins promote the formation and stability of actin bundles in filopodia. PMID:26224308

  5. Effects of tongue position and lung volume on voluntary maximal tongue protrusion force in humans.

    PubMed

    Saboisky, Julian P; Luu, Billy L; Butler, Jane E; Gandevia, Simon C

    2015-01-15

    Maximal voluntary protrusion force of the human tongue has not been examined in positions beyond the incisors or at different lung volumes. Tongue force was recorded with the tongue tip at eight positions relative to the incisors (12 and 4mm protrusion, neutral and 4, 12, 16, 24 and 32mm retraction) at functional residual capacity (FRC), total lung capacity (TLC) and residual volume (RV) in 15 healthy subjects. Maximal force occurred between 12mm and 32mm retraction (median 16mm). Maximum force at FRC was reproducible at the optimal tongue position across sessions (P=0.68). Across all positions at FRC the average force was highest at 24mm retraction (28.3±5.3N, mean±95% CI) and lowest at 12mm protrusion (49.1±4.6% maximum; P<0.05). Across all tongue positions, maximal force was on average 9.3% lower at FRC than TLC and RV (range: 4.5-12.7% maximum, P<0.05). Retracted positions produce higher-force protrusions with a small effect of lung volume. PMID:25481541

  6. Feeding mechanisms in carp: crossflow filtration, palatal protrusions and flow reversals.

    PubMed

    Callan, W Todd; Sanderson, S Laurie

    2003-03-01

    It has been hypothesized that, when engulfing food mixed with inorganic particles during benthic feeding, cyprinid fish use protrusions of tissue from the palatal organ to retain the food particles while the inorganic particles are expelled from the opercular slits. In crossflow filtration, the particle suspension is pumped parallel to the filter surface as filtrate exits through the filter pores, causing the suspension to become more concentrated as it travels downstream along the filter. We used high-speed video endoscopy to determine whether carp Cyprinus carpio use crossflow filtration and/or palatal protrusions during benthic feeding. We found that carp use crossflow filtration to concentrate small food particles in the pharyngeal cavity while expelling small dense inorganic particles through the opercular slits and via spits. Our results suggest that, during feeding on small food particles, palatal protrusions serve a localized chemosensory function rather than a mechanical particle-sorting function. However, palatal protrusions did retain large food particles while large inorganic particles were spit anteriorly from the mouth. We also investigated whether flow is continuous and unidirectional during suspension feeding in carp. As reported previously for ventilation in hedgehog skates and for certain industrial crossflow filtration applications, we observed that flow is pulsatile and bidirectional during feeding. These results have implications for hydrodynamic models of crossflow filtration in suspension-feeding fishes.

  7. Imitation of Tongue Protrusion in Human Neonates: Specificity of the Response in a Large Sample

    ERIC Educational Resources Information Center

    Nagy, Emese; Pilling, Karen; Orvos, Hajnalka; Molnar, Peter

    2013-01-01

    Although a large body of evidence has accumulated on the young human infant's ability to imitate, the phenomenon has failed to gain unanimous acceptance. Imitation of tongue protrusion, the most tested gesture to date, was examined in a sample of 115 newborns in the first 5 days of life in 3 seating positions. An ethologically based…

  8. Exo70 Generates Membrane Curvature for Morphogenesis and Cell Migration

    PubMed Central

    Zhao, Yuting; Liu, Jianglan; Yang, Changsong; Capraro, Benjamin R.; Baumgart, Tobias; Bradley, Ryan P.; Ramakrishnan, N.; Xu, Xiaowei; Radhakrishnan, Ravi; Svitkina, Tatyana; Guo, Wei

    2013-01-01

    Dynamic shape changes of the plasma membrane are fundamental to many processes ranging from morphogenesis and cell migration to phagocytosis and viral propagation. Here we demonstrate that Exo70, a component of the exocyst complex, induces tubular membrane invaginations towards the lumen of synthetic vesicles in vitro and generates protrusions on the surface of cells. Biochemical analyses using Exo70 mutants and independent molecular dynamics simulations based on Exo70 structure demonstrate that Exo70 generates negative membrane curvature through an oligomerization-based mechanism. In cells, the membrane-deformation function of Exo70 is required for protrusion formation and directional cell migration. Exo70 thus represents a membrane-bending protein that may couple actin dynamics and plasma membrane remodeling for morphogenesis. PMID:23948253

  9. Electrokinetic effects near a membrane

    NASA Astrophysics Data System (ADS)

    Lacoste, David

    2009-03-01

    We discuss the electrostatic and electrokinetic contribution to the elastic moduli of a cell or artificial membrane placed in an electrolyte and driven by a DC electric field. The field drives ion currents across the membrane, through specific channels, pumps or natural pores. In steady state, charges accumulate in the Debye layers close to the membrane, modifying the membrane elastic moduli. We first study a model of a membrane of zero thickness, later generalizing this treatment to allow for a finite thickness and finite dielectric constant. Our results clarify and extend the results presented in [D. Lacoste, M. Cosentino Lagomarsino, and J. F. Joanny, Europhys. Lett., 77, 18006 (2007)], by providing a physical explanation for a destabilizing term proportional to kps^3 in the fluctuation spectrum, which we relate to a nonlinear (E^2) electro-kinetic effect called induced-charge electro-osmosis (ICEO). Recent studies of ICEO have focused on electrodes and polarizable particles, where an applied bulk field is perturbed by capacitive charging of the double layer and drives flow along the field axis toward surface protrusions; we predict similar ICEO flows around driven membranes, due to curvature-induced tangential fields within a non-equilibrium double layer, which hydrodynamically enhance protrusions.

  10. Premaxillary movements in cyprinodontiform fishes: an unusual protrusion mechanism facilitates "picking" prey capture.

    PubMed

    Ferry-Graham, Lara A; Gibb, Alice C; Hernandez, L Patricia

    2008-01-01

    Premaxillary protrusion is hypothesized to confer a number of feeding advantages to teleost fishes; however, most proposed advantages relate to enhanced stealth or suction production during prey capture. Cyprinodontiformes exhibit an unusual form of premaxillary protrusion where the descending process of the premaxilla does not rotate anteriorly to occlude the sides of the open mouth during prey capture. Instead, the premaxilla is protruded such that it gives the impression of a beak during prey capture. We quantified premaxillary kinematics during feeding in four cyprinodontiform taxa and compared them with three percomorph taxa to identify any performance consequences of this protrusion mechanism. Individual prey capture events were recorded using digital high-speed video at 250-500 frames per second (n >or= 4 individuals, >or= 4 strikes per individual). Species differed in the timing of movement and the maximum displacement of the premaxilla during the gape cycle and in the contribution of the premaxilla to jaw closing. Cyprinodontiform taxa produced less premaxillary protrusion than the percomorph taxa, and were consistently slower in the time to maximum gape. Further, it appears cyprinodontiforms can alter the contribution of the premaxilla to mouth closure on an event-specific basis. We were able to demonstrate that, within at least one species, this variability is associated with the location of the prey (bottom vs. water column). Cyprinodontiform upper jaw movements likely reflect increased dexterity associated with a foraging ecology where prey items are "picked" from a variety of locations: the bottom, water column, or surface. We postulate that dexterity requires slow, precisely controlled jaw movements; thus, may be traded off for some aspects of suction-feeding performance, such as protrusion distance and speed. PMID:18619823

  11. Early carbon mobilization and radicle protrusion in maize germination.

    PubMed

    Sánchez-Linares, Luis; Gavilanes-Ruíz, Marina; Díaz-Pontones, David; Guzmán-Chávez, Fernando; Calzada-Alejo, Viridiana; Zurita-Villegas, Viridiana; Luna-Loaiza, Viridiana; Moreno-Sánchez, Rafael; Bernal-Lugo, Irma; Sánchez-Nieto, Sobeida

    2012-07-01

    Considerable amounts of information is available on the complex carbohydrates that are mobilized and utilized by the seed to support early seedling development. These events occur after radicle has protruded from the seed. However, scarce information is available on the role of the endogenous soluble carbohydrates from the embryo in the first hours of germination. The present work analysed how the soluble carbohydrate reserves in isolated maize embryos are mobilized during 6-24 h of water imbibition, an interval that exclusively embraces the first two phases of the germination process. It was found that sucrose constitutes a very significant reserve in the scutellum and that it is efficiently consumed during the time in which the adjacent embryo axis is engaged in an active metabolism. Sucrose transporter was immunolocalized in the scutellum and in vascular elements. In parallel, a cell-wall invertase activity, which hydrolyses sucrose, developed in the embryo axis, which favoured higher glucose uptake. Sucrose and hexose transporters were active in the embryo tissues, together with the plasma membrane H(+)-ATPase, which was localized in all embryo regions involved in both nutrient transport and active cell elongation to support radicle extension. It is proposed that, during the initial maize germination phases, a net flow of sucrose takes place from the scutellum towards the embryo axis and regions that undergo elongation. During radicle extension, sucrose and hexose transporters, as well as H(+)-ATPase, become the fundamental proteins that orchestrate the transport of nutrients required for successful germination.

  12. Early carbon mobilization and radicle protrusion in maize germination

    PubMed Central

    Sánchez-Linares, Luis; Gavilanes-Ruíz, Marina; Díaz-Pontones, David; Guzmán-Chávez, Fernando; Calzada-Alejo, Viridiana; Zurita-Villegas, Viridiana; Luna-Loaiza, Viridiana; Moreno-Sánchez, Rafael; Bernal-Lugo, Irma; Sánchez-Nieto, Sobeida

    2012-01-01

    Considerable amounts of information is available on the complex carbohydrates that are mobilized and utilized by the seed to support early seedling development. These events occur after radicle has protruded from the seed. However, scarce information is available on the role of the endogenous soluble carbohydrates from the embryo in the first hours of germination. The present work analysed how the soluble carbohydrate reserves in isolated maize embryos are mobilized during 6–24 h of water imbibition, an interval that exclusively embraces the first two phases of the germination process. It was found that sucrose constitutes a very significant reserve in the scutellum and that it is efficiently consumed during the time in which the adjacent embryo axis is engaged in an active metabolism. Sucrose transporter was immunolocalized in the scutellum and in vascular elements. In parallel, a cell-wall invertase activity, which hydrolyses sucrose, developed in the embryo axis, which favoured higher glucose uptake. Sucrose and hexose transporters were active in the embryo tissues, together with the plasma membrane H+-ATPase, which was localized in all embryo regions involved in both nutrient transport and active cell elongation to support radicle extension. It is proposed that, during the initial maize germination phases, a net flow of sucrose takes place from the scutellum towards the embryo axis and regions that undergo elongation. During radicle extension, sucrose and hexose transporters, as well as H+-ATPase, become the fundamental proteins that orchestrate the transport of nutrients required for successful germination. PMID:22611232

  13. Effect of microscale protrusions on local fluid flow and mass transport in the presence of forced convection

    SciTech Connect

    Matzen, G.W.

    1997-01-01

    Three-dimensional creeping flow around single, axisymmetric protrusions is studied numerically using the boundary-integral technique. Emphasis is placed upon cylindrical protrusions on plane walls for various height-to-radius (h-to-a) aspect ratios, but cones and sections of spheres protruding from plane walls are also briefly examined. The presented items include shear-stress distributions, shear-stress contours, extents of the fluid-flow disturbance, total forces and torques on the cylinders, streamlines, and skin-friction lines. Also included is a discussion of flow topology around axisymmetric geometries. No flow reversal is observed for cylindrical protrusions with aspect ratios greater than 2.4 to 2.6. At higher aspect ratios, the fluid tends to be swept around cylindrical protrusions with little vertical motion. At lower aspect ratios, the strength of the recirculation increases, and the recirculation region becomes wider in the transverse direction and narrower in the flow direction. Also, the recirculation pattern begins to resemble the closed streamline patterns in two-dimensional flow over square ridges. However, unlike two-dimensional flow, closed streamline patterns are not observed. For arbitrary axisymmetric geometries, the extent of the fluid-flow disturbance can be estimated with the total force that is exerted on the protrusion. When the same force is exerted on protrusions with different aspect ratios, the protrusion with the higher aspect ratio tends to have a greater disturbance in the flow direction and a smaller disturbance in the transverse direction. The total force exerted on cylindrical protrusions with rounded corners is only slightly lower than the total force exerted on cylindrical protrusions with sharp corners.

  14. Heterotypic and homotypic associations between ezrin and moesin, two putative membrane-cytoskeletal linking proteins.

    PubMed Central

    Gary, R; Bretscher, A

    1993-01-01

    Ezrin and moesin are components of actin-rich cell surface structures that are thought to function as membrane-cytoskeletal linking proteins. Here we show that a stable complex of ezrin and moesin can be isolated from cultured cells by immunoprecipitation with specific antibodies. The capacity of these two proteins to interact directly was confirmed with a blot-overlay procedure in which biotin-tagged proteins in solution were incubated with immobilized binding partners. In addition to the heterotypic association of ezrin and moesin, homotypic binding of ezrin to ezrin and of moesin to moesin was also demonstrated in vitro. These results suggest mechanisms by which ezrin and moesin might participate in dynamic aspects of cortical cytoskeletal structure. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 Fig. 5 PMID:8248180

  15. Cooperation between β- and γ-cytoplasmic actins in the mechanical regulation of endothelial microparticle formation.

    PubMed

    Latham, Sharissa L; Chaponnier, Christine; Dugina, Vera; Couraud, Pierre-Olivier; Grau, Georges E R; Combes, Valery

    2013-02-01

    Elevated endothelial microparticle (MP) levels are observed in numerous diseases, increasingly supporting roles as effectors and valuable markers of vascular dysfunction. While a contractile role for the actin cytoskeleton has been implicated in vesiculation, i.e., MP production, the precise interactions and mechanisms of its constituents, β- and γ-cytoplasmic actins, is unknown. Human cerebral microvascular endothelial cells were stimulated with known agonists, and vesiculation development was monitored by scanning electron microscopy (SEM) and flow cytometry. These data in combination provide new insight into the kinetics, patterns of vesiculating cell recruitment, and degrees of response specific to stimuli. Reorganization of β- and γ-actins, F-actin, vinculin, and talin accompanied significant MP release. β-Actin redistribution into basal stress fibers following stimulation was associated with increased apically situated actin-rich particulate structures, which in turn directly correlated with electron-lucent membrane protrusions observed by SEM. Y-27632 Rho-kinase inhibition abolished basal β-actin fiber formation, minimizing apically associated actin-rich structures, significantly reducing membrane protrusions and MP release to near basal levels. Cytoskeletal protein expression and distribution varied between MPs and mother cells, as determined by Western blot. These data strongly suggest that β-actin plays an active facilitative role in agonist-induced protuberance formation, through mechanical interactions with newly described actin-rich structures. PMID:23159932

  16. Treatment of Bi-maxillary Protrusion with Impacted Maxillary and Mandibular Canines: Case Report.

    PubMed

    Shdrma, Vipul Kumar; Yadav, Kirti; Nagar, Amit; Tandon, Pradeep; Chaturvedi, Thakur Prasad

    2016-01-01

    Bi-maxillary protrusion in adolescent patients has traditionally been treated by extraction of the four first premolars and retraction ofthe anterior teeth. The ectopic eruption of the maxiIlary permanent canines is a frequently encountered clinical problem. Orthodontic treatment of the impacted teeth remains a challenge for clinicians. If it is associated with other dental and skeletalproblems, there will be further complications to the treatment plan. In such cases, if we extract canines, then problems with this approach are restricted to anatomical and functional limitations ofpremolars substitution of canines. Here, we are presenting a case report of bi-maxillary dento-alveolar protrusion with the impacted maxillary and mandibular left canines and its management.

  17. [Coblation of nucleus pulposus in treatment of military men's lumbar disc protrusions].

    PubMed

    Manukovskiĭ, V A; Badalov, V I; Tiulikov, K V; Korostelev, K E

    2012-06-01

    Mini-invasive surgery is used more often for the treatment of spinal disc herniations today. Coblation is one of such contemporary methods. The aim of our investigation was to evaluate results of percutaneous disc nucleoplasty using coblation. 35 patients with disc protrusions diagnosed by MRI underwent operations using this method. We used Visual Analogue Pain Scale, took into account time of painless sitting, standing and walking position, and also patients' satisfaction to assess the immediate results of treatment and short-time outcomes. It was noted the better immediate results in group of operated patients in comparison with the non-operated group. It was obvious either on the table or the day after operation. There were 84 and 93% of good results in 3 months and in 1 year after operation, respectively. Coblation is definitely modern and effective way to treat disc protrusions with different types, sizes, and may be used in lumbar spine wherever.

  18. [Mecring threaded cup in total arthroplasty of the hip with acetabular protrusion].

    PubMed

    Bednarek, A; Blacha, J; Gagała, J

    1998-01-01

    Results of total arthroplasty of the hip with acetabular protrusion with the use of Mecring cup in 21 patients (25 hips) aged 25-75 years (mean 62) are reported. Morsalised impacted autografts were used to fill acetabular defect in all cases. There were 14 rheumatoid arthritis patients, 4 cases of osteoarthritis and 3 cases of Otto-Chrobak disease. Mean follow-up was 2.5 years (range 6 months-5 years). All autografts were incorporated after 6-9 months. Only one loosening and migration of the cup occurred. One patient died because of pulmonary embolism. Harris Hip Score increased from 37 points (22-49) preoperatively to 81 points (65-89) at the latest follow-up. Incorporation of autografts reinforced acetabular bottom and prevented recurrence of protrusion. Bone remodeling indicates restoring normal biomechanics of the hip.

  19. On the instability of conical protrusions of liquid surface in electric field

    SciTech Connect

    Petrin, A. B.

    2007-12-15

    Electric field is analyzed at the tip of the conical protrusion of a conductive liquid surface that forms as the field strength approaches certain threshold values. When the field exceeds the threshold, the cone tip generally becomes unstable and emits a slender jet. The field strength at the sharp tip of a conducting liquid cone tends to infinity. Even though the field has a singularity, the cone tip becomes unstable only if the semi-cone angle decreases below the Taylor angle, which is the same for all conducting liquids. It is shown that similar conical protrusions can form if the liquid is a dielectric and its permittivity is sufficiently high. In this case, the equilibrium cone angle depends on the permittivity. When the permittivity is lower than approximately 17.60, cones cannot form, and the ensuing phenomena do not occur.

  20. Human Newborns Match Tongue Protrusion of Disembodied Human and Robotic Mouths

    ERIC Educational Resources Information Center

    Soussignan, Robert; Courtial, Alexis; Canet, Pierre; Danon-Apter, Gisele; Nadel, Jacqueline

    2011-01-01

    No evidence had been provided so far of newborns' capacity to give a matching response to 2D stimuli. We report evidence from 18 newborns who were presented with three types of stimuli on a 2D screen. The stimuli were video-recorded displays of tongue protrusion shown by: (a) a human face, (b) a human tongue from a disembodied mouth, and (c) an…

  1. Friction-induced nanofabrication method to produce protrusive nanostructures on quartz

    NASA Astrophysics Data System (ADS)

    Song, Chenfei; Li, Xiaoying; Yu, Bingjun; Dong, Hanshan; Qian, Linmao; Zhou, Zhongrong

    2011-12-01

    In this paper, a new friction-induced nanofabrication method is presented to fabricate protrusive nanostructures on quartz surfaces through scratching a diamond tip under given normal loads. The nanostructures, such as nanodots, nanolines, surface mesas and nanowords, can be produced on the target surface by programming the tip traces according to the demanded patterns. The height of these nanostructures increases with the increase of the number of scratching cycles or the normal load. Transmission electron microscope observations indicated that the lattice distortion and dislocations induced by the mechanical interaction may have played a dominating role in the formation of the protrusive nanostructures on quartz surfaces. Further analysis reveals that during scratching, a contact pressure ranged from 0.4 P y to P y ( P y is the critical yield pressure of quartz) is apt to produce protuberant nanostructures on quartz under the given experimental conditions. Finally, it is of great interest to find that the protrusive nanostructures can be selectively dissolved in 20% KOH solution. Since the nanowords can be easily 'written' by friction-induced fabrication and 'erased' through selective etching on a quartz surface, this friction-induced method opens up new opportunities for future nanofabrication.

  2. Par3/Bazooka and phosphoinositides regulate actin protrusion formation during Drosophila dorsal closure and wound healing.

    PubMed

    Pickering, Karen; Alves-Silva, Juliana; Goberdhan, Deborah; Millard, Tom H

    2013-02-01

    Effective wound closure mechanisms are essential for maintenance of epithelial structure and function. The repair of wounded epithelia is primarily driven by the cells bordering the wound, which become motile after wounding, forming dynamic actin protrusions along the wound edge. The molecular mechanisms that trigger wound edge cells to become motile following tissue damage are not well understood. Using wound healing and dorsal closure in Drosophila, we identify a direct molecular link between changes in cell-cell adhesion at epithelial edges and induction of actin protrusion formation. We find that the scaffolding protein Par3/Bazooka and the lipid phosphatase Pten are specifically lost from cell-cell junctions at epithelial edges. This results in a localized accumulation of phosphatidylinositol 3,4,5-trisphosphate (PIP3), which promotes the formation of actin protrusions along the epithelial edge. Depleting PIP3 results in defective epithelial closure during both dorsal closure and wound healing. These data reveal a novel mechanism that directly couples loss of epithelial integrity to activation of epithelial closure.

  3. Numerical investigation of root canal irrigation adopting innovative needles with dimple and protrusion.

    PubMed

    Li, Ping; Zhang, Di; Xie, Yonghui; Lan, Jibing

    2013-01-01

    As important passive flow control methods, dimples and protrusions have been successfully implemented via geometric modifications to manipulate flow fields to get a desired flow parameters enhancement. In this research, two novel needles were proposed based on a prototype by means of the dimple and protrusion, and flow patterns within a root canal during final irrigation with these needles were numerically investigated. The calculation cases consistent with the clinically realistic irrigant flow rates, which are 0.02, 0.16 and 0.26 mL s(-1) are marked as case A, B and C, respectively. The characteristic parameters to estimate irrigation efficiency, such as shearing effect, mean apical pressure, irrigation replacement and fluid agitation, were compared and the optimal geometry in every calculation case was obtained. As shown from the results, flow rates and needle geometries were the causes of irrigation parameters variations. The sum of shear stress, irrigation replacement and fluid agitation were equal in the low flow rate case A, however, the needle with a protrusion on its tip had advantages in the three irrigation characteristic parameters above in calculation case B, and the needle with a dimple on its tip had advantages in calculation case C. Furthermore, the needles proposed did not give rise to the risk of irrigant extrusion. These needles can be better choices at larger flow rates. Therefore, needle geometry optimizations utilizing passive flow control methods are worthy to be investigated in the root canal irrigation enhancement. PMID:23957591

  4. Atmospheric effects on radiometry from zenith of a plane with dark vertical protrusions

    NASA Technical Reports Server (NTRS)

    Otterman, J.

    1983-01-01

    Effects of an optically thin plane-parallel scattering atmosphere on radiometric imaging from the zenith of a specific surface-type are analyzed. The surface model was previously developed to describe arid steppe, where the sparse vegetation forms dark vertical protrusions from the bright soil-plane. The analysis is in terms of the surface reflectivity to the zenith r sub p for the direct beam, which is formulated as r sub p = r sub i exp (-s tan theta sub 0), where v sub i is the Lambert law reflectivity of the soil, the protrusions parameters s is the projection on a vertical plane of protrusions per unit area and theta sub 0 is the zenith angle. The surface reflectivity r sub p is approximately equal to that for the global irradiance (which is directly measured in the field) only for a narrow range of the solar zenith angles. The effects of the atmosphere when imaging large uniform areas of this type are comparable to those in imaging a Lambert surface with a reflectivity r sub p. Thus, the effects can be approximated by those in the case of a dark Lambert surface (analyzed previously), inasmuch as r sub p is smaller than the soil reflectivity r sub i for any off-zenith illumination. The surface becomes effectively darker with increasing solar zenith angle. Adjacency effects of a reflection from one area and scattering in the instantaneous field of view (object pixel) are analyzed as cross radiance and cross irradiance.

  5. Autosomal recessive mental retardation syndrome with anterior maxillary protrusion and strabismus: MRAMS syndrome.

    PubMed

    Basel-Vanagaite, Lina; Rainshtein, Limor; Inbar, Dov; Gothelf, Doron; Hennekam, Raoul; Straussberg, Rachel

    2007-08-01

    We report on a family in whom the combination of mental retardation (MR), anterior maxillary protrusion, and strabismus segregates. The healthy, consanguineous parents (first cousins) of Israeli-Arab descent had 11 children, 7 of whom (5 girls) were affected. They all had severe MR. Six of the seven had anterior maxillary protrusion with vertical maxillary excess, open bite, and prominent crowded teeth. None of the sibs with normal intelligence had jaw or dental anomalies. The child with MR but without a jaw anomaly was somewhat less severely retarded, had seizures and severe psychosis, which may point to his having a separate disorder. Biochemical and neurological studies, including brain MRI and standard cytogenetic studies, yielded normal results; fragile X was excluded, no subtelomeric rearrangements were detectable, and X-inactivation studies in the mother showed random inactivation. We have been unable to find a similar disorder in the literature, and suggest that this is a hitherto unreported autosomal recessive disorder, which we propose to name MRAMS (mental retardation, anterior maxillary protrusion, and strabismus).

  6. [Validity of modified radiological views to detect screw protrusion at the distal radius. A comparative study with computerized tomography].

    PubMed

    Mora-Pascual, F E; Aguilella-Fernández, L

    2013-01-01

    Volar fixed-angle plates (VFAP) are currently widely used for the treatment of extra-articular distal radius fractures. Using these plates has a high risk of articular and dorsal screw protrusion due to their special configuration. The aim of this study is to assess the validity of the standard X-rays, performed with the help of wedged supports, in order to detect articular and dorsal screw protrusion. A comparison with computed tomography (CT) scan imaging has been made. The outcome of 26 patients with distal radius articular fracture, treated with a VFAP, is reported. Good correlation between modified X-rays and CT scan was observed. A sensitivity of 100% for articular protrusion and 66% for dorsal have been obtained. When detecting screw protrusion at the distal radius, the use of wedged supports to perform special X-rays intraoperatively is an effective tool.

  7. Coordination of protrusion and translocation of the keratocyte involves rolling of the cell body

    PubMed Central

    1996-01-01

    We have investigated the relationship between lamellipodium protrusion and forward translocation of the cell body in the rapidly moving keratocyte. It is first shown that the trailing, ellipsoidal cell body rotates during translocation. This was indicated by the rotation of the nucleus and the movement of cytoplasmic organelles, as well as of exogenously added beads used as markers. Activated or Con A-coated fluorescent beads that were overrun by cells were commonly endocytosed and rotated with the internal organelles. Alternatively, beads applied to the rear of the cell body via a micropipette adhered to the dorsal cell surface and also moved forward, indicating that both exterior and underlying cortical elements participated in rotation. Manipulation of keratocytes with microneedles demonstrated that pushing or restraining the cell body in the direction of locomotion, and squeezing it against the substrate, which temporarily increased the intracellular pressure, did not effect the rate of lamellipodium protrusion. Rotation and translocation of the cell body continued momentarily after arrest of lamellipodium protrusion by cytochalasin B, indicating that these processes were not directly dependent on actin polymerization. The cell body was commonly flanked by phase-dense "axles," extending from the cell body into the lamellipodium. Phalloidin staining showed these to be comprised of actin bundles that splayed forward into the flanks of the lamellipodium. Disruption of the bundles on one side of the nucleus by traumatic microinjection resulted in rapid retraction of the cell body in the opposite direction, indicating that the cell body was under lateral contractile stress. Myosin II, which colocalizes with the actin bundles, presumably provides the basis of tension generation across and traction of the cell body. We propose that the basis of coupling between lamellipodium protrusion and translocation of the cell body is a flow of actin filaments from the front

  8. EVALUATION OF MAGNETIC RESONANCE IMAGING GUIDELINES FOR DIFFERENTIATION BETWEEN THORACOLUMBAR INTERVERTEBRAL DISK EXTRUSIONS AND INTERVERTEBRAL DISK PROTRUSIONS IN DOGS.

    PubMed

    De Decker, Steven; Gomes, Sergio A; Packer, Rowena Ma; Kenny, Patrick J; Beltran, Elsa; Parzefall, Birgit; Fenn, Joe; Nair, Devi; Nye, George; Volk, Holger A

    2016-09-01

    Four MRI variables have recently been suggested to be independently associated with a diagnosis of thoracolumbar intervertebral disk extrusion or protrusion. Midline intervertebral disk herniation, and partial intervertebral disk degeneration were associated with intervertebral disk protrusion, while presence of a single intervertebral disk herniation and disk material dispersed beyond the boundaries of the intervertebral disk space were associated with intervertebral disk extrusion. The aim of this retrospective, cross-sectional study was to determine whether using these MRI variables improves differentiation between thoracolumbar intervertebral disk extrusions and protrusions. Eighty large breed dogs with surgically confirmed thoracolumbar intervertebral disk extrusions or protrusions were included. Randomized MRI studies were presented on two occasions to six blinded observers, which were divided into three experience categories. During the first assessment, observers made a presumptive diagnosis of thoracolumbar intervertebral disk extrusion or protrusion without guidelines. During the second assessment they were asked to make a presumptive diagnosis with the aid of guidelines. Agreement was evaluated by Kappa-statistics. Diagnostic accuracy significantly improved from 70.8 to 79.6% and interobserver agreement for making a diagnosis of intervertebral disk extrusion or intervertebral disk protrusion improved from fair (κ = 0.27) to moderate (κ = 0.41) after using the proposed guidelines. Diagnostic accuracy was significantly influenced by degree of observer experience. Intraobserver agreement for the assessed variables ranged from fair to excellent and interobserver agreement ranged from fair to moderate. The results of this study suggest that the proposed imaging guidelines can aid in differentiating thoracolumbar intervertebral disk extrusions from protrusions. PMID:27374979

  9. Determinants of successful treatment of bimaxillary protrusion: orthodontic treatment versus anterior segmental osteotomy.

    PubMed

    Baek, Seung-Hak; Kim, Byoung-Ho

    2005-03-01

    The purpose of this study was to investigate the differences in initial skeletal, dental, and soft tissue characteristics of bimaxillary protrusion (BP) patients to determine poor or good results with orthodontic treatment (OT) or anterior segmental osteotomy (ASO) with extraction of four first premolars. Lateral cephalometric radiographs of 46 adult Korean females with BP were analyzed before treatment (T0) and after treatment (T1). According to the measurements at T1, patients were classified into group 1 (poor result with OT, n = 12), group 2 (good result with OT, n = 11), group 3 (poor result with ASO, n = 5), and group 4 (good result with ASO, n = 18). Sagittal, vertical, dental, and soft tissue variables were measured. The differences at T0 among the four groups were compared by one way analysis of variance test and verified by Scheffe's multiple comparison test. Stepwise discriminant analysis was performed to find decisive predictors. Skeletal class II malocclusion tendency, less developed chin, and vertical facial growth pattern were related with group 1. Overly uprighted and less protrusive upper and lower incisor, near normal interincisal angle (IIA), less protrusive upper lip, and more obtuse lower nasolabial angle (NLA) were related with group 3. IIA, U1-NA distance, combination factor, interlabial gap, lower NLA, pterygomaxillary fissure-N, and posterior nasal spine-anterior nasal spine were selected as significant variables for discriminating the four groups. The percentage of correctly classified cases was 91.3%. In particular, the discriminant function showed the highest accuracy in the prediction of group 4. These variables and discriminant functions contributed to the differential diagnosis on BP to make a procedural decision between OT and ASO.

  10. Reversal of negative charges on the surface of Escherichia coli thioredoxin: pockets versus protrusions.

    PubMed

    Mancusso, Romina; Cruz, Eduardo; Cataldi, Marcela; Mendoza, Carla; Fuchs, James; Wang, Hsin; Yang, Xiaomin; Tasayco, María Luisa

    2004-04-01

    Recent studies of proteins with reversed charged residues have demonstrated that electrostatic interactions on the surface can contribute significantly to protein stability. We have used the approach of reversing negatively charged residues using Arg to evaluate the effect of the electrostatics context on the transition temperature (T(m)), the unfolding Gibbs free energy change (DeltaG), and the unfolding enthalpy change (DeltaH). We have reversed negatively charged residues at a pocket (Asp9) and protrusions (Asp10, Asp20, Glu85), all located in interconnecting segments between elements of secondary structure on the surface of Arg73Ala Escherichia coli thioredoxin. DSC measurements indicate that reversal of Asp in a pocket (Asp9Arg/Arg73Ala, DeltaT(m) = -7.3 degrees C) produces a larger effect in thermal stability than reversal at protrusions: Asp10Arg/Arg73Ala, DeltaT(m) = -3.1 degrees C, Asp20Arg/Arg73Ala, DeltaT(m) = 2.0 degrees C, Glu85Arg/Arg73Ala, DeltaT(m) = 3.9 degrees ). The 3D structure of thioredoxin indicates that Asp20 and Glu85 have no nearby charges within 8 A, while Asp9 does not only have Asp10 as sequential neighbor, but it also forms a 5-A long-range ion pair with the solvent-exposed Lys69. Further DSC measurements indicate that neutralization of the individual charges of the ion pair Asp9-Lys69 with nonpolar residues produces a significant decrease in stability in both cases: Asp9Ala/Arg73Ala, DeltaT(m) = -3.7 degrees C, Asp9Met/Arg73Ala, DeltaT(m) = -5.5 degrees C, Lys69Leu/Arg73Ala, DeltaT(m) = -5.1 degrees C. However, thermodynamic analysis shows that reversal or neutralization of Asp9 produces a 9-15% decrease in DeltaH, while both reversal of Asp at protrusions and neutralization of Lys69 produce negligible changes. These results correlate well with the NMR analysis, which demonstrates that only the substitution of Asp9 produces extensive conformational changes and these changes occur in the surroundings of Lys69. Our results led us to

  11. Albedo of a forest modeled as a plane with dense protrusions

    NASA Technical Reports Server (NTRS)

    Otterman, J.

    1984-01-01

    An analytical model for the absorption of solar radiation by surfaces such as a pine forest or a wheat field is presented. Objectives include understanding the parameters affecting the absorption of the solar irradiance in a complex structure, and determining the influence of the direction of illumination on light trapping. The surface is treated as a Lambertian reflectivity soil-plane; thin, vertical cylinders are regarded as Lambertian reflectors. Using a dimensionless protrusion parameter based on the height and diameter of the vertical plant elements, optical characteristics (e.g., the dependence of the albedo on the solar zenith angle) of a complex structure are well quantified.

  12. Infantile perianal pyramidal protrusion: a case report with dermoscopy and ultrasound findings

    PubMed Central

    Lamberti, Arianna; Filippou, Georgios; Adinolfi, Antonella; Fimiani, Michele; Rubegni, Pietro

    2015-01-01

    Infantile perianal pyramidal protrusion, it is a rare benign cutaneous condition described in relatively recent times. It is considered to be under-reported in the pediatric literature because it is often mistaken for other conditions. The unawareness of this lesion may be responsible for an excessive concern both in physician and in parents, which leads to overly aggressive and unnecessary treatments. Thus its recognition has many implications regarding proper management and treatment. We report a typical presentation of IPPP in which the diagnosis was based on the use of non-invasive diagnostic tools and in particular of dermoscopy and ultrasonography. PMID:26114069

  13. Crawling of a driven adherent membrane

    NASA Astrophysics Data System (ADS)

    Baumgaertner, A.

    2012-10-01

    We discuss motions of an elastic N × M membrane model whose constituents can bind reversibly with strength ɛ to adhesive sites of a flat substrate. One of the edges of the membrane ("front") is driven in one direction at rate constant p by N stochastically treadmilling short parallel lines ("cortex"). The main conclusions derived from Monte Carlo studies of this model are the following: (a) Since the polymerizing cortex pushes only the leading edge of the membrane, the major part of the membranes is dragged behind. Therefore, the locomotion of the membrane can be described by frictional sliding processes which are asymmetrically distributed between front and rear of the membrane. A signature of this asymmetry is the difference between the life times of adhesion bonds at front and rear, τ1 and τM, respectively, where τ1 ≫ τM. (b) There are four characteristic times for the membrane motion: The first time, T0 ˜ τM ˜ eaɛ, is the resting time where the displacement of the membrane is practically zero. The second time, Tp ˜ τ1 ˜ M, is the friction time which characterizes the time between two consecutive ruptures of adhesion bonds at the front, and which signalizes the onset of drift ("protrusion") at the leading edge. The third time, Tr ˜ Mγ(ɛ) (γ > 1), characterizes the "retraction" of the trailing edge, which is the retarded response to the pulling leading edge. The fourth time, TL ˜ M2, is the growth time for fluctuation of the end-to-end distance. (c) The separation of time scales, Tr/Tp ˜ Mγ(ɛ) - 1, leads to stretched fluctuations of the end-to-end distance, which are considered as stochastic cycles of protrusion and retraction on the time scale of TL. (d) The drift velocity v obeys anomalous scaling, {v} / p ˜ f(p^{1/γ (\\varepsilon )}M), where f (z) ˜ const. for small drag pM ≪ 1, and f (z) ˜ z-γ(ɛ) for pM ≫ 1, which implies {v} ˜ M^{-γ (\\varepsilon )}. These results may also turn out to be useful for the (more difficult

  14. Mitochondrial outer membrane forms bridge between two mitochondria in Arabidopsis thaliana.

    PubMed

    Yamashita, Akihiro; Fujimoto, Masaru; Katayama, Kenta; Tsutsumi, Nobuhiro; Arimura, Shin-Ichi

    2016-05-01

    Mitochondria are double-membrane organelles that move around and change their shapes dynamically. In plants, the dynamics of the outer membrane is not well understood. We recently demonstrated that mitochondria had tubular protrusions of the outer membrane with little or no matrix, called MOPs (mitochondrial outer-membrane protrusions; MOPs). Here we show that a MOP can form a bridge between two mitochondria in Arabidopsis thaliana. The bridge does not appear to involve the inner membranes. Live imaging revealed stretching of the MOP bridge, demonstrating the flexibility of the outer membrane. Mitochondria frequently undergo fission and fusion. These observations raise the possibility that MOPs bridges have a role in these processes. PMID:27031262

  15. The More the Tubular: Dynamic Bundling of Actin Filaments for Membrane Tube Formation

    PubMed Central

    Weichsel, Julian; Geissler, Phillip L.

    2016-01-01

    Tubular protrusions are a common feature of living cells, arising from polymerization of stiff protein filaments against a comparably soft membrane. Although this process involves many accessory proteins in cells, in vitro experiments indicate that similar tube-like structures can emerge without them, through spontaneous bundling of filaments mediated by the membrane. Using theory and simulation of physical models, we have elaborated how nonequilibrium fluctuations in growth kinetics and membrane shape can yield such protrusions. Enabled by a new grand canonical Monte Carlo method for membrane simulation, our work reveals a cascade of dynamical transitions from individually polymerizing filaments to highly cooperatively growing bundles as a dynamical bottleneck to tube formation. Filament network organization as well as adhesion points to the membrane, which bias filament bending and constrain membrane height fluctuations, screen the effective attractive interactions between filaments, significantly delaying bundling and tube formation. PMID:27384915

  16. Clostridium difficile toxin CDT induces formation of microtubule-based protrusions and increases adherence of bacteria.

    PubMed

    Schwan, Carsten; Stecher, Bärbel; Tzivelekidis, Tina; van Ham, Marco; Rohde, Manfred; Hardt, Wolf-Dietrich; Wehland, Jürgen; Aktories, Klaus

    2009-10-01

    Clostridium difficile causes antibiotic-associated diarrhea and pseudomembranous colitis by production of the Rho GTPase-glucosylating toxins A and B. Recently emerging hypervirulent Clostridium difficile strains additionally produce the binary ADP-ribosyltransferase toxin CDT (Clostridium difficile transferase), which ADP-ribosylates actin and inhibits actin polymerization. Thus far, the role of CDT as a virulence factor is not understood. Here we report by using time-lapse- and immunofluorescence microscopy that CDT and other binary actin-ADP-ribosylating toxins, including Clostridium botulinum C2 toxin and Clostridium perfringens iota toxin, induce redistribution of microtubules and formation of long (up to >150 microm) microtubule-based protrusions at the surface of intestinal epithelial cells. The toxins increase the length of decoration of microtubule plus-ends by EB1/3, CLIP-170 and CLIP-115 proteins and cause redistribution of the capture proteins CLASP2 and ACF7 from microtubules at the cell cortex into the cell interior. The CDT-induced microtubule protrusions form a dense meshwork at the cell surface, which wrap and embed bacterial cells, thereby largely increasing the adherence of Clostridia. The study describes a novel type of microtubule structure caused by less efficient microtubule capture and offers a new perspective for the pathogenetic role of CDT and other binary actin-ADP-ribosylating toxins in host-pathogen interactions.

  17. Fascin 2b Is a Component of Stereocilia that Lengthens Actin-Based Protrusions

    PubMed Central

    Gomez, Gustavo; Fernando, Carol A.; West, Megan C.; Pollock, Lana M.; Lin-Jones, Jennifer; Burnside, Beth; McDermott, Brian M.

    2011-01-01

    Stereocilia are actin-filled protrusions that permit mechanotransduction in the internal ear. To identify proteins that organize the cytoskeleton of stereocilia, we scrutinized the hair-cell transcriptome of zebrafish. One promising candidate encodes fascin 2b, a filamentous actin-bundling protein found in retinal photoreceptors. Immunolabeling of zebrafish hair cells and the use of transgenic zebrafish that expressed fascin 2b fused to green fluorescent protein demonstrated that fascin 2b localized to stereocilia specifically. When filamentous actin and recombinant fusion protein containing fascin 2b were combined in vitro to determine their dissociation constant, a Kd≈0.37 µM was observed. Electron microscopy showed that fascin 2b-actin filament complexes formed parallel actin bundles in vitro. We demonstrated that expression of fascin 2b or espin, another actin-bundling protein, in COS-7 cells induced the formation of long filopodia. Coexpression showed synergism between these proteins through the formation of extra-long protrusions. Using phosphomutant fascin 2b proteins, which mimicked either a phosphorylated or a nonphosphorylated state, in COS-7 cells and in transgenic hair cells, we showed that both formation of long filopodia and localization of fascin 2b to stereocilia were dependent on serine 38. Overexpression of wild-type fascin 2b in hair cells was correlated with increased stereociliary length relative to controls. These findings indicate that fascin 2b plays a key role in shaping stereocilia. PMID:21625653

  18. Keratocyte lamellipodial protrusion is characterized by a concave force-velocity relation.

    PubMed

    Heinemann, Fabian; Doschke, Holger; Radmacher, Manfred

    2011-03-16

    We report on the characterization of actin driven lamellipodial protrusion forces and velocities in keratocytes. A vertically mounted glass fiber acted as a flexible barrier positioned in front of migrating keratocytes with parallel phase contrast microscopy. A laser beam was coupled into the fiber and allowed detecting the position of the fiber by a segmented photodiode. Calibration of the fiber was carried out with the thermal oscillation method. Deflection and force signals were measured during lamellipodial protrusion. Velocity was constant during initial contact whereas loading force increased until finally the cell was stalled at higher forces. Stall forces were on the order of 2.9 ± 0.6 nN, which corresponds to a stall pressure of 2.7 ± 1.6 nN/μm(2). Assuming a density of actin filaments of 240 filaments per μm, we can estimate a stall force per actin filament of 1.7 ± 0.8 pN. To check for adaption of the cell against an external force, we let the cell push toward the glass fiber several times. On the timescale of the experiment (∼1 min), however, the cell did not adapt to previous loading events.

  19. Effect of tongue exercise on protrusive force and muscle fiber area in aging rats

    PubMed Central

    Connor, Nadine P.; Russell, John A.; Wang, Hao; Jackson, Michelle A.; Mann, Laura; Kluender, Keith R.

    2008-01-01

    Purpose Age-related changes in tongue function may contribute to dysphagia in elderly people. Our purpose was to investigate whether aged rats that have undergone tongue exercise would manifest increased protrusive tongue forces and increased genioglossus (GG) muscle fiber cross sectional areas. Method Forty-eight young adult, middle-aged and old Fischer 344/Brown Norway rats received 8 weeks of tongue exercise. Protrusive tongue forces were measured before and after exercise. GG muscle fiber cross sectional area was measured in exercised rats and compared with cross sectional areas in a no-exercise control group. Results A significant increase in maximum tongue force was found following exercise in all age groups. In addition, a trend for increased GG muscle fiber cross sectional area, and a significant increase in variability of GG muscle fiber cross sectional area were identified post-exercise. Conclusion The findings of this study have implications for treatment of elderly persons with dysphagia using tongue exercise programs. Specifically, increases in tongue force that occur following 8 weeks of progressive resistance tongue exercise may be accompanied by alterations in tongue muscle fiber morphology. These changes may provide greater strength and endurance for goal-oriented actions associated with the oropharyngeal swallow and should be investigated in future research. PMID:18723593

  20. Extraction of the lateral incisors to treat maxillary protrusion: quantitative evaluation of the stomatognathic functions.

    PubMed

    Ishihara, Yoshihito; Kuroda, Shingo; Sumiyoshi, Kumi; Takano-Yamamoto, Teruko; Yamashiro, Takashi

    2013-03-01

    To treat morphological abnormalities, impaction, and severe malposition of the teeth, the lateral incisors are sometimes extracted, followed by orthodontic space closure. This procedure often requires special consideration, not only with regard to esthetics but also for functional issues. However, thus far, few reports that have performed a functional evaluation in such cases. The purpose of this article is to report the successful treatment of an adult patient with a Class II division 1 malocclusion who was treated with extraction of the upper lateral incisors. The female patient, aged 23 years and 6 months, had a chief complaint of maxillary incisal protrusion and crooked teeth. In this patient, the upper lateral incisors were extracted to fulfill the patient's strong request, followed by orthodontic treatment using edgewise appliances. A high-pull J-hook headgear on the lower dental arch was used to prevent further labial inclination of the lower incisors. The total active treatment period was 37 months. The resulting occlusion and a satisfactory facial profile were maintained during a 4-year retention period. Additionally, this treatment did not affect the stomatognathic functions as assessed by the following criteria: range of the incisal path or condylar motion during maximal open-close movement, protrusive excursion, lateral excursion, and the chewing test. In conclusion, extraction of the upper lateral incisors can be an effective treatment choice when the upper lateral incisors are dwarfed, are nonvital, or demonstrate severe malposition.

  1. FMNL2 drives actin-based protrusion and migration downstream of Cdc42.

    PubMed

    Block, Jennifer; Breitsprecher, Dennis; Kühn, Sonja; Winterhoff, Moritz; Kage, Frieda; Geffers, Robert; Duwe, Patrick; Rohn, Jennifer L; Baum, Buzz; Brakebusch, Cord; Geyer, Matthias; Stradal, Theresia E B; Faix, Jan; Rottner, Klemens

    2012-06-01

    Cell migration entails protrusion of lamellipodia, densely packed networks of actin filaments at the cell front. Filaments are generated by nucleation, likely mediated by Arp2/3 complex and its activator Scar/WAVE. It is unclear whether formins contribute to lamellipodial actin filament nucleation or serve as elongators of filaments nucleated by Arp2/3 complex. Here we show that the Diaphanous-related formin FMNL2, also known as FRL3 or FHOD2, accumulates at lamellipodia and filopodia tips. FMNL2 is cotranslationally modified by myristoylation and regulated by interaction with the Rho-guanosine triphosphatase Cdc42. Abolition of myristoylation or Cdc42 binding interferes with proper FMNL2 activation, constituting an essential prerequisite for subcellular targeting. In vitro, C-terminal FMNL2 drives elongation rather than nucleation of actin filaments in the presence of profilin. In addition, filament ends generated by Arp2/3-mediated branching are captured and efficiently elongated by the formin. Consistent with these biochemical properties, RNAi-mediated silencing of FMNL2 expression decreases the rate of lamellipodia protrusion and, accordingly, the efficiency of cell migration. Our data establish that the FMNL subfamily member FMNL2 is a novel elongation factor of actin filaments that constitutes the first Cdc42 effector promoting cell migration and actin polymerization at the tips of lamellipodia. PMID:22608513

  2. New perspective on Herbst therapy for skeletal Class II malocclusions: a proposal for maxillary protrusion management.

    PubMed

    Filho, Leopoldino Capelozza; Siqueira, Danilo Furquim; de Castro, Renata Cristina Faria Ribeiro; An, Tien-Li; Cardoso, Mauricio de Almeida

    2012-01-01

    Angle Class II malocclusions may present morphologic deviations originated from the maxilla, mandible, or both. Since its reintroduction by Pancherz, the Herbst appliance has demonstrated effectiveness in the management of patients with mandibular deficiency. Because of the intermaxillary anchorage, the action of mandibular advancement provokes simultaneous reaction of maxillary restriction, similar to high-pull headgear. This aimed of this report is to compare two cases treated in two phases. In the first interceptive phase, the transverse problem was corrected by rapid maxillary expansion, which was followed by a Herbst appliance for mandibular advancement; in the second corrective phase, the cases were finished with fixed appliances. Although Herbst appliances were used in both patients, one patient with maxillary protrusion and another with mandibular deficiency, their use targeted different types of skeletal discrepancies. This difference allowed for the comparison of treatment effects, and although both patients had their malocclusion corrected, it seems reasonable to conclude that the final outcome was more favorable for the patient with maxillary protrusion.

  3. Multiple cytoskeletal pathways and PI3K signaling mediate CDC-42-induced neuronal protrusion in C. elegans.

    PubMed

    Alan, Jamie K; Struckhoff, Eric C; Lundquist, Erik A

    2013-01-01

    Rho GTPases are key regulators of cellular protrusion and are involved in many developmental events including axon guidance during nervous system development. Rho GTPase pathways display functional redundancy in developmental events, including axon guidance. Therefore, their roles can often be masked when using simple loss-of-function genetic approaches. As a complement to loss-of-function genetics, we constructed a constitutively activated CDC-42(G12V) expressed in C. elegans neurons. CDC-42(G12V) drove the formation of ectopic lamellipodial and filopodial protrusions in the PDE neurons, which resembled protrusions normally found on migrating growth cones of axons. We then used a candidate gene approach to identify molecules that mediate CDC-42(G12V)-induced ectopic protrusions by determining if loss of function of the genes could suppress CDC-42(G12V). Using this approach, we identified 3 cytoskeletal pathways previously implicated in axon guidance, the Arp2/3 complex, UNC-115/abLIM, and UNC-43/Ena. We also identified the Nck-interacting kinase MIG-15/NIK and p21-activated kinases (PAKs), also implicated in axon guidance. Finally, PI3K signaling was required, specifically the Rictor/mTORC2 branch but not the mTORC1 branch that has been implicated in other aspects of PI3K signaling including stress and aging. Our results indicate that multiple pathways can mediate CDC-42-induced neuronal protrusions that might be relevant to growth cone protrusions during axon pathfinding. Each of these pathways involves Rac GTPases, which might serve to integrate the pathways and coordinate the multiple CDC-42 pathways. These pathways might be relevant to developmental events such as axon pathfinding as well as disease states such as metastatic melanoma.

  4. Spontaneous resolution of a disc protrusion in a military aviator: a case report.

    PubMed

    Newlands, J C

    1997-04-01

    The case history of a 38-yr-old military aviator qualified on a range of rotary and fixed wing aircraft is presented. A radiologically evident and clinically debilitating (initially) lumbar disc protrusion was the source of the aviator's temporary grounding from flying duties. A full clinical recovery was made and he returned to flying, eventually to ejection seat aircraft. It is proposed that: 1.) wherever possible, initial conservative non-invasive management of back problems is preferable; 2.) lumbar disc lesions may sometimes totally spontaneously resolve (retraction or autolysis?), and; 3.) development and diagnosis of a lumbar disc lesion should not always be an automatic permanent expulsion from rotary wing or ejection seat aircraft. PMID:9096831

  5. Successful treatment of Class II malocclusion with bidental protrusion using standard edgewise prescription.

    PubMed

    Ayaz, Mohd; Kharbanda, Om Prakash

    2016-01-01

    This case report deals with the successful orthodontic treatment of a 14-year-old female patient having Class II malocclusion with bidental protrusion using standard edgewise prescription. She reported with forwardly placed upper front teeth and difficulty in closing lips. She had prognathic maxilla, retrognathic mandible, and full cusp Class II molar and canine relation bilaterally with overjet of 7 mm. She was in cervical vertebrae maturation indicator Stage IV. The case was treated by fixed extraction mechanotherapy. Interarch Class II mechanics was used to retract the upper incisor and to mesialize the lower molars. Simultaneously, Class I mechanics was used to upright lower incisors. Tip back bend, curve of Spee, and extra palatal root torque were incorporated in upper archwire to maintain molars in upright position and prevent extrusion and deepening of bite, respectively. There was satisfactory improvement in facial profile at the end of 24 months. After a follow-up of 6 months, occlusion was stable. PMID:27041906

  6. Successful treatment of Class II malocclusion with bidental protrusion using standard edgewise prescription

    PubMed Central

    Ayaz, Mohd; Kharbanda, Om Prakash

    2016-01-01

    This case report deals with the successful orthodontic treatment of a 14-year-old female patient having Class II malocclusion with bidental protrusion using standard edgewise prescription. She reported with forwardly placed upper front teeth and difficulty in closing lips. She had prognathic maxilla, retrognathic mandible, and full cusp Class II molar and canine relation bilaterally with overjet of 7 mm. She was in cervical vertebrae maturation indicator Stage IV. The case was treated by fixed extraction mechanotherapy. Interarch Class II mechanics was used to retract the upper incisor and to mesialize the lower molars. Simultaneously, Class I mechanics was used to upright lower incisors. Tip back bend, curve of Spee, and extra palatal root torque were incorporated in upper archwire to maintain molars in upright position and prevent extrusion and deepening of bite, respectively. There was satisfactory improvement in facial profile at the end of 24 months. After a follow-up of 6 months, occlusion was stable. PMID:27041906

  7. Severe bidentoalveolar protrusion treated with orthodontic microimplant-dependent en-masse retraction.

    PubMed

    Chung, Kyu-Rhim; Nelson, Gerald; Kim, Seong-Hun; Kook, Yoon-Ah

    2007-07-01

    This article describes the orthodontic treatment of a 14.5-year-old girl with severe bidentoalveolar protrusion. Specially designed sandblasted, large-grit, acid-etched (SLA) orthodontic microimplants (C-implants, Cimplant Co, Seoul, Korea) were placed in the alveolar bone in all 4 quadrants to provide anchorage for en-masse retraction without the help of banded or bonded molars. Successful retraction was achieved. The mandibular dentition was detailed by using conventional orthodontic appliances during the finishing stage. The osseointegration potential of these microimplants allows them to resist rotational force moments and control 3-dimensional movements of the anterior teeth during retraction. Facial esthetics improved for the patient, fullness of the upper and lower lips was reduced, and the interdental relationship was corrected. Biomechanical considerations, efficacy, and potential complications of the treatment technique are discussed.

