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Sample records for membrane deformations activate

  1. Versatile membrane deformation potential of activated pacsin.

    Directory of Open Access Journals (Sweden)

    Shih Lin Goh

    Full Text Available Endocytosis is a fundamental process in signaling and membrane trafficking. The formation of vesicles at the plasma membrane is mediated by the G protein dynamin that catalyzes the final fission step, the actin cytoskeleton, and proteins that sense or induce membrane curvature. One such protein, the F-BAR domain-containing protein pacsin, contributes to this process and has been shown to induce a spectrum of membrane morphologies, including tubules and tube constrictions in vitro. Full-length pacsin isoform 1 (pacsin-1 has reduced activity compared to its isolated F-BAR domain, implicating an inhibitory role for its C-terminal Src homology 3 (SH3 domain. Here we show that the autoinhibitory, intramolecular interactions in pacsin-1 can be released upon binding to the entire proline-rich domain (PRD of dynamin-1, resulting in potent membrane deformation activity that is distinct from the isolated F-BAR domain. Most strikingly, we observe the generation of small, homogenous vesicles with the activated protein complex under certain experimental conditions. In addition, liposomes prepared with different methods yield distinct membrane deformation morphologies of BAR domain proteins and apparent activation barriers to pacsin-1's activity. Theoretical free energy calculations suggest bimodality of the protein-membrane system as a possible source for the different outcomes, which could account for the coexistence of energetically equivalent membrane structures induced by BAR domain-containing proteins in vitro. Taken together, our results suggest a versatile role for pacsin-1 in sculpting cellular membranes that is likely dependent both on protein structure and membrane properties.

  2. Local membrane deformations activate Ca2+-dependent K+ and anionic currents in intact human red blood cells

    DEFF Research Database (Denmark)

    Dyrda, Agnieszka; Cytlak, Urszula; Ciuraszkiewicz, Anna

    2010-01-01

    by such flow, as well as the local membrane deformations generated in certain pathological conditions, such as sickle cell anemia, have been shown to increase membrane permeability, based largely on experimentation with red cell suspensions. We attempted here the first measurements of membrane currents......-activated transient PCa observed here under local membrane deformation is a likely contributor to the Ca(2+)-mediated effects observed during the normal aging process of red blood cells, and to the increased Ca(2+) content of red cells in certain hereditary anemias such as thalassemia and sickle cell anemia....

  3. Local membrane deformations activate Ca2+-dependent K+ and anionic currents in intact human red blood cells.

    Directory of Open Access Journals (Sweden)

    Agnieszka Dyrda

    Full Text Available BACKGROUND: The mechanical, rheological and shape properties of red blood cells are determined by their cortical cytoskeleton, evolutionarily optimized to provide the dynamic deformability required for flow through capillaries much narrower than the cell's diameter. The shear stress induced by such flow, as well as the local membrane deformations generated in certain pathological conditions, such as sickle cell anemia, have been shown to increase membrane permeability, based largely on experimentation with red cell suspensions. We attempted here the first measurements of membrane currents activated by a local and controlled membrane deformation in single red blood cells under on-cell patch clamp to define the nature of the stretch-activated currents. METHODOLOGY/PRINCIPAL FINDINGS: The cell-attached configuration of the patch-clamp technique was used to allow recordings of single channel activity in intact red blood cells. Gigaohm seal formation was obtained with and without membrane deformation. Deformation was induced by the application of a negative pressure pulse of 10 mmHg for less than 5 s. Currents were only detected when the membrane was seen domed under negative pressure within the patch-pipette. K(+ and Cl(- currents were strictly dependent on the presence of Ca(2+. The Ca(2+-dependent currents were transient, with typical decay half-times of about 5-10 min, suggesting the spontaneous inactivation of a stretch-activated Ca(2+ permeability (PCa. These results indicate that local membrane deformations can transiently activate a Ca(2+ permeability pathway leading to increased [Ca(2+](i, secondary activation of Ca(2+-sensitive K(+ channels (Gardos channel, IK1, KCa3.1, and hyperpolarization-induced anion currents. CONCLUSIONS/SIGNIFICANCE: The stretch-activated transient PCa observed here under local membrane deformation is a likely contributor to the Ca(2+-mediated effects observed during the normal aging process of red blood cells, and

  4. Axisymmetric finite deformation membrane problems

    Energy Technology Data Exchange (ETDEWEB)

    Feng, W.W.

    1980-12-12

    Many biomechanic problems involve the analysis of finite deformation axisymmetric membranes. This paper presents the general formulation for solving a class of axisymmetric membrane problems. The material nonlinearity, as well as the geometric nonlinearity, is considered. Two methods are presented to solve these problems. The first method is solving a set of differential equilibrium equations. The governing equations are reduced to three first-order ordinary-differential equations with explicit derivatives. The second method is the Ritz method where a general potential energy functional valid for all axisymmetric deformed positions is presented. The geometric admissible functions that govern the deformed configuration are written in terms of a series with unknown coefficients. These unknown coefficients are determined by the minimum potential energy principle that of all geometric admissible deformed configurations, the equilibrium configuration minimizes the potential energy. Some examples are presented. A comparison between these two methods is mentioned.

  5. Corrugated Membrane Nonlinear Deformation Process Calculation

    Directory of Open Access Journals (Sweden)

    A. S. Nikolaeva

    2015-01-01

    Full Text Available Elastic elements are widely used in instrumentation. They are used to create a particular interference between the parts, for accumulating mechanical energy, as the motion transmission elements, elastic supports, and sensing elements of measuring devices. Device reliability and quality depend on the calculation accuracy of the elastic elements. A corrugated membrane is rather common embodiment of the elastic element.The corrugated membrane properties depend largely on its profile i.e. a generatrix of the meridian surface.Unlike other types of pressure elastic members (bellows, tube spring, the elastic characteristics of which are close to linear, an elastic characteristic of the corrugated membrane (typical movement versus external load is nonlinear. Therefore, the corrugated membranes can be used to measure quantities, nonlinearly related to the pressure (e.g., aircraft air speed, its altitude, pipeline fluid or gas flow rate. Another feature of the corrugated membrane is that significant movements are possible within the elastic material state. However, a significant non-linearity of membrane characteristics leads to severe complicated calculation.This article is aimed at calculating the corrugated membrane to obtain the elastic characteristics and the deformed shape of the membrane meridian, as well as at investigating the processes of buckling. As the calculation model, a thin-walled axisymmetric shell rotation is assumed. The material properties are linearly elastic. We consider a corrugated membrane of sinusoidal profile. The membrane load is a uniform pressure.The algorithm for calculating the mathematical model of an axisymmetric corrugated membrane of constant thickness, based on the Reissner’s theory of elastic thin shells, was realized as the author's program in C language. To solve the nonlinear problem were used a method of changing the subspace of control parameters, developed by S.S., Gavriushin, and a parameter marching method

  6. Distributed microscopic actuation analysis of deformable plate membrane mirrors

    Science.gov (United States)

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

    2018-02-01

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

  7. Large Deformation Mechanics of Plasma Membrane Chained Vesicles in Cells

    Science.gov (United States)

    Kosawada, Tadashi; Sanada, Kouichi; Takano, Tetsuo

    The clathrin-coated pits, vesicles and chained vesicles on the inner surface of the plasma membrane facilitate the cell to transport specific extracellular macromolecules. This cellular process is strongly involved with large mechanical deformations of the plasma membrane accompanied by changes in membrane curvature. The assembly of the clathrin coat is thought to provide curvature into the membrane. Hence, effects of in-plane shear elasticity due to these coat structure may be significant on the vesicular mechanics. In this study, large deformation mechanics of plasma membrane chained vesicles in cells have been formulated based on minimization of bending and in-plane shear strain energy of the membrane. Effects of outer surrounding cytoplasmic flat membrane upon mechanically stable shapes of the vesicles were revealed, while effects of in-plane shear elasticity were partly discussed.

  8. Large Deformation and Adhesive Contact Studies of Axisymmetric Membranes

    Science.gov (United States)

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

    2013-01-01

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

  9. Large deformation and adhesive contact studies of axisymmetric membranes.

    Science.gov (United States)

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

    2013-02-05

    A model membrane contact system consisting of an acrylic copolymer membrane and a PDMS substrate was utilized to evaluate a recently developed nonlinear large-deformation adhesive contact analysis. Direct measurements of the local membrane apex strain during noncontact inflation indicated that the neo-Hookean model provides an accurate measure of membrane strain and supports its use as the strain energy function for the analysis. Two membrane contact geometries, exhibiting significantly different strain distributions during withdrawal, were investigated. The first examines the wet contact of an air pressurized membrane. The second looks at the dry contact of a fluid deformed membrane in which a stepper motor controls membrane-substrate separation. A time-dependent modulus emerges from the analysis, with principal tensions obtained from a comparison of predicted and experimental membrane profiles. The applicability of this numerical analysis for determining membrane tension, however, is limited by wrinkling instabilities and viscoelasticity. For this reason, a conceptually simpler method, based on the direct measurement of the membrane tension and contact angle, was also utilized. The traditional peel energy defined with this direct measurement accurately described the membrane/substrate adhesive interactions, giving well-defined peel energies that were independent of the detailed strain state of the membrane.

  10. [Role of protein kinases of human red cell membrane in deformability and aggregation changes].

    Science.gov (United States)

    Murav'ev, A V; Maĭmistova, A A; Tikhomirova, I A; Bulaeva, S V; Mikhaĭlov, P V; Murav'ev, A A

    2012-01-01

    The proteomic analysis has showed that red cell membrane contains several kinases and phosphatases. Therefore the aim of this study was to investigate the role of protein kinases of human red cell membrane in deformability and aggregation changes. Exposure of red blood cells (RBCs) to some chemical compounds led to change in the RBC microrheological properties. When forskolin (10 microM), an adenylyl cyclase (AC) and a protein kinase A (PKA) stimulator was added to RBC suspension, the RBC deformability (RBCD) was increased by 20% (p RBCA) was significantly decreased under these conditions (p RBCA lowering. The similar effect was found when cells were incubated with cisplatin as a tyrosine protein kinase (TPK) activator. It is important to note that a selective TPK inhibitor--lavendustin eliminated the above mention effects. On the whole the total data clearly show that the red cell aggregation and deformation changes were connected with an activation of the different intracellular signaling pathways.

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

    Science.gov (United States)

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

    2018-03-01

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

  12. Tensile deformation of polytetrafluoroethylene hollow fiber membranes used for water purification.

    Science.gov (United States)

    Yonezu, Akio; Iio, Shouichi; Itonaga, Takehiro; Yamamura, Hiroshi; Chen, Xi

    2014-01-01

    The tensile deformation behavior of polytetrafluoroethylene (PTFE) hollow fiber membranes is studied. PTFE membranes at present have sub-micron pores with an open cell structure, which plays a critical role in water purification. One of the main challenges in water purification is that the pore structure becomes covered with biofouling, leading to blocked pores. To maintain the capacity for water purification, physical cleaning along with mechanical deformation is usually conducted. Thus, it is crucial to understand the mechanical properties, in particular the deformation behavior, of the membrane fibers. Using uniaxial tension experiments, we established a fundamental discrete model to describe the deformation behavior of a porous structure using a finite element method. The present model enables the prediction of the macroscopic deformation behavior of the membrane, by taking into account the changes of pore structure. The insight may be useful for porous membrane fabrication and provide insights for the reliable operation of water purification.

  13. Habitual Physical Activity, Peripheral Neuropathy, Foot Deformities ...

    African Journals Online (AJOL)

    Results: Habitual physical activity index (3.2 ± 0.83) was highest in work-related activities; 69 (26.1 %) patients presented with peripheral neuropathy and 52 (19. 7%) had the lowest limb function. Pes planus was the most prevalent foot deformity (20.1%). Significant differences existed in physical activity indices across ...

  14. Deformations of closed strings and topological open membranes

    NARCIS (Netherlands)

    Hofman, C.; Ma, W.K.

    2001-01-01

    We study deformations of topological closed strings. A well-known example is the perturba- tion of a topological closed string by itself, where the associative OPE product is deformed, and which isgoverned by the WD VV equations. Our main in terest will be closed strings that arise as the

  15. Deformations of closed strings and topological open membranes

    NARCIS (Netherlands)

    Hofman, C.

    We study deformations of topological closed strings. A well-known example is the perturbation of a topological closed string by itself, where the associative OPE product is deformed, and which is governed by the WDVV equations. Our main interest will be closed strings that arise as the boundary

  16. Modelling Protein-induced Membrane Deformation using Monte Carlo and Langevin Dynamics Simulations

    Science.gov (United States)

    Radhakrishnan, R.; Agrawal, N.; Ramakrishnan, N.; Kumar, P. B. Sunil; Liu, J.

    2010-11-01

    In eukaryotic cells, internalization of extracellular cargo via the cellular process of endocytosis is orchestrated by a variety of proteins, many of which are implicated in membrane deformation/bending. We model the energetics of deformations membranes by using the Helfrich Hamiltonian using two different formalisms: (i) Cartesian or Monge Gauge using Langevin dynamics; (ii) Curvilinear coordinate system using Monte Carlo (MC). Monge gauge approach which has been extensively studied is limited to small deformations of the membrane and cannot describe extreme deformations. Curvilinear coordinate approach can handle large deformation limits as well as finite-temperature membrane fluctuations; here we employ an unstructured triangular mesh to compute the local curvature tensor, and we evolve the membrane surface using a MC method. In our application, we compare the two approaches (i and ii above) to study how the spatial assembly of curvature inducing proteins leads to vesicle budding from a planar membrane. We also quantify how the curvature field of the membrane impacts the spatial segregation of proteins.

  17. Modeling electrically active viscoelastic membranes.

    Directory of Open Access Journals (Sweden)

    Sitikantha Roy

    Full Text Available The membrane protein prestin is native to the cochlear outer hair cell that is crucial to the ear's amplification and frequency selectivity throughout the whole acoustic frequency range. The outer hair cell exhibits interrelated dimensional changes, force generation, and electric charge transfer. Cells transfected with prestin acquire unique active properties similar to those in the native cell that have also been useful in understanding the process. Here we propose a model describing the major electromechanical features of such active membranes. The model derived from thermodynamic principles is in the form of integral relationships between the history of voltage and membrane resultants as independent variables and the charge density and strains as dependent variables. The proposed model is applied to the analysis of an active force produced by the outer hair cell in response to a harmonic electric field. Our analysis reveals the mechanism of the outer hair cell active (isometric force having an almost constant amplitude and phase up to 80 kHz. We found that the frequency-invariance of the force is a result of interplay between the electrical filtering associated with prestin and power law viscoelasticity of the surrounding membrane. Paradoxically, the membrane viscoelasticity boosts the force balancing the electrical filtering effect. We also consider various modes of electromechanical coupling in membrane with prestin associated with mechanical perturbations in the cell. We consider pressure or strains applied step-wise or at a constant rate and compute the time course of the resulting electric charge. The results obtained here are important for the analysis of electromechanical properties of membranes, cells, and biological materials as well as for a better understanding of the mechanism of hearing and the role of the protein prestin in this mechanism.

  18. Wrapping of a deformable nanoparticle by the cell membrane: Insights into the flexibility-regulated nanoparticle-membrane interaction

    Science.gov (United States)

    Tang, Huayuan; Zhang, Hongwu; Ye, Hongfei; Zheng, Yonggang

    2016-09-01

    Although many researches have been conducted on the interaction of the cell membrane with the rigid nanoparticle (NP), relatively little is known about the interaction of the membrane with the deformable NP, which is a promising kind of drug delivery carrier. In this paper, we investigate the wrapping of a deformable NP by the membrane, with particular attention paid to the location of the NP. Phase diagrams with respect to the normalized NP-membrane adhesion strength and the bending stiffness ratio between the NP and membrane are presented. The results show that the NP is easier to be fully wrapped but harder to be shallowly wrapped when the NP locates outside than inside the vesicle. For the system with an outside NP, there are three distinct stages separated by two critical bending stiffness ratios as the NP becomes softer. Moreover, the critical normalized adhesion strength required for a deformable NP to be fully wrapped is the same as that for a rigid NP when the bending stiffness ratio is higher than a critical value, which is different from the wrapping behavior by an initially flat membrane. In addition, a larger vesicle size facilitates the full wrapping configuration when the NP is inside, whereas it prohibits it when the NP is outside. These results are consistent with the previous research and can provide guidelines for the design of drug delivery systems based on the flexibility-tunable NPs.

  19. Liposomal internal viscosity affects the fate of membrane deformation induced by hypertonic treatment.

    Science.gov (United States)

    Fujiwara, Kei; Yanagisawa, Miho

    2017-12-13

    Artificial lipid membranes have been utilized to understand the physical mechanisms of the deformation patterns of live cells. However, typical artificial membrane systems contain only dilute components compared to those in the cytoplasm of live cells. By using giant unilamellar liposomes containing dense protein solutions similar to those in live cells, we here reveal that viscosity derived from internal crowding affects the deformation patterns of lipid membranes. After hypertonic treatment, liposome deformation patterns transitioned from budding to tubing when the initial internal macromolecular concentrations were increased. Remarkably, instead of observing different transition concentrations between two species of macromolecules, the viscosity at the transition concentration was found to be similar. Further analyses clearly demonstrated that the internal viscosity affects the deformation patterns of lipid membranes induced by hypertonic treatment. These results indicate that the viscosity of the cytoplasm is a key factor in determining cell deformation, and suggest the association of a process involving dynamic instability, such as a viscous fingering phenomenon, during the determination of deformation patterns by hypertonic treatment.

  20. Hybrid Electrostatic/Flextensional Deformable Membrane Mirror for Lightweight, Large Aperture and Cryogenic Space Telescopes, Phase II

    Data.gov (United States)

    National Aeronautics and Space Administration — TRS Technologies proposes innovative hybrid electrostatic/flextensional membrane deformable mirror capable of large amplitude aberration correction for large...

  1. Filament networks attached to membranes: cytoskeletal pressure and local bilayer deformation

    Energy Technology Data Exchange (ETDEWEB)

    Auth, Thorsten [Department of Materials and Interfaces, Weizmann Institute of Science, PO Box 26, Rehovot 76100 (Israel); Safran, S A [Department of Materials and Interfaces, Weizmann Institute of Science, PO Box 26, Rehovot 76100 (Israel); Gov, Nir S [Department of Chemical Physics, Weizmann Institute of Science, PO Box 26, Rehovot 76100 (Israel)

    2007-11-15

    Several cell types, among them red blood cells, have a cortical, two-dimensional (2D) network of filaments sparsely attached to their lipid bilayer. In many mammalian cells, this 2D polymer network is connected to an underlying 3D, more rigid cytoskeleton. In this paper, we consider the pressure exerted by the thermally fluctuating, cortical network of filaments on the bilayer and predict the bilayer deformations that are induced by this pressure. We treat the filaments as flexible polymers and calculate the pressure that a network of such linear chains exerts on the bilayer; we then minimize the bilayer shape in order to predict the resulting local deformations. We compare our predictions with membrane deformations observed in electron micrographs of red blood cells. The polymer pressure along with the resulting membrane deformation can lead to compartmentalization, regulate in-plane diffusion and may influence protein sorting as well as transmit signals to the polymerization of the underlying 3D cytoskeleton.

  2. Filament networks attached to membranes: cytoskeletal pressure and local bilayer deformation

    International Nuclear Information System (INIS)

    Auth, Thorsten; Safran, S A; Gov, Nir S

    2007-01-01

    Several cell types, among them red blood cells, have a cortical, two-dimensional (2D) network of filaments sparsely attached to their lipid bilayer. In many mammalian cells, this 2D polymer network is connected to an underlying 3D, more rigid cytoskeleton. In this paper, we consider the pressure exerted by the thermally fluctuating, cortical network of filaments on the bilayer and predict the bilayer deformations that are induced by this pressure. We treat the filaments as flexible polymers and calculate the pressure that a network of such linear chains exerts on the bilayer; we then minimize the bilayer shape in order to predict the resulting local deformations. We compare our predictions with membrane deformations observed in electron micrographs of red blood cells. The polymer pressure along with the resulting membrane deformation can lead to compartmentalization, regulate in-plane diffusion and may influence protein sorting as well as transmit signals to the polymerization of the underlying 3D cytoskeleton

  3. Active Beam Shaping System and Method Using Sequential Deformable Mirrors

    Science.gov (United States)

    Norman, Colin A. (Inventor); Pueyo, Laurent A. (Inventor)

    2015-01-01

    An active optical beam shaping system includes a first deformable mirror arranged to at least partially intercept an entrance beam of light and to provide a first reflected beam of light, a second deformable mirror arranged to at least partially intercept the first reflected beam of light from the first deformable mirror and to provide a second reflected beam of light, and a signal processing and control system configured to communicate with the first and second deformable mirrors. The first deformable mirror, the second deformable mirror and the signal processing and control system together provide a large amplitude light modulation range to provide an actively shaped optical beam.

  4. Olopatadine Inhibits Exocytosis in Rat Peritoneal Mast Cells by Counteracting Membrane Surface Deformation

    Directory of Open Access Journals (Sweden)

    Asuka Baba

    2015-01-01

    Full Text Available Backgroud/Aims: Besides its anti-allergic properties as a histamine receptor antagonist, olopatadine stabilizes mast cells by inhibiting the release of chemokines. Since olopatadine bears amphiphilic features and is preferentially partitioned into the lipid bilayers of the plasma membrane, it would induce some morphological changes in mast cells and thus affect the process of exocytosis. Methods: Employing the standard patch-clamp whole-cell recording technique, we examined the effects of olopatadine and other anti-allergic drugs on the membrane capacitance (Cm in rat peritoneal mast cells during exocytosis. Using confocal imaging of a water-soluble fluorescent dye, lucifer yellow, we also examined their effects on the deformation of the plasma membrane. Results: Low concentrations of olopatadine (1 or 10 µM did not significantly affect the GTP-γ-S-induced increase in the Cm. However, 100 µM and 1 mM olopatadine almost totally suppressed the increase in the Cm. Additionally, these doses completely washed out the trapping of the dye on the cell surface, indicating that olopatadine counteracted the membrane surface deformation induced by exocytosis. As shown by electron microscopy, olopatadine generated inward membrane bending in mast cells. Conclusion: This study provides electrophysiological evidence for the first time that olopatadine dose-dependently inhibits the process of exocytosis in rat peritoneal mast cells. Such mast cell stabilizing properties of olopatadine may be attributed to its counteracting effects on the plasma membrane deformation in degranulating mast cells.

  5. Self-positioning of polymer membranes driven by thermomechanically induced plastic deformation

    DEFF Research Database (Denmark)

    Häfliger, Daniel; Hansen, Ole; Boisen, Anja

    2006-01-01

    Stress in polymeric resins is tailored by a thermomechanical process. It allows for controlled self-positioning of membranes in microdevices (see Figure). The process makes specific use of plastic deformation that results from the low viscosity of the polymer. This demonstrates that polymers offer...

  6. Active deformation offshore the Western Transverse Ranges

    Science.gov (United States)

    Ucarkus, G.; Driscoll, N. W.; Brothers, D. S.; Kent, G.; Rockwell, T. K.

    2014-12-01

    The Transverse Ranges within the structural province of southern California, an east-west trending active fold and thrust belt system, has rapid uplift rates that are capable of generating large earthquakes and tsunamis. This system to the west consists of north and south dipping reverse faults offshore Santa Barbara and Ventura (i.e., Pitas Point fault, Red Mountain fault, Rincon Creek fault). Ventura Avenue Anticline (VAA) is one of the fastest uplifting structure within this system has experienced nearly 2.7 km of structural uplift since fold initiation about 200-300 thousand years ago, yielding an average uplift rate of 9-13 mm/yr. Mapped and dated Holocene marine terraces between Ventura and Carpenteria reveal that large uplift events occurred at 0.8 ka and 1.9 ka; a recurrence interval of approximately a thousand years. The VAA trends offshore to the west and is buried by sediment from Rincon Creek. This sediment completely obscures the surficial expression of the fold between Rincon Point and Punta Gorda, indicating that Holocene sedimentation has kept pace with fold growth. Given the high sedimentation rate, each uplift event should be captured by stratigraphic rotation and onlap, and formation of angular unconformities. With that perspective, we acquired ~240 km-long very high-resolution (decimeter) CHIRP seismic reflection data from offshore Santa Barbara in the west to Ventura in the east, in order to examine discrete folding/uplift events that are preserved in the Holocene sediment record. CHIRP data together with re-processed USGS sparker profiles provide new constraints on timing and architecture of deformation offshore. A transgressive surface that dates back to ~9.5 kyr B.P is identified in seismic reflection data and dips landward; bending of the transgressive surface appears to be due to active folding and faulting. Observed onlapping sediments together with the deformation of the transgressive surface mark the onset of deformation while periods

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

    Science.gov (United States)

    Yao, Lingxing; Mori, Yoichiro

    2017-12-01

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

  8. CRC 1114 - Report Membrane Deformation by N-BAR Proteins: Extraction of membrane geometry and protein diffusion characteristics from MD simulations

    OpenAIRE

    Peters, Jan Henning; Gräser, Carsten; Klein, Rupert

    2017-01-01

    We describe simulations of Proteins and artificial pseudo-molecules interacting and shaping lipid bilayer membranes. We extract protein diffusion Parameters, membrane deformation profiles and the elastic properties of the used membrane models in preparation of calculations based on a large scale continuum model.

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

    KAUST Repository

    Zhang, Xiaoyuan

    2010-03-01

    Cation (CEMs) and anion exchange membrane (AEMs) are commonly used in microbial fuel cells (MFCs) to enhance Coulombic efficiencies (CEs) by reducing thefluxof oxygen through the cathode to bacteriaonthe anode. AEMs typically work better than CEMs, but in initial experiments we observed the opposite using a membrane electrode assembly MFC. The reason was identified to be membrane deformation, which resulted in water and gas trapped between the membrane and cathode. To correct this, stainless steel mesh was used to press the membrane flat against the cathode. With the steel mesh, AEM performance increased to 46±4W/m3 in a single cathode MFC, and 98±14W/m3 in a double-cathode MFC. These power densities were higher than those using a CEM of 32±2W/m3 (single cathode) and 63±6W/m3 (double cathode). Higher pH gradients across the membrane and salt precipitation on the cathode were responsible for the reduced performance of the CEM compared to the AEM. CEs reached over 90% for both membranes at >2A/m2. These results demonstrate the importance of avoiding water accumulation in thin films between membranes and electrodes, and explain additional reasons for poorer performance of CEMs compared to AEMs. © 2009 Elsevier B.V.

  10. Dynamics of Deformable Active Particles under External Flow Field

    Science.gov (United States)

    Tarama, Mitsusuke

    2017-10-01

    In most practical situations, active particles are affected by their environment, for example, by a chemical concentration gradient, light intensity, gravity, or confinement. In particular, the effect of an external flow field is important for particles swimming in a solvent fluid. For deformable active particles such as self-propelled liquid droplets and active vesicles, as well as microorganisms such as euglenas and neutrophils, a general description has been developed by focusing on shape deformation. In this review, we present our recent studies concerning the dynamics of a single active deformable particle under an external flow field. First, a set of model equations of active deformable particles including the effect of a general external flow is introduced. Then, the dynamics under two specific flow profiles is discussed: a linear shear flow, as the simplest example, and a swirl flow. In the latter case, the scattering dynamics of the active deformable particles by the swirl flow is also considered.

  11. Activation of interfacial enzymes at membrane surfaces

    DEFF Research Database (Denmark)

    Mouritsen, Ole G.; Andresen, Thomas Lars; Halperin, Avi

    2006-01-01

    A host of water-soluble enzymes are active at membrane surfaces and in association with membranes. Some of these enzymes are involved in signalling and in modification and remodelling of the membranes. A special class of enzymes, the phospholipases, and in particular secretory phospholipase A2 (s...

  12. Free Vibrations of a Nonlinearly Deformable Isotropic on the Average Composite Rectangular Membrane

    Science.gov (United States)

    Tarasyuk, I. A.; Kravchuk, A. S.; Mikhasev, G. I.

    2018-03-01

    A refined vibration equation of a rectangular membrane is derived in this paper. It allows determining the natural frequencies as functions of the mechanical characteristics of an asymmetrically stretched membrane. The dynamic equation is generalized to the case of a nonlinearly deformable isotropic on the average composite material. An approximate analytical solution of the problem is found employing a new homogenization technique. This method is based on estimation of the effective deformation characteristics of the composite material. The range of its effective characteristics is obtained from the rule of mixtures for the stresses and strains found assuming Voigt and Reuss hypotheses. The nonlinear behavior of the material is modeled using the bilinear Prandtl diagrams as constitutive equations for components of the composite. The effective elastic moduli, hardening modulus, yield stress, and the natural frequencies as functions of elastoplastic characteristics of the composite are obtained analytically in a closed form.

  13. The F-BAR domains from srGAP1, srGAP2 and srGAP3 regulate membrane deformation differently

    Science.gov (United States)

    Coutinho-Budd, Jaeda; Ghukasyan, Vladimir; Zylka, Mark J.; Polleux, Franck

    2012-01-01

    Summary Coordination of membrane deformation and cytoskeletal dynamics lies at the heart of many biological processes critical for cell polarity, motility and morphogenesis. We have recently shown that Slit-Robo GTPase-activating protein 2 (srGAP2) regulates neuronal morphogenesis through the ability of its F-BAR domain to regulate membrane deformation and induce filopodia formation. Here, we demonstrate that the F-BAR domains of two closely related family members, srGAP1 and srGAP3 [designated F-BAR(1) and F-BAR(3), respectively] display significantly different membrane deformation properties in non-neuronal COS7 cells and in cortical neurons. F-BAR(3) induces filopodia in both cell types, though less potently than F-BAR(2), whereas F-BAR(1) prevents filopodia formation in cortical neurons and reduces plasma membrane dynamics. These three F-BAR domains can heterodimerize, and they act synergistically towards filopodia induction in COS7 cells. As measured by fluorescence recovery after photobleaching, F-BAR(2) displays faster molecular dynamics than F-BAR(3) and F-BAR(1) at the plasma membrane, which correlates well with its increased potency to induce filopodia. We also show that the molecular dynamic properties of F-BAR(2) at the membrane are partially dependent on F-Actin. Interestingly, acute phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] depletion in cells does not interfere with plasma membrane localization of F-BAR(2), which is compatible with our result showing that F-BAR(2) binds to a broad range of negatively-charged phospholipids present at the plasma membrane, including phosphatidylserine (PtdSer). Overall, our results provide novel insights into the functional diversity of the membrane deformation properties of this subclass of F-BAR-domains required for cell morphogenesis. PMID:22467852

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

    Science.gov (United States)

    Yoon, Youngdae; Zhang, Xiuqi; Cho, Wonhwa

    2012-01-01

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

  15. Cyclic mechanical deformation stimulates human lung fibroblast proliferation and autocrine growth factor activity.

    Science.gov (United States)

    Bishop, J E; Mitchell, J J; Absher, P M; Baldor, L; Geller, H A; Woodcock-Mitchell, J; Hamblin, M J; Vacek, P; Low, R B

    1993-08-01

    Cellular hypertrophy and hyperplasia and increased extracellular matrix deposition are features of tissue hypertrophy resulting from increased work load. It is known, for example, that mechanical forces play a critical role in lung development, cardiovascular remodeling following pressure overload, and skeletal muscle growth. The mechanisms involved in these processes, however, remain unclear. Here we examined the effect of mechanical deformation on fibroblast function in vitro. IMR-90 human fetal lung fibroblasts grown on collagen-coated silastic membranes were subjected to cyclical mechanical deformation (10% increase in culture surface area; 1 Hz) for up to 5 days. Cell number was increased by 39% after 2 days of deformation (1.43 +/- .01 x 10(5) cells/membrane compared with control, 1.03 +/- 0.02 x 10(5) cells; mean +/- SEM; P < 0.02) increasing to 163% above control by 4 days (2.16 +/- 0.16 x 10(5) cells compared with 0.82 +/- 0.03 x 10(5) cells; P < 0.001). The medium from mechanically deformed cells was mitogenic for IMR-90 cells, with maximal activity in the medium from cells mechanically deformed for 2 days (stimulating cell replication by 35% compared with media control; P < 0.002). These data suggest that mechanical deformation stimulates human lung fibroblast replication and that this effect is mediated by the release of autocrine growth factors.

  16. Relationship between water activity, deformation speed, and crispness characterization

    NARCIS (Netherlands)

    Castro Prada, E.M.; Primo Martin, C.; Meinders, M.B.J.; Hamer, R.J.; Vliet, van T.

    2009-01-01

    Very little is known on the rate dependency of the fracture behavior of crispy products as a function of water activity (Aw). Therefore, the effect of deformation speed on instrumental and sensory crispness was studied as a function of Aw. Deformation speed clearly affects the transition Aw range

  17. AN ACTIVE VALVE WITH A CLAMPED MEMBRANE

    DEFF Research Database (Denmark)

    2009-01-01

    An active valve for use e.g. in fluidic microsystems is provided, wherein the active valve comprises a membrane having at least one flow gate, arranged between a first and a second substantially rigid element. Adjusting means provides an adjustment of the clamping force on membrane arranged between...

  18. Habitual physical activity, peripheral neuropathy, foot deformities ...

    African Journals Online (AJOL)

    Function Scale, and a self-designed foot deformity audit form. Results: Habitual ... glycaemic control including the general control of the disorder ...... Intern Med. 2005;20(3):259–66. http://dx.doi.org/10.1111/j.1525-. 1497.2005.40198.x. 43. Gutiérrez-Fisac JL, Guallar-Castillón P, Díez-Gañán L, et al. Work-related physical ...

  19. Anti-deformed Polyacrylonitrile/Polysulfone Composite Membrane with Binary Structures for Effective Air Filtration.

    Science.gov (United States)

    Zhang, Shichao; Liu, Hui; Yin, Xia; Yu, Jianyong; Ding, Bin

    2016-03-01

    Airborne particle filtration proposed for fibers requires their assembly into porous structures with small pore size and low packing density. The ability to maintain structural stability upon deformation stress in service is essential to ensure a highly porous packing material that functions reliably; however, it has proven extremely challenging. Here, we report a strategy to create anti-deformed poly(ethylene oxide)@polyacrylonitrile/polysulfone (PEO@PAN/PSU) composite membranes with binary structures for effective air filtration by combining multijet electrospinning and physical bonding process. Our approach allows the ambigenous fiber framework including thin PAN nanofibers and fluffy PSU microfibers, through which run interpenetrating PEO bonding structures, to assemble into stable filtration medium with tunable pore size and packing density by facilely optimizing the bimodal fiber construction and benefiting from the PEO inspiration. With the integrated features of small pore size, high porosity, and robust mechanical properties (8.2 MPa), the resultant composite membrane exhibits high filtration efficiency of 99.992%, low pressure drop of 95 Pa, and desirable quality factor of 0.1 Pa(-1); more significantly, it successfully gets rid of the potential safety hazards caused by unexpected structural collapsing under service stress. The synthesis of PEO@PAN/PSU medium would not only make it a promising candidate for PM2.5 governance but also provide a versatile strategy to design and develop stable porous membranes for various applications.

  20. Structure and Function of the Membrane Deformation AAA ATPase Vps4

    Science.gov (United States)

    Hill, Christopher P.; Babst, Markus

    2011-01-01

    The ATPase Vps4 belongs to the type-I AAA family of proteins. Vps4 functions together with a group of proteins referred to as ESCRTs in membrane deformation and fission events. These cellular functions include vesicle formation at the endosome, cytokinesis and viral budding. The highly dynamic quaternary structure of Vps4 and its interactions with a network of regulators and co-factors have made the analysis of this ATPase challenging. Nevertheless, recent advances in the understanding of the cell biology of Vps4 together with structural information and in vitro studies are guiding mechanistic models of this ATPase. PMID:21925211

  1. Dynamics of a bilayer membrane coupled to a two-dimensional cytoskeleton: Scale transfers of membrane deformations

    Science.gov (United States)

    Okamoto, Ryuichi; Komura, Shigeyuki; Fournier, Jean-Baptiste

    2017-07-01

    We theoretically investigate the dynamics of a floating lipid bilayer membrane coupled with a two-dimensional cytoskeleton network, taking into account explicitly the intermonolayer friction, the discrete lattice structure of the cytoskeleton, and its prestress. The lattice structure breaks lateral continuous translational symmetry and couples Fourier modes with different wave vectors. It is shown that within a short time interval a long-wavelength deformation excites a collection of modes with wavelengths shorter than the lattice spacing. These modes relax slowly with a common renormalized rate originating from the long-wavelength mode. As a result, and because of the prestress, the slowest relaxation is governed by the intermonolayer friction. Conversely, and most interestingly, forces applied at the scale of the cytoskeleton for a sufficiently long time can cooperatively excite large-scale modes.

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

    Science.gov (United States)

    Ozkaleli, Merve; Erdem, Ayca

    2018-02-27

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

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

    Directory of Open Access Journals (Sweden)

    Merve Ozkaleli

    2018-02-01

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

  4. Monocytic cells become less compressible but more deformable upon activation.

    Directory of Open Access Journals (Sweden)

    Agnese Ravetto

    Full Text Available Monocytes play a significant role in the development of atherosclerosis. During the process of inflammation, circulating monocytes become activated in the blood stream. The consequent interactions of the activated monocytes with the blood flow and endothelial cells result in reorganization of cytoskeletal proteins, in particular of the microfilament structure, and concomitant changes in cell shape and mechanical behavior. Here we investigate the full elastic behavior of activated monocytes in relation to their cytoskeletal structure to obtain a better understanding of cell behavior during the progression of inflammatory diseases such as atherosclerosis.The recently developed Capillary Micromechanics technique, based on exposing a cell to a pressure difference in a tapered glass microcapillary, was used to measure the deformation of activated and non-activated monocytic cells. Monitoring the elastic response of individual cells up to large deformations allowed us to obtain both the compressive and the shear modulus of a cell from a single experiment. Activation by inflammatory chemokines affected the cytoskeletal organization and increased the elastic compressive modulus of monocytes with 73-340%, while their resistance to shape deformation decreased, as indicated by a 25-88% drop in the cell's shear modulus. This decrease in deformability is particularly pronounced at high strains, such as those that occur during diapedesis through the vascular wall.Overall, monocytic cells become less compressible but more deformable upon activation. This change in mechanical response under different modes of deformation could be important in understanding the interplay between the mechanics and function of these cells. In addition, our data are of direct relevance for computational modeling and analysis of the distinct monocytic behavior in the circulation and the extravascular space. Lastly, an understanding of the changes of monocyte mechanical properties

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

    Science.gov (United States)

    Li, Zhen-Lu

    2018-03-01

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

  6. Investigating the effects of membrane deformability on artificial capsule adhesion to the functionalized surface.

    Science.gov (United States)

    Balsara, Hiren D; Banton, Rohan J; Eggleton, Charles D

    2016-10-01

    Understanding, manipulating and controlling cellular adhesion processes can be critical in developing biomedical technologies. Adhesive mechanisms can be used to the target, pattern and separate cells such as leukocytes from whole blood for biomedical applications. The deformability response of the cell directly affects the rolling and adhesion behavior under viscous linear shear flow conditions. To that end, the primary objective of the present study was to investigate numerically the influence of capsule membrane's nonlinear material behavior (i.e. elastic-plastic to strain hardening) on the rolling and adhesion behavior of representative artificial capsules. Specifically, spherical capsules with radius of [Formula: see text] were represented using an elastic membrane governed by a Mooney-Rivlin strain energy functions. The surfaces of the capsules were coated with P-selectin glycoprotein-ligand-1 to initiate binding interaction with P-selectin-coated planar surface with density of [Formula: see text] under linear shear flow varying from 100 to [Formula: see text]. The numerical model is based on the Immersed Boundary Method for rolling of deformable capsule in shear flow coupled with Monte Carlo simulation for receptor/ligand interaction modeled using Bell model. The results reveal that the mechanical properties of the capsule play an important role in the rolling behavior and the binding kinetics between the capsule contact surface and the substrate. The rolling behavior of the strain hardening capsules is relatively smoother and slower compared to the elastic-plastic capsules. The strain hardening capsules exhibits higher contact area at any given shear rate compared to elastic-plastic capsules. The increase in contact area leads to decrease in rolling velocity. The capsule contact surface is not in complete contact with the substrate because of thin lubrication film that is trapped between the capsule and substrate. This creates a concave shape on the bottom

  7. Deformation of Two-Dimensional Nonuniform-Membrane Red Blood Cells Simulated by a Lattice Boltzmann Model

    International Nuclear Information System (INIS)

    Hua-Bing, Li; Li, Jin; Bing, Qiu

    2008-01-01

    To study two-dimensional red blood cells deforming in a shear Bow with the membrane nonuniform on the rigidity and mass, the membrane is discretized into equilength segments. The fluid inside and outside the red blood cell is simulated by the D2Q9 lattice Boltzmann model and the hydrodynamic forces exerted on the membrane from the inner and outer of the red blood cell are calculated by a stress-integration method. Through the global deviation from the curvature of uniform-membrane, we find that when the membrane is nonuniform on the rigidity, the deviation first decreases with the time increases and implies that the terminal profile of the red blood cell is static. To a red blood cell with the mass nonuniform on the membrane, the deviation becomes more large, and the mass distribution affects the profile of the two sides of the flattened red blood cell in a shear flow. (fundamental areas of phenomenology(including applications))

  8. Characterization of acoustically induced deformations of human tympanic membranes by digital holography and shearography

    Science.gov (United States)

    Flores-Moreno, J. M.; Furlong, Cosme; Cheng, Jeffrey T.; Rosowski, John J.; Merchant, S. N.

    2011-08-01

    Recently, we introduced a Digital Optoelectronic Holographic System (DOEHS) for measurement of acoustically induced deformations of the human tympanic membrane (TM) in order to study and diagnose pathologic conditions of the middle-ear. The DOEHS consists of laser-delivery illumination (IS), optical head (OH), image-processing computer (IP), and positioning arm (PS) subsystems. Holographic information is recorded by a CCD and numerically reconstructed by Fresnel approximation. Our holographic otoscope system is currently deployed in a clinic and is packaged in a custom design. Since digital holography is a high sensitivity measurement technique and the interfering light waves travel along different paths, it makes measurements acquired by DOEHS susceptible to external vibrations. In order to avoid this susceptibility, we are testing a shearography setup as OH. Shearography presents same advantages as holographic interferometry, but it is less susceptible to vibration and external noise, which is a characteristic needed for the use of our techniques in a clinical environment. In this paper we present work in progress in our development of a shearography technique based on a Mach-Zehnder configuration as OH and demonstrate its application by quantifying vibrations modes in thin membranes, including human TM. Results are compared with those obtained with DOEHS.

  9. Amoeboid swimming: a generic self-propulsion of cells in fluids by means of membrane deformations.

    Science.gov (United States)

    Farutin, Alexander; Rafaï, Salima; Dysthe, Dag Kristian; Duperray, Alain; Peyla, Philippe; Misbah, Chaouqi

    2013-11-27

    Microorganisms, such as bacteria, algae, or spermatozoa, are able to propel themselves forward thanks to flagella or cilia activity. By contrast, other organisms employ pronounced changes of the membrane shape to achieve propulsion, a prototypical example being the Eutreptiella gymnastica. Cells of the immune system as well as dictyostelium amoebas, traditionally believed to crawl on a substratum, can also swim in a similar way. We develop a model for these organisms: the swimmer is mimicked by a closed incompressible membrane with force density distribution (with zero total force and torque). It is shown that fast propulsion can be achieved with adequate shape adaptations. This swimming is found to consist of an entangled pusher-puller state. The autopropulsion distance over one cycle is a universal linear function of a simple geometrical dimensionless quantity A/V(2/3) (V and A are the cell volume and its membrane area). This study captures the peculiar motion of Eutreptiella gymnastica with simple force distribution.

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

    Science.gov (United States)

    Boyko, Evgeniy; Gat, Amir; Bercovici, Moran

    2017-11-01

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

  11. Root hair deformation activity of nodulation factor and their fate on Vicia sativa.

    NARCIS (Netherlands)

    Heidstra, R.; Geurts, R.; Franssen, H.; Spaink, H.P.; Kammen, van A.; Bisseling, T.

    1994-01-01

    We used a semiquantitative root hair deformation assay for Vicia sativa (vetch) to study the activity of Rhizobium leguminosarum bv viciae nodulation (Nod) factors. Five to 10 min of Nod factor-root interaction appears to be sufficient to induce root hair deformation. The first deformation is

  12. Active processes make mixed lipid membranes either flat or crumpled

    Science.gov (United States)

    Banerjee, Tirthankar; Basu, Abhik

    2018-01-01

    Whether live cell membranes show miscibility phase transitions (MPTs), and if so, how they fluctuate near the transitions remain outstanding unresolved issues in physics and biology alike. Motivated by these questions we construct a generic hydrodynamic theory for lipid membranes that are active, due for instance, to the molecular motors in the surrounding cytoskeleton, or active protein components in the membrane itself. We use this to uncover a direct correspondence between membrane fluctuations and MPTs. Several testable predictions are made: (i) generic active stiffening with orientational long range order (flat membrane) or softening with crumpling of the membrane, controlled by the active tension and (ii) for mixed lipid membranes, capturing the nature of putative MPTs by measuring the membrane conformation fluctuations. Possibilities of both first and second order MPTs in mixed active membranes are argued for. Near second order MPTs, active stiffening (softening) manifests as a super-stiff (super-soft) membrane. Our predictions are testable in a variety of in vitro systems, e.g. live cytoskeletal extracts deposited on liposomes and lipid membranes containing active proteins embedded in a passive fluid.

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

    Science.gov (United States)

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

    2017-12-01

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

  14. Chemical activity of cholesterol in membranes.

    Science.gov (United States)

    Radhakrishnan, A; McConnell, H M

    2000-07-18

    Measurements are reported for the rate constants for the release of cholesterol (and dihydrocholesterol) to beta-cyclodextrin from mixtures with phospholipids in homogeneous monolayers at constant pressure at the air-water interface. In each mixture, it is found that the release rate shows a sharp decrease as the cholesterol concentration in the monolayer decreases through a composition corresponding to the stoichiometry of a cholesterol-phospholipid complex. The stoichiometry of the complex was established previously by the position of a sharp cusp in the thermodynamic phase diagram of each mixture and also by a minimum in average molecular area versus composition measurements. A theoretical model used earlier to account for the phase diagrams predicts the chemical potential and chemical activity of cholesterol in these mixtures. The calculated chemical activity also shows a sharp change at the complex stoichiometry in homogeneous monolayers. The similarities in change of observed release rate and calculated chemical activity are expected from reaction rate theory where the release rate is proportional to the cholesterol chemical activity. The chemical activity of cholesterol as determined by complex formation between some phospholipids and cholesterol in the plasma membrane of cells may serve a regulatory function with respect to intracellular cholesterol transport and biosynthesis.

  15. Electrospun nanofiber membranes for electrically activated shape memory nanocomposites

    Science.gov (United States)

    Zhang, Fenghua; Zhang, Zhichun; Liu, Yanju; Leng, Jinsong

    2014-06-01

    A novel shape memory nanocomposite system, consisting of a thermoplastic Nafion polymer and ultrathin electrospun polyacrylonitrile (PAN)-based carbonization nanofiber membranes, is successfully synthesized. PAN-based carbonization nanofiber networks that offer responses to deformations are considered to be an excellent actuation source. Significant improvement in the electrical conductivity of carbon nanofiber membranes is found by adjusting the applied voltage power in the electrospinning PAN process varying from 7.85 to 12.30 S cm-1. The porous structure of the carbon nanofiber membranes provides a large specific surface area and interfacial contact area when combined with the polymer matrix. Shape memory Nafion nanocomposites filled with interpenetrating non-woven electrospun PAN carbonization membranes can be actuated by applying 14 V electrical voltage within 5 s. The results, as demonstrated through morphology, electrical and thermal measurements and a shape recovery test, suggest a valuable route to producing soft nanocomposites.

  16. Following-up changes in red blood cell deformability and membrane stability in the presence of PTFE graft implanted into the femoral artery in a canine model

    Science.gov (United States)

    Toth, Csaba; Kiss, Ferenc; Klarik, Zoltan; Gergely, Eszter; Toth, Eniko; Peto, Katalin; Vanyolos, Erzsebet; Miko, Iren; Nemeth, Norbert

    2014-05-01

    It is known that a moderate mechanical stress can even improve the red blood cells' (RBC) micro-rheological characteristics, however, a more significant stress causes deterioration in the deformability. In this study, we aimed to investigate the effect of the presence of artificial graft on the RBC deformability and membrane stability in beagles. In the Control group only anesthesia was induced and in the postoperative (p.o.) period blood samplings were carried out. In the Grafted group under general anesthesia, the left femoral artery was isolated, from which a 3.5 cm segment was resected and a PTFE graft (O.D.: 3 mm) of equal in length was implanted into the gap. On the 1st, 3rd, 5th, 7th and 14th p.o. days blood was collected the cephalic veins and RBC deformability was determined ektacytometry (LoRRca MaxSis Osmoscan). Membrane stability test consisted of two deformability measurements before and after the cells were being exposed to mechanical stress (60 or 100 Pa for 300 seconds). Compared to the Control group and the baseline values the red blood cell deformability showed significant deterioration on the 3rd, 5th and mainly on the 7th postoperative day after the graft implantation. The membrane stability of erythrocyte revealed marked inter-group difference on the 3rd, 5th and 7th day: in the Grafted group the deformability decreased and during the membrane stability test smaller difference was observed between the states before and after shearing. We concluded that the presence of a PTFE graft in the femoral artery may cause changes in RBC deformability in the first p.o. week. RBC membrane stability investigation shows a lower elongation index profile for the grafted group and a narrowed alteration in the deformability curves due to mechanical stress.

  17. Abba promotes PDGF-mediated membrane ruffling through activation of the small GTPase Rac1.

    Science.gov (United States)

    Zheng, Datong; Niu, Shuqiong; Yu, Dan; Zhan, Xiaoguo H; Zeng, Xianchun; Cui, Bota; Chen, Yanping; Yoon, Jennifer; Martin, Stuart S; Lu, Xiang; Zhan, Xi

    2010-10-29

    Abba is a member of the I-BAR-domain protein family that is characterized by a convex-shaped membrane-binding motif. Overexpression of GFP-tagged Abba in murine fibroblasts potentiated PDGF-mediated formation of membrane ruffles and lamellipodia. Immunofluorescent microscopy and pull-down analysis revealed that GFP-Abba colocalized with an active form of Rac1 in the membrane ruffles and enhanced the Rac GTPase activity in response to PDGF stimulation. Further immunoprecipitation assays demonstrated that GFP-Abba bound to both wild-type and constitutively active Rac1 and that the binding to either of the Rac1 forms was significantly enhanced upon PDGF stimulation. On the other hand, an Abba mutant deficient in Rac1 binding failed to promote membrane ruffling and Rac1 activation in response to PDGF. However, the cells overexpressing a truncated mutant carrying the I-BAR domain alone displayed numerous filopodia-like microspikes in a manner independent of growth factors. Also, the Rac-binding activity of the mutant was not affected by PDGF treatment. Our data indicates that the interaction between full-length Abba and Rac1 is implicated in membrane deformation and subjected to a growth factor-mediated regulation through the C-terminal sequence. Copyright © 2010 Elsevier Inc. All rights reserved.

  18. Numerical study of bimetallic actuated micro-membrane with large deformations

    International Nuclear Information System (INIS)

    Lefevre, R; Salette, A; Agraffeil, C; Guillen, J; Morfouli, P; Montès, L; Déhan, C

    2013-01-01

    This paper presents a new method to better predict bimetallic thermally actuated membrane behavior in micro-electro-mechanical systems based on finite element method simulations. Unlike previous approaches, this new method includes heat transfer analysis to evaluate the temperature distribution under definite power actuation. Large deflection behavior (larger than one half of the membrane thickness) is predicted from the evaluated temperature distribution and additional applied pressure. Ultra-thin membranes for high displacements have been fabricated using an IC-compatible process flow. Their thermal and mechanical characteristics are compared with simulation predictions. The results under pressure show a lower than 4% difference between experiments and theoretical predictions. (paper)

  19. Highly Reflecting, Broadband Deformable Membrane Mirror for Wavefront Control Applications, Phase I

    Data.gov (United States)

    National Aeronautics and Space Administration — This Phase I STTR project will develop a highly reflecting, broadband, radiation resistant, low-stress and lightweight, membrane integrated into an electrostatically...

  20. Large deformation contact mechanics of a pressurized long rectangular membrane. II. Adhesive contact

    Science.gov (United States)

    Srivastava, Abhishek; Hui, Chung-Yuen

    2013-01-01

    In part I of this work, we presented a theory for adhesionless contact of a pressurized neo-Hookean plane-strain membrane to a rigid substrate. Here, we extend our theory to include adhesion using a fracture mechanics approach. This theory is used to study contact hysteresis commonly observed in experiments. Detailed analysis is carried out to highlight the differences between frictionless and no-slip contact. Membrane detachment is found to be strongly dependent on adhesion: for low adhesion, the membrane ‘pinches-off’, whereas for large adhesions, it detaches unstably at finite contact (‘pull-off’). Expressions are derived for the critical adhesion needed for pinch-off to pull-off transition. Above a threshold adhesion, the membrane exhibits bistability, two stable states at zero applied pressure. The condition for bistability for both frictionless and no-slip boundary conditions is obtained explicitly. PMID:24353472

  1. Measurements of Active Tectonic Deformation on the Guerrero Coast, Mexico

    Science.gov (United States)

    Ramirez, T.; Cundy, A.; Carranza-Edwards, A.; Morales, E.; Kostoglodov, V.; Urrutia-Fucugauchi, J.

    2004-12-01

    The study of tectonic deformation rates using displaced shoreline features is relatively well-established, and has provided much useful information on seismic hazard. Such studies have frequently been complemented by analysis of the coastal sedimentary record, where past marine to terrestrial environmental changes (and vice versa) may be recorded by clear changes in stratigraphy. Studies of this type are particularly valuable for tectonically-active areas where the preservation of former shoreline features is poor, or where long-term subsidence has resulted in their erosion, drowning or burial. The specific objective of this study is to derive rates of tectonic deformation from geomorphic and stratigraphic studies of the Guerrero coastal area, and to examine the feasibility of this stratigraphic approach in the coastal lagoons of the Mexican Pacific coast, in the Guerrero gap. The Guerrero gap coastal area, where a major earthquake is expected to occur, parallels the Cocos plate subduction zone. Here convergence rates vary from 5.2 cm/yr to 5.8 cm/yr. The Guerrero gap has experienced several historical earthquakes, notably the 1911 (7.8 Ms). However, no large magnitude events since the 1911 earthquake and only a few Ms~6 events have occurred near the Guerrero gap edges. It is expected that a major interplate earthquake of estimated magnitude Mw=8.1 to 8.4 has a high probability to occur. Landforms within the Guerrero gap indicate that the coast is subsiding. A series of key indicators such as elongated islands reminiscent of ancient barriers, submerged barriers island, extensive marshy environments, increased depths in the lagoons, and submerged anthropogenic features (shell mounds), among others, suggest active tectonic subsidence of the coast. In contrast, the adjacent northwest area off the Guerrero gap exhibits landforms characteristic of tectonic uplift (marine terraces and uplifted beach ridges), indicating a different seismo-tectonic regime northwest of the

  2. Membrane Stabilizing Activity And Phytochemistry Of Hibiscus rosa ...

    African Journals Online (AJOL)

    The human erythrocyte membrane stabilizing activity of saline extract of Hibiscus rosa-sinensis leaves was investigated as part of efforts at validating its use as anti-arthritic and anti-inflammatory agent. The results of the membrane stabilizing activity of the extract, when compared to two non-steroidal anti-inflammatory drugs ...

  3. Development of active-transport membrane devices

    Energy Technology Data Exchange (ETDEWEB)

    Laciak, D.V.

    1994-07-01

    This report introduces the concept of Air Products` AT membranes for the separation of NH{sub 3} and CO{sub 2} from process gas streams and presents results from the first year fabrication concept development studies.

  4. Active membrane having uniform physico-chemically functionalized ion channels

    Science.gov (United States)

    Gerald, II, Rex E; Ruscic, Katarina J; Sears, Devin N; Smith, Luis J; Klingler, Robert J; Rathke, Jerome W

    2012-09-24

    The present invention relates to a physicochemically-active porous membrane for electrochemical cells that purports dual functions: an electronic insulator (separator) and a unidirectional ion-transporter (electrolyte). The electrochemical cell membrane is activated for the transport of ions by contiguous ion coordination sites on the interior two-dimensional surfaces of the trans-membrane unidirectional pores. One dimension of the pore surface has a macroscopic length (1 nm-1000 .mu.m) and is directed parallel to the direction of an electric field, which is produced between the cathode and the anode electrodes of an electrochemical cell. The membrane material is designed to have physicochemical interaction with ions. Control of the extent of the interactions between the ions and the interior pore walls of the membrane and other materials, chemicals, or structures contained within the pores provides adjustability of the ionic conductivity of the membrane.

  5. Effects of deformability and thermal motion of lipid membrane on electroporation: By molecular dynamics simulations

    KAUST Repository

    Sun, Sheng

    2011-01-01

    Effects of mechanical properties and thermal motion of POPE lipid membrane on electroporation were studied by molecular dynamics simulations. Among simulations in which specific atoms of lipids were artificially constrained at their equilibrium positions using a spring with force constant of 2.0kcal/(molÅ2) in the external electric field of 1.4kcal/(molÅe), only constraint on lateral motions of lipid tails prohibited electroporation while non-tail parts had little effects. When force constant decreased to 0.2kcal/(molÅ2) in the position constraints on lipid tails in the external electric field of 2.0kcal/(molÅe), water molecules began to enter the membrane. Position constraints of lipid tails allow water to penetrate from both sides of membrane. Thermal motion of lipids can induce initial defects in the hydrophobic core of membrane, which are favorable nucleation sites for electroporation. Simulations at different temperatures revealed that as the temperature increases, the time taken to the initial pore formation will decrease. © 2010 Elsevier Inc.

  6. Turing Incompleteness of Asynchronous P Systems with Active Membranes

    OpenAIRE

    Leporati, Alberto; Manzoni, Luca; Porreca, Antonio E.

    2013-01-01

    We prove that asynchronous P systems with active membranes without divi- sion rules can be simulated by place/transition Petri nets, and hence are computationally weaker than Turing machines. This result holds even if the synchronisation mechanisms provided by electrical charges and membrane dissolution are exploited.

  7. Large deformation and mechanics of flexible isotropic membrane ballooning in three dimensions by differential quadrature method

    International Nuclear Information System (INIS)

    Mozaffari, M.; Atai, A. A.; Mostafa, N.

    2009-01-01

    This paper presents a computationally efficient and accurate new methodology in the differential quadrature analysis of structural mechanics for flexible membranes ballooning in three dimensions under a negative air pressure differential. The differential quadrature method is employed to discretize the resulting equations in the axial direction as well as for the solution procedure. The weighting coefficients employed are not exclusive, and any accurate and efficient method such as the generalized differential quadrature method may be used to produce the methods weighting coefficients. A second-order paraboloid of revolution is assumed to describe the ballooning shape. This study asserts the accuracy, convergency, and efficiency of the methodology by solving some typical stability, straining analysis membrane problems, and comparing the results with those of the exact solutions and/or those of physical tests

  8. Large deformation and mechanics of flexible isotropic membrane ballooning in three dimensions by differential quadrature method

    Energy Technology Data Exchange (ETDEWEB)

    Mozaffari, M.; Atai, A. A.; Mostafa, N. [Islamic Azad University, Karaj (Iran, Islamic Republic of)

    2009-11-15

    This paper presents a computationally efficient and accurate new methodology in the differential quadrature analysis of structural mechanics for flexible membranes ballooning in three dimensions under a negative air pressure differential. The differential quadrature method is employed to discretize the resulting equations in the axial direction as well as for the solution procedure. The weighting coefficients employed are not exclusive, and any accurate and efficient method such as the generalized differential quadrature method may be used to produce the methods weighting coefficients. A second-order paraboloid of revolution is assumed to describe the ballooning shape. This study asserts the accuracy, convergency, and efficiency of the methodology by solving some typical stability, straining analysis membrane problems, and comparing the results with those of the exact solutions and/or those of physical tests

  9. Modeling crustal deformation near active faults and volcanic centers: a catalog of deformation models and modeling approaches

    Science.gov (United States)

    Battaglia, Maurizio; ,; Peter, F.; Murray, Jessica R.

    2013-01-01

    This manual provides the physical and mathematical concepts for selected models used to interpret deformation measurements near active faults and volcanic centers. The emphasis is on analytical models of deformation that can be compared with data from the Global Positioning System (GPS) receivers, Interferometric synthetic aperture radar (InSAR), leveling surveys, tiltmeters and strainmeters. Source models include pressurized spherical, ellipsoidal, and horizontal penny-shaped geometries in an elastic, homogeneous, flat half-space. Vertical dikes and faults are described following the mathematical notation for rectangular dislocations in an elastic, homogeneous, flat half-space. All the analytical expressions were verified against numerical models developed by use of COMSOL Multyphics, a Finite Element Analysis software (http://www.comsol.com). In this way, typographical errors present were identified and corrected. Matlab scripts are also provided to facilitate the application of these models.

  10. An Extended Membrane System with Active Membranes to Solve Automatic Fuzzy Clustering Problems.

    Science.gov (United States)

    Peng, Hong; Wang, Jun; Shi, Peng; Pérez-Jiménez, Mario J; Riscos-Núñez, Agustín

    2016-05-01

    This paper focuses on automatic fuzzy clustering problem and proposes a novel automatic fuzzy clustering method that employs an extended membrane system with active membranes that has been designed as its computing framework. The extended membrane system has a dynamic membrane structure; since membranes can evolve, it is particularly suitable for processing the automatic fuzzy clustering problem. A modification of a differential evolution (DE) mechanism was developed as evolution rules for objects according to membrane structure and object communication mechanisms. Under the control of both the object's evolution-communication mechanism and the membrane evolution mechanism, the extended membrane system can effectively determine the most appropriate number of clusters as well as the corresponding optimal cluster centers. The proposed method was evaluated over 13 benchmark problems and was compared with four state-of-the-art automatic clustering methods, two recently developed clustering methods and six classification techniques. The comparison results demonstrate the superiority of the proposed method in terms of effectiveness and robustness.

  11. Silver-enhanced block copolymer membranes with biocidal activity

    KAUST Repository

    Madhavan, Poornima

    2014-11-12

    Silver nanoparticles were deposited on the surface and pore walls of block copolymer membranes with highly ordered pore structure. Pyridine blocks constitute the pore surfaces, complexing silver ions and promoting a homogeneous distribution. Nanoparticles were then formed by reduction with sodium borohydride. The morphology varied with the preparation conditions (pH and silver ion concentration), as confirmed by field emission scanning and transmission electron microscopy. Silver has a strong biocide activity, which for membranes can bring the advantage of minimizing the growth of bacteria and formation of biofilm. The membranes with nanoparticles prepared under different pH values and ion concentrations were incubated with Pseudomonas aeruginosa and compared with the control. The strongest biocidal activity was achieved with membranes containing membranes prepared under pH 9. Under these conditions, the best distribution with small particle size was observed by microscopy.

  12. Characterization of the activation of small GTPases by their GEFs on membranes using artificial membrane tethering.

    Science.gov (United States)

    Peurois, François; Veyron, Simon; Ferrandez, Yann; Ladid, Ilham; Benabdi, Sarah; Zeghouf, Mahel; Peyroche, Gérald; Cherfils, Jacqueline

    2017-03-23

    Active, GTP-bound small GTPases need to be attached to membranes by post-translational lipid modifications in order to process and propagate information in cells. However, generating and manipulating lipidated GTPases has remained difficult, which has limited our quantitative understanding of their activation by guanine nucleotide exchange factors (GEFs) and their termination by GTPase-activating proteins. Here, we replaced the lipid modification by a histidine tag in 11 full-length, human small GTPases belonging to the Arf, Rho and Rab families, which allowed to tether them to nickel-lipid-containing membranes and characterize the kinetics of their activation by GEFs. Remarkably, this strategy uncovered large effects of membranes on the efficiency and/or specificity in all systems studied. Notably, it recapitulated the release of autoinhibition of Arf1, Arf3, Arf4, Arf5 and Arf6 GTPases by membranes and revealed that all isoforms are efficiently activated by two GEFs with different regulatory regimes, ARNO and Brag2. It demonstrated that membranes stimulate the GEF activity of Trio toward RhoG by ∼30 fold and Rac1 by ∼10 fold, and uncovered a previously unknown broader specificity toward RhoA and Cdc42 that was undetectable in solution. Finally, it demonstrated that the exceptional affinity of the bacterial RabGEF DrrA for the phosphoinositide PI(4)P delimits the activation of Rab1 to the immediate vicinity of the membrane-bound GEF. Our study thus validates the histidine-tag strategy as a potent and simple means to mimic small GTPase lipidation, which opens a variety of applications to uncover regulations brought about by membranes. © 2017 The Author(s); published by Portland Press Limited on behalf of the Biochemical Society.

  13. Guanidination of notexin alters its membrane-damaging activity in ...

    Indian Academy of Sciences (India)

    MADHU

    EYSM/cholesterol vesicles were conducted. EYSM notably reduced the membrane-damaging activity of notexin against EYPC vesicles, but had an insignificant influence on that of Gu-notexin. Unlike the effects noted with notexin, inactivation of ...

  14. The Research of Phase Retrieval Holography Method Based on the Active Deformation of the Active Reflector Surface

    Science.gov (United States)

    Wang, Z. Q.; Chen, M. Z.; Pei, X.; Wang, J.

    2017-09-01

    The surface accuracy of a large reflector radio telescope is one of the important factors influencing the performance of the antenna. The effects of panel processing, installation, as well as gravity, temperature, and wind load, will greatly limit the observation efficiency of the antenna. Focused on the technology of active surface which is more accurately controllable than the minor reflector surface of six-ploe, the continuous distribution of active deformation phase factor described by Zernike polynomials is adopted for the first time. Only getting the far field amplitude through adjusting the active surface, the surface error can be detected. By building the models of numerical simulation, the retrieval error of arbitrary surface deformation is calculated, and the retrieval results of surface deformation in a variety of continuous active surface deformation is also studied. It is indicated that this method can stably and accurately detect surface deformation, and can also improve the efficiency of radio telescope observations effectively.

  15. Lipase immobilized catalytically active membrane for synthesis of lauryl stearate in a pervaporation membrane reactor.

    Science.gov (United States)

    Zhang, Weidong; Qing, Weihua; Ren, Zhongqi; Li, Wei; Chen, Jiangrong

    2014-11-01

    A composite catalytically active membrane immobilized with Candida rugosa lipase has been prepared by immersion phase inversion technique for enzymatic synthesis of lauryl stearate in a pervaporation membrane reactor. SEM images showed that a "sandwich-like" membrane structure with a porous lipase-PVA catalytic layer uniformly coated on a polyvinyl alcohol (PVA)/polyethersulfone (PES) bilayer was obtained. Optimum conditions for lipase immobilization in the catalytic layer were determined. The membrane was proved to exhibit superior thermal stability, pH stability and reusability than free lipase under similar conditions. In the case of pervaporation coupled synthesis of lauryl stearate, benefited from in-situ water removal by the membrane, a conversion enhancement of approximately 40% was achieved in comparison to the equilibrium conversion obtained in batch reactors. In addition to conversion enhancement, it was also found that excess water removal by the catalytically active membrane appears to improve activity of the lipase immobilized. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. Redistribution of Cholesterol in Model Lipid Membranes in Response to the Membrane-Active Peptide Alamethicin

    Science.gov (United States)

    Heller, William; Qian, Shuo

    2013-03-01

    The cellular membrane is a heterogeneous, dynamic mixture of molecules and macromolecules that self-assemble into a tightly-regulated functional unit that provides a semipermeable barrier between the cell and its environment. Among the many compositional differences between mammalian and bacterial cell membranes that impact its physical properties, one key difference is cholesterol content, which is more prevalent in mammals. Cholesterol is an amphiphile that associates with membranes and serves to maintain its fluidity and permeability. Membrane-active peptides, such as the alpha-helical peptide alamethicin, interact with membranes in a concentration- and composition-dependent manner to form transmembrane pores that are responsible for the lytic action of the peptide. Through the use of small-angle neutron scattering and deuterium labeling, it was possible to observe a redistribution of the lipid and cholesterol in unilamellar vesicles in response to the presence of alamethicin at a peptide-to-lipid ratio of 1/200. The results demonstrate that the membrane remodeling powers of alamethicin reach beyond the membrane thinning effect to altering the localization of specific components in the bilayer, complementing the accepted two-state mechanism of pore formation. Research was supported by U. S. DOE-OBER (CSMB; FWP ERKP291) and the U. S. DOE-BES Scientific User Facilities Division (ORNL's SNS and HFIR).

  17. Ovalbumin with Glycated Carboxyl Groups Shows Membrane-Damaging Activity

    Directory of Open Access Journals (Sweden)

    Ching-Chia Tang

    2017-02-01

    Full Text Available The aim of the present study was to investigate whether glycated ovalbumin (OVA showed novel activity at the lipid-water interface. Mannosylated OVA (Man-OVA was prepared by modification of the carboxyl groups with p-aminophenyl α-dextro (d-mannopyranoside. An increase in the number of modified carboxyl groups increased the membrane-damaging activity of Man-OVA on cell membrane-mimicking vesicles, whereas OVA did not induce membrane permeability in the tested phospholipid vesicles. The glycation of carboxyl groups caused a notable change in the gross conformation of OVA. Moreover, owing to their spatial positions, the Trp residues in Man-OVA were more exposed, unlike those in OVA. Fluorescence quenching studies suggested that the Trp residues in Man-OVA were located on the interface binds with the lipid vesicles, and their microenvironment was abundant in positively charged residues. Although OVA and Man-OVA showed a similar binding affinity for lipid vesicles, the lipid-interacting feature of Man-OVA was distinct from that of OVA. Chemical modification studies revealed that Lys and Arg residues, but not Trp residues, played a crucial role in the membrane-damaging activity of Man-OVA. Taken together, our data suggest that glycation of carboxyl groups causes changes in the structural properties and membrane-interacting features of OVA, generating OVA with membrane-perturbing activities at the lipid-water interface.

  18. Deformation of a membrane in a pulsatile flow: implications in heart valve design

    Science.gov (United States)

    Hernandez, C.; Guzman, J. E. V.; Zenit, R.

    2011-11-01

    Current designs of heart valves prosthetics have serious disadvantages and health issues for patients who use them. For this reason, a new design that combines durability (mechanical valves) and biocompatibility (biological valves) has to be conceived. Natural valves have very complex geometry because their leaflets have two principal curvatures, one imposed by the holding ring and a second one imposed by the bending of the closing arrangement. The objective of this research is to study the effects of both curvatures on the performance of a leaflet. It is well known that the increase of the curvature results in a larger stiffness, which, in turn, reduces the deflection of a leaflet. We conducted a study to determine the effect of changing the curvature (in two directions) of a flexible membrane when exposed to a steady and pulsatile flows. A study of the flow field that results from this interaction is also conducted by PIV measurements. Preliminary results of the leaflet deflection for many stiffnesses, curvatures and flow conditions will be presented and discussed.

  19. Synthetic Molecular Evolution of Membrane-Active Peptides

    Science.gov (United States)

    Wimley, William

    The physical chemistry of membrane partitioning largely determines the function of membrane active peptides. Membrane-active peptides have potential utility in many areas, including in the cellular delivery of polar compounds, cancer therapy, biosensor design, and in antibacterial, antiviral and antifungal therapies. Yet, despite decades of research on thousands of known examples, useful sequence-structure-function relationships are essentially unknown. Because peptide-membrane interactions within the highly fluid bilayer are dynamic and heterogeneous, accounts of mechanism are necessarily vague and descriptive, and have little predictive power. This creates a significant roadblock to advances in the field. We are bypassing that roadblock with synthetic molecular evolution: iterative peptide library design and orthogonal high-throughput screening. We start with template sequences that have at least some useful activity, and create small, focused libraries using structural and biophysical principles to design the sequence space around the template. Orthogonal high-throughput screening is used to identify gain-of-function peptides by simultaneously selecting for several different properties (e.g. solubility, activity and toxicity). Multiple generations of iterative library design and screening have enabled the identification of membrane-active sequences with heretofore unknown properties, including clinically relevant, broad-spectrum activity against drug-resistant bacteria and enveloped viruses as well as pH-triggered macromolecular poration.

  20. Activity and distribution of urease following microencapsulation within polyamide membranes.

    Science.gov (United States)

    Monshipouri, M; Neufeld, R J

    1991-04-01

    Urease was microencapsulated by forming a semipermeable polyamide membrane around aqueous microdroplets (266 microns mean diameter) containing the soluble enzyme. The yield of the interfacial polymerization technique, determined spectrophotometrically, was 83% of the original enzyme on a mass basis, resulting in a final intracapsular urease concentration of 62.3 mg ml-1 or 0.1 mM. Similar absorption spectra of broken and intact microcapsules suggested that spectrophotometry may be applied in performing direct studies on the intact microcapsules. The high activity yield of urease microcapsules relative to the mass of entrapped enzyme (92.5%) indicated minimal effects of mass transfer limitation. The mass of active urease incorporated into the nylon membrane represented 6% of the encapsulated enzyme activity. The soluble intracapsular enzyme fraction (94%) was released into solution upon rupture of the membrane. A complete mass and activity balance of the encapsulated enzyme was achieved.

  1. Preparation and evaluation of antifungal efficacy of griseofulvin loaded deformable membrane vesicles in optimized guinea pig model of Microsporum canis--dermatophytosis.

    Science.gov (United States)

    Aggarwal, Nidhi; Goindi, Shishu

    2012-11-01

    The present study is aimed at the encapsulation of griseofulvin in the deformable membrane vesicles (DMVs) for dermal delivery. Presently, griseofulvin is available only in conventional oral dosage forms that suffer from the issues of poor and highly variable bioavailability, numerous systemic side effects and long duration of treatment. Multi-lamellar drug-loaded DMVs of griseofulvin (Indian Patent Application 208/DEL/2009) were prepared by thin-film hydration method and were optimized for type and concentration of edge activator (EA). The optimized formulation was evaluated for vesicular shape, size, drug entrapment efficiency, drug content, pH, stability, spreadability, ex vivo skin permeation, dermatokinetics, skin sensitivity, in vitro antifungal assay and in vivo antifungal activity against Microsporum canis using guinea pig model for dermatophytosis. The optimized DMVs illustrated remarkably higher drug permeation and skin retention when compared with liposomes. A complete clinical and mycological cure was observed in animals treated with topical griseofulvin formulation in 10 days. The formulation was observed to be non-sensitizing, histopathologically safe, and stable at 5±3 °C, 25±2 °C and 40±2 °C for a period of six months. The results indicated that the topical formulation of DMVs of griseofulvin could be utilized as an alternative to reduce the encumbrance of conventional oral formulations. Copyright © 2012 Elsevier B.V. All rights reserved.

  2. Membrane dynamics

    DEFF Research Database (Denmark)

    Bendix, Pól Martin

    2015-01-01

    Current topics include membrane-protein interactions with regard to membrane deformation or curvature sensing by BAR domains. Also, we study the dynamics of membrane tubes of both cells and simple model membrane tubes. Finally, we study membrane phase behavior which has important implications...

  3. Modelling of proteins in membranes

    DEFF Research Database (Denmark)

    Sperotto, Maria Maddalena; May, S.; Baumgaertner, A.

    2006-01-01

    This review describes some recent theories and simulations of mesoscopic and microscopic models of lipid membranes with embedded or attached proteins. We summarize results supporting our understanding of phenomena for which the activities of proteins in membranes are expected to be significantly...... oppositely charged lipid membranes, lipid-induced tilting of proteins embedded in lipid bilayers, protein-induced bilayer deformations, protein insertion and assembly, and lipid-controlled functioning of membrane proteins....

  4. Lipid protrusions membrane softness, and enzymatic activity

    DEFF Research Database (Denmark)

    Jensen, Morten Østergaard; Høyrup, P.; Callisen, T.H.

    2004-01-01

    The activity of phospholipase A(2) on lipid bilayers displays a characteristic lag burst behavior that has previously been shown to reflect the physical properties of the substrate. It has remained unclear which underlying molecular mechanism is responsible for this phenomenon. We propose here...... protrusion modes and mechanical softness of phospholipid bilayers and on the other side the activity of enzymes acting on lipid bilayers composed of different unsaturated lipids. Specifically, our experiments show a correlation between the bilayer bending rigidity and the apparent Arrhenius activation energy...

  5. Active zone impact on deformation state of non-rigid pavement

    Directory of Open Access Journals (Sweden)

    Mandula Ján

    2014-06-01

    Full Text Available The paper deals with the design of non-rigid pavement, with emphasis on the effect of active zone on its deformation state. The concepts of determination of active zone are described. The results of numerical modelling of pavement laying on elastic subgrade are presented in the paper

  6. Activated sludge filterability and full-scale membrane bioreactor operation

    NARCIS (Netherlands)

    Krzeminski, P.

    2013-01-01

    Despite continuous developments in the field of MBR technology, membrane fouling together with the associated energy demand and related costs issues remain major challenges. The efficiency of the filtration process in an MBR is governed by the activated sludge filterability, which is still limitedly

  7. Flippase activity in proteoliposomes reconstituted with Spinacea oleracea endoplasmic reticulum membrane proteins: evidence of biogenic membrane flippase in plants.

    Science.gov (United States)

    Sahu, Santosh Kumar; Gummadi, Sathyanarayana N

    2008-09-30

    Phospholipid translocation (flip-flop) in biogenic (self-synthesizing) membranes such as the endoplasmic reticulum of eukaryotic cells (rat liver) and bacterial cytoplasmic membranes is a fundamental step in membrane biogenesis. It is known that flip-flop in these membranes occurs without a metabolic energy requirement, bidirectionally with no specificity for phospholipid headgroup. In this study, we demonstrate for the first time ATP-independent flippase activity in endoplasmic reticulum membranes of plants using spinach as a model system. For this, we generated proteoliposomes from a Triton X-100 extract of endoplasmic reticulum membranes of spinach and assayed them for flippase activity using fluorescently labeled phospholipids. The half-time for flipping was found to be 0.7-1.0 min. We also show that (a) proteoliposomes can flip fluorescently labeled analogues of phosphatidylcholine and phosphatidylethanolamine, (b) flipping activity is protein-mediated, (c) more than one class of lipid translocator (flippase) is present in spinach membranes, based on the sensitivity to protease and protein-modifying reagents, and (d) translocation of PC and PE is affected differently upon treatment with protease and protein-modifying reagents. Ca (2+)-dependent scrambling activity was not observed in the vesicles reconstituted from plant ER membranes, ruling out the possibility of the involvement of scramblase in translocation of phospholipids. These results suggest the existence of biogenic membrane flippases in plants and that the mechanism of membrane biogenesis is similar to that found in animals.

  8. Electrical resonance of Amphotericin B channel activity in lipidic membranes

    Science.gov (United States)

    Récamier, Karla S.; Ortega-Blake, Iván; Parmananda, P.

    2017-05-01

    In our previous work [J. Membrane Biol. 237, 31 (2010)], we showed the dependence of the time average conductance of Nystatin channels as a function of the applied potential. Specifically, it was observed that greater potential induced enhanced channel activity. This indicates that the supramolecular structure could be stabilized by a large field, possibly by giving a preferential orientation to the monomers. In the present work, we entertain the notion that the process of pore formation in the lipidic membranes has an underlying deterministic component. To verify this hypothesis, experiments were performed under potentio-dynamic conditions, i.e., a square train of pulses of different frequencies (0.05-2 Hz) were applied to a 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine membrane having 30 mol. % cholesterol and the presence of 35 μM Amphotericin B. An emergence of a resonant frequency, in the present experiments, is tantamount to observing fingerprints of determinism in the activity of these channels in lipidic membranes.

  9. Activity of platelet hemostasis in children with spinal deformities.

    Science.gov (United States)

    Medvedev, I N; Lapshina, E V; Zavalishina, S Yu

    2010-10-01

    An increase of adhesion and aggregation functions of platelets in vivo and in vitro was detected in 5-6-year-old children with scoliosis. These disorders were caused by hyperproduction of von Willebrand's factor in the vascular wall and intensification of thromboxane production in blood platelets. Activation of thromboxane formation is the main cause of platelet hyperactivity in children with scoliosis. Correction of platelet hemostasis may include pathogenetically substantiated complex of therapeutic exercises, swimming, and massage.

  10. Comparative performance between intermittently cyclic activated sludge-membrane bioreactor and anoxic/aerobic-membrane bioreactor.

    Science.gov (United States)

    Wang, Yu-Lan; Yu, Shui-Li; Shi, Wen-Xin; Bao, Rui-Ling; Zhao, Qing; Zuo, Xing-Tao

    2009-09-01

    A process of intermittently cyclic activated sludge-membrane bioreactor (ICAS-MBR) was developed to enhance the performance of biological phosphorus removal (EBPR), which was constructed under sequencing anoxic/anaerobic/aerobic condition. The performance between ICAS-MBR and conventional anoxic/aerobic-membrane bioreactor (A/O-MBR) in terms of phosphorus removal, nitrification performance and microbial aspects were investigated in this study. The experimental results indicated that the phosphorus removal efficiency in ICAS-MBR process increased from 65% to 83% when compared with A/O-MBR. It was also found that the COD removal efficiencies of the two processes were over 94%, and NH(4)(+)-N and TN average removal efficiencies were 96% and 69% in ICAS-MBR and 96% and 78% in A/O-MBR, respectively. Furthermore, micrographs observation obtained confirmed the succession and diversification of microorganisms in the two systems followed a similar regularity.

  11. Crustal Deformation around Zhangjiakou-Bohai Seismically Active Belt

    Science.gov (United States)

    Jin, H.; Fu, G.; Kato, T.

    2011-12-01

    Zhangjiakou-Bohai belt is a seismically active belt located in Northern China around Beijing, the capital of China. Near such a belt many great earthquakes occurred in the past centuries (e.g. the 1976 Tanshan Ms7.8 earthquake, the 1998 Zhangbei Ms6.2 earthquake, etc). Chinese Government established dense permanent and regional Global Positioning System (GPS) stations in and near the area. We collected and analyzed all the GPS observation data between 1999 and 2009 around Zhangjiakou-Bohai seismic belt, and obtained velocities at 143 stations. At the same time we investigated Zhangjiakou-Bohai belt slip rate for three profiles from northwest to southeast, and constructed a regional strain field on the Zhangjiakou-Bohai seismic belt region by least-square collocation. Based on the study we found that: 1) Nowadays the Zhangjiakou-Bohai seismic belt is creeping with left-lateral slip rate of 2.0mm~2.4mm/a, with coupling depth of 35~50km; 2) In total, the slip and coupling depth of the northwestern seismic belt is less than the one of southeast side; 3) The maximum shear strain is about 3×10-8 at Beijing-Tianjin-Tangshan area.

  12. Use of polyurethane foam deformation sensor to record respiratory activity

    Science.gov (United States)

    Bredov, V. I.; Baranov, V. S.

    1980-05-01

    The sensor developed has some substantial advantages over other known types. It is highly sensitive over a wide range of strain loads. The level of the output signal is linearly related to the force exerted on it, and it is sufficient for direct recording without using amplifiers of electric signals. The sensor is based on elastic, spongy material, polyurethane foam (porolon) with current-conducting material on the pore surface, current-conducting carbon black or electrode paste. The elastic properties of the sensor are built in the actual base of the strain-sensitive element, which simplifies the construction substantially and increases the reliability of the unit. In order to test the possibility of using this sensor to examine respiratory function, human pneumograms were recorded with the subject in a calm state along with the respiratory activity of experimental animals (dogs). Samples of the respiratory curve are shown. The simplicity of design of the sensor makes it possible to use it in various physiological experiments.

  13. Erythrocyte membrane stabilization effect and antioxidant activity of methyl methacrylate

    International Nuclear Information System (INIS)

    Popov, B.

    2004-01-01

    Methyl methacrylate (MMK) is a synthetic product with mild impact on human health that is not well studied on cellular basis. Here, human erythrocytes were used to investigate the effects MMK exerts on acid and heat-induced hemolysis. Biphasic effect of MMK was observed for acid-induced hemolysis; i.e., protection at low (0 - 0.05% v/v) and stimulation at higher (0.1- 0.4% v/v) concentrations. The maximal protective effect was produced at 0.03% (v/v). At this concentration MMK increased the temperatures of heat denaturation of erythrocyte membrane proteins, spectrin and integral proteins, by about 2 0 C and inhibited the heat-induced hemolysis by 20 %. This membrane stabilization effect of MMK is similar to that produced by some anti-inflammatory and antirheumatic drugs. The increased acid resistance possibly indicated anti-oxidant properties of MMK. The nonenzymatic antioxidant activity test evidenced that MMK has no superoxide dismutase-like activity but demonstrates strong catalase-like activity (about 900 kU/mmol at 0.05-0.1 mmol/l concentration). The results indicate that at low concentration MMK exerts benign effect on cellular membrane that could find therapeutic usage. (author)

  14. Active deformation processes across a megathrust-segment boundary, south-central Chile

    OpenAIRE

    M. Moreno; Juergen Klotz; D. Melnick; J. Bolte; Helmut Echtler; K. Bataille

    2009-01-01

    The south-central Chile margin is an active plate boundary where a variety of tectonic processes, including postseismic mantle relaxation, interseismic strain accumulation, sliver motions and crustal faulting are documented. GPS data and finite-element models with complex geometries are presented to gain insight into the active deformation in the vicinity of two megathrust-earthquake segments: the Valdivia and Concepción rupture zones. GPS vectors are heterogeneously distributed in two domain...

  15. DEFORMATION WAVES AS A TRIGGER MECHANISM OF SEISMIC ACTIVITY IN SEISMIC ZONES OF THE CONTINENTAL LITHOSPHERE

    Directory of Open Access Journals (Sweden)

    S. I. Sherman

    2013-01-01

    Full Text Available Deformation waves as a trigger mechanism of seismic activity and migration of earthquake foci have been under discussion by researchers in seismology and geodynamics for over 50 years. Four sections of this article present available principal data on impacts of wave processes on seismicity and new data. The first section reviews analytical and experimental studies aimed at identification of relationships between wave processes in the lithosphere and seismic activity manifested as space-and-time migration of individual earthquake foci or clusters of earthquakes. It is concluded that with a systematic approach, instead of using a variety of terms to denote waves that trigger seismic process in the lithosphere, it is reasonable to apply the concise definition of ‘deformation waves’, which is most often used in fact.The second section contains a description of deformation waves considered as the trigger mechanism of seismic activity. It is concluded that a variety of methods are applied to identify deformation waves, and such methods are based on various research methods and concepts that naturally differ in sensitivity concerning detection of waves and/or impact of the waves on seismic process. Epicenters of strong earthquakes are grouped into specific linear or arc-shaped systems, which common criterion is the same time interval of the occurrence of events under analysis. On site the systems compose zones with similar time sequences, which correspond to the physical notion of moving waves (Fig. 9. Periods of manifestation of such waves are estimated as millions of years, and a direct consideration of the presence of waves and wave parameters is highly challenging. In the current state-of-the-art, geodynamics and seismology cannot provide any other solution yet.The third section presents a solution considering record of deformation waves in the lithosphere. With account of the fact that all the earthquakes with М≥3.0 are associated with

  16. Two-dimensional strain-hardening membrane model for large deformation behavior of multiple red blood cells in high shear conditions.

    Science.gov (United States)

    Ye, Swe Soe; Ng, Yan Cheng; Tan, Justin; Leo, Hwa Liang; Kim, Sangho

    2014-05-13

    Computational modeling of Red Blood Cell (RBC) flow contributes to the fundamental understanding of microhemodynamics and microcirculation. In order to construct theoretical RBC models, experimental studies on single RBC mechanics have presented a material description for RBC membranes based on their membrane shear, bending and area moduli. These properties have been directly employed in 3D continuum models of RBCs but practical flow analysis with 3D models have been limited by their computationally expensive nature. As such, various researchers have employed 2D models to efficiently and qualitatively study microvessel flows. Currently, the representation of RBC dynamics using 2D models is a limited methodology that breaks down at high shear rates due to excessive and unrealistic stretching. We propose a localized scaling of the 2D elastic moduli such that it increases with RBC local membrane strain, thereby accounting for effects such as the Poisson effect and membrane local area incompressibility lost in the 2D simplification. Validation of our 2D Large Deformation (2D-LD) RBC model was achieved by comparing the predicted RBC deformation against the 3D model from literature for the case of a single RBC in simple shear flow under various shear rates (dimensionless shear rate G = 0.05, 0.1, 0.2, 0.5). The multi-cell flow of RBCs (38% Hematocrit) in a 20 μm width microchannel under varying shear rates (50, 150, 150 s-1) was then simulated with our proposed model and the popularly-employed 2D neo-Hookean model in order to evaluate the efficacy of our proposed 2D-LD model. The validation set indicated similar RBC deformation for both the 2D-LD and the 3D models across the studied shear rates, highlighting the robustness of our model. The multi-cell simulation indicated that the 2D neo-Hookean model predicts noodle-like RBC shapes at high shear rates (G = 0.5) whereas our 2D-LD model maintains sensible RBC deformations. The ability of the 2D-LD model to

  17. Photosynthesis Activates Plasma Membrane H+-ATPase via Sugar Accumulation.

    Science.gov (United States)

    Okumura, Masaki; Inoue, Shin-Ichiro; Kuwata, Keiko; Kinoshita, Toshinori

    2016-05-01

    Plant plasma membrane H(+)-ATPase acts as a primary transporter via proton pumping and regulates diverse physiological responses by controlling secondary solute transport, pH homeostasis, and membrane potential. Phosphorylation of the penultimate threonine and the subsequent binding of 14-3-3 proteins in the carboxyl terminus of the enzyme are required for H(+)-ATPase activation. We showed previously that photosynthesis induces phosphorylation of the penultimate threonine in the nonvascular bryophyte Marchantia polymorpha However, (1) whether this response is conserved in vascular plants and (2) the process by which photosynthesis regulates H(+)-ATPase phosphorylation at the plasma membrane remain unresolved issues. Here, we report that photosynthesis induced the phosphorylation and activation of H(+)-ATPase in Arabidopsis (Arabidopsis thaliana) leaves via sugar accumulation. Light reversibly phosphorylated leaf H(+)-ATPase, and this process was inhibited by pharmacological and genetic suppression of photosynthesis. Immunohistochemical and biochemical analyses indicated that light-induced phosphorylation of H(+)-ATPase occurred autonomously in mesophyll cells. We also show that the phosphorylation status of H(+)-ATPase and photosynthetic sugar accumulation in leaves were positively correlated and that sugar treatment promoted phosphorylation. Furthermore, light-induced phosphorylation of H(+)-ATPase was strongly suppressed in a double mutant defective in ADP-glucose pyrophosphorylase and triose phosphate/phosphate translocator (adg1-1 tpt-2); these mutations strongly inhibited endogenous sugar accumulation. Overall, we show that photosynthesis activated H(+)-ATPase via sugar production in the mesophyll cells of vascular plants. Our work provides new insight into signaling from chloroplasts to the plasma membrane ion transport mechanism. © 2016 American Society of Plant Biologists. All Rights Reserved.

  18. Influence of van der Waals contact forces on the deformation mechanics of thin flexible membranes assembled from metallic or semiconducting single-wall carbon nanotubes

    Science.gov (United States)

    Hobbie, Erik K.; Harris, John; Iyer, Swathi; Huh, Ji Yeon; Fagan, Jeffrey A.; Hudson, Steven D.; Stafford, Christopher M.

    2011-03-01

    Thin membranes of single-wall carbon nanotubes (SWCNTs) assembled from either metallic or semiconducting SWCNTs are subjected to the compressive strains imposed by a stretched elastic substrate, and the mechanical characteristics of the membranes are inferred from the topography of the wrinkling instability that emerges. By depositing comparable films on quartz, we also use optical (UV-Vis-NIR) absorption spectroscopy to compute the effective London dispersion spectra of the purified materials, and from these we compute the attractive part of the van der Waals potential between nanotubes of identical electronic type as a function of separation and relative orientation. We find significant differences in the strength and shape of the contact potential depending on electronic type, which in turn are evident in the modulus and yield strain measured from the deformation of the films. Supported by the NSF through CMMI-0969155 and the DOE through DE-FG36-08GO88160.

  19. Calcium Modulation of Plant Plasma Membrane-Bound Atpase Activities

    Science.gov (United States)

    Caldwell, C.

    1983-01-01

    The kinetic properties of barley enzyme are discussed and compared with those of other plants. Possibilities for calcium transport in the plasma membrane by proton pump and ATPase-dependent calcium pumps are explored. Topics covered include the ph phase of the enzyme; high affinity of barley for calcium; temperature dependence, activation enthalpy, and the types of ATPase catalytic sites. Attention is given to lipids which are both screened and bound by calcium. Studies show that barley has a calmodulin activated ATPase that is found in the presence of magnesium and calcium.

  20. Thin Film Polyamide Membranes with Photoresponsive Antibacterial Activity

    KAUST Repository

    Duong, Phuoc H. H.

    2017-08-09

    Membranes containing a photosensitizer molecule as part of the selective layer are proposed with demonstrated anti-biofouling activity. For the membrane preparation, mixtures of an amine-functionalized photosensitizer molecule, (5,10,15,20-(tetra-4-aminophenyl)porphyrin) and m-phenylene diamine (MPD) reacted with trimesoyl chloride (TMC) by interfacial polymerization to form thin polyamide films on top of an asymmetric porous support. A highly permeable membrane (35.4 Lm−2h−1bar−1) with 99 % rejection of Brilliant Blue R (826 g/mol) was obtained using 0.25 wt% porphyrin and 0.75 wt% MPD as amine monomers. Under visible light exposure, singlet oxygen (1O2) is generated in the porphyrin containing-polyamide film, reaching the bacteria in the feed by diffusion and enhancing the biofouling resistance and anti-microbial activity. Anti-biofouling and anti-microbial photoactivity in solution are demonstrated on Staphylococcus aureus at different porphyrin concentrations and light exposure time.

  1. QCM-D fingerprinting of membrane-active peptides.

    Science.gov (United States)

    McCubbin, George A; Praporski, Slavica; Piantavigna, Stefania; Knappe, Daniel; Hoffmann, Ralf; Bowie, John H; Separovic, Frances; Martin, Lisandra L

    2011-04-01

    The increasing prevalence of antibiotic-resistant bacteria is becoming a public health crisis. Antimicrobial peptides (AMPs) are a promising solution, because bacterial resistance is less likely. Quartz crystal microbalance with dissipation monitoring (QCM-D) is a versatile and valuable technique for investigation of these peptides. This article looks at the different approaches to the interpretation of QCM-D data, showing how to extract the maximum information from the data. Five AMPs of diverse charge, length and activity are used as case studies: caerin 1.1 wild-type, two caerin 1.1 mutants (Gly15Gly19-caerin 1.1 and Ala15Ala19-caerin 1.1), aurein 1.2 and oncocin. The interaction between the AMP and a 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) membrane is analysed inter alia using frequency-dissipation plots (∆f-∆D plots) to ascertain the mechanism of action of the AMP. The ∆f-∆D plot can then be used to provide a fingerprint for the AMP-membrane interaction. Building up a database of these fingerprints for all known AMPs will enable the relationship between AMP structure and membrane activity to be better understood, hopefully leading to the future development of antibiotics without bacterial resistance.

  2. How To Tackle the Issues in Free Energy Simulations of Long Amphiphiles Interacting with Lipid Membranes: Convergence and Local Membrane Deformations

    DEFF Research Database (Denmark)

    Filipe, H. A. L.; Moreno, M. J.; Rog, T.

    2014-01-01

    One of the great challenges in membrane biophysics is to find a means to foster the transport of drugs across complex membrane structures. In this spirit, we elucidate methodological challenges associated with free energy computations of complex chainlike molecules across lipid membranes....... As an appropriate standard molecule to this end, we consider 7-nitrobenz-2-oxa-1,3-diazol-4-yl-labeled fatty amine, NBD-C-n, which is here dealt with as a homologous series with varying chain lengths. We found the membrane-water interface region to be highly sensitive to details in free energy computations. Despite...... of radius 1.7 nm from the amphiphile. Importantly, the free energy results given by PGC were found to be qualitatively consistent with experimental data, while the PGD results were not. We conclude that with long amphiphiles there is reason for concern with regard to computations of their free energy...

  3. Antiviral activity of squalamine: Role of electrostatic membrane binding

    Science.gov (United States)

    Beckerman, Bernard; Qu, Wei; Mishra, Abhijit; Zasloff, Michael; Wong, Gerard; Luijten, Erik

    2012-02-01

    Recent workootnotetextM. Zasloff et al., Proc. Nat. Acad. Sci. (USA) 108, 15978 (2011). has demonstrated that squalamine, a molecule found in the liver of sharks, exhibits broad-spectrum antiviral properties. It has been proposed that this activity results from the charge-density matching of squalamine and phospholipid membranes, causing squalamine to bind to membranes and displace proteins such as Rac1 that are crucial for the viral replication cycle. Here we investigate this hypothesis by numerical simulation of a coarse-grained model for the competition between Rac1 and squalamine in binding affinity to a flat lipid bilayer. We perform free-energy calculations to test the ability of squalamine to condense stacked bilayer systems and thereby displace bulkier Rac1 molecules. We directly compare our findings to small-angle x-ray scattering results for the same setup.

  4. A parallel interaction potential approach coupled with the immersed boundary method for fully resolved simulations of deformable interfaces and membranes

    Science.gov (United States)

    Spandan, Vamsi; Meschini, Valentina; Ostilla-Mónico, Rodolfo; Lohse, Detlef; Querzoli, Giorgio; de Tullio, Marco D.; Verzicco, Roberto

    2017-11-01

    In this paper we show and discuss how the deformation dynamics of closed liquid-liquid interfaces (for example drops and bubbles) can be replicated with use of a phenomenological interaction potential model. This new approach to simulate liquid-liquid interfaces is based on the fundamental principle of minimum potential energy where the total potential energy depends on the extent of deformation of a spring network distributed on the surface of the immersed drop or bubble. Simulating liquid-liquid interfaces using this model require computing ad-hoc elastic constants which is done through a reverse-engineered approach. The results from our simulations agree very well with previous studies on the deformation of drops in standard flow configurations such as a deforming drop in a shear flow or cross flow. The interaction potential model is highly versatile, computationally efficient and can be easily incorporated into generic single phase fluid solvers to also simulate complex fluid-structure interaction problems. This is shown by simulating flow in the left ventricle of the heart with mechanical and natural mitral valves where the imposed flow, motion of ventricle and valves dynamically govern the behaviour of each other. Results from these simulations are compared with ad-hoc in-house experimental measurements. Finally, we present a simple and easy to implement parallelisation scheme, as high performance computing is unavoidable when studying large scale problems involving several thousands of simultaneously deforming bodies in highly turbulent flows.