  8. Penetrating keratoplasty for treatment of corneal protrusion in a great horned owl (Bubo virginianus).

    PubMed

    Andrew, Stacy E; Clippinger, Tracy L; Brooks, Dennis E; Helmick, Kelly E

    2002-09-01

    A young adult great horned owl (Bubo virginianus) was examined following presumed trauma. The owl had soft tissue injury to its left wing as well as corneal protrusion, lens subluxation, and iridodialysis of the right eye. The bird's eye was treated surgically with a large, rectangular penetrating keratoplasty. Following escape from housing, the bird was found with partial wound dehiscence and iris prolapse 12 days post operation. Surgical repair was performed and healing progressed for 14 days, at which time the transplant dehisced and the globe was exenterated. The patient rehabilitated well until escaping from its cage again 4 weeks later, at which time it sustained an open comminuted humeral fracture and was euthanized. PMID:12236872

  9. Cortico-muscular synchronization by proprioceptive afferents from the tongue muscles during isometric tongue protrusion.

    PubMed

    Maezawa, Hitoshi; Mima, Tatsuya; Yazawa, Shogo; Matsuhashi, Masao; Shiraishi, Hideaki; Funahashi, Makoto

    2016-03-01

    Tongue movements contribute to oral functions including swallowing, vocalizing, and breathing. Fine tongue movements are regulated through efferent and afferent connections between the cortex and tongue. It has been demonstrated that cortico-muscular coherence (CMC) is reflected at two frequency bands during isometric tongue protrusions: the beta (β) band at 15-35Hz and the low-frequency band at 2-10Hz. The CMC at the β band (β-CMC) reflects motor commands from the primary motor cortex (M1) to the tongue muscles through hypoglossal motoneuron pools. However, the generator mechanism of the CMC at the low-frequency band (low-CMC) remains unknown. Here, we evaluated the mechanism of low-CMC during isometric tongue protrusion using magnetoencephalography (MEG). Somatosensory evoked fields (SEFs) were also recorded following electrical tongue stimulation. Significant low-CMC and β-CMC were observed over both hemispheres for each side of the tongue. Time-domain analysis showed that the MEG signal followed the electromyography signal for low-CMC, which was contrary to the finding that the MEG signal preceded the electromyography signal for β-CMC. The mean conduction time from the tongue to the cortex was not significantly different between the low-CMC (mean, 80.9ms) and SEFs (mean, 71.1ms). The cortical sources of low-CMC were located significantly posterior (mean, 10.1mm) to the sources of β-CMC in M1, but were in the same area as tongue SEFs in the primary somatosensory cortex (S1). These results reveal that the low-CMC may be driven by proprioceptive afferents from the tongue muscles to S1, and that the oscillatory interaction was derived from each side of the tongue to both hemispheres. Oscillatory proprioceptive feedback from the tongue muscles may aid in the coordination of sophisticated tongue movements in humans.

  10. Effects of tongue-hold swallows on suprahyoid muscle activation according to the relative tongue protrusion length: a preliminary study.

    PubMed

    Oh, Jong-Chi

    2016-01-01

    Tongue-hold swallow (THS) is a therapeutic maneuver used to increase the posterior pharyngeal wall motion during swallowing. This maneuver has also been reported to result in increased activation of the suprahyoid muscles. The hypothesis of this study was that the degree of suprahyoid muscle activation would depend on the tongue protrusion-length. The aim of this study was to investigate the activation levels of the suprahyoid muscles by surface electromyography (sEMG) while performing the THS maneuver at three tongue-protrusion lengths. Suprahyoid muscle activity during THSs was recorded in 25 adult volunteers (17 women and 8 men; age range 20-38 years). To record the activity of the suprahyoid muscles while the participants performed the maneuver, surface wireless EMG electrodes separated by a distance of 1 cm were placed on the skin on both sides of the midline under the chin. Each activity was recorded three times. Data analysis was performed by repeated-measures analysis of variance. Our results revealed that participants exhibited greater electrical activity during THS with 2/3rd or maximal tongue protrusion as compared to THS with 1/3rd tongue protrusion (p ≤ 0.001). To maximize the therapeutic effect of the THS maneuver, it is advised to protrude the tongue maximally as long as swallowing is possible. PMID:27504242

  11. [Correction of a mandibular alveolar protrusion using maxillary miniscrew anchorage in the treatment of a Class II division 1 malocclusion].

    PubMed

    Haiim, Frederic

    2011-12-01

    This article describes a relatively simple method of reducing a mandibular alveolar protrusion without compensatory extraction of mandibular bicuspids or advancement of anterior teeth in the treatment of a Class II division 1 malocclusion. The use of well-planned skeletal anchorage specifically adapted to each specific situation is also well elucidated.

  12. Remotely Controlled Mandibular Protrusion during Sleep Predicts Therapeutic Success with Oral Appliances in Patients with Obstructive Sleep Apnea

    PubMed Central

    Remmers, John; Charkhandeh, Shouresh; Grosse, Joshua; Topor, Zbigniew; Brant, Rollin; Santosham, Peter; Bruehlmann, Sabina

    2013-01-01

    Study Objectives: The present study addresses the need for a validated tool that prospectively identifies favorable candidates for oral appliance therapy in treatment of obstructive sleep apnea. The objective of the study was to evaluate the ability of a mandibular titration study, performed with a remotely controlled mandibular positioner (RCMP), to predict treatment outcome with a mandibular repositioning appliance (MRA) and to predict an effective target protrusive position (ETPP). Design: A prospective, blinded, outcome study. Setting: Standard clinical care with tests performed in the polysomnographic laboratory. Participants: Consecutive patients (n = 67) recruited from a sleep center or a dental practice using broad inclusion criteria (age 21-80 years; AHI > 10/h; BMI < 40 kg/m2). Interventions: Therapeutic outcome with a mandibular protruding oral appliance was predicted following a mandibular protrusive titration study in the polysomnographic laboratory using a remotely controlled positioner and prospectively established predictive rules. An ETPP was also prospectively determined for participants predicted to be therapeutically successful with MRA therapy. All participants were blindly treated with a MRA, at either the predicted ETPP or a sham position, and therapeutic outcome was compared against prediction. Measurements and Results: At the final protrusive position, standard predictive parameters (sensitivity, specificity, positive and negative predictive values) showed statistically significant predictive accuracy (P < 0.05) in the range of 83% to 94%. The predicted ETPP provided an efficacious protrusive position in 87% of participants predicted to be therapeutically successful with MRA therapy (P < 0.05). Conclusions: Using prospectively established rules for interpreting the polysomnographic data, the mandibular titration study predicted mandibular repositioning appliance therapeutic outcome with significant accuracy, particularly with regard to

  13. Effects of mechanical tension on protrusive activity and microfilament and intermediate filament organization in an epidermal epithelium moving in culture

    PubMed Central

    1986-01-01

    Mechanical tension influences tissue morphogenesis and the synthetic, mitotic, and motile behavior of cells. To determine the effects of tension on epithelial motility and cytoskeletal organization, small, motile clusters of epidermal cells were artificially extended with a micromanipulated needle. Protrusive activity perpendicular to the axis of tension was dramatically suppressed. To determine the ultrastructural basis for this phenomenon, cells whose exact locomotive behavior was recorded cinemicrographically were examined by transmission electron microscopy. In untensed, forward-moving lamellar protrusions, microfilaments appear disorganized and anisotropically oriented. But in cytoplasm held under tension by micromanipulation or by the locomotive activity of other cells within the epithelium, microfilaments are aligned parallel to the tension. In non-spreading regions of the epithelial margin, microfilaments lie in tight bundles parallel to apparent lines of tension. Thus, it appears that tension causes alignment of microfilaments. In contrast, intermediate filaments are excluded from motile protrusions, being confined to the thicker, more central part of the cell. They roughly follow the contours of the cell, but are not aligned relative to tension even when microfilaments in the same cell are. This suggests that the organization of intermediate filaments is relatively resistant to physical distortion and the intermediate filaments may act as passive structural support within the cell. The alignment of microfilaments under tension suggests a mechanism by which tension suppresses protrusive activity: microfilaments aligned by forces exerted through filament-surface or filament-filament interconnections cannot reorient against such force and so cannot easily extend protrusions in directions not parallel to tension. PMID:3958054

  14. CLINICAL AND MAGNETIC RESONANCE IMAGING CHARACTERISTICS OF THORACOLUMBAR INTERVERTEBRAL DISK EXTRUSIONS AND PROTRUSIONS IN LARGE BREED DOGS.

    PubMed

    Gomes, Sergio A; Volk, Holger A; Packer, Rowena Ma; Kenny, Patrick J; Beltran, Elsa; De Decker, Steven

    2016-07-01

    Treatment recommendations differ for dogs with intervertebral disk extrusion vs. intervertebral disk protrusion. The aim of this retrospective, cross-sectional study was to determine whether clinical and magnetic resonance imaging (MRI) variables could be used to predict a diagnosis of thoracolumbar intervertebral disk extrusion or protrusion in dogs. Dogs were included if they were large breed dogs, had an MRI study of the thoracolumbar or lumbar vertebral column, had undergone spinal surgery, and had the type of intervertebral disk herniation (intervertebral disk extrusion or protrusion) clearly stated in surgical reports. A veterinary neurologist unaware of surgical findings reviewed MRI studies and recorded number, location, degree of degeneration and morphology of intervertebral disks, presence of nuclear clefts, disk space narrowing, extent, localization and lateralization of herniated disk material, degree of spinal cord compression, intraparenchymal intensity changes, spondylosis deformans, spinal cord swelling, spinal cord atrophy, vertebral endplate changes, and presence of extradural hemorrhage. Ninety-five dogs were included in the sample. Multivariable statistical models indicated that longer duration of clinical signs (P = 0.01), midline instead of lateralized disk herniation (P = 0.007), and partial instead of complete disk degeneration (P = 0.01) were associated with a diagnosis of intervertebral disk protrusion. The presence of a single intervertebral herniation (P = 0.023) and dispersed intervertebral disk material not confined to the disk space (P = 0.06) made a diagnosis of intervertebral disk extrusion more likely. Findings from this study identified one clinical and four MRI variables that could potentially facilitate differentiating intervertebral disk extrusions from protrusions in dogs. PMID:27038182

  15. The tail domain of myosin M catalyses nucleotide exchange on Rac1 GTPases and can induce actin-driven surface protrusions.

    PubMed

    Geissler, H; Ullmann, R; Soldati, T

    2000-05-01

    Members of the myosin superfamily play crucial roles in cellular processes including management of the cortical cytoskeleton, organelle transport and signal transduction. GTPases of the Rho family act as key control elements in the reorganization of the actin cytoskeleton in response to growth factors, and other functions such as membrane trafficking, transcriptional regulation, growth control and development. Here, we describe a novel unconventional myosin from Dictyostelium discoideum, MyoM. Primary sequence analysis revealed that it has the appearance of a natural chimera between a myosin motor domain and a guanine nucleotide exchange factor (GEF) domain for Rho GTPases. The functionality of both domains was established. Binding of the motor domain to F-actin was ATP-dependent and potentially regulated by phosphorylation. The GEF domain displayed selective activity on Rac1-related GTPases. Overexpression, rather than absence of MyoM, affected the cell morphology and viability. Particularly in response to hypo-osmotic stress, cells overexpressing the MyoM tail domain extended massive actin-driven protrusions. The GEF was enriched at the tip of growing protuberances, probably through its pleckstrin homology domain. MyoM is the first unconventional myosin containing an active Rac-GEF domain, suggesting a role at the interface of Rac-mediated signal transduction and remodeling of the actin cytoskeleton. PMID:11208126

  16. Supported lipid bilayer nanosystems: stabilization by undulatory-protrusion forces and destabilization by lipid bridging.

    PubMed

    Savarala, Sushma; Monson, Frederick; Ilies, Marc A; Wunder, Stephanie L

    2011-05-17

    Control of the stabilization/destabilization of supported lipid bilayers (SLBs) on nanoparticles is important for promotion of their organized assembly and for their use as delivery vehicles. At the same time, understanding the mechanism of these processes can yield insight into nanoparticle-cell interactions and nanoparticle toxicity. In this study, the suspension/precipitation process of zwitterionic lipid/SiO(2) nanosystems was analyzed as a function of ionic strength and as a function of the ratio of lipid/SiO(2) surface areas, at pH = 7.6. Salt is necessary to induce supported lipid bilayer (SLB) formation for zwitterionic lipids on silica (SiO(2)) (Seantier, B.; Kasemo, B., Influence of Mono- and Divalent Ions on the Formation of Supported Phospholipid Bilayers via Vesicle Adsorption. Langmuir 2009, 25 (10), 5767-5772). However, for zwitterionic SLBs on SiO(2) nanoparticles, addition of salt can cause precipitation of the SLBs, due to electrostatic shielding by both the lipid and the salt and to the suppression of thermal undulation/protrusion repulsive forces for lipids on solid surfaces. At ionic strengths that cause precipitation of SLBs, it was found that addition of excess SUVs, at ratios where there were equal populations of SUVs and SLBs, restored the undulation/protrusion repulsive forces and restabilized the suspensions. We suggest that SUVs separate SLBs in the suspension, as observed by TEM, and that SLB-SLB interactions are replaced by SLB-SUV interactions. Decreasing the relative amount of lipid, to the extent that there was less lipid available than the amount required for complete bilayer coverage of the SiO(2), resulted in precipitation of the nanosystem by a process of nanoparticle lipid bridging. For this case, we postulate a process in which lipid bilayer patches on one nanoparticle collide with bare silica patches on another SiO(2) nanoparticle, forming a single bilayer bridge between them. TEM data confirmed these findings, thus

  17. Plasmonic gratings with nano-protrusions made by glancing angle deposition for single-molecule super-resolution imaging

    NASA Astrophysics Data System (ADS)

    Chen, B.; Wood, A.; Pathak, A.; Mathai, J.; Bok, S.; Zheng, H.; Hamm, S.; Basuray, S.; Grant, S.; Gangopadhyay, K.; Cornish, P. V.; Gangopadhyay, S.

    2016-06-01

    Super-resolution imaging has been advantageous in studying biological and chemical systems, but the required equipment and platforms are expensive and unable to observe single-molecules at the high (μM) fluorophore concentrations required to study protein interaction and enzymatic activity. Here, a plasmonic platform was designed that utilized an inexpensively fabricated plasmonic grating in combination with a scalable glancing angle deposition (GLAD) technique using physical vapor deposition. The GLAD creates an abundance of plasmonic nano-protrusion probes that combine the surface plasmon resonance (SPR) from the periodic gratings with the localized SPR of these nano-protrusions. The resulting platform enables simultaneous imaging of a large area without point-by-point scanning or bulk averaging for the detection of single Cyanine-5 molecules in dye concentrations ranging from 50 pM to 10 μM using epifluorescence microscopy. Combining the near-field plasmonic nano-protrusion probes and super-resolution technique using localization microscopy, we demonstrate the ability to resolve grain sizes down to 65 nm. This plasmonic GLAD grating is a cost-effective super-resolution imaging substrate with potential applications in high-speed biomedical imaging over a wide range of fluorescent concentrations.Super-resolution imaging has been advantageous in studying biological and chemical systems, but the required equipment and platforms are expensive and unable to observe single-molecules at the high (μM) fluorophore concentrations required to study protein interaction and enzymatic activity. Here, a plasmonic platform was designed that utilized an inexpensively fabricated plasmonic grating in combination with a scalable glancing angle deposition (GLAD) technique using physical vapor deposition. The GLAD creates an abundance of plasmonic nano-protrusion probes that combine the surface plasmon resonance (SPR) from the periodic gratings with the localized SPR of these nano-protrusions

  18. Role of molecular tilt in thermal fluctuations of lipid membranes

    NASA Astrophysics Data System (ADS)

    May, Eric R.; Narang, Atul; Kopelevich, Dmitry I.

    2007-08-01

    Long-wavelength thermal fluctuations of lipid membranes are adequately described by the Helfrich elastic model. On the other hand, fluctuations of wavelengths comparable with bilayer thickness exhibit significant deviations from the prediction of the elastic model and are typically assumed to be dominated by microscopic surface tension due to protrusion of lipid molecules into the solvent. We present evidence that the short-wavelength modes of a lipid membrane are dominated by fluctuations of the tilt of lipid molecules with respect to the membrane normal rather than the microscopic surface tension. We obtain an expression for the spectral intensity of the thermal membrane fluctuations by appealing to the Hamm-Kozlov model, which accounts for both membrane bending and lipid tilt contributions to the total membrane energy but neglects the contributions of the microscopic surface tension. The tilt and the bending fluctuations obtained from our coarse-grained molecular dynamics simulations of a dipalmitoylphosphatidylcholine lipid bilayer show good agreement with the theory. Furthermore, the obtained tilt and bending moduli are in close agreement with experimentally determined values. The magnitude of the microscopic protrusion tension estimated from our simulations is significantly smaller than that of the tilt modulus. These results indicate that the membrane fluctuations can be adequately described by a macroscopic elastic model down to scales of interlipid distance provided one accounts for the tilt fluctuations.

  19. Robust Organizational Principles of Protrusive Biopolymer Networks in Migrating Living Cells

    PubMed Central

    Käs, Josef

    2011-01-01

    Cell migration is associated with the dynamic protrusion of a thin actin-based cytoskeletal extension at the cell front, which has been shown to consist of two different substructures, the leading lamellipodium and the subsequent lamellum. While the formation of the lamellipodium is increasingly well understood, organizational principles underlying the emergence of the lamellum are just beginning to be unraveled. We report here on a 1D mathematical model which describes the reaction-diffusion processes of a polarized actin network in steady state, and reproduces essential characteristics of the lamellipodium-lamellum system. We observe a steep gradient in filament lengths at the protruding edge, a local depolymerization maximum a few microns behind the edge, as well as a differential dominance of the network destabilizer ADF/cofilin and the stabilizer tropomyosin. We identify simple and robust organizational principles giving rise to the derived network characteristics, uncoupled from the specifics of any molecular implementation, and thus plausibly valid across cell types. An analysis of network length dependence on physico-chemical system parameters implies that to limit array treadmilling to cellular dimensions, network growth has to be truncated by mechanisms other than aging-induced depolymerization, e.g., by myosin-associated network dissociation at the transition to the cell body. Our work contributes to the analytical understanding of the cytoskeletal extension's bisection into lamellipodium and lamellum and sheds light on how cells organize their molecular machinery to achieve motility. PMID:21267070

  20. ER-endosome contact sites in endosome positioning and protrusion outgrowth.

    PubMed

    Raiborg, Camilla; Wenzel, Eva M; Pedersen, Nina M; Stenmark, Harald

    2016-04-15

    The endoplasmic reticulum (ER) makes abundant contacts with endosomes, and the numbers of contact sites increase as endosomes mature. It is already clear that such contact sites have diverse compositions and functions, but in this mini-review we will focus on two particular types of ER-endosome contact sites that regulate endosome positioning. Formation of ER-endosome contact sites that contain the cholesterol-binding protein oxysterol-binding protein-related protein 1L (ORP1L) is coordinated with loss of the minus-end-directed microtubule motor Dynein from endosomes. Conversely, formation of ER-endosome contact sites that contain the Kinesin-1-binding protein Protrudin results in transfer of the plus-end-directed microtubule motor Kinesin-1 from ER to endosomes. We discuss the possibility that formation of these two types of contact sites is coordinated as a 'gear-shift' mechanism for endosome motility, and we review evidence that Kinesin-1-mediated motility of late endosomes (LEs) to the cell periphery promotes outgrowth of neurites and other protrusions. PMID:27068952

  1. Exploratory Calibration of Adjustable-Protrusion Surface-Obstacle (APSO) Skin Friction Vector Gage

    NASA Technical Reports Server (NTRS)

    Hakkinen, Raimo J.; Neubauer, Jeremy S.; Hamory, Philip J.; Bui, Trong T.; Noffz, Gregory K.; Young, Ron (Technical Monitor)

    2003-01-01

    The design of an adjustable-protrusion surface-obstacle (APSO) skin friction vector gage is presented. Results from exploratory calibrations conducted in laminar and turbulent boundary layers at the Washington University Low-Speed Wind Tunnel and for turbulent boundary layers at speeds up to Mach 2 on the ceiling of the NASA Glenn Research Center 8- X 6-ft Supersonic Wind Tunnel are also discussed. The adjustable-height gage was designed to yield both the magnitude and direction of the surface shear stress vector and to measure the local static pressure distribution. Results from the NASA test show good correlation for subsonic and low supersonic conditions covering several orders of magnitude in terms of the adopted similarity variables. Recommendations for future work in this area consist of identifying the physical parameters responsible for the disagreement between the university and NASA data sets, developing a compressibility correction specific to the APSO geometry, and examining the effect that static pressure distribution and skewed boundary layers have on the results from the APSO.

  2. Biomechanical implications of excessive endograft protrusion into the aortic arch after thoracic endovascular repair.

    PubMed

    Rinaudo, Antonino; Raffa, Giuseppe Maria; Scardulla, Francesco; Pilato, Michele; Scardulla, Cesare; Pasta, Salvatore

    2015-11-01

    Endografts placed in the aorta for thoracic endovascular aortic repair (TEVAR) may determine malappositioning to the lesser curvature of the aortic wall, thus resulting in a devastating complication known as endograft collapse. This premature device failure commonly occurs in young individuals after TEVAR for traumatic aortic injuries as a result of applications outside the physical conditions for which the endograft was designed. In this study, an experimentally-calibrated fluid-structure interaction (FSI) model was developed to assess the hemodynamic and stress/strain distributions acting on the excessive protrusion extension (PE) of endografts deployed in four young patients underwent TEVAR. Endograft infolding was experimentally measured for different hemodynamic scenarios by perfusion testing and then used to numerically calibrate the mechanical behavior of endograft PE. Results evinced that the extent of endograft can severely alter the hemodynamic and structural loads exerted on the endograft PE. Specifically, PE determined a physiological aortic coarctation into the aortic arch characterized by a helical flow in the distal descending aorta. High device displacement and transmural pressure across the stent-graft wall were found for a PE longer than 21 mm. Finally, marked intramural stress and principal strain distributions on the protruded segment of the endograft wall may suggest failure due to material fatigue. These critical parameters may contribute to the endograft collapse observed clinically and can be used to design new devices more suitable for young individuals to be treated with an endoprosthesis for TEVAR of blunt traumatic aortic injuries.

  3. Spontaneous Regression of Herniated Lumbar Disc with New Disc Protrusion in the Adjacent Level

    PubMed Central

    Gürcan, Serkan

    2016-01-01

    Spontaneous regression of herniated lumbar discs was reported occasionally. The mechanisms proposed for regression of disc herniation are still incomplete. This paper describes and discusses a case of spontaneous regression of herniated lumbar discs with a new disc protrusion in the adjacent level. A 41-year-old man was admitted with radiating pain and numbness in the left lower extremity with a left posterolateral disc extrusion at L5-S1 level. He was admitted to hospital with low back pain due to disc herniation caudally immigrating at L4-5 level three years ago. He refused the surgical intervention that was offered and was treated conservatively at that time. He had no neurological deficit and a history of spontaneous regression of the extruded lumbar disc; so, a conservative therapy, including bed rest, physical therapy, nonsteroidal anti-inflammatory drugs, and analgesics, was advised. In conclusion, herniated lumbar disc fragments may regress spontaneously. Reports are prone to advise conservative treatment for extruded or sequestrated lumbar disc herniations. However, these patients should be followed up closely; new herniation at adjacent/different level may occur. Furthermore, it is important to know which herniated disk should be removed and which should be treated conservatively, because disc herniation may cause serious complications as muscle weakness and cauda equine syndrome. PMID:27429818

  4. Spontaneous Regression of Herniated Lumbar Disc with New Disc Protrusion in the Adjacent Level.

    PubMed

    Hakan, Tayfun; Gürcan, Serkan

    2016-01-01

    Spontaneous regression of herniated lumbar discs was reported occasionally. The mechanisms proposed for regression of disc herniation are still incomplete. This paper describes and discusses a case of spontaneous regression of herniated lumbar discs with a new disc protrusion in the adjacent level. A 41-year-old man was admitted with radiating pain and numbness in the left lower extremity with a left posterolateral disc extrusion at L5-S1 level. He was admitted to hospital with low back pain due to disc herniation caudally immigrating at L4-5 level three years ago. He refused the surgical intervention that was offered and was treated conservatively at that time. He had no neurological deficit and a history of spontaneous regression of the extruded lumbar disc; so, a conservative therapy, including bed rest, physical therapy, nonsteroidal anti-inflammatory drugs, and analgesics, was advised. In conclusion, herniated lumbar disc fragments may regress spontaneously. Reports are prone to advise conservative treatment for extruded or sequestrated lumbar disc herniations. However, these patients should be followed up closely; new herniation at adjacent/different level may occur. Furthermore, it is important to know which herniated disk should be removed and which should be treated conservatively, because disc herniation may cause serious complications as muscle weakness and cauda equine syndrome. PMID:27429818

  5. Trans-anal protrusion of ventriculo-peritoneal shunt catheter with silent bowel perforation: report of ten cases in children.

    PubMed

    Ghritlaharey, Rajendra Kumar; Budhwani, K S; Shrivastava, Dhirendra K; Gupta, Gaurav; Kushwaha, Anand Singh; Chanchlani, Roshan; Nanda, Monika

    2007-06-01

    Ventriculo-peritoneal (VP) shunting used in the treatment for hydrocephalus is associated with several complications. Mechanical failure of shunt is the commonest complication of all. Visceral/bowel perforation is an unusual but serious complication of VP shunting. This article reports our experience in the management of ten children who had VP Shunt catheter protrusion from anus. This is a retrospective study of ten patients who had VP shunt catheter protrusion from anus, admitted in the department of paediatric surgery between Jan 1996 and Dec 2005. The records of above ten cases were reviewed for their clinical presentation and management, etc. We had performed 398 VP shunt operations in the last 10 years. Two hundred and seventy one (68.09%) VP Shunts were done for congenital hydrocephalus of which 164 were done in infancy/neonatal period and 107 VP shunts were done in the age group of >1-12 years. One hundred and twenty-seven (31.90%) VP shunt operations were done for patients who had hydrocephalus as a complication following tubercular meningitis (TBM). Out of 398 VP shunts, ten patients (2.51%) had protrusion of the distal end of peritoneal catheter from anus without causing/leading to peritonitis. We observed a 08.29% mortality of all VP shunt operations. Protrusion of VP shunt catheter per rectum can occur without producing peritonitis. Formal exploration and localization of entry of VP shunt catheter in bowel is not mandatory. Mini laparotomy and revision of peritoneal part of shunt can be done if there is no shunt infection.

  6. Plasmonic gratings with nano-protrusions made by glancing angle deposition for single-molecule super-resolution imaging.

    PubMed

    Chen, B; Wood, A; Pathak, A; Mathai, J; Bok, S; Zheng, H; Hamm, S; Basuray, S; Grant, S; Gangopadhyay, K; Cornish, P V; Gangopadhyay, S

    2016-06-16

    Super-resolution imaging has been advantageous in studying biological and chemical systems, but the required equipment and platforms are expensive and unable to observe single-molecules at the high (μM) fluorophore concentrations required to study protein interaction and enzymatic activity. Here, a plasmonic platform was designed that utilized an inexpensively fabricated plasmonic grating in combination with a scalable glancing angle deposition (GLAD) technique using physical vapor deposition. The GLAD creates an abundance of plasmonic nano-protrusion probes that combine the surface plasmon resonance (SPR) from the periodic gratings with the localized SPR of these nano-protrusions. The resulting platform enables simultaneous imaging of a large area without point-by-point scanning or bulk averaging for the detection of single Cyanine-5 molecules in dye concentrations ranging from 50 pM to 10 μM using epifluorescence microscopy. Combining the near-field plasmonic nano-protrusion probes and super-resolution technique using localization microscopy, we demonstrate the ability to resolve grain sizes down to 65 nm. This plasmonic GLAD grating is a cost-effective super-resolution imaging substrate with potential applications in high-speed biomedical imaging over a wide range of fluorescent concentrations.

  7. Three-dimensional matrix fiber alignment modulates cell migration and MT1-MMP utility by spatially and temporally directing protrusions

    PubMed Central

    Fraley, Stephanie I.; Wu, Pei-hsun; He, Lijuan; Feng, Yunfeng; Krisnamurthy, Ranjini; Longmore, Gregory D.; Wirtz, Denis

    2015-01-01

    Multiple attributes of the three-dimensional (3D) extracellular matrix (ECM) have been independently implicated as regulators of cell motility, including pore size, crosslink density, structural organization, and stiffness. However, these parameters cannot be independently varied within a complex 3D ECM protein network. We present an integrated, quantitative study of these parameters across a broad range of complex matrix configurations using self-assembling 3D collagen and show how each parameter relates to the others and to cell motility. Increasing collagen density resulted in a decrease and then an increase in both pore size and fiber alignment, which both correlated significantly with cell motility but not bulk matrix stiffness within the range tested. However, using the crosslinking enzyme Transglutaminase II to alter microstructure independently of density revealed that motility is most significantly predicted by fiber alignment. Cellular protrusion rate, protrusion orientation, speed of migration, and invasion distance showed coupled biphasic responses to increasing collagen density not predicted by 2D models or by stiffness, but instead by fiber alignment. The requirement of matrix metalloproteinase (MMP) activity was also observed to depend on microstructure, and a threshold of MMP utility was identified. Our results suggest that fiber topography guides protrusions and thereby MMP activity and motility. PMID:26423227

  8. Three-dimensional matrix fiber alignment modulates cell migration and MT1-MMP utility by spatially and temporally directing protrusions.

    PubMed

    Fraley, Stephanie I; Wu, Pei-Hsun; He, Lijuan; Feng, Yunfeng; Krisnamurthy, Ranjini; Longmore, Gregory D; Wirtz, Denis

    2015-01-01

    Multiple attributes of the three-dimensional (3D) extracellular matrix (ECM) have been independently implicated as regulators of cell motility, including pore size, crosslink density, structural organization, and stiffness. However, these parameters cannot be independently varied within a complex 3D ECM protein network. We present an integrated, quantitative study of these parameters across a broad range of complex matrix configurations using self-assembling 3D collagen and show how each parameter relates to the others and to cell motility. Increasing collagen density resulted in a decrease and then an increase in both pore size and fiber alignment, which both correlated significantly with cell motility but not bulk matrix stiffness within the range tested. However, using the crosslinking enzyme Transglutaminase II to alter microstructure independently of density revealed that motility is most significantly predicted by fiber alignment. Cellular protrusion rate, protrusion orientation, speed of migration, and invasion distance showed coupled biphasic responses to increasing collagen density not predicted by 2D models or by stiffness, but instead by fiber alignment. The requirement of matrix metalloproteinase (MMP) activity was also observed to depend on microstructure, and a threshold of MMP utility was identified. Our results suggest that fiber topography guides protrusions and thereby MMP activity and motility. PMID:26423227

  9. Three-dimensional matrix fiber alignment modulates cell migration and MT1-MMP utility by spatially and temporally directing protrusions

    NASA Astrophysics Data System (ADS)

    Fraley, Stephanie I.; Wu, Pei-Hsun; He, Lijuan; Feng, Yunfeng; Krisnamurthy, Ranjini; Longmore, Gregory D.; Wirtz, Denis

    2015-10-01

    Multiple attributes of the three-dimensional (3D) extracellular matrix (ECM) have been independently implicated as regulators of cell motility, including pore size, crosslink density, structural organization, and stiffness. However, these parameters cannot be independently varied within a complex 3D ECM protein network. We present an integrated, quantitative study of these parameters across a broad range of complex matrix configurations using self-assembling 3D collagen and show how each parameter relates to the others and to cell motility. Increasing collagen density resulted in a decrease and then an increase in both pore size and fiber alignment, which both correlated significantly with cell motility but not bulk matrix stiffness within the range tested. However, using the crosslinking enzyme Transglutaminase II to alter microstructure independently of density revealed that motility is most significantly predicted by fiber alignment. Cellular protrusion rate, protrusion orientation, speed of migration, and invasion distance showed coupled biphasic responses to increasing collagen density not predicted by 2D models or by stiffness, but instead by fiber alignment. The requirement of matrix metalloproteinase (MMP) activity was also observed to depend on microstructure, and a threshold of MMP utility was identified. Our results suggest that fiber topography guides protrusions and thereby MMP activity and motility.

  10. Cdc42 and Actin Control Polarized Expression of TI-VAMP Vesicles to Neuronal Growth Cones and Their Fusion with the Plasma MembraneV⃞

    PubMed Central

    Alberts, Philipp; Rudge, Rachel; Irinopoulou, Theano; Danglot, Lydia; Gauthier-Rouvière, Cécile; Galli, Thierry

    2006-01-01

    Tetanus neurotoxin-insensitive vesicle-associated membrane protein (TI-VAMP)-mediated fusion of intracellular vesicles with the plasma membrane is crucial for neurite outgrowth, a pathway not requiring synaptobrevin-dependent exocytosis. Yet, it is not known how the TI-VAMP membrane trafficking pathway is regulated or how it is coordinated with cytoskeletal dynamics within the growth cone that guide neurite outgrowth. Here, we demonstrate that TI-VAMP, but not synaptobrevin 2, concentrates in the peripheral, F-actin-rich region of the growth cones of hippocampal neurons in primary culture. Its accumulation correlates with and depends upon the presence of F-actin. Moreover, acute stimulation of actin remodeling by homophilic activation of the adhesion molecule L1 induces a site-directed, actin-dependent recruitment of the TI-VAMP compartment. Expression of a dominant-positive mutant of Cdc42, a key regulator of cell polarity, stimulates formation of F-actin- and TI-VAMP-rich filopodia outside the growth cone. Furthermore, we report that Cdc42 activates exocytosis of pHLuorin tagged TI-VAMP in an actin-dependent manner. Collectively, our data suggest that Cdc42 and regulated assembly of the F-actin network control the accumulation and exocytosis of TI-VAMP-containing membrane vesicles in growth cones to coordinate membrane trafficking and actin remodeling during neurite outgrowth. PMID:16381811

  11. Cdc42 and actin control polarized expression of TI-VAMP vesicles to neuronal growth cones and their fusion with the plasma membrane.

    PubMed

    Alberts, Philipp; Rudge, Rachel; Irinopoulou, Theano; Danglot, Lydia; Gauthier-Rouvière, Cécile; Galli, Thierry

    2006-03-01

    Tetanus neurotoxin-insensitive vesicle-associated membrane protein (TI-VAMP)-mediated fusion of intracellular vesicles with the plasma membrane is crucial for neurite outgrowth, a pathway not requiring synaptobrevin-dependent exocytosis. Yet, it is not known how the TI-VAMP membrane trafficking pathway is regulated or how it is coordinated with cytoskeletal dynamics within the growth cone that guide neurite outgrowth. Here, we demonstrate that TI-VAMP, but not synaptobrevin 2, concentrates in the peripheral, F-actin-rich region of the growth cones of hippocampal neurons in primary culture. Its accumulation correlates with and depends upon the presence of F-actin. Moreover, acute stimulation of actin remodeling by homophilic activation of the adhesion molecule L1 induces a site-directed, actin-dependent recruitment of the TI-VAMP compartment. Expression of a dominant-positive mutant of Cdc42, a key regulator of cell polarity, stimulates formation of F-actin- and TI-VAMP-rich filopodia outside the growth cone. Furthermore, we report that Cdc42 activates exocytosis of pHLuorin tagged TI-VAMP in an actin-dependent manner. Collectively, our data suggest that Cdc42 and regulated assembly of the F-actin network control the accumulation and exocytosis of TI-VAMP-containing membrane vesicles in growth cones to coordinate membrane trafficking and actin remodeling during neurite outgrowth.

  12. Distinct Roles for the N- and C-terminal Regions of M-Sec in Plasma Membrane Deformation during Tunneling Nanotube Formation.

    PubMed

    Kimura, Shunsuke; Yamashita, Masami; Yamakami-Kimura, Megumi; Sato, Yusuke; Yamagata, Atsushi; Kobashigawa, Yoshihiro; Inagaki, Fuyuhiko; Amada, Takako; Hase, Koji; Iwanaga, Toshihiko; Ohno, Hiroshi; Fukai, Shuya

    2016-01-01

    The tunneling nanotube (TNT) is a structure used for intercellular communication, and is a thin membrane protrusion mediating transport of various signaling molecules and cellular components. M-Sec has potent membrane deformation ability and induces TNT formation in cooperation with the Ral/exocyst complex. Here, we show that the N-terminal polybasic region of M-Sec directly binds phosphatidylinositol (4,5)-bisphosphate for its localization to the plasma membrane during the initial stage of TNT formation. We further report a crystal structure of M-Sec, which consists of helix bundles arranged in a straight rod-like shape, similar to the membrane tethering complex subunits. A positively charged surface in the C-terminal domains is required for M-Sec interaction with active RalA to extend the plasma membrane protrusions. Our results suggest that the membrane-associated M-Sec recruits active RalA, which directs the exocyst complex to form TNTs.

  13. Targeted Disruption of Shp2 in Chondrocytes Leads to Metachondromatosis With Multiple Cartilaginous Protrusions

    PubMed Central

    Kim, Harry KW; Feng, Gen-Sheng; Chen, Di; King, Philip D; Kamiya, Nobuhiro

    2014-01-01

    Metachondromatosis is a benign bone disease predominantly observed in the hands and feet of children or young adults demonstrating two different manifestations: a cartilage-capped bony outgrowth on the surface of the bone called exostosis and ectopic cartilaginous nodules inside the bone called enchondroma. Recently, it has been reported that loss-of-function mutations of the SHP2 gene, which encodes the SHP2 protein tyrosine phosphatase, are associated with metachondromatosis. The purpose of this study was to investigate the role of SHP2 in postnatal cartilage development, which is largely unknown. We disrupted Shp2 during the postnatal stage of mouse development in a chondrocyte-specific manner using a tamoxifen-inducible system. We found tumor-like nodules on the hands and feet within a month after the initial induction. The SHP2-deficient mice demonstrated an exostosis-like and enchondroma-like phenotype in multiple bones of the hands, feet, and ribs as assessed by X-ray and micro-computed tomography (CT). Histological assessment revealed the disorganization of the growth plate cartilage, a cartilaginous protrusion from the epiphyseal bone, and ectopic cartilage nodules within the bones, which is consistent with the pathological features of metachondromatosis in humans (ie, both exostosis and enchondroma). At molecular levels, we observed an abundant expression of Indian hedgehog protein (IHH) and fibroblast growth factor 2 (FGF2) and impaired expression of mitogen-activated protein kinases (MAPK) in the affected cartilage nodules in the SHP2-deficient mice. In summary, we have generated a mouse model of metachondromatosis that includes manifestations of exostosis and enchondroma. This study provides a novel model for the investigation of the pathophysiology of the disease and advances the understanding of metachondromatosis. This model will be useful to identify molecular mechanisms for the disease cause and progression as well as to develop new therapeutic

  14. Urinary Continence after Robot-Assisted Laparoscopic Radical Prostatectomy: The Impact of Intravesical Prostatic Protrusion

    PubMed Central

    Jo, Jung Ki; Hong, Sung Kyu; Byun, Seok-Soo; Zargar, Homayoun; Autorino, Riccardo

    2016-01-01

    Purpose To assess the impact of intravesical prostatic protrusion (IPP) on the outcomes of robot-assisted laparoscopic prostatectomy (RALP). Materials and Methods The medical records of 1094 men who underwent RALP from January 2007 to March 2013 were analyzed using our database to identify 641 additional men without IPP (non-IPP group). We excluded 259 patients who presented insufficient data and 14 patients who did not have an MRI image. We compared the following parameters: preoperative transrectal ultrasound, prostate specific antigen (PSA), clinicopathologic characteristics, intraoperative characteristics, postoperative oncologic characteristics, minor and major postoperative complications, and continence until postoperative 1 year. IPP grade was stratified by grade into three groups: Grade 1 (IPP≤5 mm), Grade 2 (5 mm10 mm). Results Of the 821 patients who underwent RALP, 557 (67.8%) experienced continence at postoperative 3 months, 681 (82.9%) at 6 months, and 757 (92.2%) at 12 months. According to IPP grade, there were significant differences in recovering full continence at postoperative 3 months, 6 months, and 12 months (p<0.001). On multivariate analysis, IPP was the most powerful predictor of postoperative continence in patients who underwent RALP (p<0.001). Using a generalized estimating equation model, IPP also was shown to be the most powerful independent variable for postoperative continence in patients who underwent RALP (p<0.001). Conclusion Patients with low-grade IPP have significantly higher chances of recovering full continence. Therefore, the known IPP grade will be helpful during consultations with patients before RALP. PMID:27401645

  15. EB1-recruited microtubule +TIP complexes coordinate protrusion dynamics during 3D epithelial remodeling

    PubMed Central

    Gierke, Sarah; Wittmann, Torsten

    2012-01-01

    SUMMARY Background Epithelial remodeling, in which apical-basal polarized cells switch to a migratory phenotype, plays a central role in development and disease of multicellular organisms. Although dynamic microtubules (MTs) are required for directed migration on flat surfaces, how MT dynamics are controlled or contribute to epithelial remodeling in a more physiological three-dimensional (3D) environment is not understood. We use confocal live cell imaging to analyze MT function and dynamics during 3D epithelial morphogenesis and remodeling of polarized Madin-Darby canine kidney (MDCK) epithelial cells that undergo partial epithelial-to-mesenchymal transition (EMT) in response to hepatocyte growth factor (HGF). Results We find that HGF treatment increases MT growth rate before morphological changes are evident, and that large numbers of MTs grow into HGF-induced cell extensions independent of centrosome reorientation. Using lentivirus-mediated shRNA, we demonstrate that EB1, an adaptor protein that mediates recruitment of numerous other +TIP proteins to growing MT plus ends, is required for this HGF-induced MT reorganization. We further show that protrusion and adhesion dynamics are disorganized, and that vesicular trafficking to the tip of HGF-induced cell extensions is disrupted in EB1-depleted cells. Conclusions We conclude that EB1-mediated interactions with growing MTs are important to coordinate cell shape changes and directed migration into the surrounding extracellular matrix during epithelial remodeling in a physiological 3D environment. In contrast, EB1 is not required for the establishment or maintenance of apical-basal cell polarity, suggesting different functions of +TIPs and MTs in different types of cell polarity. PMID:22483942

  16. Comparison of Mallampati test with lower jaw protrusion maneuver in predicting difficult laryngoscopy and intubation

    PubMed Central

    Ul Haq, Muhammad Irfan; Ullah, Hameed

    2013-01-01

    Background: Failure to maintain a patent airway is one of the commonest causes of anesthesia-related morbidity and mortality. Many protocols, algorithms, and different combinations of tested methods for airway assessment have been developed to predict difficult laryngoscopy and intubation. The reported incidence of a difficult intubation varies from 1.5% to 13%. The objective of this study was to compare Mallampati test (MT) with lower jaw protrusion (LJP) maneuver in predicting difficult laryngoscopy and intubation. Materials and Methods: Seven hundred and sixty patients were included in the study. All the patients underwent MT and LJP maneuver for their airway assessment. After a standardized technique of induction of anesthesia, primary anesthetist performed laryngoscopy and graded it according to the grades described by Cormack and Lehane. Sensitivity, specificity, accuracy, and positive predictive value (PPV) and negative predictive value (NPV) were calculated for both these tests with 95% confidence interval (CI) using conventional laryngoscopy as gold standard. Area under curve was also calculated for both, MT and LJP maneuver. A P < 0.05 was taken as significant. Results: LJP maneuver had higher sensitivity (95.9% vs. 27.1%), NPV (98.7% vs. 82.0%), and accuracy (90.1% vs. 80.3%) when compared to MT in predicting difficult laryngoscopy and intubation. Both tests, however, had similar specificity and PPV. There was marked difference in the positive and negative likelihood ratio between LJP and MT. Similarly, the area under the curve favored LJP maneuver over MT. Conclusion: The results of this study show that LJP maneuver is a better test to predict difficult laryngoscopy and tracheal intubation. We recommend the addition of this maneuver to the routine preoperative evaluation of airway. PMID:24106353

  17. Membrane Tension Acts Through PLD2 and mTORC2 to Limit Actin Network Assembly During Neutrophil Migration

    PubMed Central

    Diz-Muñoz, Alba; Thurley, Kevin; Chintamen, Sana; Altschuler, Steven J.; Fletcher, Daniel A.; Weiner, Orion D.

    2016-01-01

    For efficient polarity and migration, cells need to regulate the magnitude and spatial distribution of actin assembly. This process is coordinated by reciprocal interactions between the actin cytoskeleton and mechanical forces. Actin polymerization-based protrusion increases tension in the plasma membrane, which in turn acts as a long-range inhibitor of actin assembly. These interactions form a negative feedback circuit that limits the magnitude of membrane tension in neutrophils and prevents expansion of the existing front and the formation of secondary fronts. It has been suggested that the plasma membrane directly inhibits actin assembly by serving as a physical barrier that opposes protrusion. Here we show that efficient control of actin polymerization-based protrusion requires an additional mechanosensory feedback cascade that indirectly links membrane tension with actin assembly. Specifically, elevated membrane tension acts through phospholipase D2 (PLD2) and the mammalian target of rapamycin complex 2 (mTORC2) to limit actin nucleation. In the absence of this pathway, neutrophils exhibit larger leading edges, higher membrane tension, and profoundly defective chemotaxis. Mathematical modeling suggests roles for both the direct (mechanical) and indirect (biochemical via PLD2 and mTORC2) feedback loops in organizing cell polarity and motility—the indirect loop is better suited to enable competition between fronts, whereas the direct loop helps spatially organize actin nucleation for efficient leading edge formation and cell movement. This circuit is essential for polarity, motility, and the control of membrane tension. PMID:27280401

  18. Membrane Tension Acts Through PLD2 and mTORC2 to Limit Actin Network Assembly During Neutrophil Migration.

    PubMed

    Diz-Muñoz, Alba; Thurley, Kevin; Chintamen, Sana; Altschuler, Steven J; Wu, Lani F; Fletcher, Daniel A; Weiner, Orion D

    2016-06-01

    For efficient polarity and migration, cells need to regulate the magnitude and spatial distribution of actin assembly. This process is coordinated by reciprocal interactions between the actin cytoskeleton and mechanical forces. Actin polymerization-based protrusion increases tension in the plasma membrane, which in turn acts as a long-range inhibitor of actin assembly. These interactions form a negative feedback circuit that limits the magnitude of membrane tension in neutrophils and prevents expansion of the existing front and the formation of secondary fronts. It has been suggested that the plasma membrane directly inhibits actin assembly by serving as a physical barrier that opposes protrusion. Here we show that efficient control of actin polymerization-based protrusion requires an additional mechanosensory feedback cascade that indirectly links membrane tension with actin assembly. Specifically, elevated membrane tension acts through phospholipase D2 (PLD2) and the mammalian target of rapamycin complex 2 (mTORC2) to limit actin nucleation. In the absence of this pathway, neutrophils exhibit larger leading edges, higher membrane tension, and profoundly defective chemotaxis. Mathematical modeling suggests roles for both the direct (mechanical) and indirect (biochemical via PLD2 and mTORC2) feedback loops in organizing cell polarity and motility-the indirect loop is better suited to enable competition between fronts, whereas the direct loop helps spatially organize actin nucleation for efficient leading edge formation and cell movement. This circuit is essential for polarity, motility, and the control of membrane tension.

  19. A novel mechanism of generating extracellular vesicles during apoptosis via a beads-on-a-string membrane structure.

    PubMed

    Atkin-Smith, Georgia K; Tixeira, Rochelle; Paone, Stephanie; Mathivanan, Suresh; Collins, Christine; Liem, Michael; Goodall, Katharine J; Ravichandran, Kodi S; Hulett, Mark D; Poon, Ivan K H

    2015-06-15

    Disassembly of apoptotic cells into smaller fragments (a form of extracellular vesicle called apoptotic bodies) can facilitate removal of apoptotic debris and intercellular communication. However, the mechanism underpinning this process is unclear. While observing monocytes undergoing apoptosis by time-lapse microscopy, we discovered a new type of membrane protrusion that resembles a 'beads-on-a-string' structure. Strikingly, the 'beads' are frequently sheared off the 'string' to form apoptotic bodies. Generation of apoptotic bodies via this mechanism can facilitate a sorting process and results in the exclusion of nuclear contents from apoptotic bodies. Mechanistically, generation of 'beads-on-a-string' protrusion is controlled by the level of actomyosin contraction and apoptopodia formation. Furthermore, in an unbiased drug screen, we identified the ability of sertraline (an antidepressant) to block the formation of 'beads-on-a-string' protrusions and apoptotic bodies. These data uncover a new mechanism of apoptotic body formation in monocytes and also compounds that can modulate this process.

  20. Infantile Perianal Pyramidal Protrusion with Coexisting Perineal and Perianal Hemangiomas: A Fortuitous Association or Incomplete PELVIS Syndrome?

    PubMed Central

    Verma, Shyam B; Wollina, Uwe

    2014-01-01

    Two cases of infantile perianal pyramidal protrusions (IPPP), one pyramidal in shape and one leaf shaped, are being described by us. Both were notable for coexisting hemangiomas in the close vicinity. To the best of our knowledge we are reporting these two variants of IPPP with the associated neighboring hemangiomas for the first time in Indian dermatologic literature. We suggest that this presentation may be a part of one of the syndromes that comprise anorectal malformations with hemangiomas like PELVIS syndrome and others mentioned in the table. PMID:24470664

  1. Bovine binder-of-sperm protein BSP1 promotes protrusion and nanotube formation from liposomes

    SciTech Connect

    Lafleur, Michel; Courtemanche, Lesley; Karlsson, Goeran; Edwards, Katarina; Schwartz, Jean-Louis; Manjunath, Puttaswamy

    2010-08-27

    Research highlights: {yields} Binder-of-sperm protein 1 (BSP1) modifies the morphology of lipidic vesicles inducing bead necklace-like and thread-like structures. {yields} In the presence of multilamellar liposomes, BSP1 leads to the formation of long nanotubes. {yields} The insertion of BSP1 in the external lipid leaflet of membranes induces local changes in bilayer curvature. -- Abstract: Binder-of-sperm (BSP) proteins interact with sperm membranes and are proposed to extract selectively phosphatidylcholine and cholesterol from these. This change in lipid composition is a key step in sperm capacitation. The present work demonstrates that the interactions between the protein BSP1 and model membranes composed with phosphatidylcholine lead to drastic changes in the morphology of the lipidic self-assemblies. Using cryo-electron microscopy and fluorescence microscopy, we show that, in the presence of the protein, the lipid vesicles elongate, and form bead necklace-like structures that evolve toward small vesicles or thread-like structures. In the presence of multilamellar vesicles, where a large reservoir of lipid is available, the presence of BSP proteins lead to the formation of long nanotubes. Long spiral-like threads, associated with lipid/protein complexes, are also observed. The local curvature of lipid membranes induced by the BSP proteins may be involved in lipid domain formation and the extraction of some lipids during the sperm maturation process.

  2. Membrane stabilizer

    DOEpatents

    Mingenbach, William A.

    1988-01-01

    A device is provided for stabilizing a flexible membrane secured within a frame, wherein a plurality of elongated arms are disposed radially from a central hub which penetrates the membrane, said arms imposing alternately against opposite sides of the membrane, thus warping and tensioning the membrane into a condition of improved stability. The membrane may be an opaque or translucent sheet or other material.

  3. Examination of unsteady flow in a mildly curved vessel with stent-like wall protrusions: A tale of two vessels

    NASA Astrophysics Data System (ADS)

    Prince, Chekema; Peterson, Sean D.

    2014-11-01

    New stent designs allow for better conformity to the vessel curvature, maintaining the complex primary and secondary flow patterns present in the native vessel. Despite design improvements, stent induced alterations in local vascular geometry are inevitable and have been associated with stent failure due to in-stent restenosis (ISR). The objective of this study is to elucidate the unsteady flow physics induced by stent implantation, accounting in particular for vessel curvature. The present study focuses on the investigation of unsteady flow through mildly curved vessels with protrusion patterns that emulate current stent designs using computational fluid dynamics (CFD). The modeled geometries include various protrusion frequencies, heights, and widths. Two different arterial velocities waveforms, mimicking the coronary and carotid artery environment, will be considered. A detailed examination of the flow environment induced by the stent presence will be correlated with derived parameters from the flow behavior, such as critical wall shear stress typically associated with ISR development. Specifically, the role of secondary flow in the convective transport of ISR stimuli to the vessel wall will be explored.

  4. Regulation of membrane-shape transitions induced by I-BAR domains.

    PubMed

    Chen, Zhiming; Shi, Zheng; Baumgart, Tobias

    2015-07-21

    I-BAR proteins are well-known actin-cytoskeleton adaptors and have been observed to be involved in the formation of plasma membrane protrusions (filopodia). I-BAR proteins contain an all-helical, crescent-shaped IRSp53-MIM domain (IMD) dimer that is believed to be able to couple with a membrane shape. This coupling could involve the sensing and even the generation of negative plasma membrane curvature. Indeed, the in vitro studies have shown that IMDs can induce inward tubulation of liposomes. While N-BAR domains, which generate positive membrane curvature, have received a considerable amount of attention from both theory and experiments, the mechanisms of curvature coupling through IMDs are comparatively less studied and understood. Here we used a membrane-shape stability assay developed recently in our lab to quantitatively characterize IMD-induced membrane-shape transitions. We determined a membrane-shape stability diagram for IMDs that reveals how membrane tension and protein density can comodulate the generation of IMD-induced membrane protrusions. From comparison to analytical theory, we determine three key parameters that characterize the curvature coupling of IMD. We find that the curvature generation capacity of IMDs is significantly stronger compared to that of endophilin, an N-BAR protein known to be involved in plasma membrane shape transitions. Contrary to N-BAR domains, where amphipathic helix insertion is known to promote its membrane curvature generation, for IMDs we find that amphipathic helices inhibit membrane shape transitions, consistent with the inverse curvature that IMDs generate. Importantly, in both of these types of BAR domains, electrostatic interactions affect membrane-binding capacity, but do not appear to affect the curvature generation capacity of the protein. These two types of BAR domain proteins show qualitatively similar membrane shape stability diagrams, suggesting an underlying ubiquitous mechanism by which peripheral proteins

  5. Processivity and collectivity of biomolecular motors extracting membrane nanotubes.

    PubMed

    Fontenele Araujo, Francisco; Storm, Cornelis

    2012-07-01

    Biomolecular motors can pull and viscously drag membranes. The resulting elongations include cell protrusions, tether networks, and sensorial tentacles. Here we focus on the extraction of a single tube from a vesicle. Via a force balance coupled to binding kinetics, we analytically determine the phase diagram of tube formation as function of the motor processivity, the surface viscosity of the membrane η'(m), and the density of motors on the vesicle ρ. Three tubulation mechanisms are identified: (i) tip pulling, due to the accumulation of motors at the leading edge of the membrane, (ii) viscous drag, emergent from the translation of motors along the tube, and (iii) hybrid extraction, such that tip pulling and viscous drag are equally important. For experimental values of η'(m) and ρ, we find that the growth of bionanotubes tends to be driven by viscous forces, whereas artificial membranes are dominated by tip pulling.