  5. Crustal deformation and volcanic earthquakes associated with the recent volcanic activity of Iwojima Volcano, Japan

    Science.gov (United States)

    Ueda, H.; Fujita, E.; Tanada, T.

    2013-12-01

    Iwojima is an active volcanic island located within a 10 km wide submarine caldera about 1250 km to the south of Tokyo, Japan. The seismometer and GPS network of National Research Institute for Earth Science and Disaster Prevention (NIED) in Iwojima has observed a repeating island wide uplift more than 1 m associated with large number of volcanic earthquakes every several years. During 2006-2012, we observed more than 20000 volcanic earthquakes and an uplift of about 3 m, and precursory volcanic earthquakes and rapid crustal deformation just before the small submarine eruption near the northern coast of Iwojima in April 2012. In a restless volcano such as Iwojima, it is important issue to distinguish whether rapid crustal deformation and intense earthquake activity lead to an eruption or not. According to a long period geodetic observation by Ukawa et al. (2006), the crustal deformation of Iwojima can be classify into 2 phases. The first is an island wide large uplift centering on Motoyama area (the eastern part of the island, the center of the caldera), and the second is contraction and subsidence at local area centering on Motoyama and uplift around that area. They are interpreted by superposition of crustal deformations by a shallow contraction source and a deep seated inflation source beneath Motoyama. The earthquake activity of Iwojima highly correlates with the island wide large uplift, suggesting the earthquakes are almost controlled by a magma accumulation into a deep seated magma chamber. In contrast to the activity, the precursory activity of the eruption in 2012 is deviated from the correlation. The rapid crustal deformation just before and after the eruption in 2012 can be interpreted by rapid inflation and deflation of a shallow sill source about 1km deep, respectively, suggesting that it was caused by a shallow hydrothermal activity. The result shows that we can probably distinguish an abnormal activity related with a volcanic eruption when we observe

  6. Dimerization Efficiency of Canine Distemper Virus Matrix Protein Regulates Membrane-Budding Activity.

    Science.gov (United States)

    Bringolf, Fanny; Herren, Michael; Wyss, Marianne; Vidondo, Beatriz; Langedijk, Johannes P; Zurbriggen, Andreas; Plattet, Philippe

    2017-08-15

    Paramyxoviruses rely on the matrix (M) protein to orchestrate viral assembly and budding at the plasma membrane. Although the mechanistic details remain largely unknown, structural data suggested that M dimers and/or higher-order oligomers may facilitate membrane budding. To gain functional insights, we employed a structure-guided mutagenesis approach to investigate the role of canine distemper virus (CDV) M protein self-assembly in membrane-budding activity. Three six-alanine-block (6A-block) mutants with mutations located at strategic oligomeric positions were initially designed. While the first one includes residues potentially residing at the protomer-protomer interface, the other two display amino acids located within two distal surface-exposed α-helices proposed to be involved in dimer-dimer contacts. We further focused on the core of the dimeric interface by mutating asparagine 138 (N138) to several nonconservative amino acids. Cellular localization combined with dimerization and coimmunopurification assays, performed under various denaturing conditions, revealed that all 6A-block mutants were impaired in self-assembly and cell periphery accumulation. These phenotypes correlated with deficiencies in relocating CDV nucleocapsid proteins to the cell periphery and in virus-like particle (VLP) production. Conversely, all M-N138 mutants remained capable of self-assembly, though to various extents, which correlated with proper accumulation and redistribution of nucleocapsid proteins at the plasma membrane. However, membrane deformation and VLP assays indicated that the M-N138 variants exhibiting the most reduced dimerization propensity were also defective in triggering membrane remodeling and budding, despite proper plasma membrane accumulation. Overall, our data provide mechanistic evidence that the efficiency of CDV M dimerization/oligomerization governs both cell periphery localization and membrane-budding activity. IMPORTANCE Despite the availability of

  7. Modulation of Erythrocyte Plasma Membrane Redox System Activity by Curcumin

    Directory of Open Access Journals (Sweden)

    Prabhakar Singh

    2016-01-01

    Full Text Available Plasma membrane redox system (PMRS is an electron transport chain system ubiquitously present throughout all cell types. It transfers electron from intracellular substrates to extracellular acceptors for regulation of redox status. Curcumin, isolated from Curcuma longa, has modulatory effects on cellular physiology due to its membrane interaction ability and antioxidant potential. The present study investigates the effect of curcumin on PMRS activity of erythrocytes isolated from Wistar rats in vitro and in vivo and validated through an in silico docking simulation study using Molegro Virtual Docker (MVD. Effects of curcumin were also evaluated on level of glutathione (GSH and the oxidant potential of plasma measured in terms of plasma ferric equivalent oxidative potentials (PFEOP. Results show that curcumin significantly (p<0.01 downregulated the PMRS activity in a dose-dependent manner. Molecular docking results suggest that curcumin interacts with amino acids at the active site cavity of cytochrome b5 reductase, a key constituent of PMRS. Curcumin also increased the GSH level in erythrocytes and plasma while simultaneously decreasing the oxidant potential (PFEOP of plasma. Altered PMRS activity and redox status are associated with the pathophysiology of several health complications including aging and diabetes; hence, the above finding may explain part of the role of curcumin in health beneficial effects.

  8. Cell volume and membrane stretch independently control K+ channel activity

    DEFF Research Database (Denmark)

    Bomholtz, Sofia Hammami; Willumsen, Niels J; Olsen, Hervør L

    2009-01-01

    A number of potassium channels including members of the KCNQ family and the Ca(2+) activated IK and SK, but not BK, are strongly and reversibly regulated by small changes in cell volume. It has been argued that this general regulation is mediated through sensitivity to changes in membrane stretch....... To test this hypothesis we have studied the regulation of KCNQ1 and BK channels after expression in Xenopus oocytes. Results from cell-attached patch clamp studies (approximately 50 microm(2) macropatches) in oocytes expressing BK channels demonstrate that the macroscopic volume-insensitive BK current...... that stretch and volume sensitivity can be considered two independent regulatory mechanisms....

  9. A parallel interaction potential approach coupled with the immersed boundary method for fully resolved simulations of deformable interfaces and membranes

    NARCIS (Netherlands)

    Arza, Vamsi Spandan; Meschini, Valentina; Ostilla-Mónico, Rodolfo; Lohse, Detlef; Querzoli, Giorgio; de Tullio, Marco D.; Verzicco, Roberto

    2017-01-01

    In this paper we show and discuss how the deformation dynamics of closed liquid–liquid interfaces (for example drops and bubbles) can be replicated with use of a phenomenological interaction potential model. This new approach to simulate liquid–liquid interfaces is based on the fundamental principle

  10. Active deformation processes across a megathrust-segment boundary, south-central Chile

    Science.gov (United States)

    Moreno, M. S.; Klotz, J.; Melnick, D.; Bolte, J.; Echtler, H.; Bataille, K.

    2009-04-01

    The south-central Chile margin is an active plate boundary where a variety of tectonic processes, including postseismic mantle relaxation, interseismic strain accumulation, sliver motions and crustal faulting are documented. GPS data and finite-element models with complex geometries are presented to gain insight into the active deformation in the vicinity of two megathrust-earthquake segments: the Valdivia and Concepción rupture zones. GPS vectors are heterogeneously distributed in two domains that follow the megathrust segments. Models which simulate only interseismic locking on the plate interface and postseismic relaxation after an uniform coseismic slip during the 1960 Valdivia earthquake (Mw=9.5) not fully reproduce the observed surface deformation. In order to distinguish between the main processes producing the regional-scale heterogeneity of surface velocities, we model: (1) the postseismic viscoelastic deformation induced by a non-uniform coseismic slip distribution, (2) the interseismic kinematic coupling, and (3) the interseismic effect of a crustal fault rooted in the plate interface. We find that interseismic locked asperities are spatially coincident with the historic earthquake rupture zones, which are separated by a sharp boundary of low (~ 50%) kinematic coupling. A regional pattern of clockwise rotations arise from postseismic mantle rebound at the overlap area of the rupture zones and extend over a broad segment between the forearc and the back-arc. Locally, counterclockwise block rotation is observed in a limited area south of the Lanalhue fault, and may be related to transpression at the northern leading edge of a forearc sliver. In the Concepción domain, models that include a dextral-reverse crustal fault better reproduce the GPS observations. Our study suggests that upper-plate crustal faults in addition to earthquake-cycle transients exert an important control on deformation processes at megathrust-segment boundaries.

  11. Mechanism of Action of a Membrane-Active Quinoline-Based Antimicrobial on Natural and Model Bacterial Membranes.

    Science.gov (United States)

    Hubbard, Alasdair T M; Barker, Robert; Rehal, Reg; Vandera, Kalliopi-Kelli A; Harvey, Richard D; Coates, Anthony R M

    2017-02-28

    HT61 is a quinoline-derived antimicrobial, which exhibits bactericidal potency against both multiplying and quiescent methicillin resistant and sensitive Staphylococcus aureus, and has been proposed as an adjunct for other antimicrobials to extend their usefulness in the face of increasing antimicrobial resistance. In this study, we have examined HT61's effect on the permeability of S. aureus membranes and whether this putative activity can be attributed to an interaction with lipid bilayers. Using membrane potential and ATP release assays, we have shown that HT61 disrupts the membrane enough to result in depolarization of the membrane and release of intercellular constituents at concentrations above and below the minimum inhibitory concentration of the drug. Utilizing both monolayer subphase injection and neutron reflectometry, we have shown that increasing the anionic lipid content of the membrane leads to a more marked effect of the drug. In bilayers containing 25 mol % phosphatidylglycerol, neutron reflectometry data suggest that exposure to HT61 increases the level of solvent in the hydrophobic region of the membrane, which is indicative of gross structural damage. Increasing the proportion of PG elicits a concomitant level of membrane damage, resulting in almost total destruction when 75 mol % phosphatidylglycerol is present. We therefore propose that HT61's primary action is directed toward the cytoplasmic membrane of Gram-positive bacteria.

  12. Active deformation in the northern Sierra de Valle Fértil, Sierras Pampeanas, Argentina

    Science.gov (United States)

    Ortiz, Gustavo; Alvarado, Patricia; Fosdick, Julie C.; Perucca, Laura; Saez, Mauro; Venerdini, Agostina

    2015-12-01

    The Western Sierras Pampeanas region in the San Juan Province is characterized by thick-skinned deformation with approximately N-S trending ranges of average heights of 2500 m and a high frequency occurrence of seismic activity. Its location to the east of the mainly thin-skinned tectonics of the Argentine Precordillera fold-and-thrust belt suggests that at 30°S, deformation is concentrated in a narrow zone involving these two morphostructural units. In this paper, we present new apatite (U-Th)/He results (AHe) across the northern part of the Sierra de Valle Fértil (around 30°S) and analyze them in a framework of thermochronologic available datasets. We found Pliocene AHe results for Carboniferous and Triassic strata in the northern Sierra de Valle Fértil consistent with the hypothesis of recent cooling and inferred erosional denudation concentrated along the northern end of this mountain range. Our analysis shows that this northern region may have evolved under different conditions than the central part of the Sierra de Valle Fértil. Previous studies have observed AHe ages consistent with Permian through Cretaceous cooling, indicating the middle part of the Sierra de Valle Fértil remained near surface before the Pampean slab subduction flattening process. Those studies also obtained ˜5 My cooling ages in the southern part of the Sierra de Valle Fértil, which are similar to our results in the northern end of the range. Taken together, these results suggest a pattern of young deformation in the northern and southern low elevation ends of the Sierra de Valle Fértil consistent with regions of high seismic activity, and Quaternary active faulting along the western-bounding thrust fault of the Sierra de Valle Fértil.

  13. Low resistivity and permeability in actively deforming shear zones on the San Andreas Fault at SAFOD

    Science.gov (United States)

    Morrow, Carolyn A.; Lockner, David A.; Hickman, Stephen H.

    2015-01-01

    The San Andreas Fault Observatory at Depth (SAFOD) scientific drillhole near Parkfield, California crosses the San Andreas Fault at a depth of 2.7 km. Downhole measurements and analysis of core retrieved from Phase 3 drilling reveal two narrow, actively deforming zones of smectite-clay gouge within a roughly 200 m-wide fault damage zone of sandstones, siltstones and mudstones. Here we report electrical resistivity and permeability measurements on core samples from all of these structural units at effective confining pressures up to 120 MPa. Electrical resistivity (~10 ohm-m) and permeability (10-21 to 10-22 m2) in the actively deforming zones were one to two orders of magnitude lower than the surrounding damage zone material, consistent with broader-scale observations from the downhole resistivity and seismic velocity logs. The higher porosity of the clay gouge, 2 to 8 times greater than that in the damage zone rocks, along with surface conduction were the principal factors contributing to the observed low resistivities. The high percentage of fine-grained clay in the deforming zones also greatly reduced permeability to values low enough to create a barrier to fluid flow across the fault. Together, resistivity and permeability data can be used to assess the hydrogeologic characteristics of the fault, key to understanding fault structure and strength. The low resistivities and strength measurements of the SAFOD core are consistent with observations of low resistivity clays that are often found in the principal slip zones of other active faults making resistivity logs a valuable tool for identifying these zones.

  14. Blood-group-Ii-active gangliosides of human erythrocyte membranes

    International Nuclear Information System (INIS)

    Feizi, T.; Childs, R.A.; Hakomori, S.-I.; Powell, M.E.

    1978-01-01

    More than ten new types of gangliosides, in addition to haematoside and sialosylparagloboside, were isolated from human erythrocyte membranes. These were separated by successive chromatographies on DAEA-Sephadex, on porous silica-gel columns and on thin-layer silica gel as acetylated compounds. Highly potent blood-group-Ii and moderate blood-group-H activities were demonstrated in some of the ganglioside fractions. The gangliosides incorporated into chlolesterol/phosphatidylcholine liposomes stoicheiometrically inhibited binding of anti-(blood-group-I and i) antibodies to a radioiodinated blood-group-Ii-active glycoprotein. The fraction with the highest blood-group-I activity, I(g) fraction, behaved like sialosyl-deca- to dodeca-glycosylceramides on t.l.c. Certain blood-group-I and most of the i-determinants were in partially or completely cryptic form and could be unmasked by sialidase treatment. Thus the I and i antigens, which are known to occur on internal structures of blood-group-ABH-active glycoproteins in secretions, also occur in the interior of the carbohydrate chains of erythrocyte gangliosides. (author)

  15. Red wine activates plasma membrane redox system in human erythrocytes.

    Science.gov (United States)

    Tedesco, Idolo; Moccia, Stefania; Volpe, Silvestro; Alfieri, Giovanna; Strollo, Daniela; Bilotto, Stefania; Spagnuolo, Carmela; Di Renzo, Massimo; Aquino, Rita P; Russo, Gian Luigi

    2016-01-01

    In the present study, we report that polyphenols present in red wine obtained by a controlled microvinification process are able to protect human erythrocytes from oxidative stress and to activate Plasma Membrane Redox System (PMRS). Human plasma obtained from healthy subjects was incubated in the presence of whole red wine at a concentration corresponding to 9.13-73 μg/ml gallic acid equivalents to verify the capacity to protect against hypochlorous acid (HOCl)-induced plasma oxidation and to minimize chloramine formation. Red wine reduced hemolysis and chloramine formation induced by HOCl of 40 and 35%, respectively. PMRS present on human erythrocytes transfers electrons from intracellular molecules to extracellular electron acceptors. We demonstrated that whole red wine activated PMRS activity in human erythrocytes isolated from donors in a dose-dependent manner with a maximum at about 70-100 μg/ml gallic acid equivalents. We also showed that red wine increased glutathione (GSH) levels and erythrocytic antioxidant capacity, measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) quenching assay. Furthermore, we reported that GSH played a crucial role in regulating PMRS activity in erythrocytes. In fact, the effect of iodoacetamide, an alkylating agent that induces depletion of intracellular GSH, was completely counteracted by red wine. Bioactive compounds present in red wine, such as gallic acid, resveratrol, catechin, and quercetin were unable to activate PMRS when tested at the concentrations normally present in aged red wines. On the contrary, the increase of PMRS activity was associated with the anthocyanin fraction, suggesting the capacity of this class of compounds to positively modulate PMRS enzymatic activity.

  16. Effect of activated carbon in PSF-PEI-Ag symmetric membrane

    OpenAIRE

    Said Khairul Anwar Mohamad; George Genevieve Gadung; Alipah Nurul Ain Mohamed; Ismail Nor Zakirah; Jama’in Ramizah Liyana

    2017-01-01

    Polysulfone (Psf) composite membrane consist of activated carbon, polyethyleneimine and silver nitrate was prepared by phase inversion. The activated carbon (AC) act as adsorbent to adsorb heavy metal present in synthetic waste water while polysulfone membrane act as support. Phase inversion was carried out on different composition of activated carbon from 0 to 0.9% while other component are remain constant. The surface morphology of composite membrane was characterized by scanning electron m...

  17. Contemporaneous ring fault activity and surface deformation at subsiding calderas studied using analogue experiments

    Science.gov (United States)

    Liu, Yuan-Kai; Ruch, Joël; Vasyura-Bathke, Hannes; Jónsson, Sigurjón

    2017-04-01

    Ground deformation analyses of several subsiding calderas have shown complex and overlapping deformation signals, with a broad deflation signal that affects the entire volcanic edifice and localized subsidence focused within the caldera. However, the relation between deep processes at subsiding calderas, including magmatic sources and faulting, and the observed surface deformation is still debated. Several recent examples of subsiding calderas in the Galápagos archipelago and at the Axial seamount in the Pacific Ocean indicate that ring fault activity plays an important role not only during caldera collapse, but also during initial stages of caldera subsidence. Nevertheless, ring fault activity has rarely been integrated into numerical models of subsiding calderas. Here we report on sandbox analogue experiments that we use to study the processes involved from an initial subsidence to a later collapse of calderas. The apparatus is composed of a subsiding half piston section connected to the bottom of a glass box and driven by a motor to control its subsidence. We analyze at the same time during the subsidence the 3D displacement at the model surface with a laser scanner and the 2D ring fault evolution on the side of the model (cross-section) with a side-view digital camera. We further use PIVLab, a time-resolved digital image correlation software tool, to extract strain and velocity fields at both the surface and in cross-section. This setup allows to track processes acting at depth and assess their relative importance as the collapse evolves. We further compare our results with the examples observed in nature as well as with numerical models that integrate ring faults.

  18. Basement Structure and Styles of Active Tectonic Deformation in Central Interior Alaska

    Science.gov (United States)

    Dixit, N.; Hanks, C.

    2017-12-01

    Central Interior Alaska is one of the most seismically active regions in North America, exhibiting a high concentration of intraplate earthquakes approximately 700 km away from the southern Alaska subduction zone. Based on increasing seismological evidence, intraplate seismicity in the region does not appear to be uniformly distributed, but concentrated in several discrete seismic zones, including the Nenana basin and the adjacent Tanana basin. Recent seismological and neotectonics data further suggests that these seismic zones operate within a field of predominantly pure shear driven primarily by north-south crustal shortening. Although the location and magnitude of the seismic activity in both basins are well defined by a network of seismic stations in the region, the tectonic controls on intraplate earthquakes and the heterogeneous nature of Alaska's continental interior remain poorly understood. We investigated the current crustal architecture and styles of tectonic deformation of the Nenana and Tanana basins using existing geological, geophysical and geochronological datasets. The results of our study demonstrate that the basements of the basins show strong crustal heterogeneity. The Tanana basin is a relatively shallow (up to 2 km) asymmetrical foreland basin with its southern, deeper side controlled by the northern foothills of the central Alaska Range. Northeast-trending strike-slip faults within the Tanana basin are interpreted as a zone of clockwise crustal block rotation. The Nenana basin has a fundamentally different geometry; it is a deep (up to 8 km), narrow transtensional pull-apart basin that is deforming along the left-lateral Minto Fault. This study identifies two distinct modes of tectonic deformation in central Interior Alaska at present, and provides a basis for modeling the interplay between intraplate stress fields and major structural features that potentially influence the generation of intraplate earthquakes in the region.

  19. The Recombinant Sea Urchin Immune Effector Protein, rSpTransformer-E1, Binds to Phosphatidic Acid and Deforms Membranes

    Directory of Open Access Journals (Sweden)

    Cheng Man Lun

    2017-05-01

    Full Text Available The purple sea urchin, Strongylocentrotus purpuratus, possesses a sophisticated innate immune system that functions without adaptive capabilities and responds to pathogens effectively by expressing the highly diverse SpTransformer gene family (formerly the Sp185/333 gene family. The swift gene expression response and the sequence diversity of SpTransformer cDNAs suggest that the encoded proteins have immune functions. Individual sea urchins can express up to 260 distinct SpTransformer proteins, and their diversity suggests that different versions may have different functions. Although the deduced proteins are diverse, they share an overall structure of a hydrophobic leader, a glycine-rich N-terminal region, a histidine-rich region, and a C-terminal region. Circular dichroism analysis of a recombinant SpTransformer protein, rSpTransformer-E1 (rSpTrf-E1 demonstrates that it is intrinsically disordered and transforms to α helical in the presence of buffer additives and binding targets. Although native SpTrf proteins are associated with the membranes of perinuclear vesicles in the phagocyte class of coelomocytes and are present on the surface of small phagocytes, they have no predicted transmembrane region or conserved site for glycophosphatidylinositol linkage. To determine whether native SpTrf proteins associate with phagocyte membranes through interactions with lipids, when rSpTrf-E1 is incubated with lipid-embedded nylon strips, it binds to phosphatidic acid (PA through both the glycine-rich region and the histidine-rich region. Synthetic liposomes composed of PA and phosphatidylcholine show binding between rSpTrf-E1 and PA by fluorescence resonance energy transfer, which is associated with leakage of luminal contents suggesting changes in lipid organization and perhaps liposome lysis. Interactions with liposomes also change membrane curvature leading to liposome budding, fusion, and invagination, which is associated with PA clustering induced

  20. The Recombinant Sea Urchin Immune Effector Protein, rSpTransformer-E1, Binds to Phosphatidic Acid and Deforms Membranes.

    Science.gov (United States)

    Lun, Cheng Man; Samuel, Robin L; Gillmor, Susan D; Boyd, Anthony; Smith, L Courtney

    2017-01-01

    The purple sea urchin, Strongylocentrotus purpuratus , possesses a sophisticated innate immune system that functions without adaptive capabilities and responds to pathogens effectively by expressing the highly diverse SpTransformer gene family (formerly the Sp185/333 gene family). The swift gene expression response and the sequence diversity of SpTransformer cDNAs suggest that the encoded proteins have immune functions. Individual sea urchins can express up to 260 distinct SpTransformer proteins, and their diversity suggests that different versions may have different functions. Although the deduced proteins are diverse, they share an overall structure of a hydrophobic leader, a glycine-rich N-terminal region, a histidine-rich region, and a C-terminal region. Circular dichroism analysis of a recombinant SpTransformer protein, rSpTransformer-E1 (rSpTrf-E1) demonstrates that it is intrinsically disordered and transforms to α helical in the presence of buffer additives and binding targets. Although native SpTrf proteins are associated with the membranes of perinuclear vesicles in the phagocyte class of coelomocytes and are present on the surface of small phagocytes, they have no predicted transmembrane region or conserved site for glycophosphatidylinositol linkage. To determine whether native SpTrf proteins associate with phagocyte membranes through interactions with lipids, when rSpTrf-E1 is incubated with lipid-embedded nylon strips, it binds to phosphatidic acid (PA) through both the glycine-rich region and the histidine-rich region. Synthetic liposomes composed of PA and phosphatidylcholine show binding between rSpTrf-E1 and PA by fluorescence resonance energy transfer, which is associated with leakage of luminal contents suggesting changes in lipid organization and perhaps liposome lysis. Interactions with liposomes also change membrane curvature leading to liposome budding, fusion, and invagination, which is associated with PA clustering induced by rSpTrf-E1

  1. LPS-Induced Macrophage Activation and Plasma Membrane Fluidity Changes are Inhibited Under Oxidative Stress.

    Science.gov (United States)

    de la Haba, Carlos; Morros, Antoni; Martínez, Paz; Palacio, José R

    2016-12-01

    Macrophage activation is essential for a correct and efficient response of innate immunity. During oxidative stress membrane receptors and/or membrane lipid dynamics can be altered, leading to dysfunctional cell responses. Our aim is to analyze membrane fluidity modifications and cell function under oxidative stress in LPS-activated macrophages. Membrane fluidity of individual living THP-1 macrophages was evaluated by the technique two-photon microscopy. LPS-activated macrophage function was determined by TNFα secretion. It was shown that LPS activation causes fluidification of macrophage plasma membrane and production of TNFα. However, oxidative stress induces rigidification of macrophage plasma membrane and inhibition of cell activation, which is evidenced by a decrease of TNFα secretion. Thus, under oxidative conditions macrophage proinflammatory response might develop in an inefficient manner.

  2. Surface deformation of active volcanic areas retrieved with the SBAS-DInSAR technique: an overview

    Directory of Open Access Journals (Sweden)

    G. Zeni

    2008-06-01

    Full Text Available This paper presents a comprehensive overview of the surface deformation retrieval capability of the Differential Synthetic Aperture Radar Interferometry (DInSAR algorithm, referred to as Small BAseline Subset (SBAS technique, in the context of active volcanic areas. In particular, after a brief description of the algorithm some experiments relevant to three selected case-study areas are presented. First, we concentrate on the application of the SBAS algorithm to a single-orbit scenario, thus considering a set of SAR data composed by images acquired on descending orbits by the European Remote Sensing (ERS radar sensors and relevant to the Long Valley caldera (eastern California area. Subsequently, we address the capability of the SBAS technique in a multipleorbit context by referring to Mt. Etna volcano (southern Italy test site, with respect to which two different ERS data set, composed by images acquired both on ascending and descending orbits, are available. Finally, we take advantage of the capability of the algorithm to work in a multi-platform scenario by jointly exploiting two different sets of SAR images collected by the ERS and the Environment Satellite (ENVISAT radar sensors in the Campi Flegrei caldera (southern Italy area. The presented results demonstrate the effectiveness of the algorithm to investigate the deformation field in active volcanic areas and the potential of the DInSAR methodologies within routine surveillance scenario.

  3. Hydro-osmotic Instabilities in Active Membrane Tubes

    Science.gov (United States)

    Al-Izzi, Sami C.; Rowlands, George; Sens, Pierre; Turner, Matthew S.

    2018-03-01

    We study a membrane tube with unidirectional ion pumps driving an osmotic pressure difference. A pressure-driven peristaltic instability is identified, qualitatively distinct from similar tension-driven Rayleigh-type instabilities on membrane tubes. We discuss how this instability could be related to the function and biogenesis of membrane bound organelles, in particular, the contractile vacuole complex. The unusually long natural wavelength of this instability is in agreement with that observed in cells.

  4. Removal of Zn (II) and Ga (III) from waste waters using activated composite membranes

    International Nuclear Information System (INIS)

    Melita, L.; Meghea, A.; Munoz Tapia, M.; Gives, J. de

    2001-01-01

    The present study refers to the preparation of activated composite membrane (ACM) containing Aliquat 336 as a carrier, and testing their properties towards the selective transport of Ga and Zn cations. A new type of liquid membrane was prepared, named Activated Composite Membrane (ACM). The stability of these membrane increases, referring to other common membranes used before. These membranes have also good characteristics to separate metals. We cast membranes in two steps, first we used non-woven fabric (Hollytex 3329, France) as a support to manufacture reinforced polysulfone (PS) membrane which was obtained by the phase inversion technique, and second, a thin top layer of polyamide containing Aliquat 336 of two different concentrations (0.5 and 1 M) was obtained by interfacial polymerisation. The membrane thus prepared is composed of polyamide and polysulfone layers containing carrier. The surface texture of the membrane under study was examined by scanning electron microscopy (SEM) using a JSM-6300 scanning electron microscope. The chemical elemental analysis of freshly prepared membranes was performed, by X-ray diffraction measuring the energy distribution of the X-ray signal generated by a focused electron beam. A correlation between the carrier content in the membrane and the concentration of metal separated was obtained from the results of the membrane analysis by using the inductively coupled plasma (ICP) technique. The competition between gallium and zinc in the membrane surface is presented by the retaining membrane capacity. This type of membrane is relatively new for metal removal (Ga and Zn) from waste waters and the best cation retention was obtained for Zn. (authors)

  5. Purification of highly active alphavirus replication complexes demonstrates altered fractionation of multiple cellular membranes.

    Science.gov (United States)

    Pietilä, Maija K; van Hemert, Martijn J; Ahola, Tero

    2018-01-24

    Positive-strand RNA viruses replicate their genomes in membrane-associated structures; alphaviruses and many other groups induce membrane invaginations called spherules. Here, we established a protocol to purify these membranous replication complexes (RCs) from cells infected with Semliki Forest virus (SFV). We isolated SFV spherules located on the plasma membrane and further purified them using two consecutive density gradients. This revealed that SFV infection strongly modifies cellular membranes. We removed soluble proteins, the Golgi and most of the mitochondria, but plasma membrane, endoplasmic reticulum (ER) and late endosome markers enriched in the membrane fraction that contained viral RNA synthesizing activity, replicase proteins and minus- and plus-strand RNA. Electron microscopy revealed that the purified membranes displayed spherule-like structures with a narrow neck. This membrane enrichment was specific to viral replication as such a distribution of membrane markers was only observed after infection. Besides the plasma membrane, SFV infection remodeled the ER, and the co-fractionation of the RC-carrying plasma membrane and ER suggests that SFV may recruit ER proteins or membrane to the site of replication. The purified RCs were highly active in synthesizing both genomic and subgenomic RNA. Detergent solubilization destroyed the replication activity demonstrating that the membrane association of the complex is essential. Most of the newly made RNA was in double-stranded replicative molecules but the purified complexes also produced single-stranded RNA as well as released newly made RNA. This indicates that the purification established here maintained the functionality of RCs and thus enables further structural and functional studies of active RCs. IMPORTANCE Similar to all positive-strand RNA viruses, the arthropod-borne alphaviruses induce membranous genome factories but little is known about the arrangement of viral replicase proteins and the

  6. Plume Activity and Tidal Deformation on Enceladus Influenced by Faults and Variable Ice Shell Thickness.

    Science.gov (United States)

    Běhounková, Marie; Souček, Ondřej; Hron, Jaroslav; Čadek, Ondřej

    2017-09-01

    We investigated the effect of variations in ice shell thickness and of the tiger stripe fractures crossing Enceladus' south polar terrain on the moon's tidal deformation by performing finite element calculations in three-dimensional geometry. The combination of thinning in the polar region and the presence of faults has a synergistic effect that leads to an increase of both the displacement and stress in the south polar terrain by an order of magnitude compared to that of the traditional model with a uniform shell thickness and without faults. Assuming a simplified conductive heat transfer and neglecting the heat sources below the ice shell, we computed the global heat budget of the ice shell. For the inelastic properties of the shell described by a Maxwell viscoelastic model, we show that unrealistically low average viscosity of the order of 10 13 Pa s is necessary for preserving the volume of the ocean, suggesting the important role of the heat sources in the deep interior. Similarly, low viscosity is required to predict the observed delay of the plume activity, which hints at other delaying mechanisms than just the viscoelasticity of the ice shell. The presence of faults results in large spatial and temporal heterogeneity of geysering activity compared to the traditional models without faults. Our model contributes to understanding the physical mechanisms that control the fault activity, and it provides potentially useful information for future missions that will sample the plume for evidence of life. Key Words: Enceladus-Tidal deformation-Faults-Variable ice shell thickness-Tidal heating-Plume activity and timing. Astrobiology 17, 941-954.

  7. Active Deformation in the Overriding Plate Associated with Temporal Changes of the Philippine Sea Plate Motion

    Science.gov (United States)

    Ishiyama, T.; Sato, H.; Van Horne, A.

    2015-12-01

    We present detailed geologic evidence linking changes over time in Philippine Sea plate (PHS) motion and intracontinental deformation in central and southwest (SW) Japan during the Pliocene and after. In the early Pliocene, subduction of the PHS plate under SW Japan restarted in a northerly direction after period of deceleration or cessation. Later, motion changed to a more westerly direction. Corresponding geological changes found in the overriding plate include unconformities in the forearc basins, changes in slip sense on faults, depocenter migration, re-organization of drainage systems and volcanism. Quaternary intraplate deformation is prominent north of the Median Tectonic Line (MTL) inactive segment, above a shallow flat slab. In contrast, less Quaternary tectonic activity is found north of the MTL active segment which lies over a steadily-slipping portion of the subducting slab that behaves as a less-deformed rigid block. Depocenters and active thrusting have migrated north/northwestward over the past 5 My above the shallow flat slab segment of the PHS. We reconstructed the Plio-Pleistocene migration history using Neogene stratigraphy and shallow seismic reflection profiles. We see shallow PHS slab contact with the lower continental crust in our deep seismic reflection profiles, which may explain its enhanced downward drag of the overriding plate and synchronous strong compression in the crust. We find evidence of more westerly PHS plate subduction since the middle Pleistocene in (1) unconformities in the Kumano forearc basin deposits in SW Japan, (2) drastic stream captures in Shikoku, and (3) concordant changes in fault slip sense from thrust to dextral slip along the MTL. Oblique subduction could have induced stronger horizontal stress in the overriding plate above the shallow flat slab which could account for the increasing geologic slip rate observed on active structures. During four repetitions of megathrust earthquake sequences since the 17th century

  8. Detergent disruption of bacterial inner membranes and recovery of protein translocation activity

    International Nuclear Information System (INIS)

    Cunningham, K.; Wickner, W.T.

    1989-01-01

    Isolation of the integral membrane components of protein translocation requires methods for fractionation and functional reconstitution. The authors treated inner-membrane vesicles of Escherichia coli with mixtures of octyl β-D-glucoside, phospholipids, and an integral membrane carrier protein under conditions that extract most of the membrane proteins into micellar solution. Upon dialysis, proteoliposomes were reconstituted that supported translocation of radiochemically pure [ 35 S]pro-OmpA (the precursor of outer membrane protein A). Translocation into these proteoliposomes required ATP hydrolysis and membrane proteins, indicating that the reaction is that of the inner membrane. The suspension of membranes in detergent was separated into supernatant and pellet fractions by ultracentrifugation. After reconstitution, translocation activity was observed in both fractions, but processing by leader peptidase of translocated pro-OmpA to OmpA was not detectable in the reconstituted pellet fraction. Processing activity was restored by addition of pure leader peptidase as long as this enzyme was added before detergent removal, indicating that the translocation activity is not associated with detergent-resistant membrane vesicles. These results show that protein translocation activity can be recovered from detergent-disrupted membrane vesicles, providing a first step towards the goal of isolating the solubilized components

  9. Effect of crystal structure peculiarities of ageing alloys on dislocation structure during active deformation

    International Nuclear Information System (INIS)

    Travina, N.T.; Nikitin, A.A.; Zimina, L.N.

    1980-01-01

    On coarse-grain polycrystal samples of commercial KhN67MVTYu (EhP202), Kh60MVTYu (EhP487) and KhN60MKVYu (EhI661) alloys, aged under various conditions, studied are mechnanical properties after active deformation by extension at room temperature and dislocation structure corresponding to these structural states. Crystal and dislocation structures have been studied using electron microscope. In the alloys with mode--rate (13-14 v.%) quantity of γ' phase (KhN67MVTYu and KhN60MVTYu) at the stage of coherent-conjugated bond of particles with matrix deformation runs mainly according to the mechanism of cutting particles by separate dislocations; at the break of coherent bond - by bending of particles according to the Orovan mechanism or by their cutting with clusters of dislocations. At high volume part of particles of γ'-phase (40 v%, KhN60MKVYu alloy) the Orovan mechanism does not manifest itself in any of studied states. In this alloy at this stage of the break of the γ'-phase coherent bond with matrix in the result of ordered position of particles along the directions in matrix formed are interlayers free from particles on which strain is being mainly developed. Dislocation structure is compared with mechanical properties of the alloys

  10. Oxygen activation at the plasma membrane: relation between superoxide and hydroxyl radical production by isolated membranes.

    Science.gov (United States)

    Heyno, Eiri; Mary, Véronique; Schopfer, Peter; Krieger-Liszkay, Anja

    2011-07-01

    Production of reactive oxygen species (hydroxyl radicals, superoxide radicals and hydrogen peroxide) was studied using EPR spin-trapping techniques and specific dyes in isolated plasma membranes from the growing and the non-growing zones of hypocotyls and roots of etiolated soybean seedlings as well as coleoptiles and roots of etiolated maize seedlings. NAD(P)H mediated the production of superoxide in all plasma membrane samples. Hydroxyl radicals were only produced by the membranes of the hypocotyl growing zone when a Fenton catalyst (FeEDTA) was present. By contrast, in membranes from other parts of the seedlings a low rate of spontaneous hydroxyl radical formation was observed due to the presence of small amounts of tightly bound peroxidase. It is concluded that apoplastic hydroxyl radical generation depends fully, or for the most part, on peroxidase localized in the cell wall. In soybean plasma membranes from the growing zone of the hypocotyl pharmacological tests showed that the superoxide production could potentially be attributed to the action of at least two enzymes, an NADPH oxidase and, in the presence of menadione, a quinone reductase.

  11. Estimating Actual 2D Ground Deformations Induced by Underground Activities with Cross-Heading InSAR Measurements

    Directory of Open Access Journals (Sweden)

    Xiaoying Yu

    2017-01-01

    Full Text Available InSAR can only monitor relative ground deformations with respect to a reference area. In order to obtain actual deformations, GCPs or stable area is required in the study area, which, however, may be unavailable in the investigating of geohazards associated with underground activities (i.e., groundwater pumping, underground mining, and oil/gas exploitation. We propose a novel approach to estimate actual 2D deformations based on the InSAR relative LOS measurements acquired from cross-heading datasets. The errors induced by the arbitrary selection of reference areas can thus be avoided. The performance of the proposed approach is validated by a series of simulations. By providing the ascending and descending measurements with errors of 2 and 1.5 mm/year STDs, respectively, the RMSEs are 2.1 and 2.6 mm/year for the estimated vertical and east deformations, respectively. A case study is carried out in Cangzhou, China, for estimating the actual 2D ground deformations associated with groundwater pumping. By integrating ALOS ascending and ENVISAT descending datasets acquired between 2007 and 2010, we found that the Cangzhou area experienced ground subsidence of up to 23.4 mm/year in the suburbs but ground uplift of up to 20.9 mm/year in the urban area, both of which are accompanied by considerable lateral deformations.

  12. Tissue Deformation Index as a Reliable Measure of Lateral Abdominal Muscle Activation on M-Mode Sonography.

    Science.gov (United States)

    Biały, Maciej; Adamczyk, Wacław; Gnat, Rafael; Stranc, Tomasz

    2017-07-01

    The aim of this article is to present a novel method of evaluating the activity of lateral abdominal muscles using M-mode sonography. The method leads to calculation of the tissue deformation index, representing the percent change in lateral abdominal muscle thickness over time. The objectives of this study were as follows: (1) to establish the mean tissue deformation index values for individual lateral abdominal muscles; and (2) to establish the reliability of the tissue deformation index. In a group of 34 healthy young volunteers (mean age, 24.03 years; body mass, 68.89 kg; body height, 174.25 cm), the reflex response of the lateral abdominal muscles to postural perturbation in the form of rapid arm abduction was recorded in 2 series, with 6 repetitions each, and the tissue deformation index was calculated. The mean tissue deformation index values formed a gradient, increasing from deep to superficial lateral abdominal muscles: 0.06%/ms for transversus abdominis, 0.11%/ms for oblique internal, and 0.16 for oblique external muscles. The tissue deformation index values differed significantly among individual lateral abdominal muscles (all paired comparisons, P  0.8). © 2017 by the American Institute of Ultrasound in Medicine.

  13. Analysis of mitochondrial 3D-deformation in cardiomyocytes during active contraction reveals passive structural anisotropy of orthogonal short axes.

    Directory of Open Access Journals (Sweden)

    Yael Yaniv

    Full Text Available The cardiomyocyte cytoskeleton, composed of rigid and elastic elements, maintains the isolated cell in an elongated cylindrical shape with an elliptical cross-section, even during contraction-relaxation cycles. Cardiomyocyte mitochondria are micron-sized, fluid-filled passive spheres distributed throughout the cell in a crystal-like lattice, arranged in pairs sandwiched between the sarcomere contractile machinery, both longitudinally and radially. Their shape represents the extant 3-dimensional (3D force-balance. We developed a novel method to examine mitochondrial 3D-deformation in response to contraction and relaxation to understand how dynamic forces are balanced inside cardiomyocytes. The variation in transmitted light intensity induced by the periodic lattice of myofilaments alternating with mitochondrial rows can be analyzed by Fourier transformation along a given cardiomyocyte axis to measure mitochondrial deformation along that axis. This technique enables precise detection of changes in dimension of ∼1% in ∼1 µm (long-axis structures with 8 ms time-resolution. During active contraction (1 Hz stimulation, mitochondria deform along the length- and width-axes of the cell with similar deformation kinetics in both sarcomere and mitochondrial structures. However, significant deformation anisotropy (without hysteresis was observed between the orthogonal short-axes (i.e., width and depth of mitochondria during electrical stimulation. The same degree of deformation anisotropy was also found between the myocyte orthogonal short-axes during electrical stimulation. Therefore, the deformation of the mitochondria reflects the overall deformation of the cell, and the apparent stiffness and stress/strain characteristics of the cytoskeleton differ appreciably between the two cardiomyocyte orthogonal short-axes. This method may be applied to obtaining a better understanding of the dynamic force-balance inside cardiomyocytes and of changes in the

  14. Chitosan/Carboxymethylcellulose/Ionic Liquid/Ag(0 Nanoparticles Form a Membrane with Antimicrobial Activity

    Directory of Open Access Journals (Sweden)

    Camila Quadros

    2013-01-01

    Full Text Available Silver metal nanoparticles were immobilized in chitosan/carboxymethylcellulose/BMI.BF4(1-n-butyl-3-methylimidazolium tetrafluoroborate ionic liquid (CS/CMC/IL to form polymeric membrane with 20 μm thickness. The CS/CMC/IL polymeric membrane was prepared using a simple solution blending method. Irregularly shaped Ag(0 nanoparticles with monomodal size distributions of 8.0±0.4 nm Ag(0 were immobilized in the membrane. The presence of small Ag(0 nanoparticles induced an augmentation in the CS/CMC/IL film surface areas. The CS/CMC/IL membrane containing Ag(0 showed increase antimicrobial activity the Ag(0 concentration increased up to saturation at 10 mg. CS/CMC/IL membrane that contains Ag(0 nanoparticles has enhanced durability of the membrane and exhibited stronger antimicrobial activity against Escherichia coli and Staphylococcus aureus.

  15. Influence of the lipid membrane environment on structure and activity of the outer membrane protein Ail from Yersinia pestis.

    Science.gov (United States)

    Ding, Yi; Fujimoto, L Miya; Yao, Yong; Plano, Gregory V; Marassi, Francesca M

    2015-02-01

    The surrounding environment has significant consequences for the structural and functional properties of membrane proteins. While native structure and function can be reconstituted in lipid bilayer membranes, the detergents used for protein solubilization are not always compatible with biological activity and, hence, not always appropriate for direct detection of ligand binding by NMR spectroscopy. Here we describe how the sample environment affects the activity of the outer membrane protein Ail (attachment invasion locus) from Yersinia pestis. Although Ail adopts the correct β-barrel fold in micelles, the high detergent concentrations required for NMR structural studies are not compatible with the ligand binding functionality of the protein. We also describe preparations of Ail embedded in phospholipid bilayer nanodiscs, optimized for NMR studies and ligand binding activity assays. Ail in nanodiscs is capable of binding its human ligand fibronectin and also yields high quality NMR spectra that reflect the proper fold. Binding activity assays, developed to be performed directly with the NMR samples, show that ligand binding involves the extracellular loops of Ail. The data show that even when detergent micelles support the protein fold, detergents can interfere with activity in subtle ways. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. Post-activation depression of the lower extremities in stroke patients with spasticity and spastic equinovarus deformity

    Directory of Open Access Journals (Sweden)

    Yuanbin Yang

    2015-06-01

    Full Text Available This study aimed to investigate changes of post-activation depression in two groups of patients with or without spastic equinovarus deformity (SED. Paired and independent t-tests were used to compare post-activation depression within and between the groups, respectively. There was a significant positive correlation between diminished post-activation depression and spasticity severity. The soleus and tibialis anterior (TA post-activation depression values were significantly decreased on the affected sides of patients with SED compared to those without. In patients without SED, the soleus post-activation depression was significantly decreased on the affected side; however, TA post-activation depression was higher on the affected side. Both the soleus and TA become active, but the onset time may be different. The imbalanced muscle tone between the soleus and TA in the early stage after stroke may be related to equinus deformity.

  17. Active deformation processes of the Northern Caucasus deduced from the GPS observations

    Science.gov (United States)

    Milyukov, Vadim; Mironov, Alexey; Rogozhin, Eugeny; Steblov, Grigory; Gabsatarov, Yury

    2015-04-01

    The Northern Caucasus, as a part of the Alpine-Himalayan mobile belt, is a zone of complex tectonics associated with the interaction of the two major tectonic plates, Arabian and Eurasian. The first GPS study of the contemporary geodynamics of the Caucasus mountain system were launched in the early 1990s in the framework of the Russia-US joint project. Since 2005 observations of the modern tectonic motion of the Northern Caucasus are carried out using the continuous GPS network. This network encompasses the territory of three Northern Caucasian Republics of the Russian Federation: Karachay-Cherkessia, Kabardino-Balkaria, and North Ossetia. In the Ossetian part of the Northern Caucasus the network of GPS survey-mode sites has been deployed as well. The GPS velocities confirm weak general compression of the Northern Caucasus with at the rate of about 1-2 mm/year. This horizontal motion at the boundary of the Northern Caucasus with respect to the Eurasian plate causes the higher seismic and tectonic activity of this transition zone. This result confirms that the source of deformation of the Northern Caucasus is the sub-meridional drift of the Arabian plate towards the adjacent boundary of the Eastern European part of the Eurasian lithospheric plate. The concept of such convergence implies that the Caucasian segment of the Alpine-Himalayan mobile belt is under compression, the layers of sedimentary and volcanic rocks are folded, the basement blocks are subject to shifts in various directions, and the upper crust layers are ruptured by reverse faults and thrusts. Weak deviation of observed velocities from the pattern corresponding to homogeneous compression can also be revealed, and numerical modeling of deformations of major regional tectonic structures, such as the Main Caucasus Ridge, can explain this. The deformation tensor deduced from the velocity field also exhibits the sub-meridional direction of the major compressional axes which coincides with the direction of

  18. A coupled theory for chemically active and deformable solids with mass diffusion and heat conduction

    Science.gov (United States)

    Zhang, Xiaolong; Zhong, Zheng

    2017-10-01

    To analyse the frequently encountered thermo-chemo-mechanical problems in chemically active material applications, we develop a thermodynamically-consistent continuum theory of coupled deformation, mass diffusion, heat conduction and chemical reaction. Basic balance equations of force, mass and energy are presented at first, and then fully coupled constitutive laws interpreting multi-field interactions and evolving equations governing irreversible fluxes are constructed according to the energy dissipation inequality and the chemical kinetics. To consider the essential distinction between mass diffusion and chemical reactions in affecting free energy and dissipations of a highly coupled system, we regard both the concentrations of diffusive species and the extent of reaction as independent state variables. This new formulation then distinguishes between the energy contribution from the diffusive species entering the solid and that from the subsequent chemical reactions occurring among these species and the host solid, which not only interact with stresses or strains in different manners and on different time scales, but also induce different variations of solid microstructures and material properties. Taking advantage of this new description, we further establish a specialized isothermal model to predict precisely the transient chemo-mechanical response of a swelling solid with a proposed volumetric constraint that accounts for material incompressibility. Coupled kinetics is incorporated to capture the volumetric swelling of the solid caused by imbibition of external species and the simultaneous dilation arised from chemical reactions between the diffusing species and the solid. The model is then exemplified with two numerical examples of transient swelling accompanied by chemical reaction. Various ratios of characteristic times of diffusion and chemical reaction are taken into account to shed light on the dependency on kinetic time scales of evolution patterns for

  19. A Catalytically Active Membrane Reactor for Fast, Highly Exothermic, Heterogeneous Gas Reactions. A Pilot Plant Study

    NARCIS (Netherlands)

    Veldsink, Jan W.; Versteeg, Geert F.; Swaaij, Wim P.M. van

    1995-01-01

    Membrane reactors have been frequently studied because of their ability to combine chemical activity and separation properties into one device. Due to their thermal stability and mechanical strength, ceramic membranes are preferred over polymeric ones, but small transmembrane fluxes obstruct a

  20. The influence of activation of heterogeneous ion-exchange membranes on their electrochemical properties

    Czech Academy of Sciences Publication Activity Database

    Brožová, Libuše; Křivčík, J.; Neděla, D.; Kysela, V.; Žitka, Jan

    2015-01-01

    Roč. 56, č. 12 (2015), s. 3228-3232 ISSN 1944-3994. [International Conference on Membrane and Electromembrane Processes - MELPRO 2014. Prague, 18.05.2014-21.05.2014] Institutional support: RVO:61389013 Keywords : heterogeneous ion-exchange membranes * electrochemical properties * activation Subject RIV: JP - Industrial Processing Impact factor: 1.272, year: 2015

  1. Why low powdered activated carbon addition reduces membrane fouling in MBRs

    NARCIS (Netherlands)

    Remy, M.J.J.; Potier, V.; Temmink, B.G.; Rulkens, W.H.

    2010-01-01

    Previous research had demonstrated that powdered activated carbon (PAC), when applied at very low dosages and long SRTs, reduces membrane fouling in membrane bioreactor (MBRs). In this contribution several mechanisms to explain this beneficial effect of PAC were investigated, including enhanced

  2. Preparation, Characterization and Performance Studies of Active PVDF Ultrafiltration-Surfactants Membranes Containing PVP as Additive

    International Nuclear Information System (INIS)

    Nur Izzah Md Fadilah; Abdul Rahman Hassan

    2016-01-01

    The role of surfactants in the formation of active Poly(vinylidene fluoride) (PVDF) ultrafiltration (AUF) membranes was studied. The effect combination of surfactants that are Sodium dodecyl sulfate (SDS)/ Tween 80 and Tween 80/ Triton X-100 formulations on performance and morphological structures were investigated for the first time. The influence of surfactants blends on the membrane pores was also examined. Experimental data showed that combination of Tween 80/ Triton X-100 give the highest BSA permeation flux with a value of 285.51 Lm -2 h -1 . With combination of SDS/ Tween 80, the AUF membrane showed the highest protein rejection up to 93 % and 79 % for Bovine Serum Albumin (BSA) and Egg Albumin (EA), respectively. Moreover, membranes characterization demonstrated that the addition of SDS/ Tween 80 and Tween 80/ Triton X-100 were found to affect the performance, surface morphologies and membrane pores of AUF PVDF membranes. (author)

  3. Active endocannabinoids are secreted on extracellular membrane vesicles.

    Science.gov (United States)

    Gabrielli, Martina; Battista, Natalia; Riganti, Loredana; Prada, Ilaria; Antonucci, Flavia; Cantone, Laura; Matteoli, Michela; Maccarrone, Mauro; Verderio, Claudia

    2015-02-01

    Endocannabinoids primarily influence neuronal synaptic communication within the nervous system. To exert their function, endocannabinoids need to travel across the intercellular space. However, how hydrophobic endocannabinoids cross cell membranes and move extracellularly remains an unresolved problem. Here, we show that endocannabinoids are secreted through extracellular membrane vesicles produced by microglial cells. We demonstrate that microglial extracellular vesicles carry on their surface N-arachidonoylethanolamine (AEA), which is able to stimulate type-1 cannabinoid receptors (CB1), and inhibit presynaptic transmission, in target GABAergic neurons. This is the first demonstration of a functional role of extracellular vesicular transport of endocannabinoids. © 2015 The Authors.

  4. G-protein activity in Percoll-purified plasma membranes, bulk plasma membranes, and low-density plasma membranes isolated from rat cerebral cortex

    Czech Academy of Sciences Publication Activity Database

    Bouřová, Lenka; Stöhr, Jiří; Lisý, Václav; Rudajev, Vladimír; Novotný, Jiří; Svoboda, Petr

    2009-01-01

    Roč. 15, č. 4 (2009), BR111-BR122 ISSN 1234-1010 R&D Projects: GA MŠk(CZ) LC554; GA MŠk(CZ) LC06063; GA ČR(CZ) GA309/06/0121; GA AV ČR(CZ) IAA500110606 Institutional research plan: CEZ:AV0Z50110509 Keywords : rat cerebral cortex * plasma membrane * G-protein activity Subject RIV: CE - Biochemistry Impact factor: 1.543, year: 2009

  5. Active faulting on the Ninetyeast Ridge and its relation to deformation of the Indo-Australian plate

    Digital Repository Service at National Institute of Oceanography (India)

    Sager, W.W.; Bull, J.M.; Krishna, K.S.

    extents are poorly defined. New multichannel seismic reflection profiles image active faults along the entire length of the NER and show spatial changes in the style of deformation along the ridge. The northern NER (0°N–5°N) displays transpressional motion...

  6. Chimaerin suppresses Rac1 activation at the apical membrane to maintain the cyst structure.

    Directory of Open Access Journals (Sweden)

    Shunsuke Yagi

    Full Text Available Epithelial organs are made of a well-polarized monolayer of epithelial cells, and their morphology is maintained strictly for their proper functions. Previously, we showed that Rac1 activation is suppressed at the apical membrane in the mature organoid, and that such spatially biased Rac1 activity is required for the polarity maintenance. Here we identify Chimaerin, a GTPase activating protein for Rac1, as a suppressor of Rac1 activity at the apical membrane. Depletion of Chimaerin causes over-activation of Rac1 at the apical membrane in the presence of hepatocyte growth factor (HGF, followed by luminal cell accumulation. Importantly, Chimaerin depletion did not inhibit extension formation at the basal membrane. These observations suggest that Chimaerin functions as the apical-specific Rac1 GAP to maintain epithelial morphology.

  7. Guanidination of notexin alters its membrane-damaging activity in ...

    Indian Academy of Sciences (India)

    MADHU

    Wilson H I and Nicholson G M 1997 Presynaptic snake beta- neurotoxins produce tetanic fade and endplate potential run- down during neuromuscular blockade in mouse diaphragm;. Naunyn Schmiedebergs Arch. Pharmacol. 356 626–634. Xu X and London E 2000 The effect of sterol structure on membrane lipid domains ...

  8. In vitro antioxidant and membrane stabilization activities of the fruit ...

    African Journals Online (AJOL)

    Arthritis is an inflammation of one or more joints. There is considerable experimental evidence linking lysosomal enzymes with tissue damage in arthritis. This study investigated anti-arthritic properties of Tetrapleura tetraptera (TT) using membrane stabilization assay (MSA). Powdered TT fruit sample was extracted by ...

  9. The roles of membrane microdomains (rafts) in T cell activation

    Czech Academy of Sciences Publication Activity Database

    Hořejší, Václav

    2003-01-01

    Roč. 191, - (2003), s. 148-164 ISSN 0105-2896 R&D Projects: GA MŠk LN00A026 Grant - others:Wellcome Trust(GB) J1116W24Z Institutional research plan: CEZ:AV0Z5052915 Keywords : membrane microdomain * raft * T cell Subject RIV: EC - Immunology Impact factor: 7.052, year: 2003

  10. Designing CNC Knit for Hybrid Membrane And Bending Active Structures

    DEFF Research Database (Denmark)

    Tamke, Martin; Holden Deleuran, Anders; Gengnagel, Christoph

    2015-01-01

    specific properties and detailing. CNC knitting with high tenacity yarn enables this practice and offers an alternative to current woven membranes. The design and fabrication of an 8m high fabric tower through an interdisciplinary team of architects, structural and textile engineers, allowed to investigate...

  11. Translocation across biological membranes: activity, structure and regulation of transporters

    NARCIS (Netherlands)

    Ruiz, Stephanie

    2017-01-01

    Alle cellen zijn omsloten door biologische membranen met selectief doorlatende membrane, die het mogelijk maken om omstandigheden "binnen" en "buiten" de cel te handhaven. De gereguleerde beweging van moleculen door deze membranen is nodig voor talrijke essentiële processen, waaronder het verkrijgen

  12. Cellulose microfibril deposition: coordinated activity at the plant plasma membrane

    NARCIS (Netherlands)

    Lindeboom, J.J.; Mulder, B.; Vos, J.W.; Ketelaar, M.J.; Emons, A.M.C.

    2008-01-01

    Plant cell wall production is a membrane-bound process. Cell walls are composed of cellulose microfibrils, embedded inside a matrix of other polysaccharides and glycoproteins. The cell wall matrix is extruded into the existing cell wall by exocytosis. This same process also inserts the cellulose

  13. Tectonic and gravity-induced deformation along the active Talas-Fergana Fault, Tien Shan, Kyrgyzstan

    Science.gov (United States)

    Tibaldi, A.; Corazzato, C.; Rust, D.; Bonali, F. L.; Pasquarè Mariotto, F. A.; Korzhenkov, A. M.; Oppizzi, P.; Bonzanigo, L.

    2015-08-01

    This paper shows, by field palaeoseismological data, the Holocene activity of the central segment of the intracontinental Talas-Fergana Fault (TFF), and the relevance of possible future seismic shaking on slope stability around a large water reservoir. The fault, striking NW-SE, is marked by a continuous series of scarps, deflected streams and water divides, and prehistoric earthquakes that offset substrate and Holocene deposits. Fault movements are characterised by right-lateral strike-slip kinematics with a subordinate component of uplift of the NE block. Structural, geological and geomorphological field data indicate that shallow and deep landslides are aligned along the TFF, and some of them are active. Where the TFF runs close to the reservoir, the fault trace is obscured by a series of landslides, affecting rock and soil materials and ranging in size from small slope instabilities to deep-seated gravity-induced slope deformations (DGSDs). The largest of these, which does not show clear evidence of present-day activity, involves a volume of about 1 km3 and is associated with smaller but active landslides in its lower part, with volumes in the order of 2.5 × 104 m3 to 1 × 106 m3. Based on the spatial and temporal relations between landslides and faults, we argue that at least some of these slope failures may have a coseismic character. Stability analyses by means of limit equilibrium methods (LEMs), and stress-strain analysis by finite difference numerical modelling (FDM), were carried out to evaluate different hazard scenarios linked to these slope instabilities. The results indicate concern for the different threats posed, ranging from the possible disruption of the M-41 highway, the main transportation route in central Asia, to the possible collapse of huge rock masses into the reservoir, possibly generating a tsunami.

  14. Killing of Staphylococci by θ-Defensins Involves Membrane Impairment and Activation of Autolytic Enzymes

    Directory of Open Access Journals (Sweden)

    Miriam Wilmes

    2014-11-01

    Full Text Available θ-Defensins are cyclic antimicrobial peptides expressed in leukocytes of Old world monkeys. To get insight into their antibacterial mode of action, we studied the activity of RTDs (rhesus macaque θ-defensins against staphylococci. We found that in contrast to other defensins, RTDs do not interfere with peptidoglycan biosynthesis, but rather induce bacterial lysis in staphylococci by interaction with the bacterial membrane and/or release of cell wall lytic enzymes. Potassium efflux experiments and membrane potential measurements revealed that the membrane impairment by RTDs strongly depends on the energization of the membrane. In addition, RTD treatment caused the release of Atl-derived cell wall lytic enzymes probably by interaction with membrane-bound lipoteichoic acid. Thus, the premature and uncontrolled activity of these enzymes contributes strongly to the overall killing by θ-defensins. Interestingly, a similar mode of action has been described for Pep5, an antimicrobial peptide of bacterial origin.

  15. Enhancement of antibacterial activity in nanofillers incorporated PSF/PVP membranes

    Science.gov (United States)

    Pramila, P.; Gopalakrishnan, N.

    2018-04-01

    An attempt has been made to investigate the nanofillers incorporated polysulfone (PSF) and polyvinylpyrrolidone (PVP) polymer membranes prepared by phase inversion method. Initially, the nanofillers, viz, Zinc Oxide (ZnO) nanoparticle, Graphene Oxide-Zinc Oxide (GO-ZnO) nanocomposite were synthesized and then directly incorporated into PSF/PVP blend during the preparation of membranes. The prepared membranes have been subjected to FE-SEM, AFM, BET, contact angle, tensile test and anti-bacterial studies. Significant membrane morphologies and nanoporous properties have been observed by FE-SEM and BET, respectively. It has been observed that hydrophilicity, mechanical strength and water permeability of the ZnO and GO-ZnO incorporated membranes were enhanced than bare membrane. Antibacterial activity was assessed by measuring the inhibition zones formed around the membrane by disc-diffusion method using Escherichia coli (gram-negative) as a model bacterium. Again, it has been observed that nanofillers incorporated membrane exhibits high antibacterial performance compared to bare membrane.

  16. Modification of trout sperm membranes associated with activation and cryopreservation. Implications for fertilizing potential

    Science.gov (United States)

    Abstract We investigated the effects of two trout sperm activation solutions on sperm physiology and membrane organization prior to and following cryopreservation using flow cytometry and investigated their impact on in vitro fertility. Cryopreservation caused greater phospholipid disorder (high pl...

  17. Machine learning-enabled discovery and design of membrane-active peptides.

    Science.gov (United States)

    Lee, Ernest Y; Wong, Gerard C L; Ferguson, Andrew L

    2017-07-08

    Antimicrobial peptides are a class of membrane-active peptides that form a critical component of innate host immunity and possess a diversity of sequence and structure. Machine learning approaches have been profitably employed to efficiently screen sequence space and guide experiment towards promising candidates with high putative activity. In this mini-review, we provide an introduction to antimicrobial peptides and summarize recent advances in machine learning-enabled antimicrobial peptide discovery and design with a focus on a recent work Lee et al. Proc. Natl. Acad. Sci. USA 2016;113(48):13588-13593. This study reports the development of a support vector machine classifier to aid in the design of membrane active peptides. We use this model to discover membrane activity as a multiplexed function in diverse peptide families and provide interpretable understanding of the physicochemical properties and mechanisms governing membrane activity. Experimental validation of the classifier reveals it to have learned membrane activity as a unifying signature of antimicrobial peptides with diverse modes of action. Some of the discriminating rules by which it performs classification are in line with existing "human learned" understanding, but it also unveils new previously unknown determinants and multidimensional couplings governing membrane activity. Integrating machine learning with targeted experimentation can guide both antimicrobial peptide discovery and design and new understanding of the properties and mechanisms underpinning their modes of action. Copyright © 2017 Elsevier Ltd. All rights reserved.

  18. Hydrophilic Phage-Mimicking Membrane Active Antimicrobials Reveal Nanostructure-Dependent Activity and Selectivity.

    Science.gov (United States)

    Jiang, Yunjiang; Zheng, Wan; Kuang, Liangju; Ma, Hairong; Liang, Hongjun

    2017-09-08

    The prevalent wisdom on developing membrane active antimicrobials (MAAs) is to seek a delicate, yet unquantified, cationic-hydrophobic balance. Inspired by phages that use nanostructured protein devices to invade bacteria efficiently and selectively, we study here the antibiotic role of nanostructures by designing spherical and rod-like polymer molecular brushes (PMBs) that mimic the two basic structural motifs of bacteriophages. Three model PMBs with different well-defined geometries consisting of multiple, identical copies of densely packed poly(4-vinyl-N-methylpyridine iodide) branches are synthesized by controlled/"living" polymerization, reminiscent of the viral structural motifs comprised of multiple copies of protein subunits. We show that, while the individual linear-chain polymer branch that makes up the PMBs is hydrophilic and a weak antimicrobial, amphiphilicity is not a required antibiotic trait once nanostructures come into play. The nanostructured PMBs induce an unusual topological transition of bacterial but not mammalian membranes to form pores. The sizes and shapes of the nanostructures further help define the antibiotic activity and selectivity of the PMBs against different families of bacteria. This study highlights the importance of nanostructures in the design of MAAs with high activity, low toxicity, and target specificity.

  19. The Effects of Altered Membrane Cholesterol Levels on Sodium Pump Activity in Subclinical Hypothyroidism

    Directory of Open Access Journals (Sweden)

    Suparna Roy

    2017-02-01

    Full Text Available BackgroundMetabolic dysfunctions characteristic of overt hypothyroidism (OH start at the early stage of subclinical hypothyroidism (SCH. Na+/K+-ATPase (the sodium pump is a transmembrane enzyme that plays a vital role in cellular activities in combination with membrane lipids. We evaluated the effects of early changes in thyroid hormone and membrane cholesterol on sodium pump activity in SCH and OH patients.MethodsIn 32 SCH patients, 35 OH patients, and 34 euthyroid patients, sodium pump activity and cholesterol levels in red blood cell membranes were measured. Serum thyroxine (T4 and thyroid stimulating hormone (TSH levels were measured using enzyme-linked immunosorbent assays. Differences in their mean values were analysed using post hoc analysis of variance. We assessed the dependence of the sodium pump on other metabolites by multiple regression analysis.ResultsSodium pump activity and membrane cholesterol were lower in both hypothyroid groups than in control group, OH group exhibiting lower values than SCH group. In SCH group, sodium pump activity showed a significant direct dependence on membrane cholesterol with an inverse relationship with serum TSH levels. In OH group, sodium pump activity depended directly on membrane cholesterol and serum T4 levels. No dependence on serum cholesterol was observed in either case.ConclusionDespite the presence of elevated serum cholesterol in hypothyroidism, membrane cholesterol contributed significantly to maintain sodium pump activity in the cells. A critical reduction in membrane cholesterol levels heralds compromised enzyme activity, even in the early stage of hypothyroidism, and this can be predicted by elevated TSH levels alone, without any evident clinical manifestations.

  20. On Loosening Plastic Composite under Active Load and Its Influence on the Deformation and Strength Properties

    Directory of Open Access Journals (Sweden)

    K. F. Komkov

    2015-01-01

    Full Text Available Processing the test results of the composite, which is a mechanical mixture of metal particles with a plastic polymer binder, has shown that its deformation and strength properties are substantially different from those of stable plastic material. The specimen tests for tensile and compression with measuring transverse deformations, as well as torsion tests of tubular samples have revealed that the process of its deformation is accompanied by a change in the original structure.The composite instability is caused by the fact that during this process, it acquires considerable loosening that depends on the type of the stress-state. Hard metal particles are hardly deformed at any stress-state, but they form a layer of bonds that affect the mixture behavior under force action. The total deformation is the plastic flow of the binder on which deformation, caused both by sliding and by loss of the surface layer bonds, is superimposed.The analysis shows that with destruction at tensile test the non-linear part of the bulk deformation (dilatancy is 6 times more than "conditionally" elastic (3.5 times compressed. The objective of this work is to develop a technique for determining a dilatancy, define its influence on deformation and strength properties of the composite, and improve the mathematical model of the material. The proposed model based on the tensor-nonlinear equations describes loosening, as an additional component of the mean deformation and as a mean stress component, hereinafter referred to as: the first - by the deformation, the second – by the stress. A ratio value of the nonlinear part of deformation with the quadratic tensor argument to the linear part, which reaches 0.3, shows the need for such equations. It also shows the influence of deformation on the relationship between the deviators.To enhance capabilities of mathematical model is possible after including therein the equations for the spherical part of the tensor of deformation

  1. Effect of activated carbon in PSF-PEI-Ag symmetric membrane

    Directory of Open Access Journals (Sweden)

    Said Khairul Anwar Mohamad

    2017-01-01

    Full Text Available Polysulfone (Psf composite membrane consist of activated carbon, polyethyleneimine and silver nitrate was prepared by phase inversion. The activated carbon (AC act as adsorbent to adsorb heavy metal present in synthetic waste water while polysulfone membrane act as support. Phase inversion was carried out on different composition of activated carbon from 0 to 0.9% while other component are remain constant. The surface morphology of composite membrane was characterized by scanning electron microscopy (SEM while heavy metal absorption was quantified by atomic absorption spectrometer (AAS. The SEM image show symmetric membrane matrix with sponge structure. The composite membrane with 0.9wt% AC has the highest water flux as well as removal of heavy metal (chromium, lead, silver and cadmium compare to composite membrane with 0.3wt% AC and 0.5wt% AC. The percentage of heavy metal reduction by composite membrane 0.9wt% AC was 35% cadmium, 19% chromium, 16% silver and 2% lead. The result indicated that the introduction of 0.9wt% AC indeed plays an important role towards enhancing the adsorption of heavy metal in water.

  2. The Bile Acid-Sensitive Ion Channel (BASIC) Is Activated by Alterations of Its Membrane Environment

    Science.gov (United States)

    Schmidt, Axel; Lenzig, Pia; Oslender-Bujotzek, Adrienne; Kusch, Jana; Dias Lucas, Susana; Gründer, Stefan; Wiemuth, Dominik

    2014-01-01

    The bile acid-sensitive ion channel (BASIC) is a member of the DEG/ENaC family of ion channels. Channels of this family are characterized by a common structure, their physiological functions and modes of activation, however, are diverse. Rat BASIC is expressed in brain, liver and intestinal tract and activated by bile acids. The physiological function of BASIC and its mechanism of bile acid activation remain a puzzle. Here we addressed the question whether amphiphilic bile acids activate BASIC by directly binding to the channel or indirectly by altering the properties of the surrounding membrane. We show that membrane-active substances other than bile acids also affect the activity of BASIC and that activation by bile acids and other membrane-active substances is non-additive, suggesting that BASIC is sensitive for changes in its membrane environment. Furthermore based on results from chimeras between BASIC and ASIC1a, we show that the extracellular and the transmembrane domains are important for membrane sensitivity. PMID:25360526

  3. Structural deformation and sedimentation in an active Caldera, Rabaul, Papua New Guinea

    Science.gov (United States)

    Greene, H. Gary; Tiffin, D.L.; McKee, C.O.

    1986-01-01

    Recent seismic and tectonic activity in Rabaul Caldera, Papua New Guinea, suggests that magma is accumulating at a shallow depth beneath this partially submerged structure and that a new volcano may be developing. Changes in onshore elevation since 1971 (as much as 2 m on south Matupit Island) indicate that rapid and large-scale uplifts have occurred on the seafloor near the center of the caldera. The frequency of seismic events within the caldera has also increased during this period. Earthquake locations define an elliptical ring surrounding the center of this uplift within the caldera. A marine geophysical survey in 1982 by the U.S. Geological Survey's R/V "S.P. Lee" in Rabaul Caldera shows the development of a bulge in the seafloor near the center of the caldera. High-resolution seismic reflection profiles show that this bulge consists of two domal uplifts bounded and separated by two major north-south-trending fault zones. Deformed sediments overlie these zones; a prominent slump flanks the area of the bulge. Five major acoustic units were identified in the seismic reflection profiles: an acoustic basement and four sedimentary units consisting of irregularly layered, cross-layered, contorted, and well-layered sequences. The acoustic basement is probably composed of crystalline volcanic rocks, and the layered acoustic units are probably sediments, primarily ash deposited in different environments. The cross-layered, irregularly layered, and contorted units appear to have been deposited in a dynamic environment subjected to strong currents, seismicity, and/or mass wasting, while the well-layered units were deposited in a low-energy environment. Locally, well-layered sequences interfinger with the other sedimentary units, indicating a transitional environment that alternated between high-energy and low-energy depositional processes. A submarine channel cuts most of the acoustic units and appears to be the conduit for sediment transport out of the caldera; it

  4. The higher level of complexity of K-Ras4B activation at the membrane

    Science.gov (United States)

    Jang, Hyunbum; Banerjee, Avik; Chavan, Tanmay S.; Lu, Shaoyong; Zhang, Jian; Gaponenko, Vadim; Nussinov, Ruth

    2016-01-01

    Is nucleotide exchange sufficient to activate K-Ras4B? To signal, oncogenic rat sarcoma (Ras) anchors in the membrane and recruits effectors by exposing its effector lobe. With the use of NMR and molecular dynamics (MD) simulations, we observed that in solution, farnesylated guanosine 5′-diphosphate (GDP)-bound K-Ras4B is predominantly autoinhibited by its hypervariable region (HVR), whereas the GTP-bound state favors an activated, HVR-released state. On the anionic membrane, the catalytic domain adopts multiple orientations, including parallel (∼180°) and perpendicular (∼90°) alignments of the allosteric helices, with respect to the membrane surface direction. In the autoinhibited state, the HVR is sandwiched between the effector lobe and the membrane; in the active state, with membrane-anchored farnesyl and unrestrained HVR, the catalytic domain fluctuates reinlessly, exposing its effector-binding site. Dimerization and clustering can reduce the fluctuations. This achieves preorganized, productive conformations. Notably, we also observe HVR-autoinhibited K-Ras4B-GTP states, with GDP-bound-like orientations of the helices. Thus, we propose that the GDP/GTP exchange may not be sufficient for activation; instead, our results suggest that the GDP/GTP exchange, HVR sequestration, farnesyl insertion, and orientation/localization of the catalytic domain at the membrane conjointly determine the active or inactive state of K-Ras4B. Importantly, K-Ras4B-GTP can exist in active and inactive states; on its own, GTP binding may not compel K-Ras4B activation.—Jang, H., Banerjee, A., Chavan, T. S, Lu, S., Zhang, J., Gaponenko, V., Nussinov, R. The higher level of complexity of K-Ras4B activation at the membrane. PMID:26718888

  5. The higher level of complexity of K-Ras4B activation at the membrane.

    Science.gov (United States)

    Jang, Hyunbum; Banerjee, Avik; Chavan, Tanmay S; Lu, Shaoyong; Zhang, Jian; Gaponenko, Vadim; Nussinov, Ruth

    2016-04-01

    Is nucleotide exchange sufficient to activate K-Ras4B? To signal, oncogenic rat sarcoma (Ras) anchors in the membrane and recruits effectors by exposing its effector lobe. With the use of NMR and molecular dynamics (MD) simulations, we observed that in solution, farnesylated guanosine 5'-diphosphate (GDP)-bound K-Ras4B is predominantly autoinhibited by its hypervariable region (HVR), whereas the GTP-bound state favors an activated, HVR-released state. On the anionic membrane, the catalytic domain adopts multiple orientations, including parallel (∼180°) and perpendicular (∼90°) alignments of the allosteric helices, with respect to the membrane surface direction. In the autoinhibited state, the HVR is sandwiched between the effector lobe and the membrane; in the active state, with membrane-anchored farnesyl and unrestrained HVR, the catalytic domain fluctuates reinlessly, exposing its effector-binding site. Dimerization and clustering can reduce the fluctuations. This achieves preorganized, productive conformations. Notably, we also observe HVR-autoinhibited K-Ras4B-GTP states, with GDP-bound-like orientations of the helices. Thus, we propose that the GDP/GTP exchange may not be sufficient for activation; instead, our results suggest that the GDP/GTP exchange, HVR sequestration, farnesyl insertion, and orientation/localization of the catalytic domain at the membrane conjointly determine the active or inactive state of K-Ras4B. Importantly, K-Ras4B-GTP can exist in active and inactive states; on its own, GTP binding may not compel K-Ras4B activation.-Jang, H., Banerjee, A., Chavan, T. S, Lu, S., Zhang, J., Gaponenko, V., Nussinov, R. The higher level of complexity of K-Ras4B activation at the membrane. © FASEB.

  6. Active rehabilitation and physical therapy during extracorporeal membrane oxygenation while awaiting lung transplantation: a practical approach.

    Science.gov (United States)

    Turner, David A; Cheifetz, Ira M; Rehder, Kyle J; Williford, W Lee; Bonadonna, Desiree; Banuelos, Scott J; Peterson-Carmichael, Stacey; Lin, Shu S; Davis, R Duane; Zaas, David

    2011-12-01

    Extracorporeal membrane oxygenation as a bridge to lung transplantation has traditionally been associated with substantial morbidity and mortality. A major contributor to these complications may be weakness and overall deconditioning secondary to pretransplant critical illness and immobility. In an attempt to address this issue, we developed a collaborative program to allow for active rehabilitation and physical therapy for patients requiring life support with extracorporeal membrane oxygenation before lung transplantation. An interdisciplinary team responded to an acute need to develop a mechanism for active rehabilitation and physical therapy for patients awaiting lung transplantation while being managed with extracorporeal membrane oxygenation. We describe a series of three patients who benefited from this new approach. A quaternary care pediatric intensive care unit in a children's hospital set within an 800-bed university academic hospital with an active lung transplantation program for adolescent and adult patients. PATIENTS, INTERVENTIONS, AND MAIN RESULTS: Three patients (ages 16, 20, and 24 yrs) with end-stage respiratory failure were rehabilitated while on extracorporeal membrane oxygenation awaiting lung transplantation. These patients were involved in active rehabilitation and physical therapy and, ultimately, were ambulatory on extracorporeal membrane oxygenation before successful transplantation. Following lung transplantation, the patients were liberated from mechanical ventilation, weaned to room air, transitioned out of the intensive care unit, and ambulatory less than 1 wk posttransplant. A comprehensive, multidisciplinary system can be developed to safely allow for active rehabilitation, physical therapy, and ambulation of patients being managed with extracorporeal membrane oxygenation. Such programs may lead to a decreased threshold for the utilization of extracorporeal membrane oxygenation before transplant and have the potential to improve

  7. Cenozoic Deformation History of Asia and its Relationship to Igneous Activities

    Science.gov (United States)

    Yin, A.

    2003-04-01

    Cenozoic tectonic history of Asia has been reconstructed mostly from a forward approach based on mechanical models. Here I use the geologic data to reconstruct Asian deformation resulted from both collision of India and Arabia with Eurasia and subduction of western Pacific plates beneath Asia. Between 65 and 25 Ma, deformation of east Asia was mainly induced by the Indo-Asian collision dominated by north-south contraction in Tibet and the Himalaya. The northern contractional front of the Indo-Asian collision zone propagated northward across Tarim basin to the south Tian Shan at 25-20 Ma and to the northern Tian Shan and the Altai Shan at about 15-10 Ma. Since about 15 Ma, E-W extension began to occur throughout Asia, from Tibetan rift systems in the south to the Baikal rift system in the north. E-W extension is commonly associated with N-S contraction accommodated either by conjugate strike-slip faults or E-trending contractional structures. E-W extensional strains in Tibet and North China are kinematically linked via a large contractional step-over structure between the left-slip Qinling and Kunlun faults at the western end of the Qinling range. The Cenozoic continental-margin extension of Asia developed in two stages: (1) initially in widely distributed zone about 500-800 km wide during 65-35 Ma followed by (2) relatively localized extension associated with formation of back-arc basins floored by oceanic crust during 35-15 Ma. Since ˜15 Ma, the eastern margin of Asia became contractional as expressed by collapse of back-arc basins. This event is coeval with widespread east-west extension in the interior of mainland Asia. The Oligo-Miocene Arabia-Eurasia collision may have created a 3000-km long and 700-km wide zone of subparallel and relatively evenly spaced right-slip faults, stretching from the Main Recent Fault in NW Iran in the south to the Altai fault of western Mongolia in the north. The collision may have also assisted the initiation of the

  8. Compartmentalized system with membrane-bound glycerol kinase. Activity and product distribution versus asymmetrical substrate supply.

    Science.gov (United States)

    Girard, A; Merchie, B; Maïsterrena, B

    1991-03-15

    An artificial-membrane-bound glycerokinase chosen as a membrane-bound two-substrate-enzyme model has been used to separate two unequal compartments of a specially designed diffusion cell. An interesting feature is the asymmetry of compartments and the existence of a diffusion layer adjacent to only one face of the enzymic membrane. In such a situation the apparent enzyme activity and the product distribution in the system have been studied versus all the possibilities of combination of ATP and glycerol supply. Our approach has lead us to differentiate two different roles played by a diffusion layer adjacent to a permeable enzymic membrane. Depending on the spatial origin of the enzymic substrates (i.e. from which compartment they derive), the diffusion layer can play either the role of a passive additional resistance to that of the membrane or the role of a third compartment in which the reaction product can partially accumulate before splitting on both parts of the membrane. Our results mainly demonstrate that a membrane-bound enzyme activity and the resulting product distribution occurring in a compartmentalized system may be regulated by the cumulative effect due to the asymmetry in volumes of the compartments, the presence of a diffusion layer and the different possibilities of substrate supply. With the topography studied, which is close to that reported for many 'in vivo' situations, the product may be diffused lead to vectorial metabolism processes.

  9. Activation of Endothelial Nitric Oxide (eNOS Occurs through Different Membrane Domains in Endothelial Cells.

    Directory of Open Access Journals (Sweden)

    Jason Tran

    Full Text Available Endothelial cells respond to a large range of stimuli including circulating lipoproteins, growth factors and changes in haemodynamic mechanical forces to regulate the activity of endothelial nitric oxide synthase (eNOS and maintain blood pressure. While many signalling pathways have been mapped, the identities of membrane domains through which these signals are transmitted are less well characterized. Here, we manipulated bovine aortic endothelial cells (BAEC with cholesterol and the oxysterol 7-ketocholesterol (7KC. Using a range of microscopy techniques including confocal, 2-photon, super-resolution and electron microscopy, we found that sterol enrichment had differential effects on eNOS and caveolin-1 (Cav1 colocalisation, membrane order of the plasma membrane, caveolae numbers and Cav1 clustering. We found a correlation between cholesterol-induced condensation of the plasma membrane and enhanced high density lipoprotein (HDL-induced eNOS activity and phosphorylation suggesting that cholesterol domains, but not individual caveolae, mediate HDL stimulation of eNOS. Vascular endothelial growth factor (VEGF-induced and shear stress-induced eNOS activity was relatively independent of membrane order and may be predominantly controlled by the number of caveolae on the cell surface. Taken together, our data suggest that signals that activate and phosphorylate eNOS are transmitted through distinct membrane domains in endothelial cells.

  10. Automatic corpus callosum segmentation using a deformable active Fourier contour model.

    Science.gov (United States)

    Vachet, Clement; Yvernault, Benjamin; Bhatt, Kshamta; Smith, Rachel G; Gerig, Guido; Hazlett, Heather Cody; Styner, Martin

    2012-03-23

    The corpus callosum (CC) is a structure of interest in many neuroimaging studies of neuro-developmental pathology such as autism. It plays an integral role in relaying sensory, motor and cognitive information from homologous regions in both hemispheres. We have developed a framework that allows automatic segmentation of the corpus callosum and its lobar subdivisions. Our approach employs constrained elastic deformation of flexible Fourier contour model, and is an extension of Szekely's 2D Fourier descriptor based Active Shape Model. The shape and appearance model, derived from a large mixed population of 150+ subjects, is described with complex Fourier descriptors in a principal component shape space. Using MNI space aligned T1w MRI data, the CC segmentation is initialized on the mid-sagittal plane using the tissue segmentation. A multi-step optimization strategy, with two constrained steps and a final unconstrained step, is then applied. If needed, interactive segmentation can be performed via contour repulsion points. Lobar connectivity based parcellation of the corpus callosum can finally be computed via the use of a probabilistic CC subdivision model. Our analysis framework has been integrated in an open-source, end-to-end application called CCSeg both with a command line and Qt-based graphical user interface (available on NITRC). A study has been performed to quantify the reliability of the semi-automatic segmentation on a small pediatric dataset. Using 5 subjects randomly segmented 3 times by two experts, the intra-class correlation coefficient showed a superb reliability (0.99). CCSeg is currently applied to a large longitudinal pediatric study of brain development in autism.

  11. Automatic corpus callosum segmentation using a deformable active Fourier contour model

    Science.gov (United States)

    Vachet, Clement; Yvernault, Benjamin; Bhatt, Kshamta; Smith, Rachel G.; Gerig, Guido; Cody Hazlett, Heather; Styner, Martin

    2012-03-01

    The corpus callosum (CC) is a structure of interest in many neuroimaging studies of neuro-developmental pathology such as autism. It plays an integral role in relaying sensory, motor and cognitive information from homologous regions in both hemispheres. We have developed a framework that allows automatic segmentation of the corpus callosum and its lobar subdivisions. Our approach employs constrained elastic deformation of flexible Fourier contour model, and is an extension of Szekely's 2D Fourier descriptor based Active Shape Model. The shape and appearance model, derived from a large mixed population of 150+ subjects, is described with complex Fourier descriptors in a principal component shape space. Using MNI space aligned T1w MRI data, the CC segmentation is initialized on the mid-sagittal plane using the tissue segmentation. A multi-step optimization strategy, with two constrained steps and a final unconstrained step, is then applied. If needed, interactive segmentation can be performed via contour repulsion points. Lobar connectivity based parcellation of the corpus callosum can finally be computed via the use of a probabilistic CC subdivision model. Our analysis framework has been integrated in an open-source, end-to-end application called CCSeg both with a command line and Qt-based graphical user interface (available on NITRC). A study has been performed to quantify the reliability of the semi-automatic segmentation on a small pediatric dataset. Using 5 subjects randomly segmented 3 times by two experts, the intra-class correlation coefficient showed a superb reliability (0.99). CCSeg is currently applied to a large longitudinal pediatric study of brain development in autism.

  12. MEMS Deformable Mirrors for Adaptive Optics in Astronomical Imaging

    Science.gov (United States)

    Cornelissen, S.; Bierden, P. A.; Bifano, T.

    We report on the development of micro-electromechanical (MEMS) deformable mirrors designed for ground and space-based astronomical instruments intended for imaging extra-solar planets. Three different deformable mirror designs, a 1024 element continuous membrane (32x32), a 4096 element continuous membrane (64x64), and a 331 hexagonal segmented tip-tilt-piston are being produced for the Planet Imaging Concept Testbed Using a Rocket Experiment (PICTURE) program, the Gemini Planet Imaging Instrument, and the visible nulling coronograph developed at JPL for NASA's TPF mission, respectively. The design of these polysilicon, surface-micromachined MEMS deformable mirrors builds on technology that was pioneered at Boston University and has been used extensively to correct for ocular aberrations in retinal imaging systems and for compensation of atmospheric turbulence in free-space laser communication. These light-weight, low power deformable mirrors will have an active aperture of up to 25.2mm consisting of thin silicon membrane mirror supported by an array of 1024 to 4096 electrostatic actuators exhibiting no hysteresis and sub-nanometer repeatability. The continuous membrane deformable mirrors, coated with a highly reflective metal film, will be capable of up to 4μm of stroke, have a surface finish of travel. New design features and fabrication processes are combined with a proven device architecture to achieve the desired performance and high reliability. Presented in this paper are device characteristic and performance results of these devices.

  13. Receptor kinase-mediated control of primary active proton pumping at the plasma membrane

    DEFF Research Database (Denmark)

    Fuglsang, Anja Thoe; Kristensen, Astrid; Cuin, Tracey A.

    2014-01-01

    Acidification of the cell wall space outside the plasma membrane is required for plant growth and is the result of proton extrusion by the plasma membrane-localized H+-ATPases. Here we show that the major plasma membrane proton pumps in Arabidopsis, AHA1 and AHA2, interact directly in vitro...... heterologous expression system, the introduction of a negative charge at this position caused pump activation. Application of PSY1 to plant seedlings induced rapid in planta phosphorylation at Thr-881, concomitant with an instantaneous increase in proton efflux from roots. The direct interaction between AHA2...

  14. Nonlinear Dielectric Spectroscopy as an Indirect Probe of Metabolic Activity in Thylakoid Membrane

    Directory of Open Access Journals (Sweden)

    John H. Miller

    2011-01-01

    Full Text Available Nonlinear dielectric spectroscopy (NDS is a non-invasive probe of cellular metabolic activity with potential application in the development of whole-cell biosensors. However, the mechanism of NDS interaction with metabolic membrane proteins is poorly understood, partly due to the inherent complexity of single cell organisms. Here we use the light-activated electron transport chain of spinach thylakoid membrane as a model system to study how NDS interacts with metabolic activity. We find protein modification, as opposed to membrane pump activity, to be the dominant source of NDS signal change in this system. Potential mechanisms for such protein modifications include reactive oxygen species generation and light-activated phosphorylation.

  15. Caffeine inhibits erythrocyte membrane derangement by antioxidant activity and by blocking caspase 3 activation.

    Science.gov (United States)

    Tellone, Ester; Ficarra, Silvana; Russo, Annamaria; Bellocco, Ersilia; Barreca, Davide; Laganà, Giuseppina; Leuzzi, Ugo; Pirolli, Davide; De Rosa, Maria Cristina; Giardina, Bruno; Galtieri, Antonio

    2012-02-01

    The aim of this research was to investigate the effect of caffeine on band 3 (the anion exchanger protein), haemoglobin function, caspase 3 activation and glucose-6-phosphate metabolism during the oxygenation-deoxygenation cycle in human red blood cells. A particular attention has been given to the antioxidant activity by using in vitro antioxidant models. Caffeine crosses the erythrocyte membrane and interacts with the two extreme conformational states of haemoglobin (the T and the R-state within the framework of the simple two states allosteric model) with different binding affinities. By promoting the high affinity state (R-state), the caffeine-haemoglobin interaction does enhance the pentose phosphate pathway. This is of benefit for red blood cells since it leads to an increase of NADPH availability. Moreover, caffeine effect on band 3, mediated by haemoglobin, results in an extreme increase of the anion exchange, particularly in oxygenated erythrocytes. This enhances the transport of the endogenously produced CO(2) thereby avoiding the production of dangerous secondary radicals (carbonate and nitrogen dioxide) which are harmful to the cellular membrane. Furthermore caffeine destabilizes the haeme-protein interactions within the haemoglobin molecule and triggers the production of superoxide and met-haemoglobin. However this damaging effect is almost balanced by the surprising scavenger action of the alkaloid with respect to the hydroxyl radical. These experimental findings are supported by in silico docking and molecular dynamics studies and by what we may call the "caspase silence"; in fact, there is no evidence of any caspase 3 activity enhancement; this is likely due to the promotion of positive metabolic conditions which result in an increase of the cellular reducing power. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

  16. Improved antibacterial activity of nanofiltration polysulfone membranes modified with silver nanoparticles.

    Science.gov (United States)

    Andrade, Patricia Fernanda; de Faria, Andreia Fonseca; Oliveira, Silvana Ruella; Arruda, Marco Aurélio Zezzi; Gonçalves, Maria do Carmo

    2015-09-15

    Polysulfone membranes (PSf) containing silver nanoparticles were prepared by the wet phase-inversion process. Silver nanoparticles (AgNP) were dispersed into the polymer matrix using two different methodologies. In the first one, the AgNP were synthesized and further dispersed into the polymer solution (ex situ process). In the second method, the formation of the AgNP was performed in situ. The AgNP crystalline structure in the PSf membranes was confirmed by X-ray diffraction. Field emission scanning electron microscopy images showed that the addition of AgNP in PSf membranes caused no significant changes to the finger-like morphology. When the ex situ methodology was applied, 45 nm average size AgNP were uniformly distributed in the internal pores of the membranes. However, when the AgNP were formed through the in situ process, the AgNP were uniformly and preferentially distributed on the top and bottom surfaces of the membrane. In the last case, the AgNP showed cubic morphology when present in the bottom and top surfaces, however, when inside the membrane their morphology was spherical. The cubic-like nanoparticles displayed a 38 nm average edge length. The silver ion released from the membrane during water filtration was measured using inductively coupled plasma mass spectrometry, which showed a silver leaching of approximately 2 μg L(-1). The nanocomposite membranes prepared by the in situ method exhibited a better antibacterial activity, in comparison to those prepared by ex situ, and also a decrease in 90% Escherichia coli adhered cells compared to the pristine PSf membranes. In conclusion, the in situ procedure can be considered a feasible, simple, and reproducible methodology to prepare anti-biofouling polysulfone membranes containing AgNP. Copyright © 2015 Elsevier Ltd. All rights reserved.

  17. Tuning Liposome Membrane Permeability by Competitive Peptide Dimerization and Partitioning-Folding Interactions Regulated by Proteolytic Activity

    Science.gov (United States)

    Lim, Seng Koon; Sandén, Camilla; Selegård, Robert; Liedberg, Bo; Aili, Daniel

    2016-02-01

    Membrane active peptides are of large interest for development of drug delivery vehicles and therapeutics for treatment of multiple drug resistant infections. Lack of specificity can be detrimental and finding routes to tune specificity and activity of membrane active peptides is vital for improving their therapeutic efficacy and minimize harmful side effects. We describe a de novo designed membrane active peptide that partition into lipid membranes only when specifically and covalently anchored to the membrane, resulting in pore-formation. Dimerization with a complementary peptide efficiently inhibits formation of pores. The effect can be regulated by proteolytic digestion of the inhibitory peptide by the matrix metalloproteinase MMP-7, an enzyme upregulated in many malignant tumors. This system thus provides a precise and specific route for tuning the permeability of lipid membranes and a novel strategy for development of recognition based membrane active peptides and indirect enzymatically controlled release of liposomal cargo.

  18. Biological Activity of Blackcurrant Extracts (Ribes nigrum L. in Relation to Erythrocyte Membranes

    Directory of Open Access Journals (Sweden)

    Dorota Bonarska-Kujawa

    2014-01-01

    Full Text Available Compounds contained in fruits and leaves of blackcurrant (Ribes nigrum L. are known as agents acting preventively and therapeutically on the organism. The HPLC analysis showed they are rich in polyphenol anthocyanins in fruits and flavonoids in leaves, that have antioxidant activity and are beneficial for health. The aim of the research was to determine the effect of blackcurrant fruit and leaf extracts on the physical properties of the erythrocyte membranes and assess their antioxidant properties. The effect of the extracts on osmotic resistance, shape of erythrocytes and hemolytic and antioxidant activity of the extracts were examined with spectrophotometric methods. The FTIR investigation showed that extracts modify the erythrocyte membrane and protect it against free radicals induced by UV radiation. The results show that the extracts do not induce hemolysis and even protect erythrocytes against the harmful action of UVC radiation, while slightly strengthening the membrane and inducing echinocytes. The compounds contained in the extracts do not penetrate into the hydrophobic region, but bind to the membrane surface inducing small changes in the packing arrangement of the polar head groups of membrane lipids. The extracts have a high antioxidant activity. Their presence on the surface of the erythrocyte membrane entails protection against free radicals.

  19. Radionuclide assay of membrane Na+, K+-ATPase activity of peserved red blood cells

    International Nuclear Information System (INIS)

    Trusov, V.V.; Zelenin, A.A.; Marizin, S.A.

    1986-01-01

    The radionuclide tests were used to investigate the influence of varying blood preservatives on erythrocylic membrane Na + , K + -ATPase activity in samples of whole blood and packed red blood cells from normal donors prepared by standard methods. The tests were performed before and after seven days of preservation under standard conditions. It was found that blood preservations lowered membrane Na + , K + -ATPase activity: its minimum reduction was recorded with citroglucopnosphate, while glugicir induced a significant drop in Na + , K + -ATPase activity of preserved red blood cells regardless of the type of the blood transfusion solution. The assay of membrane Na + , K + -ATPase activity of preserved red blood cells with the use of 86 Rb could be recommended as an evaluation test for preserved blood and its components

  20. Sphingomyelinase D activity in model membranes: structural effects of in situ generation of ceramide-1-phosphate.

    Directory of Open Access Journals (Sweden)

    Roberto P Stock

    Full Text Available The toxicity of Loxosceles spider venom has been attributed to a rare enzyme, sphingomyelinase D, which transforms sphingomyelin to ceramide-1-phosphate. The bases of its inflammatory and dermonecrotic activity, however, remain unclear. In this work the effects of ceramide-1-phosphate on model membranes were studied both by in situ generation of this lipid using a recombinant sphingomyelinase D from the spider Loxosceles laeta and by pre-mixing it with sphingomyelin and cholesterol. The systems of choice were large unilamellar vesicles for bulk studies (enzyme kinetics, fluorescence spectroscopy and dynamic light scattering and giant unilamellar vesicles for fluorescence microscopy examination using a variety of fluorescent probes. The influence of membrane lateral structure on the kinetics of enzyme activity and the consequences of enzyme activity on the structure of target membranes containing sphingomyelin were examined. The findings indicate that: 1 ceramide-1-phosphate (particularly lauroyl ceramide-1-phosphate can be incorporated into sphingomyelin bilayers in a concentration-dependent manner and generates coexistence of liquid disordered/solid ordered domains, 2 the activity of sphingomyelinase D is clearly influenced by the supramolecular organization of its substrate in membranes and, 3 in situ ceramide-1-phosphate generation by enzymatic activity profoundly alters the lateral structure and morphology of the target membranes.

  1. Sphingomyelinase D activity in model membranes: structural effects of in situ generation of ceramide-1-phosphate.

    Science.gov (United States)

    Stock, Roberto P; Brewer, Jonathan; Wagner, Kerstin; Ramos-Cerrillo, Blanca; Duelund, Lars; Jernshøj, Kit Drescher; Olsen, Lars Folke; Bagatolli, Luis A

    2012-01-01

    The toxicity of Loxosceles spider venom has been attributed to a rare enzyme, sphingomyelinase D, which transforms sphingomyelin to ceramide-1-phosphate. The bases of its inflammatory and dermonecrotic activity, however, remain unclear. In this work the effects of ceramide-1-phosphate on model membranes were studied both by in situ generation of this lipid using a recombinant sphingomyelinase D from the spider Loxosceles laeta and by pre-mixing it with sphingomyelin and cholesterol. The systems of choice were large unilamellar vesicles for bulk studies (enzyme kinetics, fluorescence spectroscopy and dynamic light scattering) and giant unilamellar vesicles for fluorescence microscopy examination using a variety of fluorescent probes. The influence of membrane lateral structure on the kinetics of enzyme activity and the consequences of enzyme activity on the structure of target membranes containing sphingomyelin were examined. The findings indicate that: 1) ceramide-1-phosphate (particularly lauroyl ceramide-1-phosphate) can be incorporated into sphingomyelin bilayers in a concentration-dependent manner and generates coexistence of liquid disordered/solid ordered domains, 2) the activity of sphingomyelinase D is clearly influenced by the supramolecular organization of its substrate in membranes and, 3) in situ ceramide-1-phosphate generation by enzymatic activity profoundly alters the lateral structure and morphology of the target membranes.

  2. Influence of Active Layer on Separation Potentials of Nanofiltration Membranes for Inorganic Ions.

    Science.gov (United States)

    Wadekar, Shardul S; Vidic, Radisav D

    2017-05-16

    Active layers of two fully aromatic and two semi-aromatic nanofiltration membranes were studied along with surface charge at different electrolyte composition and effective pore size to elucidate their influence on separation mechanisms for inorganic ions by steric, charge, and dielectric exclusion. The membrane potential method used for pore size measurement is underlined as the most appropriate measurement technique for this application owing to its dependence on the diffusional potentials of inorganic ions. Crossflow rejection experiments with dilute feed composition indicate that both fully aromatic membranes achieved similar rejection despite the differences in surface charge, which suggests that rejection by these membranes is exclusively dependent on size exclusion and the contribution of charge exclusion is weak. Rejection experiments with higher ionic strength and different composition of the feed solution confirmed this hypothesis. On the other hand, increase in the ionic strength of feed solution when the charge exclusion effects are negligible due to charge screening strongly influenced ion rejection by semi-aromatic membranes. The experimental results confirmed that charge exclusion contributes significantly to the performance of semi-aromatic membranes in addition to size exclusion. The contribution of dielectric exclusion to overall ion rejection would be more significant for fully aromatic membranes.