  6. Dynamic recruitment of the curvature-sensitive protein ArhGAP44 to nanoscale membrane deformations limits exploratory filopodia initiation in neurons

    PubMed Central

    Galic, Milos; Tsai, Feng-Chiao; Collins, Sean R; Matis, Maja; Bandara, Samuel; Meyer, Tobias

    2014-01-01

    In the vertebrate central nervous system, exploratory filopodia transiently form on dendritic branches to sample the neuronal environment and initiate new trans-neuronal contacts. While much is known about the molecules that control filopodia extension and subsequent maturation into functional synapses, the mechanisms that regulate initiation of these dynamic, actin-rich structures have remained elusive. Here, we find that filopodia initiation is suppressed by recruitment of ArhGAP44 to actin-patches that seed filopodia. Recruitment is mediated by binding of a membrane curvature-sensing ArhGAP44 N-BAR domain to plasma membrane sections that were deformed inward by acto-myosin mediated contractile forces. A GAP domain in ArhGAP44 triggers local Rac-GTP hydrolysis, thus reducing actin polymerization required for filopodia formation. Additionally, ArhGAP44 expression increases during neuronal development, concurrent with a decrease in the rate of filopodia formation. Together, our data reveals a local auto-regulatory mechanism that limits initiation of filopodia via protein recruitment to nanoscale membrane deformations. DOI: http://dx.doi.org/10.7554/eLife.03116.001 PMID:25498153

  7. Cytosolic pressure provides a propulsive force comparable to actin polymerization during lamellipod protrusion

    PubMed Central

    Manoussaki, Daphne; Shin, William D.; Waterman, Clare M.; Chadwick, Richard S.

    2015-01-01

    Does cytosolic pressure facilitate f-actin polymerization to push the leading edge of a cell forward during self-propelled motion? AFM force-distance (f-d) curves obtained from lamellipodia of live cells often exhibit a signal from which the tension, bending modulus, elastic modulus and thickness in the membrane-cortex complex can be estimated close to the contact point. These measurements permit an estimate of the cytosolic pressure via the canonical Laplace force balance. The deeper portion of the f-d curve allows estimation of the bulk modulus of the cytoskeleton after removal of the bottom effect artifact. These estimates of tension, pressure, cortex thickness and elastic moduli imply that cytosolic pressure both pushes the membrane forward and compresses the actin cortex rearward to facilitate f-actin polymerization. We also estimate that cytosolic pressure fluctuations, most likely induced by myosin, provide a propulsive force comparable to that provided by f-actin polymerization in a lamellipod. PMID:26197304

  8. Membrane stabilizer

    DOEpatents

    Mingenbach, W.A.

    1988-02-09

    A device is provided for stabilizing a flexible membrane secured within a frame, wherein a plurality of elongated arms are disposed radially from a central hub which penetrates the membrane, said arms imposing alternately against opposite sides of the membrane, thus warping and tensioning the membrane into a condition of improved stability. The membrane may be an opaque or translucent sheet or other material. 10 figs.

  9. A novel approach to pseudopodia proteomics: excimer laser etching, two-dimensional difference gel electrophoresis, and confocal imaging

    PubMed Central

    Mimae, Takahiro; Ito, Akihiko; Hagiyama, Man; Nakanishi, Jun; Hosokawa, Yoichiroh; Okada, Morihito; Murakami, Yoshinori; Kondo, Tadashi

    2014-01-01

    Pseudopodia are actin-rich ventral cellular protrusions shown to facilitate the migration and metastasis of tumor cells. Here, we present a novel approach to perform pseudopodia proteomics. Tumor cells growing on porous membranes extend pseudopodia into the membrane pores. In our method, cell bodies are removed by horizontal ablation at the basal cell surface with the excimer laser while pseudopodia are left in the membrane pores. For protein expression profiling, whole cell and pseudopodia proteins are extracted with a lysis buffer, labeled with highly sensitive fluorescent dyes, and separated by two-dimensional gel electrophoresis. Proteins with unique expression patterns in pseudopodia are identified by mass spectrometry. The effects of the identified proteins on pseudopodia formation are evaluated by measuring the pseudopodia length in cancer cells with genetically modified expression of target proteins using confocal imaging. This protocol allows global identification of pseudopodia proteins and evaluation of their functional significance in pseudopodia formation within one month. PMID:25309719

  10. Membrane Elastic Properties and Cell Function

    PubMed Central

    Pontes, Bruno; Ayala, Yareni; Fonseca, Anna Carolina C.; Romão, Luciana F.; Amaral, Racκele F.; Salgado, Leonardo T.; Lima, Flavia R.; Farina, Marcos; Viana, Nathan B.; Moura-Neto, Vivaldo; Nussenzveig, H. Moysés

    2013-01-01

    Recent studies indicate that the cell membrane, interacting with its attached cytoskeleton, is an important regulator of cell function, exerting and responding to forces. We investigate this relationship by looking for connections between cell membrane elastic properties, especially surface tension and bending modulus, and cell function. Those properties are measured by pulling tethers from the cell membrane with optical tweezers. Their values are determined for all major cell types of the central nervous system, as well as for macrophage. Astrocytes and glioblastoma cells, which are considerably more dynamic than neurons, have substantially larger surface tensions. Resting microglia, which continually scan their environment through motility and protrusions, have the highest elastic constants, with values similar to those for resting macrophage. For both microglia and macrophage, we find a sharp softening of bending modulus between their resting and activated forms, which is very advantageous for their acquisition of phagocytic functions upon activation. We also determine the elastic constants of pure cell membrane, with no attached cytoskeleton. For all cell types, the presence of F-actin within tethers, contrary to conventional wisdom, is confirmed. Our findings suggest the existence of a close connection between membrane elastic constants and cell function. PMID:23844071

  11. Neural tube closure in Xenopus laevis involves medial migration, directed protrusive activity, cell intercalation and convergent extension.

    PubMed

    Davidson, L A; Keller, R E

    1999-10-01

    We have characterized the cell movements and prospective cell identities as neural folds fuse during neural tube formation in Xenopus laevis. A newly developed whole-mount, two-color fluorescent RNA in situ hybridization method, visualized with confocal microscopy, shows that the dorsal neural tube gene xpax3 and the neural-crest-specific gene xslug are expressed far lateral to the medial site of neural fold fusion and that expression moves medially after fusion. To determine whether cell movements or dynamic changes in gene expression are responsible, we used low-light videomicroscopy followed by fluorescent in situ and confocal microscopy. These methods revealed that populations of prospective neural crest and dorsal neural tube cells near the lateral margin of the neural plate at the start of neurulation move to the dorsal midline using distinctive forms of motility. Before fold fusion, superficial neural cells apically contract, roll the neural plate into a trough and appear to pull the superficial epidermal cell sheet medially. After neural fold fusion, lateral deep neural cells move medially by radially intercalating between other neural cells using two types of motility. The neural crest cells migrate as individual cells toward the dorsal midline using medially directed monopolar protrusions. These movements combine the two lateral populations of neural crest into a single medial population that form the roof of the neural tube. The remaining cells of the dorsal neural tube extend protrusions both medially and laterally bringing about radial intercalation of deep and superficial cells to form a single-cell-layered, pseudostratified neural tube. While ours is the first description of medially directed cell migration during neural fold fusion and re-establishment of the neural tube, these complex cell behaviors may be involved during cavitation of the zebrafish neural keel and secondary neurulation in the posterior axis of chicken and mouse.

  12. Pinkbar is an epithelial-specific BAR domain protein that generates planar membrane structures

    SciTech Connect

    Pykäläinen, Anette; Boczkowska, Malgorzata; Zhao, Hongxia; Saarikangas, Juha; Rebowski, Grzegorz; Jansen, Maurice; Hakanen, Janne; Koskela, Essi V.; Peränen, Johan; Vihinen, Helena; Jokitalo, Eija; Salminen, Marjo; Ikonen, Elina; Dominguez, Roberto; Lappalainen, Pekka

    2013-05-29

    Bin/amphipysin/Rvs (BAR)-domain proteins sculpt cellular membranes and have key roles in processes such as endocytosis, cell motility and morphogenesis. BAR domains are divided into three subfamilies: BAR- and F-BAR-domain proteins generate positive membrane curvature and stabilize cellular invaginations, whereas I-BAR-domain proteins induce negative curvature and stabilize protrusions. We show that a previously uncharacterized member of the I-BAR subfamily, Pinkbar, is specifically expressed in intestinal epithelial cells, where it localizes to Rab13-positive vesicles and to the plasma membrane at intercellular junctions. Notably, the BAR domain of Pinkbar does not induce membrane tubulation but promotes the formation of planar membrane sheets. Structural and mutagenesis analyses reveal that the BAR domain of Pinkbar has a relatively flat lipid-binding interface and that it assembles into sheet-like oligomers in crystals and in solution, which may explain its unique membrane-deforming activity.

  13. Membrane Supply and Demand Regulates F-Actin in a Cell Surface Reservoir.

    PubMed

    Figard, Lauren; Wang, Mengyu; Zheng, Liuliu; Golding, Ido; Sokac, Anna Marie

    2016-05-01

    Cells store membrane in surface reservoirs of pits and protrusions. These membrane reservoirs facilitate cell shape change and buffer mechanical stress, but we do not know how reservoir dynamics are regulated. During cellularization, the first cytokinesis in Drosophila embryos, a reservoir of microvilli unfolds to fuel cleavage furrow ingression. We find that regulated exocytosis adds membrane to the reservoir before and during unfolding. Dynamic F-actin deforms exocytosed membrane into microvilli. Single microvilli extend and retract in ∼20 s, while the overall reservoir is depleted in sync with furrow ingression over 60-70 min. Using pharmacological and genetic perturbations, we show that exocytosis promotes microvillar F-actin assembly, while furrow ingression controls microvillar F-actin disassembly. Thus, reservoir F-actin and, consequently, reservoir dynamics are regulated by membrane supply from exocytosis and membrane demand from furrow ingression.

  14. Quantitative Three-Dimensional Ultrasound Analysis of Tongue Protrusion, Grooving and Symmetry: Data from 12 Normal Speakers and a Partial Glossectomee

    ERIC Educational Resources Information Center

    Bressmann, Tim; Thind, Parveen; Uy, Catherine; Bollig, Carmen; Gilbert, Ralph W.; Irish, Jonathan C.

    2005-01-01

    The functional determinants for a good speech outcome after a partial tongue resection and reconstruction are not well established. The purpose of the present study was to assess the protrusion, grooving and symmetry of the tongue during sustained speech sound production using three-dimensional ultrasound. The participants were twelve normal…

  15. Automatic Prompting and Positive Attention to Reduce Tongue Protrusion and Head Tilting by Two Adults with Severe to Profound Intellectual Disabilities

    ERIC Educational Resources Information Center

    Lancioni, Giulio E.; Singh, Nirbhay N.; O'Reilly, Mark F.; Sigafoos, Jeff; Didden, Robert; Pichierri, Sabrina

    2010-01-01

    This study assessed a simple behavioral strategy for reducing stereotypic tongue protrusion and forward head tilting displayed by a woman and a man with severe to profound intellectual disabilities. The strategy involved (a) auditory prompting (i.e., verbal encouragements to keep the tongue in the mouth or the head upright) delivered automatically…

  16. Evaluation of condylar inclination of dentulous subjects determined by axiograph and to compare with manual programming of articulators using protrusive interocclusal record

    PubMed Central

    Prasad, Krishna D.; Shetty, Manoj; Chandy, Binoj K.

    2015-01-01

    Aims: To determine the average condylar inclination using ultrasonic axiograph; to determine the average condylar inclination using protrusive interocclusal bite records; to compare whether there is any marked difference in the values obtained by these techniques. Settings and Design: This clinical study compares the mean horizontal condylar inclination of the ultrasonic axiograph (Axioquick system) and the manual programming using protrusive interocclusal records. Materials and Methods: The study was conducted on a group of 30 subjects reporting to Department of Prosthodontics. Axioquick software system of SAM III (School Articulator Munich) fully-adjustable articulator was used, and interocclusal recording of condylar inclination is the manual method used in semi-adjustable articulators using Aluwax. The condylar inclination of the articulator was adjusted and set using protrusive interocclusal record. Statistical Analysis: The horizontal condylar inclination values of both methods were collected, and the mean of right and left condylar inclination was compared using paired t-test. Results: A statistically significant difference exists between Axioquick system and the manual method of programming articulator with protrusive interocclusal records (P ≤ 0.001). Conclusions: Within the limitations of the present study, the following conclusions were drawn: The average condylar inclination by axiograph is 42.125°. The average condylar inclination by interocclusal record is 33.25°. Comparison of both values shows a difference of 8.88° ± 4.03° that showed a significant difference (P < 0.001). PMID:26321837

  17. Molecular modeling of membrane responses to the adsorption of rotating nanoparticles: promoted cell uptake and mechanical membrane rupture.

    PubMed

    Yue, Tongtao; Zhang, Xianren; Huang, Fang

    2015-01-21

    Recently, a unique dynamic magnetic field was developed to induce the rotational movement of superparamagnetic iron oxide nanoparticles. This technique has been applied to remotely control both cellular internalization and apoptosis. Therefore, a thorough understanding of how a lipid membrane responds to the introduction of rotating NPs is quite important to promote the applications of this technique in a variety of biomedical area. Here, we performed Dissipative Particle Dynamics (DPD) simulations to systematically investigate the interaction mechanism between lipid membranes and rotating NPs. Two kinds of membrane responses are observed. One is the promoted cell uptake and the other is the mechanical membrane rupture. The promoting effect of NP rotation on the cell uptake is ascribed to the enhanced membrane monolayer protrusion, which can wrap the NP from the top side. Meanwhile, the rotating NP exerts a shearing force on the membrane. Accordingly, the membrane undergoes a local distortion around the NP. If the shearing force exceeds a critical value, the local membrane distortion develops into a mechanical rupture. A number of factors, like NP size, NP shape, ligand density and rotation speed, are critical in both of the above membrane responses. PMID:25388826

  18. Membrane tension and membrane fusion.

    PubMed

    Kozlov, Michael M; Chernomordik, Leonid V

    2015-08-01

    Diverse cell biological processes that involve shaping and remodeling of cell membranes are regulated by membrane lateral tension. Here we focus on the role of tension in driving membrane fusion. We discuss the physics of membrane tension, forces that can generate the tension in plasma membrane of a cell, and the hypothesis that tension powers expansion of membrane fusion pores in late stages of cell-to-cell and exocytotic fusion. We propose that fusion pore expansion can require unusually large membrane tensions or, alternatively, low line tensions of the pore resulting from accumulation in the pore rim of membrane-bending proteins. Increase of the inter-membrane distance facilitates the reaction. PMID:26282924

  19. Electrostatic and electrokinetic contributions to the elastic moduli of a driven membrane

    NASA Astrophysics Data System (ADS)

    Lacoste, D.; Menon, G. I.; Bazant, M. Z.; Joanny, J. F.

    2009-03-01

    We discuss the electrostatic contribution to the elastic moduli of a cell or artificial membrane placed in an electrolyte and driven by a DC electric field. The field drives ion currents across the membrane, through specific channels, pumps or natural pores. In steady state, charges accumulate in the Debye layers close to the membrane, modifying the membrane elastic moduli. We first study a model of a membrane of zero thickness, later generalizing this treatment to allow for a finite thickness and finite dielectric constant. Our results clarify and extend the results presented by D. Lacoste, M. Cosentino Lagomarsino, and J.F. Joanny (EPL 77, 18006 (2007)), by providing a physical explanation for a destabilizing term proportional to k ⊥ 3 in the fluctuation spectrum, which we relate to a nonlinear (E2) electrokinetic effect called induced-charge electro-osmosis (ICEO). Recent studies of ICEO have focused on electrodes and polarizable particles, where an applied bulk field is perturbed by capacitive charging of the double layer and drives the flow along the field axis toward surface protrusions; in contrast, we predict “reverse” ICEO flows around driven membranes, due to curvature-induced tangential fields within a nonequilibrium double layer, which hydrodynamically enhance protrusions. We also consider the effect of incorporating the dynamics of a spatially dependent concentration field for the ion channels.

  20. Membrane dynamics at the nuclear exchange junction during early mating (one to four hours) in the ciliate Tetrahymena thermophila.

    PubMed

    Cole, Eric S; Giddings, Thomas H; Ozzello, Courtney; Winey, Mark; O'Toole, Eileen; Orias, Judy; Hamilton, Eileen; Guerrier, Sabrice; Ballard, Anna; Aronstein, Tyler

    2015-02-01

    Using serial-section transmission electron microscopy and three-dimensional (3D) electron tomography, we characterized membrane dynamics that accompany the construction of a nuclear exchange junction between mating cells in the ciliate Tetrahymena thermophila. Our methods revealed a number of previously unknown features. (i) Membrane fusion is initiated by the extension of hundreds of 50-nm-diameter protrusions from the plasma membrane. These protrusions extend from both mating cells across the intercellular space to fuse with membrane of the mating partner. (ii) During this process, small membrane-bound vesicles or tubules are shed from the plasma membrane and into the extracellular space within the junction. The resultant vesicle-filled pockets within the extracellular space are referred to as junction lumens. (iii) As junction lumens fill with extracellular microvesicles and swell, the plasma membrane limiting these swellings undergoes another deformation, pinching off vesicle-filled vacuoles into the cytoplasm (reclamation). (iv) These structures (resembling multivesicular bodies) seem to associate with autophagosomes abundant near the exchange junction. We propose a model characterizing the membrane-remodeling events that establish cytoplasmic continuity between mating Tetrahymena cells. We also discuss the possible role of nonvesicular lipid transport in conditioning the exchange junction lipid environment. Finally, we raise the possibility of an intercellular signaling mechanism involving microvesicle shedding and uptake.

  1. Membrane Dynamics at the Nuclear Exchange Junction during Early Mating (One to Four Hours) in the Ciliate Tetrahymena thermophila

    PubMed Central

    Giddings, Thomas H.; Ozzello, Courtney; Winey, Mark; O'Toole, Eileen; Orias, Judy; Hamilton, Eileen; Guerrier, Sabrice; Ballard, Anna; Aronstein, Tyler

    2014-01-01

    Using serial-section transmission electron microscopy and three-dimensional (3D) electron tomography, we characterized membrane dynamics that accompany the construction of a nuclear exchange junction between mating cells in the ciliate Tetrahymena thermophila. Our methods revealed a number of previously unknown features. (i) Membrane fusion is initiated by the extension of hundreds of 50-nm-diameter protrusions from the plasma membrane. These protrusions extend from both mating cells across the intercellular space to fuse with membrane of the mating partner. (ii) During this process, small membrane-bound vesicles or tubules are shed from the plasma membrane and into the extracellular space within the junction. The resultant vesicle-filled pockets within the extracellular space are referred to as junction lumens. (iii) As junction lumens fill with extracellular microvesicles and swell, the plasma membrane limiting these swellings undergoes another deformation, pinching off vesicle-filled vacuoles into the cytoplasm (reclamation). (iv) These structures (resembling multivesicular bodies) seem to associate with autophagosomes abundant near the exchange junction. We propose a model characterizing the membrane-remodeling events that establish cytoplasmic continuity between mating Tetrahymena cells. We also discuss the possible role of nonvesicular lipid transport in conditioning the exchange junction lipid environment. Finally, we raise the possibility of an intercellular signaling mechanism involving microvesicle shedding and uptake. PMID:25107923

  2. Higher-order assemblies of BAR domain proteins for shaping membranes.

    PubMed

    Suetsugu, Shiro

    2016-06-01

    Most cellular organelles contain lipid bilayer membranes. The earliest characterization of cellular organelles was performed by electron microscopy observation of such membranes. However, the precise mechanisms for shaping the membrane in particular subcellular organelles is poorly understood. Classically, the overall cellular shape, i.e. the shape of the plasma membrane, was thought to be governed by the reorganization of cytoskeletal components such as actin and microtubules. The plasma membrane contains various submicron structures such as clathrin-coated pits, caveolae, filopodia and lamellipodia. These subcellular structures are either invaginations or protrusions and are associated with the cytoskeleton. Therefore, it could be hypothesized that there are membrane-binding proteins that cooperates with cytoskeleton in shaping of plasma membrane organelles. Proteins with the Bin-Amphiphysin-Rvs (BAR) domain connect a variety of membrane shapes to actin filaments. The BAR domains themselves bend the membranes by their rigidity and then mold the membranes into tubules through their assembly as spiral polymers, which are thought to be involved in the various submicron structures. Membrane tubulation by polymeric assembly of the BAR domains is supposed to be regulated by binding proteins, binding lipids and the mechanical properties of the membrane. This review gives an overview of BAR protein assembly, describes the significance of the assembly and discusses how to study the assembly in the context of membrane and cellular morphology. The technical problems encountered in microscopic observation of BAR domain assembly are also discussed.

  3. Membrane distillation

    NASA Astrophysics Data System (ADS)

    Bryk, Mikhail T.; Nigmatullin, R. R.

    1994-12-01

    Studies in the field of membrane distillation are analysed. A critical analysis of the theoretical and experimental investigations of membrane distillation is presented. Attention is concentrated on the mechanism of mass transfer and the influence of various external factors on the process characteristics. Questions concerning the creation of modules and apparatus for membrane distillation and aspects of the practical employment of such distillation in order to obtain pure water, for the purification of waste water, and for the concentration of technological solutions in various branches of industry are considered quite fully. The advantages and disadvantages of membrane distillation compared with other membrane methods are analysed. The bibliography includes 97 references.

  4. Dementia with Lewy bodies presenting marked tongue protrusion and bite due to lingual dystonia: A case report.

    PubMed

    Shiga, Yuji; Kanaya, Yuhei; Kono, Ryuhei; Takeshima, Shinichi; Shimoe, Yutaka; Kuriyama, Masaru

    2016-06-22

    We report the patient of a 53-year-old woman who developed subacute-onset marked tonge protrusion and bite. She was diagnosed as dementia with Lewy bodies (DLB) from the clinical features including progressive cognitive decline, visual hallucinations, parkinsonism, and severe insomnia and depression, and the radiological finding of low dopamine transported uptake in basal ganglia by Dat SCAN and low blood circulation in occipital lobe of cerebrum. The patient received 600 mg doses of levodopa for over a year, followed by rotigotine and ropinirole with a rapid increase of dosage. It is believed that these treatments stimulated and sensitized dopamine D1 receptors, thereby inducing lingual dystonia. Furthermore, the patient demonstrated dyspnea and attacks of apnea caused by the closure of bilateral vocal cords due to laryngeal dyskinesia. After initiation of the neuroleptic, olanzapine, for a short duration, the high dose of levodopa overlapped with neuroleptic sensitivity, suggesting DOPA-induced dystonia and dyskinesia. This interaction can sometimes lead to lethal adverse events, and must be considered very important when treating patients with DLB. PMID:27212676

  5. Bimaxillary protrusion with an atrophic alveolar defect: orthodontics, autogenous chin-block graft, soft tissue augmentation, and an implant.

    PubMed

    Chiu, Grace S C; Chang, Chris H N; Roberts, W Eugene

    2015-01-01

    Bimaxillary protrusion in a 28-year-old woman was complicated by multiple missing, restoratively compromised, or hopeless teeth. The maxillary right central incisor had a history of avulsion and replantation that subsequently evolved into generalized external root resorption with Class III mobility and severe loss of the supporting periodontium. This complex malocclusion had a discrepancy index of 21, and 8 additional points were scored for the atrophic dental implant site (maxillary right central incisor). The comprehensive treatment plan included extraction of 4 teeth (both maxillary first premolars, the maxillary right central incisor, and the mandibular right first molar), orthodontic closure of all spaces except for the future implant site (maxillary right central incisor), augmentation of the alveolar defect with an autogenous chin-block graft, enhancement of the gingival biotype with a connective tissue graft, and an implant-supported prosthesis. Orthodontists must understand the limitations of bone grafts. Augmented alveolar defects are slow to completely turn over to living bone, so they are usually good sites for implants but respond poorly to orthodontic space closure. However, postsurgical orthodontic treatment is often indicated to optimally finish the esthetic zone before placing the final prosthesis. The latter was effectively performed for this patient, resulting in a total treatment time of about 36 months for comprehensive interdisciplinary care. An excellent functional and esthetic result was achieved. PMID:25533077

  6. Rubisco decrease is involved in chloroplast protrusion and Rubisco-containing body formation in soybean (Glycine max.) under salt stress.

    PubMed

    He, Yi; Yu, Chenliang; Zhou, Li; Chen, Yue; Liu, Ao; Jin, Junhua; Hong, Jian; Qi, Yanhua; Jiang, Dean

    2014-01-01

    Salt stress often induces declination of net photosynthetic rate (Pn), partially resulted from Rubisco degradation. The chloroplast protrusions (CPs) is one of the pathways of Rubisco exclusion from chloroplasts. To explore the relationship between the Rubisco contents and CPs under salt stress, Pn, maximum photochemical efficiency of PSII (F(v)/F(m)), carboxylation efficiency (CE) and concentration of Rubisco, number of CPs and Rubisco-containing Body (RCBs) were investigated with two differently salt-responding varieties in this experiment. We observed that 150 mM salt treatment resulted in not only significant decrease in Pn, CE and Rubisco content, but also obvious increase in the number of CPs and RCBs in salt-sensitive variety. Under salt stress formation of CPs resulted in production of much more RCBs, which could immigrate into and combine with vacuole. It may be a kind of important mechanism for rapid degradation of Rubisco under salt stress. Our conclusion provides a new sight for how Rubisco can be fast degraded under salt stress.

  7. Numerical prediction of the performance of horizontal sand filters as the pitch of the spiral protrusion changes.

    PubMed

    Mossad, Ruth; Aral, Hal

    2013-01-01

    The height of vertical sand filters are limited due to their structural cost. A horizontal sand filter may offer a better alternative; however, flow channels to the least resistance zone generated at the top of the filter as the sand gets wet and settles. A horizontal sand filter, internally baffled with spiral protrusion, is numerically modelled to study the effect of these spirals in reducing the channelling and enhancing the filter's effectiveness. Three different spiral pitches, 1.0, 0.75, and 0.5 m have been numerically modelled using Ansys FLUENT software. The parameters investigated were the power needed to run a flow rate through the horizontal filter and the residence time. The results show that as the spiral pitch decreases, the channelling reduces while the power increases. The power needed to pump a given flow rate of water in a 10 m long horizontal filter in all three cases investigated was less than the power needed to pump the same flow rate to the top of a 10 m long vertically standing sand filter. Results also showed that the time required for the flow to traverse through the sand filter increases in a nonlinear fashion as the pitch size decreases; however, the effectiveness of the filter increases. PMID:23676395

  8. High density infill in cracks and protrusions from the articular calcified cartilage in osteoarthritis in standardbred horse carpal bones.

    PubMed

    Laverty, Sheila; Lacourt, Mathieu; Gao, Chan; Henderson, Janet E; Boyde, Alan

    2015-01-01

    We studied changes in articular calcified cartilage (ACC) and subchondral bone (SCB) in the third carpal bones (C3) of Standardbred racehorses with naturally-occurring repetitive loading-induced osteoarthritis (OA). Two osteochondral cores were harvested from dorsal sites from each of 15 post-mortem C3 and classified as control or as showing early or advanced OA changes from visual inspection. We re-examined X-ray micro-computed tomography (µCT) image sets for the presence of high-density mineral infill (HDMI) in ACC cracks and possible high-density mineralized protrusions (HDMP) from the ACC mineralizing (tidemark) front (MF) into hyaline articular cartilage (HAC). We hypothesized and we show that 20-µm µCT resolution in 10-mm diameter samples is sufficient to detect HDMI and HDMP: these are lost upon tissue decalcification for routine paraffin wax histology owing to their predominant mineral content. The findings show that µCT is sufficient to discover HDMI and HDMP, which were seen in 2/10 controls, 6/9 early OA and 8/10 advanced OA cases. This is the first report of HDMI and HDMP in the equine carpus and in the Standardbred breed and the first to rely solely on µCT. HDMP are a candidate cause for mechanical tissue destruction in OA. PMID:25927581

  9. Interference of wedge-shaped protrusions on the faces of a Griffith crack in biaxial stress. Final report

    SciTech Connect

    Boulet, J.A.M.

    1992-04-01

    An initial investigation of the influence of protrusion interference on the fracture toughness required to prevent unstable propagation of a Griffith crack in a brittle material is described. The interference is caused by relative shear displacement of the crack faces when subjected to remote biaxial stress with neither principal stress parallel to the crack. It is shown that for room temperature cracks smaller than about one centimeter in silicon carbide, or about one millimeter in silicon nitride, the presence of interference changes the fracture stress. A mathematical model based on linear elasticity solutions and including multiple interference sites at arbitrarily specified positions on the crack is presented. Computations of the change in required fracture toughness and its dependence on wedge geometry (size and vertex angle), applied stresses (orientation and magnitude), and location of the interference site are discussed. Results indicate that a single interference site has only a slight effect on required toughness. However, the influence of interference increases monotonically with the number of interference sites. The two-dimensional model described herein is not accurate when the interference sites are closely spaced.

  10. Drosophila Dachsous and Fat polarize actin-based protrusions over a restricted domain of the embryonic denticle field.

    PubMed

    Lawlor, Kynan T; Ly, Daniel C; DiNardo, Stephen

    2013-11-15

    Atypical cadherins Dachsous (Ds) and Fat coordinate the establishment of planar polarity, essential for the patterning of complex tissues and organs. The precise mechanisms by which this system acts, particularly in cases where Ds and Fat act independently of the 'core' frizzled system, are still the subject of investigation. Examining the deployment of the Ds-Fat system in different tissues of the model organism Drosophila, has provided insights into the general mechanisms by which polarity is established and propagated to coordinate outcomes across a field of cells. The Drosophila embryonic epidermis provides a simple model epithelia where the establishment of polarity can be observed from start to finish, and in the absence of proliferation, over a fixed number of cells. Using the asymmetric placement of f-actin during denticle assembly as a read-out of polarity, we examine the requirement for Ds and Fat in establishing polarity across the denticle field. Comparing detailed phenotypic analysis with steady state protein enrichment revealed a spatially restricted requirement for the Ds-Fat system within the posterior denticle field. Ectopic Ds signaling provides evidence for a model whereby Ds acts to asymmetrically enrich Fat in a neighboring cell, in turn polarizing the cell to specify the position of the actin-based protrusions at the cell cortex.

  11. High Density Infill in Cracks and Protrusions from the Articular Calcified Cartilage in Osteoarthritis in Standardbred Horse Carpal Bones

    PubMed Central

    Laverty, Sheila; Lacourt, Mathieu; Gao, Chan; Henderson, Janet E.; Boyde, Alan

    2015-01-01

    We studied changes in articular calcified cartilage (ACC) and subchondral bone (SCB) in the third carpal bones (C3) of Standardbred racehorses with naturally-occurring repetitive loading-induced osteoarthritis (OA). Two osteochondral cores were harvested from dorsal sites from each of 15 post-mortem C3 and classified as control or as showing early or advanced OA changes from visual inspection. We re-examined X-ray micro-computed tomography (µCT) image sets for the presence of high-density mineral infill (HDMI) in ACC cracks and possible high-density mineralized protrusions (HDMP) from the ACC mineralizing (tidemark) front (MF) into hyaline articular cartilage (HAC). We hypothesized and we show that 20-µm µCT resolution in 10-mm diameter samples is sufficient to detect HDMI and HDMP: these are lost upon tissue decalcification for routine paraffin wax histology owing to their predominant mineral content. The findings show that µCT is sufficient to discover HDMI and HDMP, which were seen in 2/10 controls, 6/9 early OA and 8/10 advanced OA cases. This is the first report of HDMI and HDMP in the equine carpus and in the Standardbred breed and the first to rely solely on µCT. HDMP are a candidate cause for mechanical tissue destruction in OA. PMID:25927581

  12. High density infill in cracks and protrusions from the articular calcified cartilage in osteoarthritis in standardbred horse carpal bones.

    PubMed

    Laverty, Sheila; Lacourt, Mathieu; Gao, Chan; Henderson, Janet E; Boyde, Alan

    2015-04-28

    We studied changes in articular calcified cartilage (ACC) and subchondral bone (SCB) in the third carpal bones (C3) of Standardbred racehorses with naturally-occurring repetitive loading-induced osteoarthritis (OA). Two osteochondral cores were harvested from dorsal sites from each of 15 post-mortem C3 and classified as control or as showing early or advanced OA changes from visual inspection. We re-examined X-ray micro-computed tomography (µCT) image sets for the presence of high-density mineral infill (HDMI) in ACC cracks and possible high-density mineralized protrusions (HDMP) from the ACC mineralizing (tidemark) front (MF) into hyaline articular cartilage (HAC). We hypothesized and we show that 20-µm µCT resolution in 10-mm diameter samples is sufficient to detect HDMI and HDMP: these are lost upon tissue decalcification for routine paraffin wax histology owing to their predominant mineral content. The findings show that µCT is sufficient to discover HDMI and HDMP, which were seen in 2/10 controls, 6/9 early OA and 8/10 advanced OA cases. This is the first report of HDMI and HDMP in the equine carpus and in the Standardbred breed and the first to rely solely on µCT. HDMP are a candidate cause for mechanical tissue destruction in OA.

  13. Wnt/β-Catenin and Sonic Hedgehog Pathways Interact in the Regulation of the Development of the Dorsal Mesenchymal Protrusion

    PubMed Central

    Briggs, Laura E.; Burns, Tara A.; Lockhart, Marie M.; Phelps, Aimee L.; Van den Hoff, Maurice J.B.; Wessels, Andy

    2016-01-01

    Background The dorsal mesenchymal protrusion (DMP) is a second heart field (SHF) derived tissue involved in cardiac septation. Molecular mechanisms controlling SHF/DMP development include the Bone Morphogenetic Protein and Wnt/β-catenin signaling pathways. Reduced expression of components in these pathways leads to inhibition of proliferation of the SHF/DMP precursor population and failure of the DMP to develop. While the Sonic Hedgehog (Shh) pathway has also been demonstrated to be critically important for SHF/DMP development and atrioventricular septation, its role in the regulation of SHF proliferation is contentious. Results Tissue-specific deletion of the Shh receptor Smoothened from the SHF resulted in compromised DMP formation and atrioventricular septal defects (AVSDs). Immunohistochemical analysis at critical stages of DMP development showed significant proliferation defect as well as reduction in levels of the Wnt/β-catenin pathway-intermediates β-catenin, Lef1, and Axin2. To determine whether the defects seen in the conditional Smoothened knock-out mouse could be attributed to reduced Wnt/β-catenin signaling, LiCl, a pharmacological activator of this Wnt/β-catenin pathway, was administered. This resulted in restoration of proliferation and partial rescue of the AVSD phenotype. Conclusions The data presented suggest that the Wnt/β-catenin pathway interact with the Shh pathway in the regulation of SHF/DMP-precursor proliferation and, hence, the development of the DMP. PMID:26297872

  14. Reaching for far-flung antigen: How solid-core podosomes of dendritic cells transform into protrusive structures.

    PubMed

    Baranov, Maksim V; Ter Beest, Martin; van den Bogaart, Geert

    2014-10-01

    We recently identified a novel role for podosomes in antigen sampling. Podosomes are dynamic cellular structures that consist of point-like concentrations of actin surrounded by integrins and adaptor proteins such as vinculin and talin. Podosomes establish cellular contact with the extracellular matrix (ECM) and facilitate cell migration via ECM degradation. In our recent paper, we studied podosomes of human dendritic cells (DCs), major antigen presenting cells (APC) that take-up, process, and present foreign antigen to naive T-cells. We employed gelatin-impregnated porous polycarbonate filters to demonstrate that the mechanosensitive podosomes of DCs selectively localize to regions of low-physical resistance such as the filter pores. After degradation of the gelatin, podosomes increasingly protrude into the lumen of these pores. These protrusive podosome-derived structures contain several endocytic and early endosomal markers such as clathrin, Rab5, and VAMP3, and, surprisingly, also contain C-type lectins, a type of pathogen recognition receptors (PRRs). Finally, we performed functional uptake experiments to demonstrate that these PRRs facilitate uptake of antigen from the opposite side of the filter. Our data provide mechanistic insight in how dendritic cells sample for antigen across epithelial barriers for instance from the lumen of the lung and gut.

  15. Host cell polarity proteins participate in innate immunity to Pseudomonas aeruginosa infection

    PubMed Central

    Tran, Cindy S.; Eran, Yoni; Ruch, Travis R.; Bryant, David M.; Datta, Anirban; Brakeman, Paul; Kierbel, Arlinet; Wittmann, Torsten; Metzger, Ross J.; Mostov, Keith E.; Engel, Joanne N.

    2014-01-01

    Summary The mucosal epithelium consists of polarized cells with distinct apical and basolateral membranes that serve as functional and physical barriers to the organisms’ exterior. The apical surface of the epithelium constitutes the first point of contact between mucosal pathogens, such as Pseudomonas aeruginosa, and their host. We observed that binding of P. aeruginosa aggregates to the apical surface of polarized cells leads to the striking formation of an actin-rich membrane protrusion with ‘inverted’ polarity, containing basolateral lipids and membrane components. Such protrusions were associated with a spatially localized host immune response to P. aeruginosa aggregates that required bacterial flagella and a Type III secretion system apparatus. Host protrusions form de novo underneath bacterial aggregates and involve the apical recruitment of a Par3/Par6α/aPKC/Rac1 signaling module for a robust, spatially localized host NFκB response. Our data reveal an unanticipated role for spatio-temporal epithelial polarity changes in the activation of innate immune responses. PMID:24832456

  16. Method for Measuring the Distribution of Adhesion Forces on Continuous Nanoscale Protrusions Using Carbon Nanofiber Tip on a Scanning Probe Microscope Cantilever.

    PubMed

    Shimoi, Norihiro; Abe, Daisuke

    2015-07-01

    The adhesion force on surfaces has received attention in numerous scientific and technological fields, including catalysis, thin-film growth, and tribology. Many applications require knowledge of the strength of these forces as a function of position in three dimensions, but until now such information has only been theoretically proposed. Here, we demonstrate an approach based on scanning probe microscopy that can obtain such data and be used to image the three-dimensional surface force field of continuous nanoscale protrusions. We present adhesion force maps with nanometer and nanonewton resolution that allow detailed characterization of the interaction between a surface and a thin carbon nanofiber (CNF) rod synthesized by plasma-enhanced chemical vapor deposition (PECVD) at the end of a tip on a scanning probe microscope cantilever in three dimensions. In these maps, the positions of all continuous nanoscale protrusions are identified and the differences in the adhesive forces among limited areas at inequivalent sites are quantified.

  17. Membrane Processes.

    PubMed

    Pellegrin, Marie-Laure; Sadler, Mary E; Greiner, Anthony D; Aguinaldo, Jorge; Min, Kyungnan; Zhang, Kai; Arabi, Sara; Burbano, Marie S; Kent, Fraser; Shoaf, Robert

    2015-10-01

    This review, for literature published in 2014, contains information related to membrane processes for municipal and industrial applications. This review is a subsection of the Treatment Systems section of the annual Water Environment Federation literature review and covers the following topics: pretreatment, membrane bioreactor (MBR) configuration, design, nutrient removal, operation, industrial treatment, fixed film and anaerobic membrane systems, reuse, microconstituents removal, membrane technology advances, membrane fouling, and modeling. Other sub-sections of the Treatment Systems section that might relate to this literature review include: Biological Fixed-Film Systems, Activated Sludge and Other Aerobic Suspended Culture Processes, Anaerobic Processes, Water Reclamation and Reuse. The following sections might also have related information on membrane processes: Industrial Wastes, Hazardous Wastes, and Fate and Effects of Pollutants. PMID:26420079

  18. Membrane Processes.

    PubMed

    Pellegrin, Marie-Laure; Burbano, Marie S; Sadler, Mary E; Diamond, Jason; Baker, Simon; Greiner, Anthony D; Arabi, Sara; Wong, Joseph; Doody, Alexandra; Padhye, Lokesh P; Sears, Keith; Kistenmacher, Peter; Kent, Fraser; Tootchi, Leila; Aguinaldo, Jorge; Saddredini, Sara; Schilling, Bill; Min, Kyungnan; McCandless, Robert; Danker, Bryce; Gamage, Neranga P; Wang, Sunny; Aerts, Peter

    2016-10-01

    This review, for literature published in 2015, contains information related to membrane processes for municipal and industrial applications. This review is a subsection of the Treatment Systems section of the annual Water Environment Federation literature review and covers the following topics: pretreatment, membrane bioreactor (MBR) configuration, design, nutrient removal, operation, industrial treatment, anaerobic membrane systems, reuse, microconstituents removal, membrane technology advances, membrane fouling, and modeling. Other sub-sections of the Treatment Systems section that might relate to this literature review include: Biological Fixed-Film Systems, Activated Sludge and Other Aerobic Suspended Culture Processes, Anaerobic Processes, Water Reclamation and Reuse. The following sections might also have related information on membrane processes: Industrial Wastes, Hazardous Wastes, and Fate and Effects of Pollutants. PMID:27620084

  19. Multicomponent membranes

    DOEpatents

    Kulprathipanja, Santi; Kulkarni, Sudhir S.; Funk, Edward W.

    1988-01-01

    A multicomponent membrane which may be used for separating various components which are present in a fluid feed mixture comprises a mixture of a plasticizer such as a glycol and an organic polymer cast upon a porous organic polymer support. The membrane may be prepared by casting an emulsion or a solution of the plasticizer and polymer on the porous support, evaporating the solvent and recovering the membrane after curing.

  20. T-Stenting-and-Small-Protrusion Technique for Bifurcation Stenoses After End-to-Side Anastomosis of Transplant Renal Artery and External Iliac Artery: Report of Two Cases

    SciTech Connect

    Chen, Yong Ye, Peng; Jiang, Wen-jin; Ma, Shuo-yi Zhao, Jian-bo Zeng, Qing-le

    2015-10-15

    Bifurcation stenoses after end-to-side anastomosis of transplant renal artery (TRA) and external iliac artery (EIA), including stenoses at the anastomosis and the iliac artery proximal to the TRA, are rare. In the present article, we report two successfully managed cases of bifurcation stenoses after end-to-side anastomosis of the TRA and EIA using the technique of T-stenting and small protrusion (TAP stenting)

  1. Charged membranes.

    PubMed

    Thatcher, Jack D

    2013-04-16

    This Teaching Resource provides three animated lessons that describe the storage and utilization of energy across plasma membranes. The "Na,K ATPase" animation explains how these pumps establish the electrochemical gradient that stores energy across plasma membranes. The "ATP synthesizing complexes" animation shows how these complexes transfer energy from the inner mitochondrial membrane to adenosine triphosphate (ATP). The "action potential" lesson explains how charged membranes are used to propagate signals along the axons of neurons. These animations serve as valuable resources for any collegiate-level course that describes these important factors. Courses that might employ them include introductory biology, biochemistry, biophysics, cell biology, pharmacology, and physiology.

  2. MYADM regulates Rac1 targeting to ordered membranes required for cell spreading and migration.

    PubMed

    Aranda, Juan F; Reglero-Real, Natalia; Kremer, Leonor; Marcos-Ramiro, Beatriz; Ruiz-Sáenz, Ana; Calvo, María; Enrich, Carlos; Correas, Isabel; Millán, Jaime; Alonso, Miguel A

    2011-04-15

    Membrane organization into condensed domains or rafts provides molecular platforms for selective recruitment of proteins. Cell migration is a general process that requires spatiotemporal targeting of Rac1 to membrane rafts. The protein machinery responsible for making rafts competent to recruit Rac1 remains elusive. Some members of the MAL family of proteins are involved in specialized processes dependent on this type of membrane. Because condensed membrane domains are a general feature of the plasma membrane of all mammalian cells, we hypothesized that MAL family members with ubiquitous expression and plasma membrane distribution could be involved in the organization of membranes for cell migration. We show that myeloid-associated differentiation marker (MYADM), a protein with unique features within the MAL family, colocalizes with Rac1 in membrane protrusions at the cell surface and distributes in condensed membranes. MYADM knockdown (KD) cells had altered membrane condensation and showed deficient incorporation of Rac1 to membrane raft fractions and, similar to Rac1 KD cells, exhibited reduced cell spreading and migration. Results of rescue-of-function experiments by expression of MYADM or active Rac1L61 in cells knocked down for Rac1 or MYADM, respectively, are consistent with the idea that MYADM and Rac1 act on parallel pathways that lead to similar functional outcomes. PMID:21325632

  3. Formation of membrane ridges and scallops by the F-BAR protein Nervous Wreck

    PubMed Central

    Becalska, Agata N.; Kelley, Charlotte F.; Berciu, Cristina; Stanishneva-Konovalova, Tatiana B.; Fu, Xiaofeng; Wang, ShiYu; Sokolova, Olga S.; Nicastro, Daniela; Rodal, Avital A.

    2013-01-01

    Eukaryotic cells are defined by extensive intracellular compartmentalization, which requires dynamic membrane remodeling. FER/Cip4 homology-Bin/amphiphysin/Rvs (F-BAR) domain family proteins form crescent-shaped dimers, which can bend membranes into buds and tubules of defined geometry and lipid composition. However, these proteins exhibit an unexplained wide diversity of membrane-deforming activities in vitro and functions in vivo. We find that the F-BAR domain of the neuronal protein Nervous Wreck (Nwk) has a novel higher-order structure and membrane-deforming activity that distinguishes it from previously described F-BAR proteins. The Nwk F-BAR domain assembles into zigzags, creating ridges and periodic scallops on membranes in vitro. This activity depends on structural determinants at the tips of the F-BAR dimer and on electrostatic interactions of the membrane with the F-BAR concave surface. In cells, Nwk-induced scallops can be extended by cytoskeletal forces to produce protrusions at the plasma membrane. Our results define a new F-BAR membrane-deforming activity and illustrate a molecular mechanism by which positively curved F-BAR domains can produce a variety of membrane curvatures. These findings expand the repertoire of F-BAR domain mediated membrane deformation and suggest that unique modes of higher-order assembly can define how these proteins sculpt the membrane. PMID:23761074

  4. Randomized clinical trial comparing lumbar percutaneous hydrodiscectomy with lumbar open microdiscectomy for the treatment of lumbar disc protrusions and herniations

    PubMed Central

    Cristante, Alexandre Fogaça; Rocha, Ivan Diasda; Marcon, Raphael Martus; de Barros Filho, Tarcísio Eloy Pessoa

    2016-01-01

    OBJECTIVES: Hydrodiscectomy is a new technique used for percutaneous spinal discectomy that employs a high-intensity stream of water for herniated disc ablation and tissue aspiration. No previous clinical study has examined the effects of percutaneous hydrodiscectomy. The aim of this study is to evaluate the outcomes of hydrodiscectomy compared to open microdiscectomy regarding pain, function, satisfaction, complications and recurrence rates. METHODS: In this randomized clinical trial, patients referred to our tertiary hospital for lumbar back pain were recruited and included in the study if they had disc protrusion or small herniation in only one level, without neurological deficits and with no resolution after six weeks of conservative treatment. One group underwent open microdiscectomy, and the other group underwent percutaneous microdiscectomy via hydrosurgery. Function was evaluated using the Oswestry Disability Index and pain was assessed using a visual analog scale. Evaluations were performed preoperatively, and then during the first week and at one, three, six and twelve months postoperatively. Personal satisfaction was verified. Clinicaltrials.gov: NCT01367860. RESULTS: During the study period, 20 patients were included in each arm and 39 completed one-year of follow-up (one patient died of unrelated causes). Both groups exhibited equal improvement on the visual analog scale and Oswestry evaluations after treatment, without any significant differences. The improvement in the lumbar visual analog scale score was not significant in the hydrodiscectomy group (p=0.138). The rates of infection, pain, recurrence and satisfaction were similar between the two groups. CONCLUSION: Percutaneous hydrodiscectomy was demonstrated to be as effective as open microdiscectomy for reducing pain. The rates of complications and recurrence of herniation were similar between groups. Patient satisfaction with the treatment was also similar between groups. PMID:27276397

  5. Feedback regulation between plasma membrane tension and membrane-bending proteins organizes cell polarity during leading edge formation.

    PubMed

    Tsujita, Kazuya; Takenawa, Tadaomi; Itoh, Toshiki

    2015-06-01

    Tension applied to the plasma membrane (PM) is a global mechanical parameter involved in cell migration. However, how membrane tension regulates actin assembly is unknown. Here, we demonstrate that FBP17, a membrane-bending protein and an activator of WASP/N-WASP-dependent actin nucleation, is a PM tension sensor involved in leading edge formation. In migrating cells, FBP17 localizes to short membrane invaginations at the leading edge, while diminishing from the cell rear in response to PM tension increase. Conversely, following reduced PM tension, FBP17 dots randomly distribute throughout the cell, correlating with loss of polarized actin assembly on PM tension reduction. Actin protrusive force is required for the polarized accumulation, indicating a role for FBP17-mediated activation of WASP/N-WASP in PM tension generation. In vitro experiments show that FBP17 membrane-bending activity depends on liposomal membrane tension. Thus, FBP17 is the local activator of actin polymerization that is inhibited by PM tension in the feedback loop that regulates cell migration.

  6. Crystalline Membranes

    NASA Technical Reports Server (NTRS)

    Tsapatsis, Michael (Inventor); Lai, Zhiping (Inventor)

    2008-01-01

    In certain aspects, the invention features methods for forming crystalline membranes (e.g., a membrane of a framework material, such as a zeolite) by inducing secondary growth in a layer of oriented seed crystals. The rate of growth of the seed crystals in the plane of the substrate is controlled to be comparable to the rate of growth out of the plane. As a result, a crystalline membrane can form a substantially continuous layer including grains of uniform crystallographic orientation that extend through the depth of the layer.

  7. Dynamin and endocytosis are required for the fusion of osteoclasts and myoblasts

    PubMed Central

    Shin, Nah-Young; Choi, Hyewon; Neff, Lynn; Wu, Yumei; Saito, Hiroaki; Ferguson, Shawn M.; De Camilli, Pietro

    2014-01-01

    Cell–cell fusion is an evolutionarily conserved process that leads to the formation of multinucleated myofibers, syncytiotrophoblasts and osteoclasts, allowing their respective functions. Although cell–cell fusion requires the presence of fusogenic membrane proteins and actin-dependent cytoskeletal reorganization, the precise machinery allowing cells to fuse is still poorly understood. Using an inducible knockout mouse model to generate dynamin 1– and 2–deficient primary osteoclast precursors and myoblasts, we found that fusion of both cell types requires dynamin. Osteoclast and myoblast cell–cell fusion involves the formation of actin-rich protrusions closely associated with clathrin-mediated endocytosis in the apposed cell. Furthermore, impairing endocytosis independently of dynamin also prevented cell–cell fusion. Since dynamin is involved in both the formation of actin-rich structures and in endocytosis, our results indicate that dynamin function is central to the osteoclast precursors and myoblasts fusion process, and point to an important role of endocytosis in cell–cell fusion. PMID:25287300

  8. Tks5-dependent, Nox-mediated Generation of Reactive Oxygen Species is Necessary for Invadopodia Formation*

    PubMed Central

    Diaz, Begoña; Shani, Gidon; Pass, Ian; Anderson, Diana; Quintavalle, Manuela; Courtneidge, Sara A.

    2009-01-01

    Invadopodia are actin-rich membrane protrusions of cancer cells which facilitate pericellular proteolysis and invasive behavior. We show here that reactive oxygen species (ROS) generated by the NADPH oxidase (Nox) system are necessary for invadopodia formation and function. The invadopodia protein Tks5 is structurally related to p47phox, a Nox component in phagocytic cells. Knockdown of Tks5 reduces total ROS levels in cancer cells. Furthermore, Tks5 and p22phox can associate with each other, suggesting that Tks5 is part of the Nox complex. Tyrosine phosphorylation of Tks5 and Tks4, but not other Src substrates, is reduced by Nox inhibition. We propose that Tks5 facilitates the production of ROS necessary for invadopodia formation, and that in turn ROS modulates Tks5 tyrosine phosphorylation in a positive feedback loop. PMID:19755709

  9. Tracing myoblast fusion in Drosophila embryos by fluorescent actin probes.

    PubMed

    Haralalka, Shruti; Abmayr, Susan M

    2015-01-01

    Myoblast fusion in the Drosophila embryo is a highly elaborate process that is initiated by Founder Cells and Fusion-Competent Myoblasts (FCMs). It occurs through an asymmetric event in which actin foci assemble in the FCMs at points of cell-cell contact and direct the formation of membrane protrusions that drive fusion. Herein, we describe the approach that we have used to image in living embryos the highly dynamic actin foci and actin-rich projections that precede myoblast fusion. We discuss resources currently available for imaging actin and myogenesis, and our experience with these resources if available. This technical report is not intended to be comprehensive on providing instruction on standard microscopy practices or software utilization. However, we discuss microscope parameters that we have used in data collection, and our experience with image processing tools in data analysis.

  10. Biological membranes

    PubMed Central

    Watson, Helen

    2015-01-01

    Biological membranes allow life as we know it to exist. They form cells and enable separation between the inside and outside of an organism, controlling by means of their selective permeability which substances enter and leave. By allowing gradients of ions to be created across them, membranes also enable living organisms to generate energy. In addition, they control the flow of messages between cells by sending, receiving and processing information in the form of chemical and electrical signals. This essay summarizes the structure and function of membranes and the proteins within them, and describes their role in trafficking and transport, and their involvement in health and disease. Techniques for studying membranes are also discussed. PMID:26504250

  11. Membrane Nanotubes

    NASA Astrophysics Data System (ADS)

    Derényi, I.; Koster, G.; van Duijn, M. M.; Czövek, A.; Dogterom, M.; Prost, J.

    There is a growing pool of evidence showing the biological importance of membrane nanotubes (with diameter of a few tens of nanometers and length upto tens of microns) in various intra- and intercellular transport processes. These ubiquitous structures are often formed from flat membranes by highly localized forces generated by either the pulling of motor proteins or the pushing of polymerizing cytoskeletal filaments. In this chapter we give an overview of the theory of membrane nanotubes, their biological relevance, and the most recent experiments designed for the study of their formation and dynamics. We also discuss the effect of membrane proteins or lipid composition on the shape of the tubes, and the effect of antagonistic motor proteins on tube formation.

  12. Macrophages engulf endothelial cell membrane particles preceding pupillary membrane capillary regression

    PubMed Central

    Poché, Ross A.; Hsu, Chih-Wei; McElwee, Melissa L.; Burns, Alan R.; Dickinson, Mary E.

    2015-01-01

    Programmed capillary regression and remodeling are essential developmental processes. However, the cellular and molecular mechanisms that regulate vessel regression are only beginning to be understood. Here, using in vivo, dynamic, confocal imaging of mouse transgenic reporters as well as static confocal and electron microscopy, we studied the embryonic development and postnatal regression of the transient mouse pupillary membrane (PM) vasculature. This approach allowed us to directly observe the precise temporal sequence of cellular events preceding and during the elimination of the PM from the mouse eye. Imaging of Tcf/Lef-H2B::GFP Wnt-reporter mice uncovered that, unlike the hyaloid vasculature of the posterior eye, a PM endothelial cell (EC) Wnt/β-catenin response is unlikely to be part of the regression mechanism. Live imaging of EC and macrophage dynamics revealed highly active Csf1r-GFP+ macrophages making direct contact with the Flk1-myr::mCherry+ vessel surface and with membrane protrusions or filopodia extending from the ECs. Flk1-myr::mCherry+ EC membrane particles were observed on and around ECs as well as within macrophages. Electron microscopy studies confirmed that they were in phagosomes within macrophages, indicating that the macrophages engulfed the membrane particles. Interestingly, EC plasma membrane uptake by PM macrophages did not correlate with apoptosis and was found shortly after vessel formation at mid-gestation stages in the embryo; long before vessel regression begins during postnatal development. Additionally, genetic ablation of macrophages showed that EC membrane particles were still shed in the absence of macrophages suggesting that macrophages do not induce the formation or release of EC microparticles. These studies have uncovered a novel event during programmed capillary regression in which resident macrophages scavenge endothelial cell microparticles released from the PM vessels. This finding suggests that there may be an

  13. Protrusive intrusion, dehydration and polymorphism in minerals as possible reason of seismic activity, relation between ophiolite belts and seismic zonation of the territory of Armenia

    NASA Astrophysics Data System (ADS)

    Harutyunyan, A. V.; Petrosyan, H. M.

    2010-05-01

    In the basis of multiple geological and geophysical data, also on the results of investigations seismic and density properties of rocks at high termobaric conditions, we proposed the petrophisical section and model of evolution of Earth crust of the territory of Armenia. On the proposed model the following interrelated problems are debated: forming of ophiolite belts and volcanic centers, genesis of hydrocarbons by organic and inorganic ways, and also reasons of originating of seismic centers. The reasons of originating of seismic centers in different depths of Earth crust, are miscellaneous. According to the model of Earth crust evolution the ophiolite belts are formed due to permanent protrusive intrusion of serpentinized masses from the foot of the crust (35-50km) into upper horizons. It is natural to assume, that the permanent intrusion of serpentinizd masses through deep faults has drastically occurred accompanying with seismic shakings. This process encourages the development of deep faults. The protrusive intrusion of serpentinized masse accompanied with partial dehydration of serpentinites and serpentinized ultrabasites and new mineral formation. The processes was accompanied also with drastic change of seismic waves and volumes up to 30%. Experiments at high termobaric conditions show, that some minerals undergone polymorphous transformations, accompanied with phase change and drastic change of rocks volume. Particularly plastic calcite, included in the composition of metamorphic rocks to run into the cracks expends and diversifies them. The process described cause some general effects similar to those of the process of dilatancy. Therefore, the protrusive intrusion of serpentinized masses into upper horizons, it dehydrations and polymorphous transformations in different minerals, may be cause of geo-dynamic processes at different depths of Earth crust. It may be assumed, that those processes permanently occur nowadays as well. Comparing the maps of

  14. Mechanisms of Membrane Binding of Small GTPase K-Ras4B Farnesylated Hypervariable Region*

    PubMed Central

    Jang, Hyunbum; Abraham, Sherwin J.; Chavan, Tanmay S.; Hitchinson, Ben; Khavrutskii, Lyuba; Tarasova, Nadya I.; Nussinov, Ruth; Gaponenko, Vadim

    2015-01-01

    K-Ras4B belongs to a family of small GTPases that regulates cell growth, differentiation and survival. K-ras is frequently mutated in cancer. K-Ras4B association with the plasma membrane through its farnesylated and positively charged C-terminal hypervariable region (HVR) is critical to its oncogenic function. However, the structural mechanisms of membrane association are not fully understood. Here, using confocal microscopy, surface plasmon resonance, and molecular dynamics simulations, we observed that K-Ras4B can be distributed in rigid and loosely packed membrane domains. Its membrane binding domain interaction with phospholipids is driven by membrane fluidity. The farnesyl group spontaneously inserts into the disordered lipid microdomains, whereas the rigid microdomains restrict the farnesyl group penetration. We speculate that the resulting farnesyl protrusion toward the cell interior allows oligomerization of the K-Ras4B membrane binding domain in rigid microdomains. Unlike other Ras isoforms, K-Ras4B HVR contains a single farnesyl modification and positively charged polylysine sequence. The high positive charge not only modulates specific HVR binding to anionic phospholipids but farnesyl membrane orientation. Phosphorylation of Ser-181 prohibits spontaneous farnesyl membrane insertion. The mechanism illuminates the roles of HVR modifications in K-Ras4B targeting microdomains of the plasma membrane and suggests an additional function for HVR in regulation of Ras signaling. PMID:25713064

  15. Vocal anatomy, tongue protrusion behaviour and the acoustics of rutting roars in free-ranging Iberian red deer stags (Cervus elaphus hispanicus)

    PubMed Central

    Frey, Roland; Volodin, Ilya; Volodina, Elena; Carranza, Juan; Torres-Porras, Jerónimo

    2012-01-01

    Roaring in rutting Iberian red deer stags Cervus elaphus hispanicus is unusual compared to other subspecies of red deer, which radiated from the Iberian refugium after the last glacial maximum. In all red deer stags, the larynx occupies a permanent low mid-neck resting position and is momentarily retracted almost down to the rostral end of the sternum during the production of rutting calls. Simultaneous with the retraction of the larynx, male Iberian red deer pronouncedly protrude the tongue during most of their rutting roars. This poses a mechanical challenge for the vocal tract (vt) and for the hyoid apparatus, as tongue and larynx are strongly pulled in opposite directions. This study (i) examines the vocal anatomy and the acoustics of the rutting roars in free-ranging male C. e. hispanicus; (ii) establishes a potential mechanism of simultaneous tongue protrusion and larynx retraction by applying a two-dimensional model based on graphic reconstructions in single video frames of unrestrained animals; and (iii) advances a hypothesis of evaporative cooling by tongue protrusion in the males of a subspecies of red deer constrained to perform all of the exhausting rutting activities, including acoustic display, in a hot and arid season. PMID:22257361

  16. Formation of chloroplast protrusions and catalase activity in alpine Ranunculus glacialis under elevated temperature and different CO2/O2 ratios.

    PubMed

    Buchner, Othmar; Moser, Tim; Karadar, Matthias; Roach, Thomas; Kranner, Ilse; Holzinger, Andreas

    2015-11-01

    Chloroplast protrusions (CPs) have frequently been observed in plants, but their significance to plant metabolism remains largely unknown. We investigated in the alpine plant Ranunculus glacialis L. treated under various CO2 concentrations if CP formation is related to photorespiration, specifically focusing on hydrogen peroxide (H2O2) metabolism. Immediately after exposure to different CO2 concentrations, the formation of CPs in leaf mesophyll cells was assessed and correlated to catalase (CAT) and ascorbate peroxidase (APX) activities. Under natural irradiation, the relative proportion of chloroplasts with protrusions (rCP) was highest (58.7 %) after exposure to low CO2 (38 ppm) and was lowest (3.0 %) at high CO2 (10,000 ppm). The same relationship was found for CAT activity, which decreased from 34.7 nkat mg(-1) DW under low CO2 to 18.4 nkat mg(-1) DW under high CO2, while APX activity did not change significantly. When exposed to natural CO2 concentration (380 ppm) in darkness, CP formation was significantly lower (18.2 %) compared to natural solar irradiation (41.3 %). In summary, CP formation and CAT activity are significantly increased under conditions that favour photorespiration, while in darkness or at high CO2 concentration under light, CP formation is significantly lower, providing evidence for an association between CPs and photorespiration.

  17. Broadening the Spectrum of Actin-Based Protrusive Activity Mediated by Arp2/3 Complex-Facilitated Polymerization: Motility of Cytoplasmic Ridges and Tubular Projections

    PubMed Central

    Henson, John H.; Gianakas, Anastasia D.; Henson, Lauren H.; Lakin, Christina L.; Voss, Meagen K.; Bewersdorf, Joerg; Oldenbourg, Rudolf; Morris, Robert L.

    2014-01-01

    Arp2/3 complex-facilitated actin polymerization plays an essential role in a variety of cellular functions including motility, adherence, endocytosis and trafficking. In the present study we employ the sea urchin coelomocyte experimental model system to test the hypotheses that Arp2/3 complex-nucleated actin assembly mediates the motility of two unusual cellular protrusions; the cytoplasmic ridges present during coelomocyte spreading, and inducible, tubular-shaped, and neurite-like projections. Our investigations couple pharmacological manipulation employing inhibitors of actin polymerization and the Arp2/3 complex with a wide array of imaging methods including digitally enhanced phase contrast, DIC and polarization light microscopy of live cells; conventional, confocal and super-resolution light microscopy of fluorescently labeled cells; and scanning and transmission electron microscopy. Taken together, the results of this study indicate that Arp2/3 complex-facilitated actin polymerization underlies the motility of coelomocyte cytoplasmic ridges and tubular projections, that these processes are related to each other, and that they have been preliminarily identified in other cell types. The results also highlight the broad spectrum of actin-based protrusive activities dependent on the Arp2/3 complex and provide additional insights into the pervasive nature of this ubiquitous actin nucleator. Furthermore we provide the first evidence of a possible mechanistic difference between the impacts of the small molecule drugs BDM and CK666 on the Arp2/3 complex. PMID:25111797

  18. Membrane magic

    SciTech Connect

    Buecker, B.

    2005-09-01

    The Kansas Power and Light Co.'s La Cyne generating station has found success with membrane filtration water pretreatment technology. The article recounts the process followed in late 2004 to install a Pall Aria 4 microfilter in Unit 1 makeup water system at the plant to produce cleaner water for reverse osmosis feed. 2 figs., 2 photos.

  19. Self-assembly of 2D membranes from mixtures of hard rods and depleting polymers().

    PubMed

    Yang, Yasheng; Barry, Edward; Dogic, Zvonimir; Hagan, Michael F

    2012-01-01

    We combine simulations and experiments to elucidate the molecular forces leading to the assembly of two dimensional membrane-like structures composed of a one rod-length thick monolayer of aligned rods from an immiscible suspension of hard rods and depleting polymers. Computer simulations predict that monolayer membranes are thermodynamically stable above a critical rod aspect ratio and below a critical depletion interaction length scale. Outside of these conditions alternative structures such as stacked smectic columns or nematic droplets are thermodynamically stable. These predictions are confirmed by subsequent experiments using a model system of virus rod-like molecules and non-adsorbing polymer. Our work demonstrates that collective molecular protrusion fluctuations alone are sufficient to stabilize membranes composed of homogenous rods with simple excluded volume interactions.

  20. F-actin-rich contractile endothelial pores prevent vascular leakage during leukocyte diapedesis through local RhoA signalling.

    PubMed

    Heemskerk, Niels; Schimmel, Lilian; Oort, Chantal; van Rijssel, Jos; Yin, Taofei; Ma, Bin; van Unen, Jakobus; Pitter, Bettina; Huveneers, Stephan; Goedhart, Joachim; Wu, Yi; Montanez, Eloi; Woodfin, Abigail; van Buul, Jaap D

    2016-01-27

    During immune surveillance and inflammation, leukocytes exit the vasculature through transient openings in the endothelium without causing plasma leakage. However, the exact mechanisms behind this intriguing phenomenon are still unknown. Here we report that maintenance of endothelial barrier integrity during leukocyte diapedesis requires local endothelial RhoA cycling. Endothelial RhoA depletion in vitro or Rho inhibition in vivo provokes neutrophil-induced vascular leakage that manifests during the physical movement of neutrophils through the endothelial layer. Local RhoA activation initiates the formation of contractile F-actin structures that surround emigrating neutrophils. These structures that surround neutrophil-induced endothelial pores prevent plasma leakage through actomyosin-based pore confinement. Mechanistically, we found that the initiation of RhoA activity involves ICAM-1 and the Rho GEFs Ect2 and LARG. In addition, regulation of actomyosin-based endothelial pore confinement involves ROCK2b, but not ROCK1. Thus, endothelial cells assemble RhoA-controlled contractile F-actin structures around endothelial pores that prevent vascular leakage during leukocyte extravasation.

  1. F-actin-rich contractile endothelial pores prevent vascular leakage during leukocyte diapedesis through local RhoA signalling

    PubMed Central

    Heemskerk, Niels; Schimmel, Lilian; Oort, Chantal; van Rijssel, Jos; Yin, Taofei; Ma, Bin; van Unen, Jakobus; Pitter, Bettina; Huveneers, Stephan; Goedhart, Joachim; Wu, Yi; Montanez, Eloi; Woodfin, Abigail; van Buul, Jaap D.

    2016-01-01

    During immune surveillance and inflammation, leukocytes exit the vasculature through transient openings in the endothelium without causing plasma leakage. However, the exact mechanisms behind this intriguing phenomenon are still unknown. Here we report that maintenance of endothelial barrier integrity during leukocyte diapedesis requires local endothelial RhoA cycling. Endothelial RhoA depletion in vitro or Rho inhibition in vivo provokes neutrophil-induced vascular leakage that manifests during the physical movement of neutrophils through the endothelial layer. Local RhoA activation initiates the formation of contractile F-actin structures that surround emigrating neutrophils. These structures that surround neutrophil-induced endothelial pores prevent plasma leakage through actomyosin-based pore confinement. Mechanistically, we found that the initiation of RhoA activity involves ICAM-1 and the Rho GEFs Ect2 and LARG. In addition, regulation of actomyosin-based endothelial pore confinement involves ROCK2b, but not ROCK1. Thus, endothelial cells assemble RhoA-controlled contractile F-actin structures around endothelial pores that prevent vascular leakage during leukocyte extravasation. PMID:26814335

  2. Distinct Roles for the N- and C-terminal Regions of M-Sec in Plasma Membrane Deformation during Tunneling Nanotube Formation

    PubMed Central

    Kimura, Shunsuke; Yamashita, Masami; Yamakami-Kimura, Megumi; Sato, Yusuke; Yamagata, Atsushi; Kobashigawa, Yoshihiro; Inagaki, Fuyuhiko; Amada, Takako; Hase, Koji; Iwanaga, Toshihiko; Ohno, Hiroshi; Fukai, Shuya

    2016-01-01

    The tunneling nanotube (TNT) is a structure used for intercellular communication, and is a thin membrane protrusion mediating transport of various signaling molecules and cellular components. M-Sec has potent membrane deformation ability and induces TNT formation in cooperation with the Ral/exocyst complex. Here, we show that the N-terminal polybasic region of M-Sec directly binds phosphatidylinositol (4,5)-bisphosphate for its localization to the plasma membrane during the initial stage of TNT formation. We further report a crystal structure of M-Sec, which consists of helix bundles arranged in a straight rod-like shape, similar to the membrane tethering complex subunits. A positively charged surface in the C-terminal domains is required for M-Sec interaction with active RalA to extend the plasma membrane protrusions. Our results suggest that the membrane-associated M-Sec recruits active RalA, which directs the exocyst complex to form TNTs. PMID:27629377

  3. Distinct Roles for the N- and C-terminal Regions of M-Sec in Plasma Membrane Deformation during Tunneling Nanotube Formation.

    PubMed

    Kimura, Shunsuke; Yamashita, Masami; Yamakami-Kimura, Megumi; Sato, Yusuke; Yamagata, Atsushi; Kobashigawa, Yoshihiro; Inagaki, Fuyuhiko; Amada, Takako; Hase, Koji; Iwanaga, Toshihiko; Ohno, Hiroshi; Fukai, Shuya

    2016-01-01

    The tunneling nanotube (TNT) is a structure used for intercellular communication, and is a thin membrane protrusion mediating transport of various signaling molecules and cellular components. M-Sec has potent membrane deformation ability and induces TNT formation in cooperation with the Ral/exocyst complex. Here, we show that the N-terminal polybasic region of M-Sec directly binds phosphatidylinositol (4,5)-bisphosphate for its localization to the plasma membrane during the initial stage of TNT formation. We further report a crystal structure of M-Sec, which consists of helix bundles arranged in a straight rod-like shape, similar to the membrane tethering complex subunits. A positively charged surface in the C-terminal domains is required for M-Sec interaction with active RalA to extend the plasma membrane protrusions. Our results suggest that the membrane-associated M-Sec recruits active RalA, which directs the exocyst complex to form TNTs. PMID:27629377

  4. Membrane tension homeostasis of epithelial cells through surface area regulation in response to osmotic stress.

    PubMed

    Pietuch, Anna; Brückner, Bastian R; Janshoff, Andreas

    2013-03-01

    Osmotic stress poses one of the most fundamental challenges to living cells. Particularly, the largely inextensible plasma membrane of eukaryotic cells easily ruptures under in-plane tension calling for sophisticated strategies to readily respond to osmotic stress. We describe how epithelial cells react and adapt mechanically to the exposure to hypotonic and hypertonic solutions in the context of a confluent monolayer. Site-specific indentation experiments in conjunction with tether pulling on individual cells have been carried out with an atomic force microscope to reveal spatio-temporal changes in membrane tension and surface area. We found that cells compensate for an increase in lateral tension due to hypoosmotic stress by sacrificing excess of membrane area stored in protrusions and invaginations such as microvilli and caveolae. At mild hypotonic conditions lateral tension increases partly compensated by surface are regulation, i.e. the cell sacrifices some of its membrane reservoirs. A loss of membrane-actin contacts occurs upon exposure to stronger hypotonic solutions giving rise to a drop in lateral tension. Tension release recovers on longer time scales by an increasing endocytosis, which efficiently removes excess membrane from the apical side to restore the initial pre-stress. Hypertonic solutions lead to shrinkage of cells and collapse of the apical membrane onto the cortex. Exposure to distilled water leads to stiffening of cells due to removal of excess surface area and tension increase due to elevated osmotic pressure across the plasma membrane.

  5. [Membranous nephropathy].

    PubMed

    Mercadal, Lucile

    2013-12-01

    Membranous nephropathy is characterized by immune complex deposits on the outer side of the glomerular basement membrane. Activation of complement and of oxidation lead to basement membrane lesions. The most frequent form is idiopathic. At 5 and 10 years, renal survival is around 90 and 65% respectively. A prognostic model based on proteinuria, level and duration, progression of renal failure in a few months can refine prognosis. The urinary excretion of C5b-9, β2 and α1 microglobuline and IgG are strong predictors of outcome. Symptomatic treatment is based on anticoagulation in case of nephrotic syndrome, angiotensin conversion enzyme inhibitors, angiotensin II receptor blockers and statins. Immunosuppressive therapy should be discussed for patients having a high risk of progression. Corticoids alone has no indication. Treatment should include a simultaneous association or more often alternating corticoids and alkylant agent for a minimum of 6 months. Adrenocorticoid stimulating hormone and steroids plus mycophenolate mofetil may be equally effective. Steroids plus alkylant decrease the risk of end stage renal failure. Cyclosporine and tacrolimus decrease proteinuria but are associated with a high risk of recurrence at time of withdrawal and are nephrotoxic. Rituximab evaluated on open studies needs further evaluations to define its use.

  6. Characterization of moesin in the sea urchin Lytechinus variegatus: redistribution to the plasma membrane following fertilization is inhibited by cytochalasin B.

    PubMed

    Bachman, E S; McClay, D R

    1995-01-01

    We have investigated the distribution and function of an ezrin-radixin-moesin-like (ERM) molecule in the sea urchin. A sea urchin homologue of moesin was cloned that shares 75% amino acid similarity in the conserved N-terminal region to other moesin molecules. A 6.3 kb message is transcribed late in embryogenesis and is present in adult tissues. Polyclonal antibodies were generated to proteins expressed by a bacterial expression vector, and affinity purified. These antibodies recognize a single 75 kDa protein that is present throughout development in approximately equal abundance, and specifically they immuno-precipitate a single protein. We show by immunolocalization that SUmoesin has two predominant patterns during development. First, SUmoesin is rapidly redistributed after fertilization from a location throughout the egg cytoplasm to a location in the egg cortex. Later in embryogenesis, SUmoesin is localized to the apical ends of cells in the regions of cell-cell junctions. We show that SUmoesin is present in actin-rich regions of the embryo. Finally, we show that the location of SUmoesin requires an intact actin-based cytoskeleton. SUmoesin fails to localize to the plasma membrane after fertilization in the presence of cytochalasin B. Furthermore, SUmoesin loses its apical position in the region of cell-cell junctions in the presence of cytochalasin B in later stages of embryogenesis. This effect is reversible, and the microtubule inhibitor colchicine has no effect. These results show that SUmoesin becomes associated with apical plasma membrane structures early in development, and that SUmoesin is both coincident with actin and requires the assembly of actin filaments to maintain its localization. PMID:7738093

  7. Effects of reversible inactivation by cooling of the primate face motor cortex on the performance of a trained tongue-protrusion task and a trained biting task.

    PubMed

    Murray, G M; Lin, L D; Moustafa, E M; Sessle, B J

    1991-03-01

    1. Intracortical microstimulation (ICMS) and surface stimulation studies of primate face motor cortex have shown an extensive representation within face motor cortex devoted to movements of the tongue and face; only a very small representation for jaw-closing movements has ever been demonstrated. These data suggest that face motor cortex plays a critical role in the generation of tongue and facial movements but is less important in the generation of jaw-closing movements. Our aim was to determine whether disruption of primate face motor cortical function would indeed interfere with the generation of tongue movements but would not interfere with the generation of jaw-closing movements. 2. The face motor cortex was reversibly inactivated with the use of cooling in two monkeys that were trained to perform both a tongue-protrusion task and a biting task. Recording of single neuronal activity in the cortex beneath the thermode confirmed the reversible inactivation of the cortex. Each task involved a series of trials in which the monkey was required to produce a preset force level for a 0.5-s force holding period; the monkey received a fruit-juice reward if it successfully completed a task trial. Cooling of the ICMS-defined face motor cortex was achieved bilaterally or, in one experiment, unilaterally by circulating coolant through thermodes placed either on intact dura overlying face motor cortex in both monkeys or directly on the exposed pia in one of the monkeys;thermode temperature was lowered to 3-5 degrees C during cooling. Electromyographic (EMG) recordings were also made from masseter, genioglossus, and digastric muscles. 3. During bilateral cooling of the thermodes on the dura overlying the face motor cortex, there was a significant reduction in the success rates for the performance of the tongue-protrusion task in comparison with control series of trials (i.e., precool and postcool) in which the thermodes were kept at 37 degrees C. Quantitative analyses of

  8. Intra-observer reliability for measuring first and second toe and metatarsal protrusion distance using palpation-based tests: a test-retest study

    PubMed Central

    2014-01-01

    Background Measurement of first and second metatarsal and toe protrusion is frequently used to explain foot problems using x-rays, osteological measurements or palpation-based tests. Length differences could be related to the appearance of problems in the foot. A test-retest design was conducted in order to establish the intra-rater reliability of three palpation-based tests. Methods 202 feet of physical therapy students and teachers of the CEU San Pablo University of Madrid, 39 men and 62 women, were measured using three different tests. Data were analysed using SPSS version 15.0. Mean, SD and 95% CI were calculated for each variable. A normal distribution of quantitative data was assessed using the Kolmogorov-Smirnov test. The test-retest intra-rater reliability was assessed using an Intraclass Correlation Coefficient (ICC). The Standard Error Mean (SEM) and the Minimal Detectable Change (MDC) were also obtained. Results All the ICC values showed a high degree of reliability (Test 1 = 0.97, Test 2 = 0.86 and Test 3 = 0.88) as did the SEM (Test 1 = 0.07, Test 2 = 0.10 and Test 3 = 0.11) and the MDC (Test 1 = 0.21, Test 2 = 0.30 and Test 3 = 0.31). Conclusions Reliability of measuring first and second metatarsal and toe protrusion using the three palpation-based tests showed a high degree of reliability. PMID:25729437

  9. The Threefold Protrusions of Adeno-Associated Virus Type 8 Are Involved in Cell Surface Targeting as Well as Postattachment Processing

    PubMed Central

    Raupp, Christina; Naumer, Matthias; Müller, Oliver J.; Gurda, Brittney L.; Agbandje-McKenna, Mavis

    2012-01-01

    Adeno-associated virus (AAV) has attracted considerable interest as a vector for gene therapy owing its lack of pathogenicity and the wealth of available serotypes with distinct tissue tropisms. One of the most promising isolates for vector development, based on its superior gene transfer efficiency to the liver in small animals compared to AAV type 2 (AAV2), is AAV8. Comparison of the in vivo gene transduction of rAAV2 and rAAV8 in mice showed that single amino acid exchanges in the 3-fold protrusions of AAV8 in the surface loops comprised of residues 581 to 584 and 589 to 592 to the corresponding amino acids of AAV2 and vice versa had a strong influence on transduction efficiency and tissue tropism. Surprisingly, not only did conversion of AAV8 to AAV2 cap sequences increase the transduction efficiency and change tissue tropism but so did the reciprocal conversion of AAV2 to AAV8. Insertion of new peptide motifs at position 590 in AAV8 also enabled retargeting of AAV8 capsids to specific tissues, suggesting that these sequences can interact with receptors on the cell surface. However, a neutralizing monoclonal antibody that binds to amino acids 588QQNTA592 of AAV8 does not prevent cell binding and virus uptake, indicating that this region is not necessary for receptor binding but rather that the antibody interferes with an essential step of postattachment processing in which the 3-fold protrusion is also involved. This study supports a multifunctional role of the 3-fold region of AAV capsids in the infection process. PMID:22718833

  10. Omniphobic Membrane for Robust Membrane Distillation

    SciTech Connect

    Lin, SH; Nejati, S; Boo, C; Hu, YX; Osuji, CO; Ehmelech, M

    2014-11-01

    In this work, we fabricate an omniphobic microporous membrane for membrane distillation (MD) by modifying a hydrophilic glass fiber membrane with silica nanoparticles followed by surface fluorination and polymer coating. The modified glass fiber membrane exhibits an anti-wetting property not only against water but also against low surface tension organic solvents that easily wet a hydrophobic polytetrafluoroethylene (PTFE) membrane that is commonly used in MD applications. By comparing the performance of the PTFE and omniphobic membranes in direct contact MD experiments in the presence of a surfactant (sodium dodecyl sulfate, SDS), we show that SDS wets the hydrophobic PTFE membrane but not the omniphobic membrane. Our results suggest that omniphobic membranes are critical for MD applications with feed waters containing surface active species, such as oil and gas produced water, to prevent membrane pore wetting.

  11. Geometry of membrane fission.

    PubMed

    Frolov, Vadim A; Escalada, Artur; Akimov, Sergey A; Shnyrova, Anna V

    2015-01-01

    Cellular membranes define the functional geometry of intracellular space. Formation of new membrane compartments and maintenance of complex organelles require division and disconnection of cellular membranes, a process termed membrane fission. Peripheral membrane proteins generally control membrane remodeling during fission. Local membrane stresses, reflecting molecular geometry of membrane-interacting parts of these proteins, sum up to produce the key membrane geometries of fission: the saddle-shaped neck and hour-glass hemifission intermediate. Here, we review the fundamental principles behind the translation of molecular geometry into membrane shape and topology during fission. We emphasize the central role the membrane insertion of specialized protein domains plays in orchestrating fission in vitro and in cells. We further compare individual to synergistic action of the membrane insertion during fission mediated by individual protein species, proteins complexes or membrane domains. Finally, we describe how local geometry of fission intermediates defines the functional design of the protein complexes catalyzing fission of cellular membranes. PMID:25062896

  12. Self-organized antireflection CuIn(S,Se)2 nano-protrusions on flexible substrates by ion erosion based on CuInS2 nanocrystal precursor inks

    NASA Astrophysics Data System (ADS)

    Yen, Yu-Ting; Wang, Yi-Chung; Chen, Chia-Wei; Tsai, Hung-Wei; Chen, Yu-Ze; Hu, Fan; Chueh, Yu-Lun

    2015-11-01

    In this work, an approach to achieve surface nano-protrusions on a chalcopyrite CuIn(S,Se)2 thin film was demonstrated. Home-made CuInS2 nanocrystals with average diameter of 20 nm were prepared and characterized. By applying ion erosion process on the CuIn(S,Se)2 film, large-area self-aligned nano-protrusions can be formed. Interestingly, the process can be applied on flexible substrate where the CuIn(S,Se)2 film remains intact with no visible cracking after several bending tests. In addition, reflectance spectra reveal the extraordinary anti-reflectance characteristics of nano-protrusions on the CuIn(S,Se)2 film with the incident light from 350 to 2000 nm. A 36-cm2 CuIn(S,Se)2 film with nano-protrusions on flexible molybdenum foil substrate has been demonstrated, which demonstrated the feasibility of developing low cost with a high optical absorption CuIn(S,Se)2 flexible thin film.

  13. Anion permselective membrane

    NASA Technical Reports Server (NTRS)

    Hodgdon, R. B.; Waite, W. A.

    1980-01-01

    The efforts on the synthesis of polymer anion redox membranes were mainly concentrated in two areas, membrane development and membrane fabrication. Membrane development covered the preparation and evaluation of experimental membranes systems with improved resistance stability and/or lower permeability. Membrane fabrication covered the laboratory scale production of prime candidate membranes in quantities of up to two hundred and sizes up to 18 inches x 18 inches (46 cm x 46 cm). These small (10 in x 11 in) and medium sized membranes were mainly for assembly into multicell units. Improvements in processing procedures and techniques for preparing such membrane sets lifted yields to over 90 percent.

  14. Selecting a Roof Membrane.

    ERIC Educational Resources Information Center

    Waldron, Larry W.

    1990-01-01

    Offers a brief synopsis of the unique characteristics of the following roof membranes: (1) built-up roofing; (2) elastoplastic membranes; (3) modified bitumen membranes; (4) liquid applied membranes; and (5) metal roofing. A chart compares the characteristics of the raw membranes only. (MLF)

  15. Membrane Systems in Cyanobacteria

    SciTech Connect

    Liberton, Michelle L.; Pakrasi, Himadri B.

    2008-01-01

    Cyanobacteria are photosynthetic prokaryotes with highly differentiated membrane systems. In addition to a Gram-negative-type cell envelope with plasma membrane and outer membrane separated by a periplasmic space, cyanobacteria have an internal system of thylakoid membranes where the fully functional electron transfer chains of photosynthesis and respiration reside. The presence of different membrane systems lends these cells a unique complexity among bacteria. Cyanobacteria must be able to reorganize the membranes, synthesize new membrane lipids, and properly target proteins to the correct membrane system. The outer membrane, plasma membrane, and thylakoid membranes each have specialized roles in the cyanobacterial cell. Understanding the organization, functionality, protein composition and dynamics of the membrane systems remains a great challenge in cyanobacterial cell biology.

  16. Three-dimensional organization of nascent rod outer segment disk membranes

    PubMed Central

    Volland, Stefanie; Hughes, Louise C.; Kong, Christina; Burgess, Barry L.; Linberg, Kenneth A.; Luna, Gabriel; Zhou, Z. Hong; Fisher, Steven K.; Williams, David S.

    2015-01-01

    The vertebrate photoreceptor cell contains an elaborate cilium that includes a stack of phototransductive membrane disks. The disk membranes are continually renewed, but how new disks are formed remains poorly understood. Here we used electron microscope tomography to obtain 3D visualization of the nascent disks of rod photoreceptors in three mammalian species, to gain insight into the process of disk morphogenesis. We observed that nascent disks are invariably continuous with the ciliary plasma membrane, although, owing to partial enclosure, they can appear to be internal in 2D profiles. Tomographic analyses of the basal-most region of the outer segment show changes in shape of the ciliary plasma membrane indicating an invagination, which is likely a first step in disk formation. The invagination flattens to create the proximal surface of an evaginating lamella, as well as membrane protrusions that extend between adjacent lamellae, thereby initiating a disk rim. Immediately distal to this initiation site, lamellae of increasing diameter are evident, indicating growth outward from the cilium. In agreement with a previous model, our data indicate that mature disks are formed once lamellae reach full diameter, and the growth of a rim encloses the space between adjacent surfaces of two lamellae. This study provides 3D data of nascent and mature rod photoreceptor disk membranes at unprecedented z-axis depth and resolution, and provides a basis for addressing fundamental questions, ranging from protein sorting in the photoreceptor cilium to photoreceptor electrophysiology. PMID:26578801

  17. Disentangling Membrane Dynamics and Cell Migration; Differential Influences of F-actin and Cell-Matrix Adhesions.

    PubMed

    Kowalewski, Jacob M; Shafqat-Abbasi, Hamdah; Jafari-Mamaghani, Mehrdad; Endrias Ganebo, Bereket; Gong, Xiaowei; Strömblad, Staffan; Lock, John G

    2015-01-01

    Cell migration is heavily interconnected with plasma membrane protrusion and retraction (collectively termed "membrane dynamics"). This makes it difficult to distinguish regulatory mechanisms that differentially influence migration and membrane dynamics. Yet such distinctions may be valuable given evidence that cancer cell invasion in 3D may be better predicted by 2D membrane dynamics than by 2D cell migration, implying a degree of functional independence between these processes. Here, we applied multi-scale single cell imaging and a systematic statistical approach to disentangle regulatory associations underlying either migration or membrane dynamics. This revealed preferential correlations between membrane dynamics and F-actin features, contrasting with an enrichment of links between cell migration and adhesion complex properties. These correlative linkages were often non-linear and therefore context-dependent, strengthening or weakening with spontaneous heterogeneity in cell behavior. More broadly, we observed that slow moving cells tend to increase in area, while fast moving cells tend to shrink, and that the size of dynamic membrane domains is independent of cell area. Overall, we define macromolecular features preferentially associated with either cell migration or membrane dynamics, enabling more specific interrogation and targeting of these processes in future.

  18. Chloroplast protrusions in leaves of Ranunculus glacialis L. respond significantly to different ambient conditions, but are not related to temperature stress.

    PubMed

    Moser, Tim; Holzinger, Andreas; Buchner, Othmar

    2015-07-01

    The occurrence of chloroplast protrusions (CPs) in leaves of Ranunculus glacialis L. in response to different environmental conditions was assessed. CPs occur highly dynamically. They do not contain thylakoids and their physiological function is still largely unknown. Controlled in situ sampling showed that CP formation follows a pronounced diurnal rhythm. Between 2 and 27 °C the relative proportion of chloroplasts with CPs (rCP) showed a significant positive correlation to leaf temperature (TL; 0.793, P < 0.01), while irradiation intensity had a minor effect on rCP. In situ shading and controlled laboratory experiments confirmed the significant influence of TL. Under moderate irradiation intensity, an increase of TL up to 25 °C significantly promoted CP formation, while a further increase to 37 °C led to a decrease. Furthermore, rCP values were lower in darkness and under high irradiation intensity. Gas treatment at 2000 ppm CO2/2% O2 led to a significant decrease of rCP, suggesting a possible involvement of photorespiration in CP formation. Our findings demonstrate that in R. glacialis, CPs are neither a rare phenomenon nor a result of heat or light stress; on the contrary, they seem to be most abundant under moderate temperature and non-stress irradiation conditions.

  19. Composite sensor membrane

    SciTech Connect

    Majumdar, Arun; Satyanarayana, Srinath; Yue, Min

    2008-03-18

    A sensor may include a membrane to deflect in response to a change in surface stress, where a layer on the membrane is to couple one or more probe molecules with the membrane. The membrane may deflect when a target molecule reacts with one or more probe molecules.

  20. Experimenting with Liquid Membranes.

    ERIC Educational Resources Information Center

    Lamb, J. D.; And Others

    1980-01-01

    Outlined are two experiments using liquid membranes that illustrate carrier-facilitated transport, where chemical species are ushered across the membrane by selective "carrier" molecules residing in the membrane. The use of liquid membranes as models for studying and describing biological transport mechanisms is explored. (CS)

  1. Electrostatically shaped membranes

    NASA Technical Reports Server (NTRS)

    Silverberg, Larry M. (Inventor)

    1994-01-01

    Disclosed is a method and apparatus for electrostatically shaping a membrane suitable for use in antennas or the like, comprising an electrically conductive thin membrane where the periphery of said membrane is free to move in at least one direction, a first charge on the electrically conductive thin membrane to electrostatically stiffen the membrane, a second charge which shapes the electrostatically stiffened thin membrane and a restraint for limiting the movement of at least one point of the thin membrane relative to the second charge. Also disclosed is a method and apparatus for adaptively controlling the shape of the thin membrane by sensing the shape of the membrane and selectively controlling the first and second charge to achieve a desired performance characteristic of the membrane.

  2. Outer-Inner Membrane Vesicles Naturally Secreted by Gram-Negative Pathogenic Bacteria

    PubMed Central

    Pérez-Cruz, Carla; Delgado, Lidia; López-Iglesias, Carmen; Mercade, Elena

    2015-01-01

    Outer-inner membrane vesicles (O-IMVs) were recently described as a new type of membrane vesicle secreted by the Antarctic bacterium Shewanella vesiculosa M7T. Their formation is characterized by the protrusion of both outer and plasma membranes, which pulls cytoplasmic components into the vesicles. To demonstrate that this is not a singular phenomenon in a bacterium occurring in an extreme environment, the identification of O-IMVs in pathogenic bacteria was undertaken. With this aim, a structural study by Transmission Electron Microscopy (TEM) and Cryo-transmission electron microscopy (Cryo-TEM) was carried out, confirming that O-IMVs are also secreted by Gram-negative pathogenic bacteria such as Neisseria gonorrhoeae, Pseudomonas aeruginosa PAO1 and Acinetobacter baumannii AB41, in which they represent between 0.23% and 1.2% of total vesicles produced. DNA and ATP, which are components solely found in the cell cytoplasm, were identified within membrane vesicles of these strains. The presence of DNA inside the O-IMVs produced by N. gonorrhoeae was confirmed by gold DNA immunolabeling with a specific monoclonal IgM against double-stranded DNA. A proteomic analysis of N. gonorrhoeae-derived membrane vesicles identified proteins from the cytoplasm and plasma membrane. This confirmation of O-IMV extends the hitherto uniform definition of membrane vesicles in Gram-negative bacteria and explains the presence of components in membrane vesicles such as DNA, cytoplasmic and inner membrane proteins, as well as ATP, detected for the first time. The production of these O-IMVs by pathogenic Gram-negative bacteria opens up new areas of study related to their involvement in lateral gene transfer, the transfer of cytoplasmic proteins, as well as the functionality and role of ATP detected in these new vesicles. PMID:25581302

  3. Membrane position control

    NASA Technical Reports Server (NTRS)

    Su, Ji (Inventor); Harrison, Joycelyn S. (Inventor)

    2004-01-01

    A membrane structure includes at least one electroactive bending actuator fixed to a supporting base. Each electroactive bending actuator is operatively connected to the membrane for controlling membrane position. Any displacement of each electroactive bending actuator effects displacement of the membrane. More specifically, the operative connection is provided by a guiding wheel assembly and a track, wherein displacement of the bending actuator effects translation of the wheel assembly along the track, thereby imparting movement to the membrane.

  4. Immunohistochemical localization of glucose transporters and insulin receptors in human fetal membranes at term.

    PubMed

    Wolf, H J; Desoye, G

    1993-11-01

    The localization has been investigated of the isoforms GLUT1, GLUT3 and GLUT4 of glucose transporter proteins as well as of insulin receptors. Fetal membranes (n = 10) were examined by immunohistochemical methods at the light and electron microscopic levels using mono- and polyclonal antibodies. In all amnion epithelial cells, GLUT1 and GLUT3 antibodies were bound to the apical membrane. Very rarely the GLUT1 antibody also immunostained the basolateral membrane and reacted weakly with the endomembrane system and membranes of the lateral cell protrusions. Fibroblasts reacted with the antibodies against GLUT1, GLUT4 and insulin receptor, whereas they were labelled only in one case with GLUT3 antibody. Cytotrophoblast cells were only stained with antibodies against GLUT1 and GLUT3. Antibodies against GLUT4 only reacted with fibroblasts in the membranes. On amnion epithelial cells, weak immunoreactivity with insulin receptor antibodies was detected only at the electron microscopic level. The data indicate: (1) GLUT1 is located on all cells of the amnion, whereas GLUT3 is present in detectable amounts only on amnion epithelial cells and cytotrophoblast; (2) GLUT1 and GLUT3 on amnion epithelial cells are predominantly located on the apical surface; (3) GLUT4 and insulin receptors are not regularly expressed. We suggest that amnion epithelial cells cover their basal glucose requirements from the amniotic fluid and not from the maternal circulation.

  5. Membrane shrinkage and cortex remodelling are predicted to work in harmony to retract blebs.

    PubMed

    Woolley, Thomas E; Gaffney, Eamonn A; Goriely, Alain

    2015-07-01

    Numerous cell types undergo an oscillatory form of dynamics known as blebbing, whereby pressure-driven spherical protrusions of membrane (known as blebs) expand and contract over the cell's surface. Depending on the cell line, blebs play important roles in many different phenomena including mitosis and locomotion. The expansion phase of cellular blebbing has been mathematically modelled in detail. However, the active processes occurring during the retraction phase are not so well characterized. It is thought that blebs retract because a cortex reforms inside, and adheres to, the bleb membrane. This cortex is retracted into the cell and the attached bleb membrane follows. Using a computational model of a cell's membrane, cortex and interconnecting adhesions, we demonstrate that cortex retraction alone cannot account for bleb retraction and suggest that the mechanism works in tandem with membrane shrinking. Further, an emergent hysteresis loop is observed in the intracellular pressure, which suggests a potential mechanism through which a secondary bleb can be initiated as a primary bleb contracts. PMID:26587278

  6. Membrane tubule formation by banana-shaped proteins with or without transient network structure

    NASA Astrophysics Data System (ADS)

    Noguchi, Hiroshi

    2016-02-01

    In living cells, membrane morphology is regulated by various proteins. Many membrane reshaping proteins contain a Bin/Amphiphysin/Rvs (BAR) domain, which consists of a banana-shaped rod. The BAR domain bends the biomembrane along the rod axis and the features of this anisotropic bending have recently been studied. Here, we report on the role of the BAR protein rods in inducing membrane tubulation, using large-scale coarse-grained simulations. We reveal that a small spontaneous side curvature perpendicular to the rod can drastically alter the tubulation dynamics at high protein density, whereas no significant difference is obtained at low density. A percolated network is intermediately formed depending on the side curvature. This network suppresses tubule protrusion, leading to the slow formation of fewer tubules. Thus, the side curvature, which is generated by protein-protein and membrane-protein interactions, plays a significant role in tubulation dynamics. We also find that positive surface tensions and the vesicle membrane curvature can stabilize this network structure by suppressing the tubulation.

  7. Membrane shrinkage and cortex remodelling are predicted to work in harmony to retract blebs

    PubMed Central

    Woolley, Thomas E.; Gaffney, Eamonn A.; Goriely, Alain

    2015-01-01

    Numerous cell types undergo an oscillatory form of dynamics known as blebbing, whereby pressure-driven spherical protrusions of membrane (known as blebs) expand and contract over the cell's surface. Depending on the cell line, blebs play important roles in many different phenomena including mitosis and locomotion. The expansion phase of cellular blebbing has been mathematically modelled in detail. However, the active processes occurring during the retraction phase are not so well characterized. It is thought that blebs retract because a cortex reforms inside, and adheres to, the bleb membrane. This cortex is retracted into the cell and the attached bleb membrane follows. Using a computational model of a cell's membrane, cortex and interconnecting adhesions, we demonstrate that cortex retraction alone cannot account for bleb retraction and suggest that the mechanism works in tandem with membrane shrinking. Further, an emergent hysteresis loop is observed in the intracellular pressure, which suggests a potential mechanism through which a secondary bleb can be initiated as a primary bleb contracts. PMID:26587278

  8. Re-analysis of protein data reveals the germination pathway and up accumulation mechanism of cell wall hydrolases during the radicle protrusion step of seed germination in Podophyllum hexandrum- a high altitude plant.

    PubMed

    Dogra, Vivek; Bagler, Ganesh; Sreenivasulu, Yelam

    2015-01-01

    Podophyllum hexandrum Royle is an important high-altitude plant of Himalayas with immense medicinal value. Earlier, it was reported that the cell wall hydrolases were up accumulated during radicle protrusion step of Podophyllum seed germination. In the present study, Podophyllum seed Germination protein interaction Network (PGN) was constructed by using the differentially accumulated protein (DAP) data set of Podophyllum during the radicle protrusion step of seed germination, with reference to Arabidopsis protein-protein interaction network (AtPIN). The developed PGN is comprised of a giant cluster with 1028 proteins having 10,519 interactions and a few small clusters with relevant gene ontological signatures. In this analysis, a germination pathway related cluster which is also central to the topology and information dynamics of PGN was obtained with a set of 60 key proteins. Among these, eight proteins which are known to be involved in signaling, metabolism, protein modification, cell wall modification, and cell cycle regulation processes were found commonly highlighted in both the proteomic and interactome analysis. The systems-level analysis of PGN identified the key proteins involved in radicle protrusion step of seed germination in Podophyllum.

  9. Re-analysis of protein data reveals the germination pathway and up accumulation mechanism of cell wall hydrolases during the radicle protrusion step of seed germination in Podophyllum hexandrum- a high altitude plant

    PubMed Central

    Dogra, Vivek; Bagler, Ganesh; Sreenivasulu, Yelam

    2015-01-01

    Podophyllum hexandrum Royle is an important high-altitude plant of Himalayas with immense medicinal value. Earlier, it was reported that the cell wall hydrolases were up accumulated during radicle protrusion step of Podophyllum seed germination. In the present study, Podophyllum seed Germination protein interaction Network (PGN) was constructed by using the differentially accumulated protein (DAP) data set of Podophyllum during the radicle protrusion step of seed germination, with reference to Arabidopsis protein–protein interaction network (AtPIN). The developed PGN is comprised of a giant cluster with 1028 proteins having 10,519 interactions and a few small clusters with relevant gene ontological signatures. In this analysis, a germination pathway related cluster which is also central to the topology and information dynamics of PGN was obtained with a set of 60 key proteins. Among these, eight proteins which are known to be involved in signaling, metabolism, protein modification, cell wall modification, and cell cycle regulation processes were found commonly highlighted in both the proteomic and interactome analysis. The systems-level analysis of PGN identified the key proteins involved in radicle protrusion step of seed germination in Podophyllum. PMID:26579141

  10. Src1 is a Protein of the Inner Nuclear Membrane Interacting with the Dictyostelium Lamin NE81

    PubMed Central

    Batsios, Petros; Ren, Xiang; Baumann, Otto; Larochelle, Denis A.; Gräf, Ralph

    2016-01-01

    The nuclear envelope (NE) consists of the outer and inner nuclear membrane (INM), whereby the latter is bound to the nuclear lamina. Src1 is a Dictyostelium homologue of the helix-extension-helix family of proteins, which also includes the human lamin-binding protein MAN1. Both endogenous Src1 and GFP-Src1 are localized to the NE during the entire cell cycle. Immuno-electron microscopy and light microscopy after differential detergent treatment indicated that Src1 resides in the INM. FRAP experiments with GFP-Src1 cells suggested that at least a fraction of the protein could be stably engaged in forming the nuclear lamina together with the Dictyostelium lamin NE81. Both a BioID proximity assay and mis-localization of soluble, truncated mRFP-Src1 at cytosolic clusters consisting of an intentionally mis-localized mutant of GFP-NE81 confirmed an interaction of Src1 and NE81. Expression GFP-Src11–646, a fragment C-terminally truncated after the first transmembrane domain, disrupted interaction of nuclear membranes with the nuclear lamina, as cells formed protrusions of the NE that were dependent on cytoskeletal pulling forces. Protrusions were dependent on intact microtubules but not actin filaments. Our results indicate that Src1 is required for integrity of the NE and highlight Dictyostelium as a promising model for the evolution of nuclear architecture. PMID:26999214

  11. Src1 is a Protein of the Inner Nuclear Membrane Interacting with the Dictyostelium Lamin NE81.

    PubMed

    Batsios, Petros; Ren, Xiang; Baumann, Otto; Larochelle, Denis A; Gräf, Ralph

    2016-03-18

    The nuclear envelope (NE) consists of the outer and inner nuclear membrane (INM), whereby the latter is bound to the nuclear lamina. Src1 is a Dictyostelium homologue of the helix-extension-helix family of proteins, which also includes the human lamin-binding protein MAN1. Both endogenous Src1 and GFP-Src1 are localized to the NE during the entire cell cycle. Immuno-electron microscopy and light microscopy after differential detergent treatment indicated that Src1 resides in the INM. FRAP experiments with GFP-Src1 cells suggested that at least a fraction of the protein could be stably engaged in forming the nuclear lamina together with the Dictyostelium lamin NE81. Both a BioID proximity assay and mis-localization of soluble, truncated mRFP-Src1 at cytosolic clusters consisting of an intentionally mis-localized mutant of GFP-NE81 confirmed an interaction of Src1 and NE81. Expression GFP-Src1(1-646), a fragment C-terminally truncated after the first transmembrane domain, disrupted interaction of nuclear membranes with the nuclear lamina, as cells formed protrusions of the NE that were dependent on cytoskeletal pulling forces. Protrusions were dependent on intact microtubules but not actin filaments. Our results indicate that Src1 is required for integrity of the NE and highlight Dictyostelium as a promising model for the evolution of nuclear architecture.

  12. Correlation of cell membrane dynamics and cell motility

    PubMed Central

    2011-01-01

    Background Essential events of cell development and homeostasis are revealed by the associated changes of cell morphology and therefore have been widely used as a key indicator of physiological states and molecular pathways affecting various cellular functions via cytoskeleton. Cell motility is a complex phenomenon primarily driven by the actin network, which plays an important role in shaping the morphology of the cells. Most of the morphology based features are approximated from cell periphery but its dynamics have received none to scant attention. We aim to bridge the gap between membrane dynamics and cell states from the perspective of whole cell movement by identifying cell edge patterns and its correlation with cell dynamics. Results We present a systematic study to extract, classify, and compare cell dynamics in terms of cell motility and edge activity. Cell motility features extracted by fitting a persistent random walk were used to identify the initial set of cell subpopulations. We propose algorithms to extract edge features along the entire cell periphery such as protrusion and retraction velocity. These constitute a unique set of multivariate time-lapse edge features that are then used to profile subclasses of cell dynamics by unsupervised clustering. Conclusions By comparing membrane dynamic patterns exhibited by each subclass of cells, correlated trends of edge and cell movements were identified. Our findings are consistent with published literature and we also identified that motility patterns are influenced by edge features from initial time points compared to later sampling intervals. PMID:22372978

  13. Correlative Light and Electron Microscopy Reveals the HAS3-Induced Dorsal Plasma Membrane Ruffles

    PubMed Central

    Rilla, Kirsi; Koistinen, Arto

    2015-01-01

    Hyaluronan is a linear sugar polymer synthesized by three isoforms of hyaluronan synthases (HAS1, 2, and 3) that forms a hydrated scaffold around cells and is an essential component of the extracellular matrix. The morphological changes of cells induced by active hyaluronan synthesis are well recognized but not studied in detail with high resolution before. We have previously found that overexpression of HAS3 induces growth of long plasma membrane protrusions that act as platforms for hyaluronan synthesis. The study of these thin and fragile protrusions is challenging, and they are difficult to preserve by fixation unless they are adherent to the substrate. Thus their structure and regulation are still partly unclear despite careful imaging with different microscopic methods in several cell types. In this study, correlative light and electron microscopy (CLEM) was utilized to correlate the GFP-HAS3 signal and the surface ultrastructure of cells in order to study in detail the morphological changes induced by HAS3 overexpression. Surprisingly, this method revealed that GFP-HAS3 not only localizes to ruffles but in fact induces dorsal ruffle formation. Dorsal ruffles regulate diverse cellular functions, such as motility, regulation of glucose metabolism, spreading, adhesion, and matrix degradation, the same functions driven by active hyaluronan synthesis. PMID:26448759

  14. Intersectin adaptor proteins are associated with actin-regulating protein WIP in invadopodia.

    PubMed

    Gryaznova, Tetyana; Kropyvko, Sergii; Burdyniuk, Mariia; Gubar, Olga; Kryklyva, Valentyna; Tsyba, Liudmyla; Rynditch, Alla

    2015-07-01

    Invasive cancer cells form actin-rich membrane protrusions called invadopodia that degrade extracellular matrix and facilitate cell invasion and metastasis. WIP (WASP-interacting protein) together with N-WASP (neural Wiskott-Aldrich syndrome protein) are localized in invadopodia and play a crucial role in their formation. Here we show that WIP interacts with endocytic adaptor proteins of the intersectin (ITSN) family, ITSN1 and ITSN2. The interaction is mediated by the SH3 domains of ITSNs and the middle part of the WIP proline-rich motifs. We have also demonstrated that ITSN1, WIP and N-WASP can form a complex in cells. Endogenous ITSN1 and ITSN2 are located in invasive protrusions of MDA-MB-231 breast cancer cell line. Moreover, data from immunofluorescent analysis revealed co-localization of ITSN1 and WIP at sites of invadopodia formation and in clathrin-coated pits. Together, these findings provide insights into the molecular mechanisms of invadopodia formation and identify ITSNs as scaffold proteins involved in this process.