  3. Preparation of amino-functionalized regenerated cellulose membranes with high catalytic activity.

    Science.gov (United States)

    Wang, Wei; Bai, Qian; Liang, Tao; Bai, Huiyu; Liu, Xiaoya

    2017-09-01

    The modification of regenerated cellulose (RC) membranes was carried out by using silane coupling agents presenting primary and secondary amino-groups. The grafting of the amino groups onto the modified cellulose molecule was confirmed by X-ray photoelectron spectroscopies and 13 C nuclear magnetic resonance spectroscopic analyses. The crystallinity of the cellulose membranes (CM) decreased after chemical modification as indicated by the X-ray diffraction results. Moreover, a denser structure was observed at the surface and cross section of the modified membranes by SEM images. The contact angle measurements showed that the silane coupling treatment enhanced the hydrophobicity of the obtained materials. Then the catalytic properties of two types of modified membranes were studied in a batch process by evaluating their catalytic performance in a Knoevenagel condensation. The results indicated that the cellulose membrane grafted with many secondary amines exhibited a better catalytic activity compared to the one grafted only by primary amines. In addition, the compact structure of the modified membranes permitted their application in a pervaporation catalytic membrane reactor. Therefore, functional CM that prepared in this paper represented a promising material in the field of industrial catalysis. Copyright © 2017 Elsevier B.V. All rights reserved.

  4. Effect of various concentration of sulfuric acid for Nafion membrane activation on the performance of fuel cell

    Science.gov (United States)

    Pujiastuti, Sri; Onggo, Holia

    2016-02-01

    This work proposes an activation treatment to Nafion 117 membrane with sulfuric acid in various concentrations. The main goal of this study is to increase the Nafion 117 membrane performance, which is determined by proton number in the membrane and membrane performance in Polymer Electrolyte Membrane Fuel Cell (PEMFC). This work was developed using sulfuric acids in four different concentrations: 1, 2, 3, and 4 M. The surface morphology and functional groups of activated membranes were studied using Scanning Electron Microscope and Fourier Transform Infrared, respectively. The proton number absorbed in membranes was observed by gravimetric measurements. The performances of activated membranes in PEMFC were studied by single cell measurements with H2/O2 operation. The experimental results showed that activation of Nafion membrane did not change its surface morphology and functional groups. The proton number increased when the concentration of sulfuric acid is increased from 1 to 3 M and from 1 to 4 M. On the other hand, there is no significant increase when the concentration of sulfuric acid was increased from 1 to 2 M. Similar trends were observed when testing activated membrane performance in PEMFC, especially for current density at 0.6 V and maximum power. It is assumed that there is a correlation between the increase of sulfuric acid concentration in activation process with the increase of proton number in the membrane that are available for facilitating of transfer protons from the anode to the cathode.

  5. Membranolytic Activity of Bile Salts: Influence of Biological Membrane Properties and Composition

    Directory of Open Access Journals (Sweden)

    Alfred Blume

    2007-10-01

    Full Text Available The two main steps of the membranolytic activity of detergents: 1 the partitioning of detergent molecules in the membrane and 2 the solubilisation of the membrane are systematically investigated. The interactions of two bile salt molecules, sodium cholate (NaC and sodium deoxycholate (NaDC with biological phospholipid model membranes are considered. The membranolytic activity is analysed as a function of the hydrophobicity of the bile salt, ionic strength, temperature, membrane phase properties, membrane surface charge and composition of the acyl chains of the lipids. The results are derived from calorimetric measurements (ITC, isothermal titration calorimetry. A thermodynamic model is described, taking into consideration electrostatic interactions, which is used for the calculation of the partition coefficient as well as to derive the complete thermodynamic parameters describing the interaction of detergents with biological membranes (change in enthalpy, change in free energy, change in entropy etc. The solubilisation properties are described in a so-called vesicle-to-micelle phase transition diagram. The obtained results are supplemented and confirmed by data obtained from other biophysical techniques (DSC differential scanning calorimetry, DLS dynamic light scattering, SANS small angle neutron scattering.

  6. Protein kinase and phosphatase activities of thylakoid membranes

    International Nuclear Information System (INIS)

    Michel, H.; Shaw, E.K.; Bennett, J.

    1987-01-01

    Dephosphorylation of the 25 and 27 kDa light-harvesting Chl a/b proteins (LHCII) of the thylakoid membranes is catalyzed by a phosphatase which differs from previously reported thylakoid-bound phosphatases in having an alkaline pH optimum (9.0) and a requirement for Mg 2+ ions. Dephosphorylation of the 8.3 kDa psb H gene product requires a Mg 2+ ion concentration more than 200 fold higher than that for dephosphorylation of LHC II. The 8.3 kDa and 27 kDa proteins appear to be phosphorylated by two distinct kinases, which differ in substrate specificity and sensitivity to inhibitors. The plastoquinone antagonist 2,5-dibromo-3-methyl-6-isopropyl-benzoquinone (DBMIB) inhibits phosphorylation of the 27 kDa LHC II much more readily than phosphorylation of the 8.3 kDa protein. A similar pattern of inhibition is seen for two synthetic oligopeptides (MRKSATTKKAVC and ATQTLESSSRC) which are analogs of the phosphorylation sites of the two proteins. Possible modes of action of DBMIB are discussed. 45 refs., 7 figs., 3 tabs

  7. Membrane-associated proteolytic activity in Escherichia coli that is stimulated by ATP

    International Nuclear Information System (INIS)

    Klemes, Y.; Voellmy, R.W.; Goldberg, A.L.

    1986-01-01

    The degradation of proteins in bacteria requires metabolism energy. One important enzyme in this process is protease La, a soluble ATP-dependent protease encoded by the lon gene. However, lon mutants that lack a functional protease La still show some ATP-dependent protein breakdown. The authors have reported an ATP-stimulated endoproteolytic activity associated with the inner membrane of E. coli. This ATP-stimulated activity is found in normal levels in membranes derived from lon mutants, including strains carrying insertions in the lon gene. The membrane-bound activity hydrolyzes 14 C-methylglobin at a linear rate for up to 3 hours. These fractions also contain appreciable proteolytic activity that is not affected by ATP. The stimulation by ATP requires the presence of Mg 2+ . Nonhydrolyzable ATP analogs (e.g. AMPPNP or ATP-γ-S) and ADP do not enhance proteolysis. Unlike protease La, the membrane-associated enzyme does not degrade the fluorometric substrate, Glt-Ala-Ala-Phe-MNA, in an ATP-stimulated fashion, and its level is not influenced by high temperature of by the gene which regulates the heat-shock response. The enzyme is inhibited by dichloroisocoumarin and certain peptide chloromethyl ketones. They conclude that E. coli contain at least two ATP-dependent proteases with distinct specificities: one is soluble and the other is membrane-associated

  8. Spontaneous membrane formation and self-encapsulation of active rods in an inhomogeneous motility field

    Science.gov (United States)

    Grauer, Jens; Löwen, Hartmut; Janssen, Liesbeth M. C.

    2018-02-01

    We study the collective dynamics of self-propelled rods in an inhomogeneous motility field. At the interface between two regions of constant but different motility, a smectic rod layer is spontaneously created through aligning interactions between the active rods, reminiscent of an artificial, semipermeable membrane. This "active membrane" engulfes rods which are locally trapped in low-motility regions and thereby further enhances the trapping efficiency by self-organization, an effect which we call "self-encapsulation." Our results are gained by computer simulations of self-propelled rod models confined on a two-dimensional planar or spherical surface with a stepwise constant motility field, but the phenomenon should be observable in any geometry with sufficiently large spatial inhomogeneity. We also discuss possibilities to verify our predictions of active-membrane formation in experiments of self-propelled colloidal rods and vibrated granular matter.

  9. Modulation of the activities of membrane enzymes by cereal grain resorcinolic lipids.

    Science.gov (United States)

    Kozubek, A; Nietubyc, M; Sikorski, A F

    1992-01-01

    Resorcinolic lipids, amphiphilic compounds from cereal grains show strong effects upon the activity of membrane enzymes. The concentrations for 50% inhibition of erythrocyte membrane acetylcholinesterase were in the range of 18-90 microM and were dependent on the length of the aliphatic side chain of the homologue and on the modification of hydroxyl groups in the benzene ring. Sulfonation of OH groups resulted in a drastic decrease of the inhibitory potency. The effect of resorcinolic lipids on the activity of Ca2+(calmodulin)-ATPase was the opposite. Up to concentrations of 50 microM alk(en)ylresorcinols stimulated the activity of this enzyme and only slight inhibition (approx. 30%) was observed above 100 microM. The results suggest that the effect of resorcinolic lipids might depend on their ability to alter the bilayer properties. Most probably these compounds decrease the mobility of membrane phospholipid molecules.

  10. Sphingomyelinase D activity in model membranes: structural effects of in situ generation of ceramide-1-phosphate

    DEFF Research Database (Denmark)

    Stock, Roberto; Brewer, Jonathan R.; Wagner, Kerstin

    2012-01-01

    The toxicity of Loxosceles spider venom has been attributed to a rare enzyme, sphingomyelinase D, which transforms sphingomyelin to ceramide-1-phosphate. The bases of its inflammatory and dermonecrotic activity, however, remain unclear. In this work the effects of ceramide-1-phosphate on model...... membranes were studied both by in situ generation of this lipid using a recombinant sphingomyelinase D from the spider Loxosceles laeta and by pre-mixing it with sphingomyelin and cholesterol. The systems of choice were large unilamellar vesicles for bulk studies (enzyme kinetics, fluorescence spectroscopy...... and dynamic light scattering) and giant unilamellar vesicles for fluorescence microscopy examination using a variety of fluorescent probes. The influence of membrane lateral structure on the kinetics of enzyme activity and the consequences of enzyme activity on the structure of target membranes containing...

  11. Active folding of fluvial terraces across a `blind' Himalayan deformation front in the Kashmir Himalaya, northwest India.

    Science.gov (United States)

    Gavillot, Y. G.; Meigs, A.; Rittenour, T. M.; Malik, M. O. A.

    2016-12-01

    In Kashmir, the Himalayan Frontal thrust (HFT) is blind, characterized by a broad fold, the Suruin-Mastargh anticline, and displays no emergent faults cutting either limb. A lack of knowledge of the rate of shortening and structural framework of the Suruin-Mastargh anticline hampers quantifying the earthquake potential for the deformation front. Our study utilized the geomorphic expression of dated deformed terraces on the Ujh River in Kashmir. Six terraces are recognized, and four yield multiple optically stimulated luminescence (OSL) and depth profiles terrigenous cosmogenic nuclides (TCN) ages between 53 ka and 0.4 ka. Vector fold restoration of long terrace profiles indicates a deformation pattern characterized by regional uplift across the anticlinal axis and back-limb, and by fold limb rotation on the forelimb. Differential uplift across the fold trace suggests localized deformation. Dip data and stratigraphic thicknesses suggest that a duplex structure is emplaced at depth along the basal décollement, folding the overlying roof thrust and Siwalik-Murree strata into a detachment-like fold. Localized faulting at the fold axis explains the asymmetrical fold geometry. Folding of the oldest dated terrace, suggests rock uplift rates across the Suruin-Mastargh anticline range between 1.8-2.5 mm/yr. Assuming a 25° dipping ramp for the blind structure on the basis of dip data constraints, the shortening rates across the Suruin-Mastargh anticline range between 3.8-5.4 mm/yr since 53 ka. Geodetic data indicate that an 11-12 mm/yr arc-normal shortening rate characterizes the interseismic strain accumulation across the plate boundary due to India-Tibet convergence. These data combined with rates of other active internal faults in the Kashmir Himalaya indicate that the Riasi fault accounts for the remainder 60% of the convergence not taken up by the Suruin-Mastargh anticline. We attribute a non-emergent thrust at the deformation front to reflect deformation controlled

  12. Membrane-active macromolecules resensitize NDM-1 gram-negative clinical isolates to tetracycline antibiotics.

    Directory of Open Access Journals (Sweden)

    Divakara S S M Uppu

    Full Text Available Gram-negative 'superbugs' such as New Delhi metallo-beta-lactamase-1 (blaNDM-1 producing pathogens have become world's major public health threats. Development of molecular strategies that can rehabilitate the 'old antibiotics' and halt the antibiotic resistance is a promising approach to target them. We report membrane-active macromolecules (MAMs that restore the antibacterial efficacy (enhancement by >80-1250 fold of tetracycline antibiotics towards blaNDM-1 Klebsiella pneumonia and blaNDM-1 Escherichia coli clinical isolates. Organismic studies showed that bacteria had an increased and faster uptake of tetracycline in the presence of MAMs which is attributed to the mechanism of re-sensitization. Moreover, bacteria did not develop resistance to MAMs and MAMs stalled the development of bacterial resistance to tetracycline. MAMs displayed membrane-active properties such as dissipation of membrane potential and membrane-permeabilization that enabled higher uptake of tetracycline in bacteria. In-vivo toxicity studies displayed good safety profiles and preliminary in-vivo antibacterial efficacy studies showed that mice treated with MAMs in combination with antibiotics had significantly decreased bacterial burden compared to the untreated mice. This report of re-instating the efficacy of the antibiotics towards blaNDM-1 pathogens using membrane-active molecules advocates their potential for synergistic co-delivery of antibiotics to combat Gram-negative superbugs.

  13. Correction: Membrane-active macromolecules resensitize NDM-1 gram-negative clinical isolates to tetracycline antibiotics.

    Directory of Open Access Journals (Sweden)

    Divakara S S M Uppu

    Full Text Available Gram-negative 'superbugs' such as New Delhi metallo-beta-lactamase-1 (blaNDM-1 producing pathogens have become world's major public health threats. Development of molecular strategies that can rehabilitate the 'old antibiotics' and halt the antibiotic resistance is a promising approach to target them. We report membrane-active macromolecules (MAMsthat restore the antibacterial efficacy (enhancement by >80-1250 fold of tetracycline antibiotics towards blaNDM-1 Klebsiella pneumonia and blaNDM-1 Escherichia coli clinical isolates.Organismic studies showed that bacteria had an increased and faster uptake of tetracyclinein the presence of MAMs which is attributed to the mechanism of re-sensitization. Moreover,bacteria did not develop resistance to MAMs and MAMs stalled the development of bacterial resistance to tetracycline. MAMs displayed membrane-active properties such as dissipation of membrane potential and membrane-permeabilization that enabled higher uptake of tetracycline in bacteria. In-vivo toxicity studies displayed good safety profiles and preliminary in-vivo antibacterial efficacy studies showed that mice treated with MAMs in combination with antibiotics had significantly decreased bacterial burden compared to the untreated mice. This report of re-instating the efficacy of the antibiotics towards blaNDM-1 pathogens using membrane-active molecules advocates their potential for synergistic co-delivery of antibiotics to combat Gram-negative superbugs.

  14. Antibiofilm activity of Bacillus pumilus SW9 against initial biofouling on microfiltration membranes.

    Science.gov (United States)

    Zhang, Ying; Yu, Xin; Gong, Song; Ye, Chengsong; Fan, Zihong; Lin, Huirong

    2014-02-01

    Membrane biofouling, resulting from biofilm formation on the membrane, has become the main obstacle hindering wider application of membrane technology. Initial biofouling proves to be crucial which involves early stages of microbial adhesion and biofilm formation. Biological control of microbial attachment seems to be a promising strategy due to its high efficiency and eco-friendliness. The present study investigated the effects of a bacterium Bacillus pumilus SW9 on controlling the initial fouling formed by four target bacterial strains which were pioneer species responsible for biofouling in membrane bioreactors, using microfiltration membranes as the abiotic surfaces. The results suggested that strain SW9 exhibited excellent antibiofilm activity by decreasing the attached biomass of target strains. The production of extracellular polysaccharides and proteins by four target strains was also reduced. A distinct improvement of permeate flux in dead-end filtration systems was achieved when introducing strain SW9 to microfiltration experiments. Scanning electron microscopy and confocal laser scanning microscopy were performed to further ascertain significant changes of the biofouling layers. A link between biofilm inhibition and initial biofouling mitigation was thus provided, suggesting an alternatively potential way to control membrane biofouling through bacterial interactions.

  15. [Biomechanic and biological activity assessment of concavity-convex amniotic membrane].

    Science.gov (United States)

    Shao, Yi; Yu, Yao; Tan, Gang; Peng, Juan; Zhou, Qiong; Pei, Chonggang; Dong, Wenjia; Gao, Guiping

    2012-12-01

    This paper conducted research on biomechanical characteristics and biological activity of concavity-convex amniotic membrane (CCAM) and discussed its superiority as ocular surface repair material. Folding and compression with vacuum of fresh amniotic membrane were used to prepare CCAM. After cutting the striga of CCAM, sixteen CCAM tissue section were chosen at random to test their tensile strength using electronic universal testing machine. The bilayer amniotic membrane (BAM), the double-deck amniotic membrane (DAM) and the monolayer amniotic membrane (MAM) were as controls. The test parameters included yield strength, tensile strength, elongation at break, elastic modulus and so on. The cytokines of fresh amniotic membrane (FAM), MAM and CCAM were analyzed by radioimmunoassay method. The CCAM was obviously thicker than MAM and DAM. After 15 min in PBS, the CCAM tissue can recover the normal shape. The tensile strength and the elongation at break of CCAM were higher than those of the MAM and the DAM (P biomechanical properties than the MAM and the DAM, showing the superiority as ocular surface repair material.

  16. Electrolyte Composition of Mink (Mustela vison Erythrocytes and Active Cation Transporters of the Cell Membrane

    Directory of Open Access Journals (Sweden)

    Clausen TN

    2001-06-01

    Full Text Available Red blood cells from mink (Mustela vison were characterized with respect to their electrolyte content and their cell membranes with respect to enzymatic activity for cation transport. The intra- and extracellular concentrations of Na+, K+, Cl-, Ca2+ and Mg2+ were determined in erythrocytes and plasma, respectively. Plasma and red cell water content was determined, and molal electrolyte concentrations were calculated. Red cells from male adult mink appeared to be of the low-K+, high-Na+ type as seen in other carnivorous species. The intracellular K+ concentration is slightly higher than the extracellular one and the plasma-to-cell chemical gradient for Na+ is weak, though even the molal concentrations may differ significantly. Consistent with the high intracellular Na+ and low K+ concentrations, a very low or no ouabain-sensitive Na+,K+-ATPase activity and no K+-activated pNPPase activity were found in the plasma membrane fraction from red cells. The Cl- and Mg2+ concentrations expressed per liter cell water were significantly higher in red cells than in plasma whereas the opposite was the case with Ca2+. The distribution of Cl- thus does not seem compatible with an inside-negative membrane potential in mink erythrocytes. In spite of a steep calcium gradient across the red cell membrane, neither a calmodulin-activated Ca2+-ATPase activity nor an ATP-activated Ca2+-pNPPase activity were detectable in the plasma membrane fraction. The origin of a supposed primary Ca2+ gradient for sustaining of osmotic balance thus seems uncertain.

  17. Evaluation of stiffness and plastic deformation of active ceramic self-ligating bracket clips after repetitive opening and closure movements.

    Science.gov (United States)

    Carneiro, Grace Kelly Martins; Roque, Juliano Alves; Segundo, Aguinaldo Silva Garcez; Suzuki, Hideo

    2015-01-01

    The aim of this study was to assess whether repetitive opening and closure of self-ligating bracket clips can cause plastic deformation of the clip. Three types of active/interactive ceramic self-ligating brackets (n = 20) were tested: In-Ovation C, Quicklear and WOW. A standardized controlled device performed 500 cycles of opening and closure movements of the bracket clip with proper instruments and techniques adapted as recommended by the manufacturer of each bracket type. Two tensile tests, one before and one after the repetitive cycles, were performed to assess the stiffness of the clips. To this end, a custom-made stainless steel 0.40 x 0.40 mm wire was inserted into the bracket slot and adapted to the universal testing machine (EMIC DL2000), after which measurements were recorded. On the loading portion of the loading-unloading curve of clips, the slope fitted a first-degree equation curve to determine the stiffness/deflection rate of the clip. The results of plastic deformation showed no significant difference among bracket types before and after the 500 cycles of opening and closure (p = 0.811). There were significant differences on stiffness among the three types of brackets (p = 0.005). The WOW bracket had higher mean values, whereas Quicklear bracket had lower values, regardless of the opening/closure cycle. Repetitive controlled opening and closure movements of the clip did not alter stiffness or cause plastic deformation.

  18. Comparison of the Modeling Approach between Membrane Bioreactor and Conventional Activated Sludge Processes

    DEFF Research Database (Denmark)

    Jiang, Tao; Sin, Gürkan; Spanjers, Henri

    2009-01-01

    Activated sludge models (ASM) have been developed and largely applied in conventional activated sludge (CAS) systems. The applicability of ASM to model membrane bioreactors (MBR) and the differences in modeling approaches have not been studied in detail. A laboratory-scale MBR was modeled using ASM......2d. It was found that the ASM2d model structure can still be used for MBR modeling. There are significant differences related to ASM modeling. First, a lower maximum specific growth rate for MBR nitrifiers was estimated. Independent experiments demonstrated that this might be attributed...... to the inhibition effect of soluble microbial products (SMP) at elevated concentration. Second, a greater biomass affinity to oxygen and ammonium was found, which was probably related to smaller MBR sludge flocs. Finally, the membrane throughput during membrane backwashing/relaxation can be normalized...

  19. Influence of membrane potential on conductance sublevels of chloride channels activated by GABA.

    Science.gov (United States)

    Gage, P W; Chung, S H

    1994-02-22

    Single-channel chloride currents activated by 0.5 microM GABA were recorded in cell-attached and inside-out membrane patches from rat cultured hippocampal neurons. The currents displayed multiple conductance states and outward rectification. The number and amplitude of conductance levels were determined over a range of potentials by using digital signal-processing techniques. It was found that, except for a level close to zero, subconductance levels were regularly spaced. There were fewer sublevels at hyperpolarized than at depolarized potentials, and the spacing between levels varied linearly with potential giving an incremental conductance of 8-10 pS that was independent of membrane potential. Outward rectification is related to the change in the number of conductance levels with potential. One hypothesis that is consistent with these observations is that a channel is composed of a number of synchronized, non-rectifying, conducting pores, and that the number of pores activated changes with membrane potential.

  20. Src-mediated caveolin-1 phosphorylation affects the targeting of active Src to specific membrane sites

    Science.gov (United States)

    Gottlieb-Abraham, Efrat; Shvartsman, Dmitry E.; Donaldson, John C.; Ehrlich, Marcelo; Gutman, Orit; Martin, G. Steven; Henis, Yoav I.

    2013-01-01

    Src interactions with the plasma membrane are an important determinant of its activity. In turn, Src activity modulates its association with the membrane through binding of activated Src to phosphotyrosylated proteins. Caveolin-1 (Cav-1), a major component of caveolae, is a known Src phosphorylation target, and both were reported to regulate cell transformation. However, the nature of Src-Cav-1 interactions, a potential mechanism of their coregulation, remained unclear. Here we used fluorescence recovery after photobleaching beam-size analysis, coimmunoprecipitation, quantitative imaging, and far-Western studies with cells expressing wild type, as well as structural and activity mutants of Src–green fluorescent protein and Cav-1–monomeric red fluorescent protein, to measure their interactions with the membrane and with each other. We show dynamic Src–plasma membrane interactions, which are augmented and stabilized by Cav-1. The mechanism involves phosphorylation of Cav-1 at Tyr-14 by Src and subsequent binding of the Src SH2 domain to phospho–Cav-1, leading to accumulation of activated Src in focal adhesions. This novel Cav-1 function potentially modulates focal adhesion dynamics. PMID:24131997

  1. Voltage-dependent activation in purified reconstituted sodium channels from rabbit T-tubular membranes

    Energy Technology Data Exchange (ETDEWEB)

    Furman, R.E.; Tanaka, J.C.; Mueller, P.; Barchi, R.L.

    1986-01-01

    The authors have examined the voltage-dependent gating of batrachotoxin-modified sodium channels purified from rabbit T-tubular membranes in two ways. First, purified channels were reconstituted into planar bilayers and single-channel properties were measured. Batrachotoxin-activated channels showed steep voltage-dependent activation with half-maximal opening probabilities at potentials between -95 and -116 mV. The single-channel conductance averaged 20 pS and was independent of membrane potential. A second approach was used to establish that this voltage dependence was a characteristic of the entire population of purified channels and not just those few channels observed in planar bilayers. Channels reconstituted into egg phosphophatidyl-choline vesicles were functionally oriented by inclusion of internal saxitoxin; vesicle membrane potentials were then generated by K/sup +/ gradients in the presence of valinomycin. All of the specific /sup 22/Na/sup +/ influx activated by batrachotoxin and blocked by saxitoxin was found to be voltage sensitive, activating between predicted membrane potentials of -100 and -50 mV. The single-channel properties of the purified T-tubular sodium channel correspond closely to those seen with native sodium channels from rat sarcolemma. The voltage-dependent activation of the bactrachotoxin-modified reconstituted channel is the same as that seen with native channels in situ or in bilayers after exposure this toxin.

  2. Erythrocyte membrane ATPase and calcium pumping activities in porcine malignant hyperthermia

    International Nuclear Information System (INIS)

    Thatte, H.S.; Mickelson, J.R.; Addis, P.B.; Louis, C.F.

    1987-01-01

    To investigate possible abnormalities in erythrocyte membrane enzyme activities in the pharmacogenetic disorder MH, membrane ATPase activities have been examined in erythrocyte ghosts prepared from red blood cells of MHS and normal swine. While no differences were noted in Mg2+-ATPase activities, the (Na+, K+)-ATPase activity of MHS erythrocyte ghosts was less than that of normal ghosts. Ca2+-ATPase activity exhibited low- and high-affinity Ca2+-binding sites in both types of erythrocyte ghost. While the Km for Ca2+ was greater for normal than for MHS erythrocyte ghosts at the high-affinity Ca2+-binding site, the reverse was true at the low-affinity Ca2+-binding site. Irrespective of the type of calcium binding site occupied, the Vmax for normal erythrocyte ghost Ca2+-ATPase activity was greater than that for MHS ghosts. In the presence of calmodulin, there was now no difference between MHS and normal erythrocyte ghosts in either the Km for Ca2+ or the Vmax of the Ca2+-ATPase activity. To determine if the calcium pumping activity of intact MHS and normal pig erythrocytes differed, calcium efflux from the 45 Ca-loaded erythrocytes was determined; this activity was significantly greater for MHS than for normal erythrocytes. Thus, the present study confirms that there are abnormalities in the membranes of MHS pig red blood cells. However, we conclude that these abnormalities are unlikely to result in an impaired ability of MHS erythrocytes to regulate their cytosolic Ca2+ concentration

  3. Correlating antimicrobial activity and model membrane leakage induced by nylon-3 polymers and detergents.

    Science.gov (United States)

    Hovakeemian, Sara G; Liu, Runhui; Gellman, Samuel H; Heerklotz, Heiko

    2015-09-14

    Most antimicrobial peptides act upon target microorganisms by permeabilizing their membranes. The mode of action is often assessed by vesicle leakage experiments that use model membranes, with the assumption that biological activity correlates with the permeabilization of the lipid bilayer. The current work aims to extend the interpretation of vesicle leakage results and examine the correlation between vesicle leakage and antimicrobial activity. To this end, we used a lifetime-based leakage assay with calcein-loaded vesicles to study the membrane permeabilizing properties of a novel antifungal polymer poly-NM, two of its analogs, and a series of detergents. In conjunction, the biological activities of these compounds against Candida albicans were assessed and correlated with data from vesicle leakage. Poly-NM induces all-or-none leakage in polar yeast lipid vesicles at the polymer's MIC, 3 μg mL(-1). At this and higher concentrations, complete leakage after an initial lag time was observed. Concerted activity tests imply that this polymer acts independently of the detergent octyl glucoside (OG) for both vesicle leakage and activity against C. albicans spheroplasts. In addition, poly-NM was found to have negligible activity against zwitterionic vesicles and red blood cells. Our results provide a consistent, detailed picture of the mode of action of poly-NM: this polymer induces membrane leakage by electrostatic lipid clustering. In contrast, poly-MM:CO, a nylon-3 polymer comprised of both cationic and hydrophobic segments, seems to act by a different mechanism that involves membrane asymmetry stress. Vesicle leakage for this polymer is transient (limited to nylon-3 polymers we examined act via similar mechanisms; it is surprising that their mechanisms are so distinct. Some, but not all mechanisms of vesicle permeabilization allow for antimicrobial activity.

  4. Bactericidal activity of curcumin I is associated with damaging of bacterial membrane.

    Directory of Open Access Journals (Sweden)

    Poonam Tyagi

    Full Text Available Curcumin, an important constituent of turmeric, is known for various biological activities, primarily due to its antioxidant mechanism. The present study focused on the antibacterial activity of curcumin I, a significant component of commercial curcumin, against four genera of bacteria, including those that are Gram-positive (Staphylococcus aureus and Enterococcus faecalis and Gram-negative (Escherichia coli and Pseudomonas aeruginosa. These represent prominent human pathogens, particularly in hospital settings. Our study shows the strong antibacterial potential of curcumin I against all the tested bacteria from Gram-positive as well as Gram-negative groups. The integrity of the bacterial membrane was checked using two differential permeabilization indicating fluorescent probes, namely, propidium iodide and calcein. Both the membrane permeabilization assays confirmed membrane leakage in Gram-negative and Gram-positive bacteria on exposure to curcumin I. In addition, scanning electron microscopy and fluorescence microscopy were employed to confirm the membrane damages in bacterial cells on exposure to curcumin I. The present study confirms the broad-spectrum antibacterial nature of curcumin I, and its membrane damaging property. Findings from this study could provide impetus for further research on curcumin I regarding its antibiotic potential against rapidly emerging bacterial pathogens.

  5. Cholesterol-Enriched Domain Formation Induced by Viral-Encoded, Membrane-Active Amphipathic Peptide.

    Science.gov (United States)

    Hanson, Joshua M; Gettel, Douglas L; Tabaei, Seyed R; Jackman, Joshua; Kim, Min Chul; Sasaki, Darryl Y; Groves, Jay T; Liedberg, Bo; Cho, Nam-Joon; Parikh, Atul N

    2016-01-05

    The α-helical (AH) domain of the hepatitis C virus nonstructural protein NS5A, anchored at the cytoplasmic leaflet of the endoplasmic reticulum, plays a role in viral replication. However, the peptides derived from this domain also exhibit remarkably broad-spectrum virocidal activity, raising questions about their modes of membrane association. Here, using giant lipid vesicles, we show that the AH peptide discriminates between membrane compositions. In cholesterol-containing membranes, peptide binding induces microdomain formation. By contrast, cholesterol-depleted membranes undergo global softening at elevated peptide concentrations. Furthermore, in mixed populations, the presence of ∼100 nm vesicles of viral dimensions suppresses these peptide-induced perturbations in giant unilamellar vesicles, suggesting size-dependent membrane association. These synergistic composition- and size-dependent interactions explain, in part, how the AH domain might on the one hand segregate molecules needed for viral assembly and on the other hand furnish peptides that exhibit broad-spectrum virocidal activity. Copyright © 2016 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  6. Atmospheric-pressure plasma activation and surface characterization on polyethylene membrane separator

    Science.gov (United States)

    Tseng, Yu-Chien; Li, Hsiao-Ling; Huang, Chun

    2017-01-01

    The surface hydrophilic activation of a polyethylene membrane separator was achieved using an atmospheric-pressure plasma jet. The surface of the atmospheric-pressure-plasma-treated membrane separator was found to be highly hydrophilic realized by adjusting the plasma power input. The variations in membrane separator chemical structure were confirmed by Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy. Chemical analysis showed newly formed carbonyl-containing groups and high surface concentrations of oxygen-containing species on the atmospheric-pressure-plasma-treated polymeric separator surface. It also showed that surface hydrophilicity primarily increased from the polar component after atmospheric-pressure plasma treatment. The surface and pore structures of the polyethylene membrane separator were examined by scanning electron microscopy, revealing a slight alteration in the pore structure. As a result of the incorporation of polar functionalities by atmospheric-pressure plasma activation, the electrolyte uptake and electrochemical impedance of the atmospheric-pressure-plasma-treated membrane separator improved. The investigational results show that the separator surface can be controlled by atmospheric-pressure plasma surface treatment to tailor the hydrophilicity and enhance the electrochemical performance of lithium ion batteries.

  7. Group B streptococcus activates transcriptomic pathways related to premature birth in human extraplacental membranes in vitro.

    Science.gov (United States)

    Park, Hae-Ryung; Harris, Sean M; Boldenow, Erica; McEachin, Richard C; Sartor, Maureen; Chames, Mark; Loch-Caruso, Rita

    2018-03-01

    Streptococcus agalactiae (group B streptococcus [GBS]) infection in pregnant women is the leading cause of infectious neonatal morbidity and mortality in the United States. Although inflammation during infection has been associated with preterm birth, the contribution of GBS to preterm birth is less certain. Moreover, the early mechanisms by which GBS interacts with the gestational tissue to affect adverse pregnancy outcomes are poorly understood. We hypothesized that short-term GBS inoculation activates pathways related to inflammation and premature birth in human extraplacental membranes. We tested this hypothesis using GBS-inoculated human extraplacental membranes in vitro. In agreement with our hypothesis, a microarray-based transcriptomics analysis of gene expression changes in GBS-inoculated membranes revealed that GBS activated pathways related to inflammation and preterm birth with significant gene expression changes occurring as early as 4 h postinoculation. In addition, pathways related to DNA replication and repair were downregulated with GBS treatment. Conclusions based on our transcriptomics data were further supported by responses of prostaglandin E2 (PGE2), and matrix metalloproteinases 1 (MMP1) and 3 (MMP3), all of which are known to be involved in parturition and premature rupture of membranes. These results support our initial hypothesis and provide new information on molecular targets of GBS infection in human extraplacental membranes.

  8. Allosteric activation of membrane-bound glutamate receptors using coordination chemistry within living cells

    Science.gov (United States)

    Kiyonaka, Shigeki; Kubota, Ryou; Michibata, Yukiko; Sakakura, Masayoshi; Takahashi, Hideo; Numata, Tomohiro; Inoue, Ryuji; Yuzaki, Michisuke; Hamachi, Itaru

    2016-10-01

    The controlled activation of proteins in living cells is an important goal in protein-design research, but to introduce an artificial activation switch into membrane proteins through rational design is a significant challenge because of the structural and functional complexity of such proteins. Here we report the allosteric activation of two types of membrane-bound neurotransmitter receptors, the ion-channel type and the G-protein-coupled glutamate receptors, using coordination chemistry in living cells. The high programmability of coordination chemistry enabled two His mutations, which act as an artificial allosteric site, to be semirationally incorporated in the vicinity of the ligand-binding pockets. Binding of Pd(2,2‧-bipyridine) at the allosteric site enabled the active conformations of the glutamate receptors to be stabilized. Using this approach, we were able to activate selectively a mutant glutamate receptor in live neurons, which initiated a subsequent signal-transduction pathway.

  9. Plasma membrane recruitment of dephosphorylated beta-catenin upon activation of the Wnt pathway

    NARCIS (Netherlands)

    Hendriksen, Jolita; Jansen, Marnix; Brown, Carolyn M.; van der Velde, Hella; van Ham, Marco; Galjart, Niels; Offerhaus, G. Johan; Fagotto, Francois; Fornerod, Maarten

    2008-01-01

    The standard model of Wnt signaling specifies that after receipt of a Wnt ligand at the membranous receptor complex, downstream mediators inhibit a cytoplasmic destruction complex, allowing beta-catenin to accumulate in the cytosol and nucleus and co-activate Wnt target genes. Unexpectedly, shortly

  10. Active Pedicle Epithelial Flap Transposition Combined with Amniotic Membrane Transplantation for Treatment of Nonhealing Corneal Ulcers

    OpenAIRE

    Zhang, Ting; Wang, Yuexin; Jia, Yanni; Liu, Dongle; Li, Suxia; Shi, Weiyun; Gao, Hua

    2016-01-01

    Introduction. The objective was to evaluate the efficacy of active pedicle epithelial flap transposition combined with amniotic membrane transplantation (AMT) in treating nonhealing corneal ulcers. Material and Methods. Eleven patients (11 eyes) with nonhealing corneal ulcer who underwent the combined surgery were included. Postoperatively, ulcer healing time was detected by corneal fluorescein staining. Visual acuity, intraocular pressure, surgical complications, and recurrence were recorded...

  11. The relationship between active deformation, stratigraphic architecture, and submarine slope failure in the Santa Barbara Channel, southern California

    Science.gov (United States)

    Wright, A.; Brothers, D. S.; Kluesner, J.; Dugan, B.; Johnson, S. Y.

    2017-12-01

    Multiple submarine landslides are documented along the slope of the seismically active Santa Barbara Basin and are proposed to have been capable of generating tsunamis with 2 - 20 m wave heights. Questions remain as to what factors preconditioned sections of the continental slope for failure and whether adjacent slopes are at risk for future failure. For example, the Gaviota landslide (0.01 - 0.02 km3) and Goleta landslide complex (3 - 4 km3) are surrounded by sections of intact slope sediment, but show signs of incipient failure, emphasizing the need for detailed hazard assessment of the region. Past studies have had to rely on legacy 2D seismic reflection datasets to analyze the regional framework geology and these data did not provide the necessary resolution to identity active faults and folds beneath the slope. Here we integrate 3D multichannel seismic (MCS) reflection volumes (acquired in 1995) as well as high-resolution 2D MCS and multibeam bathymetry datasets (acquired in 2016) to analyze the relationships between slope failure and near-surface deformation, stratigraphic architecture, and substrate fluid flow. The new high-resolution data reveal evidence for active deformation along a previously undocumented network of en echelon faults striking 25° counterclockwise to the dominant structural grain of the region. We observe a locally uplifted zone beneath the Gaviota slide that evidently deforms 90 ka sediment. Analogous patterns of deformation are identified beneath adjacent sections of intact slope, suggesting these areas may be preconditioned for failure. Thinning and onlap of fluid-saturated basin strata occurs above zones of active uplift and fault-propagation folding. We present a conceptual model for differential compaction that induces a lateral pressure gradient and updip migration of fluids, which creates a mid-slope zone of overpressure (i.e., reduced shear strength) along the axis of folding and explains the positioning of the Gaviota slide and

  12. Mapping membrane activity in undiscovered peptide sequence space using machine learning.

    Science.gov (United States)

    Lee, Ernest Y; Fulan, Benjamin M; Wong, Gerard C L; Ferguson, Andrew L

    2016-11-29

    There are some ∼1,100 known antimicrobial peptides (AMPs), which permeabilize microbial membranes but have diverse sequences. Here, we develop a support vector machine (SVM)-based classifier to investigate ⍺-helical AMPs and the interrelated nature of their functional commonality and sequence homology. SVM is used to search the undiscovered peptide sequence space and identify Pareto-optimal candidates that simultaneously maximize the distance σ from the SVM hyperplane (thus maximize its "antimicrobialness") and its ⍺-helicity, but minimize mutational distance to known AMPs. By calibrating SVM machine learning results with killing assays and small-angle X-ray scattering (SAXS), we find that the SVM metric σ correlates not with a peptide's minimum inhibitory concentration (MIC), but rather its ability to generate negative Gaussian membrane curvature. This surprising result provides a topological basis for membrane activity common to AMPs. Moreover, we highlight an important distinction between the maximal recognizability of a sequence to a trained AMP classifier (its ability to generate membrane curvature) and its maximal antimicrobial efficacy. As mutational distances are increased from known AMPs, we find AMP-like sequences that are increasingly difficult for nature to discover via simple mutation. Using the sequence map as a discovery tool, we find a unexpectedly diverse taxonomy of sequences that are just as membrane-active as known AMPs, but with a broad range of primary functions distinct from AMP functions, including endogenous neuropeptides, viral fusion proteins, topogenic peptides, and amyloids. The SVM classifier is useful as a general detector of membrane activity in peptide sequences.

  13. Active liquid treatment by a combination of precipitation and membrane processes

    International Nuclear Information System (INIS)

    Gutman, R.G.; Cumming, I.W.; Williams, G.H.

    1986-08-01

    New ultrafiltration processes developed for the treatment of low and medium active radioactive wastes, were applied successfully to a variety of simulated and real wastes, including magnesium alloy clad spent storage fuel pond waters, reprocessing plant solvent wash liquors, plutonium production effluents and mixed site effluents. After initial laboratory scale feasibility experiments the process was scaled up successfully, using a variety of different ultrafiltration modules. The information accumulated on membrane performance, membrane fouling and flux restoration techniques, and ancillary equipment performance was used to design a much larger demonstration pilot plant. This plant has been constructed and is now processing continuously each day over 1m 3 of a real radioactive effluent. (author)

  14. Hsp70 translocates into the plasma membrane after stress and is released into the extracellular environment in a membrane-associated form that activates macrophages.

    Science.gov (United States)

    Vega, Virginia L; Rodríguez-Silva, Monica; Frey, Tiffany; Gehrmann, Mathias; Diaz, Juan Carlos; Steinem, Claudia; Multhoff, Gabriele; Arispe, Nelson; De Maio, Antonio

    2008-03-15

    Heat shock proteins (hsps) are intracellular chaperones that play a key role in the recovery from stress. Hsp70, the major stress-induced hsp, has been found in the extracellular medium and is capable of activating immune cells. The mechanism involved in Hsp70 release is controversial because this protein does not present a consensual secretory signal. In this study, we have shown that Hsp70 integrates into artificial lipid bilayer openings of ion conductance pathways. In addition, this protein was found inserted into the plasma membrane of cells after stress. Hsp70 was released into the extracellular environment in a membrane-associated form, sharing the characteristics of this protein in the plasma membrane. Extracellular membranes containing Hsp70 were at least 260-fold more effective than free recombinant protein in inducing TNF-alpha production as an indicator of macrophage activation. These observations suggest that Hsp70 translocates into the plasma membrane after stress and is released within membranous structures from intact cells, which could act as a danger signal to activate the immune system.

  15. Synthetic activity enhancement of membrane-bound lipase from Rhizopus chinensis by pretreatment with isooctane.

    Science.gov (United States)

    Wang, Dong; Xu, Yan; Teng, Yun

    2007-05-01

    The cell-bound lipase from Rhizopus chinensis CCTCC M201021 with high catalysis ability for ester synthesis was located as a membrane-bound lipase by the treatments of Yatalase firstly. In order to improve its synthetic activity in non-aqueous phase, the pretreatments of this enzyme with various organic solvents were investigated. The pretreatment with isooctane improved evidently the lipase synthetic activity, resulting in about 139% in relative synthetic activity and 115% in activity recovery. The morphological changes of mycelia caused by organic solvent pretreatments could influence the exposure of the membrane-bound enzyme from mycelia and the exhibition of the lipase activity. The pretreatment conditions with isooctane and acetone were further investigated, and the optimum effect was obtained by the isooctane pretreatment at 4 degrees C for 1 h, resulting in 156% in relative synthetic activity and 126% in activity recovery. When the pretreated lipases were employed as catalysts for the esterification production of ethyl hexanoate in heptane, higher initial reaction rate and higher final molar conversion were obtained using the lipase pretreated with isooctane, compared with the untreated lyophilized one. This result suggested that the pretreatment of the membrane-bound lipase with isooctane could be an effective method to substitute the lyophilization for preparing biocatalysts used in non-aqueous phase reactions.

  16. In vivo antibacterial activity and pharmacological properties of the membrane-active glycopeptide antibiotic YV11455.

    Science.gov (United States)

    Yarlagadda, Venkateswarlu; Konai, Mohini M; Manjunath, Goutham B; Prakash, Relekar G; Mani, Bhuvana; Paramanandham, Krishnamoorthy; Ranjan, Shome B; Ravikumar, Raju; Chakraborty, Subhankari P; Roy, Somenath; Haldar, Jayanta

    2015-06-01

    The membrane-active glycopeptide antibiotic YV11455 is a lipophilic cationic vancomycin analogue that demonstrates rapid and concentration-dependent killing of clinically relevant multidrug-resistant (MDR) Gram-positive bacteria in vitro. YV11455 was 2-fold and 54-270-fold more effective than vancomycin against clinical isolates of vancomycin-sensitive and vancomycin-resistant bacteria, respectively. In this study, the in vivo efficacy, pharmacodynamics, pharmacokinetics and acute toxicology of YV11455 were investigated. In vivo activity and pharmacodynamics were determined in the neutropenic mouse thigh infection model against meticillin-resistant Staphylococcus aureus (MRSA). YV11455 produced dose-dependent reductions in MRSA titres in thigh muscle. When administered intravenously, the 50% effective dose (ED(50)) for YV11455 against MRSA was found to be 3.3 mg/kg body weight, and titres were reduced by up to ca. 3log(10)CFU/g from pre-treatment values at a dosage of 12 mg/kg with single treatment. Single-dose pharmacokinetic studies demonstrated linear kinetics and a prolonged half-life, with an increase in drug exposure (area under the concentration-time curve) compared with vancomycin. The peak plasma concentration following an intravenous dose of 12 mg/kg was 543.5 μg/mL. Acute toxicology studies revealed that YV11455 did not cause any significant alterations in biochemical parameters or histological pictures related to major organs such as the liver and kidney at its pharmacodynamic endpoint (ED(3-log kill)). These findings collectively suggest that YV11455 could be used clinically for the treatment of infections caused by MDR Gram-positive bacteria. Copyright © 2015 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  17. Growth hormone activates phospholipase C in proximal tubular basolateral membranes from canine kidney

    International Nuclear Information System (INIS)

    Rogers, S.A.; Hammerman, M.R.

    1989-01-01

    To delineate pathways for signal transduction by growth hormone (GH) in proximal tubule, the authors incubated basolateral membranes isolated from canine kidney with human growth hormone (hGH) or human prolactin (hPrl) and measured levels of inositol trisphosphate (InsP 3 ) in suspensions and of diacylglycerol extractable from the membranes. Incubation with hGH, but not hPrl, increased levels of InsP 3 and diacylglycerol in a concentration-dependent manner. Half-maximal effects occurred between 0.1 and 1 nM hGH. Increased levels of InsP 3 were measured after as little as 5 sec of incubation with 1 nM hGH, and increase was maximal after 15 sec. Increases were no longer detectable after 60 sec because of dephosphorylation of InsP 3 in membrane suspensions. hGH did not affect rates of dephosphorylation. hGH-stimulated increases in InsP 3 were detectable in membranes suspended in 0, 0.1, and 0.2 μM calcium but not in 0.3 or 1.0 μM calcium. 125 I-labeled hGH-receptor complexes with M r values of 66,000 and 140,000 were identified in isolated basolateral membranes. The findings establish that GH activates phospholipase C in isolated canine renal proximal tubular basolateral membranes, potentially after binding to a specific receptor. This process could mediate signal transmission by GH across the plasma membrane of the proximal tubular cell and elsewhere

  18. Antimycotic activity of fengycin C biosurfactant and its interaction with phosphatidylcholine model membranes.

    Science.gov (United States)

    González-Jaramillo, Lina María; Aranda, Francisco José; Teruel, José Antonio; Villegas-Escobar, Valeska; Ortiz, Antonio

    2017-08-01

    Lipopeptide biosurfactants constitute one of the most promising groups of compounds for the treatment and prevention of fungal diseases in plants. Bacillus subtilis strain EA-CB0015 produces iturin A, fengycin C and surfactin and it has been proven useful for the treatment of black Sigatoka disease in banana plants, an important pathology caused by the fungus Mycosphaerella fijiensis (Morelet). We have found that B. subtilis EA-CB0015 cell free supernatants and purified fractions inhibit M. fijiensis cellular growth. The effect of the purified lipopeptides mentioned above on fungal growth has been also evaluated, observing that iturin A and fengycin C inhibit mycelial growth and ascospore germination, whereas surfactin is not effective. On the hypothesis that the antifungal action of the lipopeptides is associated to their incorporation into biological membranes, ultimately leading to membrane permeabilization, a detailed biophysical study on the interaction of a new isoform of fengycin C with model dipalmitoyphosphatidylcholine (DPPC) membranes has been carried out. Differential scanning calorimetry shows that fengycin C alters the thermotropic phase transitions of DPPC, and is laterally segregated in the fluid bilayer forming domains. Fluorescent probe polarization measurements show that fengycin C does not affect the hydrophobic interior of the membrane. This latter perturbation is concomitant with a strong dehydration of the polar region of DPPC, as shown by FTIR. Fengycin-rich domains, where the surrounding DPPC molecules are highly dehydrated, may well constitute sites of membrane permeabilization leading to a leaky target membrane. These results are a solid support to explain the membrane perturbing action of fengycin, which has been related to its antifungal activity. Copyright © 2017 Elsevier B.V. All rights reserved.

  19. Numerical simulation and experimental validation of the large deformation bending and folding behavior of magneto-active elastomer composites

    International Nuclear Information System (INIS)

    Sheridan, Robert; VonLockette, Paris R; Roche, Juan; Lofland, Samuel E

    2014-01-01

    This work seeks to provide a framework for the numerical simulation of magneto-active elastomer (MAE) composite structures for use in origami engineering applications. The emerging field of origami engineering employs folding techniques, an array of crease patterns traditionally on a single flat sheet of paper, to produce structures and devices that perform useful engineering operations. Effective means of numerical simulation offer an efficient way to optimize the crease patterns while coupling to the performance and behavior of the active material. The MAE materials used herein are comprised of nominally 30% v/v, 325 mesh barium hexafarrite particles embedded in Dow HS II silicone elastomer compound. These particulate composites are cured in a magnetic field to produce magneto-elastic solids with anisotropic magnetization, e.g. they have a preferred magnetic axis parallel to the curing axis. The deformed shape and/or blocked force characteristics of these MAEs are examined in three geometries: a monolithic cantilever as well as two- and four-segment composite accordion structures. In the accordion structures, patches of MAE material are bonded to a Gelest OE41 unfilled silicone elastomer substrate. Two methods of simulation, one using the Maxwell stress tensor applied as a traction boundary condition and another employing a minimum energy kinematic (MEK) model, are investigated. Both methods capture actuation due to magnetic torque mechanisms that dominate MAE behavior. Comparison with experimental data show good agreement with only a single adjustable parameter, either an effective constant magnetization of the MAE material in the finite element models (at small and moderate deformations) or an effective modulus in the minimum energy model. The four-segment finite element model was prone to numerical locking at large deformation. The effective magnetization and modulus values required are a fraction of the actual experimentally measured values which suggests a

  20. Numerical simulation and experimental validation of the large deformation bending and folding behavior of magneto-active elastomer composites

    Science.gov (United States)

    Sheridan, Robert; Roche, Juan; Lofland, Samuel E.; vonLockette, Paris R.

    2014-09-01

    This work seeks to provide a framework for the numerical simulation of magneto-active elastomer (MAE) composite structures for use in origami engineering applications. The emerging field of origami engineering employs folding techniques, an array of crease patterns traditionally on a single flat sheet of paper, to produce structures and devices that perform useful engineering operations. Effective means of numerical simulation offer an efficient way to optimize the crease patterns while coupling to the performance and behavior of the active material. The MAE materials used herein are comprised of nominally 30% v/v, 325 mesh barium hexafarrite particles embedded in Dow HS II silicone elastomer compound. These particulate composites are cured in a magnetic field to produce magneto-elastic solids with anisotropic magnetization, e.g. they have a preferred magnetic axis parallel to the curing axis. The deformed shape and/or blocked force characteristics of these MAEs are examined in three geometries: a monolithic cantilever as well as two- and four-segment composite accordion structures. In the accordion structures, patches of MAE material are bonded to a Gelest OE41 unfilled silicone elastomer substrate. Two methods of simulation, one using the Maxwell stress tensor applied as a traction boundary condition and another employing a minimum energy kinematic (MEK) model, are investigated. Both methods capture actuation due to magnetic torque mechanisms that dominate MAE behavior. Comparison with experimental data show good agreement with only a single adjustable parameter, either an effective constant magnetization of the MAE material in the finite element models (at small and moderate deformations) or an effective modulus in the minimum energy model. The four-segment finite element model was prone to numerical locking at large deformation. The effective magnetization and modulus values required are a fraction of the actual experimentally measured values which suggests a

  1. Sphingolipids activate membrane fusion of Semliki Forest virus in a stereospecific manner

    DEFF Research Database (Denmark)

    Moesby, Lise; Corver, J; Erukulla, R K

    1995-01-01

    on degradation of the viral capsid protein by trypsin encapsulated in the target liposomes. Fusion mediated by D-erythro-ceramide was not affected by the additional presence in the target liposomes of ceramide stereoisomers incapable of fusion activation. Binding of the virus to the liposomes, as assessed...... by flotation on sucrose density gradients, was not dependent on the presence of fusion-competent or fusion-incompetent sphingolipids in the liposomes. The results of this study support the notion that a stereospecific interaction of the viral fusion protein with D-erythro sphingolipids in the target membrane......The alphavirus Semliki Forest virus (SFV) enters cells through receptor-mediated endocytosis. Subsequently, triggered by the acid pH in endosomes, the viral envelope fuses with the endosomal membrane. Membrane fusion of SFV has been shown previously to be dependent on the presence of cholesterol...

  2. Improved ZIF-8 membrane: Effect of activation procedure and determination of diffusivities of light hydrocarbons

    KAUST Repository

    Pan, Yichang

    2015-06-23

    Zeolitic imidazolate framework ZIF-8 has shown great potential for effective separation of hydrocarbon mixtures based on its intrinsic ultramicroporous feature. In order to explore the permeation and diffusion properties of hydrocarbons through ZIF-8 membrane, high-quality ZIF-8 membranes with a separation factor of ~90 for propylene/propane are successfully prepared via optimizing the activation processes. Single-component permeation data for hydrocarbons (C1–C4) through the improved ZIF-8 membrane are measured and analyzed by Maxwell-Stefan (MS) model to get the transport diffusivities of these hydrocarbons. The diffusivity values of hydrocarbon compare well with those obtained by other experimental techniques. Binary mixture permeation also can be well predicted through single-component adsorption parameters.

  3. The effect of charged lipids on bacteriorhodopsin membrane reconstitution and its photochemical activities

    International Nuclear Information System (INIS)

    Wang Zhen; Bai Jing; Xu Yuhong

    2008-01-01

    Bacteriorhodopsin (BR) was reconstituted into artificial lipid membrane containing various charged lipid compositions. The proton pumping activity of BR under flash and continuous illumination, proton permeability across membrane, as well as the decay kinetics of the photocycle intermediate M 412 were studied. The results showed that lipid charges would significantly affect the orientation of BR inserted into lipid membranes. In liposomes containing anionic lipids, BRs were more likely to take natural orientation as in living cells. In neutral or positively charged liposomes, most BRs were reversely assembled, assuming an inside out orientation. Moreover, the lipids charges also affect BR's M intermediate kinetics, especially the slow component in M intermediate decay. The half-life M 412s increased significantly in BRs in liposomes containing cationic lipids, while decreased in those in anionic liposomes

  4. Block copolymer hollow fiber membranes with catalytic activity and pH-response

    KAUST Repository

    Hilke, Roland

    2013-08-14

    We fabricated block copolymer hollow fiber membranes with self-assembled, shell-side, uniform pore structures. The fibers in these membranes combined pores able to respond to pH and acting as chemical gates that opened above pH 4, and catalytic activity, achieved by the incorporation of gold nanoparticles. We used a dry/wet spinning process to produce the asymmetric hollow fibers and determined the conditions under which the hollow fibers were optimized to create the desired pore morphology and the necessary mechanical stability. To induce ordered micelle assembly in the doped solution, we identified an ideal solvent mixture as confirmed by small-angle X-ray scattering. We then reduced p-nitrophenol with a gold-loaded fiber to confirm the catalytic performance of the membranes. © 2013 American Chemical Society.

  5. Characterization of membrane determinant in old T-cells with suppressor activity

    International Nuclear Information System (INIS)

    Hendricks, L.C.; Heidrick, M.L.

    1986-01-01

    T-cell function declines with age. Many T-cell functions are initiated at the cell membrane; therefore, age-related membrane alterations may contribute to loss of function. They have previously reported developing a monoclonal antibody, HH-AGE-T(1), which recognizes a cell with suppressor activity and binds to 15-20% of the T-cells from old BC3F 1 mice, but only to 0-4% of young T-cells. To further characterize the determinant recognized by HH-AGE-T(1), they analyzed immunoprecipitates (IP) of young and old T-cell membranes by 2D-SDS PAGE, followed by Western blotting. Immunodetection of the blots showed that HH-AGE-T(1) bound a heterodimer (66 kD, pI 8.44 and 36 kD, pI 5.82-7.12 subunits) in IP from old mice; but not young mice. Monoclonal anti-Lyt 2 antibody did not bind the determinant. When IP of iodinated T-cells were run on SDS-PAGE gels followed by blotting and autoradiography of the blots, very prominent bands were detected in the old sample and faint bands were detected in the young sample. These results suggest that HH-AGE-T(1) recognizes a membrane protein which is present in small amounts on young T-cells but which increases markedly with age. Further studies are needed to determine the significance of this age-related membrane change

  6. Isolating active orogenic wedge deformation in the southern Subandes of Bolivia

    Science.gov (United States)

    Weiss, Jonathan R.; Brooks, Benjamin A.; Foster, James H.; Bevis, Michael; Echalar, Arturo; Caccamise, Dana; Heck, Jacob; Kendrick, Eric; Ahlgren, Kevin; Raleigh, David; Smalley, Robert; Vergani, Gustavo

    2016-08-01

    A new GPS-derived surface velocity field for the central Andean backarc permits an assessment of orogenic wedge deformation across the southern Subandes of Bolivia, where recent studies suggest that great earthquakes (>Mw 8) are possible. We find that the backarc is not isolated from the main plate boundary seismic cycle. Rather, signals from subduction zone earthquakes contaminate the velocity field at distances greater than 800 km from the Chile trench. Two new wedge-crossing velocity profiles, corrected for seasonal and earthquake affects, reveal distinct regions that reflect (1) locking of the main plate boundary across the high Andes, (2) the location of and loading rate at the back of orogenic wedge, and (3) an east flank velocity gradient indicative of décollement locking beneath the Subandes. Modeling of the Subandean portions of the profiles indicates along-strike variations in the décollement locked width (WL) and wedge loading rate; the northern wedge décollement has a WL of ~100 km while accumulating slip at a rate of ~14 mm/yr, whereas the southern wedge has a WL of ~61 km and a slip rate of ~7 mm/yr. When compared to Quaternary estimates of geologic shortening and evidence for Holocene internal wedge deformation, the new GPS-derived wedge loading rates may indicate that the southern wedge is experiencing a phase of thickening via reactivation of preexisting internal structures. In contrast, we suspect that the northern wedge is undergoing an accretion or widening phase primarily via slip on relatively young thrust-front faults.

  7. Transferred nuclear Overhauser effect analyses of membrane-bound enkephalin analogues by sup 1 H nuclear magnetic resonance: Correlation between activities and membrane-bound conformations

    Energy Technology Data Exchange (ETDEWEB)

    Milon, Alain; Miyazawa, Tatsuo; Higashijima, Tsutomu (Univ. of Tokyo (Japan))

    1990-01-09

    Leu-enkephalin, (D-Ala{sup 2})Leu-enkephalin, and (D-Ala{sup 2})Leu-enkephalinamide (agonists) and (L-Ala{sup 2})Leu-enkephalin (inactive analogue) bind to lipid bilayer consisting of phosphatidylcholine and phosphatidylserine. The conformations that these compounds assume, once bound to perdeuterated phospholipid bilayer, have been shown to be unique, as shown by the transferred nuclear Overhauser effect (TRNOE) of {sup 1}H NMR spectroscopy. In addition, their location in the bilayer was analyzed by TRNOE in the presence of spin-labeled phospholipids. These analyses showed a clear relationship between the activity and the peptide-membrane interaction. The three active peptides, when bound to membranes, adopt the same conformation, characterized by a type II{prime} {beta}-turn around Gly{sup 3}-Phe and a {gamma}-turn around Gly{sup 2} (or D-Ala{sup 2}). The inactive analogue, (L-Ala{sup 2})Leu-enkephalin, displayed a completely different TRNOE pattern corresponding to a different conformation in the membrane-bound state. The tyrosine residue of the active compounds is not inserted into the interior of membrane, but it is inserted into the bilayer for the L-Ala{sup 2} analogue. According to these results, (L-Ala{sup 2})Leu-enkephalin may be explained to be inactive because the mode of binding to the membranes is different from that of active compounds.

  8. Direct Cytoskeleton Forces Cause Membrane Softening in Red Blood Cells

    Science.gov (United States)

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

    2015-01-01

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

  9. Shape-dependent bactericidal activity of copper oxide nanoparticle mediated by DNA and membrane damage

    Energy Technology Data Exchange (ETDEWEB)

    Laha, Dipranjan; Pramanik, Arindam [Department of Life Science and Biotechnology, Jadavpur University, 188, Raja S C Mallick Road, Kolkata 700032 (India); Laskar, Aparna [CSIR-Indian Institute of Chemical Biology, Kolkata 700032 (India); Jana, Madhurya [Department of Life Science and Biotechnology, Jadavpur University, 188, Raja S C Mallick Road, Kolkata 700032 (India); Pramanik, Panchanan [Department of Chemistry, Indian Institute of Technology, Kharagpur 721302 (India); Karmakar, Parimal, E-mail: pkarmakar_28@yahoo.co.in [Department of Life Science and Biotechnology, Jadavpur University, 188, Raja S C Mallick Road, Kolkata 700032 (India)

    2014-11-15

    Highlights: • Spherical and sheet shaped copper oxide nanoparticles were synthesized. • Physical characterizations of these nanoparticles were done by TEM, DLS, XRD, FTIR. • They showed shape dependent antibacterial activity on different bacterial strain. • They induced both membrane damage and ROS mediated DNA damage in bacteria. - Abstract: In this work, we synthesized spherical and sheet shaped copper oxide nanoparticles and their physical characterizations were done by the X-ray diffraction, fourier transform infrared spectroscopy, transmission electron microscopy and dynamic light scattering. The antibacterial activity of these nanoparticles was determined on both gram positive and gram negative bacterial. Spherical shaped copper oxide nanoparticles showed more antibacterial property on gram positive bacteria where as sheet shaped copper oxide nanoparticles are more active on gram negative bacteria. We also demonstrated that copper oxide nanoparticles produced reactive oxygen species in both gram negative and gram positive bacteria. Furthermore, they induced membrane damage as determined by atomic force microscopy and scanning electron microscopy. Thus production of and membrane damage are major mechanisms of the bactericidal activity of these copper oxide nanoparticles. Finally it was concluded that antibacterial activity of nanoparticles depend on physicochemical properties of copper oxide nanoparticles and bacterial strain.

  10. Shape-dependent bactericidal activity of copper oxide nanoparticle mediated by DNA and membrane damage

    International Nuclear Information System (INIS)

    Laha, Dipranjan; Pramanik, Arindam; Laskar, Aparna; Jana, Madhurya; Pramanik, Panchanan; Karmakar, Parimal

    2014-01-01

    Highlights: • Spherical and sheet shaped copper oxide nanoparticles were synthesized. • Physical characterizations of these nanoparticles were done by TEM, DLS, XRD, FTIR. • They showed shape dependent antibacterial activity on different bacterial strain. • They induced both membrane damage and ROS mediated DNA damage in bacteria. - Abstract: In this work, we synthesized spherical and sheet shaped copper oxide nanoparticles and their physical characterizations were done by the X-ray diffraction, fourier transform infrared spectroscopy, transmission electron microscopy and dynamic light scattering. The antibacterial activity of these nanoparticles was determined on both gram positive and gram negative bacterial. Spherical shaped copper oxide nanoparticles showed more antibacterial property on gram positive bacteria where as sheet shaped copper oxide nanoparticles are more active on gram negative bacteria. We also demonstrated that copper oxide nanoparticles produced reactive oxygen species in both gram negative and gram positive bacteria. Furthermore, they induced membrane damage as determined by atomic force microscopy and scanning electron microscopy. Thus production of and membrane damage are major mechanisms of the bactericidal activity of these copper oxide nanoparticles. Finally it was concluded that antibacterial activity of nanoparticles depend on physicochemical properties of copper oxide nanoparticles and bacterial strain

  11. Removal of diatrizoate with catalytically active membranes incorporating microbially produced palladium nanoparticles.

    Science.gov (United States)

    Hennebel, Tom; De Corte, Simon; Vanhaecke, Lynn; Vanherck, Katrien; Forrez, Ilse; De Gusseme, Bart; Verhagen, Pieter; Verbeken, Kim; Van der Bruggen, Bart; Vankelecom, Ivo; Boon, Nico; Verstraete, Willy

    2010-03-01

    There is an increasing concern about the fate of iodinated contrast media (ICM) in the environment. Limited removal efficiencies of currently applied techniques such as advanced oxidation processes require more performant strategies. The aim of this study was to establish an innovative degradation process for diatrizoate, a highly recalcitrant ICM, by using biogenic Pd nanoparticles as free suspension or immobilized in polyvinylidene fluoride (PVDF) and polysulfone (PSf) membranes. As measured by HPLC-UV, the removal of 20mg L(-1) diatrizoate by a 10mg L(-1) Pd suspension was completed after 4h at a pH of 10. LC-MS analysis provided evidence for the sequential hydrodeiodination of diatrizoate. Pd did not lose its activity after incorporation in the PVDF and PSf matrix and the highest activity (k(cat)=30.0+/-0.4h(-1) L g(-1) Pd) was obtained with a casting solution of 10% PSf and 500mg L(-1) Pd. Subsequently, water containing 20mg L(-1) diatrizoate was treated in a membrane contactor, in which the water was supplied at one side of the membrane while hydrogen was provided at the other side. In a fed batch configuration, a removal efficiency of 77% after a time period of 48h was obtained. This work showed that membrane contactors with encapsulated biogenic nanoparticles can be instrumental for treatment of water contaminated with diatrizoate. Copyright 2009 Elsevier Ltd. All rights reserved.

  12. Effect of Powdered Activated Carbon to Reduce Fouling in Membrane Bioreactors: A Sustainable Solution. Case Study

    Directory of Open Access Journals (Sweden)

    Giuseppe Mancini

    2013-04-01

    Full Text Available Membrane Bio Reactors (MBRs are mainly used for industrial wastewaters applications where their costs can be more easily afforded. High costs are basically due to energy consumption and membrane cleaning or replacement. Membrane fouling is responsible for reducing treated water production and increasing maintenance as well as operation costs. According to previous researches, the addition of Powdered Activated Carbon (PAC in high dosages could reduce membrane fouling; but such concentrations are economically unsustainable for operative conditions. A MBR pilot plant, fed by mixed liquor of a full-scale activated sludge process from a municipal wastewater treatment plant, was operated dosing low PAC concentrations (0, 2, 5, 10 and 20 mg·L−1, respectively. Experiments were also carried out at two different temperatures corresponding to summer and winter conditions. Results indicated that PAC addition was effective at the low dosages (2 and 5 mg·L−1 by reducing the permeate flux loss (from 16 up to 27%, respectively while higher PAC concentrations turns out in a useless cost increase.

  13. Thyroid hormones increase Na -H exchange activity in renal brush border membranes

    Energy Technology Data Exchange (ETDEWEB)

    Kinsella, J.; Sacktor, B.

    1985-06-01

    Na -H exchange activity, i.e., amiloride-sensitive Na and H flux, in renal proximal tubule brush border (luminal) membrane vesicles was increased in the hyperthyroid rat and decreased in the hypothyroid rat, relative to the euthyroid animal. A positive correlation was found between Na -H exchange activity and serum concentrations of thyroxine (T4) and triiodothyronine (T3). The thyroid status of the animal did not alter amiloride-insensitive Na uptake. The rate of passive pH gradient dissipation was higher in membrane vesicles from hyperthyroid rats compared to the rate in vesicles from hypothyroid animals, a result which would tend to limit the increase in Na uptake in vesicles from hyperthyroid animals. Na -dependent phosphate uptake was increased in membrane vesicles from hyperthyroid rats; Na -dependent D-glucose and L-proline uptakes were not changed by the thyroid status of the animal. The effect of thyroid hormones in increasing the uptake of Na in the brush border membrane vesicle is consistent with the action of the hormones in enhancing renal Na reabsorption.

  14. Laser-activated membrane introduction mass spectrometry for high-throughput evaluation of bulk heterogeneous catalysts.

    Science.gov (United States)

    Nayar, Amit; Liu, Renxuan; Allen, Robert J; McCall, Michael J; Willis, Richard R; Smotkin, Eugene S

    2002-05-01

    Laser-activated membrane introduction mass spectrometry (LAMIMS), a high-throughput screening method, evaluates heterogeneous catalysts under realistic reactor conditions. It is a precise, versatile system requiring no moving parts. The catalyst array is supported on carbon paper overlaid upon a silicone rubber membrane configured in a variation of membrane introduction mass spectrometry as introduced by Cooks. The carbon paper serves as a heat-dissipating gas diffusion layer that permits laser heating of catalyst samples far above the decomposition temperature of the polymer membrane that separates the array from the mass spectrometer vacuum chamber. A computer-controlled CO2 bar code writing laser is used for fine-tune heating of the catalyst spots above the base temperature of the LAMIMS reactor. The detailed design and performance of LAMIMS is demonstrated on arrays of "real world" bulk water-gas shift catalysts using natural and isotopically labeled reactor feed streams. A bulk catalyst array spot can be evaluated for activity and selectivity in as little as 1.5 min. All array screening results were confirmed by industrial microreactor evaluations.

  15. Photodynamic activity of the boronated chlorin e6 amide in artificial and cellular membranes.

    Science.gov (United States)

    Antonenko, Yuri N; Kotova, Elena A; Omarova, Elena O; Rokitskaya, Tatyana I; Ol'shevskaya, Valentina A; Kalinin, Valery N; Nikitina, Roza G; Osipchuk, Julia S; Kaplan, Mikhail A; Ramonova, Alla A; Moisenovich, Mikhail M; Agapov, Igor I; Kirpichnikov, Mikhail P

    2014-03-01

    Photodynamic tumor-destroying activity of the boronated chlorin e6 derivative BACE (chlorin e6 13(1)-N-{2-[N-(1-carba-closo-dodecaboran-1-yl)methyl]aminoethyl}amide-15(2), 17(3)-dimethyl ester), previously described in Moisenovich et al. (2010) PLoS ONE 5(9) e12717, was shown here to be enormously higher than that of unsubstituted chlorin e6, being supported by the data on much higher photocytotoxicity of BACE in M-1 sarcoma cell culture. To validate membrane damaging effect as the basis of the enhanced tumoricidal activity, BACE was compared with unsubstituted chlorin e6 in the potency to photosensitize dye leakage from liposomes, transbilayer lipid flip-flop, inactivation of gramicidin A ionic channels in planar lipid membranes and erythrocyte hemolysis. In all the models comprising artificial and cellular membranes, the photodynamic effect of BACE exceeded that of chlorin e6. BACE substantially differed from chlorin e6 in the affinity to liposomes and erythrocytes, as monitored by fluorescence spectroscopy, flow cytometry and centrifugation. The results support the key role of membrane binding in the photodynamic effect of the boronated chlorin e6 amide. Copyright © 2013 Elsevier B.V. All rights reserved.

  16. What determines the activity of antimicrobial and cytolytic peptides in model membranes.

    Science.gov (United States)

    Clark, Kim S; Svetlovics, James; McKeown, Alesia N; Huskins, Laura; Almeida, Paulo F

    2011-09-20

    We previously proposed three hypotheses relating the mechanism of antimicrobial and cytolytic peptides in model membranes to the Gibbs free energies of binding and insertion into the membrane [Almeida, P. F., and Pokorny, A. (2009) Biochemistry 48, 8083-8093]. Two sets of peptides were designed to test those hypotheses, by mutating of the sequences of δ-lysin, cecropin A, and magainin 2. Peptide binding and activity were measured on phosphatidylcholine membranes. In the first set, the peptide charge was changed by mutating basic to acidic residues or vice versa, but the amino acid sequence was not altered much otherwise. The type of dye release changed from graded to all-or-none according to prediction. However, location of charged residues in the sequence with the correct spacing to form salt bridges failed to improve binding. In the second set, the charged and other key residues were kept in the same positions, whereas most of the sequence was significantly but conservatively simplified, maintaining the same hydrophobicity and amphipathicity. This set behaved completely different from predicted. The type of release, which was expected to be maintained, changed dramatically from all-or-none to graded in the mutants of cecropin and magainin. Finally, contrary to the hypotheses, the results indicate that the Gibbs energy of binding to the membrane, not the Gibbs energy of insertion, is the primary determinant of peptide activity. © 2011 American Chemical Society

  17. Observing active deformation of volcanoes in North America: Geodetic data from the Plate Boundary Observatory and associated networks

    Science.gov (United States)

    Puskas, C. M.; Phillips, D. A.; Mattioli, G. S.; Meertens, C. M.; Hodgkinson, K. M.; Crosby, C. J.; Enders, M.; Feaux, K.; Mencin, D.; Baker, S.; Lisowski, M.; Smith, R. B.

    2013-12-01

    The EarthScope Plate Boundary Observatory (PBO), operated by UNAVCO, records deformation of the geologically diverse North America western plate boundary, with subnetworks of instruments concentrated at selected active and potentially active volcanoes. These sensors record deformation and earthquakes and allow monitoring agencies and researchers to analyze changes in ground motion and seismicity. The intraplate volcanoes at Yellowstone and Long Valley are characterized by uplift/subsidence cycles, high seismicity, and hydrothermal activity but there have been no historic eruptions at either volcano. PBO maintains dense GPS networks of 20-25 stations at each of these volcanoes, with an additional 5 boreholes at Yellowstone containing tensor strainmeters, short-period seismometers, and borehole tiltmeters. Subduction zone volcanoes in the Aleutian Arc have had multiple historic eruptions, and PBO maintains equipment at Augustine (8 GPS), Akutan (8 GPS, 4 tiltmeters), and Unimak Island (14 GPS, 8 tiltmeters). The Unimak stations are at the active Westdahl and Shishaldin edifices and the nearby, inactive Isanotski volcano. In the Cascade Arc, PBO maintains networks at Mount St. Helens (15 GPS, 4 borehole strainmeters and seismometers, 8 borehole tiltmeters), Shasta (7 GPS, 1 borehole strainmeter and seismometer), and Lassen Peak (8 GPS). Data from many of these stations in the Pacific Northwest and California are also provided as realtime streams of raw and processed data. Real-time GPS data, along with high-rate GPS data, will be an important new resource for detecting and studying future rapid volcanic deformation events and earthquakes. UNAVCO works closely with the USGS Volcano Hazards Program, archiving data from USGS GPS stations in Alaska, Cascadia, and Long Valley. The PBO and USGS networks combined provide more comprehensive coverage than PBO alone, particularly of the Cascade Arc, where the USGS maintains a multiple instruments near each volcano. Ground

  18. Evaluation of stiffness and plastic deformation of active ceramic self-ligating bracket clips after repetitive opening and closure movements

    Directory of Open Access Journals (Sweden)

    Grace Kelly Martins Carneiro

    2015-08-01

    Full Text Available OBJECTIVE: The aim of this study was to assess whether repetitive opening and closure of self-ligating bracket clips can cause plastic deformation of the clip.METHODS: Three types of active/interactive ceramic self-ligating brackets (n = 20 were tested: In-Ovation C, Quicklear and WOW. A standardized controlled device performed 500 cycles of opening and closure movements of the bracket clip with proper instruments and techniques adapted as recommended by the manufacturer of each bracket type. Two tensile tests, one before and one after the repetitive cycles, were performed to assess the stiffness of the clips. To this end, a custom-made stainless steel 0.40 x 0.40 mm wire was inserted into the bracket slot and adapted to the universal testing machine (EMIC DL2000, after which measurements were recorded. On the loading portion of the loading-unloading curve of clips, the slope fitted a first-degree equation curve to determine the stiffness/deflection rate of the clip.RESULTS: The results of plastic deformation showed no significant difference among bracket types before and after the 500 cycles of opening and closure (p = 0.811. There were significant differences on stiffness among the three types of brackets (p = 0.005. The WOW bracket had higher mean values, whereas Quicklear bracket had lower values, regardless of the opening/closure cycle.CONCLUSION: Repetitive controlled opening and closure movements of the clip did not alter stiffness or cause plastic deformation.

  19. The activation energy for insertion of transmembrane alpha-helices is dependent on membrane composition.

    Science.gov (United States)

    Meijberg, Wim; Booth, Paula J

    2002-06-07

    The physical mechanisms that govern the folding and assembly of integral membrane proteins are poorly understood. It appears that certain properties of the lipid bilayer affect membrane protein folding in vitro, either by modulating helix insertion or packing. In order to begin to understand the origin of this effect, we investigate the effect of lipid forces on the insertion of a transmembrane alpha-helix using a water-soluble, alanine-based peptide, KKAAAIAAAAAIAAWAAIAAAKKKK-amide. This peptide binds to preformed 1,2-dioleoyl-l-alpha-phosphatidylcholine (DOPC) vesicles at neutral pH, but spontaneous transmembrane helix insertion directly from the aqueous phase only occurs at high pH when the Lys residues are de-protonated. These results suggest that the translocation of charge is a major determinant of the activation energy for insertion. Time-resolved measurements of the insertion process at high pH indicate biphasic kinetics with time constants of ca 30 and 430 seconds. The slower phase seems to correlate with formation of a predominantly transmembrane alpha-helical conformation, as determined from the transfer of the tryptophan residue to the hydrocarbon region of the membrane. Temperature-dependent measurements showed that insertion can proceed only above a certain threshold temperature and that the Arrhenius activation energy is of the order of 90 kJ mol(-1). The kinetics, threshold temperature and the activation energy change with the mole fraction of 1,2-dioleoyl-l-alpha-phosphatidylethanolamine (DOPE) introduced into the DOPC membrane. The activation energy increases with increasing DOPE content, which could reflect the fact that this lipid drives the bilayer towards a non-bilayer transition and increases the lateral pressure in the lipid chain region. This suggests that folding events involving the insertion of helical segments across the bilayer can be controlled by lipid forces. (c) 2002 Elsevier Science Ltd.

  20. Delta-Opioid receptors exhibit high efficiency when activating trimeric G proteins in membrane domains

    Czech Academy of Sciences Publication Activity Database

    Bouřová, Lenka; Koštrnová, Alexandra; Hejnová, Lucie; Moravcová, Zuzana; Moon, H. E.; Novotný, Jiří; Milligan, G.; Svoboda, Petr

    2003-01-01

    Roč. 85, č. 1 (2003), s. 34-49 ISSN 0022-3042 R&D Projects: GA MŠk LN00A026; GA ČR GA309/01/0255 Grant - others:Welcome Trust(GB) xx Institutional research plan: CEZ:AV0Z5011922 Keywords : membrane domains * GTPase activity * G protein coupling Subject RIV: CE - Biochemistry Impact factor: 4.825, year: 2003

  1. Auxin-activated NADH oxidase activity of soybean plasma membranes is distinct from the constitutive plasma membrane NADH oxidase and exhibits prion-like properties

    Science.gov (United States)

    Morre, D. James; Morre, Dorothy M.; Ternes, Philipp

    2003-01-01

    The hormone-stimulated and growth-related cell surface hydroquinone (NADH) oxidase activity of etiolated hypocotyls of soybeans oscillates with a period of about 24 min or 60 times per 24-h day. Plasma membranes of soybean hypocotyls contain two such NADH oxidase activities that have been resolved by purification on concanavalin A columns. One in the apparent molecular weight range of 14-17 kDa is stimulated by the auxin herbicide 2,4-dichlorophenoxyacetic acid (2,4-D). The other is larger and unaffected by 2,4-D. The 2,4-D-stimulated activity absolutely requires 2,4-D for activity and exhibits a period length of about 24 min. Also exhibiting 24-min oscillations is the rate of cell enlargement induced by the addition of 2,4-D or the natural auxin indole-3-acetic acid (IAA). Immediately following 2,4-D or IAA addition, a very complex pattern of oscillations is frequently observed. However, after several hours a dominant 24-min period emerges at the expense of the constitutive activity. A recruitment process analogous to that exhibited by prions is postulated to explain this behavior.

  2. Investigating Sterol and Redox Regulation of the Ion Channel Activity of CLIC1 Using Tethered Bilayer Membranes

    Directory of Open Access Journals (Sweden)

    Heba Al Khamici

    2016-12-01

    Full Text Available The Chloride Intracellular Ion Channel (CLIC family consists of six conserved proteins in humans. These are a group of enigmatic proteins, which adopt both a soluble and membrane bound form. CLIC1 was found to be a metamorphic protein, where under specific environmental triggers it adopts more than one stable reversible soluble structural conformation. CLIC1 was found to spontaneously insert into cell membranes and form chloride ion channels. However, factors that control the structural transition of CLIC1 from being an aqueous soluble protein into a membrane bound protein have yet to be adequately described. Using tethered bilayer lipid membranes and electrical impedance spectroscopy system, herein we demonstrate that CLIC1 ion channel activity is dependent on the type and concentration of sterols in bilayer membranes. These findings suggest that membrane sterols play an essential role in CLIC1’s acrobatic switching from a globular soluble form to an integral membrane form, promoting greater ion channel conductance in membranes. What remains unclear is the precise nature of this regulation involving membrane sterols and ultimately determining CLIC1’s membrane structure and function as an ion channel. Furthermore, our impedance spectroscopy results obtained using CLIC1 mutants, suggest that the residue Cys24 is not essential for CLIC1’s ion channel function. However Cys24 does appear important for optimal ion channel activity. We also observe differences in conductance between CLIC1 reduced and oxidized forms when added to our tethered membranes. Therefore, we conclude that both membrane sterols and redox play a role in the ion channel activity of CLIC1.

  3. Nanoparticle-Mediated Mechanical Destruction of Cell Membranes: A Coarse-Grained Molecular Dynamics Study.

    Science.gov (United States)

    Zhang, Liuyang; Zhao, Yiping; Wang, Xianqiao

    2017-08-16

    The effects of binding mode, shape, binding strength, and rotational speed of actively rotating nanoparticles on the integrity of cell membranes have been systematically studied using dissipative particle dynamics simulations. With theoretical analyses of lipid density, surface tension, stress distribution, and water permeation, we demonstrate that the rotation of nanoparticles can provide a strong driving force for membrane rupture. The results show that nanoparticles embedded inside a cell membrane via endocytosis are more capable of producing large membrane deformations under active rotation than nanoparticles attached on the cell membrane surface. Nanoparticles with anisotropic shapes produce larger deformation and have a higher rupture efficiency than those with symmetric shapes. Our findings provide useful design guidelines for a general strategy based on utilizing mechanical forces to rupture cell membranes and therefore destroy the integrity of cells.

  4. Studies on the membrane-associated proteolytic activities of Escherichia coli

    International Nuclear Information System (INIS)

    Palmer, S.M.

    1986-01-01

    The membrane fraction contains three proteolytic activities which can be resolved from whole membrane detergent extracts by DEAE-cellulose chromatography. The first two eluting activities have been previously reported as protease V and protease IV. These two enzymes were further purified by gel permeation HPLC. Protease V has a M. W. of 31,000 in SDS-PAGE gels. Protease IV has a M. W. of 62,000 and exists in two distinct isoforms of pl ≅ 6.7 and 6.9. The third enzyme eluting from the DEAE-cellulose column was further purified by affinity chromatography on Benzamidine-Sepharose 6B. This enzyme, referred to herein as protease VI, is a 43 kdal protein which has not been previously characterized. Protease VI was sensitive to inhibition by the serine protease inhibitors phenylmethylsulfony fluoride (PMSF), diisopropylfluorophosphate (DFP) and p-amino-benzamidine (PAB). Additionally, this enzyme exhibited maximal activity at pH ≅ 8.0. Incubation of whole membrane preparations with [ 3 H] DFP resulted in 11 specific proteins acquiring the radioactive label, included in this group of proteins were proteases IV, V, and VI. Several of the DFP-reactive proteins were also shown to bind [ 125 I] ampicillin

  5. Xenoestrogen-Induced ERK-1 and ERK-2 Activation via Multiple Membrane-Initiated Signaling Pathways

    Science.gov (United States)

    Bulayeva, Nataliya N.; Watson, Cheryl S.

    2004-01-01

    Xenoestrogens can mimic or antagonize the activity of physiological estrogens, and the suggested mechanism of xenoestrogen action involves binding to estrogen receptors (ERs). However, the failure of various in vitro or in vivo assays to show strong genomic activity of xenoestrogens compared with estradiol (E2) makes it difficult to explain their ability to cause abnormalities in animal (and perhaps human) reproductive functions via this pathway of steroid action. E2 has also been shown to initiate rapid intracellular signaling, such as changes in levels of intracellular calcium, cAMP, and nitric oxide, and activations of a variety of kinases, via action at the membrane. In this study, we demonstrate that several xenoestrogens can rapidly activate extracellular-regulated kinases (ERKs) in the pituitary tumor cell line GH3/B6/F10, which expresses high levels of the membrane receptor for ER-α(mER). We tested a phytoestrogen (coumestrol), organochlorine pesticides or their metabolites (endosulfan, dieldrin, and DDE), and detergent by-products of plastics manufacturing (p-nonylphenol and bisphenol A). These xenoestrogens (except bisphenol A) produced rapid (3–30 min after application), concentration (10−14–10−8 M)-dependent ERK-1/2 phosphorylation but with distinctly different activation patterns. To identify signaling pathways involved in ERK activation, we used specific inhibitors of ERs, epidermal growth factor receptors, Ca2+ signaling, Src and phosphoinositide-3 kinases, and a membrane structure disruption agent. Multiple inhibitors blocked ERK activation, suggesting simultaneous use of multiple pathways and complex signaling web interactions. However, inhibitors differentially affected each xenoestrogen response examined. These actions may help to explain the distinct abilities of xenoestrogens to disrupt reproductive functions at low concentrations. PMID:15531431

  6. Activation of stretch-activated channels and maxi-K+ channels by membrane stress of human lamina cribrosa cells.

    LENUS (Irish Health Repository)

    Irnaten, Mustapha

    2009-01-01

    The lamina cribrosa (LC) region of the optic nerve head is considered the primary site of damage in glaucomatous optic neuropathy. Resident LC cells have a profibrotic potential when exposed to cyclical stretch. However, the mechanosensitive mechanisms of these cells remain unknown. Here the authors investigated the effects of membrane stretch on cell volume change and ion channel activity and examined the associated changes in intracellular calcium ([Ca(2+)](i)).

  7. A Geodynamic Study of Active Crustal Deformation and Earthquakes in North China

    Science.gov (United States)

    Yang, Y.; Liu, M.

    2005-12-01

    North China is part of the Archaean Sino-Korean craton, yet today it is a region of intense crustal deformation and earthquakes, including 21 M >=7.0 events since 512 AD. More than half of the large events occurred within the Fen-Wei rift system surrounding the stable Ordos plateau; the largest events (M >=7.3) show a sequential southward migration along the rift. However, since 1695 the Fen-Wei rift has became seismically dormant, while seismicity seems having shifted eastward to the North China plain, marked by the 1996 Tangshan earthquake (M=7.8). We have developed a 3D viscoelastic geodynamic model to study the cause of seismicity and its spatial-temporal pattern in North China. Constrained by crustal kinematics from GPS and neotectonic data, the model shows high deviatoric stress in the North China crust, resulting mainly from compression of the expanding Tibetan Plateau and resistance from the stable Siberian block. Within North China seismicity is largely controlled by lateral heterogeneity of lithospheric structures, which explains the concentration of seismicity in the Fen-Wei rift. Our results show that stress triggering may have contributed to the sequential migration of large events along the rift, and the release and migration of stress and strain energy from these large events may partially explain the intense seismicity in the North China plain in the past 300 years. Comparing the predicted long-term spatial pattern of strain energy with seismic energy release provides some insights of potential earthquake risks in North China.

  8. Regulation of the membrane insertion and conductance activity of the metamorphic chloride intracellular channel protein CLIC1 by cholesterol.

    Directory of Open Access Journals (Sweden)

    Stella M Valenzuela

    Full Text Available The Chloride Intracellular ion channel protein CLIC1 has the ability to spontaneously insert into lipid membranes from a soluble, globular state. The precise mechanism of how this occurs and what regulates this insertion is still largely unknown, although factors such as pH and redox environment are known contributors. In the current study, we demonstrate that the presence and concentration of cholesterol in the membrane regulates the spontaneous insertion of CLIC1 into the membrane as well as its ion channel activity. The study employed pressure versus area change measurements of Langmuir lipid monolayer films; and impedance spectroscopy measurements using tethered bilayer membranes to monitor membrane conductance during and following the addition of CLIC1 protein. The observed cholesterol dependent behaviour of CLIC1 is highly reminiscent of the cholesterol-dependent-cytolysin family of bacterial pore-forming proteins, suggesting common regulatory mechanisms for spontaneous protein insertion into the membrane bilayer.

  9. The activation loop of PIP5K functions as a membrane sensor essential for lipid substrate processing

    Science.gov (United States)

    Liu, Aizhuo; Sui, Dexin; Wu, Dianqing; Hu, Jian

    2016-01-01

    Phosphatidylinositol 4-phosphate 5-kinase (PIP5K), a representative member of the phosphatidylinositol phosphate kinase (PIPK) family, is a major enzyme that biosynthesizes the signaling molecule PI(4,5)P2 (phosphatidylinositol 4,5-bisphosphate) in eukaryotic cells. The stringent specificity toward lipid substrates and the high sensitivity to the membrane environment strongly suggest a membrane-sensing mechanism, but the underlying structural basis is still largely unknown. We present a nuclear magnetic resonance (NMR) study on a peptide commensurate with a PIP5K’s activation loop, which has been reported to be a determinant of lipid substrate specificity and subcellular localization of PIP5K. Although the activation loop is severely disordered in the crystal structure of PIP5K, the NMR experiments showed that the largely unstructured peptide folded into an amphipathic helix upon its association with the 1,2-dihexanoyl-sn-glycero-3-phosphocholine (DHPC) micellar surface. Systematic mutagenesis and functional assays further demonstrated the crucial roles of the amphipathic helix and its hydrophobic surface in kinase activity and membrane-sensing function, supporting a working model in which the activation loop is a critical structural module conferring a membrane-sensing mechanism on PIP5K. The activation loop, surprisingly functioning as a membrane sensor, represents a new paradigm of kinase regulation by the activation loop through protein-membrane interaction, which also lays a foundation on the regulation of PIP5K (and other PIPKs) by membrane lipids for future studies. PMID:28138522

  10. Diphytanoyl lipids as model systems for studying membrane-active peptides.

    Science.gov (United States)

    Kara, Sezgin; Afonin, Sergii; Babii, Oleg; Tkachenko, Anton N; Komarov, Igor V; Ulrich, Anne S

    2017-10-01

    The branched chains in diphytanoyl lipids provide membranes with unique properties, such as high chemical/physical stability, low water permeability, and no gel-to-fluid phase transition at ambient temperature. Synthetic diphytanoyl phospholipids are often used as model membranes for electrophysiological experiments. To evaluate whether these sturdy lipids are also suitable for solid-state NMR, we have examined their interactions with a typical amphiphilic peptide in comparison with straight-chain lipids. First, their phase properties were monitored using 31 P NMR, and the structural behaviour of the antimicrobial peptide PGLa was studied by 19 F NMR and circular dichroism in oriented membrane samples. Only lipids with choline headgroups (DPhPC) were found to form stable lipid bilayers in oriented samples, while DPhPG, DPhPE and DPhPS display non-lamellar structures. Hence, the experimental temperature and hydration are crucial factors when using supported diphytanoyl lipids, as both parameters must be maintained in an appropriate range to avoid the formation of non-bilayer structures. For the same reason, a high content of other diphytanoyl lipids besides DPhPC in mixed lipid systems is not favourable. Unlike the situation in straight-chain membranes, we found that the α-helical PGLa was not able to insert into the tightly packed fluid bilayer of DPhPC but remained in a surface-bound state even at very high peptide concentration. This behaviour can be explained by the high cohesivity and the negative spontaneous curvature of the diphytanoyl lipids. These characteristic features must therefore be taken into consideration, both, in electrophysiological studies, and when interpreting the structural behaviour of membrane-active peptides in such lipid environment. Copyright © 2017 Elsevier B.V. All rights reserved.

  11. Effects of Three Kinds of Curcuminoids on Anti-Oxidative System and Membrane Deformation of Human Peripheral Blood Erythrocytes in High Glucose Levels

    Directory of Open Access Journals (Sweden)

    Wei Yang

    2015-01-01

    Full Text Available Background/Aims: Curcuminoids are the main bioactive constituents of the rhizome of turmeric. Erythrocytes lesions in diabetes are probably related to hyperglycemia and protein glycation. It has been reported that curcumin prevent lipid peroxidation. However, reports on the effects of demethoxycurcumin and bis-demethoxycurcumin on human erythrocytes at high glucose levels are scarce. Our aim is to investigate the effect of curcuminoids on oxidative stress and membrane of erythrocytes exposed to hyperglycemic condition. Methods: In this study, the different blood samples were treated with two doses of glucose (10 or 30 mM to mimic hyperglycemia in the presence or absence of three kinds of curcuminoids (5 or 10 μM in a medium at 37 °C for 24 h (Each experiment consists of 20 blood samples from 10 male and 10 female volunteers. The malondialdehyde was checked by HPLC, antioxidase (GSH and GSSG were measured by LC/MS, SOD was checked by WST-1 kit, morphology and phospholipid symmetry were detected by flow cytometry, confocal scanning microscope and scanning electron microscope. Results: The results illustrated that all three curcuminoids reduce oxidative stress damage on the membrane and maintain a better profile for erythrocytes. Furthermore, three curcuminoids had benefit effects on antioxidase. Conclusion: The three kinds of curcuminoids supplementation may prevent lipid peroxidation at different intensity and membrane dysfunction of human erythrocytes in hyperglycemia.

  12. Startup of the Anammox Process in a Membrane Bioreactor (AnMBR) from Conventional Activated Sludge.

    Science.gov (United States)

    Gutwiński, P; Cema, G; Ziembińska-Buczyńska, A; Surmacz-Górska, J; Osadnik, M

    2016-12-01

      In this study, a laboratory-scale anammox process in a membrane bioreactor (AnMBR) was used to startup the anaerobic ammonium oxidation (anammox) process from conventional activated sludge. Stable operation was achieved after 125 days. From that time, nitrogen load was gradually increased. After six months, the average nitrogen removal efficiency exceeded 80%. The highest obtained special anammox activity (SAA) achieved was 0.17 g (-N + -N) (g VSS × d)-1. Fluorescent in situ hybridization also proved the presence of the anammox bacteria, typically a genus of Brocadia anammoxidans and Kuenenia stuttgartiensis.

  13. Antioxidant activities of bambara groundnut (Vigna subterranea) protein hydrolysates and their membrane ultrafiltration fractions.

    Science.gov (United States)

    Arise, Abimbola K; Alashi, Adeola M; Nwachukwu, Ifeanyi D; Ijabadeniyi, Oluwatosin A; Aluko, Rotimi E; Amonsou, Eric O

    2016-05-18

    In this study, the bambara protein isolate (BPI) was digested with three proteases (alcalase, trypsin and pepsin), to produce bambara protein hydrolysates (BPHs). These hydrolysates were passed through ultrafiltration membranes to obtain peptide fractions of different sizes (3 kDa. This is in agreement with the result obtained for the ferric reducing power, metal chelating and hydroxyl radical scavenging activities where higher molecular weight peptides exhibited better activity (p alcalase and trypsin hydrolysates (82%). These findings show the potential use of BPHs and their peptide fraction as antioxidants in reducing food spoilage or management of oxidative stress-related metabolic disorders.

  14. Sweetness-induced activation of membrane dipole potential in STC-1 taste cells.

    Science.gov (United States)

    Chen, Li-Chun; Xie, Ning-Ning; Deng, Shao-Ping

    2016-12-01

    The biological functions of cell membranes strongly influence the binding and transport of molecular species. We developed STC-1 cell line stably expressing the sweet taste receptor (T1R2/T1R3), and explored the possible correlation between sweeteners and membrane dipole potential of STC-1 cells. In this study, sweetener-induced dipole potential activation was elucidated using a fluorescence-based measurement technique, by monitoring the voltage sensitive probe Di-8-ANEPPS using a dual wavelength ratiometric approach. It indicated that the presence of sweeteners resulted in cell membrane dipole potential change, and interaction of artificial sweeteners with taste cells resulted in a greater reduction in potential compared with natural sweeteners. Our work presents a newly developed approach using a fluorescence-based measurement technique to study sweetener-induced dipole potential activation of STC-1 cells. This new approach could be used as a complementary tool to study the function of sweet taste receptors or other GPCRs and helps to understand the basis sweetness mechanism. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Development of adsorptive membranes by confinement of activated biochar into electrospun nanofibers

    Directory of Open Access Journals (Sweden)

    Mehrdad Taheran

    2016-11-01

    Full Text Available Adsorptive membranes have many applications in removal of contaminants, such as heavy metals and organic contaminants from water. Recently, increasing concentrations of pharmaceutically active compounds, especially antibiotics, such as chlortetracycline in water and wastewater sources has raised concerns about their potentially adverse impacts on environment and human health. In this study, a series of polyacrylonitrile (PAN/activated biochar nanofibrous membranes (NFMs with different loadings of biochar (0–2%, w/w were fabricated using electrospinning. The morphology and structure of fabricated membranes was investigated by scanning electron microscopy, Fourier transform infrared and thermogravimetric analysis. The results showed that at 1.5% of biochar loading, the surface area reached the maximum value of 12.4 m2/g and beyond this loading value, agglomeration of particles inhibited fine interaction with nanofibrous matrix. Also, the adsorption tests using chlortetracycline showed that, under environmentally relevant concentrations, the fabricated adsorptive NFMs had a potential for removal of these types of emerging contaminants from water and wastewaters.