  15. Sheet Membrane Spacesuit Water Membrane Evaporator

    NASA Technical Reports Server (NTRS)

    Bue, Grant; Trevino, Luis; Zapata, Felipe; Dillion, Paul; Castillo, Juan; Vonau, Walter; Wilkes, Robert; Vogel, Matthew; Frodge, Curtis

    2013-01-01

    A document describes a sheet membrane spacesuit water membrane evaporator (SWME), which allows for the use of one common water tank that can supply cooling water to the astronaut and to the evaporator. Test data showed that heat rejection performance dropped only 6 percent after being subjected to highly contaminated water. It also exhibited robustness with respect to freezing and Martian atmospheric simulation testing. Water was allowed to freeze in the water channels during testing that simulated a water loop failure and vapor backpressure valve failure. Upon closing the backpressure valve and energizing the pump, the ice eventually thawed and water began to flow with no apparent damage to the sheet membrane. The membrane evaporator also serves to de-gas the water loop from entrained gases, thereby eliminating the need for special degassing equipment such as is needed by the current spacesuit system. As water flows through the three annular water channels, water evaporates with the vapor flowing across the hydrophobic, porous sheet membrane to the vacuum side of the membrane. The rate at which water evaporates, and therefore, the rate at which the flowing water is cooled, is a function of the difference between the water saturation pressure on the water side of the membrane, and the pressure on the vacuum side of the membrane. The primary theory is that the hydrophobic sheet membrane retains water, but permits vapor pass-through when the vapor side pressure is less than the water saturation pressure. This results in evaporative cooling of the remaining water.

  16. Membrane selectivity in pervaporation

    SciTech Connect

    Kujawski, W.

    1996-06-01

    A qualitative description is presented of pervaporation which discusses the initial preferential sorption into the membrane, diffusion of liquid, phase transition from liquid to vapor phase, followed by diffusion of vapors and fast desorption from the other side of the membrane. The overall separation of each pervaporation step was calculated in terms of separation factor {alpha}. The results show that in the case of hydrophilic membranes (i.e., dense polyamide-6 membrane and ion-exchange membrane PESS-1) and water-ethanol mixtures, the phase transition step decreases the overall separation. Also, diffusion through the membrane is unfavorable to water at a low concentration range.

  17. Imaging excised apical plasma membrane patches of MDCK cells in physiological conditions with atomic force microscopy.

    PubMed

    Lärmer, J; Schneider, S W; Danker, T; Schwab, A; Oberleithner, H

    1997-07-01

    We combined the patch-clamp technique with atomic force microscopy (AFM) to visualize plasma membrane proteins protruding from the extracellular surface of cultured kidney cells (MDCK cells). To achieve molecular resolution, patches were mechanically isolated from whole MDCK cells by applying the patch-clamp technique. The excised inside-out patches were transferred on freshly cleaved mica and imaged with the AFM in air and under physiological conditions (i. e. in fluid). Thus, the resolution could be increased considerably (lateral and vertical resolutions 5 and 0.1 nm, respectively) as compared to experiments on intact cells, where plasma membrane proteins were hardly detectable. The apical plasma membrane surface of the MDCK cells showed multiple protrusions which could be identified as membrane proteins through the use of pronase. These proteins had a density of about 90 per micron(2), with heights between 1 and 9 nm, and lateral dimensions of 20-60 nm. Their frequency distribution showed a peak value of 3 nm for the protein height. A simplified assumption - modelling plasma membrane proteins as spherical structures protruding from the lipid bilayer - allowed an estimation of the possible molecular weights of these proteins. They range from 50 kDa to 710 kDa with a peak value of 125 kDa. We conclude that AFM can be used to study the molecular structures of membranes which were isolated with the patch-clamp technique. Individual membrane proteins and protein clusters, and their arrangement and distribution in a native plasma membrane can be visualized under physiological conditions, which is a first step for their identification. PMID:9178623

  18. Hybrid adsorptive membrane reactor

    NASA Technical Reports Server (NTRS)

    Tsotsis, Theodore T. (Inventor); Sahimi, Muhammad (Inventor); Fayyaz-Najafi, Babak (Inventor); Harale, Aadesh (Inventor); Park, Byoung-Gi (Inventor); Liu, Paul K. T. (Inventor)

    2011-01-01

    A hybrid adsorbent-membrane reactor in which the chemical reaction, membrane separation, and product adsorption are coupled. Also disclosed are a dual-reactor apparatus and a process using the reactor or the apparatus.

  19. Composite zeolite membranes

    DOEpatents

    Nenoff, Tina M.; Thoma, Steven G.; Ashley, Carol S.; Reed, Scott T.

    2002-01-01

    A new class of composite zeolite membranes and synthesis techniques therefor has been invented. These membranes are essentially defect-free, and exhibit large levels of transmembrane flux and of chemical and isotopic selectivity.

  20. Supported inorganic membranes

    DOEpatents

    Sehgal, Rakesh; Brinker, Charles Jeffrey

    1998-01-01

    Supported inorganic membranes capable of molecular sieving, and methods for their production, are provided. The subject membranes exhibit high flux and high selectivity. The subject membranes are substantially defect free and less than about 100 nm thick. The pores of the subject membranes have an average critical pore radius of less than about 5 .ANG., and have a narrow pore size distribution. The subject membranes are prepared by coating a porous substrate with a polymeric sol, preferably under conditions of low relative pressure of the liquid constituents of the sol. The coated substrate is dried and calcined to produce the subject supported membrane. Also provided are methods of derivatizing the surface of supported inorganic membranes with metal alkoxides. The subject membranes find use in a variety of applications, such as the separation of constituents of gaseous streams, as catalysts and catalyst supports, and the like.

  1. Tympanic membrane (image)

    MedlinePlus

    ... tympanic membrane they cause it to vibrate. The vibrations are then transferred to the tiny bones in the middle ear. The middle ear bones then transfer the vibrating signals to the inner ear. The tympanic membrane is ...

  2. Hybrid adsorptive membrane reactor

    DOEpatents

    Tsotsis, Theodore T.; Sahimi, Muhammad; Fayyaz-Najafi, Babak; Harale, Aadesh; Park, Byoung-Gi; Liu, Paul K. T.

    2011-03-01

    A hybrid adsorbent-membrane reactor in which the chemical reaction, membrane separation, and product adsorption are coupled. Also disclosed are a dual-reactor apparatus and a process using the reactor or the apparatus.

  3. Composite fuel cell membranes

    DOEpatents

    Plowman, Keith R.; Rehg, Timothy J.; Davis, Larry W.; Carl, William P.; Cisar, Alan J.; Eastland, Charles S.

    1997-01-01

    A bilayer or trilayer composite ion exchange membrane suitable for use in a fuel cell. The composite membrane has a high equivalent weight thick layer in order to provide sufficient strength and low equivalent weight surface layers for improved electrical performance in a fuel cell. In use, the composite membrane is provided with electrode surface layers. The composite membrane can be composed of a sulfonic fluoropolymer in both core and surface layers.

  4. Composite fuel cell membranes

    DOEpatents

    Plowman, K.R.; Rehg, T.J.; Davis, L.W.; Carl, W.P.; Cisar, A.J.; Eastland, C.S.

    1997-08-05

    A bilayer or trilayer composite ion exchange membrane is described suitable for use in a fuel cell. The composite membrane has a high equivalent weight thick layer in order to provide sufficient strength and low equivalent weight surface layers for improved electrical performance in a fuel cell. In use, the composite membrane is provided with electrode surface layers. The composite membrane can be composed of a sulfonic fluoropolymer in both core and surface layers.

  5. Industrial membrane processes

    SciTech Connect

    White, R.E.; Pintauro, P.N.

    1986-01-01

    This book presents the papers given a symposium on the use of membranes in industrial plants. Topics considered include the effect of biofilm formation on salinity power plant output on a laboratory scale, an engineering analysis of membrane-aided distillation, the development and evaluation of sulfonated polysulfone membranes for the zinc/ferricyanide battery, and the degradation of ionic membranes in the zinc/ferricyanide battery.

  6. Release of Membrane-Bound Vesicles and Inhibition of Tumor Cell Adhesion by the Peptide Neopetrosiamide A

    PubMed Central

    Austin, Pamela; Heller, Markus; Williams, David E.; McIntosh, Lawrence P.; Vogl, A. Wayne; Foster, Leonard J.; Andersen, Raymond J.; Roberge, Michel; Roskelley, Calvin D.

    2010-01-01

    Background Neopetrosiamide A (NeoA) is a 28-amino acid tricyclic peptide originally isolated from a marine sponge as a tumor cell invasion inhibitor whose mechanism of action is unknown. Methodology/Principal Findings We show that NeoA reversibly inhibits tumor cell adhesion, disassembles focal adhesions in pre-attached cells, and decreases the level of β1 integrin subunits on the cell surface. NeoA also induces the formation of dynamic, membrane-bound protrusions on the surface of treated cells and the release of membrane-bound vesicles into the culture medium. Proteomic analysis indicates that the vesicles contain EGF and transferrin receptors as well as a number of proteins involved in adhesion and migration including: β1 integrin and numerous α integrin subunits; actin and actin-binding proteins such as cofilin, moesin and myosin 1C; and membrane modulating eps15 homology domain (EHD) proteins. Surface labeling, trafficking inhibition, and real-time imaging experiments all suggest that β1 integrin-containing vesicles are released directly from NeoA-induced cell surface protrusions rather than from vesicles generated intracellularly. The biological activity of NeoA is dependent on its disulfide bond pattern and NMR spectroscopy indicates that the peptide is globular with a continuous ridge of hydrophobic groups flanked by charged amino acid residues that could facilitate a simultaneous interaction with lipids and proteins in the membrane. Conclusions/Significance NeoA is an anti-adhesive peptide that decreases cell surface integrin levels through a novel, yet to be elucidated, mechanism that involves the release of adhesion molecule-containing vesicles from the cell surface. PMID:20520768

  7. Cadmium sulfide membranes

    DOEpatents

    Spanhel, Lubomir; Anderson, Marc A.

    1992-07-07

    A method is described for the creation of novel q-effect cadmium sulfide membranes. The membranes are made by first creating a dilute cadmium sulfide colloid in aqueous suspension and then removing the water and excess salts therefrom. The cadmium sulfide membrane thus produced is luminescent at room temperature and may have application in laser fabrication.

  8. Cadmium sulfide membranes

    DOEpatents

    Spanhel, Lubomir; Anderson, Marc A.

    1991-10-22

    A method is described for the creation of novel q-effect cadmium sulfide membranes. The membranes are made by first creating a dilute cadmium sulfide colloid in aqueous suspension and then removing the water and excess salts therefrom. The cadmium sulfide membrane thus produced is luminescent at room temperature and may have application in laser fabrication.

  9. Meniscus Membranes For Separation

    DOEpatents

    Dye, Robert C.; Jorgensen, Betty; Pesiri, David R.

    2005-09-20

    Gas separation membranes, especially meniscus-shaped membranes for gas separations are disclosed together with the use of such meniscus-shaped membranes for applications such as thermal gas valves, pre-concentration of a gas stream, and selective pre-screening of a gas stream. In addition, a rapid screening system for simultaneously screening polymer materials for effectiveness in gas separation is provided.

  10. Meniscus membranes for separations

    DOEpatents

    Dye, Robert C.; Jorgensen, Betty; Pesiri, David R.

    2004-01-27

    Gas separation membranes, especially meniscus-shaped membranes for gas separations are disclosed together with the use of such meniscus-shaped membranes for applications such as thermal gas valves, pre-concentration of a gas stream, and selective pre-screening of a gas stream. In addition, a rapid screening system for simultaneously screening polymer materials for effectiveness in gas separation is provided.

  11. Water vapor diffusion membranes

    NASA Technical Reports Server (NTRS)

    Holland, F. F., Jr.; Smith, J. K.

    1974-01-01

    The program is reported, which was designed to define the membrane technology of the vapor diffusion water recovery process and to test this technology using commercially available or experimental membranes. One membrane was selected, on the basis of the defined technology, and was subjected to a 30-day demonstration trial.

  12. Polyphosphazene semipermeable membranes

    DOEpatents

    Allen, Charles A.; McCaffrey, Robert R.; Cummings, Daniel G.; Grey, Alan E.; Jessup, Janine S.; McAtee, Richard E.

    1988-01-01

    A semipermeable, inorganic membrane is disclosed; the membrane is prepared from a phosphazene polymer and, by the selective substitution of the constituent groups bound to the phosphorous in the polymer structure, the selective passage of fluid from a feedstream can be controlled. Resistance to high temperatures and harsh chemical environments is observed in the use of the phosphazene polymers as semipermeable membranes.

  13. Graphene microsheets enter cells through spontaneous membrane penetration at edge asperities and corner sites

    PubMed Central

    Li, Yinfeng; Yuan, Hongyan; von dem Bussche, Annette; Creighton, Megan; Hurt, Robert H.; Kane, Agnes B.; Gao, Huajian

    2013-01-01

    Understanding and controlling the interaction of graphene-based materials with cell membranes is key to the development of graphene-enabled biomedical technologies and to the management of graphene health and safety issues. Very little is known about the fundamental behavior of cell membranes exposed to ultrathin 2D synthetic materials. Here we investigate the interactions of graphene and few-layer graphene (FLG) microsheets with three cell types and with model lipid bilayers by combining coarse-grained molecular dynamics (MD), all-atom MD, analytical modeling, confocal fluorescence imaging, and electron microscopic imaging. The imaging experiments show edge-first uptake and complete internalization for a range of FLG samples of 0.5- to 10-μm lateral dimension. In contrast, the simulations show large energy barriers relative to kBT for membrane penetration by model graphene or FLG microsheets of similar size. More detailed simulations resolve this paradox by showing that entry is initiated at corners or asperities that are abundant along the irregular edges of fabricated graphene materials. Local piercing by these sharp protrusions initiates membrane propagation along the extended graphene edge and thus avoids the high energy barrier calculated in simple idealized MD simulations. We propose that this mechanism allows cellular uptake of even large multilayer sheets of micrometer-scale lateral dimension, which is consistent with our multimodal bioimaging results for primary human keratinocytes, human lung epithelial cells, and murine macrophages. PMID:23840061

  14. Membrane tubule formation by banana-shaped proteins with or without transient network structure

    PubMed Central

    Noguchi, Hiroshi

    2016-01-01

    In living cells, membrane morphology is regulated by various proteins. Many membrane reshaping proteins contain a Bin/Amphiphysin/Rvs (BAR) domain, which consists of a banana-shaped rod. The BAR domain bends the biomembrane along the rod axis and the features of this anisotropic bending have recently been studied. Here, we report on the role of the BAR protein rods in inducing membrane tubulation, using large-scale coarse-grained simulations. We reveal that a small spontaneous side curvature perpendicular to the rod can drastically alter the tubulation dynamics at high protein density, whereas no significant difference is obtained at low density. A percolated network is intermediately formed depending on the side curvature. This network suppresses tubule protrusion, leading to the slow formation of fewer tubules. Thus, the side curvature, which is generated by protein–protein and membrane–protein interactions, plays a significant role in tubulation dynamics. We also find that positive surface tensions and the vesicle membrane curvature can stabilize this network structure by suppressing the tubulation. PMID:26863901

  15. Membrane tubule formation by banana-shaped proteins with or without transient network structure

    NASA Astrophysics Data System (ADS)

    Noguchi, Hiroshi

    2016-02-01

    In living cells, membrane morphology is regulated by various proteins. Many membrane reshaping proteins contain a Bin/Amphiphysin/Rvs (BAR) domain, which consists of a banana-shaped rod. The BAR domain bends the biomembrane along the rod axis and the features of this anisotropic bending have recently been studied. Here, we report on the role of the BAR protein rods in inducing membrane tubulation, using large-scale coarse-grained simulations. We reveal that a small spontaneous side curvature perpendicular to the rod can drastically alter the tubulation dynamics at high protein density, whereas no significant difference is obtained at low density. A percolated network is intermediately formed depending on the side curvature. This network suppresses tubule protrusion, leading to the slow formation of fewer tubules. Thus, the side curvature, which is generated by protein–protein and membrane–protein interactions, plays a significant role in tubulation dynamics. We also find that positive surface tensions and the vesicle membrane curvature can stabilize this network structure by suppressing the tubulation.

  16. Graphene microsheets enter cells through spontaneous membrane penetration at edge asperities and corner sites.

    PubMed

    Li, Yinfeng; Yuan, Hongyan; von dem Bussche, Annette; Creighton, Megan; Hurt, Robert H; Kane, Agnes B; Gao, Huajian

    2013-07-23

    Understanding and controlling the interaction of graphene-based materials with cell membranes is key to the development of graphene-enabled biomedical technologies and to the management of graphene health and safety issues. Very little is known about the fundamental behavior of cell membranes exposed to ultrathin 2D synthetic materials. Here we investigate the interactions of graphene and few-layer graphene (FLG) microsheets with three cell types and with model lipid bilayers by combining coarse-grained molecular dynamics (MD), all-atom MD, analytical modeling, confocal fluorescence imaging, and electron microscopic imaging. The imaging experiments show edge-first uptake and complete internalization for a range of FLG samples of 0.5- to 10-μm lateral dimension. In contrast, the simulations show large energy barriers relative to kBT for membrane penetration by model graphene or FLG microsheets of similar size. More detailed simulations resolve this paradox by showing that entry is initiated at corners or asperities that are abundant along the irregular edges of fabricated graphene materials. Local piercing by these sharp protrusions initiates membrane propagation along the extended graphene edge and thus avoids the high energy barrier calculated in simple idealized MD simulations. We propose that this mechanism allows cellular uptake of even large multilayer sheets of micrometer-scale lateral dimension, which is consistent with our multimodal bioimaging results for primary human keratinocytes, human lung epithelial cells, and murine macrophages.

  17. Tracking Membrane Protein Association in Model Membranes

    PubMed Central

    Reffay, Myriam; Gambin, Yann; Benabdelhak, Houssain; Phan, Gilles; Taulier, Nicolas; Ducruix, Arnaud; Hodges, Robert S.; Urbach, Wladimir

    2009-01-01

    Membrane proteins are essential in the exchange processes of cells. In spite of great breakthrough in soluble proteins studies, membrane proteins structures, functions and interactions are still a challenge because of the difficulties related to their hydrophobic properties. Most of the experiments are performed with detergent-solubilized membrane proteins. However widely used micellar systems are far from the biological two-dimensions membrane. The development of new biomimetic membrane systems is fundamental to tackle this issue. We present an original approach that combines the Fluorescence Recovery After fringe Pattern Photobleaching technique and the use of a versatile sponge phase that makes it possible to extract crucial informations about interactions between membrane proteins embedded in the bilayers of a sponge phase. The clear advantage lies in the ability to adjust at will the spacing between two adjacent bilayers. When the membranes are far apart, the only possible interactions occur laterally between proteins embedded within the same bilayer, whereas when membranes get closer to each other, interactions between proteins embedded in facing membranes may occur as well. After validating our approach on the streptavidin-biotinylated peptide complex, we study the interactions between two membrane proteins, MexA and OprM, from a Pseudomonas aeruginosa efflux pump. The mode of interaction, the size of the protein complex and its potential stoichiometry are determined. In particular, we demonstrate that: MexA is effectively embedded in the bilayer; MexA and OprM do not interact laterally but can form a complex if they are embedded in opposite bilayers; the population of bound proteins is at its maximum for bilayers separated by a distance of about 200 Å, which is the periplasmic thickness of Pseudomonas aeruginosa. We also show that the MexA-OprM association is enhanced when the position and orientation of the protein is restricted by the bilayers. We

  18. Mixed matrix membrane development.

    PubMed

    Kulprathipanja, Santi

    2003-03-01

    Two types of mixed matrix membranes were developed by UOP in the late 1980s. The first type includes adsorbent polymers, such as silicalite-cellulose acetate (CA), NaX-CA, and AgX-CA mixed matrix membranes. The silicalite-CA has a CO(2)/H(2) selectivity of 5.15 +/- 2.2. In contrast, the CA membrane has a CO(2)/H(2) selectivity of 0.77 +/- 0.06. The second type of mixed matrix membrane is PEG-silicone rubber. The PEG-silicone rubber mixed matrix membrane has high selectivity for polar gases, such as SO(2), NH(3), and H(2)S.

  19. Elastic membranes in confinement

    NASA Astrophysics Data System (ADS)

    Bostwick, Joshua; Miksis, Michael; Davis, Stephen

    2014-11-01

    An elastic membrane stretched between two walls takes a shape defined by its length and the volume of fluid it encloses. Many biological structures, such as cells, mitochondria and DNA, have finer internal structure in which a membrane (or elastic member) is geometrically ``confined'' by another object. We study the shape stability of elastic membranes in a ``confining'' box and introduce repulsive van der Waals forces to prevent the membrane from intersecting the wall. We aim to define the parameter space associated with mitochondria-like deformations. We compare the confined to `unconfined' solutions and show how the structure and stability of the membrane shapes changes with the system parameters.

  20. Separation membrane development

    SciTech Connect

    Lee, M.W.

    1998-08-01

    A ceramic membrane has been developed to separate hydrogen from other gases. The method used is a sol-gel process. A thin layer of dense ceramic material is coated on a coarse ceramic filter substrate. The pore size distribution in the thin layer is controlled by a densification of the coating materials by heat treatment. The membrane has been tested by permeation measurement of the hydrogen and other gases. Selectivity of the membrane has been achieved to separate hydrogen from carbon monoxide. The permeation rate of hydrogen through the ceramic membrane was about 20 times larger than Pd-Ag membrane.

  1. Membrane separation processes

    SciTech Connect

    Rautenbach, R.; Albrecht, R.

    1989-01-01

    The success of two membrane processes, reverse osmosis and ultrafiltration, has helped make membrane processes a central technique in solving separation problems for fluid systems. This book discusses the various applications and developments in membrane technology and shows how accurate membrane processes can be designed. Starting with the local transport phenomena, the behavior of individual elements such as tube or plate membrane and the behavior of the technical unit - the module - are discussed in detail. The book goes on to demonstrate the most effective ways of arranging modules for forming an optimal plant.

  2. Water Membrane Evaporator

    NASA Technical Reports Server (NTRS)

    Ungar, Eugene K.; Almlie, Jay C.

    2010-01-01

    A water membrane evaporator (WME) has been conceived and tested as an alternative to the contamination-sensitive and corrosion-prone evaporators currently used for dissipating heat from space vehicles. The WME consists mainly of the following components: An outer stainless-steel screen that provides structural support for the components mentioned next; Inside and in contact with the stainless-steel screen, a hydrophobic membrane that is permeable to water vapor; Inside and in contact with the hydrophobic membrane, a hydrophilic membrane that transports the liquid feedwater to the inner surface of the hydrophobic membrane; Inside and in contact with the hydrophilic membrane, an annular array of tubes through which flows the spacecraft coolant carrying the heat to be dissipated; and An inner exclusion tube that limits the volume of feedwater in the WME. In operation, a pressurized feedwater reservoir is connected to the volume between the exclusion tube and the coolant tubes. Feedwater fills the volume, saturates the hydrophilic membrane, and is retained by the hydrophobic membrane. The outside of the WME is exposed to space vacuum. Heat from the spacecraft coolant is conducted through the tube walls and the water-saturated hydrophilic membrane to the liquid/vapor interface at the hydrophobic membrane, causing water to evaporate to space. Makeup water flows into the hydrophilic membrane through gaps between the coolant tubes.

  3. Enhanced membrane gas separations

    SciTech Connect

    Prasad, R.

    1993-07-13

    An improved membrane gas separation process is described comprising: (a) passing a feed gas stream to the non-permeate side of a membrane system adapted for the passage of purge gas on the permeate side thereof, and for the passage of the feed gas stream in a counter current flow pattern relative to the flow of purge gas on the permeate side thereof, said membrane system being capable of selectively permeating a fast permeating component from said feed gas, at a feed gas pressure at or above atmospheric pressure; (b) passing purge gas to the permeate side of the membrane system in counter current flow to the flow of said feed gas stream in order to facilitate carrying away of said fast permeating component from the surface of the membrane and maintaining the driving force for removal of the fast permeating component through the membrane from the feed gas stream, said permeate side of the membrane being maintained at a subatmospheric pressure within the range of from about 0.1 to about 5 psia by vacuum pump means; (c) recovering a product gas stream from the non-permeate side of the membrane; and (d) discharging purge gas and the fast permeating component that has permeated the membrane from the permeate side of the membrane, whereby the vacuum conditions maintained on the permeate side of the membrane by said vacuum pump means enhance the efficiency of the gas separation operation, thereby reducing the overall energy requirements thereof.

  4. Catalytic membranes beckon

    SciTech Connect

    Caruana, C.M.

    1994-11-01

    Chemical engineers here and abroad are finding that the marriage of catalysts and membranes holds promise for faster and more specific reactions, although commercialization of this technology is several years away. Catalytic membrane reactors (CMRs) combine a heterogeneous catalyst and a permselective membrane. Reactions performed by CMRs provide higher yields--sometimes as much as 50% higher--because of better reaction selectivity--as opposed to separation selectivity. CMRs also can work at very high temperatures, using ceramic materials that would not be possible with organic membranes. Although the use of CMRs is not widespread presently, the development of new membranes--particularly porous ceramic and zeolite membranes--will increase the potential to improve yields of many catalytic processes. The paper discusses ongoing studies, metal and advanced materials for membranes, the need for continued research, hydrogen recovery from coal-derived gases, catalytic oxidation of sulfides, CMRs for water purification, and oxidative coupling of methane.

  5. Three dimensional microscopic surface profiles of membranes reconstructed from freeze etching electrol micrographs.

    PubMed

    Krbecek, R; Gebhardt, C; Gruler, H; Sackmann, E

    1979-06-13

    A method of three-dimensional reconstruction of the surface profile of artificial and natural membranes from freeze quenched electron micrographs is presented. The method is based on the analysis of the variation in thickness of platinum layers, deposited under an oblique angle. In essence, it is reminiscent of the method of Eratosthenes to measure the earth's radius. The thickness of etch-like protrusions of membranes could be determined to an accuracy of about 3 A. True distances on curved surfaces rather than projections of distances are obtained. The method has been applied to both model membranes and biological membranes. The essential results are: 1. Detailed information on the symmetry and the molecular structure of the crystalline phases of dimyristoyl phosphatidylcholine was obtained. The microscopic surface profile of the ripple structure observed between the pretransition and the main transition was analysed. In accordance with a previous model we found that the ripple structure is caused by the spontaneous curvature of the monolayers. The surface profiles of the ripple structure and of the low temperature biaxial phase could be clearly distinguished. 2. The sizes and shapes of lipid domains formed by both thermically and charge-induced lateral phase separation were determined. This showed that the visual inspection of electron micrographs may lead to a considerable underestimation of the domain size. Conclusions may be drawn concerning the different phases formed upon lateral phase separation. 3. As a biological example, yeast cell membranes were studied. The method allows one to distinguish between different membrane-bound proteins by measuring the width-to-height ratio of the particles. The deformation of the lipid layer in the environment of the proteins may be determined. This deformation contains information about lipid-mediated long-range interactions between membrane proteins. PMID:378255

  6. Three dimensional microscopic surface profiles of membranes reconstructed from freeze etching electrol micrographs.

    PubMed

    Krbecek, R; Gebhardt, C; Gruler, H; Sackmann, E

    1979-06-13

    A method of three-dimensional reconstruction of the surface profile of artificial and natural membranes from freeze quenched electron micrographs is presented. The method is based on the analysis of the variation in thickness of platinum layers, deposited under an oblique angle. In essence, it is reminiscent of the method of Eratosthenes to measure the earth's radius. The thickness of etch-like protrusions of membranes could be determined to an accuracy of about 3 A. True distances on curved surfaces rather than projections of distances are obtained. The method has been applied to both model membranes and biological membranes. The essential results are: 1. Detailed information on the symmetry and the molecular structure of the crystalline phases of dimyristoyl phosphatidylcholine was obtained. The microscopic surface profile of the ripple structure observed between the pretransition and the main transition was analysed. In accordance with a previous model we found that the ripple structure is caused by the spontaneous curvature of the monolayers. The surface profiles of the ripple structure and of the low temperature biaxial phase could be clearly distinguished. 2. The sizes and shapes of lipid domains formed by both thermically and charge-induced lateral phase separation were determined. This showed that the visual inspection of electron micrographs may lead to a considerable underestimation of the domain size. Conclusions may be drawn concerning the different phases formed upon lateral phase separation. 3. As a biological example, yeast cell membranes were studied. The method allows one to distinguish between different membrane-bound proteins by measuring the width-to-height ratio of the particles. The deformation of the lipid layer in the environment of the proteins may be determined. This deformation contains information about lipid-mediated long-range interactions between membrane proteins.

  7. Feedback Interactions of Polymerized Actin with the Cell Membrane: Waves, Pulses, and Oscillations

    NASA Astrophysics Data System (ADS)

    Carlsson, Anders

    Polymerized filaments of the protein actin have crucial functions in cell migration, and in bending the cell membrane to drive endocytosis or the formation of protrusions. The nucleation and polymerization of actin filaments are controlled by upstream agents in the cell membrane, including nucleation-promoting factors (NPFs) that activate the Arp2/3 complex to form new branches on pre-existing filaments. But polymerized actin (F-actin) also feeds back on the assembly of NPFs. We explore the effects of the resulting feedback loop of F-actin and NPFs on two phenomena: actin pulses that drive endocytosis in yeast, and actin waves traveling along the membrane of several cell types. In our model of endocytosis in yeast, the actin network is grown explicitly in three dimensions, exerts a negative feedback interaction on localized patch of NPFs in the membrane, and bends the membrane by exerting a distribution of forces. This model explains observed actin and NPF pulse dynamics, and the effects of several interventions including i) NPF mutations, ii) inhibition of actin polymerization, and iii) deletion of a protein that allows F-actin to bend the cell membrane. The model predicts that mutation of the active region of an NPF will enhance the accumulation of that NPF, and we confirm this prediction by quantitative fluorescence microscopy. For actin waves, we treat a similar model, with NPFs distributed over a larger region of the cell membrane. This model naturally generates actin waves, and predicts a transition from wave behavior to spatially localized oscillations when NPFs are confined to a small region. We also predict a transition from waves to static polarization as the negative-feedback coupling between F-actin and the NPFs is reduced. Supported by NIGMS Grant R01 GM107667.

  8. Membrane binding properties of IRSp53-missing in metastasis domain (IMD) protein.

    PubMed

    Futó, Kinga; Bódis, Emőke; Machesky, Laura M; Nyitrai, Miklós; Visegrády, Balázs

    2013-11-01

    The 53-kDa insulin receptor substrate protein (IRSp53) organizes the actin cytoskeleton in response to stimulation of small GTPases, promoting the formation of cell protrusions such as filopodia and lamellipodia. IMD is the N-terminal 250 amino acid domain (IRSp53/MIM Homology Domain) of IRSp53 (also called I-BAR), which can bind to negatively charged lipid molecules. Overexpression of IMD induces filopodia formation in cells and purified IMD assembles finger-like protrusions in reconstituted lipid membranes. IMD was shown by several groups to bundle actin filaments, but other groups showed that it also binds to membranes. IMD binds to negatively charged lipid molecules with preference to clusters of PI(4,5)P2. Here, we performed a range of different in vitro fluorescence experiments to determine the binding properties of the IMD to phospholipids. We used different constructs of large unilamellar vesicles (LUVETs), containing neutral or negatively charged phospholipids. We found that IMD has a stronger binding interaction with negatively charged PI(4,5)P2 or PS lipids than PS/PC or neutral PC lipids. The equilibrium dissociation constant for the IMD-lipid interaction falls into the 78-170μM range for all the lipids tested. The solvent accessibility of the fluorescence labels on the IMD during its binding to lipids is also reduced as the lipids become more negatively charged. Actin affects the IMD-lipid interaction, depending on its polymerization state. Monomeric actin partially disrupts the binding, while filamentous actin can further stabilize the IMD-lipid interaction. PMID:23872532

  9. Membrane with supported internal passages

    NASA Technical Reports Server (NTRS)

    Gonzalez-Martin, Anuncia (Inventor); Salinas, Carlos E. (Inventor); Cisar, Alan J. (Inventor); Hitchens, G. Duncan (Inventor); Murphy, Oliver J. (Inventor)

    2000-01-01

    The invention provides an improved proton exchange membrane for use in electrochemical cells having internal passages parallel to the membrane surface comprising permanent tubes preferably placed at the ends of the fluid passages. The invention also provides an apparatus and process for making the membrane, membrane and electrode assemblies fabricated using the membrane, and the application of the membrane and electrode assemblies to a variety of devices, both electrochemical and otherwise. The passages in the membrane extend from one edge of the membrane to another and allow fluid flow through the membrane and give access directly to the membrane.

  10. Functions and Regulation of Circular Dorsal Ruffles

    PubMed Central

    Hoon, Jing-Ling; Wong, Wai-Keung

    2012-01-01

    Cells construct a number of plasma membrane structures to meet a range of physiological demands. Driven by juxtamembrane actin machinery, these actin-based membrane protrusions are essential for the operation and maintenance of cellular life. They are required for diverse cellular functions, such as directed cell motility, cell spreading, adhesion, and substrate/matrix degradation. Circular dorsal ruffles (CDRs) are one class of such structures characterized as F-actin-rich membrane projections on the apical cell surface. CDRs commence their formation minutes after stimulation as flat, open, and immature ruffles and progressively develop into fully enclosed circular ruffles. These “rings” then mature and contract centrifugally before subsiding. Serving a critical function in receptor internalization and cell migration, CDRs are thus highly dynamic but transient formations. Here, we review the current state of knowledge concerning the regulation of circular dorsal ruffles. We focus specifically on the biochemical pathways leading to CDR formation in order to better define the roles and functions of these enigmatic structures. PMID:22927640

  11. Single-molecule tracking of small GTPase Rac1 uncovers spatial regulation of membrane translocation and mechanism for polarized signaling

    PubMed Central

    Das, Sulagna; Yin, Taofei; Yang, Qingfen; Zhang, Jingqiao; Wu, Yi I.; Yu, Ji

    2015-01-01

    Polarized Rac1 signaling is a hallmark of many cellular functions, including cell adhesion, motility, and cell division. The two steps of Rac1 activation are its translocation to the plasma membrane and the exchange of nucleotide from GDP to GTP. It is, however, unclear whether these two processes are regulated independent of each other and what their respective roles are in polarization of Rac1 signaling. We designed a single-particle tracking (SPT) method to quantitatively analyze the kinetics of Rac1 membrane translocation in living cells. We found that the rate of Rac1 translocation was significantly elevated in protrusions during cell spreading on collagen. Furthermore, combining FRET sensor imaging with SPT measurements in the same cell, the recruitment of Rac1 was found to be polarized to an extent similar to that of the nucleotide exchange process. Statistical analysis of single-molecule trajectories and optogenetic manipulation of membrane lipids revealed that Rac1 membrane translocation precedes nucleotide exchange, and is governed primarily by interactions with phospholipids, particularly PI(3,4,5)P3, instead of protein factors. Overall, the study highlights the significance of membrane translocation in spatial Rac1 signaling, which is in addition to the traditional view focusing primarily on GEF distribution and exchange reaction. PMID:25561548

  12. Substituted polyacetylene separation membrane

    DOEpatents

    Pinnau, I.; Morisato, Atsushi

    1998-01-13

    A separation membrane is described which is useful for gas separation, particularly separation of C{sub 2+} hydrocarbons from natural gas. The invention encompasses the membrane itself, methods of making it and processes for using it. The membrane comprises a polymer having repeating units of a hydrocarbon-based, disubstituted polyacetylene, having the general formula shown in the accompanying diagram, wherein R{sub 1} is chosen from the group consisting of C{sub 1}-C{sub 4} alkyl and phenyl, and wherein R{sub 2} is chosen from the group consisting of hydrogen and phenyl. In the most preferred embodiment, the membrane comprises poly(4-methyl-2-pentyne) [PMP]. The membrane exhibits good chemical resistance and has super-glassy properties with regard to separating certain large, condensable permeant species from smaller, less-condensable permeant species. The membranes may also be useful in other fluid separations. 4 figs.

  13. Substituted polyacetylene separation membrane

    DOEpatents

    Pinnau, Ingo; Morisato, Atsushi

    1998-01-13

    A separation membrane useful for gas separation, particularly separation of C.sub.2+ hydrocarbons from natural gas. The invention encompasses the membrane itself, methods of making it and processes for using it. The membrane comprises a polymer having repeating units of a hydrocarbon-based, disubstituted polyacetylene, having the general formula: ##STR1## wherein R.sub.1 is chosen from the group consisting of C.sub.1 -C.sub.4 alkyl and phenyl, and wherein R.sub.2 is chosen from the group consisting of hydrogen and phenyl. In the most preferred embodiment, the membrane comprises poly(4-methyl-2-pentyne) ›PMP!. The membrane exhibits good chemical resistance and has super-glassy properties with regard to separating certain large, condensable permeant species from smaller, less-condensable permeant species. The membranes may also be useful in other fluid separations.

  14. Anion permselective membrane

    NASA Technical Reports Server (NTRS)

    Alexander, S.; Hodgdon, R. B.

    1977-01-01

    The objective of NAS 3-20108 was the development and evaluation of improved anion selective membranes useful as efficient separators in a redox power storage cell system being constructed. The program was divided into three parts, (a) optimization of the selected candidate membrane systems, (b) investigation of alternative membrane/polymer systems, and (c) characterization of candidate membranes. The major synthesis effort was aimed at improving and optimizing as far as possible each candidate system with respect to three critical membrane properties essential for good redox cell performance. Substantial improvements were made in 5 candidate membrane systems. The critical synthesis variables of cross-link density, monomer ratio, and solvent composition were examined over a wide range. In addition, eight alternative polymer systems were investigated, two of which attained candidate status. Three other alternatives showed potential but required further research and development. Each candidate system was optimized for selectivity.

  15. Poxvirus Membrane Biogenesis

    PubMed Central

    2015-01-01

    Poxviruses differ from most DNA viruses by replicating entirely within the cytoplasm. The first discernible viral structures are crescents and spherical immature virions containing a single lipoprotein membrane bilayer with an external honeycomb lattice. Because this viral membrane displays no obvious continuity with a cellular organelle, a de novo origin was suggested. Nevertheless, transient connections between viral and cellular membranes could be difficult to resolve. Despite the absence of direct evidence, the intermediate compartment (ERGIC) between the endoplasmic reticulum (ER) and Golgi apparatus and the ER itself were considered possible sources of crescent membranes. A break-through in understanding poxvirus membrane biogenesis has come from recent studies of the abortive replication of several vaccinia virus null mutants. Novel images showing continuity between viral crescents and the ER and the accumulation of immature virions in the expanded ER lumen provide the first direct evidence for a cellular origin of this poxvirus membrane. PMID:25728299

  16. Supported liquid membrane system

    SciTech Connect

    Takigawa, D.Y.; Bush, H. Jr.

    1990-12-31

    A cell apparatus for a supported liquid membrane including opposing faceplates, each having a spirally configured groove, an inlet groove at a first end of the spirally configured groove, and an outlet groove at the other end of the spirally configured groove, within the opposing faces of the faceplates, a microporous membrane situated between the grooved faces of the faceplates, said microporous membrane containing an extractant mixture selective for a predetermined chemical species within the pores of said membrane, means for aligning the grooves of the faceplates in an directly opposing configuration with the porous membrane being situated therebetween, such that the aligned grooves form a pair of directly opposing channels, separate feed solution and stripping solution compartments connected to respective channels between the faceplates and the membrane, separate pumping means for passing feed solution and stripping solution through the channels is provided.

  17. Anion permselective membrane

    NASA Technical Reports Server (NTRS)

    Hodgdon, R. B.; Waite, W. A.; Alexander, S. S.

    1984-01-01

    Two polymer ion exchange membranes were synthesized to fulfill the needs of both electrical resistivity and anolyte/catholyte separation for utility load leveling utilizing the DOE/NASA mixed electrolyte REDOX battery. Both membranes were shown to meet mixed electrolyte utility load leveling criteria. Several modifications of an anion exchange membrane failed to meet utility load leveling REDOX battery criteria using the unmixed electrolyte REDOX cell.

  18. Siloxane-grafted membranes

    DOEpatents

    Friesen, Dwayne T.; Obligin, Alan S.

    1989-01-01

    Composite cellulosic semipermeable membranes are disclosed which are the covalently bonded reaction product of an asymmetric cellulosic semipermeable membrane and a polysiloxane containing reactive functional groups. The two reactants chemically bond by ether, ester, amide or acrylate linkages to form a siloxane-grafted cellulosic membrane having superior selectivity and flux stability. Selectivity may be enhanced by wetting the surface with a swelling agent such as water.

  19. Polyarylether composition and membrane

    DOEpatents

    Hung, Joyce; Brunelle, Daniel Joseph; Harmon, Marianne Elisabeth; Moore, David Roger; Stone, Joshua James; Zhou, Hongyi; Suriano, Joseph Anthony

    2010-11-09

    A composition including a polyarylether copolymer is provided. The copolymer includes a polyarylether backbone; and a sulfonated oligomeric group bonded to the polyarylether suitable for use as a cation conducting membrane. Method of bonding a sulfonated oligomeric group to the polyarylether backbone to form a polyarylether copolymer. The membrane may be formed from the polyarylether copolymer composition. The chain length of the sulfonated oligomeric group may be controlled to affect or control the ion conductivity of the membrane.

  20. Anion permselective membrane

    NASA Astrophysics Data System (ADS)

    Hodgdon, R. B.; Waite, W. A.; Alexander, S. S.

    1984-07-01

    Two polymer ion exchange membranes were synthesized to fulfill the needs of both electrical resistivity and anolyte/catholyte separation for utility load leveling utilizing the DOE/NASA mixed electrolyte REDOX battery. Both membranes were shown to meet mixed electrolyte utility load leveling criteria. Several modifications of an anion exchange membrane failed to meet utility load leveling REDOX battery criteria using the unmixed electrolyte REDOX cell.

  1. Siloxane-grafted membranes

    DOEpatents

    Friesen, D.T.; Obligin, A.S.

    1989-10-31

    Composite cellulosic semipermeable membranes are disclosed which are the covalently bonded reaction product of an asymmetric cellulosic semipermeable membrane and a polysiloxane containing reactive functional group. The two reactants chemically bond by ether, ester, amide or acrylate linkages to form a siloxane-grafted cellulosic membrane having superior selectivity and flux stability. Selectivity may be enhanced by wetting the surface with a swelling agent such as water.

  2. Proteins causing membrane fouling in membrane bioreactors.

    PubMed

    Miyoshi, Taro; Nagai, Yuhei; Aizawa, Tomoyasu; Kimura, Katsuki; Watanabe, Yoshimasa

    2015-01-01

    In this study, the details of proteins causing membrane fouling in membrane bioreactors (MBRs) treating real municipal wastewater were investigated. Two separate pilot-scale MBRs were continuously operated under significantly different operating conditions; one MBR was a submerged type whereas the other was a side-stream type. The submerged and side-stream MBRs were operated for 20 and 10 days, respectively. At the end of continuous operation, the foulants were extracted from the fouled membranes. The proteins contained in the extracted foulants were enriched by using the combination of crude concentration with an ultrafiltration membrane and trichloroacetic acid precipitation, and then separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). The N-terminal amino acid sequencing analysis of the proteins which formed intensive spots on the 2D-PAGE gels allowed us to partially identify one protein (OmpA family protein originated from genus Brevundimonas or Riemerella anatipestifer) from the foulant obtained from the submerged MBR, and two proteins (OprD and OprF originated from genus Pseudomonas) from that obtained from the side-stream MBR. Despite the significant difference in operating conditions of the two MBRs, all proteins identified in this study belong to β-barrel protein. These findings strongly suggest the importance of β-barrel proteins in developing membrane fouling in MBRs.

  3. Proteins causing membrane fouling in membrane bioreactors.

    PubMed

    Miyoshi, Taro; Nagai, Yuhei; Aizawa, Tomoyasu; Kimura, Katsuki; Watanabe, Yoshimasa

    2015-01-01

    In this study, the details of proteins causing membrane fouling in membrane bioreactors (MBRs) treating real municipal wastewater were investigated. Two separate pilot-scale MBRs were continuously operated under significantly different operating conditions; one MBR was a submerged type whereas the other was a side-stream type. The submerged and side-stream MBRs were operated for 20 and 10 days, respectively. At the end of continuous operation, the foulants were extracted from the fouled membranes. The proteins contained in the extracted foulants were enriched by using the combination of crude concentration with an ultrafiltration membrane and trichloroacetic acid precipitation, and then separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE). The N-terminal amino acid sequencing analysis of the proteins which formed intensive spots on the 2D-PAGE gels allowed us to partially identify one protein (OmpA family protein originated from genus Brevundimonas or Riemerella anatipestifer) from the foulant obtained from the submerged MBR, and two proteins (OprD and OprF originated from genus Pseudomonas) from that obtained from the side-stream MBR. Despite the significant difference in operating conditions of the two MBRs, all proteins identified in this study belong to β-barrel protein. These findings strongly suggest the importance of β-barrel proteins in developing membrane fouling in MBRs. PMID:26360742

  4. Gas separation membranes

    DOEpatents

    Schell, William J.

    1979-01-01

    A dry, fabric supported, polymeric gas separation membrane, such as cellulose acetate, is prepared by casting a solution of the polymer onto a shrinkable fabric preferably formed of synthetic polymers such as polyester or polyamide filaments before washing, stretching or calendering (so called griege goods). The supported membrane is then subjected to gelling, annealing, and drying by solvent exchange. During the processing steps, both the fabric support and the membrane shrink a preselected, controlled amount which prevents curling, wrinkling or cracking of the membrane in flat form or when spirally wound into a gas separation element.

  5. Biomolecular membrane protein crystallization

    NASA Astrophysics Data System (ADS)

    Reddy Bolla, Jani; Su, Chih-Chia; Yu, Edward W.

    2012-07-01

    Integral membrane proteins comprise approximately 30% of the sequenced genomes, and there is an immediate need for their high-resolution structural information. Currently, the most reliable approach to obtain these structures is X-ray crystallography. However, obtaining crystals of membrane proteins that diffract to high resolution appears to be quite challenging, and remains a major obstacle in structural determination. This brief review summarizes a variety of methodologies for use in crystallizing these membrane proteins. Hopefully, by introducing the available methods, techniques, and providing a general understanding of membrane proteins, a rational decision can be made about now to crystallize these complex materials.

  6. Anion exchange membrane

    DOEpatents

    Verkade, John G; Wadhwa, Kuldeep; Kong, Xueqian; Schmidt-Rohr, Klaus

    2013-05-07

    An anion exchange membrane and fuel cell incorporating the anion exchange membrane are detailed in which proazaphosphatrane and azaphosphatrane cations are covalently bonded to a sulfonated fluoropolymer support along with anionic counterions. A positive charge is dispersed in the aforementioned cations which are buried in the support to reduce the cation-anion interactions and increase the mobility of hydroxide ions, for example, across the membrane. The anion exchange membrane has the ability to operate at high temperatures and in highly alkaline environments with high conductivity and low resistance.

  7. Membrane Tension Control

    NASA Technical Reports Server (NTRS)

    Su, Ji (Inventor); Harrison, Joycelyn S. (Inventor)

    2005-01-01

    An electrostrictive polymer actuator comprises an electrostrictive polymer with a tailorable Poisson's ratio. The electrostrictive polymer is electroded on its upper and lower surfaces and bonded to an upper material layer. The assembly is rolled tightly and capped at its ends. In a membrane structure having a membrane, a supporting frame and a plurality of threads connecting the membrane to the frame, an actuator can be integrated into one or more of the plurality of threads. The electrostrictive polymer actuator displaces along its longitudinal axis, thereby affecting movement of the membrane surface.

  8. A Non-sulfated Chondroitin Stabilizes Membrane Tubulation in Cnidarian Organelles*

    PubMed Central

    Adamczyk, Patrizia; Zenkert, Claudia; Balasubramanian, Prakash G.; Yamada, Shuhei; Murakoshi, Saori; Sugahara, Kazuyuki; Hwang, Jung Shan; Gojobori, Takashi; Holstein, Thomas W.; Özbek, Suat

    2010-01-01

    Membrane tubulation is generally associated with rearrangements of the cytoskeleton and other cytoplasmic factors. Little is known about the contribution of extracellular matrix components to this process. Here, we demonstrate an essential role of proteoglycans in the tubulation of the cnidarian nematocyst vesicle. The morphogenesis of this extrusive organelle takes place inside a giant post-Golgi vesicle, which topologically represents extracellular space. This process includes the formation of a complex collagenous capsule structure that elongates into a long tubule, which invaginates after its completion. We show that a non-sulfated chondroitin appears as a scaffold in early morphogenesis of all nematocyst types in Hydra and Nematostella. It accompanies the tubulation of the vesicle membrane forming a provisional tubule structure, which after invagination matures by collagen incorporation. Inhibition of chondroitin synthesis by β-xylosides arrests nematocyst morphogenesis at different stages of tubule outgrowth resulting in retention of tubule material and a depletion of mature capsules in the tentacles of hydra. Our data suggest a conserved role of proteoglycans in the stabilization of a membrane protrusion as an essential step in organelle morphogenesis. PMID:20538610

  9. Membrane Degradation Accelerated Stress Test

    SciTech Connect

    Mukundan, Rangachary; Borup, Rodney L.

    2015-01-21

    These are a set of slides that deal with membrane degradation accelerated stress test. Specifically, the following topics are covered: membrane degradation FCTT drive cycle; membrane ASTs; current membrane ASTs damage mechanisms; proposed membrane AST, RH cycling in H2/Air; current proposed AST; 2min/2min AST damage mechanism; 30sec/45sec RH cycling at OCV.

  10. Permeable membrane experiment

    NASA Technical Reports Server (NTRS)

    Slavin, Thomas J.; Cao, Tuan Q.; Kliss, Mark H.

    1993-01-01

    The purpose of the Permeable Membrane Experiment is to gather flight data on three areas of membrane performance that are influenced by the presence of gravity. These areas are: (1) Liquid/gas phase separation, (2) gas bubble interference with diffusion through porous membranes and (3) wetting characteristics of hydrophilic membrane surfaces. These data are important in understaning the behavior of membrane/liquid/gas interfaces where surface tension forces predominate. The data will be compared with 1-g data already obtained and with predicted micrograviity behavior. The data will be used to develop designs for phase separation and plant nutrient delivery systems and will be available to the life support community for use in developing technologies which employ membranes. A conceptual design has been developed to conduct three membrane experiments, in sequence, aboard a single Complex Autonomous Payload (CAP) carrier to be carried in the Shuttle Orbiter payload bay. One experiment is conducted for each of the three membrane performance areas under study. These experiments are discussed in this paper.

  11. Membrane Transport Phenomena (MTP)

    NASA Technical Reports Server (NTRS)

    Mason, Larry W.

    1996-01-01

    The development of the seal between the membrane and the Fluid Optical Cells (FOC) has been a high priority activity. This seal occurs at an interface in the instrument where three key functions must be realized: (1) physical membrane support, (2) fluid sealing, and (3) unobscured optical transmission.

  12. Microporous alumina ceramic membranes

    DOEpatents

    Anderson, Marc A.; Sheng, Guangyao

    1993-01-01

    Several methods are disclosed for the preparation microporous alumina ceramic membranes. For the first time, porous alumina membranes are made which have mean pore sizes less than 100 Angstroms and substantially no pores larger than that size. The methods are based on improved sol-gel techniques.

  13. Membrane in cancer cells

    SciTech Connect

    Galeotti, T.; Cittadini, A.; Neri, G.; Scarpa, A.

    1988-01-01

    This book contains papers presented at a conference on membranes in cancer cells. Topics covered include Oncogenies, hormones, and free-radical processes in malignant transformation in vitro and Superoxide onion may trigger DNA strand breaks in human granulorytes by acting as a membrane target.

  14. Membranebioreactor with external membranes.

    PubMed

    Jonkers, C; Carrette, R; De Lathouwer, J; Thoeye, C

    2001-01-01

    The recent development of less expensive and more performant ultrafiltration membranes has generated a new concept in biological treatment known as membrane bioreactors (MBR). In the field of wastewater treatment, the MBR process is used as a modification of the conventional activated sludge (CAS) process, where the clarifier is replaced by a membrane module for the separation of the solid and the liquid phase. Membrane technology can, amongst others, be applied for the fast retrofit of relatively high loaded activated sludge systems to nutrient removal. In the last year Aquafin has built up extensive experience with submerged membrane systems. The research presented here was aimed at evaluating a particular external tubular membrane unit for MBR. More specifically it was evaluated whether higher stable flux levels could compensate the higher energy consumption, typical for external membrane systems. Both (1) treatment performance and (2) possible operational drawbacks were tested on a 5 m3/h activated sludge pilot-scale. The membrane bioreactor was tested in parallel with a conventional activated sludge system.

  15. Membrane module assembly

    DOEpatents

    Kaschemekat, Jurgen

    1994-01-01

    A membrane module assembly adapted to provide a flow path for the incoming feed stream that forces it into prolonged heat-exchanging contact with a heating or cooling mechanism. Membrane separation processes employing the module assembly are also disclosed. The assembly is particularly useful for gas separation or pervaporation.

  16. Anion permselective membrane

    NASA Technical Reports Server (NTRS)

    Hodgdon, R. B.; Waite, W. A.

    1982-01-01

    The synthesis and fabrication of polymeric anion permselective membranes for redox systems are discussed. Variations of the prime candidate anion membrane formulation to achieve better resistance and/or lower permeability were explored. Processing parameters were evaluated to lower cost and fabricate larger sizes. The processing techniques to produce more membranes per batch were successfully integrated with the fabrication of larger membranes. Membranes of about 107 cm x 51 cm were made in excellent yield. Several measurements were made on the larger sample membranes. Among the data developed were water transport and transference numbers for these prime candidate membranes at 20 C. Other work done on this system included characterization of a number of specimens of candidate membranes which had been returned after service lives of up to sixteen months. Work with new polymer constituents, with new N.P.'s, catalysts and backing fabrics is discussed. Some work was also done to evaluate other proportions of the ingredients of the prime candidate system. The adoption of a flow selectivity test at elevated temperature was explored.