  16. Interrogating the activities of conformational deformed enzyme by single-molecule fluorescence-magnetic tweezers microscopy

    Science.gov (United States)

    Guo, Qing; He, Yufan; Lu, H. Peter

    2015-01-01

    Characterizing the impact of fluctuating enzyme conformation on enzymatic activity is critical in understanding the structure–function relationship and enzymatic reaction dynamics. Different from studying enzyme conformations under a denaturing condition, it is highly informative to manipulate the conformation of an enzyme under an enzymatic reaction condition while monitoring the real-time enzymatic activity changes simultaneously. By perturbing conformation of horseradish peroxidase (HRP) molecules using our home-developed single-molecule total internal reflection magnetic tweezers, we successfully manipulated the enzymatic conformation and probed the enzymatic activity changes of HRP in a catalyzed H2O2–amplex red reaction. We also observed a significant tolerance of the enzyme activity to the enzyme conformational perturbation. Our results provide a further understanding of the relation between enzyme behavior and enzymatic conformational fluctuation, enzyme–substrate interactions, enzyme–substrate active complex formation, and protein folding–binding interactions. PMID:26512103

  17. Study on the antioxidant activity and membrane interaction of a multiple antioxidant

    DEFF Research Database (Denmark)

    Jodko-Jodko-Piórecka, Kasia; Klösgen, Beate; Litwinienko, Grzegorz

    Catecholamines, including dopamine, have a role in the transduction of nervous stimuli. Apart from that, they might behave as endogenous phenolic antioxidants protecting the neuronal tissue from deleterious effects of oxidative stress. Results from our preliminary study1 even indicate a synergistic...... effect: an interplay of catecholamines with other molecules seems to enhance their antioxidant activity (e.g. 3,4-dihydroxyphenylalanine - L-DOPA -, and 2,2,5,7,8-pentamethyl-6-hydroxychroman - PMHC -, an analogue of α-tocopherol). Addition of small amounts of catecholamines suppresses or largely...... inhibits the peroxidation of lipids2. The understanding of the mechanism of antioxidant action of catecholamines and their interplay with other antioxidants in lipid membranes requires interdisciplinary research on the kinetics and thermodynamics of antioxidant(s)/membrane interactions. Here we report...

  18. Lipopeptaibol metabolites of tolypocladium geodes: total synthesis, preferred conformation, and membrane activity.

    Science.gov (United States)

    Rainaldi, Mario; Moretto, Alessandro; Peggion, Cristina; Formaggio, Fernando; Mammi, Stefano; Peggion, Evaristo; Galvez, José Antonio; Díaz-de-Villegas, Maria Dolores; Cativiela, Carlos; Toniolo, Claudio

    2003-08-04

    We have synthesized by solution methods and characterized the lipopeptaibol metabolite LP237-F8 extracted from the fungus Tolypocladium geodes and five selected analogues with the Etn-->Aib or Etn-->Nva replacement at position 8 and/or a triple Gln-->Glu(OMe) replacement at positions 5, 6, and 9 (Etn=Calpha-ethylnorvaline, Aib=alpha-aminoisobutyric acid, Nva=norvaline). Conformation analysis, performed by FT-IR absorption, NMR, and CD techniques, strongly supports the view that the six terminally blocked decapeptides are highly helical in solution. Helix topology and amphiphilic character are responsible for their remarkable membrane activity. At position 8 the combination of high hydrophobicity and Calpha tetrasubstitution, as in the Etn-containing LP237-F8 metabolite, has a positive effect on membrane interaction.

  19. Ceramic Membrane combined with Powdered Activated Carbon (PAC) or Coagulation for Treatment of Impaired Quality Waters

    KAUST Repository

    Hamad, Juma Z.

    2013-08-29

    Ceramic membranes (CM) are robust membranes attributed with high production, long life span and stability against critical conditions. While capital costs are high, these are partially offset by lower operation and maintenance costs compared to polymeric membranes. Like any other low-pressure membrane (LPM), CM faces problems of fouling, low removal of organic matter and poor removal of trace organic compounds (TOrCs). Current pretreatment approaches that are mainly based on coagulation and adsorption can remove some organic matter but with a low removal of the biopolymers component which is responsible for fouling. Powdered activated carbon (PAC) accompanied with a LPM maintains good removal of TOrCs. However, enhanced removal of TOrCs to higher level is required. Submicron powdered activated carbon (SPAC), obtained after crushing commercial activated carbon into very fine particle, and novel activated carbon (KCU 6) which is characterized with larger pores and high surface area were employed. A pre-coating approach, which provides intimated contact between PAC and contaminants, was adopted for wastewater and (high DOC) surface water treatment. For seawater, in-line coagulation with iron III chloride was adopted. Both SPAC and KCU 6 showed good removal of biopolymers at a dose of 30 mg/L with > 85 % and 90 %, respectively. A dose of 40 mg/L of SPAC and 30 mg/L KCU 6 pre-coats were successful used in controlling membrane fouling. SPAC is suggested to remove biopolymers by physical means and adsorption while KCU 6 removed biopolymers through adsorption. Both KCU 6 and SPAC attained high removal of TOrCs whereas KCU 6 showed outstanding performance. Out of 29 TOrCs investigated, KCU 6 showed > 87 % TOrCs rejection for 28 compounds. In seawater pretreatment, transparent exopolymer particles (TEP) were found to be an important foulant. TEP promoted both reversible and irreversible fouling. TEP are highly electronegative while alumina CM is positively charged which

  20. A protein chip membrane-capture assay for botulinum neurotoxin activity

    International Nuclear Information System (INIS)

    Marconi, Severine; Ferracci, Geraldine; Berthomieu, Maelys; Kozaki, Shunji; Miquelis, Raymond; Boucraut, Jose; Seagar, Michael

    2008-01-01

    Botulinum neurotoxins A and B (BoNT/A and B) are neuromuscular blocking agents which inhibit neurotransmission by cleaving the intra-cellular presynaptic SNARE proteins SNAP-25 and VAMP2, localized respectively in plasma membrane and synaptic vesicles. These neurotoxins are both dangerous pathogens and powerful therapeutic agents with numerous clinical and cosmetic applications. Consequently there is a need for in vitro assays of their biological activity to screen for potential inhibitors and to replace the widely used in vivo mouse assay. Surface plasmon resonance (SPR) was used to measure membrane vesicle capture by antibodies against SNAP-25 and VAMP2. Substrate cleavage by BoNTs modified capture providing a method to assay toxin activity. Firstly using synaptic vesicles as a substrate, a comparison of the EC 50 s for BoNT/B obtained by SPR, ELISA or flow cytometry indicated similar sensitivity although SPR assays were more rapid. Sonication of brain or neuronal cultures generated plasma membrane fragments with accessible intra-cellular epitopes adapted to measurement of BoNT/A activity. SPR responses were proportional to antigen concentration permitting detection of as little as 4 pM SNAP-25 in crude lysates. BoNT/A activity was assayed using monoclonal antibodies that specifically recognize a SNAP-25 epitope generated by the proteolytic action of the toxin. Incubation of intact primary cultured neurons with BoNT/A yielded an EC 50 of 0.5 pM. The SPR biosensor method was sensitive enough to monitor BoNT/A and B activity in cells cultured in a 96-well format providing an alternative to experimental animals for toxicological assays

  1. Antibacterial activity of polyphenolic fraction of Kombucha against Vibrio cholerae: targeting cell membrane.

    Science.gov (United States)

    Bhattacharya, D; Ghosh, D; Bhattacharya, S; Sarkar, S; Karmakar, P; Koley, H; Gachhui, R

    2018-02-01

    The present study was undertaken to determine the mechanism of antibacterial activity of a polyphenolic fraction, composed of mainly catechin and isorhamnetin, previously isolated from Kombucha, a 14-day fermented beverage of sugared black tea, against the enteropathogen Vibrio cholerae N16961. Bacterial growth was found to be seriously impaired by the polyphenolic fraction in a dose-dependent manner. Scanning Electron Microscopy demonstrated morphological alterations in bacterial cells when exposed to the polyphenolic fraction in a concentration-dependent manner. Permeabilization assays confirmed that the fraction disrupted bacterial membrane integrity in both time- and dose-dependent manners, which were proportional to the production of intracellular reactive oxygen species (ROS). Furthermore, each of the polyphenols catechin and isorhamnetin showed the ability to permeate bacterial cell membranes by generating oxidative stress, thereby suggesting their role in the antibacterial potential of Kombucha. Thus, the basic mechanism of antibacterial activity of the Kombucha polyphenolic fraction against V. cholerae involved bacterial membrane permeabilization and morphological changes, which might be due to the generation of intracellular ROS. To the best of our knowledge, this is the first report on the investigation of antibacterial mechanism of Kombucha, which is mostly attributed to its polyphenolic content. The emergence of multidrug-resistant Vibrio cholerae strains has hindered an efficient anti-Vibrio therapy. This study has demonstrated the membrane damage-mediated antibacterial mechanism of Kombucha, a popular fermented beverage of sugared tea, which is mostly attributed to its polyphenolic content. This study also implies the exploitation of Kombucha as a potential new source of bioactive polyphenols against V. cholerae. © 2017 The Society for Applied Microbiology.

  2. Leveraging electrokinetics for the active control of dendritic fullerene-1 release across a nanochannel membrane

    Science.gov (United States)

    Bruno, Giacomo; Geninatti, Thomas; Hood, R. Lyle; Fine, Daniel; Scorrano, Giovanni; Schmulen, Jeffrey; Hosali, Sharath; Ferrari, Mauro; Grattoni, Alessandro

    2015-03-01

    General adoption of advanced treatment protocols such as chronotherapy will hinge on progress in drug delivery technologies that provide precise temporal control of therapeutic release. Such innovation is also crucial to future medicine approaches such as telemedicine. Here we present a nanofluidic membrane technology capable of achieving active and tunable control of molecular transport through nanofluidic channels. Control was achieved through application of an electric field between two platinum electrodes positioned on either surface of a 5.7 nm nanochannel membrane designed for zero-order drug delivery. Two electrode configurations were tested: laser-cut foils and electron beam deposited thin-films, configurations capable of operating at low voltage (nanotechnology platform to facilitate controlled delivery of molecules and particles has broad applicability to next-generation therapeutics for numerous pathologies, including autoimmune diseases, circadian dysfunction, pain, and stress, among others.General adoption of advanced treatment protocols such as chronotherapy will hinge on progress in drug delivery technologies that provide precise temporal control of therapeutic release. Such innovation is also crucial to future medicine approaches such as telemedicine. Here we present a nanofluidic membrane technology capable of achieving active and tunable control of molecular transport through nanofluidic channels. Control was achieved through application of an electric field between two platinum electrodes positioned on either surface of a 5.7 nm nanochannel membrane designed for zero-order drug delivery. Two electrode configurations were tested: laser-cut foils and electron beam deposited thin-films, configurations capable of operating at low voltage (nanotechnology platform to facilitate controlled delivery of molecules and particles has broad applicability to next-generation therapeutics for numerous pathologies, including autoimmune diseases, circadian

  3. Demarcating SurA Activities Required for Outer Membrane Targeting of Yersinia pseudotuberculosis Adhesins

    Science.gov (United States)

    Obi, Ikenna R.

    2013-01-01

    SurA is a periplasmic protein folding factor involved in chaperoning and trafficking of outer membrane proteins across the Gram-negative bacterial periplasm. In addition, SurA also possesses peptidyl-prolyl cis/trans isomerase activity. We have previously reported that in enteropathogenic Yersinia pseudotuberculosis, SurA is needed for bacterial virulence and envelope integrity. In this study, we investigated the role of SurA in the assembly of important Yersinia adhesins. Using genetic mutation, biochemical characterization, and an in vitro-based bacterial host cell association assay, we confirmed that surface localization of the invasin adhesin is dependent on SurA. As a surA deletion also has some impact on the levels of individual components of the BAM complex in the Yersinia outer membrane, abolished invasin surface assembly could reflect both a direct loss of SurA-dependent periplasmic targeting and a potentially compromised BAM complex assembly platform in the outer membrane. To various degrees, the assembly of two other adhesins, Ail and the pH 6 antigen fibrillum PsaA, also depends on SurA. Consequently, loss of SurA leads to a dramatic reduction in Yersinia attachment to eukaryotic host cells. Genetic complementation of surA deletion mutants indicated a prominent role for SurA chaperone function in outer membrane protein assembly. Significantly, the N terminus of SurA contributed most of this SurA chaperone function. Despite a dominant chaperoning role, it was also evident that SurA isomerization activity did make a modest contribution to this assembly process. PMID:23589578

  4. Soft-sediment deformation in a tectonically active area: The Plio-Pleistocene Zarzal Formation in the Cauca Valley (Western Colombia)

    Science.gov (United States)

    Neuwerth, Ralph; Suter, Fiore; Guzman, Carlos A.; Gorin, Georges E.

    2006-04-01

    seismites in the Zarzal Formation represents corroboration of tectonic activity in this area during the Pleistocene. Earthquakes with a magnitude higher than 5 can be postulated, based upon the proximity of active faults and the types of deformations.

  5. Deformation microstructures

    DEFF Research Database (Denmark)

    Hansen, N.; Huang, X.; Hughes, D.A.

    2004-01-01

    Microstructural characterization and modeling has shown that a variety of metals deformed by different thermomechanical processes follows a general path of grain subdivision, by dislocation boundaries and high angle boundaries. This subdivision has been observed to very small structural scales...

  6. Cdc42p and Rho1p are sequentially activated and mechanistically linked to vacuole membrane fusion

    International Nuclear Information System (INIS)

    Logan, Michael R.; Jones, Lynden; Eitzen, Gary

    2010-01-01

    Small monomeric GTPases act as molecular switches, regulating many biological functions via activation of membrane localized signaling cascades. Activation of their switch function is controlled by GTP binding and hydrolysis. Two Rho GTPases, Cdc42p and Rho1p, are localized to the yeast vacuole where they regulate membrane fusion. Here, we define a method to directly examine vacuole membrane Cdc42p and Rho1p activation based on their affinity to probes derived from effectors. Cdc42p and Rho1p showed unique temporal activation which aligned with distinct subreactions of in vitro vacuole fusion. Cdc42p was rapidly activated in an ATP-independent manner while Rho1p activation was kinetically slower and required ATP. Inhibitors that are known to block vacuole membrane fusion were examined for their effect on Cdc42p and Rho1p activation. Rdi1p, which inhibits the dissociation of GDP from Rho proteins, blocked both Cdc42p and Rho1p activation. Ligands of PI(4,5)P 2 specifically inhibited Rho1p activation while pre-incubation with U73122, which targets Plc1p function, increased Rho1p activation. These results define unique activation mechanisms for Cdc42p and Rho1p, which may be linked to the vacuole membrane fusion mechanism.

  7. Serial plasma concentrations of atrial natriuretic peptide, plasma renin activity, aldosterone, and antidiuretic hormone in neonates on extracorporeal membrane oxygenation.

    NARCIS (Netherlands)

    Semmekrot, B.A.; Pesman, G.J.; Span, P.N.; Sweep, C.G.J.; Heyst, A.F.J. van; Monnens, L.A.H.; Bor, M. van de; Tanke, R.B.; Staak, F.H.J.M. van der

    2002-01-01

    To obtain information on water and salt regulating hormones and volume homeostasis during neonatal extracorporeal membrane oxygenation (ECMO), serial determinations of atrial natriuretic peptide (ANP), plasma renin activity (PRA), aldosterone (Aldo), antidiuretic hormone (ADH), colloid-osmotic

  8. Magnetic enzyme membranes as active elements of electrochemical sensors: specific amino acid enzyme elctrodes.

    Science.gov (United States)

    Calvot, C; Berjonneau, A M; Gellf, G; Thomas, D

    1975-11-15

    The basic principle of the described magnetic enzyme electrodes is a kinetic accumulation of CO2 at the active layer electrode interface. The local pCO2 level is linked to three simultaneous phenomena: substrate diffusion in, enzyme reaction CO2 diffusion out. After a transient state there is a stationary state between the quantity of CO2 produced by the enzyme reaction and the CO2 diffusing from the active membrane to the bulk solution. Continuous determination of free amino acids in biological media is useful in biological processing, fermentation, medicine, pharmaceutical industries and biological research. No methods are presently available for any specific continuous measurement of lysine which is of nutritional importance in protein industrial syntheses; of phenylalanine and tyrosine which have to be monitored in several inborn diseases (phenylketonuria being the most important of them); of arginine and histidine which play a still imperfectly understood part in neurochemistry. The use of decarboxylase bearing membranes as sensors in such measurements could offer several novel advantages: (a) a simple device made of a currently manufactured electrode slightly modified by the use of an enzyme membrane; (b) The absence of any enzymic consumption due to the immobilization and the negligible consumption of substrate during the measurements; (c) The sensitivity which can be sharpened by a systematic study of the membrane parameters; (d) the continuous response of the electrode as long as it is in contact with the substrate solution; (e) the further feasibility as a miniature sensor. The magnetic device introduced allows obviously a convenient use of the enzyme electrode, the active part can be removed and replaced without disturbance for the pCO2 electrode itself. The enzyme electrodes are not only useful at the applied point of view but also at the fundamental point of view by allowing a direct measurement of an intra membrane concentration. The influence of

  9. Continuous Hydrolysis and Liquid–Liquid Phase Separation of an Active Pharmaceutical Ingredient Intermediate Using a Miniscale Hydrophobic Membrane Separator

    DEFF Research Database (Denmark)

    Cervera Padrell, Albert Emili; Morthensen, Sofie Thage; Lewandowski, Daniel Jacob

    2012-01-01

    Continuous hydrolysis of an active pharmaceutical ingredient intermediate, and subsequent liquid–liquid (L-L) separation of the resulting organic and aqueous phases, have been achieved using a simple PTFE tube reactor connected to a miniscale hydrophobic membrane separator. An alkoxide product...... a PTFE membrane with 28 cm2 of active area. A less challenging separation of water and toluene was achieved at total flow rates as high as 80 mL/min, with potential to achieve even higher flow rates. The operability and flexibility of the membrane separator and a plate coalescer were compared...

  10. Influence of phospholipid environment on the phosphatidylethanolamine: ceramide-phosphorylethanolamine transferase activity in rat liver plasma membranes.

    Science.gov (United States)

    Nikolova, M N; Petkova, D H; Koumanov, K S

    1992-03-01

    1. Plasma membranes were treated with phospholipase A2, phospholipase C or phospholipase D. The phosphatidylethanolamine:ceramide-phosphorylethanolamine transferase was deactivated by phospholipase C treatment, whereas phospholipase A2 and phospholipase D did not affect the enzyme. 2. Incorporation of phosphatidylethanolamine and phosphatidylglycerol into partially delipidated plasma membranes resulted in significant stimulation of the transferase, whereas inclusion of sphingomyelin and phosphatidylserine suppressed the enzyme activity. Our results suggest that phosphatidylserine is a regulator of sphingomyelin level in membranes. 3. The activity of phosphatidylethanolamine:ceramide-phosphorylethanolamine transferase was not influenced by the fluidity of its lipid environment.

  11. Effect of machining on the deformability of steel in surface-active medium at lower temperatures

    International Nuclear Information System (INIS)

    Gusti, E.Ya.; Babej, Yu.I.

    1977-01-01

    The effect of some machining methods of carbon steel, chromium steel, and chromium nickel steel, and that of low temperatures on the principle characteristics of formability during impact bending in air and a surface-active environment have been studied. The temperature decrease from the ambient to -80 deg is shown to reduce steel formability as evaluated by deflection (f) and to increase the forming force. The variation of these characteristics with lowering temperature, however, is greatly affected by machining process conditions. The FRHT (Friction-Hardening Treatment) on the white layer assures minimum ductility losses, and increases steel strength at low temperatures both in air and in the surface-active environment

  12. Kinetics and activity distribution of urease coencapsulated with hemoglobin within polyamide membranes.

    Science.gov (United States)

    Monshipouri, M; Neufeld, R J

    1992-01-01

    A 91.5% mass yield of urease and hemoglobin (Hb), co-encapsulated within polyamide membranes, was determined spectrophotometrically. The specific activity yield of microencapsulation was 84%, twofold higher than values previously reported, as a result of optimization of encapsulation conditions. The kinetic parameters and pH activity profiles of intracapsular urease were determined to be similar to those corresponding to the free enzyme. Similar activities were also observed for intact and microcapsule homogenate, indicating minimal mass transfer and diffusional limitation. The active configuration of the enzyme appears to remain intact upon microencapsulation. The application of a kinetic model for encapsulated urease further indicated that the kinetics were reaction-controlled with minimal mass transfer restrictions.

  13. Carbonic anhydrase activity of integral-functional complexes of thylakoid membranes of spinach chloroplasts

    Directory of Open Access Journals (Sweden)

    A. V. Semenihin

    2015-06-01

    Full Text Available Isolated thylakoid membranes were disrupted by treatment with nonionic detergents digitonin or dodecyl maltoside. Solubilized polypeptide complexes were separated by native gel charge shift electrophoresis. The position of ATP-synthase complex and its isolated catalytic part (CF1 within gel was determined using the color reaction for ATPase activity. Due to the presence of cytochromes, the red band in unstained gels corresponded to the cytochrome b6f complex. Localization of the cytochrome b6f complex, ATP synthase and coupling CF1 in the native gel was confirmed by their subunit composition determined after SDS-electrophoretic analysis. Carbonic anhydrase (CA activity in polypeptide zones of PS II, cytochrome b6f complex, and ATP-synthase CF1 was identified in native gels using indicator bromothymol blue. CA activity of isolated CF1 in solution was determined by infrared gas analysis as the rate of bicarbonate dehydration. The water-soluble acetazolamide, an inhibitor of CA, unlike lipophilic ethoxyzolamide inhibited CA activity of CF1. Thus, it was shown for the first time that ATP-synthase has a component which is capable of catalyzing the interconversion of forms of carbonic acid associated with proton exchange. The data obtained suggest the presence of multiple forms of carbonic anhydrase in the thylakoid membranes of spinach chloroplasts and confirm their involvement in the proton transfer to the ATP synthase.

  14. Fabrication Process Development for Light Deformable Mirrors

    Data.gov (United States)

    National Aeronautics and Space Administration — The project objective is to develop robust, reproductibble fabrication processes to realize functional deformable membrane mirrors (DM) for a space mission in which...

  15. Thermally-activated deformation in dispersion-hardened polycrystalline iron at room temperature

    DEFF Research Database (Denmark)

    Singh, Bachu Narain; Cotteril, P.

    1970-01-01

    The activation volume and dislocation velocity exponent have been obtained for polycrystalline iron in the extruded, extruded and annealed, and cold-rolled and annealed condition containing various amounts of alumina or zirconia particles, using the strain rate-change technique. It is found...

  16. Integrated carboxylic carbon nanotube pathways with membranes for voltage-activated humidity detection and microclimate regulation.

    Science.gov (United States)

    Pingitore, V; Miriello, D; Drioli, E; Gugliuzza, A

    2015-06-14

    This work describes some single walled carboxylic carbon nanotubes with outstanding transport properties when assembled in a 3D microarray working like a humidity membrane-sensor and an adjustable moisture regulator. Combined nano-assembly approaches are used to build up a better quality pathway through which assisted-charge and mass transport synchronically takes place. The structure-electrical response relationship is found, while controllable and tunable donor-acceptor interactions established at material interfaces are regarded as key factors for the accomplishment of charge transportation, enhanced electrical responses and adjustable moisture exchange. Raman and infrared spectroscopy provides indications about the fine structural and chemical features of the hybrid-composite membranes, resulting in perfect agreement with related morphology and electrical properties. Enhanced and modular electrical response to changes in the surrounding atmosphere is concerned with doping events, while assisted moisture regulation is discussed in relation to swelling and hopping actions. The electro-activated hybrid-composite membrane proposed in this work can be regarded as an attractive 'sense-to-act' precursor for smart long-distance monitoring systems with capability to adapt itself and provide local comfortable microenvironments.

  17. Energetics and molecular biology of active transport in bacterial membrane vesicles.

    Science.gov (United States)

    Kaback, H R; Ramos, S; Robertson, D E; Stroobant, P; Tokuda, H

    1977-01-01

    Bacterial membrane vesicles retain the same sidedness as the membrane in the intact cell and catalyze active transport of many solutes by a respiration-dependent mechanism that does not involve the generation of utilization of ATP or other high-energy phosphate compounds. In E. coli vesicles, most of these transport systems are coupled to an electrochemical gradient of protons (deltamuH+, interior negative and alkaline) generated primarily by the oxidation of D-lactate or reduced phenazine methosulfate via a membrane-bound respiratory chain. Oxygen or, under appropriate conditions, fumarate or nitrate can function as terminal electron acceptors, and the site at which deltamuH+ is generated is located before cytochrome b1 in the respiratory chain. Certain (N-dansyl)aminoalkyl-beta-D-galactopyranosides (Dns-gal) and N(2-nitro-4-azidophenyl)aminoalkyl 1-thio-beta-D-galactopyranosides (APG) are competitive inhibitors of lactose transport but are not transported themselves. Various fluorescence techniques, direct binding assays, and photoinactivation studies demonstrate that the great bulk of the lac carrier protein (ca. 95%) does not bind ligand in the absence of energy-coupling. Upon generation of a deltamuH+ (interior negative and alkaline), binding of Dns-gal and APG-dependent photoinactivation are observed. The data indicate that energy is coupled to the initial step in the transport process, and suggest that the lac carrier protein may be negatively charged.

  18. A packed bed membrane reactor for production of biodiesel using activated carbon supported catalyst.

    Science.gov (United States)

    Baroutian, Saeid; Aroua, Mohamed K; Raman, Abdul Aziz A; Sulaiman, Nik M N

    2011-01-01

    In this study, a novel continuous reactor has been developed to produce high quality methyl esters (biodiesel) from palm oil. A microporous TiO2/Al2O3 membrane was packed with potassium hydroxide catalyst supported on palm shell activated carbon. The central composite design (CCD) of response surface methodology (RSM) was employed to investigate the effects of reaction temperature, catalyst amount and cross flow circulation velocity on the production of biodiesel in the packed bed membrane reactor. The highest conversion of palm oil to biodiesel in the reactor was obtained at 70 °C employing 157.04 g catalyst per unit volume of the reactor and 0.21 cm/s cross flow circulation velocity. The physical and chemical properties of the produced biodiesel were determined and compared with the standard specifications. High quality palm oil biodiesel was produced by combination of heterogeneous alkali transesterification and separation processes in the packed bed membrane reactor. Copyright © 2010 Elsevier Ltd. All rights reserved.

  19. Advanced Supported Liquid Membranes for CO2 Control in Extravehicular Activity Applications

    Science.gov (United States)

    Wickham, David T.; Gleason, Kevin J.; Engel, Jeffrey R.; Cowley, Scott W.; Chullen, Cinda

    2014-01-01

    Developing a new, robust, portable life support system (PLSS) is currently a high priority for NASA in order to support longer and safer extravehicular activity (EVA) missions. One of the critical PLSS functions is maintaining the carbon dioxide (CO2) concentration in the suit at acceptable levels. Although the Metal Oxide (MetOx) canister has worked well, it has a finite CO2 adsorption capacity. Consequently, the unit would have to be larger and heavier to extend EVA times. Therefore, new CO2 control technologies must be developed to meet mission objectives without increasing the size of the PLSS. Although recent work has centered on sorbents that can be regenerated during the EVA, this strategy increases the system complexity and power consumption. A simpler approach is to use a membrane that selectively vents CO2 to space. A membrane has many advantages over current technology: it is a continuous system with no theoretical capacity limit, it requires no consumables, and it requires no hardware for switching beds between absorption and regeneration. Unfortunately, conventional gas separation membranes do not have adequate selectivity for use in the PLSS. However, the required performance could be obtained with a supported liquid membrane (SLM), which consists of a micro porous material filled with a liquid that selectively reacts with CO2 over oxygen (O2). In a current Phase II SBIR project, Reaction Systems has developed a new reactive liquid, which has effectively zero vapor pressure making it an ideal candidate for use in an SLM. The SLM function has been demonstrated with representative pressures of CO2, O2, and water (H2O). In addition to being effective for CO2 control, the SLM also vents moisture to space. Therefore, this project has demonstrated the feasibility of using an SLM to control CO2 in an EVA application.

  20. Membrane Active Small Molecules Show Selective Broad Spectrum Antibacterial Activity with No Detectable Resistance and Eradicate Biofilms.

    Science.gov (United States)

    Hoque, Jiaul; Konai, Mohini M; Gonuguntla, Spandhana; Manjunath, Goutham B; Samaddar, Sandip; Yarlagadda, Venkateswarlu; Haldar, Jayanta

    2015-07-23

    Treating bacterial biofilms with conventional antibiotics is limited due to ineffectiveness of the drugs and higher propensity to develop bacterial resistance. Development of new classes of antibacterial therapeutics with alternative mechanisms of action has become imperative. Herein, we report the design, synthesis, and biological evaluations of novel membrane-active small molecules featuring two positive charges, four nonpeptidic amide groups, and variable hydrophobic/hydrophilic (amphiphilic) character. The biocides synthesized via a facile methodology not only displayed good antibacterial activity against wild-type bacteria but also showed high activity against various drug-resistant bacteria such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecium (VRE), and β-lactam-resistant Klebsiella pneumoniae. Further, these biocides not only inhibited the formation of biofilms but also disrupted the established S. aureus and E. coli biofilms. The membrane-active biocides hindered the propensity to develop bacterial resistance. Moreover, the biocides showed negligible toxicity against mammalian cells and thus bear potential to be used as therapeutic agents.

  1. A novel membrane sensor controls the localization and ArfGEF activity of bacterial RalF.

    Directory of Open Access Journals (Sweden)

    Marcia Folly-Klan

    Full Text Available The intracellular bacterial pathogen Legionella pneumophila (Lp evades destruction in macrophages by camouflaging in a specialized organelle, the Legionella-containing vacuole (LCV, where it replicates. The LCV maturates by incorporating ER vesicles, which are diverted by effectors that Lp injects to take control of host cell membrane transport processes. One of these effectors, RalF, recruits the trafficking small GTPase Arf1 to the LCV. LpRalF has a Sec7 domain related to host ArfGEFs, followed by a capping domain that intimately associates with the Sec7 domain to inhibit GEF activity. How RalF is activated to function as a LCV-specific ArfGEF is unknown. We combined the reconstitution of Arf activation on artificial membranes with cellular expression and Lp infection assays, to analyze how auto-inhibition is relieved for LpRalF to function in vivo. We find that membranes activate LpRalF by about 1000 fold, and identify the membrane-binding region as the region that inhibits the Sec7 active site. It is enriched in aromatic and positively charged residues, which establish a membrane sensor to control the GEF activity in accordance with specific lipid environments. A similar mechanism of activation is found in RalF from Rickettsia prowazekii (Rp, with a different aromatic/charged residues ratio that results in divergent membrane preferences. The membrane sensor is the primary determinant of the localization of LpRalF on the LCV, and drives the timing of Arf activation during infection. Finally, we identify a conserved motif in the capping domain, remote from the membrane sensor, which is critical for RalF activity presumably by organizing its active conformation. These data demonstrate that RalF proteins are regulated by a membrane sensor that functions as a binary switch to derepress ArfGEF activity when RalF encounters a favorable lipid environment, thus establishing a regulatory paradigm to ensure that Arf GTPases are efficiently activated at

  2. Electrocatalysts and their Supporting Materials for Proton Exchange Membrane Fuel Cells: Activity and Durability Studies

    DEFF Research Database (Denmark)

    Permyakova, Anastasia Aleksandrovna

    other methods. The thesis begins with an introduction in Chapter 1 providing an overview of fuel cells, their associated reaction mechanisms, catalysts and catalysts supports. Chapter 2 presents the theoretical background to the study including equipment and the techniques used to analyse the catalysts......This thesis describes investigations conducted exploring the activity, stability and durability of supported nano-particulate, bulk and thin film electrocatalysts used in proton exchange membrane fuel cells (PEMFCs). The effects of different factors and conditions on the reactions involved...

  3. Spermine modulates fungal morphogenesis and activates plasma membrane H+-ATPase during yeast to hyphae transition

    Directory of Open Access Journals (Sweden)

    Antônio Jesus Dorighetto Cogo

    2018-02-01

    Full Text Available Polyamines play a regulatory role in eukaryotic cell growth and morphogenesis. Despite many molecular advances, the underlying mechanism of action remains unclear. Here, we investigate a mechanism by which spermine affects the morphogenesis of a dimorphic fungal model of emerging relevance in plant interactions, Yarrowia lipolytica, through the recruitment of a phytohormone-like pathway involving activation of the plasma membrane P-type H+-ATPase. Morphological transition was followed microscopically, and the H+-ATPase activity was analyzed in isolated membrane vesicles. Proton flux and acidification were directly probed at living cell surfaces by a non-invasive selective ion electrode technique. Spermine and indol-3-acetic acid (IAA induced the yeast-hypha transition, influencing the colony architecture. Spermine induced H+-ATPase activity and H+ efflux in living cells correlating with yeast-hypha dynamics. Pharmacological inhibition of spermine and IAA pathways prevented the physio-morphological responses, and indicated that spermine could act upstream of the IAA pathway. This study provides the first compelling evidence on the fungal morphogenesis and colony development as modulated by a spermine-induced acid growth mechanism analogous to that previously postulated for the multicellular growth regulation of plants.

  4. Antifungal activity of alkanols against Zygosaccharomyces bailii and their effects on fungal plasma membrane.

    Science.gov (United States)

    Fujita, Ken-Ichi; Fujita, Tomoko; Kubo, Isao

    2008-10-01

    A series of aliphatic primary alkanols from C(6) to C(13) were tested for antifungal activity against a food spoilage fungus Zygosaccharomyces bailii using a broth dilution method and were compared for their effects against Saccharomyces cerevisiae and Z. rouxii. Decanol (C(10)) was found to be the most potent fungicide against Z. bailii at a minimum fungicidal concentration of 50 microg/ml (0.31 mM), whereas undecanol (C(11)) was found to be the most potent fungistatic at a minimum inhibitory concentration of 25 microg/ml (0.14 mM). The time-kill curve study showed that decanol was fungicidal against Z. bailii at any growth stage. Octanol (C(8)) increased plasma membrane fluidity in the spheroplast cells of S. cerevisiae. The primary antifungal action of alkanols comes from their ability to disrupt the native membrane-associated function of integral proteins nonspecifically as nonionic surface-active agents (surfactants). The antifungal activity of decanol against Z. bailii was slightly enhanced in combination with anethole. (c) 2008 John Wiley & Sons, Ltd.

  5. Gastrointestinal Hormone Cholecystokinin Increases P-Glycoprotein Membrane Localization and Transport Activity in Caco-2 Cells.

    Science.gov (United States)

    Yano, Kentaro; Shimizu, Saori; Tomono, Takumi; Ogihara, Takuo

    2017-09-01

    It was reported that stimulation of taste receptor type 2 member 38 by a bitter substance, phenylthiocarbamide (PTC), increased P-glycoprotein (P-gp) mRNA level and transport activity via release of the gastrointestinal hormone cholecystokinin-8 (CCK-8) at 9 h. Therefore, we hypothesized that CCK-8 and PTC might also regulate P-gp activity more rapidly via a different mechanism. As a result, we found that the pretreatment of human colon adenocarcinoma (Caco-2) cells with 10-mM PTC significantly decreased the intracellular accumulation of P-gp substrate rhodamine 123 (Rho123) compared with the control after 90-min incubation. Moreover, CCK-8 treatments significantly reduced the accumulation of Rho123 within 30 min, compared with the control. On the other hand, when Caco-2 cells were pretreated with PTC, the efflux ratio of Rho123 was significantly increased compared with control. The efflux ratio of Rho123 in CCK-8 treatment cells was also significantly increased compared with control. Furthermore, CCK-8 increased the phosphorylation of the scaffold proteins ezrin, radixin, and moesin, which regulate translocation of P-gp to the plasma membrane. Therefore, our results indicate that PTC induced release of CCK-8, which in turn induced the phosphorylation of ezrin, radixin, and moesin proteins, leading to upregulation of P-gp transport activity via increased membrane localization of P-gp. Copyright © 2017 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

  6. Layer-by-layer structured polysaccharides-based multilayers on cellulose acetate membrane: Towards better hemocompatibility, antibacterial and antioxidant activities

    Science.gov (United States)

    Peng, Lincai; Li, Hui; Meng, Yahong

    2017-04-01

    The development of multifunctional cellulose acetate (CA) membranes with enhanced hemocompatibility and antibacterial and antioxidant activities is extremely important for biomedical applications. In this work, significant improvements in hemocompatibility and antibacterial and antioxidant activities of cellulose acetate (CA) membranes were achieved via layer-by-layer (LBL) deposition of chitosan (CS) and water-soluble heparin-mimicking polysaccharides (i.e., sulfated Cantharellus cibarius polysaccharides, SCP) onto their surface. The surface chemical compositions, growth manner, surface morphologies, and wetting ability of CS/SCP multilayer-modified CA membranes were characterized, respectively. The systematical evaluation of hemocompatibility revealed that CS/SCP multilayer-modified CA membranes significantly improved blood compatibility including resistance to non-specific protein adsorption, suppression of platelet adhesion and activation, prolongation of coagulation times, inhibition of complement activation, as well as reduction in blood hemolysis. Meanwhile, CS/SCP multilayer-modified CA membranes exhibited strong growth inhibition against Escherichia coli and Staphylococcus aureus, as well as high scavenging abilities against superoxide and hydroxyl radicals. In summary, the CS/SCP multilayers could confer CA membranes with integrated hemocompatibility and antibacterial and antioxidant activities, which might have great potential application in the biomedical field.

  7. Mitochondrial ATPase activity and membrane fluidity changes in rat liver in response to intoxication with Buckthorn (Karwinskia humboldtiana

    Directory of Open Access Journals (Sweden)

    Margarita Cid-Hernández

    2015-01-01

    Full Text Available BACKGROUND: Karwinskia humboldtiana (Kh is a poisonous plant of the rhamnacea family. To elucidate some of the subcellular effects of Kh toxicity, membrane fluidity and ATPase activities as hydrolytic and as proton-pumping activity were assessed in rat liver submitochondrial particles. Rats were randomly assigned into control non-treated group and groups that received 1,1.5 and 2 g/Kg body weight of dry powder of Kh fruit, respectively. Rats were euthanized at day 1 and 7 after treatment RESULTS: Rats under Kh treatment at all dose levels tested, does not developed any neurologic symptoms. However, we detected alterations in membrane fluidity and ATPase activity. Lower dose of Kh on day 1 after treatment induced higher mitochondrial membrane fluidity than control group. This change was strongly correlated with increased ATPase activity and pH gradient driven by ATP hydrolysis. On the other hand, membrane fluidity was hardly affected on day 7 after treatment with Kh. Surprisingly, the pH gradient driven by ATPase activity was significantly higher than controls despite an diminution of the hydrolytic activity of ATPase CONCLUSIONS: The changes in ATPase activity and pH gradient driven by ATPase activity suggest an adaptive condition whereby the fluidity of the membrane is altered

  8. Active faults and related Late Quaternary deformation along the Northwestern Himalayan Frontal Zone, India

    Directory of Open Access Journals (Sweden)

    T. Nakata

    2003-06-01

    Full Text Available Numerous newly-identified traces of active faults in the Himalayan foothill zone along the HFF around Chandigarh, in Pinjore Dun, along the piedmont zone of the Lower Siwalik hill front and within the Lower Tertiary hill range reveal the pattern of thrust and strike-slip faulting, striking parallel to the principal structural trend (NNW-SSE of the orogenic belt. The active Chandigarh Fault, Pinjore Garden Fault and Barsar thrust have vertically dislocated, warped and backtilted fluvial and alluvial-fan surfaces made up of Late Pleistocene-Holocene sediments. West- and southwest-facing fault scarplets with heights ranging from 12 to 50 m along these faults suggest continued tectonic movement through Late Pleistocene to recent times. Gentle warping and backtilting of the terraces on the hanging wall sides of the faults indicate fault-bend folding. These active faults are the manifestation of north-dipping imbricated thrust faults branching out from the major fault systems like the Main Boundary Fault (MBF and Himalayan Frontal Fault (HFF, probably merging down northward into a décollement. The Taksal Fault, striking NNW-SSE, shows prominent right-lateral movement marked by lateral offset of streams and younger Quaternary terraces and occupies a narrow deep linear valley along the fault trace. Right stepping along this fault has resulted in formation of a small pull-apart basin. Fault scarplets facing ENE and WSW are the manifestation of dip-slip movement. This fault is an example of slip-partitioning between the strike-slip and thrust faults, suggesting ongoing oblique convergence of the Indian plate and northward migration of a tectonic sliver. Slip rate along the Taksal Fault has been calculated as 2.8 mm/yr. Preliminary trench investigation at the base of the Chandigarh Fault Scarp has revealed total displacement of 3.5 m along a low angle thrust fault with variable dip of 20° to 46° due northeast, possibly the result of one

  9. Biological Activity Alterations of Human Amniotic Membrane Pre and Post Irradiation Tissue Banking.

    Science.gov (United States)

    Nemr, Waleed; Bashandy, A S; Araby, Eman; Khamiss, O

    Innate immunity of Human Amniotic Membrane (HAM) and its highly active secretome that rich with various types of growth factors and anti-inflammatory substances proposed it as a promising material for many medical studies and applications. This study evaluate the biological activity of cultivated HAM pre and post tissue banking process in which freeze-dried HAM was sterilized by 25 KGray (kGy) dose of γ radiation. The HAM's antimicrobial activity, viability, growth of isolated human amniotic epithelial cells (HAECs), hematopoietic stimulation of co-cultivated murine bone marrow cells (mammalian model), scaffold efficiency for fish brain building up (non-mammalian model) and self re-epithelialization after trypsin denuding treatment were examined as supposed biological activity features. Native HAM revealed viability indications and was active to kill all tested microorganisms; 6 bacterial species (3 Gram-positive and 3 Gram-negative) and Candida albicans as a pathogenic fungus. Also, HAM activity promoted colony formation of murine hematopoietic cells, Tilapia nilotica brain fragment building-up and self re-epithelialization after trypsin treatment. In contrary, radiation-based tissue banking of HAM caused HAM cellular death and consequently lacked almost all of examined biological activity features. Viable HAM was featured with biological activity than fixed HAM prepared by irradiation tissue banking.

  10. Stabilization of axisymmetric modes and the effects of plasma deformability in elongated tokamak plasmas using active feedback coils inside and outside the vacuum vessel

    International Nuclear Information System (INIS)

    Ward, D.J.; Hofmann, F.

    1992-01-01

    In this paper we study the feedback stabilization of the vertical instability in a highly elongated (κ = 3) TCV plasma using a combination of active coils inside and outside the vacuum vessel. The TCV is a tokamak under construction at the CRPP in Lausanne. It is designed to create plasmas with a variety of cross-sectional shapes with elongations up to κ = 3. We calculate axisymmetric stability using the NOVA-W code. It has been observed that the deformation of the eigenfunction plays a crucial role in the ability of the feedback system to successfully stabilize the plasma. It is important to understand the deformation of the eigenfunction resulting from a particular choice of active coils in order to choose an appropriate set of magnetic measurements to determine the vertical displacement. The use of internal and external active coils imposes a particular deformation that allows one to choose a small number of well placed measurements. A large number of magnetic measurements distributed around the plasma does not necessarily improve matters, and in fact can be much less effective than a smaller number of well placed measurements. The deformation of the eigenfunction is also strongly affected by the distribution of currents in the external coils. (author) 3 figs., 3 refs

  11. High-protein-PUFA supplementation, red blood cell membranes, and plasma antioxidant activity in volleyball athletes.

    Science.gov (United States)

    Malaguti, Marco; Baldini, Marta; Angeloni, Cristina; Biagi, Pierluigi; Hrelia, Silvana

    2008-06-01

    The authors evaluated the role of a high-protein, low-calorie, polyunsaturated fatty-acid (PUFA) -supplemented diet on anthropometric parameters, erythrocyte-membrane fatty-acid composition, and plasma antioxidant defenses of nonprofessional volleyball athletes. The athletes were divided in two groups: One (n = 5) followed the Mediterranean diet, and the other (n = 6) followed a high-protein, low-calorie diet with a 3-g/day fish-oil supplementation. All the athletes had anthropometric measurements taken, both at the beginning and at the end of the study, which lasted for 2 months. Body-mass index and total body fat were significantly diminished in the second group, while they remained unchanged in the first. Plasma total antioxidant activity (TAA) was significantly increased in the plasma of both groups, with no differences between the groups, suggesting that physical activity, not the different diets, is the main contributor to the increase of plasma TAA. The second group showed a significant increase in erythrocyte-membrane PUFA content and in the unsaturation index value (UI) because of the fish-oil supplementation.A high-protein, low-carbohydrate, fish-oil-supplemented diet seems to be useful only when the aim of the diet is to obtain weight loss in a short-term period. The significant increase in the UI of erythrocyte membranes indicates the potential for harm, because a high intake of PUFA might increase susceptibility to lipid peroxidation not counterbalanced by a higher increase in TAA. Adherence to the Mediterranean diet seems to be the better choice.

  12. Effects of deuterated water upon specific activity of some marker enzymes for cytosol and plasmatic membrane

    International Nuclear Information System (INIS)

    Buzgariu, Wanda; Coroiu, Viorica; Moldovan, Lucia; Titescu, G.; Stefanescu, I.

    2004-01-01

    Recently, numerous studies were devoted to the effects of an increased environmental deuterium concentration on physiological characteristics of various biological systems, from monocellular organisms up to mammals. Within these preoccupations the experiments on enzyme activity and parameters are of special interest since they throw light upon the mechanisms in metabolic biochemical reactions (glycolysis, photosynthesis, transport across membranes, etc). The present work concerns the effects of heavy water upon the activity of some enzymes (dehydrogenase-LDH lactate and 5' nucleotidase) implied in different metabolic pathways, serving as functional indicators for some cellular compartments such as the cytosols and cellular membranes. Enzyme activity was determined by growing for 6 days the cells (Hep 2, CHO, fibroblasts) in deuterated culture media at different concentration levels (20%, 40%, 65% si 90%), as well as in a reaction medium deuterated at 99.96%. In case of the first experimental run the LDH activity was monitored for the three cellular lines (Hep 2, CHO, fibroblasts) for different time intervals (1 d, 3 d and 6 d). After the first 24 h of cells' exposure the activity values were similar regardless of the heavy water concentration in the medium. Exposing the cells for longer time (6 days) led to modifications of LDH activity. In contrast to the case of media with relatively moderate D 2 O content, cell growing in conditions of intense deuteration 65% and 90 % D 2 O) led to an increase of cytosolic enzyme activity of about 50%. In case of 5' nucleotidase after 6 days of cell cultivation in deuteration conditions the activity decreased to 50% and 70% from the value corresponding to normal conditions for cell growth. This diminution of the activity was characteristic for the media with 65% and 90% D 2 O. In the second experimental run the activities of dehydrogenase lactate and 5' nucleotidase from the cellular homogenate obtained from cells grown in

  13. Modulation of the interface between polyester and spent coffee grounds in polysaccharide membranes: Preparation, cell proliferation, antioxidant activity and tyrosinase activity.

    Science.gov (United States)

    Wu, Chin-San

    2017-09-01

    The structural, antioxidant and cytocompatibility properties of membranes prepared from polyhydroxyalkanoate (PHA) and spent coffee ground (SCG) blends (PHA/SCG) were studied. Acrylic acid-grafted PHA (PHA-g-AA) was used to enhance the desirable characteristics of these membranes, which had better tensile properties than the corresponding PHA/SCG membranes. The water resistance of the PHA-g-AA/SCG membranes was greater than that of the PHA/SCG membranes, and a cytocompatibility evaluation with mouse normal tail fibroblasts (FBs) indicated that both materials were nontoxic. Cell cycle assays of FBs on PHA/SCG and PHA-g-AA/SCG membrane samples were not affected by the DNA content related to damage. Moreover, SCG enhanced the saccharide and polyphenol contents, and antioxidant properties, of the PHA-g-AA/SCG and PHA/SCG membranes. Therefore, we analysed the effects of these compounds' membranes on melanogenesis in B16-F10 melanoma cells. The results demonstrated that PHA/SCG and PHA-g-AA/SCG membranes reduced cellular tyrosinase activities in vitro. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Use of stable emulsion to improve stability, activity, and enantioselectivity of lipase immobilized in a membrane reactor.

    Science.gov (United States)

    Giorno, L; Li, N; Drioli, E

    2003-12-20

    The enantiocatalytic performance of immobilized lipase in an emulsion membrane reactor using stable emulsion prepared by membrane emulsification technology was studied. The production of optical pure (S)-naproxen from racemic naproxen methyl ester was used as a model reaction system. The O/W emulsion, containing the substrate in the organic phase, was fed to the enzyme membrane reactor from shell-to-lumen. The enzyme was immobilized in the sponge layer (shell side) of capillary polyamide membrane with 50 kDa cut-off. The aqueous phase was able to permeate through the membrane while the microemulsion was retained by the thin selective layer. Therefore, the substrate was kept in the enzyme-loaded membrane while the water-soluble product was continuously removed from the reaction site. The results show that lipase maintained stable activity during the entire operation time (more than 250 h), showing an enantiomeric excess (96 +/- 2%) comparable to the free enzyme (98 +/- 1%) and much higher compared to similar lipase-loaded membrane reactors used in two-separate phase systems (90%). The results demonstrate that immobilized enzymes can achieve high stability as well as high catalytic activity and enantioselectivity. Copyright 2003 Wiley Periodicals, Inc.

  15. Surface Functionalization of Polyethersulfone Membrane with Quaternary Ammonium Salts for Contact-Active Antibacterial and Anti-Biofouling Properties

    Directory of Open Access Journals (Sweden)

    Xiao Hu

    2016-05-01

    Full Text Available Biofilm is a significant cause for membrane fouling. Antibacterial-coated surfaces can inhibit biofilm formation by killing bacteria. In this study, polyethersulfone (PES microfiltration membrane was photografted by four antibiotic quaternary ammonium compounds (QACs separately, which were synthesized from dimethylaminoethyl methacrylate (DMAEMA by quaternization with butyl bromide (BB, octyl bromide (OB, dodecyl bromide (DB, or hexadecyl bromide (HB. XPS, ATR-FTIR, and SEM were used to confirm the surfaces’ composition and morphology. After modification, the pores on PES-g-DMAEMA-BB and PES-g-DMAEMA-OB were blocked, while PES-g-DMAEMA-DB and PES-g-DMAEMA-HB were retained. We supposed that DMAEMA-BB and DMAEMA-OB aggregated on the membrane surface due to the activities of intermolecular or intramolecular hydrogen bonds. Bacteria testing found the antibacterial activities of the membranes increased with the length of the substituted alkyl chain. Correspondingly, little bacteria were observed on PES-g-DMAEMA-DB and PES-g-DMAEMA-HB by SEM. The antifouling properties were investigated by filtration of a solution of Escherichia coli. Compared with the initial membrane, PES-g-DMAEMA-DB and PES-g-DMAEMA-HB showed excellent anti-biofouling performance with higher relative flux recovery (RFR of 88.3% and 92.7%, respectively. Thus, surface functionalization of the PES membrane with QACs can prevent bacteria adhesion and improve the anti-biofouling activity by the contact-active antibacterial property.