  17. Polymide gas separation membranes

    DOEpatents

    Ding, Yong; Bikson, Benjamin; Nelson, Joyce Katz

    2004-09-14

    Soluble polyamic acid salt (PAAS) precursors comprised of tertiary and quaternary amines, ammonium cations, sulfonium cations, or phosphonium cations, are prepared and fabricated into membranes that are subsequently imidized and converted into rigid-rod polyimide articles, such as membranes with desirable gas separation properties. A method of enhancing solubility of PAAS polymers in alcohols is also disclosed.

  18. Membrane module assembly

    DOEpatents

    Kaschemekat, J.

    1994-03-15

    A membrane module assembly is described which is adapted to provide a flow path for the incoming feed stream that forces it into prolonged heat-exchanging contact with a heating or cooling mechanism. Membrane separation processes employing the module assembly are also disclosed. The assembly is particularly useful for gas separation or pervaporation. 2 figures.

  19. Microporous alumina ceramic membranes

    DOEpatents

    Anderson, M.A.; Guangyao Sheng.

    1993-05-04

    Several methods are disclosed for the preparation microporous alumina ceramic membranes. For the first time, porous alumina membranes are made which have mean pore sizes less than 100 Angstroms and substantially no pores larger than that size. The methods are based on improved sol-gel techniques.

  20. Zeolite membranes from kaolin

    SciTech Connect

    Karle, B.G.; Brinker, C.J. |; Phillips, M.L.F.

    1996-07-01

    Zeolite films are sought as components of molecular sieve membranes. Different routes used to prepare zeolite composite membranes include growing zeolite layers from gels on porous supports, depositing oriented zeolites on supports, and dispersing zeolites in polymeric membranes. In most cases, it is very difficult to control and avoid the formation of cracks and/or pinholes. The approach to membrane synthesis is based on hydrothermally converting films of layered aluminosilicates into zeolite films. The authors have demonstrated this concept by preparing zeolite A membranes on alumina supports from kaolin films. The authors have optimized the process parameters not only for desired bulk properties, but also for preparing thin (ca. 5 {micro}m), continuous zeolite A films. Scanning electron microscopy shows highly intergrown zeolite A crystals over most of the surface area of the membrane, but gas permeation experiments indicate existence of mesoporous defects and/or intercrystalline gaps. It has been demonstrated that the thickness of the final zeolite A membrane can be controlled by limiting the amount of precursor kaolin present in the membrane.

  1. Elastic membranes in confinement.

    PubMed

    Bostwick, J B; Miksis, M J; Davis, S H

    2016-07-01

    An elastic membrane stretched between two walls takes a shape defined by its length and the volume of fluid it encloses. Many biological structures, such as cells, mitochondria and coiled DNA, have fine internal structure in which a membrane (or elastic member) is geometrically 'confined' by another object. Here, the two-dimensional shape of an elastic membrane in a 'confining' box is studied by introducing a repulsive confinement pressure that prevents the membrane from intersecting the wall. The stage is set by contrasting confined and unconfined solutions. Continuation methods are then used to compute response diagrams, from which we identify the particular membrane mechanics that generate mitochondria-like shapes. Large confinement pressures yield complex response diagrams with secondary bifurcations and multiple turning points where modal identities may change. Regions in parameter space where such behaviour occurs are then mapped. PMID:27440257

  2. Anton permselective membrane

    NASA Technical Reports Server (NTRS)

    Alexander, S. S.; Hodgdon, R. B.; Waite, W. A.

    1979-01-01

    Experimental composite membranes were synthesized on a lab scale consisting of a thin layer of anion permselective resin supported by and bonded to a porous physically strong and conductive substrate film. These showed good selectivity and also substantially lower electrical resistivities than the homogenous candidate membranes optimized in the previous contract. A wide range of resin porosities were examined for three candidate membrane systems, CDIL, CP4L, and A3L to identify the formulation giving the best overall redox cell performance. Candidate anion membranes showed large increases in resistivity after a short time of immersion in concentrated FeCl/HCl solution. Largely on the basis of resistance stability the CDIL formulation was selected as prime candidate and about thirty-five membranes (one foot square) were produced for experimental static and dynamic evaluation.

  3. Viral Membrane Scission

    PubMed Central

    Rossman, Jeremy S.; Lamb, Robert A.

    2014-01-01

    Virus budding is a complex, multistep process in which viral proteins make specific alterations in membrane curvature. Many different viral proteins can deform the membrane and form a budding virion, but very few can mediate membrane scission to complete the budding process. As a result, enveloped viruses have developed numerous ways of facilitating membrane scission, including hijacking host cellular scission machinery and expressing their own scission proteins. These proteins mediate scission in very different ways, though the biophysical mechanics underlying their actions may be similar. In this review, we explore the mechanisms of membrane scission and the ways in which enveloped viruses use these systems to mediate the release of budding virions. PMID:24099087

  4. Ion-conducting membranes

    DOEpatents

    Masel, Richard L.; Chen, Qingmei; Liu, Zengcai; Kutz, Robert

    2016-06-21

    An ion conducting polymeric composition mixture comprises a copolymer of styrene and vinylbenzyl-R.sub.s. R.sub.s is selected from the group consisting of imidazoliums and pyridiniums. The composition contains 10%-90% by weight of vinylbenzyl-R.sub.s. The composition can further comprise a polyolefin comprising substituted polyolefins, a polymer comprising cyclic amine groups, a polymer comprising at least one of a phenylene group and a phenyl group, a polyamide, and/or the reaction product of a constituent having two carbon-carbon double bonds. The composition can be in the form of a membrane. In a preferred embodiment, the membrane is a Helper Membrane that increases the faradaic efficiency of an electrochemical cell into which the membrane is incorporated, and also allows product formation at lower voltages than in cells without the Helper Membrane.

  5. Membrane projection lithography

    DOEpatents

    Burckel, David Bruce; Davids, Paul S; Resnick, Paul J; Draper, Bruce L

    2015-03-17

    The various technologies presented herein relate to a three dimensional manufacturing technique for application with semiconductor technologies. A membrane layer can be formed over a cavity. An opening can be formed in the membrane such that the membrane can act as a mask layer to the underlying wall surfaces and bottom surface of the cavity. A beam to facilitate an operation comprising any of implantation, etching or deposition can be directed through the opening onto the underlying surface, with the opening acting as a mask to control the area of the underlying surfaces on which any of implantation occurs, material is removed, and/or material is deposited. The membrane can be removed, a new membrane placed over the cavity and a new opening formed to facilitate another implantation, etching, or deposition operation. By changing the direction of the beam different wall/bottom surfaces can be utilized to form a plurality of structures.

  6. Alterations of red cell membrane properties in neuroacanthocytosis.

    PubMed

    Siegl, Claudia; Hamminger, Patricia; Jank, Herbert; Ahting, Uwe; Bader, Benedikt; Danek, Adrian; Gregory, Allison; Hartig, Monika; Hayflick, Susan; Hermann, Andreas; Prokisch, Holger; Sammler, Esther M; Yapici, Zuhal; Prohaska, Rainer; Salzer, Ulrich

    2013-01-01

    Neuroacanthocytosis (NA) refers to a group of heterogenous, rare genetic disorders, namely chorea acanthocytosis (ChAc), McLeod syndrome (MLS), Huntington's disease-like 2 (HDL2) and pantothenate kinase associated neurodegeneration (PKAN), that mainly affect the basal ganglia and are associated with similar neurological symptoms. PKAN is also assigned to a group of rare neurodegenerative diseases, known as NBIA (neurodegeneration with brain iron accumulation), associated with iron accumulation in the basal ganglia and progressive movement disorder. Acanthocytosis, the occurrence of misshaped erythrocytes with thorny protrusions, is frequently observed in ChAc and MLS patients but less prevalent in PKAN (about 10%) and HDL2 patients. The pathological factors that lead to the formation of the acanthocytic red blood cell shape are currently unknown. The aim of this study was to determine whether NA/NBIA acanthocytes differ in their functionality from normal erythrocytes. Several flow-cytometry-based assays were applied to test the physiological responses of the plasma membrane, namely drug-induced endocytosis, phosphatidylserine exposure and calcium uptake upon treatment with lysophosphatidic acid. ChAc red cell samples clearly showed a reduced response in drug-induced endovesiculation, lysophosphatidic acid-induced phosphatidylserine exposure, and calcium uptake. Impaired responses were also observed in acanthocyte-positive NBIA (PKAN) red cells but not in patient cells without shape abnormalities. These data suggest an "acanthocytic state" of the red cell where alterations in functional and interdependent membrane properties arise together with an acanthocytic cell shape. Further elucidation of the aberrant molecular mechanisms that cause this acanthocytic state may possibly help to evaluate the pathological pathways leading to neurodegeneration. PMID:24098554

  7. Rotating for elongation: Fat2 whips for the race.

    PubMed

    Stürner, Tomke; Tavosanis, Gaia

    2016-02-29

    Dynamic rearrangements of the actin cytoskeleton are crucial for cell shape and migration. In this issue, Squarr et al. (2016. J. Cell Biol. http://dx.doi.org/10.1083/jcb.201508081) show that the cadherin superfamily protein Fat2 regulates actin-rich protrusions driving collective cell migration during Drosophila melanogaster egg morphogenesis through its interaction with the WAVE regulatory complex. PMID:26903537

  8. Ethanol and membrane lipids.

    PubMed

    Sun, G Y; Sun, A Y

    1985-01-01

    Although ethanol is known to exert its primary mode of action on the central nervous system, the exact molecular interaction underlying the behavioral and physiological manifestations of alcohol intoxication has not been elucidated. Chronic ethanol administration results in changes in organ functions. These changes are reflective of the adaptive mechanisms in response to the acute effects of ethanol. Biophysical studies have shown that ethanol in vitro disorders the membrane and perturbs the fine structural arrangement of the membrane lipids. In the chronic state, these membranes develop resistance to the disordering effects. Tolerance development is also accompanied by biochemical changes. Although ethanol-induced changes in membrane lipids have been implicated in both biophysical and biochemical studies, measurements of membrane lipids, such as cholesterol content, fatty acid unsaturation, phospholipid distribution, and ganglioside profiles, have not produced conclusive evidence that any of these parameters are directly involved in the action of ethanol. On the other hand, there is increasing evidence indicating that although ethanol in vitro produces a membrane-fluidizing effect, the chronic response to this effect is not to change the membrane bulk lipid composition. Instead, changes in membrane lipids may pertain to small metabolically active pools located in certain subcellular fractions. Most likely, these lipids are involved in important membrane functions. For example, the increase in PS in brain plasma membranes may provide an explanation for the adaptive increase in synaptic membrane ion transport activity, especially (Na,K)-ATPase. There is also evidence that the lipid pool involved in the deacylation-reacylation mechanism (i.e., PI and PC with 20:4 groups) is altered after ethanol administration. An increase in metabolic turnover of these phospholipid pools may have important implications for the membrane functional changes. Obviously, there are other

  9. Microfiltration membranes: characteristics and manufacturing.

    PubMed

    Reif, Oscar W

    2006-01-01

    Membrane filtration is used within a multitude of processes ranging from dialysis to desalination processes to sterilizing filtration in the pharmaceutical industry. Membranes, nevertheless, have to have special characteristics and properties to serve such specific applications. Microfiltration membranes are utilized in a large range of membrane polymers and structures, which all have individual production process steps to achieve consistently the same membrane parameters. This chapter discusses membrane polymers and production processes in detail.

  10. Catalytic nanoporous membranes

    DOEpatents

    Pellin, Michael J; Hryn, John N; Elam, Jeffrey W

    2013-08-27

    A nanoporous catalytic membrane which displays several unique features Including pores which can go through the entire thickness of the membrane. The membrane has a higher catalytic and product selectivity than conventional catalysts. Anodic aluminum oxide (AAO) membranes serve as the catalyst substrate. This substrate is then subjected to Atomic Layer Deposition (ALD), which allows the controlled narrowing of the pores from 40 nm to 10 nm in the substrate by deposition of a preparatory material. Subsequent deposition of a catalytic layer on the inner surfaces of the pores reduces pore sizes to less than 10 nm and allows for a higher degree of reaction selectivity. The small pore sizes allow control over which molecules enter the pores, and the flow-through feature can allow for partial oxidation of reactant species as opposed to complete oxidation. A nanoporous separation membrane, produced by ALD is also provided for use in gaseous and liquid separations. The membrane has a high flow rate of material with 100% selectivity. Also provided is a method for producing a catalytic membrane having flow-through pores and discreet catalytic clusters adhering to the inside surfaces of the pores.

  11. Extracorporeal membrane oxygenation circuitry.

    PubMed

    Lequier, Laurance; Horton, Stephen B; McMullan, D Michael; Bartlett, Robert H

    2013-06-01

    The extracorporeal membrane oxygenation circuit is made of a number of components that have been customized to provide adequate tissue oxygen delivery in patients with severe cardiac and/or respiratory failure for a prolonged period of time (days to weeks). A standard extracorporeal membrane oxygenation circuit consists of a mechanical blood pump, gas-exchange device, and a heat exchanger all connected together with circuit tubing. Extracorporeal membrane oxygenation circuits can vary from simple to complex and may include a variety of blood flow and pressure monitors, continuous oxyhemoglobin saturation monitors, circuit access sites, and a bridge connecting the venous access and arterial infusion limbs of the circuit. Significant technical advancements have been made in the equipment available for short- and long-term extracorporeal membrane oxygenation applications. Contemporary extracorporeal membrane oxygenation circuits have greater biocompatibility and allow for more prolonged cardiopulmonary support time while minimizing the procedure-related complications of bleeding, thrombosis, and other physiologic derangements, which were so common with the early application of extracorporeal membrane oxygenation. Modern era extracorporeal membrane oxygenation circuitry and components are simpler, safer, more compact, and can be used across a wide variety of patient sizes from neonates to adults. PMID:23735989

  12. Ordered ceramic membranes

    SciTech Connect

    Anderson, M.A.; Hill, C.G. Jr.; Zeltner, W.A.

    1991-10-01

    Ceramic membranes have been formed from colloidal sols coated on porous clay supports. These supported membranes have been characterized in terms of their permeabilities and permselectivities to various aqueous test solutions. The thermal stabilities and pore structures of these membranes have been characterized by preparing unsupported membranes of the correpsonding material and performing N{sub 2} adsorption-desorption and X-ray diffraction studies on these membranes. To date, membranes have been prepared from a variety of oxides, including TiO{sub 2}, SiO{sub 2}, ZrO{sub 2}, and Al{sub 2}O{sub 3}, as well as Zr-, Fe-, and Nb-doped TiO{sub 2}. In many of these membranes pore diameters are less than 2 nm, while in others the pore diameters are between 3 and 5 nm. Procedures for fabricating porous clay supports with reproducible permeabilities for pure water are also discussed. 30 refs., 59 figs., 22 tabs.

  13. The art of cellular communication: tunneling nanotubes bridge the divide

    PubMed Central

    Gurke, Steffen; Barroso, João F. V.

    2008-01-01

    The ability of cells to receive, process, and respond to information is essential for a variety of biological processes. This is true for the simplest single cell entity as it is for the highly specialized cells of multicellular organisms. In the latter, most cells do not exist as independent units, but are organized into specialized tissues. Within these functional assemblies, cells communicate with each other in different ways to coordinate physiological processes. Recently, a new type of cell-to-cell communication was discovered, based on de novo formation of membranous nanotubes between cells. These F-actin-rich structures, referred to as tunneling nanotubes (TNT), were shown to mediate membrane continuity between connected cells and facilitate the intercellular transport of various cellular components. The subsequent identification of TNT-like structures in numerous cell types revealed some structural diversity. At the same time it emerged that the direct transfer of cargo between cells is a common functional property, suggesting a general role of TNT-like structures in selective, long-range cell-to-cell communication. Due to the growing number of documented thin and long cell protrusions in tissue implicated in cell-to-cell signaling, it is intriguing to speculate that TNT-like structures also exist in vivo and participate in important physiological processes. PMID:18386044

  14. Fendiline inhibits proliferation and invasion of pancreatic cancer cells by interfering with ADAM10 activation and β-catenin signaling

    PubMed Central

    Woods, Neha; Trevino, Jose; Coppola, Domenico; Chellappan, Srikumar; Yang, Shengyu; Padmanabhan, Jaya

    2015-01-01

    ADAM10 (A Disintegrin and Metalloprotease Domain 10) affects the pathophysiology of various cancers, and we had shown that inhibition of ADAM10 sensitizes pancreatic cancer cells to gemcitabine. ADAM10 is activated in response to calcium influx, and here we examined if calcium channel blockers (CCB) would impede ADAM10 activation and affect biology of pancreatic cancer cells. We find that the CCB, fendiline, significantly reduces proliferation, migration, invasion, and anchorage independent growth of pancreatic cancer cells. This was associated with ADAM10 inhibition and its localization at the actin-rich membrane protrusions. Further, fendiline-treated cells formed cadherin-catenin positive tight adherens junctions and elicited defective protein trafficking and recycling. Furthermore, the expression of β-catenin target genes, cyclinD1, c-Myc and CD44, were significantly decreased, suggesting that fendiline might prevent cell proliferation and migration by inhibiting ADAM10 function, cadherin proteolysis and stabilization of cadherin-catenin interaction at the plasma membrane. This will subsequently diminish β-catenin intracellular signaling and repress TCF/LEF target gene expression. Supporting this notion, RNAi-directed downregulation of ADAM10 in cancer cells decreased the expression of cyclinD1, c-Myc and CD44. Furthermore, analysis of human pancreatic tumor tissue microarrays and lysates showed elevated levels of ADAM10, suggesting that aberrant activation of ADAM10 plays a fundamental role in growth and metastasis of PDACs and inhibiting this pathway might be a viable strategy to combat PDACs. PMID:26440150

  15. Palladin interacts with SH3 domains of SPIN90 and Src and is required for Src-induced cytoskeletal remodeling

    PubMed Central

    Rönty, Mikko; Taivainen, Anu; Heiska, Leena; Otey, Carol; Ehler, Elisabeth; Song, Woo Keun; Carpen, Olli

    2007-01-01

    Palladin and SPIN90 are widely expressed proteins, which participate in modulation of actin cytoskeleton by binding to a variety of scaffold and signaling molecules. Cytoskeletal reorganization can induced by activation of signaling pathways, including the PDGF receptor and Src tyrosine kinase pathways. In this study we have analyzed the interplay between palladin, SPIN90 and Src, and characterized the role of palladin and SPIN90 in PDGF and Src-induced cytoskeletal remodeling. We show that the SH3 domains of SPIN90 and Src directly bind palladin’s poly-proline sequence and the interaction controls intracellular targeting of SPIN90. In PDGF-treated cells, palladin and SPIN90 co-localize in actin rich membrane ruffles and lamellipodia. The effect of PDGF on the cytoskeleton is at least partly mediated by the Src kinase, since PP2, a selective Src kinase family inhibitor, blocked PDGF-induced changes. Furthermore, expression of active Src kinase resulted in coordinated translocation of both palladin and SPIN90 to membrane protrusions. Knock-down of endogenous SPIN90 did not inhibit Src-induced cytoskeletal rearrangement, whereas knock-down of palladin resulted in cytoskeletal disorganization and inhibition of remodeling. Further studies showed that palladin is tyrosine phosphorylated in cells expressing active Src indicating bidirectional interplay between palladin and Src. These results may have implications in understanding the invasive and metastatic phenotype of neoplastic cells induced by Src. PMID:17537434

  16. Membrane photobiophysics and photochemistry

    NASA Astrophysics Data System (ADS)

    Ti Tien, H.

    Life, as we know it, depends on solar energy, in particular in the visible range of the solar spectrum. However, visible light alone is useless to the living organism unless a means is available for its capture, transformation, and utilization. Nature, through its long evolution, has perfected a process known as photosynthesis by which visible light is transduced into electrical/ chemical energy. However, the heart of Nature's energy transducer is the thylakoid membrane whose molecular organization was not known until early in the 1960s. Then it was established that the bilayer lipid membrane was central to the design of the thylakoid membrane. To explain the light-driven reactions from water oxidation to carbon dioxide reduction, the so-called Z-scheme was proposed. Concurrent with the establishment of Mitchell's Chemiosmotic Hypothesis for electron transfer and phosphorylation, the experimental bilayer lipid membrane (BLM) system was developed in 1960. But what are the fundamental biophysical mechanisms involved in the phototransduction via pigmented bilayer lipid membrane-based transducers? One of the main purposes of this review is to consider these questions. A second main purpose is to introduce to the reader the experimental aspects of lipid bilayers in the investigation of photoactive biomembranes. The areas covered in this review include a brief summary of the laws of photochemistry relevant to membrane photobiophysics and photobiology. The current exploitation of the BLM system in relation to the thylakoid membrane and to the visual receptor membrane will be considered in turn. The purple membrane of H. Halobium is then discussed. Consideration will also be given to dye-sensitized BLMs, semiconducting BLMs, and BLMs formed from liquid crystals. A common characteristic in the topics covered in this review is the desire to stimulate further studies in the use of the BLM system, not only for the fundamental understanding of biomembranes, but also towards

  17. CENTRIFUGAL MEMBRANE FILTRATION

    SciTech Connect

    Daniel J. Stepan; Bradley G. Stevens; Melanie D. Hetland

    1999-10-01

    The overall project consists of several integrated research phases related to the applicability, continued development, demonstration, and commercialization of the SpinTek centrifugal membrane filtration process. Work performed during this reporting period consisted of Phase 2 evaluation of the SpinTek centrifugal membrane filtration technology and Phase 3, Technology Partnering. During Phase 1 testing conducted at the EERC using the SpinTek ST-IIL unit operating on a surrogate tank waste, a solids cake developed on the membrane surface. The solids cake was observed where linear membrane velocities were less than 17.5 ft/s and reduced the unobstructed membrane surface area up to 25%, reducing overall filtration performance. The primary goal of the Phase 2 research effort was to enhance filtration performance through the development and testing of alternative turbulence promoter designs. The turbulence promoters were designed to generate a shear force across the entire membrane surface sufficient to maintain a self-cleaning membrane capability and improve filtration efficiency and long-term performance. Specific Phase 2 research activities included the following: System modifications to accommodate an 11-in.-diameter, two-disk rotating membrane assembly; Development and fabrication of alternative turbulence promoter designs; Testing and evaluation of the existing and alternative turbulence promoters under selected operating conditions using a statistically designed test matrix; and Data reduction and analysis; The objective of Phase 3 research was to demonstrate the effectiveness of SpinTek's centrifugal membrane filtration as a pretreatment to remove suspended solids from a liquid waste upstream of 3M's WWL cartridge technology for the selective removal of technetium (Tc).

  18. Nanoengineered membrane electrode assembly interface

    DOEpatents

    Song, Yujiang; Shelnutt, John A

    2013-08-06

    A membrane electrode structure suitable for use in a membrane electrode assembly (MEA) that comprises membrane-affixed metal nanoparticles whose formation is controlled by a photochemical process that controls deposition of the metal nanoparticles using a photocatalyst integrated with a polymer electrolyte membrane, such as an ionomer membrane. Impregnation of the polymer membrane with the photocatalyst prior to metal deposition greatly reduces the required amount of metal precursor in the deposition reaction solution by restricting metal reduction substantially to the formation of metal nanoparticles affixed on or near the surface of the polymer membrane with minimal formation of metallic particles not directly associated with the membrane.

  19. Building interconnected membrane networks.

    PubMed

    Holden, Matthew A

    2015-01-01

    Reconstituted replica cell membranes are easily created by contacting two lipid-monolayer-encased aqueous droplets under an oil phase. Called the droplet interface bilayer (DIB), this technique has been used to study a wide range of membrane processes. Importantly, this method is compatible with electrical measurements, meaning that membrane protein activities are easily observed in DIBs. By positioning droplets in two- and three-dimensional networks, sophisticated interconnected systems can be created that possess collective properties. The methods described here summarize the approaches used to create DIB networks and how to operate the devices that have been constructed so far.

  20. Rotating bubble membrane radiator

    DOEpatents

    Webb, Brent J.; Coomes, Edmund P.

    1988-12-06

    A heat radiator useful for expelling waste heat from a power generating system aboard a space vehicle is disclosed. Liquid to be cooled is passed to the interior of a rotating bubble membrane radiator, where it is sprayed into the interior of the bubble. Liquid impacting upon the interior surface of the bubble is cooled and the heat radiated from the outer surface of the membrane. Cooled liquid is collected by the action of centrifical force about the equator of the rotating membrane and returned to the power system. Details regarding a complete space power system employing the radiator are given.

  1. Neural Membrane Signaling Platforms

    PubMed Central

    Wallace, Ron

    2010-01-01

    Throughout much of the history of biology, the cell membrane was functionally defined as a semi-permeable barrier separating aqueous compartments, and an anchoring site for proteins. Little attention was devoted to its possible regulatory role in intracellular molecular processes and neuron electrical signaling. This article reviews the history of membrane studies and the current state of the art. Emphasis is placed on natural and artificial membrane studies of electric field effects on molecular organization, especially as these may relate to impulse propagation in neurons. Implications of these studies for new designs in artificial intelligence are briefly examined. PMID:20640161

  2. Cyclic membrane separation process

    DOEpatents

    Nemser, Stuart M.

    2005-05-03

    A cyclic process for controlling environmental emissions of volatile organic compounds (VOC) from vapor recovery in storage and dispensing operations of liquids maintains a vacuum in the storage tank ullage. In the first part of a two-part cyclic process ullage vapor is discharged through a vapor recovery system in which VOC are stripped from vented gas with a selectively gas permeable membrane. In the second part, the membrane is inoperative while gas pressure rises in the ullage. In one aspect of this invention, a vacuum is drawn in the membrane separation unit thus reducing overall VOC emissions.

  3. Composite metal membrane

    DOEpatents

    Peachey, N.M.; Dye, R.C.; Snow, R.C.; Birdsell, S.A.

    1998-04-14

    A composite metal membrane including a first metal layer of Group IVB met or Group VB metals, the first metal layer sandwiched between two layers of an oriented metal of palladium, platinum or alloys thereof is provided together with a process for the recovery of hydrogen from a gaseous mixture including contacting a hydrogen-containing gaseous mixture with a first side of a nonporous composite metal membrane including a first metal of Group IVB metals or Group VB metals, the first metal layer sandwiched between two layers of an oriented metal of palladium, platinum or alloys thereof, and, separating hydrogen from a second side of the nonporous composite metal membrane.

  4. Composite metal membrane

    DOEpatents

    Peachey, Nathaniel M.; Dye, Robert C.; Snow, Ronny C.; Birdsell, Stephan A.

    1998-01-01

    A composite metal membrane including a first metal layer of Group IVB met or Group VB metals, the first metal layer sandwiched between two layers of an oriented metal of palladium, platinum or alloys thereof is provided together with a process for the recovery of hydrogen from a gaseous mixture including contacting a hydrogen-containing gaseous mixture with a first side of a nonporous composite metal membrane including a first metal of Group IVB metals or Group VB metals, the first metal layer sandwiched between two layers of an oriented metal of palladium, platinum or alloys thereof, and, separating hydrogen from a second side of the nonporous composite metal membrane.

  5. A computer simulation approach to quantify the true area and true area compressibility modulus of biological membranes

    SciTech Connect

    Chacón, Enrique; Tarazona, Pedro; Bresme, Fernando

    2015-07-21

    We present a new computational approach to quantify the area per lipid and the area compressibility modulus of biological membranes. Our method relies on the analysis of the membrane fluctuations using our recently introduced coupled undulatory (CU) mode [Tarazona et al., J. Chem. Phys. 139, 094902 (2013)], which provides excellent estimates of the bending modulus of model membranes. Unlike the projected area, widely used in computer simulations of membranes, the CU area is thermodynamically consistent. This new area definition makes it possible to accurately estimate the area of the undulating bilayer, and the area per lipid, by excluding any contributions related to the phospholipid protrusions. We find that the area per phospholipid and the area compressibility modulus features a negligible dependence with system size, making possible their computation using truly small bilayers, involving a few hundred lipids. The area compressibility modulus obtained from the analysis of the CU area fluctuations is fully consistent with the Hooke’s law route. Unlike existing methods, our approach relies on a single simulation, and no a priori knowledge of the bending modulus is required. We illustrate our method by analyzing 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine bilayers using the coarse grained MARTINI force-field. The area per lipid and area compressibility modulus obtained with our method and the MARTINI forcefield are consistent with previous studies of these bilayers.

  6. A computer simulation approach to quantify the true area and true area compressibility modulus of biological membranes

    NASA Astrophysics Data System (ADS)

    Chacón, Enrique; Tarazona, Pedro; Bresme, Fernando

    2015-07-01

    We present a new computational approach to quantify the area per lipid and the area compressibility modulus of biological membranes. Our method relies on the analysis of the membrane fluctuations using our recently introduced coupled undulatory (CU) mode [Tarazona et al., J. Chem. Phys. 139, 094902 (2013)], which provides excellent estimates of the bending modulus of model membranes. Unlike the projected area, widely used in computer simulations of membranes, the CU area is thermodynamically consistent. This new area definition makes it possible to accurately estimate the area of the undulating bilayer, and the area per lipid, by excluding any contributions related to the phospholipid protrusions. We find that the area per phospholipid and the area compressibility modulus features a negligible dependence with system size, making possible their computation using truly small bilayers, involving a few hundred lipids. The area compressibility modulus obtained from the analysis of the CU area fluctuations is fully consistent with the Hooke's law route. Unlike existing methods, our approach relies on a single simulation, and no a priori knowledge of the bending modulus is required. We illustrate our method by analyzing 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine bilayers using the coarse grained MARTINI force-field. The area per lipid and area compressibility modulus obtained with our method and the MARTINI forcefield are consistent with previous studies of these bilayers.

  7. Metabolic regulation of neutrophil spreading, membrane tubulovesicular extensions (cytonemes) formation and intracellular pH upon adhesion to fibronectin

    SciTech Connect

    Galkina, Svetlana I. . E-mail: galkina@genebee.msu.su; Sud'ina, Galina F.; Klein, Thomas

    2006-08-01

    Circulating leukocytes have a round cell shape and roll along vessel walls. However, metabolic disorders can lead them to adhere to the endothelium and spread (flatten). We studied the metabolic regulation of adhesion, spreading and intracellular pH (pHi) of neutrophils (polymorphonuclear leukocytes) upon adhesion to fibronectin-coated substrata. Resting neutrophils adhered and spread on fibronectin. An increase in pHi accompanied neutrophil spreading. Inhibition of oxidative phosphorylation or inhibition of P- and F-type ATPases affected neither neutrophil spreading nor pHi. Inhibition of glucose metabolism or V-ATPase impaired neutrophil spreading, blocked the increase in the pHi and induced extrusion of membrane tubulovesicular extensions (cytonemes), anchoring cells to substrata. Omission of extracellular Na{sup +} and inhibition of chloride channels caused a similar effect. We propose that these tubulovesicular extensions represent protrusions of exocytotic trafficking, supplying the plasma membrane of neutrophils with ion exchange mechanisms and additional membrane for spreading. Glucose metabolism and V-type ATPase could affect fusion of exocytotic trafficking with the plasma membrane, thus controlling neutrophil adhesive state and pHi. Cl{sup -} efflux through chloride channels and Na{sup +} influx seem to be involved in the regulation of the V-ATPase by carrying out charge compensation for the proton-pumping activity and through V-ATPase in regulation of neutrophil spreading and pHi.

  8. Fuel cell membrane humidification

    DOEpatents

    Wilson, Mahlon S.

    1999-01-01

    A polymer electrolyte membrane fuel cell assembly has an anode side and a cathode side separated by the membrane and generating electrical current by electrochemical reactions between a fuel gas and an oxidant. The anode side comprises a hydrophobic gas diffusion backing contacting one side of the membrane and having hydrophilic areas therein for providing liquid water directly to the one side of the membrane through the hydrophilic areas of the gas diffusion backing. In a preferred embodiment, the hydrophilic areas of the gas diffusion backing are formed by sewing a hydrophilic thread through the backing. Liquid water is distributed over the gas diffusion backing in distribution channels that are separate from the fuel distribution channels.

  9. Evaporative Cooling Membrane Device

    NASA Technical Reports Server (NTRS)

    Lomax, Curtis (Inventor); Moskito, John (Inventor)

    1999-01-01

    An evaporative cooling membrane device is disclosed having a flat or pleated plate housing with an enclosed bottom and an exposed top that is covered with at least one sheet of hydrophobic porous material having a thin thickness so as to serve as a membrane. The hydrophobic porous material has pores with predetermined dimensions so as to resist any fluid in its liquid state from passing therethrough but to allow passage of the fluid in its vapor state, thereby, causing the evaporation of the fluid and the cooling of the remaining fluid. The fluid has a predetermined flow rate. The evaporative cooling membrane device has a channel which is sized in cooperation with the predetermined flow rate of the fluid so as to produce laminar flow therein. The evaporative cooling membrane device provides for the convenient control of the evaporation rates of the circulating fluid by adjusting the flow rates of the laminar flowing fluid.

  10. Hydrogen transport membranes

    DOEpatents

    Mundschau, Michael V.

    2005-05-31

    Composite hydrogen transport membranes, which are used for extraction of hydrogen from gas mixtures are provided. Methods are described for supporting metals and metal alloys which have high hydrogen permeability, but which are either too thin to be self supporting, too weak to resist differential pressures across the membrane, or which become embrittled by hydrogen. Support materials are chosen to be lattice matched to the metals and metal alloys. Preferred metals with high permeability for hydrogen include vanadium, niobium, tantalum, zirconium, palladium, and alloys thereof. Hydrogen-permeable membranes include those in which the pores of a porous support matrix are blocked by hydrogen-permeable metals and metal alloys, those in which the pores of a porous metal matrix are blocked with materials which make the membrane impervious to gases other than hydrogen, and cermets fabricated by sintering powders of metals with powders of lattice-matched ceramic.

  11. Structures of membrane proteins

    PubMed Central

    Vinothkumar, Kutti R.; Henderson, Richard

    2010-01-01

    In reviewing the structures of membrane proteins determined up to the end of 2009, we present in words and pictures the most informative examples from each family. We group the structures together according to their function and architecture to provide an overview of the major principles and variations on the most common themes. The first structures, determined 20 years ago, were those of naturally abundant proteins with limited conformational variability, and each membrane protein structure determined was a major landmark. With the advent of complete genome sequences and efficient expression systems, there has been an explosion in the rate of membrane protein structure determination, with many classes represented. New structures are published every month and more than 150 unique membrane protein structures have been determined. This review analyses the reasons for this success, discusses the challenges that still lie ahead, and presents a concise summary of the key achievements with illustrated examples selected from each class. PMID:20667175

  12. Novel Catalytic Membrane Reactors

    SciTech Connect

    2009-02-01

    This factsheet describes a research project that will focus on the development and application of nonporous high gas flux perfluoro membranes with high temperature rating and excellent chemical resistance.

  13. Drugging Membrane Protein Interactions

    PubMed Central

    Yin, Hang; Flynn, Aaron D.

    2016-01-01

    The majority of therapeutics target membrane proteins, accessible on the surface of cells, to alter cellular signaling. Cells use membrane proteins to transduce signals into cells, transport ions and molecules, bind the cell to a surface or substrate, and catalyze reactions. Newly devised technologies allow us to drug conventionally “undruggable” regions of membrane proteins, enabling modulation of protein–protein, protein–lipid, and protein–nucleic acid interactions. In this review, we survey the state of the art in high-throughput screening and rational design in drug discovery, and we evaluate the advances in biological understanding and technological capacity that will drive pharmacotherapy forward against unorthodox membrane protein targets. PMID:26863923

  14. Membrane separation of hydrocarbons

    DOEpatents

    Funk, Edward W.; Kulkarni, Sudhir S.; Chang, Y. Alice

    1986-01-01

    Mixtures of heavy oils and light hydrocarbons may be separated by passing the mixture over a polymeric membrane which comprises a polymer capable of maintaining its integrity in the presence of hydrocarbon compounds at temperature ranging from about ambient to about 100.degree. C. and pressures ranging from about 50 to about 1000 psi. The membranes which possess pore sizes ranging from about 10 to about 500 Angstroms are cast from a solvent solution and recovered.

  15. Battery utilizing ceramic membranes

    DOEpatents

    Yahnke, Mark S.; Shlomo, Golan; Anderson, Marc A.

    1994-01-01

    A thin film battery is disclosed based on the use of ceramic membrane technology. The battery includes a pair of conductive collectors on which the materials for the anode and the cathode may be spin coated. The separator is formed of a porous metal oxide ceramic membrane impregnated with electrolyte so that electrical separation is maintained while ion mobility is also maintained. The entire battery can be made less than 10 microns thick while generating a potential in the 1 volt range.

  16. [Candidiasis of mucous membranes].

    PubMed

    Khmel'nitskiĭ, O K

    2000-01-01

    The author presents a new concept of complex relationship between the organism and Candida. According to this concept there is transformation of Candida carriage into invasive candidiasis of the mucous membranes. The idea is formulated on mixed-mycocenosis as association of fungal, bacterial, protozoic and viral biota. For the first time the notion of intermediate preinvasive form of candidiasis is given and clinical, morphological criteria of differential diagnosis between candida carriage, preinvasive and invasive candidiasis of mucous membranes are presented. PMID:11198119

  17. Microprobes aluminosilicate ceramic membranes

    DOEpatents

    Anderson, Marc A.; Sheng, Guangyao

    1993-01-01

    Methods have been developed to make mixed alumina-silicate and aluminosilicate particulate microporous ceramic membranes. One method involves the making of separate alumina and silica sols which are then mixed. Another method involves the creation of a combined sol with aluminosilicate particles. The resulting combined alumina and silica membranes have high surface area, a very small pore size, and a very good temperature stability.

  18. Membrane reference electrode

    DOEpatents

    Redey, L.; Bloom, I.D.

    1988-01-21

    A reference electrode utilizes a small thin, flat membrane of a highly conductive glass placed on a small diameter insulator tube having a reference material inside in contact with an internal voltage lead. When the sensor is placed in a non-aqueous ionic electrolytic solution, the concentration difference across the glass membrane generates a low voltage signal in precise relationship to the concentration of the species to be measured, with high spatial resolution. 2 figs.

  19. Inverse colloidal crystal membranes for hydrophobic interaction membrane chromatography.

    PubMed

    Vu, Anh T; Wang, Xinying; Wickramasinghe, S Ranil; Yu, Bing; Yuan, Hua; Cong, Hailin; Luo, Yongli; Tang, Jianguo

    2015-08-01

    Hydrophobic interaction membrane chromatography has gained interest due to its excellent performance in the purification of humanized monoclonal antibodies. The membrane material used in hydrophobic interaction membrane chromatography has typically been commercially available polyvinylidene fluoride. In this contribution, newly developed inverse colloidal crystal membranes that have uniform pores, high porosity and, therefore, high surface area for protein binding are used as hydrophobic interaction membrane chromatography membranes for humanized monoclonal antibody immunoglobulin G purification. The capacity of the inverse colloidal crystal membranes developed here is up to ten times greater than commercially available polyvinylidene fluoride membranes with a similar pore size. This work highlights the importance of developing uniform pore size high porosity membranes in order to maximize the capacity of hydrophobic interaction membrane chromatography.

  20. Catalytic nanoporous membranes

    DOEpatents

    Pellin, Michael J.; Hryn, John N.; Elam, Jeffrey W.

    2009-12-01

    A nanoporous catalytic membrane which displays several unique features including pores which can go through the entire thickness of the membrane. The membrane has a higher catalytic and product selectivity than conventional catalysts. Anodic aluminum oxide (AAO) membranes serve as the catalyst substrate. This substrate is then subjected to Atomic Layer Deposition (ALD), which allows the controlled narrowing of the pores from 40 nm to 10 nm in the substrate by deposition of a preparatory material. Subsequent deposition of a catalytic layer on the inner surfaces of the pores reduces pore sizes to less than 10 nm and allows for a higher degree of reaction selectivity. The small pore sizes allow control over which molecules enter the pores, and the flow-through feature can allow for partial oxidation of reactant species as opposed to complete oxidation. A nanoporous separation membrane, produced by ALD is also provided for use in gaseous and liquid separations. The membrane has a high flow rate of material with 100% selectivity.

  1. Criticality in Plasma Membranes

    NASA Astrophysics Data System (ADS)

    Machta, Benjamin; Papanikolaou, Stefanos; Sethna, James; Veatch, Sarah

    2011-03-01

    We are motivated by recent observations of micron-sized critical fluctuations in the 2d Ising Universality class in plasma membrane vesicles that are isolated from cortical cytoskeleton. We construct a minimal model of the plasma membrane's interaction with intact cytoskeleton which explains why large scale phase separation has not been observed in Vivo. In addition, we use analytical techniques from conformal field theory and numerical simulations to investigate the form of effective forces mediated by the membrane's proximity to criticality. We show that the range of this force is maximized near a critical point and we quantify its usefulness in mediating communication using techniques from information theory. Finally we use theoretical techniques from statistical physics in conjunction with Monte-Carlo simulations to understand how criticality can be used to increase the efficiency of membrane bound receptor mediated signaling. We expect that this sort of analysis will be broadly useful in understanding and quantifying the role of lipid ``rafts'' in a wide variety of membrane bound processes. Generally, we demonstrate that critical fluctuations provide a physical mechanism to organize and spatially segregate membrane components by providing channels for interaction over relatively large distances.

  2. OXYGEN TRANSPORT CERAMIC MEMBRANES

    SciTech Connect

    Dr. Sukumar Bandopadhyay; Dr. Nagendra Nagabhushana

    2000-10-01

    This is the third quarterly report on oxygen Transport Ceramic Membranes. In the following, the report describes the progress made by our university partners in Tasks 1 through 6, experimental apparatus that was designed and built for various tasks of this project, thermodynamic calculations, where applicable and work planned for the future. (Task 1) Design, fabricate and evaluate ceramic to metal seals based on graded ceramic powder/metal braze joints. (Task 2) Evaluate the effect of defect configuration on ceramic membrane conductivity and long term chemical and structural stability. (Task 3) Determine materials mechanical properties under conditions of high temperatures and reactive atmospheres. (Task 4) Evaluate phase stability and thermal expansion of candidate perovskite membranes and develop techniques to support these materials on porous metal structures. (Task 5) Assess the microstructure of membrane materials to evaluate the effects of vacancy-impurity association, defect clusters, and vacancy-dopant association on the membrane performance and stability. (Task 6) Measure kinetics of oxygen uptake and transport in ceramic membrane materials under commercially relevant conditions using isotope labeling techniques.

  3. Fractal reconstruction of rough membrane surface related with membrane fouling in a membrane bioreactor.

    PubMed

    Zhang, Meijia; Chen, Jianrong; Ma, Yuanjun; Shen, Liguo; He, Yiming; Lin, Hongjun

    2016-09-01

    In this paper, fractal reconstruction of rough membrane surface with a modified Weierstrass-Mandelbrot (WM) function was conducted. The topography of rough membrane surface was measured by an atomic force microscopy (AFM), and the results showed that the membrane surface was isotropous. Accordingly, the fractal dimension and roughness of membrane surface were calculated by the power spectrum method. The rough membrane surface was reconstructed on the MATLAB platform with the parameter values acquired from raw AFM data. The reconstructed membrane was much similar to the real membrane morphology measured by AFM. The parameters (including average roughness and root mean square (RMS) roughness) associated with membrane morphology for the model and real membrane were calculated, and a good match of roughness parameters between the reconstructed surface and real membrane was found, indicating the feasibility of the new developed method. The reconstructed membrane surface can be potentially used for interaction energy evaluation. PMID:27318159

  4. Fractal reconstruction of rough membrane surface related with membrane fouling in a membrane bioreactor.

    PubMed

    Zhang, Meijia; Chen, Jianrong; Ma, Yuanjun; Shen, Liguo; He, Yiming; Lin, Hongjun

    2016-09-01

    In this paper, fractal reconstruction of rough membrane surface with a modified Weierstrass-Mandelbrot (WM) function was conducted. The topography of rough membrane surface was measured by an atomic force microscopy (AFM), and the results showed that the membrane surface was isotropous. Accordingly, the fractal dimension and roughness of membrane surface were calculated by the power spectrum method. The rough membrane surface was reconstructed on the MATLAB platform with the parameter values acquired from raw AFM data. The reconstructed membrane was much similar to the real membrane morphology measured by AFM. The parameters (including average roughness and root mean square (RMS) roughness) associated with membrane morphology for the model and real membrane were calculated, and a good match of roughness parameters between the reconstructed surface and real membrane was found, indicating the feasibility of the new developed method. The reconstructed membrane surface can be potentially used for interaction energy evaluation.

  5. Three-dimensional in vivo analysis of Dictyostelium mounds reveals directional sorting of prestalk cells and defines a role for the myosin II regulatory light chain in prestalk cell sorting and tip protrusion.

    PubMed

    Clow, P A; Chen, T; Chisholm, R L; McNally, J G

    2000-06-01

    During cell sorting in Dictyostelium, we observed that GFP-tagged prestalk cells (ecmAO-expressing cells) moved independently and directionally to form a cluster. This is consistent with a chemotaxis model for cell sorting (and not differential adhesion) in which a long-range signal attracts many of the prestalk cells to the site of cluster formation. Surprisingly, the ecmAO prestalk cluster that we observed was initially found at a random location within the mound of this Ax3 strain, defining an intermediate sorting stage not widely reported in Dictyostelium. The cluster then moved en masse to the top of the mound to produce the classic, apical pattern of ecmAO prestalk cells. Migration of the cluster was also directional, suggesting the presence of another long-range guidance cue. Once at the mound apex, the cluster continued moving upward leading to protrusion of the mound's tip. To investigate the role of the cluster in tip protrusion, we examined ecmAO prestalk-cell sorting in a myosin II regulatory light chain (RLC) null in which tips fail to form. In RLC-null mounds, ecmAO prestalk cells formed an initial cluster that began to move to the mound apex, but then arrested as a vertical column that extended from the mound's apex to its base. Mixing experiments with wild-type cells demonstrated that the RLC-null ecmAO prestalk-cell defect is cell autonomous. These observations define a specific mechanism for myosin's function in tip formation, namely a mechanical role in the upward movement of the ecmAO prestalk cluster. The wild-type data demonstrate that cell sorting can occur in two steps, suggesting that, in this Ax3 strain, spatially and temporally distinct cues may guide prestalk cells first to an initial cluster and then later to the tip.

  6. Composite membranes for fluid separations

    DOEpatents

    Blume, Ingo; Peinemann, Klaus-Viktor; Pinnau, Ingo; Wijmans, Johannes G.

    1992-01-01

    A method for designing and making composite membranes having a microporous support membrane coated with a permselective layer. The method involves calculating the minimum thickness of the permselective layer such that the selectivity of the composite membrane is close to the intrinsic selectivity of the perselective layer. The invention also provides high performance membranes with optimized properties.

  7. Composite membranes for fluid separations

    DOEpatents

    Blume, Ingo; Peinemann, Klaus-Viktor; Pinnau, Ingo; Wijmans, Johannes G.

    1990-01-01

    A method for designing and making composite membranes having a microporous support membrane coated with a permselective layer. The method involves calculating the minimum thickness of the permselective layer such that the selectivity of the composite membrane is close to the intrinsic selectivity of the permselective layer. The invention also provides high performance membranes with optimized properties.

  8. Composite membranes for fluid separations

    DOEpatents

    Blume, Ingo; Peinemann, Klaus-Viktor; Pinnau, Ingo; Wijmans, Johannes G.

    1991-01-01

    A method for designing and making composite membranes having a microporous support membrane coated with a permselective layer. The method involves calculating the minimum thickness of the permselective layer such that the selectivity of the composite membrane is close to the intrinsic selectivity of the permselective layer. The invention also provides high performance membranes with optimized properties.

  9. Asymmetric membranes for gas separations

    SciTech Connect

    Finken, H.

    1985-01-01

    Recent membrane developments for gaseous mixture separations are compared to the development of reverse osmosis membranes for water desalination. The goals of these developments have been the search for ideal permselective polymeric materials, techniques for producing ultrathin membrane layers free of imperfections and transforming gelled reverse osmosis membranes into solid gas permeation membranes. A novel approach to meeting the basic requirements of high permselectivity is attempted by altering the physical polymer structure within the membrane prior to application for gas separation. The influence of these physical interactions on membrane properties is presented. 47 references, 11 figures, 6 tables.

  10. Oxygen Transport Ceramic Membranes

    SciTech Connect

    S. Bandopadhyay; T. Nithyanantham; X.-D Zhou; Y-W. Sin; H.U. Anderson; Alan Jacobson; C.A. Mims

    2005-11-01

    The present quarterly report describes some of the investigations on the structural properties of dense OTM bars provided by Praxair and studies on newer composition of Ti doped LSF. In the current research, the electrical conductivity and Seebeck coefficient were measured as a function of temperature in air. Based on these measurements, the charge carrier concentration, net acceptor dopant concentration, activation energy of conduction and mobility were estimated. The studies on the fracture toughness of the LSFT and dual phase membranes at room temperature have been completed and reported previously. The membranes that are exposed to high temperatures at an inert and a reactive atmosphere undergo many structural and chemical changes which affects the mechanical properties. To study the effect of temperature on the membranes when exposed to an inert environment, the membranes (LAFT and Dual phase) were heat treated at 1000 C in air and N{sub 2} atmosphere and hardness and fracture toughness of the membranes were studied after the treatment. The indentation method was used to find the fracture toughness and the effect of the heat treatment on the mechanical properties of the membranes. Further results on the investigation of the origin of the slow kinetics on reduction of ferrites have been obtained. The slow kinetics appears to be related to a non-equilibrium reduction pathway that initially results in the formation of iron particles. At long times, equilibrium can be reestablished with recovery of the perovskite phase. 2-D modeling of oxygen movement has been undertaken in order to fit isotope data. The model will serve to study ''frozen'' profiles in patterned or composite membranes.

  11. Denitrification incorporating microporous membranes

    SciTech Connect

    Reising, A.R.; Schroeder, E.D.

    1996-07-01

    A microbial system for removal of nitrate from drinking water in which the denitrification reactions are physically separated from the water being treated by a microporous membrane was studied. The experimental system was composed of two equal-volume cells separated by a 0.02 {mu}m pore size polytetrafluoroethylene membrane. No pressure difference existed across the membrane, and nitrate ions were transported through the membrane by diffusion. Results of experiments with biofilm denitrification and suspended-culture systems are presented. Deoxygenation by N{sub 2} stripping and by SO{sub 3}{sup {minus}2} produced equivalent results. Removal rates with the suspended culture systems were greater than those with biofilms. Effective membrane diffusivities calculated for suspended-culture systems were approximately the same as for sterile systems, indicating that resistance to transport was not changed by the addition of the suspended microbial culture. The methanol requirement observed in the experiments was 1.4 g total organic carbon (TOC) per g NO{sub 3}{sup {minus}}-N removed. Removal rates followed first-order pseudotransport controlled models for both the biofilm and suspended-culture systems. A prototype continuous-flow system is presented.

  12. Micron-scale plasma membrane curvature is recognized by the septin cytoskeleton

    PubMed Central

    Bridges, Andrew A.; Jentzsch, Maximilian S.; Oakes, Patrick W.; Occhipinti, Patricia

    2016-01-01

    Cells change shape in response to diverse environmental and developmental conditions, creating topologies with micron-scale features. Although individual proteins can sense nanometer-scale membrane curvature, it is unclear if a cell could also use nanometer-scale components to sense micron-scale contours, such as the cytokinetic furrow and base of neuronal branches. Septins are filament-forming proteins that serve as signaling platforms and are frequently associated with areas of the plasma membrane where there is micron-scale curvature, including the cytokinetic furrow and the base of cell protrusions. We report here that fungal and human septins are able to distinguish between different degrees of micron-scale curvature in cells. By preparing supported lipid bilayers on beads of different curvature, we reconstitute and measure the intrinsic septin curvature preference. We conclude that micron-scale curvature recognition is a fundamental property of the septin cytoskeleton that provides the cell with a mechanism to know its local shape. PMID:27044896

  13. Lidocaine induces ROCK-dependent membrane blebbing and subsequent cell death in rabbit articular chondrocytes.

    PubMed

    Maeda, Tsutomu; Toyoda, Futoshi; Imai, Shinji; Tanigawa, Hitoshi; Kumagai, Kousuke; Matsuura, Hiroshi; Matsusue, Yoshitaka

    2016-05-01

    Local anesthetics are administered intraarticularly for pain control in orthopedic clinics and surgeries. Although previous studies have shown that local anesthetics can be toxic to chondrocytes, the underlying cellular mechanisms remain unclear. The present study investigates acute cellular responses associated with lidocaine-induced toxicity to articular chondrocytes. Rabbit articular chondrocytes were exposed to lidocaine and their morphological changes were monitored with live cell microscopy. The viability of chondrocytes was evaluated using a fluorescence based LIVE/DEAD assay. Acute treatment of chondrocytes with lidocaine (3-30 mM) induced spherical protrusions on the cell surface (so called "membrane blebbing") in a time- and concentration-dependent manner. The concentration-response relationship for the lidocaine effect was shifted leftward by elevating extracellular pH, as expected for the non-ionized lidocaine being involved in the bleb formation. ROCK (Rho-kinase) inhibitors Y-27632 and fasudil completely prevented the lidocaine-induced membrane blebbing, suggesting that ROCK activation is required for bleb formation. Caspase-3 levels were unchanged by 10 mM lidocaine (p = 0.325) and a caspase inhibitor z-VAD-fmk did not affect the lidocaine-induced blebbing (p = 0.964). GTP-RhoA levels were significantly increased (p < 0.001), but Rho inhibitor-1 failed to suppress the membrane blebbing (p = 0.875). Lidocaine (30 mM) reduced the cell viability of isolated chondrocytes (p < 0.001) and in situ chondrocytes (p < 0.001). The chondrotoxicity was attenuated by pretreatment of cells with ROCK inhibitors or a myosin-II inhibitor blebbistatin (p < 0.001). These findings suggest that lidocaine induces ROCK-dependent membrane blebbing and thereby produces a cytotoxic effect on chondrocytes. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:754-762, 2016.

  14. Oxygen Transport Ceramic Membranes

    SciTech Connect

    S. Bandopadhyay; T. Nithyanantham; X.-D Zhou; Y-W. Sin; H.U. Anderson; Alan Jacobson; C.A. Mims

    2006-05-01

    In this quarter a systematic analysis on the decomposition behavior of the OTM membranes at air and nitrogen were initiated to understand the structural and stoichiometric changes associated with elevated temperatures. Evaluation of the flexural strengths using 4-point bend test was also started for the dual phase membranes. Initial results on the synthesis of dual phase composite materials have been obtained. The measurements have focused on the compatibility of mixed conductors with the pure ionic conductors yttria stabilized zirconia (YSZ) and gadolinium doped ceria (GDC). The initial results obtained for three different mixed conductors suggest that (GDC) is the better choice. A new membrane permeation system has been designed and tested and sintering studies of biphasic systems are in progress.

  15. Viral membrane fusion.

    PubMed

    Harrison, Stephen C

    2015-05-01

    Membrane fusion is an essential step when enveloped viruses enter cells. Lipid bilayer fusion requires catalysis to overcome a high kinetic barrier; viral fusion proteins are the agents that fulfill this catalytic function. Despite a variety of molecular architectures, these proteins facilitate fusion by essentially the same generic mechanism. Stimulated by a signal associated with arrival at the cell to be infected (e.g., receptor or co-receptor binding, proton binding in an endosome), they undergo a series of conformational changes. A hydrophobic segment (a "fusion loop" or "fusion peptide") engages the target-cell membrane and collapse of the bridging intermediate thus formed draws the two membranes (virus and cell) together. We know of three structural classes for viral fusion proteins. Structures for both pre- and postfusion conformations of illustrate the beginning and end points of a process that can be probed by single-virion measurements of fusion kinetics.

  16. Altering morphology of membranes

    SciTech Connect

    Narayan, R.S.

    1987-01-01

    The use of membranes in industrial gas processing and separation has been on the increase in the last eight years, especially since the successful introduction and commercialization of PRISM separators by Monsanto in 1979. Since that time, a number of gas separation applications have been successfully applied on an industrial scale. Separation of hydrogen from N/sub 2/ and hydrocarbons using membranes have become commonplace, and separation of carbon dioxide from hydrocarbons is gaining increasing acceptance. More recently, the separation of nitrogen from oxygen from air has been gaining considerable attention. The economic benefits of using membranes for on-site generation of N/sub 2/ containing less than 5% O/sub 2/ for inert blanketing such as, product storage, packaging, on board ships, airplanes and oil/gas production platforms, etc., are demonstrated to be significant.

  17. Hybrid Filter Membrane

    NASA Technical Reports Server (NTRS)

    Laicer, Castro; Rasimick, Brian; Green, Zachary

    2012-01-01

    Cabin environmental control is an important issue for a successful Moon mission. Due to the unique environment of the Moon, lunar dust control is one of the main problems that significantly diminishes the air quality inside spacecraft cabins. Therefore, this innovation was motivated by NASA s need to minimize the negative health impact that air-suspended lunar dust particles have on astronauts in spacecraft cabins. It is based on fabrication of a hybrid filter comprising nanofiber nonwoven layers coated on porous polymer membranes with uniform cylindrical pores. This design results in a high-efficiency gas particulate filter with low pressure drop and the ability to be easily regenerated to restore filtration performance. A hybrid filter was developed consisting of a porous membrane with uniform, micron-sized, cylindrical pore channels coated with a thin nanofiber layer. Compared to conventional filter media such as a high-efficiency particulate air (HEPA) filter, this filter is designed to provide high particle efficiency, low pressure drop, and the ability to be regenerated. These membranes have well-defined micron-sized pores and can be used independently as air filters with discreet particle size cut-off, or coated with nanofiber layers for filtration of ultrafine nanoscale particles. The filter consists of a thin design intended to facilitate filter regeneration by localized air pulsing. The two main features of this invention are the concept of combining a micro-engineered straight-pore membrane with nanofibers. The micro-engineered straight pore membrane can be prepared with extremely high precision. Because the resulting membrane pores are straight and not tortuous like those found in conventional filters, the pressure drop across the filter is significantly reduced. The nanofiber layer is applied as a very thin coating to enhance filtration efficiency for fine nanoscale particles. Additionally, the thin nanofiber coating is designed to promote capture of

  18. Functional membranes. Present and future

    NASA Technical Reports Server (NTRS)

    Kunitake, T.

    1982-01-01

    The present situation and the future development of the functional membrane are discussed. It is expected that functional membranes will play increasingly greater roles in the chemical industry of the coming decade. These membranes are formed from polymer films, liquid membranes or bilayer membranes. The two most important technologies based on the polymeric membrane are reverse osmosis and ion exchange. The liquid membrane is used for separation of ionic species; an extension of the solvent extraction process. By using appropriate ligands and ionophores, highly selective separations are realized. The active transport is made possible if the physical and chemical potentials are applied to the transport process. More advanced functional membranes may be designed on the basis of the synthetic bilayer membrane.

  19. Polystyrene Nanoparticles Perturb Lipid Membranes.

    PubMed

    Rossi, Giulia; Barnoud, Jonathan; Monticelli, Luca

    2014-01-01

    Polystyrene is abundant in marine debris. Like most synthetic polymers, it degrades very slowly, producing smaller and smaller particles easily ingested by wildlife. The presence of plastic microscopic particles in fish and marine wildlife is massive and well documented, but its impact on cellular activity is not understood. Biological activity generally requires interaction with biological membranes, but this is difficult to study at the molecular scale in vivo. Here we use coarse-grained molecular simulations to determine the effect of nanosized polystyrene (PS) particles on the properties of model biological membranes. We find that PS nanoparticles permeate easily into lipid membranes. Dissolved in the membrane core, PS chains alter membrane structure, significantly reduce molecular diffusion, and soften the membrane. Moreover, PS severely affects membrane lateral organization by stabilizing raft-like domains. Changes in membrane properties and lateral organization can severely affect the activity of membrane proteins and thereby cellular function.

  20. Supported microporous ceramic membranes

    DOEpatents

    Webster, Elizabeth; Anderson, Marc

    1993-01-01

    A method for permformation of microporous ceramic membranes onto a porous support includes placing a colloidal suspension of metal or metal oxide particles on one side of the porous support and exposing the other side of the porous support to a drying stream of gas or a reactive gas stream so that the particles are deposited on the drying side of the support as a gel. The gel so deposited can be sintered to form a supported ceramic membrane useful for ultrafiltration, reverse osmosis, or molecular sieving having mean pore sizes less than 100 Angstroms.

  1. Supported microporous ceramic membranes

    DOEpatents

    Webster, E.; Anderson, M.

    1993-12-14

    A method for the formation of microporous ceramic membranes onto a porous support includes placing a colloidal suspension of metal or metal oxide particles on one side of the porous support and exposing the other side of the porous support to a drying stream of gas or a reactive gas stream so that the particles are deposited on the drying side of the support as a gel. The gel so deposited can be sintered to form a supported ceramic membrane useful for ultrafiltration, reverse osmosis, or molecular sieving having mean pore sizes less than 100 Angstroms. 4 figures.

  2. Battery utilizing ceramic membranes

    DOEpatents

    Yahnke, M.S.; Shlomo, G.; Anderson, M.A.

    1994-08-30

    A thin film battery is disclosed based on the use of ceramic membrane technology. The battery includes a pair of conductive collectors on which the materials for the anode and the cathode may be spin coated. The separator is formed of a porous metal oxide ceramic membrane impregnated with electrolyte so that electrical separation is maintained while ion mobility is also maintained. The entire battery can be made less than 10 microns thick while generating a potential in the 1 volt range. 2 figs.

  3. Spine-shaped gold protrusions improve the adherence and electrical coupling of neurons with the surface of micro-electronic devices

    PubMed Central

    Hai, Aviad; Dormann, Ada; Shappir, Joseph; Yitzchaik, Shlomo; Bartic, Carmen; Borghs, Gustaaf; Langedijk, J. P. M.; Spira, Micha E.

    2009-01-01

    Interfacing neurons with micro- and nano-electronic devices has been a subject of intense study over the last decade. One of the major problems in assembling efficient neuro-electronic hybrid systems is the weak electrical coupling between the components. This is mainly attributed to the fundamental property of living cells to form and maintain an extracellular cleft between the plasma membrane and any substrate to which they adhere. This cleft shunts the current generated by propagating action potentials and thus reduces the signal-to-noise ratio. Reducing the cleft thickness, and thereby increasing the seal resistance formed between the neurons and the sensing surface, is thus a challenge and could improve the electrical coupling coefficient. Using electron microscopic analysis and field potential recordings, we examined here the use of gold micro-structures that mimic dendritic spines in their shape and dimensions to improve the adhesion and electrical coupling between neurons and micro-electronic devices. We found that neurons cultured on a gold-spine matrix, functionalized by a cysteine-terminated peptide with a number of RGD repeats, readily engulf the spines, forming tight apposition. The recorded field potentials of cultured Aplysia neurons are significantly larger using gold-spine electrodes in comparison with flat electrodes. PMID:19474080

  4. Spine-shaped gold protrusions improve the adherence and electrical coupling of neurons with the surface of micro-electronic devices.

    PubMed

    Hai, Aviad; Dormann, Ada; Shappir, Joseph; Yitzchaik, Shlomo; Bartic, Carmen; Borghs, Gustaaf; Langedijk, J P M; Spira, Micha E

    2009-12-01

    Interfacing neurons with micro- and nano-electronic devices has been a subject of intense study over the last decade. One of the major problems in assembling efficient neuro-electronic hybrid systems is the weak electrical coupling between the components. This is mainly attributed to the fundamental property of living cells to form and maintain an extracellular cleft between the plasma membrane and any substrate to which they adhere. This cleft shunts the current generated by propagating action potentials and thus reduces the signal-to-noise ratio. Reducing the cleft thickness, and thereby increasing the seal resistance formed between the neurons and the sensing surface, is thus a challenge and could improve the electrical coupling coefficient. Using electron microscopic analysis and field potential recordings, we examined here the use of gold micro-structures that mimic dendritic spines in their shape and dimensions to improve the adhesion and electrical coupling between neurons and micro-electronic devices. We found that neurons cultured on a gold-spine matrix, functionalized by a cysteine-terminated peptide with a number of RGD repeats, readily engulf the spines, forming tight apposition. The recorded field potentials of cultured Aplysia neurons are significantly larger using gold-spine electrodes in comparison with flat electrodes.

  5. TOM1L1 drives membrane delivery of MT1-MMP to promote ERBB2-induced breast cancer cell invasion

    PubMed Central

    Chevalier, Clément; Collin, Guillaume; Descamps, Simon; Touaitahuata, Heiani; Simon, Valérie; Reymond, Nicolas; Fernandez, Laurent; Milhiet, Pierre-Emmanuel; Georget, Virginie; Urbach, Serge; Lasorsa, Laurence; Orsetti, Béatrice; Boissière-Michot, Florence; Lopez-Crapez, Evelyne; Theillet, Charles; Roche, Serge; Benistant, Christine

    2016-01-01

    ERBB2 overexpression in human breast cancer leads to invasive carcinoma but the mechanism is not clearly understood. Here we report that TOM1L1 is co-amplified with ERBB2 and defines a subgroup of HER2+/ER+ tumours with early metastatic relapse. TOM1L1 encodes a GAT domain-containing trafficking protein and is a SRC substrate that negatively regulates tyrosine kinase signalling. We demonstrate that TOM1L1 upregulation enhances the invasiveness of ERBB2-transformed cells. This pro-tumoural function does not involve SRC, but implicates membrane-bound membrane-type 1 MMP (MT1-MMP)-dependent activation of invadopodia, membrane protrusions specialized in extracellular matrix degradation. Mechanistically, ERBB2 elicits the indirect phosphorylation of TOM1L1 on Ser321. The phosphorylation event promotes GAT-dependent association of TOM1L1 with the sorting protein TOLLIP and trafficking of the metalloprotease MT1-MMP from endocytic compartments to invadopodia for tumour cell invasion. Collectively, these results show that TOM1L1 is an important element of an ERBB2-driven proteolytic invasive programme and that TOM1L1 amplification potentially enhances the metastatic progression of ERBB2-positive breast cancers. PMID:26899482

  6. TOM1L1 drives membrane delivery of MT1-MMP to promote ERBB2-induced breast cancer cell invasion.

    PubMed

    Chevalier, Clément; Collin, Guillaume; Descamps, Simon; Touaitahuata, Heiani; Simon, Valérie; Reymond, Nicolas; Fernandez, Laurent; Milhiet, Pierre-Emmanuel; Georget, Virginie; Urbach, Serge; Lasorsa, Laurence; Orsetti, Béatrice; Boissière-Michot, Florence; Lopez-Crapez, Evelyne; Theillet, Charles; Roche, Serge; Benistant, Christine

    2016-01-01

    ERBB2 overexpression in human breast cancer leads to invasive carcinoma but the mechanism is not clearly understood. Here we report that TOM1L1 is co-amplified with ERBB2 and defines a subgroup of HER2(+)/ER(+) tumours with early metastatic relapse. TOM1L1 encodes a GAT domain-containing trafficking protein and is a SRC substrate that negatively regulates tyrosine kinase signalling. We demonstrate that TOM1L1 upregulation enhances the invasiveness of ERBB2-transformed cells. This pro-tumoural function does not involve SRC, but implicates membrane-bound membrane-type 1 MMP (MT1-MMP)-dependent activation of invadopodia, membrane protrusions specialized in extracellular matrix degradation. Mechanistically, ERBB2 elicits the indirect phosphorylation of TOM1L1 on Ser321. The phosphorylation event promotes GAT-dependent association of TOM1L1 with the sorting protein TOLLIP and trafficking of the metalloprotease MT1-MMP from endocytic compartments to invadopodia for tumour cell invasion. Collectively, these results show that TOM1L1 is an important element of an ERBB2-driven proteolytic invasive programme and that TOM1L1 amplification potentially enhances the metastatic progression of ERBB2-positive breast cancers. PMID:26899482

  7. Characterization of a zeolite membrane for catalytic membrane reactor application

    SciTech Connect

    Giroir-Fendler, A.; Peureux, J.; Mozzanega, H.; Dalmon, J.A.

    1996-12-31

    This paper describes the morphological and transport properties of a composite zeolite (silicalite) - alumina membrane. Some advantages obtained in combining the membrane with a conventional fixed-bed catalyst are also reported.

  8. The interactions of peripheral membrane proteins with biological membranes

    DOE PAGES

    Johs, Alexander; Whited, A. M.

    2015-01-01

    The interactions of peripheral proteins with membrane surfaces are critical to many biological processes, including signaling, recognition, membrane trafficking, cell division and cell structure. On a molecular level, peripheral membrane proteins can modulate lipid composition, membrane dynamics and protein-protein interactions. Biochemical and biophysical studies have shown that these interactions are in fact highly complex, dominated by several different types of interactions, and have an interdependent effect on both the protein and membrane. Here we examine three major mechanisms underlying the interactions between peripheral membrane proteins and membranes: electrostatic interactions, hydrophobic interactions, and fatty acid modification of proteins. While experimental approachesmore » continue to provide critical insights into specific interaction mechanisms, emerging bioinformatics resources and tools contribute to a systems-level picture of protein-lipid interactions. Through these recent advances, we begin to understand the pivotal role of protein-lipid interactions underlying complex biological functions at membrane interfaces.« less

  9. The interactions of peripheral membrane proteins with biological membranes.

    PubMed

    Whited, A M; Johs, A

    2015-11-01

    The interactions of peripheral proteins with membrane surfaces are critical to many biological processes, including signaling, recognition, membrane trafficking, cell division and cell structure. On a molecular level, peripheral membrane proteins can modulate lipid composition, membrane dynamics and protein-protein interactions. Biochemical and biophysical studies have shown that these interactions are in fact highly complex, dominated by several different types of interactions, and have an interdependent effect on both the protein and membrane. Here we examine three major mechanisms underlying the interactions between peripheral membrane proteins and membranes: electrostatic interactions, hydrophobic interactions, and fatty acid modification of proteins. While experimental approaches continue to provide critical insights into specific interaction mechanisms, emerging bioinformatics resources and tools contribute to a systems-level picture of protein-lipid interactions. Through these recent advances, we begin to understand the pivotal role of protein-lipid interactions underlying complex biological functions at membrane interfaces.

  10. The interactions of peripheral membrane proteins with biological membranes

    SciTech Connect

    Johs, Alexander; Whited, A. M.

    2015-01-01

    The interactions of peripheral proteins with membrane surfaces are critical to many biological processes, including signaling, recognition, membrane trafficking, cell division and cell structure. On a molecular level, peripheral membrane proteins can modulate lipid composition, membrane dynamics and protein-protein interactions. Biochemical and biophysical studies have shown that these interactions are in fact highly complex, dominated by several different types of interactions, and have an interdependent effect on both the protein and membrane. Here we examine three major mechanisms underlying the interactions between peripheral membrane proteins and membranes: electrostatic interactions, hydrophobic interactions, and fatty acid modification of proteins. While experimental approaches continue to provide critical insights into specific interaction mechanisms, emerging bioinformatics resources and tools contribute to a systems-level picture of protein-lipid interactions. Through these recent advances, we begin to understand the pivotal role of protein-lipid interactions underlying complex biological functions at membrane interfaces.

  11. Enzyme catalytic membrane based on a hybrid mesoporous membrane.

    PubMed

    Fu, Wensheng; Yamaguchi, Akira; Kaneda, Hideaki; Teramae, Norio

    2008-02-21

    Immobilization of glucose oxidase (GOD) within a hybrid mesoporous membrane with 12 nm pore diameter was successfully achieved, resulting in catalytically high efficiency during flow of a glucose solution across the membrane. PMID:18253526

  12. Synthetic membranes and membrane processes with counterparts in biological systems

    NASA Astrophysics Data System (ADS)

    Matson, Stephen L.

    1996-02-01

    Conventional synthetic membranes, fashioned for the most part from rather unremarkable polymeric materials, are essentially passive structures that achieve various industrial and biomedical separations through simple and selective membrane permeation processes. Indeed, simplicity of membrane material, structure, and function has long been perceived as a virtue of membranes relative to other separation processes with which they compete. The passive membrane separation processes -- exemplified by micro- and ultrafiltration, dialysis, reverse osmosis, and gas permeation -- differ from one another primarily in terms of membrane morphology or structure (e.g., porous, gel-type, and nonporous) and the permeant transport mechanism and driving force (e.g., diffusion, convection, and 'solution/diffusion'). The passive membrane separation processes have in common the fact that interaction between permeant and membrane material is typically weak and physicochemical in nature; indeed, it is frequently an objective of membrane materials design to minimize interaction between permeant and membrane polymer, since such strategies can minimize membrane fouling. As a consequence, conventional membrane processes often provide only modest separation factors or permselectivities; that is, they are more useful in performing 'group separations' (i.e., the separation of different classes of material) than they are in fractionating species within a given class. It has long been recognized within the community of membrane technologists that biological membrane structures and their components are extraordinarily sophisticated and powerful as compared to their synthetic counterparts. Moreover, biomembranes and related biological systems have been 'designed' according to a very different paradigm -- one that frequently maximizes and capitalizes on extraordinarily strong and biochemically specific interactions between components of the membrane and species interacting with them. Thus, in recent

  13. Sequential Activities of Phosphoinositide 3-Kinase, PKB/Akt, and Rab7 during Macropinosome Formation in Dictyostelium

    PubMed Central

    Rupper, Adam; Lee, Kyung; Knecht, David; Cardelli, James

    2001-01-01

    Macropinocytosis plays an important role in the internalization of antigens by dendritic cells and is the route of entry for many bacterial pathogens; however, little is known about the molecular mechanisms that regulate the formation or maturation of macropinosomes. Like dendritic cells, Dictyostelium amoebae are active in macropinocytosis, and various proteins have been identified that contribute to this process. As described here, microscopic analysis of null mutants have revealed that the class I phosphoinositide 3-kinases, PIK1 and PIK2, and the downstream effector protein kinase B (PKB/Akt) are important in regulating completion of macropinocytosis. Although actin-rich membrane protrusions form in these cell lines, they recede without forming macropinosomes. Imaging of cells expressing green fluorescent protein (GFP) fused to the pleckstrin homology domain (PH) of PKB (GFP-PHPKB) indicates that D3 phosphoinositides are enriched in the forming macropinocytic cup and remain associated with newly formed macropinosomes for <1 minute. A fusion protein, consisting of GFP fused to an F-actin binding domain, overlaps with GFP-PHPKB in the timing of association with forming macropinosomes. Although macropinocytosis is reduced in cells expressing dominant negative Rab7, microscopic imaging studies reveal that GFP-Rab7 associates only with formed macropinosomes at approximately the time that F-actin and D3 phosphoinositide levels decrease. These results support a model in which F-actin modulating proteins and vesicle trafficking proteins coordinately regulate the formation and maturation of macropinosomes. PMID:11553719

  14. The neuronal proteins CIPP, Cypin and IRSp53 form a tripartite complex mediated by PDZ and SH3 domains.

    PubMed

    Barilari, Manuela; Dente, Luciana

    2010-10-01

    Here we report the dissection of a tripartite complex formed by CIPP (channel-interacting PDZ protein), IRSp53 (insulin receptor tyrosine kinase substrate protein) and Cypin (cytosolic PSD-95 interactor) in cultured cells. The three proteins are expressed in similar neuronal districts, where CIPP binds to different membrane channels and receptors, IRSp53 regulates the morphogenesis of actin-rich dendritic spines, and Cypin promotes dendrite branching and patterning by binding to tubulin heterodimers. We observed that the interaction among the three proteins is mediated by small binding domains: CIPP works as a bridge, linking the carboxy-termini of IRSp53 and Cypin with its PDZ domains; IRSp53 connects Cypin, through an unusual SH3-mediated association, which can be impaired by substituting two crucial positively charged residues of Cypin. The observation that the three engineered proteins co-localize in the cytoplasm, and at the tip of induced neurites in neuronal cells, raises the interesting possibility that they work together in the formation of neuronal protrusions.

  15. The cancer-testis antigens SPANX-A/C/D and CTAG2 promote breast cancer invasion

    PubMed Central

    Prechtl, Amanda M.; Dang, Tuyen T.; Whitehurst, Angelique W.; Pearson, Gray W.

    2016-01-01

    Genes that are normally biased towards expression in the testis are often induced in tumor cells. These gametogenic genes, known as cancer-testis antigens (CTAs), have been extenstively investigated as targets for immunotherapy. However, despite their frequent detection, the degree to which CTAs support neoplastic invasion is poorly understood. Here, we find that the CTA genes SPANX-A/C/D and CTAG2 are coordinately induced in breast cancer cells and regulate distinct features of invasive behavior. Our functional analysis revealed that CTAG2 interacts with Pericentrin at the centrosome and is necessary for directional migration. Conversely, SPANX-A/C/D interacts with Lamin A/C at the inner nuclear membrane and is required for the formation of actin-rich cellular protrusions that reorganize the extracellular matrix. Importantly, SPANX-A/C/D was required for breast cancer cells to spontaneously metastasize to the lung, demonstrating that CTA reactivation can be critical for invasion dependent phenotypes in vivo. Moreover, elevated SPANX-A/C/D expression in breast cancer patient tumors correlated with poor outcome. Together, our results suggest that distinct CTAs promote tumor progression by regulating complementary cellular functions that are integrated together to induce invasive behavior. PMID:26895102

  16. The cancer-testis antigens SPANX-A/C/D and CTAG2 promote breast cancer invasion.

    PubMed

    Maine, Erin A; Westcott, Jill M; Prechtl, Amanda M; Dang, Tuyen T; Whitehurst, Angelique W; Pearson, Gray W

    2016-03-22

    Genes that are normally biased towards expression in the testis are often induced in tumor cells. These gametogenic genes, known as cancer-testis antigens (CTAs), have been extenstively investigated as targets for immunotherapy. However, despite their frequent detection, the degree to which CTAs support neoplastic invasion is poorly understood. Here, we find that the CTA genes SPANX-A/C/D and CTAG2 are coordinately induced in breast cancer cells and regulate distinct features of invasive behavior. Our functional analysis revealed that CTAG2 interacts with Pericentrin at the centrosome and is necessary for directional migration. Conversely, SPANX-A/C/D interacts with Lamin A/C at the inner nuclear membrane and is required for the formation of actin-rich cellular protrusions that reorganize the extracellular matrix. Importantly, SPANX-A/C/D was required for breast cancer cells to spontaneously metastasize to the lung, demonstrating that CTA reactivation can be critical for invasion dependent phenotypes in vivo. Moreover, elevated SPANX-A/C/D expression in breast cancer patient tumors correlated with poor outcome. Together, our results suggest that distinct CTAs promote tumor progression by regulating complementary cellular functions that are integrated together to induce invasive behavior.

  17. Supervillin Reorganizes the Actin Cytoskeleton and Increases Invadopodial Efficiency

    PubMed Central

    Crowley, Jessica L.; Smith, Tara C.; Fang, Zhiyou; Takizawa, Norio

    2009-01-01

    Tumor cells use actin-rich protrusions called invadopodia to degrade extracellular matrix (ECM) and invade tissues; related structures, termed podosomes, are sites of dynamic ECM interaction. We show here that supervillin (SV), a peripheral membrane protein that binds F-actin and myosin II, reorganizes the actin cytoskeleton and potentiates invadopodial function. Overexpressed SV induces redistribution of lamellipodial cortactin and lamellipodin/RAPH1/PREL1 away from the cell periphery to internal sites and concomitantly increases the numbers of F-actin punctae. Most punctae are highly dynamic and colocalize with the podosome/invadopodial proteins, cortactin, Tks5, and cdc42. Cortactin binds SV sequences in vitro and contributes to the formation of enhanced green fluorescent protein (EGFP)-SV induced punctae. SV localizes to the cores of Src-generated podosomes in COS-7 cells and with invadopodia in MDA-MB-231 cells. EGFP-SV overexpression increases average numbers of ECM holes per cell; RNA interference-mediated knockdown of SV decreases these numbers. Although SV knockdown alone has no effect, simultaneous down-regulation of SV and the closely related protein gelsolin reduces invasion through ECM. Together, our results show that SV is a component of podosomes and invadopodia and that SV plays a role in invadopodial function, perhaps as a mediator of cortactin localization, activation state, and/or dynamics of metalloproteinases at the ventral cell surface. PMID:19109420

  18. Endophilin A1 regulates dendritic spine morphogenesis and stability through interaction with p140Cap

    PubMed Central

    Yang, Yanrui; Wei, Mengping; Xiong, Ying; Du, Xiangyang; Zhu, Shaoxia; Yang, Lin; Zhang, Chen; Liu, Jia-Jia

    2015-01-01

    Dendritic spines are actin-rich membrane protrusions that are the major sites of excitatory synaptic input in the mammalian brain, and their morphological plasticity provides structural basis for learning and memory. Here we report that endophilin A1, with a well-established role in clathrin-mediated synaptic vesicle endocytosis at the presynaptic terminal, also localizes to dendritic spines and is required for spine morphogenesis, synapse formation and synaptic function. We identify p140Cap, a regulator of cytoskeleton reorganization, as a downstream effector of endophilin A1 and demonstrate that disruption of their interaction impairs spine formation and maturation. Moreover, we demonstrate that knockdown of endophilin A1 or p140Cap impairs spine stabilization and synaptic function. We further show that endophilin A1 regulates the distribution of p140Cap and its downstream effector, the F-actin-binding protein cortactin as well as F-actin enrichment in dendritic spines. Together, these results reveal a novel function of postsynaptic endophilin A1 in spine morphogenesis, stabilization and synaptic function through the regulation of p140Cap. PMID:25771685

  19. Rapid Remodeling of Invadosomes by Gi-coupled Receptors: DISSECTING THE ROLE OF Rho GTPases.

    PubMed

    Kedziora, Katarzyna M; Leyton-Puig, Daniela; Argenzio, Elisabetta; Boumeester, Anja J; van Butselaar, Bram; Yin, Taofei; Wu, Yi I; van Leeuwen, Frank N; Innocenti, Metello; Jalink, Kees; Moolenaar, Wouter H

    2016-02-26

    Invadosomes are actin-rich membrane protrusions that degrade the extracellular matrix to drive tumor cell invasion. Key players in invadosome formation are c-Src and Rho family GTPases. Invadosomes can reassemble into circular rosette-like superstructures, but the underlying signaling mechanisms remain obscure. Here we show that Src-induced invadosomes in human melanoma cells (A375M and MDA-MB-435) undergo rapid remodeling into dynamic extracellular matrix-degrading rosettes by distinct G protein-coupled receptor agonists, notably lysophosphatidic acid (LPA; acting through the LPA1 receptor) and endothelin. Agonist-induced rosette formation is blocked by pertussis toxin, dependent on PI3K activity and accompanied by localized production of phosphatidylinositol 3,4,5-trisphosphate, whereas MAPK and Ca(2+) signaling are dispensable. Using FRET-based biosensors, we show that LPA and endothelin transiently activate Cdc42 through Gi, concurrent with a biphasic decrease in Rac activity and differential effects on RhoA. Cdc42 activity is essential for rosette formation, whereas G12/13-mediated RhoA-ROCK signaling suppresses the remodeling process. Our results reveal a Gi-mediated Cdc42 signaling axis by which G protein-coupled receptors trigger invadosome remodeling, the degree of which is dictated by the Cdc42-RhoA activity balance. PMID:26740622

  20. Cholesterol-Dependent Membrane Fusion Induced by the gp41 Membrane-Proximal External Region–Transmembrane Domain Connection Suggests a Mechanism for Broad HIV-1 Neutralization

    PubMed Central

    Apellániz, Beatriz; Rujas, Edurne; Carravilla, Pablo; Requejo-Isidro, José; Huarte, Nerea

    2014-01-01

    ABSTRACT The HIV-1 glycoprotein 41 promotes fusion of the viral membrane with that of the target cell. Structural, biochemical, and biophysical studies suggest that its membrane-proximal external region (MPER) may interact with the HIV-1 membrane and induce its disruption and/or deformation during the process. However, the high cholesterol content of the envelope (ca. 40 to 50 mol%) imparts high rigidity, thereby acting against lipid bilayer restructuring. Here, based on the outcome of vesicle stability assays, all-atom molecular dynamics simulations, and atomic force microscopy observations, we propose that the conserved sequence connecting the MPER with the N-terminal residues of the transmembrane domain (TMD) is involved in HIV-1 fusion. This junction would function by inducing phospholipid protrusion and acyl-chain splay in the cholesterol-enriched rigid envelope. Supporting the functional relevance of such a mechanism, membrane fusion was inhibited by the broadly neutralizing 4E10 antibody but not by a nonneutralizing variant with the CDR-H3 loop deleted. We conclude that the MPER-TMD junction embodies an envelope-disrupting C-terminal fusion peptide that can be targeted by broadly neutralizing antibodies. IMPORTANCE Fusion of the cholesterol-enriched viral envelope with the cell membrane marks the beginning of the infectious HIV-1 replicative cycle. Consequently, the Env glycoprotein-mediated fusion function constitutes an important clinical target for inhibitors and preventive vaccines. Antibodies 4E10 and 10E8 bind to one Env vulnerability site located at the gp41 membrane-proximal external region (MPER)–transmembrane domain (TMD) junction and block infection. These antibodies display broad viral neutralization, which underscores the conservation and functionality of the MPER-TMD region. In this work, we combined biochemical assays with molecular dynamics simulations and microscopy observations to characterize the unprecedented fusogenic activity of the

  1. Membrane Transfer Phenomena (MTP)

    NASA Technical Reports Server (NTRS)

    Mason, Larry

    1996-01-01

    Progress has been made in several areas of the definition, design, and development of the Membrane Transport Apparatus (MTA) instrument and associated sensors and systems. Progress is also reported in the development of software modules for instrument control, experimental image and data acquisition, and data analysis.

  2. Membranes in motion

    PubMed Central

    van Meer, Gerrit

    2010-01-01

    The Keystone Symposium on the Molecular Basis for Biological Membrane Organization and Dynamics held in January this year offered new insights into the molecular machines at work in cells. Topics included the machinery responsible for the dynamic shape of organelles, the budding and fusion of vesicular carriers, and the intricate sorting systems that ensure the correct delivery of cellular components. PMID:20395954

  3. Hydrogen-Selective Membrane

    DOEpatents

    Collins, John P.; Way, J. Douglas

    1995-09-19

    A hydrogen-selective membrane comprises a tubular porous ceramic support having a palladium metal layer deposited on an inside surface of the ceramic support. The thickness of the palladium layer is greater than about 10 .mu.m but typically less than about 20 .mu.m. The hydrogen permeation rate of the membrane is greater than about 1.0 moles/m.sup.2.s at a temperature of greater than about 500.degree. C. and a transmembrane pressure difference of about 1,500 kPa. Moreover, the hydrogen-to-nitrogen selectivity is greater than about 600 at a temperature of greater than about 500.degree. C. and a transmembrane pressure of about 700 kPa. Hydrogen can be separated from a mixture of gases using the membrane. The method may include the step of heating the mixture of gases to a temperature of greater than about 400.degree. C. and less than about 1000.degree. C. before the step of flowing the mixture of gases past the membrane. The mixture of gases may include ammonia. The ammonia typically is decomposed to provide nitrogen and hydrogen using a catalyst such as nickel. The catalyst may be placed inside the tubular ceramic support. The mixture of gases may be supplied by an industrial process such as the mixture of exhaust gases from the IGCC process.

  4. Hydrogen-selective membrane

    DOEpatents

    Collins, J.P.; Way, J.D.

    1995-09-19

    A hydrogen-selective membrane comprises a tubular porous ceramic support having a palladium metal layer deposited on an inside surface of the ceramic support. The thickness of the palladium layer is greater than about 10 {micro}m but typically less than about 20 {micro}m. The hydrogen permeation rate of the membrane is greater than about 1.0 moles/m{sup 2}s at a temperature of greater than about 500 C and a transmembrane pressure difference of about 1,500 kPa. Moreover, the hydrogen-to-nitrogen selectivity is greater than about 600 at a temperature of greater than about 500 C and a transmembrane pressure of about 700 kPa. Hydrogen can be separated from a mixture of gases using the membrane. The method may include the step of heating the mixture of gases to a temperature of greater than about 400 C and less than about 1000 C before the step of flowing the mixture of gases past the membrane. The mixture of gases may include ammonia. The ammonia typically is decomposed to provide nitrogen and hydrogen using a catalyst such as nickel. The catalyst may be placed inside the tubular ceramic support. The mixture of gases may be supplied by an industrial process such as the mixture of exhaust gases from the IGCC process. 9 figs.

  5. Hydrogen-selective membrane

    DOEpatents

    Collins, J.P.; Way, J.D.

    1997-07-29

    A hydrogen-selective membrane comprises a tubular porous ceramic support having a palladium metal layer deposited on an inside surface of the ceramic support. The thickness of the palladium layer is greater than about 10 {micro}m but typically less than about 20 {micro}m. The hydrogen permeation rate of the membrane is greater than about 1.0 moles/m{sup 2} s at a temperature of greater than about 500 C and a transmembrane pressure difference of about 1,500 kPa. Moreover, the hydrogen-to-nitrogen selectivity is greater than about 600 at a temperature of greater than about 500 C and a transmembrane pressure of about 700 kPa. Hydrogen can be separated from a mixture of gases using the membrane. The method may include the step of heating the mixture of gases to a temperature of greater than about 400 C and less than about 1000 C before the step of flowing the mixture of gases past the membrane. The mixture of gases may include ammonia. The ammonia typically is decomposed to provide nitrogen and hydrogen using a catalyst such as nickel. The catalyst may be placed inside the tubular ceramic support. The mixture of gases may be supplied by an industrial process such as the mixture of exhaust gases from the IGCC process. 9 figs.

  6. Hydrogen-selective membrane

    DOEpatents

    Collins, John P.; Way, J. Douglas

    1997-01-01

    A hydrogen-selective membrane comprises a tubular porous ceramic support having a palladium metal layer deposited on an inside surface of the ceramic support. The thickness of the palladium layer is greater than about 10 .mu.m but typically less than about 20 .mu.m. The hydrogen permeation rate of the membrane is greater than about 1.0 moles/m.sup.2. s at a temperature of greater than about 500.degree. C. and a transmembrane pressure difference of about 1,500 kPa. Moreover, the hydrogen-to-nitrogen selectivity is greater than about 600 at a temperature of greater than about 500.degree. C. and a transmembrane pressure of about 700 kPa. Hydrogen can be separated from a mixture of gases using the membrane. The method may include the step of heating the mixture of gases to a temperature of greater than about 400.degree. C. and less than about 1000.degree. C. before the step of flowing the mixture of gases past the membrane. The mixture of gases may include ammonia. The ammonia typically is decomposed to provide nitrogen and hydrogen using a catalyst such as nickel. The catalyst may be placed inside the tubular ceramic support. The mixture of gases may be supplied by an industrial process such as the mixture of exhaust gases from the IGCC process.

  7. Improved ion exchange membrane

    NASA Technical Reports Server (NTRS)

    Rembaum, A.; Yen, S. P. S.; Klein, E.

    1975-01-01

    Membrane, made from commercially-available hollow fibers, is used in reverse osmosis, or dialysis. Fiber has skin layers which pass only small molecules. Macromolecules cannot penetrate skin. Fibers can also be used to remove other undesirable anions, such as phosphate, sulfate, carbonate, and uranium in form of uranium-sulfate complex.

  8. Effective rigidity of membranes

    NASA Astrophysics Data System (ADS)

    Peliti, L.

    1986-12-01

    The role of thermal fluctuations of shape (undulations) in reducing the effective rigidity of membranes is reviewed. The consequences of this effect on vesicle size distribution and on the structure of microemulsions, as well as on other physical phenomena, are sketched.

  9. Premature rupture of membranes

    MedlinePlus

    ... the 37th week of pregnancy, it is called preterm premature rupture of membranes (PPROM). The earlier your water breaks, the more ... babies born early). If there is not a preterm unit where you deliver, you and your baby will be moved to a hospital that has one.

  10. Fluctuating nematic elastomer membranes.

    PubMed

    Xing, Xiangjun; Mukhopadhyay, Ranjan; Lubensky, T C; Radzihovsky, Leo

    2003-08-01

    We study the flat phase of nematic elastomer membranes with rotational symmetry spontaneously broken by an in-plane nematic order. Such a state is characterized by a vanishing elastic modulus for simple shear and soft transverse phonons. At harmonic level, the in-plane orientational (nematic) order is stable to thermal fluctuations that lead to short-range in-plane translational (phonon) correlations. To treat thermal fluctuations and relevant elastic nonlinearities, we introduce two generalizations of two-dimensional membranes in a three-dimensional space to arbitrary D-dimensional membranes embedded in a d-dimensional space and analyze their anomalous elasticities in an expansion about D=4. We find a stable fixed point that controls long-scale properties of nematic elastomer membranes. It is characterized by singular in-plane elastic moduli that vanish as a power law eta(lambda)=4-D of a relevant inverse length scale (e.g., wave vector) and a finite bending rigidity. Our predictions are asymptotically exact near four dimensions. PMID:14524954

  11. Fluctuating nematic elastomer membranes

    NASA Astrophysics Data System (ADS)

    Xing, Xiangjun; Mukhopadhyay, Ranjan; Lubensky, T. C.; Radzihovsky, Leo

    2003-08-01

    We study the flat phase of nematic elastomer membranes with rotational symmetry spontaneously broken by an in-plane nematic order. Such a state is characterized by a vanishing elastic modulus for simple shear and soft transverse phonons. At harmonic level, the in-plane orientational (nematic) order is stable to thermal fluctuations that lead to short-range in-plane translational (phonon) correlations. To treat thermal fluctuations and relevant elastic nonlinearities, we introduce two generalizations of two-dimensional membranes in a three-dimensional space to arbitrary D-dimensional membranes embedded in a d-dimensional space and analyze their anomalous elasticities in an expansion about D=4. We find a stable fixed point that controls long-scale properties of nematic elastomer membranes. It is characterized by singular in-plane elastic moduli that vanish as a power law ηλ=4-D of a relevant inverse length scale (e.g., wave vector) and a finite bending rigidity. Our predictions are asymptotically exact near four dimensions.

  12. Membrane protein secretases.

    PubMed Central

    Hooper, N M; Karran, E H; Turner, A J

    1997-01-01

    A diverse range of membrane proteins of Type 1 or Type II topology also occur as a circulating, soluble form. These soluble forms are often derived from the membrane form by proteolysis by a group of enzymes referred to collectively as 'secretases' or 'sheddases'. The cleavage generally occurs close to the extracellular face of the membrane, releasing physiologically active protein. This secretion process also provides a mechanism for down-regulating the protein at the cell surface. Examples of such post-translational proteolysis are seen in the Alzheimer's amyloid precursor protein, the vasoregulatory enzyme angiotensin converting enzyme, transforming growth factor-alpha, the tumour necrosis factor ligand and receptor superfamilies, certain cytokine receptors, and others. Since the proteins concerned are involved in pathophysiological processes such as neurodegeneration, apoptosis, oncogenesis and inflammation, the secretases could provide novel therapeutic targets. Recent characterization of these individual secretases has revealed common features, particularly sensitivity to certain metalloprotease inhibitors and upregulation of activity by phorbol esters. It is therefore likely that a closely related family of metallosecretases controls the surface expression of multiple integral membrane proteins. Current knowledge of the various secretases are compared in this Review, and strategies for cell-free assays of such proteases are outlined as a prelude to their ultimate purification and cloning. PMID:9020855

  13. Membrane humidity control investigation

    NASA Technical Reports Server (NTRS)

    Elam, J.; Ruder, J.; Strumpf, H.

    1974-01-01

    The basic performance data on a hollow fiber membrane unit that removes water from a breathing gas loop by diffusion is presented. Using available permeability data for cellulose acetate, a preliminary design was made of a dehumidifier unit that would meet the problem statement.

  14. Composite oxygen transport membrane

    DOEpatents

    Christie, Gervase Maxwell; Lane, Jonathan A.

    2014-08-05

    A method of producing a composite oxygen ion membrane and a composite oxygen ion membrane in which a porous fuel oxidation layer and a dense separation layer and optionally, a porous surface exchange layer are formed on a porous support from mixtures of (Ln.sub.1-xA.sub.x).sub.wCr.sub.1-yB.sub.yO.sub.3-.delta. and a doped zirconia. In the porous fuel oxidation layer and the optional porous surface exchange layer, A is Calcium and in the dense separation layer A is not Calcium and, preferably is Strontium. Preferred materials are (La.sub.0.8Ca.sub.0.2).sub.0.95Cr.sub.0.5Mn.sub.0.5O.sub.3-.delta. for the porous fuel oxidation and optional porous surface exchange layers and (La.sub.0.8Sr.sub.0.2).sub.0.95Cr.sub.0.5Fe.sub.0.5O.sub.3-.delta. for the dense separation layer. The use of such materials allows the membrane to sintered in air and without the use of pore formers to reduce membrane manufacturing costs. The use of materials, as described herein, for forming the porous layers have application for forming any type of porous structure, such as a catalyst support.

  15. Dynamics of multicomponent lipid membranes

    NASA Astrophysics Data System (ADS)

    Camley, Brian Andrew

    We present theoretical and computational descriptions of the dynamics of multicomponent lipid bilayer membranes. These systems are both model systems for "lipid rafts" in cell membranes and interesting physical examples of quasi-two-dimensional fluids. Our chief tool is a continuum simulation that uses a phase field to track the composition of the membrane, and solves the hydrodynamic equations exactly using the appropriate Green's function (Oseen tensor) for the membrane. We apply this simulation to describe the diffusion of domains in phase-separated membranes, the dynamics of domain flickering, and the process of phase separation in lipid membranes. We then derive an analytical theory to describe domain flickering that is consistent with our simulation results, and use this to analyze experimental measurements of membrane domains. Through this method, we measure the membrane viscosity solely from fluorescence microscopy measurements. We study phase separation in quasi-two-dimensional membranes in depth with both simulations and scaling theory, and classify the different scaling regimes and morphologies, which differ from pure two-dimensional fluids. Our results may explain previous inconsistent measurements of the dynamical scaling exponent for phase separation in membranes. We also extend our theory beyond the simplest model, including the possibility that the membrane will be viscoelastic, as well as considering the inertia of the membrane and the fluid surrounding the membrane.

  16. HYDROGEN SEPARATION MEMBRANES

    SciTech Connect

    Donald P. McCollor; John P. Kay

    1999-08-01

    A likely membrane for future testing of high-temperature hydrogen separation from a gasification product stream was targeted as an inorganic analog of a dense-metal membrane, where the hydrogen would dissolve into and diffuse through the membrane structure. An amorphous membrane such as zinc sulfide appeared to be promising. Previously, ZnS film coating tests had been performed using an electron-beam vacuum coating instrument, with zinc films successfully applied to glass substrates. The coatings appeared relatively stable in air and in a simple simulated gasification atmosphere at elevated temperature. Because the electron-beam coating instrument suffered irreparable breakdown, several alternative methods were tested in an effort to produce a nitrogen-impermeable, hydrogen-permeable membrane on porous sintered steel substrates. None of the preparation methods proved successful in sealing the porous substrate against nitrogen gas. To provide a nitrogen-impermeable ZnS material to test for hydrogen permeability, two ZnS infrared sample windows were purchased. These relatively thick ''membranes'' did not show measurable permeation of hydrogen, either due to lack of absorption or a negligible permeation rate due to their thickness. To determine if hydrogen was indeed adsorbed, thermogravimetric and differential thermal analyses tests were performed on samples of ZnS powder. A significant uptake of hydrogen gas occurred, corresponding to a maximum of 1 mole H{sub 2} per 1 mole ZnS at a temperature of 175 C. The hydrogen remained in the material at ambient temperature in a hydrogen atmosphere, but approximately 50% would be removed in argon. Reheating in a hydrogen atmosphere resulted in no additional hydrogen uptake. Differential scanning calorimetry indicated that the hydrogen uptake was probably due to the formation of a zinc-sulfur-hydrogen species resulting in the formation of hydrogen sulfide. The zinc sulfide was found to be unstable above approximately 200 C

  17. Alternative energy efficient membrane bioreactor using reciprocating submerged membrane.

    PubMed

    Ho, J; Smith, S; Roh, H K

    2014-01-01

    A novel membrane bioreactor (MBR) pilot system, using membrane reciprocation instead of air scouring, was operated at constant high flux and daily fluctuating flux to demonstrate its application under peak and diurnal flow conditions. Low and stable transmembrane pressure was achieved at 40 l/m(2)/h (LMH) by use of repetitive membrane reciprocation. The results reveal that the inertial forces acting on the membrane fibers effectively propel foulants from the membrane surface. Reciprocation of the hollow fiber membrane is beneficial for the constant removal of solids that may build up on the membrane surface and inside the membrane bundle. The membrane reciprocation in the reciprocating MBR pilot consumed less energy than coarse air scouring used in conventional MBR systems. Specific energy consumption for the membrane reciprocation was 0.072 kWh/m(3) permeate produced at 40 LMH flux, which is 75% less than for a conventional air scouring system as reported in literature without consideration of energy consumption for biological aeration (0.29 kWh/m(3)). The daily fluctuating flux test confirmed that the membrane reciprocation is effective to handle fluctuating flux up to 50 LMH. The pilot-scale reciprocating MBR system successfully demonstrated that fouling can be controlled via 0.43 Hz membrane reciprocation with 44 mm or higher amplitude.

  18. Water Fluxes in Polymeric Membranes for Desalination via Membrane Distillation

    NASA Astrophysics Data System (ADS)

    Mannella, G. A.; Brucato, V.; La Carrubba, V.

    2010-06-01

    Membrane distillation is an emerging technique for seawater desalination. Hydrophobic polymeric membranes are used to separate the solute-free water vapour from the hot solution. Vapour fluxes of commercial polymeric membranes were measured in various conditions, i.e. natural and forced convection and vacuum. Vapour fluxes were also predicted with models and compared with experimentals. Higher fluxes were recorded in vacuum conditions.

  19. Selective permeability of PVA membranes. I - Radiation-crosslinked membranes

    NASA Technical Reports Server (NTRS)

    Katz, M. G.; Wydeven, T., Jr.

    1981-01-01

    The water and salt transport properties of ionizing radiation crosslinked poly(vinyl alcohol) (PVA) membranes were investigated. The studied membranes showed high permeabilities and low selectivities for both water and salt. The results were found to be in accord with a modified solution-diffusion model for transport across the membranes, in which pressure-dependent permeability coefficients are employed.

  20. Selective Permeability of PVA Membranes. I: Radiation-Crosslinked Membranes

    NASA Technical Reports Server (NTRS)

    Katz, Moshe G.; Wydeven, Theodore, Jr.

    1981-01-01

    The water and salt transport properties of ionizing radiation crosslinked poly(vinyl alcohol) (PVA) membranes were investigated. The studied membranes showed high permeabilities and low selectivities for both water and salt. The results were found to be in accord with a modified solution-diffusion model for transport across the membranes, in which pressure-dependent permeability coefficients are employed.

  1. Process of treating cellulosic membrane and alkaline with membrane separator

    NASA Technical Reports Server (NTRS)

    Hoyt, H. E.; Pfluger, H. L. (Inventor)

    1970-01-01

    The improvement of water-soluble cellulose ether membranes for use as separators in concentrated alkaline battery cells is discussed. The process of contacting membranes with an aqueous alkali solution of concentration less than that of the alkali solution to be used in the battery but above that at which the membrane is soluble is described.

  2. Alternative energy efficient membrane bioreactor using reciprocating submerged membrane.

    PubMed

    Ho, J; Smith, S; Roh, H K

    2014-01-01

    A novel membrane bioreactor (MBR) pilot system, using membrane reciprocation instead of air scouring, was operated at constant high flux and daily fluctuating flux to demonstrate its application under peak and diurnal flow conditions. Low and stable transmembrane pressure was achieved at 40 l/m(2)/h (LMH) by use of repetitive membrane reciprocation. The results reveal that the inertial forces acting on the membrane fibers effectively propel foulants from the membrane surface. Reciprocation of the hollow fiber membrane is beneficial for the constant removal of solids that may build up on the membrane surface and inside the membrane bundle. The membrane reciprocation in the reciprocating MBR pilot consumed less energy than coarse air scouring used in conventional MBR systems. Specific energy consumption for the membrane reciprocation was 0.072 kWh/m(3) permeate produced at 40 LMH flux, which is 75% less than for a conventional air scouring system as reported in literature without consideration of energy consumption for biological aeration (0.29 kWh/m(3)). The daily fluctuating flux test confirmed that the membrane reciprocation is effective to handle fluctuating flux up to 50 LMH. The pilot-scale reciprocating MBR system successfully demonstrated that fouling can be controlled via 0.43 Hz membrane reciprocation with 44 mm or higher amplitude. PMID:25521136

  3. OXYGEN TRANSPORT CERAMIC MEMBRANES

    SciTech Connect

    Dr. Sukumar Bandopadhyay; Dr. Nagendra Nagabhushana

    2001-12-01

    Conversion of natural gas to liquid fuels and chemicals is a major goal for the Nation as it enters the 21st Century. Technically robust and economically viable processes are needed to capture the value of the vast reserves of natural gas on Alaska's North Slope, and wean the Nation from dependence on foreign petroleum sources. Technologies that are emerging to fulfill this need are all based syngas as an intermediate. Syngas (a mixture of hydrogen and carbon monoxide) is a fundamental building block from which chemicals and fuels can be derived. Lower cost syngas translates directly into more cost-competitive fuels and chemicals. The currently practiced commercial technology for making syngas is either steam methane reforming (SMR) or a two-step process involving cryogenic oxygen separation followed by natural gas partial oxidation (POX). These high-energy, capital-intensive processes do not always produce syngas at a cost that makes its derivatives competitive with current petroleum-based fuels and chemicals. This project has the following 6 main tasks: Task 1--Design, fabricate and evaluate ceramic to metal seals based on graded ceramic powder/metal braze joints. Task 2--Evaluate the effect of defect configuration on ceramic membrane conductivity and long term chemical and structural stability. Task 3--Determine materials mechanical properties under conditions of high temperatures and reactive atmospheres. Task 4--Evaluate phase stability and thermal expansion of candidate perovskite membranes and develop techniques to support these materials on porous metal structures. Task 5--Assess the microstructure of membrane materials to evaluate the effects of vacancy-impurity association, defect clusters, and vacancy-dopant association on the membrane performance and stability. Task 6--Measure kinetics of oxygen uptake and transport in ceramic membrane materials under commercially relevant conditions using isotope labeling techniques.

  4. Membrane Cells for Brine Electrolysis.

    ERIC Educational Resources Information Center

    Tingle, M.

    1982-01-01

    Membrane cells were developed as alternatives to mercury and diaphragm cells for the electrolysis of brine. Compares the three types of cells, focusing on the advantages and disadvantages of membrane cells. (JN)

  5. Ion Channels in Nerve Membranes

    ERIC Educational Resources Information Center

    Ehrenstein, Gerald

    1976-01-01

    Discusses research that indicates that nerve membranes, which play a key role in the conduction of impulses, are traversed by protein channels with ion pathways opened and closed by the membrane electric field. (Author/MLH)

  6. Fuel cell with ionization membrane

    NASA Technical Reports Server (NTRS)

    Hartley, Frank T. (Inventor)

    2007-01-01

    A fuel cell is disclosed comprising an ionization membrane having at least one area through which gas is passed, and which ionizes the gas passing therethrough, and a cathode for receiving the ions generated by the ionization membrane. The ionization membrane may include one or more openings in the membrane with electrodes that are located closer than a mean free path of molecules within the gas to be ionized. Methods of manufacture are also provided.

  7. Active membrane phased array radar

    NASA Technical Reports Server (NTRS)

    Moussessian, Alina; Del Castillo, Linda; Huang, John; Sadowy, Greg; Hoffman, James; Smith, Phil; Hatake, Toshiro; Derksen, Chuck; Lopez, Bernardo; Caro, Ed

    2005-01-01

    We have developed the first membrane-based active phased array in L-band (1.26GHz). The array uses membrane compatible Transmit/Receive (T/R) modules (membrane T/R) for each antenna element. We use phase shifters within each T/R module for electronic beam steering. We will discuss the T/R module design and integration with the membrane, We will also present transmit and receive beam-steering results for the array.

  8. Structural Analysis of the Roles of Influenza A Virus Membrane-Associated Proteins in Assembly and Morphology

    PubMed Central

    Chlanda, Petr; Schraidt, Oliver; Kummer, Susann; Riches, James; Oberwinkler, Heike; Prinz, Simone; Kräusslich, Hans-Georg

    2015-01-01

    ABSTRACT The assembly of influenza A virus at the plasma membrane of infected cells leads to release of enveloped virions that are typically round in tissue culture-adapted strains but filamentous in strains isolated from patients. The viral proteins hemagglutinin (HA), neuraminidase (NA), matrix protein 1 (M1), and M2 ion channel all contribute to virus assembly. When expressed individually or in combination in cells, they can all, under certain conditions, mediate release of membrane-enveloped particles, but their relative roles in virus assembly, release, and morphology remain unclear. To investigate these roles, we produced membrane-enveloped particles by plasmid-derived expression of combinations of HA, NA, and M proteins (M1 and M2) or by infection with influenza A virus. We monitored particle release, particle morphology, and plasma membrane morphology by using biochemical methods, electron microscopy, electron tomography, and cryo-electron tomography. Our data suggest that HA, NA, or HANA (HA plus NA) expression leads to particle release through nonspecific induction of membrane curvature. In contrast, coexpression with the M proteins clusters the glycoproteins into filamentous membrane protrusions, which can be released as particles by formation of a constricted neck at the base. HA and NA are preferentially distributed to differently curved membranes within these particles. Both the budding intermediates and the released particles are morphologically similar to those produced during infection with influenza A virus. Together, our data provide new insights into influenza virus assembly and show that the M segment together with either of the glycoproteins is the minimal requirement to assemble and release membrane-enveloped particles that are truly virus-like. IMPORTANCE Influenza A virus is a major respiratory pathogen. It assembles membrane-enveloped virus particles whose shapes vary from spherical to filamentous. Here we examine the roles of individual

  9. Pretreatment and Membrane Hydrophilic Modification to Reduce Membrane Fouling

    PubMed Central

    Sun, Wen; Liu, Junxia; Chu, Huaqiang; Dong, Bingzhi

    2013-01-01

    The application of low pressure membranes (microfiltration/ultrafiltration) has undergone accelerated development for drinking water production. However, the major obstacle encountered in its popularization is membrane fouling caused by natural organic matter (NOM). This paper firstly summarizes the two factors causing the organic membrane fouling, including molecular weight (MW) and hydrophilicity/hydrophobicity of NOM, and then presents a brief introduction of the methods which can prevent membrane fouling such as pretreatment of the feed water (e.g., coagulation, adsorption, and pre-oxidation) and membrane hydrophilic modification (e.g., plasma modification, irradiation grafting modification, surface coating modification, blend modification, etc.). Perspectives of further research are also discussed. PMID:24956947

  10. Variable tilt on lipid membranes

    PubMed Central

    Rangamani, P.; Steigmann, D. J.

    2014-01-01

    A continuum theory for lipid membranes is developed that accounts for mechanical interactions between lipid tilt and membrane shape. For planar membranes, a linear version of the theory is used to predict tilt variations similar to those observed in experiments and molecular dynamics simulations. PMID:25484606

  11. An Electrochromic Bipolar Membrane Diode.

    PubMed

    Malti, Abdellah; Gabrielsson, Erik O; Crispin, Xavier; Berggren, Magnus

    2015-07-01

    Conducting polymers with bipolar membranes (a complementary stack of selective membranes) may be used to rectify current. Integrating a bipolar membrane into a polymer electrochromic display obviates the need for an addressing backplane while increasing the device's bistability. Such devices can be made from solution-processable materials.

  12. Entropic Tension in Crowded Membranes

    PubMed Central

    Lindén, Martin; Sens, Pierre; Phillips, Rob

    2012-01-01

    Unlike their model membrane counterparts, biological membranes are richly decorated with a heterogeneous assembly of membrane proteins. These proteins are so tightly packed that their excluded area interactions can alter the free energy landscape controlling the conformational transitions suffered by such proteins. For membrane channels, this effect can alter the critical membrane tension at which they undergo a transition from a closed to an open state, and therefore influence protein function in vivo. Despite their obvious importance, crowding phenomena in membranes are much less well studied than in the cytoplasm. Using statistical mechanics results for hard disk liquids, we show that crowding induces an entropic tension in the membrane, which influences transitions that alter the projected area and circumference of a membrane protein. As a specific case study in this effect, we consider the impact of crowding on the gating properties of bacterial mechanosensitive membrane channels, which are thought to confer osmoprotection when these cells are subjected to osmotic shock. We find that crowding can alter the gating energies by more than in physiological conditions, a substantial fraction of the total gating energies in some cases. Given the ubiquity of membrane crowding, the nonspecific nature of excluded volume interactions, and the fact that the function of many membrane proteins involve significant conformational changes, this specific case study highlights a general aspect in the function of membrane proteins. PMID:22438801

  13. Membrane tension and peripheral protein density mediate membrane shape transitions

    NASA Astrophysics Data System (ADS)

    Shi, Zheng; Baumgart, Tobias

    2015-01-01

    Endocytosis is a ubiquitous eukaryotic membrane budding, vesiculation and internalization process fulfilling numerous roles including compensation of membrane area increase after bursts of exocytosis. The mechanism of the coupling between these two processes to enable homeostasis is not well understood. Recently, an ultrafast endocytosis (UFE) pathway was revealed with a speed significantly exceeding classical clathrin-mediated endocytosis (CME). Membrane tension reduction is a potential mechanism by which endocytosis can be rapidly activated at remote sites. Here, we provide experimental evidence for a mechanism whereby membrane tension reduction initiates membrane budding and tubulation mediated by endocytic proteins, such as endophilin A1. We find that shape instabilities occur at well-defined membrane tensions and surface densities of endophilin. From our data, we obtain a membrane shape stability diagram that shows remarkable consistency with a quantitative model. This model applies to all laterally diffusive curvature-coupling proteins and therefore a wide range of endocytic proteins.

  14. Elastic deformation and area per lipid of membranes: atomistic view from solid-state deuterium NMR spectroscopy.

    PubMed

    Kinnun, Jacob J; Mallikarjunaiah, K J; Petrache, Horia I; Brown, Michael F

    2015-01-01

    This article reviews the application of solid-state ²H nuclear magnetic resonance (NMR) spectroscopy for investigating the deformation of lipid bilayers at the atomistic level. For liquid-crystalline membranes, the average structure is manifested by the segmental order parameters (SCD) of the lipids. Solid-state ²H NMR yields observables directly related to the stress field of the lipid bilayer. The extent to which lipid bilayers are deformed by osmotic pressure is integral to how lipid-protein interactions affect membrane functions. Calculations of the average area per lipid and related structural properties are pertinent to bilayer remodeling and molecular dynamics (MD) simulations of membranes. To establish structural quantities, such as area per lipid and volumetric bilayer thickness, a mean-torque analysis of ²H NMR order parameters is applied. Osmotic stress is introduced by adding polymer solutions or by gravimetric dehydration, which are thermodynamically equivalent. Solid-state NMR studies of lipids under osmotic stress probe membrane interactions involving collective bilayer undulations, order-director fluctuations, and lipid molecular protrusions. Removal of water yields a reduction of the mean area per lipid, with a corresponding increase in volumetric bilayer thickness, by up to 20% in the liquid-crystalline state. Hydrophobic mismatch can shift protein states involving mechanosensation, transport, and molecular recognition by G-protein-coupled receptors. Measurements of the order parameters versus osmotic pressure yield the elastic area compressibility modulus and the corresponding bilayer thickness at an atomistic level. Solid-state ²H NMR thus reveals how membrane deformation can affect protein conformational changes within the stress field of the lipid bilayer.

  15. Membrane separation of hydrocarbons

    DOEpatents

    Chang, Y. Alice; Kulkarni, Sudhir S.; Funk, Edward W.

    1986-01-01

    Mixtures of heavy oils and light hydrocarbons may be separated by passing the mixture through a polymeric membrane. The membrane which is utilized to effect the separation comprises a polymer which is capable of maintaining its integrity in the presence of hydrocarbon compounds and which has been modified by being subjected to the action of a sulfonating agent. Sulfonating agents which may be employed will include fuming sulfuric acid, chlorosulfonic acid, sulfur trioxide, etc., the surface or bulk modified polymer will contain a degree of sulfonation ranging from about 15 to about 50%. The separation process is effected at temperatures ranging from about ambient to about 100.degree. C. and pressures ranging from about 50 to about 1000 psig.

  16. Oxygen Transport Ceramic Membranes

    SciTech Connect

    S. Bandopadhyay; N. Nagabhushana; T. Nithyanantham; X.-D Zhou; Y-W. Sin; H.U. Anderson; Alan Jacobson; C.A. Mims

    2005-02-01

    The present quarterly report describes some of the investigations on the structural properties of dense OTM bars provided by Praxair and studies on newer composition of Ti doped LSF. Thermogravimetric analysis (TGA) was carried out on La{sub 0.2}Sr{sub 0.8}Fe{sub 0.55}Ti{sub 0.45}O{sub 3-{delta}} to investigate oxygen deficiency ({delta}) of the sample. The TGA was performed in a controlled atmosphere using oxygen, argon, carbon monoxide and carbon dioxide with adjustable gas flow rates. In this experiment, the weight loss and gain of La{sub 0.2}Sr{sub 0.8}Fe{sub 0.55}Ti{sub 0.45}O{sub 3-{delta}} was directly measured by TGA. The weight change of the sample was evaluated at between 600 and 1250 C in air or 1000 C as a function of oxygen partial pressure. The oxygen deficiencies calculated from TGA data as a function of oxygen activity and temperature will be estimated and compared with that from neutron diffraction measurement in air. The LSFT and LSFT/CGO membranes were fabricated from the powder obtained from Praxair Specialty Ceramics. The sintered membranes were subjected to microstructure analysis and hardness analysis. The LSFT membrane is composed of fine grains with two kinds of grain morphology. The grain size distribution was characterized using image analysis. In LSFT/CGO membrane a lot of grain pullout was observed from the less dense, porous phase. The hardness of the LSFT and dual phase membranes were studied at various loads. The hardness values obtained from the cross section of the membranes were also compared to that of the values obtained from the surface. An electrochemical cell has been designed and built for measurements of the Seebeck coefficient as a function of temperature and pressure. Measurements on La{sub 0.2}Sr{sub 0.8}Fe{sub 0.55}Ti{sub 0.45}O{sub 3-{delta}} as a function of temperature an oxygen partial pressure are reported. Further analysis of the dilatometry data obtained previously is presented. A series of isotope transients

  17. The First Cell Membranes

    NASA Technical Reports Server (NTRS)

    Deamer, David; Dworkin, Jason P.; Sandford, Scott A.; Bernstein, Max P.; Allamandola, Louis J.

    2004-01-01

    Organic compounds are synthesized in the interstellar medium and can be delivered to planetary surfaces such as the early Earth, where they mix with endogenous organic mixtures. Some of these compounds are amphiphilic, having polar and non-polar groups on the same molecule. Amphiphilic compounds spontaneously self-assembly into more complex structures such as bimolecular layers, which in turn form closed membranous vesicles. The first forms of cellular life required self-assembled membranes that were likely to be available on the prebiotic Earth. Laboratory simulations show that such vesicles readily encapsulate functional macromolecules, including nucleic acids and polymerases. A goal of future investigations is to fabricate artificial cells as models of the origin of life.

  18. OXYGEN TRANSPORT CERAMIC MEMBRANES

    SciTech Connect

    Dr. Sukumar Bandopadhyay; Dr. Nagendra Nagabhushana

    2003-01-01

    In the present quarter, experiments are presented on ceramic/metal interactions of Zirconia/Ni-B-Si system and with a thin Ti coating deposited on zirconia surface. Processing of perovskites of LSC, LSF and LSCF composition for evaluation of mechanical properties as a function of environment are begun. The studies are to be in parallel with LSFCO composition to characterize the segregation of cations and slow crack growth in environmental conditions. La{sub 1-x}Sr{sub x}FeO{sub 3-d} has also been characterized for paramagnetic ordering at room temperature and the evolution of magnetic moments as a function of temperature are investigated. Investigation on the thermodynamic properties of the membrane materials are continued to develop a complete model for the membrane transport.

  19. Recycling of used perfluorosulfonic acid membranes

    DOEpatents

    Grot, Stephen; Grot, Walther

    2007-08-14

    A method for recovering and recycling catalyst coated fuel cell membranes includes dissolving the used membranes in water and solvent, heating the dissolved membranes under pressure and separating the components. Active membranes are produced from the recycled materials.

  20. NASA In-step: Permeable Membrane Experiment

    NASA Technical Reports Server (NTRS)

    1992-01-01

    Viewgraphs on the Permeable Membrane Experiment are presented. An experiment overview is given. The Membrane Phase Separation Experiment, Membrane Diffusion Interference Experiment, and Membrane Wetting Experiment are described. Finally, summary and conclusions are discussed.

  1. Membrane Stability Testing

    SciTech Connect

    Hobbs, D.T.

    1997-09-30

    The Electrosynthesis Co. Inc. (ESC) was contracted by the Westinghouse Savannah River Company to investigate the long term performance and durability of cell components (anode, membrane, cathode) in an electrochemical caustic recovery process using a simulated SRC liquid waste as anolyte solution. This report details the results of two long-term studies conducted using an ICI FM01 flow cell. This cell is designed and has previously been demonstrated to scale up directly into the commercial scale ICI FM21 cell.

  2. HSPES membrane electrode assembly

    NASA Technical Reports Server (NTRS)

    Kindler, Andrew (Inventor); Yen, Shiao-Ping (Inventor)

    2000-01-01

    An improved fuel cell electrode, as well as fuel cells and membrane electrode assemblies that include such an electrode, in which the electrode includes a backing layer having a sintered layer thereon, and a non-sintered free-catalyst layer. The invention also features a method of forming the electrode by sintering a backing material with a catalyst material and then applying a free-catalyst layer.

  3. OXYGEN TRANSPORT CERAMIC MEMBRANES

    SciTech Connect

    Dr. Sukumar Bandopadhyay; Dr. Nagendra Nagabhushana

    2000-07-01

    This is the fourth quarterly report on a new study to develop a ceramic membrane/metal joint. The first experiments using the La-Sr-Fe-O ceramic are reported. Some of the analysis performed on the samples obtained are commented upon. A set of experiments to characterize the mechanical strength and thermal fatigue properties of the joints has been designed and begun. Finite element models of joints used to model residual stresses are described.

  4. Membrane Transport Phenomena (MTP)

    NASA Technical Reports Server (NTRS)

    Mason, Larry W.

    1997-01-01

    The third semi-annual period of the MTP project has been involved with performing experiments using the Membrane Transport Apparatus (MTA), development of analysis techniques for the experiment results, analytical modeling of the osmotic transport phenomena, and completion of a DC-9 microgravity flight to test candidate fluid cell geometries. Preparations were also made for the MTP Science Concept Review (SCR), held on 13 June 1997 at Lockheed Martin Astronautics in Denver. These activities are detailed in the report.

  5. Characterization and expression of plasma and tonoplast membrane aquaporins in primed seed of Brassica napus during germination under stress conditions.

    PubMed

    Gao, Y P; Young, L; Bonham-Smith, P; Gusta, L V

    1999-07-01

    Two aquaporin genes were isolated from a cDNA library of canola (Brassica napus L.). The first aquaporin, BnPIP1 of 1094 bp, encoding a putative polypeptide of 287 amino acids with a predicted molecular mass of 30.4 kDa and a pI of 7.8, belongs to the family of plasma membrane intrinsic protein (PIPs) aquaporins. The B. napus aquaporin showed 85-94% identity to the Arabidopsis thaliana PIPs. ABA priming of seed induced high levels of BnPIP1 transcript which remained after subsequent re-drying of the seed. The second aquaporin, Bny-TIP2 of 1020 bp, encoded a putative polypeptide of 253 amino acids with a predicted molecular mass of 25.8 kDa and a pI of 5.8. Bngamma-TIP2 showed 83-90% identity to gamma-TIP genes from a variety of plant species. Bngamma-TIP2 was expressed only when radicle protrusion occurred in either untreated or primed seeds. Seeds primed with PEG or ABA germinated earlier and showed a higher final percentage of germination than unprimed seed, particularly under salt and osmotic stresses at low temperature. Transcripts of both BnPIP1 and Bngamma-TIP2 genes were present earlier during germination of primed seeds than non-primed seed. From these results, we conclude that BnPIP1 is related to the water transportation required for enzymatic metabolism of storage nutrients at the early stages of canola seed germination whereas Bngamma-TIP2 expression is related to cell growth associated with radicle protrusion. Priming induced the expression of BnPIP1 but had no effect on Bngamma-TIP2.

  6. Water vapor diffusion membrane development

    NASA Technical Reports Server (NTRS)

    Tan, M. K.

    1976-01-01

    A total of 18 different membranes were procured, characterized, and tested in a modified bench-scale vapor diffusion water reclamation unit. Four membranes were selected for further studies involving membrane fouling. Emphasis was placed on the problem of flux decline due to membrane fouling. This is discussed in greater details under "Summary and Discussion on Membrane Fouling Studies" presented in pages 47-51. The system was also investigated for low temperature application on wash-water where the permeated water is not recovered but vented into space vacuum.

  7. Premature rupture of membranes.

    PubMed Central

    Poma, P. A.

    1996-01-01

    The management of patients with premature rupture of membranes has changed markedly in the past several years. The basis for this is a combination of a better understanding of newborn physiology, improved neonatal care, refinements in antibiotic therapy, and the widespread use of maternal and fetal monitoring. The best outcome for both mother and infant undoubtedly reflects data based on a combination of factors, among which are gestational age survival, evidence of fetal distress, presence or absence of labor and sepsis, and of course, the cervical condition as it is related to labor-readiness. An important recent advance is the recognition that an active observation management program is associated with less morbidity and mortality than the classic management course of delivery within 12 hours of membrane rupture. The fact that preterm premature rupture of membranes tends to recur in subsequent pregnancies offers an opportunity for prevention. Moreover, advances in perinatal and neonatal care will continue to improve the outcomes of these women and their children. PMID:8583489

  8. Virus separation using membranes.

    PubMed

    Grein, Tanja A; Michalsky, Ronald; Czermak, Peter

    2014-01-01

    Industrial manufacturing of cell culture-derived viruses or virus-like particles for gene therapy or vaccine production are complex multistep processes. In addition to the bioreactor, such processes require a multitude of downstream unit operations for product separation, concentration, or purification. Similarly, before a biopharmaceutical product can enter the market, removal or inactivation of potential viral contamination has to be demonstrated. Given the complexity of biological solutions and the high standards on composition and purity of biopharmaceuticals, downstream processing is the bottleneck in many biotechnological production trains. Membrane-based filtration can be an economically attractive and efficient technology for virus separation. Viral clearance, for instance, of up to seven orders of magnitude has been reported for state of the art polymeric membranes under best conditions.This chapter summarizes the fundamentals of virus ultrafiltration, diafiltration, or purification with adsorptive membranes. In lieu of an impractical universally applicable protocol for virus filtration, application of these principles is demonstrated with two examples. The chapter provides detailed methods for production, concentration, purification, and removal of a rod-shaped baculovirus (Autographa californica M nucleopolyhedrovirus, about 40 × 300 nm in size, a potential vector for gene therapy, and an industrially important protein expression system) or a spherical parvovirus (minute virus of mice, 22-26 nm in size, a model virus for virus clearance validation studies).

  9. Novel Catalytic Membrane Reactors

    SciTech Connect

    Stuart Nemser, PhD

    2010-10-01

    There are many industrial catalytic organic reversible reactions with amines or alcohols that have water as one of the products. Many of these reactions are homogeneously catalyzed. In all cases removal of water facilitates the reaction and produces more of the desired chemical product. By shifting the reaction to right we produce more chemical product with little or no additional capital investment. Many of these reactions can also relate to bioprocesses. Given the large number of water-organic compound separations achievable and the ability of the Compact Membrane Systems, Inc. (CMS) perfluoro membranes to withstand these harsh operating conditions, this is an ideal demonstration system for the water-of-reaction removal using a membrane reactor. Enhanced reaction synthesis is consistent with the DOE objective to lower the energy intensity of U.S. industry 25% by 2017 in accord with the Energy Policy Act of 2005 and to improve the United States manufacturing competitiveness. The objective of this program is to develop the platform technology for enhancing homogeneous catalytic chemical syntheses.

  10. Lipid exchange between membranes.

    PubMed Central

    Jähnig, F

    1984-01-01

    The exchange of lipid molecules between vesicle bilayers in water and a monolayer forming at the water surface was investigated theoretically within the framework of thermodynamics. The total number of exchanged molecules was found to depend on the bilayer curvature as expressed by the vesicle radius and on the boundary condition for exchange, i.e., whether during exchange the radius or the packing density of the vesicles remains constant. The boundary condition is determined by the rate of flip-flop within the bilayer relative to the rate of exchange between bi- and monolayer. If flip-flop is fast, exchange is independent of the vesicle radius; if flip-flop is slow, exchange increases with the vesicle radius. Available experimental results agree with the detailed form of this dependence. When the theory was extended to exchange between two bilayers of different curvature, the direction of exchange was also determined by the curvatures and the boundary conditions for exchange. Due to the dependence of the boundary conditions on flip-flop and, consequently, on membrane fluidity, exchange between membranes may partially be regulated by membrane fluidity. PMID:6518251

  11. Viral membrane fusion

    SciTech Connect

    Harrison, Stephen C.

    2015-05-15

    Membrane fusion is an essential step when enveloped viruses enter cells. Lipid bilayer fusion requires catalysis to overcome a high kinetic barrier; viral fusion proteins are the agents that fulfill this catalytic function. Despite a variety of molecular architectures, these proteins facilitate fusion by essentially the same generic mechanism. Stimulated by a signal associated with arrival at the cell to be infected (e.g., receptor or co-receptor binding, proton binding in an endosome), they undergo a series of conformational changes. A hydrophobic segment (a “fusion loop” or “fusion peptide”) engages the target-cell membrane and collapse of the bridging intermediate thus formed draws the two membranes (virus and cell) together. We know of three structural classes for viral fusion proteins. Structures for both pre- and postfusion conformations of illustrate the beginning and end points of a process that can be probed by single-virion measurements of fusion kinetics. - Highlights: • Viral fusion proteins overcome the high energy barrier to lipid bilayer merger. • Different molecular structures but the same catalytic mechanism. • Review describes properties of three known fusion-protein structural classes. • Single-virion fusion experiments elucidate mechanism.

  12. Painted supported lipid membranes

    PubMed Central

    Florin, E.-L.; Gaub, H. E.

    1993-01-01

    We report herein measurements on a novel type of supported lipid films, which we call painted supported membranes (PSM). These membranes are formed in a self-assembly process on alkylated gold films from an organic solution. The formation process was investigated with surface plasmon resonance microscopy. The optical and electrical properties of the films were determined for various types of lipids and as a function of temperature by means of cyclic voltammetry and potential relaxation after charge injection. We could show that these films exhibit an extraordinarily high specific resistivity which, depending on the lipid, may be as high as 109 ohm/cm2. We could also show that due to this low conductivity, an electrical polarization across the PSM relaxes with characteristic time constants of up to 20 min. The electrical properties together with their high mechanical stability and accessibility to surface sensitive techniques make these films well suitable model membranes for optical and electrical investigations. Examples for such applications are given in the subsequent article by Seifert et al. ImagesFIGURE 3FIGURE 4 PMID:19431873

  13. Superhydrophobicity on transparent fluorinated ethylene propylene films with nano-protrusion morphology by Ar + O{sub 2} plasma etching: Study of the degradation in hydrophobicity after exposure to the environment

    SciTech Connect

    Gupta, Nitant; Kavya, M. V.; Singh, Yogesh R. G.; Jyothi, J.; Barshilia, Harish C.

    2013-10-28

    Fluorinated ethylene propylene (FEP) films were made superhydrophobic by Ar + O{sub 2} plasma etching process. Field emission scanning electron microscopy and atomic force microscopy studies of the plasma-treated FEP samples detected the presence of uniformly distributed nano-protrusions exhibiting a low surface roughness necessary for maintaining the transparency of the samples. In fact, optical transmittance measurements showed an improvement in the transparency of FEP samples after plasma treatment. The X-ray photoelectron spectroscopic analysis showed the presence of –CF{sub x}–O–CF{sub x}– (x = 1, 2, or 3) linkages in both untreated and plasma-treated samples which explains the hydrophilic nature (contact angle below 90{sup ∘}) of the untreated sample. Fourier transform infrared spectroscopy showed no changes in the bulk properties of the plasma-treated samples. Moreover, exposure to the environment caused the surfaces to lose their superhydrophobic property in an indefinite amount of time. This has been further studied through a water immersion experiment and explained through the wetting state transition from Cassie state to Wenzel state.

  14. Microtechnologies for membrane protein studies

    PubMed Central

    Suzuki, Hiroaki

    2008-01-01

    Despite the rapid and enormous progress in biotechnologies, the biochemical analysis of membrane proteins is still a difficult task. The presence of the large hydrophobic region buried in the lipid bilayer membrane (transmembrane domain) makes it difficult to analyze membrane proteins in standard assays developed for water-soluble proteins. To handle membrane proteins, the lipid bilayer membrane may be used as a platform to sustain their functionalities. Relatively slow progress in developing micro total analysis systems (μTAS) for membrane protein analysis directly reflects the difficulty of handling lipid membranes, which is a common problem in bulk measurement technologies. Nonetheless, researchers are continuing to develop efficient and sensitive analytical microsystems for the study of membrane proteins. Here, we review the latest developments, which enable detection of events caused by membrane proteins, such as ion channel current, membrane transport, and receptor/ligand interaction, by utilizing microfabricated structures. High-throughput and highly sensitive detection systems for membrane proteins are now becoming a realistic goal. Although most of these systems are still in the early stages of development, we believe this field will become one of the most important applications of μTAS for pharmaceutical and clinical screenings as well as for basic biochemical research. PMID:18335213

  15. How some proteins tubulate membranes

    NASA Astrophysics Data System (ADS)

    Bassereau, Patricia

    2009-03-01

    Endocytosis, exocytosis, membrane transport between intracellular compartments, virus or toxin entry or exit out of the cell, all imply to deform membrane. Membrane deformation mechanisms of cell membranes by proteins are currently actively studied. Giant vesicles (GUV) are interesting model membrane systems because they are composed of a very limited number of components compared to cellular membranes. The deformations induced by the interaction with a specific protein or any other additional components to the system, can then be directly monitored and the deformation mechanism eventually understood. In this talk, we will focus on different tubular structures induced by proteins. We will show that the B-subunits of Shiga toxin or Cholera Toxin, binding to their lipid receptors, Gb3 or GM1 respectively, incorporated in GUV membrane, induce negative membrane curvature and form tubular invaginations, in absence of any other cellular machinery. Tubular structures can also be obtained when molecular motors walking along microtubules exert a pulling force on the membrane of GUV. The helicoidal assembly of dynamin, a protein involved in vivo in membrane fission can also produce tubular structures. This assembly has been reconstituted around membrane nanotubes of controlled diameter; we will show that the initial tube diameter strongly influences dynamin polymerisation. In each case, a physical framework for understanding deformation mechanism will be presented

  16. Lipopolysaccharide Membrane Building and Simulation

    PubMed Central

    Jo, Sunhwan; Wu, Emilia L.; Stuhlsatz, Danielle; Klauda, Jeffery B.; Widmalm, Göran; Im, Wonpil

    2015-01-01

    Summary While membrane simulations are widely employed to study the structure and dynamics of various lipid bilayers and membrane proteins in the bilayers, simulations of lipopolysaccharides (LPS) in membrane environments have been limited due to its structural complexity, difficulties in building LPS-membrane systems, and lack of appropriate molecular force field. In this work, as a first step to extend CHARMM-GUI Membrane Builder to incorporate LPS molecules and to explore their structures and dynamics in membrane environments using molecular dynamics simulations, we describe step-by-step procedures to build LPS bilayer systems using CHARMM and the recently developed CHARMM carbohydrate and lipid force fields. Such procedures are illustrated by building various bilayers of Escherichia coli O6 LPS and their preliminary simulation results are given in terms of per-LPS area and density distributions of various components along the membrane normal. PMID:25753722

  17. Wrapping of nanoparticles by membranes.

    PubMed

    Bahrami, Amir H; Raatz, Michael; Agudo-Canalejo, Jaime; Michel, Raphael; Curtis, Emily M; Hall, Carol K; Gradzielski, Michael; Lipowsky, Reinhard; Weikl, Thomas R

    2014-06-01

    How nanoparticles interact with biomembranes is central for understanding their bioactivity. Biomembranes wrap around nanoparticles if the adhesive interaction between the nanoparticles and membranes is sufficiently strong to compensate for the cost of membrane bending. In this article, we review recent results from theory and simulations that provide new insights on the interplay of bending and adhesion energies during the wrapping of nanoparticles by membranes. These results indicate that the interplay of bending and adhesion during wrapping is strongly affected by the interaction range of the particle-membrane adhesion potential, by the shape of the nanoparticles, and by shape changes of membrane vesicles during wrapping. The interaction range of the particle-membrane adhesion potential is crucial both for the wrapping process of single nanoparticles and the cooperative wrapping of nanoparticles by membrane tubules.

  18. Solid-state membrane module

    DOEpatents

    Gordon, John Howard; Taylor, Dale M.

    2011-06-07

    Solid-state membrane modules comprising at least one membrane unit, where the membrane unit has a dense mixed conducting oxide layer, and at least one conduit or manifold wherein the conduit or manifold comprises a dense layer and at least one of a porous layer and a slotted layer contiguous with the dense layer. The solid-state membrane modules may be used to carry out a variety of processes including the separating of any ionizable component from a feedstream wherein such ionizable component is capable of being transported through a dense mixed conducting oxide layer of the membrane units making up the membrane modules. For ease of construction, the membrane units may be planar.

  19. CHARMM-GUI Membrane Builder Toward Realistic Biological Membrane Simulations

    PubMed Central

    Wu, Emilia L.; Cheng, Xi; Jo, Sunhwan; Rui, Huan; Song, Kevin C.; Dávila-Contreras, Eder M.; Qi, Yifei; Lee, Jumin; Monje-Galvan, Viviana; Venable, Richard M.; Klauda, Jeffery B.; Im, Wonpil

    2014-01-01

    CHARMM-GUI Membrane Builder, http://www.charmm-gui.org/input/membrane, is a web-based user interface designed to interactively build all-atom protein/membrane or membrane-only systems for molecular dynamics simulation through an automated optimized process. In this work, we describe the new features and major improvements in Membrane Builderthat allow users to robustly build realistic biological membrane systems, including (1) addition of new lipid types such as phosphoinositides, cardiolipin, sphingolipids, bacterial lipids, and ergosterol, yielding more than 180 lipid types, (2) enhanced building procedure for lipid packing around protein, (3) reliable algorithm to detect lipid tail penetration to ring structures and protein surface, (4) distance-based algorithm for faster initial ion displacement, (5) CHARMM inputs for P21 image transformation, and (6) NAMD equilibration and production inputs. The robustness of these new features is illustrated by building and simulating a membrane model of the polar and septal regions of E. coli membrane, which contains five lipid types: cardiolipin lipids with two types of acyl chains and phosphatidylethanolamine lipids with three types of acyl chains. It is our hope that CHARMM-GUI Membrane Builder becomes a useful tool for simulation studies to better understand the structure and dynamics of proteins and lipids in realistic biological membrane environments. PMID:25130509

  20. Membranes, methods of making membranes, and methods of separating gases using membranes

    DOEpatents

    Ho, W. S. Winston

    2012-10-02

    Membranes, methods of making membranes, and methods of separating gases using membranes are provided. The membranes can include at least one hydrophilic polymer, at least one cross-linking agent, at least one base, and at least one amino compound. The methods of separating gases using membranes can include contacting a gas stream containing at least one of CO.sub.2, H.sub.2S, and HCl with one side of a nonporous and at least one of CO.sub.2, H.sub.2S, and HCl selectively permeable membrane such that at least one of CO.sub.2, H.sub.2S, and HCl is selectively transported through the membrane.

  1. Membrane-mediated interactions measured using membrane domains.

    PubMed

    Semrau, Stefan; Idema, Timon; Schmidt, Thomas; Storm, Cornelis

    2009-06-17

    Cell membrane organization is the result of the collective effect of many driving forces. Several of these, such as electrostatic and van der Waals forces, have been identified and studied in detail. In this article, we investigate and quantify another force, the interaction between inclusions via deformations of the membrane shape. For electrically neutral systems, this interaction is the dominant organizing force. As a model system to study membrane-mediated interactions, we use phase-separated biomimetic vesicles that exhibit coexistence of liquid-ordered and liquid-disordered lipid domains. The membrane-mediated interactions between these domains lead to a rich variety of effects, including the creation of long-range order and the setting of a preferred domain size. Our findings also apply to the interaction of membrane protein patches, which induce similar membrane shape deformations and hence experience similar interactions.

  2. Chemical degradation mechanisms of membranes for alkaline membrane fuel cells

    SciTech Connect

    Choe, Yoong-Kee; Henson, Neil J.; Kim, Yu Seung

    2015-12-31

    Chemical degradation mechanisms of membranes for alkaline membrane fuel cells have been investigated using density functional theory (DFT). We have elucidated that the aryl-ether moiety of membranes is one of the weakest site against attack of hydroxide ions. The results of DFT calculations for hydroxide initiated aryl-ether cleavage indicated that the aryl-ether cleavage occurred prior to degradation of cationic functional group. Such a weak nature of the aryl-ether group arises from the electron deficiency of the aryl group as well as the low bond dissociation energy. The DFT results suggests that removal of the aryl-ether group in the membrane should enhance the stability of membranes under alkaline conditions. In fact, an ether fee poly(phenylene) membrane exhibits excellent stability against the attack from hydroxide ions.

  3. Oxygen Transport Ceramic Membranes

    SciTech Connect

    S. Bandopadhyay; T. Nithyanantham; X.-D Zhou; Y-W. Sin; H.U. Anderson; Alan Jacobson; C.A. Mims

    2005-08-01

    The present quarterly report describes some of the investigations on the structural properties of dense OTM bars provided by Praxair and studies on newer composition of Ti doped LSF. In the previous research, the reference point of oxygen occupancy was determined and verified. In the current research, the oxygen occupancy was investigated at 1200 C as a function of oxygen activity and compared with that at 1000 C. The cause of bumps at about 200 C was also investigated by using different heating and cooling rates during TGA. The fracture toughness of LSFT and dual phase membranes at room temperature is an important mechanical property. Vicker's indentation method was used to evaluate this toughness. Through this technique, a K{sub Ic} (Mode-I Fracture Toughness) value is attained by means of semi-empirical correlations between the indentation load and the length of the cracks emanating from the corresponding Vickers indentation impression. In the present investigation, crack propagation behavior was extensively analyzed in order to understand the strengthening mechanisms involved in the non-transforming La based ceramic composites. Cracks were generated using Vicker's indenter and used to identify and evaluate the toughening mechanisms involved. Preliminary results of an electron microscopy study of the origin of the slow kinetics on reduction of ferrites have been obtained. The slow kinetics appear to be related to a non-equilibrium reduction pathway that initially results in the formation of iron particles. At long times, equilibrium can be reestablished with recovery of the perovskite phase. Modeling of the isotopic transients on operating membranes (LSCrF-2828 at 900 C) and a ''frozen'' isotope profile have been analyzed in conjunction with a 1-D model to reveal the gradient in oxygen diffusivity through the membrane under conditions of high chemical gradients.

  4. Oxygen Transport Ceramic Membranes

    SciTech Connect

    S. Bandopadhyay; T. Nithyanantham; X.-D Zhou; Y-W. Sin; H.U. Anderson; Alan Jacobson; C.A. Mims

    2005-02-01

    The present quarterly report describes some of the investigations on the structural properties of dense OTM bars provided by Praxair and studies on newer composition of Ti doped LSF. The in situ electrical conductivity and Seebeck coefficient measurements were made on LSFT at 1000 and 1200 C over the oxygen activity range from air to 10{sup -15} atm. The electrical conductivity measurements exhibited a p to n type transition at an oxygen activity of 1 x 10{sup -10} at 1000 C and 1 x 10{sup -6} at 1200 C. Thermogravimetric studies were also carried out over the same oxygen activities and temperatures. Based on the results of these measurements, the chemical and mechanical stability range of LSFT were determined and defect structure was established. The studies on the fracture toughness of the LSFT and dual phase membranes exposed to air and N{sub 2} at 1000 C was done and the XRD and SEM analysis of the specimens were carried out to understand the structural and microstructural changes. The membranes that are exposed to high temperatures at an inert and a reactive atmosphere undergo many structural and chemical changes which affect the mechanical properties. A complete transformation of fracture behavior was observed in the N{sub 2} treated LSFT samples. Further results to investigate the origin of the slow kinetics on reduction of ferrites have been obtained. The slow kinetics appear to be related to a non-equilibrium reduction pathway that initially results in the formation of iron particles. At long times, equilibrium can be reestablished with recovery of the perovskite phase. Recent results on transient kinetic data are presented. The 2-D modeling of oxygen movement has been undertaken in order to fit isotope data. The model is used to study ''frozen'' profiles in patterned or composite membranes.

  5. Oxygen Transport Membranes

    SciTech Connect

    S. Bandopadhyay

    2008-08-30

    The focus of this research was to develop new membrane materials by synthesizing different compounds and determining their defect structures, crystallographic structures and electrical properties. In addition to measuring electrical conductivity, oxygen vacancy concentration was also evaluated using thermogravimetry, Neutron diffraction and Moessbauer Spectroscopy. The reducing conditions (CO{sub 2}/CO/H{sub 2} gas mixtures with steam) as encountered in a reactor environment can be expected to have significant influence on the mechanical properties of the oxides membranes. Various La based materials with and without Ti were selected as candidate membrane materials for OTM. The maximum electrical conductivity of LSF in air as a function of temperature was achieved at < 600 C and depends on the concentration of Sr (acceptor dopant). Oxygen occupancy in LSF was estimated using Neutron diffractometry and Moessbauer Spectroscopy by measuring magnetic moment changes depending on the Fe{sup 3+} and Fe{sup 4+} ratio. After extensive studies of candidate materials, lanthanum ferrites (LSF and LSFT) were selected as the favored materials for the oxygen transport membrane (OTM). LSF is a very good material for an OTM because of its high electronic and oxygen ionic conductivity if long term stability and mechanical strength are improved. LSFT not only exhibits p-type behavior in the high oxygen activity regime, but also has n-type conduction in reducing atmospheres. Higher concentrations of oxygen vacancies in the low oxygen activity regime may improve the performance of LSFT as an OTM. The hole concentration is related to the difference in the acceptor and donor concentration by the relation p = [Sr'{sub La}]-[Ti{sm_bullet}{sub Fe}]. The chemical formulation predicts that the hole concentration is, p = 0.8-0.45 or 0.35. Experimental measurements indicated that p is about {approx} 0.35. The activation energy of conduction is 0.2 eV which implies that LSCF conducts via the

  6. Oxygen Transport Ceramic Membranes

    SciTech Connect

    S. Bandopadhyay; N. Nagabhushana; Thomas W. Eagar; Harold R. Larson; Raymundo Arroyave; X.-D Zhou; Y.-W. Shin; H.U. Anderson; Nigel Browning; Alan Jacobson; C.A. Mims

    2003-11-01

    The present quarterly report describes some of the initial studies on newer compositions and also includes newer approaches to address various materials issues such as in metal-ceramic sealing. The current quarter's research has also focused on developing a comprehensive reliability model for predicting the structural behavior of the membranes in realistic conditions. In parallel to industry provided compositions, models membranes have been evaluated in varying environment. Of importance is the behavior of flaws and generation of new flaws aiding in fracture. Fracture mechanics parameters such as crack tip stresses are generated to characterize the influence of environment. Room temperature slow crack growth studies have also been initiated in industry provided compositions. The electrical conductivity and defect chemistry of an A site deficient compound (La{sub 0.55}Sr{sub 0.35}FeO{sub 3}) was studied. A higher conductivity was observed for La{sub 0.55}Sr{sub 0.35}FeO{sub 3} than that of La{sub 0.60}Sr{sub 0.40}FeO{sub 3} and La{sub 0.80}Sr{sub 0.20}FeO{sub 3}. Defect chemistry analysis showed that it was primarily contributed by a higher carrier concentration in La{sub 0.55}Sr{sub 0.35}FeO{sub 3}. Moreover, the ability for oxygen vacancy generation is much higher in La{sub 0.55}Sr{sub 0.35}FeO{sub 3} as well, which indicates a lower bonding strength between Fe-O and a possible higher catalytic activity for La{sub 0.55}Sr{sub 0.35}FeO{sub 3}. The program continued to investigate the thermodynamic properties (stability and phase separation behavior) and total conductivity of prototype membrane materials. The data are needed together with the kinetic information to develop a complete model for the membrane transport. Previous report listed initial measurements on a sample of La{sub 0.2}Sr{sub 0.8}Fe{sub 0.55}Ti{sub 0.45}O{sub 3-x} prepared in-house by Praxair. Subsequently, a second sample of powder from a larger batch of sample were characterized and compared with

  7. Electrically Conductive Porous Membrane

    NASA Technical Reports Server (NTRS)

    Burke, Kenneth Alan (Inventor)

    2014-01-01

    The present invention relates to an electrically conductive membrane that can be configured to be used in fuel cell systems to act as a hydrophilic water separator internal to the fuel cell, or as a water separator used with water vapor fed electrolysis cells, or as a water separator used with water vapor fed electrolysis cells, or as a capillary structure in a thin head pipe evaporator, or as a hydrophobic gas diffusion layer covering the fuel cell electrode surface in a fuel cell.

  8. Molecularly imprinted membranes.

    PubMed

    Trotta, Francesco; Biasizzo, Miriam; Caldera, Fabrizio

    2012-07-19

    Although the roots of molecularly imprinted polymers lie in the beginning of 1930s in the past century, they have had an exponential growth only 40-50 years later by the works of Wulff and especially by Mosbach. More recently, it was also proved that molecular imprinted membranes (i.e., polymer thin films) that show recognition properties at molecular level of the template molecule are used in their formation. Different procedures and potential application in separation processes and catalysis are reported. The influences of different parameters on the discrimination abilities are also discussed.

  9. Oxygen Transport Ceramic Membranes

    SciTech Connect

    S. Bandopadhyay; N. Nagabhushana; X.-D Zhou; Q. Cai; J. Yang; W.B. Yelon; W.J. James; H.U. Anderson; Alan Jacobson; C.A. Mims

    2004-05-01

    The present quarterly report describes some of the investigations on the structural properties of dense OTM bars provided by Praxair and studies on newer composition of Ti doped LSF. In this report, in situ neutron diffraction was used to characterize the chemical and structural properties of La{sub 0.2}Sr{sub 0.8}Fe{sub 0.55}Ti{sub 0.45}O{sub 3-{delta}} (here after as L2SF55T) specimen, which was subject to measurements of neutron diffraction from room temperature to 900 C. It was found that space group of R3c yielded a better refinement than a cubic structure of Pm3m. Oxygen occupancy was nearly 3 in the region from room temperature to 700 C, above which the occupancy decreased due to oxygen loss. Dense OTM bars provided by Praxair were loaded to fracture at varying stress rates. Studies were done at room temperature in air and at 1000 C in a specified environment to evaluate slow crack growth behavior. The X-Ray data and fracture mechanisms points to non-equilibrium decomposition of the LSFCO OTM membrane. The non-equilibrium conditions could probably be due to the nature of the applied stress field (stressing rates) and leads to transition in crystal structures and increased kinetics of decomposition. The formations of a Brownmillerite or Sr2Fe2O5 type structures, which are orthorhombic are attributed to the ordering of oxygen vacancies. The cubic to orthorhombic transitions leads to 2.6% increase in strains and thus residual stresses generated could influence the fracture behavior of the OTM membrane. Continued investigations on the thermodynamic properties (stability and phase-separation behavior) and total conductivity of prototype membrane materials were carried out. The data are needed together with the kinetic information to develop a complete model for the membrane transport. Previously characterization, stoichiometry and conductivity measurements for samples of La{sub 0.2}Sr{sub 0.8}Fe{sub 0.55}Ti{sub 0.45}O{sub 3-{delta}} were reported. In this report

  10. Molecularly Imprinted Membranes

    PubMed Central

    Trotta, Francesco; Biasizzo, Miriam; Caldera, Fabrizio

    2012-01-01

    Although the roots of molecularly imprinted polymers lie in the beginning of 1930s in the past century, they have had an exponential growth only 40–50 years later by the works of Wulff and especially by Mosbach. More recently, it was also proved that molecular imprinted membranes (i.e., polymer thin films) that show recognition properties at molecular level of the template molecule are used in their formation. Different procedures and potential application in separation processes and catalysis are reported. The influences of different parameters on the discrimination abilities are also discussed. PMID:24958291

  11. Supported liquid membranes

    SciTech Connect

    Danesi, P.R.

    1984-01-01

    The possibility of utilizing thin layers of organic solutions of solvent extraction reagents, immobilized on microporous inert supports interposed between two aqueous solutions, for selectively removing metal ions from a mixture represents an attractive alternative to liquid-liquid extraction. A detailed knowledge of the liquid-liquid extraction equilibria and mass transfer kinetics is required to understand and to describe quantitatively the rate laws which control the permeation of metal species through Supported Liquid Membranes (SLM) and to exploit them for separation processes. This paper attempts to understand the mechanism of transport through SLM.

  12. Protein-Induced Membrane Curvature Alters Local Membrane Tension

    PubMed Central

    Rangamani, Padmini; Mandadap, Kranthi K.; Oster, George

    2014-01-01

    Adsorption of proteins onto membranes can alter the local membrane curvature. This phenomenon has been observed in biological processes such as endocytosis, tubulation, and vesiculation. However, it is not clear how the local surface properties of the membrane, such as membrane tension, change in response to protein adsorption. In this article, we show that the partial differential equations arising from classical elastic model of lipid membranes, which account for simultaneous changes in shape and membrane tension due to protein adsorption in a local region, cannot be solved for nonaxisymmetric geometries using straightforward numerical techniques; instead, a viscous-elastic formulation is necessary to fully describe the system. Therefore, we develop a viscous-elastic model for inhomogeneous membranes of the Helfrich type. Using the newly available viscous-elastic model, we find that the lipids flow to accommodate changes in membrane curvature during protein adsorption. We show that, at the end of protein adsorption process, the system sustains a residual local tension to balance the difference between the actual mean curvature and the imposed spontaneous curvature. We also show that this change in membrane tension can have a functional impact such as altered response to pulling forces in the presence of proteins. PMID:25099814

  13. Proteins interacting with Membranes: Protein Sorting and Membrane Shaping

    NASA Astrophysics Data System (ADS)

    Callan-Jones, Andrew

    2015-03-01

    Membrane-bound transport in cells requires generating membrane curvature. In addition, transport is selective, in order to establish spatial gradients of membrane components in the cell. The mechanisms underlying cell membrane shaping by proteins and the influence of curvature on membrane composition are active areas of study in cell biophysics. In vitro approaches using Giant Unilamellar Vesicles (GUVs) are a useful tool to identify the physical mechanisms that drive sorting of membrane components and membrane shape change by proteins. I will present recent work on the curvature sensing and generation of IRSp53, a protein belonging to the BAR family, whose members, sharing a banana-shaped backbone, are involved in endocytosis. Pulling membrane tubes with 10-100 nm radii from GUVs containing encapsulated IRSp53 have, unexpectedly, revealed a non-monotonic dependence of the protein concentration on the tube as a function of curvature. Experiments also show that bound proteins alter the tube mechanics and that protein phase separation along the tube occurs at low tensions. I will present accompanying theoretical work that can explain these findings based on the competition between the protein's intrinsic curvature and the effective rigidity of a membrane-protein patch.

  14. Gas separations using inorganic membranes

    SciTech Connect

    Egan, B.Z.; Singh, S.P.N. ); Fain, D.E.; Roettger, G.E.; White, D.E. )

    1992-04-01

    This report summarizes the results from a research and development program to develop, fabricate, and evaluate inorganic membranes for separating gases at high temperatures and pressures in hostile process environments encountered in fossil energy conversion processes such as coal gasification. The primary emphasis of the research was on the separation and recovery of hydrogen from synthesis gas. Major aspects of the program included assessment of the worldwide research and development activity related to gas separations using inorganic membranes, identification and selection of candidate membrane materials, fabrication and characterization of membranes using porous membrane technology developed at the Oak Ridge K-25 Site, and evaluation of the separations capability of the fabricated membranes in terms of permeabilities and fluxes of gases.

  15. Catalytic Membrane Sensors

    SciTech Connect

    Boyle, T.J.; Brinker, C.J.; Gardner, T.J.; Hughes, R.C.; Sault, A.G.

    1998-12-01

    The proposed "catalytic membrane sensor" (CMS) was developed to generate a device which would selectively identify a specific reagent in a complex mixture of gases. This was to be accomplished by modifying an existing Hz sensor with a series of thin films. Through selectively sieving the desired component from a complex mixture and identifying it by decomposing it into Hz (and other by-products), a Hz sensor could then be used to detect the presence of the select component. The proposed "sandwich-type" modifications involved the deposition of a catalyst layered between two size selective sol-gel layers on a Pd/Ni resistive Hz sensor. The role of the catalyst was to convert organic materials to Hz and organic by-products. The role of the membraneo was to impart both chemical specificity by molecukir sieving of the analyte and converted product streams, as well as controlling access to the underlying Pd/Ni sensor. Ultimately, an array of these CMS elements encompassing different catalysts and membranes were to be developed which would enable improved selectivity and specificity from a compiex mixture of organic gases via pattern recognition methodologies. We have successfully generated a CMS device by a series of spin-coat deposited methods; however, it was determined that the high temperature required to activate the catalyst, destroys the sensor.

  16. Oxygen Transport Ceramic Membranes

    SciTech Connect

    S. Bandopadhyay; T. Nithyanantham; X.-D Zhou; Y-W. Sin; H.U. Anderson; Alan Jacobson; C.A. Mims

    2005-05-01

    The present quarterly report describes some of the investigations on the structural properties of dense OTM bars provided by Praxair and studies on newer composition of Ti doped Ti-substituted perovskites, La{sub 0.7}Sr{sub 0.3}Mn{sub 1-x}Ti{sub x}O{sub 3}, with 0 {le} x {le} 0.20, were investigated by neutron diffraction, magnetization, electric resistivity, and magnetoresistance (MR) measurements. All samples show a rhombohedral structure (space group R3C) from 10 K to room temperature. At room temperature, the cell parameters a, c and the unit cell volume increase with increasing Ti content. However, at 10 K, the cell parameter a has a maximum value for x = 0.10, and decreases for x > 0.10, while the unit cell volume remains nearly constant for x > 0.10. The average (Mn,Ti)-O bond length increases up to x = 0.15, and the (Mn,Ti)-O-(Mn,Ti) bond angle decreases with increasing Ti content to its minimum value at x = 0.15 at room temperature. Below the Curie temperature TC, the resistance exhibits metallic behavior for the x {le} 0.05 samples. A metal (semiconductor) to insulator transition is observed for the x {ge} 0.10 samples. A peak in resistivity appears below TC for all samples, and shifts to a lower temperature as x increases. The substitution of Mn by Ti decreases the 2p-3d hybridization between O and Mn ions, reduces the bandwidth W, and increases the electron-phonon coupling. Therefore, the TC shifts to a lower temperature and the resistivity increases with increasing Ti content. A field-induced shift of the resistivity maximum occurs at x {le} 0.10 compounds. The maximum MR effect is about 70% for La{sub 0.7}Sr{sub 0.3}Mn{sub 0.8}Ti{sub 0.2}O{sub 3}. The separation of TC and the resistivity maximum temperature T{sub {rho},max} enhances the MR effect in these compounds due to the weak coupling between the magnetic ordering and the resistivity as compared with La{sub 0.7}Sr{sub 0.3}MnO{sub 3}. The bulk densities of the membranes were determined using the

  17. Impact on floating membranes.

    PubMed

    Vandenberghe, Nicolas; Duchemin, Laurent

    2016-05-01

    When impacted by a rigid body, a thin elastic membrane with negligible bending rigidity floating on a liquid pool deforms. Two axisymmetric waves radiating from the impact point propagate. First, a longitudinal wave front, associated with in-plane deformation of the membrane and traveling at constant speed, separates an outward stress-free domain from a stretched domain. Then, in the stretched domain a dispersive transverse wave travels at a speed that depends on the local stretching rate. The dynamics is found to be self-similar in time. Using this property, we show that the wave dynamics is similar to the capillary waves that propagate at a liquid-gas interface but with a surface tension coefficient that depends on impact speed. During wave propagation, we observe the development of a buckling instability that gives rise to radial wrinkles. We address the dynamics of this fluid-body system, including the rapid deceleration of an impactor of finite mass, an issue that may have applications in the domain of absorption of impact energy. PMID:27300958

  18. Supported liquid membrane electrochemical separators

    DOEpatents

    Pemsler, J. Paul; Dempsey, Michael D.

    1986-01-01

    Supported liquid membrane separators improve the flexibility, efficiency and service life of electrochemical cells for a variety of applications. In the field of electrochemical storage, an alkaline secondary battery with improved service life is described in which a supported liquid membrane is interposed between the positive and negative electrodes. The supported liquid membranes of this invention can be used in energy production and storage systems, electrosynthesis systems, and in systems for the electrowinning and electrorefining of metals.

  19. When plasmonics meets membrane technology

    NASA Astrophysics Data System (ADS)

    Politano, A.; Cupolillo, A.; Di Profio, G.; Arafat, H. A.; Chiarello, G.; Curcio, E.

    2016-09-01

    In this review, we present the applications of thermoplasmonics in membrane processes. We discuss the influence of the heat capacity of the solvent, the amount of plasmonic nanoparticles in the membrane, the intensity of the light source and the transmembrane flow rate on the increase of permeability. Remarkably, thermoplasmonic effects do not involve any noticeable loss of membrane rejection. Herein, we consider application feasibilities, including application fields, requirements of feed, alternatives of light sources, promising thermoplasmonic nanoparticles and scaling up issues.

  20. When plasmonics meets membrane technology.

    PubMed

    Politano, A; Cupolillo, A; Di Profio, G; Arafat, H A; Chiarello, G; Curcio, E

    2016-09-14

    In this review, we present the applications of thermoplasmonics in membrane processes. We discuss the influence of the heat capacity of the solvent, the amount of plasmonic nanoparticles in the membrane, the intensity of the light source and the transmembrane flow rate on the increase of permeability. Remarkably, thermoplasmonic effects do not involve any noticeable loss of membrane rejection. Herein, we consider application feasibilities, including application fields, requirements of feed, alternatives of light sources, promising thermoplasmonic nanoparticles and scaling up issues.