  16. Effect of yeast biomass with high content of carotenoids on erythrocyte deformability, NO production and Na,K-ATPase activity in healthy and LPS treated rats.

    Science.gov (United States)

    Radosinska, J; Mezesova, L; Okruhlicova, L; Frimmel, K; Breierova, E; Bartekova, M; Vrbjar, N

    2016-11-25

    Measurements of red blood cell (RBC) deformability together with estimation of NO-synthase activity and Na,K-ATPase activity were used for characterization of RBC functionality in rats subjected to single dose of Escherichia coli lipopolysaccharides (LPS) at a dose of 1 mg/kg. We hypothesized that LPS might initiate a malfunction of RBC. We also investigated the potential effect of carotenoids (10 mg/kg/day) produced in red yeast biomass of Rhodotorula glutinis on RBC in LPS-challenged rats. LPS significantly reduced the deformability of RBC (by 14%) together with decrease of NO-synthase activity by 20%. Daily supplementation of carotenoids for 10 days attenuated the LPS-induced injury, as observed by 22% increase of RBC deformability and 23% increase of NO-synthase activity. The activity of Na,K-ATPase was also improved probably due to increased number of active enzyme molecules as indicated by 66% enhancement of Vmax value, hence maintaining the activity of erythrocyte Na,K-ATPase to the level even higher as compared with healthy control animals. It may be concluded that administration of yeast biomass with high content of carotenoids resulted in advanced function of erythrocytes as concerns their ability to squeeze through narrow capillaries of the circulation, better intrinsic production of NO and improvement of intracellular homeostasis of sodium.

  17. Inhibition of membrane lipid-independent protein kinase Calpha activity by phorbol esters, diacylglycerols, and bryostatin-1.

    Science.gov (United States)

    Slater, S J; Taddeo, F J; Mazurek, A; Stagliano, B A; Milano, S K; Kelly, M B; Ho, C; Stubbs, C D

    1998-09-04

    The activity of membrane-associated protein kinase C (PKC) has previously been shown to be regulated by two discrete high and low affinity binding regions for diacylglycerols and phorbol esters (Slater, S. J., Ho, C., Kelly, M. B., Larkin, J. D., Taddeo, F. J., Yeager, M. D., and Stubbs, C. D. (1996) J. Biol. Chem. 271, 4627-4631). PKC is also known to interact with both cytoskeletal and nuclear proteins; however, less is known concerning the mode of activation of this non-membrane form of PKC. By using the fluorescent phorbol ester, sapintoxin D (SAPD), PKCalpha, alone, was found to possess both low and high affinity phorbol ester-binding sites, showing that interaction with these sites does not require association with the membrane. Importantly, a fusion protein containing the isolated C1A/C1B (C1) domain of PKCalpha also bound SAPD with low and high affinity, indicating that the sites may be confined to this domain rather than residing elsewhere on the enzyme molecule. Both high and low affinity interactions with native PKCalpha were enhanced by protamine sulfate, which activates the enzyme without requiring Ca2+ or membrane lipids. However, this "non-membrane" PKC activity was inhibited by the phorbol ester 4beta-12-O-tetradecanoylphorbol-13-acetate (TPA) and also by the fluorescent analog, SAPD, opposite to its effect on membrane-associated PKCalpha. Bryostatin-1 and the soluble diacylglycerol, 1-oleoyl-2-acetylglycerol, both potent activators of membrane-associated PKC, also competed for both low and high affinity SAPD binding and inhibited protamine sulfate-induced activity. Furthermore, the inactive phorbol ester analog 4alpha-TPA (4alpha-12-O-tetradecanoylphorbol-13-acetate) also inhibited non-membrane-associated PKC. In keeping with these observations, although TPA could displace high affinity SAPD binding from both forms of the enzyme, 4alpha-TPA was only effective at displacing high affinity SAPD binding from non-membrane-associated PKC. 4alpha-TPA also

  18. Latent nitrate reductase activity is associated with the plasma membrane of corn roots

    Science.gov (United States)

    Ward, M. R.; Grimes, H. D.; Huffaker, R. C.

    1989-01-01

    Latent nitrate reductase activity (NRA) was detected in corn (Zea mays L., Golden Jubilee) root microsome fractions. Microsome-associated NRA was stimulated up to 20-fold by Triton X-100 (octylphenoxy polyethoxyethanol) whereas soluble NRA was only increased up to 1.2-fold. Microsome-associated NRA represented up to 19% of the total root NRA. Analysis of microsomal fractions by aqueous two-phase partitioning showed that the membrane-associated NRA was localized in the second upper phase (U2). Analysis with marker enzymes indicated that the U2 fraction was plasma membrane (PM). The PM-associated NRA was not removed by washing vesicles with up to 1.0 M NACl but was solubilized from the PM with 0.05% Triton X-100. In contrast, vanadate-sensitive ATPase activity was not solubilized from the PM by treatment with 0.1% Triton X-100. The results show that a protein capable of reducing nitrate is embedded in the hydrophobic region of the PM of corn roots.

  19. Three-Dimensional Crustal Architecture Beneath the Sikkim Himalaya and Its Relationship to Active Deformation

    Science.gov (United States)

    Paul, Himangshu; Mitra, Supriyo

    2017-10-01

    We study the 3-D variation of the crustal structure of the Sikkim Himalaya using broadband seismological data acquired from a focused network of seven stations spanning the Lesser, Higher, and Tethyan Himalaya. Common conversion point stacking of receiver functions recorded along an across-strike profile of the Himalaya reveals first-order northward dip on the Main Himalayan Thrust (MHT), a midcrustal discontinuity and the Moho, along with higher-order lateral variations. Three-dimensional images generated from joint inversion of receiver functions and surface wave dispersions show that the MHT has a ramp-flat-ramp geometry. The ramps are located beneath the Lesser Himalaya and the Tethyan Himalaya with dips of ˜7∘ and ˜15∘, respectively, connected by flat segments. The ramp beneath the Lesser Himalaya forms a dome structure, upwarping the thrust sheets associated with the Peling and Main Central Thrust. The erosional surface of this dome forms the arcuate geometry of thrusts observed in the Lesser Himalaya. The thickness of the underthrust Indian crust is 35-42 km and has an average VS of 3.63 km/s, similar to that of the Indian Shield crust. The Moho also has dome-like structures separated by elongated, deeper sections trending NW-SE. These are intersected by steeply dipping transverse low-velocity zones, oblique to the strike of the Himalaya. We conjecture that these low-velocity zones are the dextral-strike slip faults known to be active beneath the Sikkim Himalaya. The observed alternate shallow and deep segments of the Moho must be a consequence of several cycles of strike-slip displacement on these transverse faults.

  20. Active tectonic deformation along rejuvenated faults in tropical Borneo: Inferences obtained from tectono-geomorphic evaluation

    Science.gov (United States)

    Mathew, Manoj Joseph; Menier, David; Siddiqui, Numair; Kumar, Shashi Gaurav; Authemayou, Christine

    2016-08-01

    active folding of the Rajang Group fold-thrust belt to present and these events reactivated old major faults and minor related dislocations. From geomorphic analysis associated with sedimentary record, we posit that the terrain could have undergone high uplift rates since 5 Ma or multi-phased uplift with periodic intermittent pulses of high and low uplift rates.

  1. Nonlinear Deformable-body Dynamics

    CERN Document Server

    Luo, Albert C J

    2010-01-01

    "Nonlinear Deformable-body Dynamics" mainly consists in a mathematical treatise of approximate theories for thin deformable bodies, including cables, beams, rods, webs, membranes, plates, and shells. The intent of the book is to stimulate more research in the area of nonlinear deformable-body dynamics not only because of the unsolved theoretical puzzles it presents but also because of its wide spectrum of applications. For instance, the theories for soft webs and rod-reinforced soft structures can be applied to biomechanics for DNA and living tissues, and the nonlinear theory of deformable bodies, based on the Kirchhoff assumptions, is a special case discussed. This book can serve as a reference work for researchers and a textbook for senior and postgraduate students in physics, mathematics, engineering and biophysics. Dr. Albert C.J. Luo is a Professor of Mechanical Engineering at Southern Illinois University, Edwardsville, IL, USA. Professor Luo is an internationally recognized scientist in the field of non...

  2. Lpx1p links glucose-induced calcium signaling and plasma membrane H+-ATPase activation in Saccharomyces cerevisiae cells.

    Science.gov (United States)

    Castanheira, Diogo Dias; Santana, Eduardo Perovano; Godoy-Santos, Fernanda; Diniz, Raphael Hermano Santos; Faria-Oliveira, Fábio; Pereira, Renata Rebeca; Trópia, Maria José Magalhães; Castro, Ieso Miranda; Brandão, Rogelio Lopes

    2018-02-01

    In yeast, as in other eukaryotes, calcium plays an essential role in signaling transduction to regulate different processes. Many pieces of evidence suggest that glucose-induced activation of plasma membrane H+-ATPase, essential for yeast physiology, is related to calcium signaling. Until now, no protein that could be regulated by calcium in this context has been identified. Lpx1p, a serine-protease that is also involved in the glucose-induced activation of the plasma membrane H+-ATPase, could be a candidate to respond to intracellular calcium signaling involved in this process. In this work, by using different approaches, we obtained many pieces of evidence suggesting that the requirement of calcium signaling for activation of the plasma membrane H+-ATPase is due to its requirement for activation of Lpx1p. According to the current model, activation of Lpx1p would cause hydrolysis of an acetylated tubulin that maintains the plasma membrane H+-ATPase in an inactive state. Therefore, after its activation, Lpx1p would hydrolyze the acetylated tubulin making the plasma membrane H+-ATPase accessible for phosphorylation by at least one protein kinase. © FEMS 2017. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  3. Unique battery with an active membrane separator having uniform physico-chemically functionalized ion channels and a method making the same

    Science.gov (United States)

    Gerald, II, Rex E.; Ruscic, Katarina J [Chicago, IL; Sears, Devin N [Spruce Grove, CA; Smith, Luis J [Natick, MA; Klingler, Robert J [Glenview, IL; Rathke, Jerome W [Homer Glen, IL

    2012-02-21

    The invention relates to a unique battery having an active, porous membrane and method of making the same. More specifically the invention relates to a sealed battery system having a porous, metal oxide membrane with uniform, physicochemically functionalized ion channels capable of adjustable ionic interaction. The physicochemically-active porous membrane purports dual functions: an electronic insulator (separator) and a unidirectional ion-transporter (electrolyte). The electrochemical cell membrane is activated for the transport of ions by contiguous ion coordination sites on the interior two-dimensional surfaces of the trans-membrane unidirectional pores. The membrane material is designed to have physicochemical interaction with ions. Control of the extent of the interactions between the ions and the interior pore walls of the membrane and other materials, chemicals, or structures contained within the pores provides adjustability of the ionic conductivity of the membrane.

  4. Sch proteins are localized on endoplasmic reticulum membranes and are redistributed after tyrosine kinase receptor activation

    DEFF Research Database (Denmark)

    Lotti, L V; Lanfrancone, L; Migliaccio, E

    1996-01-01

    The intracellular localization of Shc proteins was analyzed by immunofluorescence and immunoelectron microscopy in normal cells and cells expressing the epidermal growth factor receptor or the EGFR/erbB2 chimera. In unstimulated cells, the immunolabeling was localized in the central perinuclear...... and endocytic structures, such as coated pits and endosomes, and with the peripheral cytosol. Receptor activation in cells expressing phosphorylation-defective mutants of Shc and erbB-2 kinase showed that receptor autophosphorylation, but not Shc phosphorylation, is required for redistribution of Shc proteins....... The rough endoplasmic reticulum localization of Shc proteins in unstimulated cells and their massive recruitment to the plasma membrane, endocytic structures, and peripheral cytosol following receptor tyrosine kinase activation could account for multiple putative functions of the adaptor protein....

  5. Outer Membrane Protein 25 of Brucella Activates Mitogen-Activated Protein Kinase Signal Pathway in Human Trophoblast Cells

    Directory of Open Access Journals (Sweden)

    Jing Zhang

    2017-12-01

    Full Text Available Outer membrane protein 25 (OMP25, a virulence factor from Brucella, plays an important role in maintaining the structural stability of Brucella. Mitogen-activated protein kinase (MAPK signal pathway widely exists in eukaryotic cells. In this study, human trophoblast cell line HPT-8 and BALB/c mice were infected with Brucella abortus 2308 strain (S2308 and 2308ΔOmp25 mutant strain. The expression of cytokines and activation of MAPK signal pathway were detected. We found that the expressions of tumor necrosis factor-α, interleukin-1, and interleukin-10 (IL-10 were increased in HPT-8 cells infected with S2308 and 2308ΔOmp25 mutant. S2308 also activated p38 phosphorylation protein, extracellular-regulated protein kinases (ERK, and Jun-N-terminal kinase (JNK from MAPK signal pathway. 2308ΔOmp25 could not activate p38, ERK, and JNK branches. Immunohistochemistry experiments showed that S2308 was able to activate phosphorylation of p38 and ERK in BABL/c mice. However, 2308ΔOmp25 could weakly activate phosphorylation of p38 and ERK. These results suggest that Omp25 played an important role in the process of Brucella activation of the MAPK signal pathway.

  6. Mechanical properties and osteogenic activity of poly(l-lactide) fibrous membrane synergistically enhanced by chitosan nanofibers and polydopamine layer.

    Science.gov (United States)

    Liu, Hua; Li, Wenling; Wen, Wei; Luo, Binghong; Liu, Mingxian; Ding, Shan; Zhou, Changren

    2017-12-01

    To synergistically improve the mechanical properties and osteogenic activity of electrospinning poly(l-lactide) (PLLA) membrane, chitosan (CS) nanofibers were firstly introduced to prepare sub-micro and nanofibers interpenetrated PLLA/CS membrane, which was further surface modified with a polydopamine (PDA) layer to obtain PLLA/CS-PDA. Surface morphology, porosity, surface area and hydrophilicity of the obtained fibrous membranes were studied in detail. As compared to pure PLLA, the significant increase in the mechanical properties of the PLLA/CS, and especially of the PLLA/CS-PDA, was confirmed by tensile testing both in dry and wet states. Cells culture results indicated that both the PLLA/CS and PLLA/CS-PDA membranes, especially the latter, were more beneficial to adhesion, spreading and proliferation, as well as up-regulating alkaline phosphate activity and calcium deposition of MC3T3-E1 cells than PLLA membrane. Results suggested there was a synergistic effect of the CS nanofibers and PDA layer on the mechanical properties and osteogenic activity of PLLA membrane. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Membrane-Associated Transporter Protein (MATP Regulates Melanosomal pH and Influences Tyrosinase Activity.

    Directory of Open Access Journals (Sweden)

    Bum-Ho Bin

    Full Text Available The SLC45A2 gene encodes a Membrane-Associated Transporter Protein (MATP. Mutations of this gene cause oculocutaneous albinism type 4 (OCA4. However, the molecular mechanism of its action in melanogenesis has not been elucidated. Here, we discuss the role of MATP in melanin production. The SLC45A2 gene is highly enriched in human melanocytes and melanoma cell lines, and its protein, MATP, is located in melanosomes. The knockdown of MATP using siRNAs reduced melanin content and tyrosinase activity without any morphological change in melanosomes or the expression of melanogenesis-related proteins. Interestingly, the knockdown of MATP significantly lowered the melanosomal pH, as verified through DAMP analysis, suggesting that MATP regulates melanosomal pH and therefore affects tyrosinase activity. Finally, we found that the reduction of tyrosinase activity associated with the knockdown of MATP was readily recovered by copper treatment in the in vitro L-DOPA oxidase activity assay of tyrosinase. Considering that copper is an important element for tyrosinase activity and that its binding to tyrosinase depends on melanosomal pH, MATP may play an important role in regulating tyrosinase activity via controlling melanosomal pH.

  8. Membrane-Active Epithelial Keratin 6A Fragments (KAMPs Are Unique Human Antimicrobial Peptides with a Non-αβ Structure

    Directory of Open Access Journals (Sweden)

    Judy Tsz Ying Lee

    2016-11-01

    Full Text Available Antibiotic resistance is a pressing global health problem that threatens millions of lives each year. Natural antimicrobial peptides and their synthetic derivatives, including peptoids and peptidomimetics, are promising candidates as novel antibiotics. Recently, the C-terminal glycine-rich fragments of human epithelial keratin 6A were found to have bactericidal and cytoprotective activities. Here, we used an improved 2-dimensional NMR method coupled with a new protocol for structural refinement by low temperature simulated annealing to characterize the solution structure of these kerain-derived antimicrobial peptides (KAMPs. Two specific KAMPs in complex with membrane mimicking sodium dodecyl sulfate (SDS micelles displayed amphipathic conformations with only local bends and turns, and a central 10-residue glycine-rich hydrophobic strip that is central to bactericidal activity. To our knowledge, this is the first report of non-αβ structure for human antimicrobial peptides. Direct observation of Staphylococcus aureus and Pseudomonas aeruginosa by scanning and transmission electron microscopy showed that KAMPs deformed bacterial cell envelopes and induced pore formation. Notably, in competitive binding experiments, KAMPs demonstrated binding affinities to LPS and LTA that did not correlate with their bactericidal activities, suggesting peptide-LPS and peptide-LTA interactions are less important in their mechanisms of action. Moreover, immunoprecipitation of KAMPs-bacterial factor complexes indicated that membrane surface lipoprotein SlyB and intracellular machineries NQR sodium pump and ribosomes are potential molecular targets for the peptides. Results of this study improve our understanding of the bactericidal function of epithelial cytokeratin fragments, and highlight an unexplored class of human antimicrobial peptides, which may serve as non-αβ peptide scaffolds for the design of novel peptide-based antibiotics.

  9. Contribution of NADPH Oxidase to Membrane CD38 Internalization and Activation in Coronary Arterial Myocytes

    Science.gov (United States)

    Xu, Ming; Li, Xiao-Xue; Ritter, Joseph K.; Abais, Justine M.; Zhang, Yang; Li, Pin-Lan

    2013-01-01

    The CD38-ADP-ribosylcyclase-mediated Ca2+ signaling pathway importantly contributes to the vasomotor response in different arteries. Although there is evidence indicating that the activation of CD38-ADP-ribosylcyclase is associated with CD38 internalization, the molecular mechanism mediating CD38 internalization and consequent activation in response to a variety of physiological and pathological stimuli remains poorly understood. Recent studies have shown that CD38 may sense redox signals and is thereby activated to produce cellular response and that the NADPH oxidase isoform, NOX1, is a major resource to produce superoxide (O2·−) in coronary arterial myocytes (CAMs) in response to muscarinic receptor agonist, which uses CD38-ADP-ribosylcyclase signaling pathway to exert its action in these CAMs. These findings led us hypothesize that NOX1-derived O2·− serves in an autocrine fashion to enhance CD38 internalization, leading to redox activation of CD38-ADP-ribosylcyclase activity in mouse CAMs. To test this hypothesis, confocal microscopy, flow cytometry and a membrane protein biotinylation assay were used in the present study. We first demonstrated that CD38 internalization induced by endothelin-1 (ET-1) was inhibited by silencing of NOX1 gene, but not NOX4 gene. Correspondingly, NOX1 gene silencing abolished ET-1-induced O2·− production and increased CD38-ADP-ribosylcyclase activity in CAMs, while activation of NOX1 by overexpression of Rac1 or Vav2 or administration of exogenous O2·− significantly increased CD38 internalization in CAMs. Lastly, ET-1 was found to markedly increase membrane raft clustering as shown by increased colocalization of cholera toxin-B with CD38 and NOX1. Taken together, these results provide direct evidence that Rac1-NOX1-dependent O2·− production mediates CD38 internalization in CAMs, which may represent an important mechanism linking receptor activation with CD38 activity in these cells. PMID:23940720

  10. Levelling Profiles and a GPS Network to Monitor the Active Folding and Faulting Deformation in the Campo de Dalias (Betic Cordillera, Southeastern Spain)

    Science.gov (United States)

    Marín-Lechado, Carlos; Galindo-Zaldívar, Jesús; Gil, Antonio José; Borque, María Jesús; de Lacy, María Clara; Pedrera, Antonio; López-Garrido, Angel Carlos; Alfaro, Pedro; García-Tortosa, Francisco; Ramos, Maria Isabel; Rodríguez-Caderot, Gracia; Rodríguez-Fernández, José; Ruiz-Constán, Ana; de Galdeano-Equiza, Carlos Sanz

    2010-01-01

    The Campo de Dalias is an area with relevant seismicity associated to the active tectonic deformations of the southern boundary of the Betic Cordillera. A non-permanent GPS network was installed to monitor, for the first time, the fault- and fold-related activity. In addition, two high precision levelling profiles were measured twice over a one-year period across the Balanegra Fault, one of the most active faults recognized in the area. The absence of significant movement of the main fault surface suggests seismogenic behaviour. The possible recurrence interval may be between 100 and 300 y. The repetitive GPS and high precision levelling monitoring of the fault surface during a long time period may help us to determine future fault behaviour with regard to the existence (or not) of a creep component, the accumulation of elastic deformation before faulting, and implications of the fold-fault relationship. PMID:22319309

  11. Analysis of long-chain fatty acid binding activity in vesicles of the outer membrane generated from Escherchia coli

    International Nuclear Information System (INIS)

    Black, P.N.

    1987-01-01

    Escherichia coli transports long-chain fatty acids across the dual membrane by a high affinity, saturable, energy-dependent process. The fadL gene codes for an outer membrane protein which appears to act specifically as a long-chain fatty acid binding protein when fatty acid utilization is blocked by mutation. In an effort to understand the function of the fadL gene product, FLP, membranes have been isolated from fadL + and fadL - strains following osmotic lysis. Following isolation, total membranes were separated into inner and outer membrane fractions and assayed for long-chain fatty acid binding activity. Outer membrane vesicles were incubated 2-5 min at 37 0 C with 3 H oleate (C/sub 18:1/), cooled to 0 0 C, and centrifuged through a Lipidex 100 column for 3 min to remove the unbound fatty acid. The level of fatty acid binding was quantitated by scintillation counting of the eluate. Outer membrane vesicles generated from a fadL + strain bind 325 pmol fatty acid/mg protein whereas vesicles generated for a mutant strain bind 175 pmol fatty acid/mg protein. These data suggest that FLP acts at least as a long-chain fatty acid binding protein on the surface of the cell

  12. Characterization of antibacterial polyethersulfone membranes using the respiration activity monitoring system (RAMOS)

    NARCIS (Netherlands)

    Kochan, J.; Scheidle, M.; Erkel, J. van; Bikel, M.; Büchs, J.; Wong, J.E.; Melin, T.; Wessling, M.

    2012-01-01

    Membranes with antibacterial properties were developed using surface modification of polyethersulfone ultrafiltration membranes. Three different modification strategies using polyelectrolyte layer-by-layer (LbL) technique are described. The first strategy relying on the intrinsic antibacterial

  13. Continued Advancement of Supported Liquid Membranes for Carbon Dioxide Control in Extravehicular Activity Applications

    Science.gov (United States)

    Wickham, David T.; Gleason, Kevin J.; Engel, Jeffrey R.; Cowley, Scott W.; Chullen, Cinda

    2015-01-01

    The Development of a new, robust, portable life support system (PLSS) is currently a high NASA priority in order to support longer and safer extravehicular activity (EVA) missions that will be necessary as space travel extends to near-Earth asteroids and eventually Mars. One of the critical PLSS functions is maintaining the carbon dioxide (CO2) concentration in the suit at acceptable levels. The Metal Oxide (MetOx) canister has a finite CO2 adsorption capacity and therefore in order to extend mission times, the unit would have to be larger and heavier, which is undesirable; therefore new CO2 control technologies must be developed. While recent work has centered on the use of alternating sorbent beds that can be regenerated during the EVA, this strategy increases the system complexity and power consumption. A simpler approach is to use a membrane that vents CO2 to space but retains oxygen(O2). A membrane has many advantages over current technology: it is a continuous system with no theoretical capacity limit, it requires no consumables, and it requires no hardware for switching beds between absorption and regeneration. Conventional gas separation membranes do not have adequate selectivity for use in the PLSS, but the required performance could be obtained with a supported liquid membrane (SLM), which consists of a microporous film filled with a liquid that selectively reacts with CO2 over oxygen (O2). In a recently completed Phase II Small Business Innovative Research project, Reaction Systems developed a new reactive liquid that has effectively zero vapor pressure, making it an ideal candidate for use in an SLM. Results obtained with the SLM in a flat sheet configuration with representative pressures of CO2, O2, and water (H2O) have shown that the CO2 permeation rate and CO2/O2 selectivity requirements have been met. In addition, the SLM vents moisture to space very effectively. The SLM has also been prepared and tested in a hollow fiber form, which will be

  14. A Cell-Cell Fusion Assay to Assess Arenavirus Envelope Glycoprotein Membrane-Fusion Activity.

    Science.gov (United States)

    York, Joanne; Nunberg, Jack H

    2018-01-01

    For many viruses that enter their target cells through pH-dependent fusion of the viral and endosomal membranes, cell-cell fusion assays can provide an experimental platform for investigating the structure-function relationships that promote envelope glycoprotein membrane-fusion activity. Typically, these assays employ effector cells expressing the recombinant envelope glycoprotein on the cell surface and target cells engineered to quantitatively report fusion with the effector cell. In the protocol described here, Vero cells are transfected with a plasmid encoding the arenavirus envelope glycoprotein complex GPC and infected with the vTF7-3 vaccinia virus expressing the bacteriophage T7 RNA polymerase. These effector cells are mixed with target cells infected with the vCB21R-lacZ vaccinia virus encoding a β-galactosidase reporter under the control of the T7 promoter. Cell-cell fusion is induced upon exposure to low-pH medium (pH 5.0), and the resultant expression of the β-galactosidase reporter is quantitated using a chemiluminescent substrate. We have utilized this robust microplate cell-cell fusion assay extensively to study arenavirus entry and its inhibition by small-molecule fusion inhibitors.

  15. Regulated membrane remodeling by Mic60 controls formation of mitochondrial crista junctions.

    Science.gov (United States)

    Hessenberger, Manuel; Zerbes, Ralf M; Rampelt, Heike; Kunz, Séverine; Xavier, Audrey H; Purfürst, Bettina; Lilie, Hauke; Pfanner, Nikolaus; van der Laan, Martin; Daumke, Oliver

    2017-05-31

    The mitochondrial contact site and cristae organizing system (MICOS) is crucial for the formation of crista junctions and mitochondrial inner membrane architecture. MICOS contains two core components. Mic10 shows membrane-bending activity, whereas Mic60 (mitofilin) forms contact sites between inner and outer membranes. Here we report that Mic60 deforms liposomes into thin membrane tubules and thus displays membrane-shaping activity. We identify a membrane-binding site in the soluble intermembrane space-exposed part of Mic60. This membrane-binding site is formed by a predicted amphipathic helix between the conserved coiled-coil and mitofilin domains. The mitofilin domain negatively regulates the membrane-shaping activity of Mic60. Binding of Mic19 to the mitofilin domain modulates this activity. Membrane binding and shaping by the conserved Mic60-Mic19 complex is crucial for crista junction formation, mitochondrial membrane architecture and efficient respiratory activity. Mic60 thus plays a dual role by shaping inner membrane crista junctions and forming contact sites with the outer membrane.

  16. Regulation of mitogen-activated protein kinase pathways by the plasma membrane Na+/H+ exchanger, NHE1

    DEFF Research Database (Denmark)

    Pedersen, Stine Helene Falsig; Darborg, Barbara Vasek; Rentsch, Maria Louise

    2006-01-01

    activity is regulated by a three-tiered phosphorelay system, which is in turn regulated by a complex network of signaling events and scaffolding proteins. The ubiquitous plasma membrane Na(+)/H(+) exchanger NHE1 is activated by, and implicated in, the physiological/pathophysiological responses to many...

  17. New Insight Into the Roles of Membrane Microdomains in Physiological Activities of Fungal Cells.

    Czech Academy of Sciences Publication Activity Database

    Malínský, Jan; Opekarová, Miroslava

    2016-01-01

    Roč. 325, mar. (2016), s. 119-180 ISSN 1937-6448 R&D Projects: GA ČR(CZ) GA15-10641S Institutional support: RVO:68378041 Keywords : membrane microdomain * membrane structure * fungi * membrane contact sites Subject RIV: EA - Cell Biology Impact factor: 3.752, year: 2015

  18. Probing simultaneously membrane dynamics and protein activity in suspended bilayers in a microfluidic format

    NARCIS (Netherlands)

    Schulze Greiving-Stimberg, Verena Carolin; Bomer, Johan G.; de Boer, Hans L.; van den Berg, Albert; le Gac, Severine

    2014-01-01

    Membrane dynamics affect the structure and function of ion channels, a point that deserves more attention while studying membrane proteins. One important factor in the local lipidic environment of the ion channels, is the membrane fluidity which is directly connected to the free diffusion and

  19. Hearts lacking plasma membrane KATPchannels display changes in basal aerobic metabolic substrate preference and AMPK activity.

    Science.gov (United States)

    Youssef, Nermeen; Campbell, Scott; Barr, Amy; Gandhi, Manoj; Hunter, Beth; Dolinsky, Vernon; Dyck, Jason R B; Clanachan, Alexander S; Light, Peter E

    2017-09-01

    Cardiac ATP-sensitive K + (K ATP ) channels couple changes in cellular metabolism to membrane excitability and are activated during metabolic stress, although under basal aerobic conditions, K ATP channels are thought to be predominately closed. Despite intense research into the roles of K ATP channels during metabolic stress, their contribution to aerobic basal cardiac metabolism has not been previously investigated. Hearts from Kir6.2 +/+ and Kir6.2 -/- mice were perfused in working mode, and rates of glycolysis, fatty acid oxidation, and glucose oxidation were measured. Changes in activation/expression of proteins regulating metabolism were probed by Western blot analysis. Despite cardiac mechanical function and metabolic efficiency being similar in both groups, hearts from Kir6.2 -/- mice displayed an approximately twofold increase in fatty acid oxidation and a 0.45-fold reduction in glycolytic rates but similar glucose oxidation rates compared with hearts from Kir6.2 +/+ mice. Kir6.2 -/- hearts also possessed elevated levels of activated AMP-activated protein kinase (AMPK), higher glycogen content, and reduced mitochondrial density. Moreover, activation of AMPK by isoproterenol or diazoxide was significantly blunted in Kir6.2 -/- hearts. These data indicate that K ATP channel ablation alters aerobic basal cardiac metabolism. The observed increase in fatty acid oxidation and decreased glycolysis before any metabolic insult may contribute to the poor recovery observed in Kir6.2 -/- hearts in response to exercise or ischemia-reperfusion injury. Therefore, K ATP channels may play an important role in the regulation of cardiac metabolism through AMPK signaling. NEW & NOTEWORTHY In this study, we show that genetic ablation of plasma membrane ATP-sensitive K + channels results in pronounced changes in cardiac metabolic substrate preference and AMP-activated protein kinase activity. These results suggest that ATP-sensitive K + channels may play a novel role in

  20. Improving the performance of an aerobic membrane bioreactor (MBR) treating pharmaceutical wastewater with powdered activated carbon (PAC) addition.

    Science.gov (United States)

    Kaya, Yasemin; Bacaksiz, A Murat; Golebatmaz, Ugur; Vergili, Ilda; Gönder, Z Beril; Yilmaz, Gulsum

    2016-04-01

    In this study, the effects of organic loading rate (OLR) and the addition of powdered activated carbon (PAC) on the performance and membrane fouling of MBR were conducted to treat real pharmaceutical process wastewater. Over 145 days of operation, the MBR system was operated at OLRs ranging from 1 to 2 kg COD m(-3) day(-1) without sludge wasting. The addition of PAC provided an improvement in the flux, despite an increase in the OLR:PAC ratio. The results demonstrated that the hybrid PAC-MBR system maintained a reduced amount of membrane fouling and steadily increased the removal performance of etodolac. PAC addition reduced the deposition of extracellular polymeric substance and organic matter on the membrane surface and resulted an increase in COD removal even at higher OLRs with low PAC addition. Membrane fouling mechanisms were investigated using combined adsorption fouling models. Modified fouling index values and normalized mass transfer coefficient values indicated that predominant fouling mechanism was cake adsorption.

  1. An ABC-transporter and an outer membrane lipoprotein participate in posttranslational activation of type VI secretion in Pseudomonas aeruginosa

    Science.gov (United States)

    Casabona, Maria G.; Silverman, Julie M.; Sall, Khady M.; Boyer, Frédéric; Couté, Yohann; Poirel, Jessica; Grunwald, Didier; Mougous, Joseph D.; Elsen, Sylvie; Attree, Ina

    2012-01-01

    Pseudomonas aeruginosa is capable of injecting protein toxins into other bacterial cells through one of its three type VI secretion systems (T6SS). The activity of this T6SS is tightly regulated on the posttranslational level by phosphorylation-dependent and -independent pathways. The phosphorylation-dependent pathway consists of a Thr kinase/phosphatase pair (PpkA/PppA) that acts on a forkhead domain-containing protein Fha1, and a periplasmic protein, TagR, that positively regulates PpkA. In the present work, we biochemically and functionally characterize three additional proteins of the phosphorylation-dependent regulatory cascade that controls T6S activation: TagT, TagS and TagQ. We show that similar to TagR, these proteins act upstream of the PpkA/PppA checkpoint and influence phosphorylation of Fha1 and export of Hcp1 and Tse1. Localization studies demonstrate that TagQ is an outer membrane lipoprotein and TagR is associated with the outer membrane. Consistent with their homology to lipoprotein outer membrane localization (Lol) components, TagT and TagS form a stable inner membrane complex with ATPase activity. However, we find that outer membrane association of T6SS lipoproteins TagQ and TssJ1, and TagR, is unaltered in a ΔtagTS background. Notably, we found that TagQ is indispensible for anchoring of TagR to the outer membrane fraction. As T6S-dependent fitness of P. aeruginosa requires TagT, S, R and Q, we conclude that these proteins likely participate in a trans-membrane signaling pathway that promotes H1-T6SS activity under optimal environmental conditions. PMID:22765374

  2. Comparative study of the active cadmium efflux systems operating at the plasma membrane and tonoplast of cucumber root cells.

    Science.gov (United States)

    Migocka, Magdalena; Papierniak, Anna; Kosatka, Ewelina; Klobus, Grazyna

    2011-10-01

    The strategies developed by plants to avoid the toxicity of cadmium (Cd) and other heavy metals involve active sequestration of metals into the apoplast and vacuoles. The protein systems excluding heavy metals from the cell cytosol localize to the plasma membrane and tonoplast and are energized either by ATP or by the electrochemical gradient generated by H(+)-ATPase or by V-ATPase and pyrophosphatase (PPase), respectively. In this work, a comparative study on the contribution of both the plasma membrane and tonoplast in the active detoxification of plant cells after treatment with Cd was performed. The studies using plants treated and untreated with Cd reveal that both, H(+)-coupled and MgATP-driven efflux of Cd across plasma membranes and tonoplast is markedly stimulated in the presence of Cd in the environment. Previous studies on plasma-membrane localized H(+)-coupled Cd efflux together with the present data demonstrating tonoplast H(+)/Cd(2+) antiport activity suggest that H(+)-coupled secondary transport of Cd displays a lower affinity for Cd when compared with Cd primary pumps driven by MgATP. In addition, it is shown that MgATP-energized Cd efflux across both membranes is significantly enhanced by cysteine, dithiothreitol, and glutathione. These results suggest that Cd is excluded from the cytosol through an energy-dependent system as a free ion as well as a complexed form. Although both membranes contribute in the active exclusion of ionized and complexed Cd from the cytosol, the overall calculation of Cd accumulation in the everted plasma membranes and vacuolar vesicles suggests that the tonoplast and vacuole have a major function in Cd efflux from the cytosol in the roots of cucumber subjected to Cd stress.

  3. An ABC transporter and an outer membrane lipoprotein participate in posttranslational activation of type VI secretion in Pseudomonas aeruginosa.

    Science.gov (United States)

    Casabona, Maria G; Silverman, Julie M; Sall, Khady M; Boyer, Frédéric; Couté, Yohann; Poirel, Jessica; Grunwald, Didier; Mougous, Joseph D; Elsen, Sylvie; Attree, Ina

    2013-02-01

    Pseudomonas aeruginosa is capable of injecting protein toxins into other bacterial cells through one of its three type VI secretion systems (T6SSs). The activity of this T6SS is tightly regulated on the posttranslational level by phosphorylation-dependent and -independent pathways. The phosphorylation-dependent pathway consists of a Threonine kinase/phosphatase pair (PpkA/PppA) that acts on a forkhead domain-containing protein, Fha1, and a periplasmic protein, TagR, that positively regulates PpkA. In the present work, we biochemically and functionally characterize three additional proteins of the phosphorylation-dependent regulatory cascade that controls T6S activation: TagT, TagS and TagQ. We show that similar to TagR, these proteins act upstream of the PpkA/PppA checkpoint and influence phosphorylation of Fha1 and, apparatus assembly and effector export. Localization studies demonstrate that TagQ is an outer membrane lipoprotein and TagR is associated with the outer membrane. Consistent with their homology to lipoprotein outer membrane localization (Lol) components, TagT and TagS form a stable inner membrane complex with ATPase activity. However, we find that outer membrane association of T6SS lipoproteins TagQ and TssJ1, and TagR, is unaltered in a ΔtagTS background. Notably, we found that TagQ is indispensible for anchoring of TagR to the outer membrane fraction. As T6S-dependent fitness of P. aeruginosa requires TagT, S, R and Q, we conclude that these proteins likely participate in a trans-membrane signalling pathway that promotes H1-T6SS activity under optimal environmental conditions. © 2012 Society for Applied Microbiology and Blackwell Publishing Ltd.

  4. Deformation of slopes as a cause of underground mining activities: three case studies from Ostrava-Karviná coal field (Czech Republic).

    Science.gov (United States)

    Marschalko, Marian; Yilmaz, Işık; Bednárik, Martin; Kubečka, Karel

    2012-11-01

    Underground mining activities may potentially play a role on the initiation and reactivation of the slope movements. However, an adequate attention has not yet been paid to these problems; in this study, the possible influence of present and former mining activities on the selected set of model slope deformations in the Ostrava-Karviná Coalfield (Opliji, Repiste and Orlova Lazy District) was analysed and a methodology for their observation for application to similar conditions and influence was described. Isocatabase maps, terrain deformation parameters calculated for the point lying on the slope deformation surface, length measurement by zone extensometer and dilatometer measurement in cracks was also provided for evaluation of the underground mining impact. It was found that inclinations of both boreholes were evidence of underground mining impact, and localization of inclinometer measurement on boreholes in the active part as well as in the near vicinity was very important as an important result of this study. Analysis of underground mining activity influence on model localities in relation to performed mining operations, subsidence and other influences on the ground surface was also determined. Thus, the study will contribute to a more objective knowledge of these problems of interest for the professional public and also for the state administration to solve problems associated with the utilisation and settlement of such affected areas.

  5. Gravity changes and deformation at Kīlauea Volcano, Hawaii, associated with summit eruptive activity, 2009-2012

    Science.gov (United States)

    Bagnardi, Marco; Poland, Michael P.; Carbone, Daniele; Baker, Scott; Battaglia, Maurizio; Amelung, Falk

    2014-01-01

    Analysis of microgravity and surface displacement data collected at the summit of Kīlauea Volcano, Hawaii (USA), between December 2009 and November 2012 suggests a net mass accumulation at ~1.5 km depth beneath the northeast margin of Halema‘uma‘u Crater, within Kīlauea Caldera. Although residual gravity increases and decreases are accompanied by periods of uplift and subsidence of the surface, respectively, the volume change inferred from the modeling of interferometric synthetic aperture radar deformation data can account for only a small portion (as low as 8%) of the mass addition responsible for the gravity increase. We propose that since the opening of a new eruptive vent at the summit of Kīlauea in 2008, magma rising to the surface of the lava lake outgasses, becomes denser, and sinks to deeper levels, replacing less dense gas-rich magma stored in the Halema‘uma‘u magma reservoir. In fact, a relatively small density increase (<200 kg m−3) of a portion of the reservoir can produce the positive residual gravity change measured during the period with the largest mass increase, between March 2011 and November 2012. Other mechanisms may also play a role in the gravity increase without producing significant uplift of the surface, including compressibility of magma, formation of olivine cumulates, and filling of void space by magma. The rate of gravity increase, higher than during previous decades, varies through time and seems to be directly correlated with the volcanic activity occurring at both the summit and the east rift zone of the volcano.

  6. Removal of emerging contaminants by simultaneous application of membrane ultrafiltration, activated carbon adsorption, and ultrasound irradiation.

    Science.gov (United States)

    Secondes, Mona Freda N; Naddeo, Vincenzo; Belgiorno, Vincenzo; Ballesteros, Florencio

    2014-01-15

    Advanced wastewater treatment is necessary to effectively remove emerging contaminants (ECs) with chronic toxicity, endocrine disrupting effects, and the capability to induce the proliferation of highly resistant microbial strains in the environment from before wastewater disposal or reuse. This paper investigates the efficiency of a novel hybrid process that applies membrane ultrafiltration, activated carbon adsorption, and ultrasound irradiation simultaneously to remove ECs. Diclofenac, carbamazepine, and amoxicillin are chosen for this investigation because of their assessed significant environmental risks. Removal mechanisms and enhancement effects are analysed in single and combined processes. The influence of adsorbent dose and ultrasonic frequency to EC removal are also investigated. Results suggest that adsorption is probably the main removal mechanism and is affected by the nature of ECs and the presence of other components in the mixture. Almost complete removals are achieved in the hybrid process for all ECs. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. Hemodialysis leukopenia. Pulmonary vascular leukostasis resulting from complement activation by dialyzer cellophane membranes.

    Science.gov (United States)

    Craddock, P R; Fehr, J; Dalmasso, A P; Brighan, K L; Jacob, H S

    1977-01-01

    Acute leukopenia occurs in all patients during the first hour of hemodialysis with cellophanemembrane equipment. This transient cytopenia specifically involves granulocytes and monocytes, cells which share plasma membrane reactivity towards activated complement components. The present studies document that complement is activated during exposure of plasma to dialyzer cellophane, and that upon reinfusion of this plasma into the venous circulation, granulocyte and monocyte entrapment in the pulmonary vasculature is induced. During early dialysis, conversion of both C3 and factor B can be demonstrated in plasma as it leaves the dialyzer. Moreover, simple incubation of human plasma with dialyzer cellophane causes conversion of C3 and factor B, accompanied by depletion of total hemolytic complement and C3 but sparing of hemolytic C1. Reinfusion of autologous, cellophane-incubated plasma into rabbits produces selective granulocytopenia and monocytopenia identical to that seen in dialyzed patients. Lungs from such animals reveal striking pulmonary vessel engorgement with granulocytes. The activated complement component(s) responsible for leukostasis has an approximate molecular weight of 7,000-20,000 daltons. Since it is generated in C2-deficient plasma and is associated with factor B conversion, it is suggested that activation of complement by dialysis is predominantly through the altermative pathway. Images PMID:856872

  8. Effluent quality of a conventional activated sludge and a membrane bioreactor system treating hospital wastewater

    International Nuclear Information System (INIS)

    Pauwels, B.; Ngwa, F.; Deconinck, S.; Verstraete, W.

    2005-01-01

    Two lab scale wastewater treatment plants treating hospital wastewater in parallel were compared in terms of performance characteristics. One plant consisted of a conventional activated sludge system (CAS) and comprised In anoxic and aerobic compartment followed by a settling tank with recycle loop. The second pilot plant was a -late membrane bioreactor (MBR). The wastewater as obtained from the hospital had a variable COD (Chemical Oxygen Demand) ranging from 250 to 2300 mg/L. Both systems were operated at a similar hydraulic residence time of 12 hours. The reference conventional activated sludge system did not meet the regulatory standard for effluent COD of 125 mg /L most of the time. Its COD removal efficiency was 88%. The plate MBR delivered an effluent with a COD value of 50 mg/L or less, and attained an efficiency of 93%. The effluent contained no suspended particles. In addition, the MBR resulted in consistent operational parameters with a flux remaining around 8 -10 L/m/sup 2/.h and a trans membrane pressure <0.1 bar without the need for backwash or chemical cleaning. The CAS and the MBR system performed equally good in terms of TAN removal and EE2 removal. The CAS system typically decreased bacterial groups for about 1 log unit, whereas the MBR decreased these groups for about 3 log units. Enterococci were decreased below the detection limit in the MBR and indicator organisms such as fecal coliforms were decreased for 1.4 log units in the CAS system compared to a 3.6 log removal in the MBR. (author)

  9. Functional activation of G-proteins coupled with muscarinic acetylcholine receptors in rat brain membranes.

    Science.gov (United States)

    Odagaki, Yuji; Kinoshita, Masakazu; Toyoshima, Ryoichi

    2014-01-01

    The functional activation of Gi/o proteins coupled to muscarinic acetylcholine receptors (mAChRs) was investigated with the conventional guanosine-5'-O-(3-[(35)S]thio) triphosphate ([(35)S]GTPγS) binding assay in rat brain membranes. The most efficacious stimulation elicited by acetylcholine or carbachol (CCh) was obtained in striatal membranes. The pharmacological properties of mAChR-mediated [(35)S]GTPγS binding determined with a series of muscarinic agonists and antagonists were almost identical among the three brain regions investigated, i.e., cerebral cortex, hippocampus, and striatum, except for the apparent partial agonist effects of (αR)-α-cyclopentyl-α-hydroxy-N-[1-(4-methyl-3-pentenyl)-4-piperidinyl]benzeneacetamide fumarate (J 104129) observed only in the hippocampus, but not in the other two regions. Among the muscarinic toxins investigated, only MT3 attenuated CCh-stimulated [(35)S] GTPγS binding. The highly selective allosteric potentiator at the M4 mAChR subtype, 3-amino-N-[(4-chlorophenyl)methyl]-4,6-dimethylthieno[2,3-b]pyridine-2-carboxamide (VU 10010), shifted the concentration-response curve for CCh leftwards as well as upwards. On the other hand, neither thiochrome nor brucine N-oxide was effective. The increases induced by CCh and 5-HT were essentially additive, though not completely, indicating that the mAChRs and 5-HT1A receptors were coupled independently to distinct pools of Gi/o proteins. Collectively, all of the data suggest that functional activation of Gi/o proteins coupled to mAChRs, especially the M4 subtype, is detectable by means of CCh-stimulated [(35)S]GTPγS binding assay in rat discrete brain regions.

  10. Interplay between membrane elasticity and active cytoskeleton forces regulates the aggregation dynamics of the immunological synapse

    Science.gov (United States)

    Dharan, Nadiv; Farago, Oded

    Adhesion between a T cell and an antigen presenting cell is achieved by TCR-pMHC and LFA1-ICAM1 protein complexes. These segregate to form a special pattern, known as the immunological synapse (IS), consisting of a central quasi-circular domain of TCR-pMHC bonds surrounded by a peripheral domain of LFA1-ICAM1 complexes. Insights gained from imaging studies had led to the conclusion that the formation of the central adhesion domain in the IS is driven by active (ATP-driven) mechanisms. Recent studies, however, suggested that passive (thermodynamic) mechanisms may also play an important role in this process. Here, we present a simple physical model, taking into account the membrane-mediated thermodynamic attraction between the TCR-pMHC bonds and the effective forces that they experience due to ATP-driven actin retrograde flow and transport by dynein motor proteins. Monte Carlo simulations of the model exhibit a good spatio-temporal agreement with the experimentally observed pattern evolution of the TCR-pMHC microclusters. The agreement is lost when one of the aggregation mechanisms is "muted", which helps to identify the respective roles in the process. We conclude that actin retrograde flow drives the centripetal motion of TCR-pMHC bonds, while the membrane-mediated interactions facilitate microcluster formation and growth. In the absence of dynein motors, the system evolves into a ring-shaped pattern, which highlights the role of dynein motors in the formation of the final concentric pattern. The interplay between the passive and active mechanisms regulates the rate of the accumulation process, which in the absence of one them proceeds either too quickly or slowly.

  11. Antimicrobial properties and membrane-active mechanism of a potential α-helical antimicrobial derived from cathelicidin PMAP-36.

    Directory of Open Access Journals (Sweden)

    Yinfeng Lv

    Full Text Available Antimicrobial peptides (AMPs, which present in the non-specific immune system of organism, are amongst the most promising candidates for the development of novel antimicrobials. The modification of naturally occurring AMPs based on their residue composition and distribution is a simple and effective strategy for optimization of known AMPs. In this study, a series of truncated and residue-substituted derivatives of antimicrobial peptide PMAP-36 were designed and synthesized. The 24-residue truncated peptide, GI24, displayed antimicrobial activity comparable to the mother peptide PMAP-36 with MICs ranging from 1 to 4 µM, which is lower than the MICs of bee venom melittin. Although GI24 displayed high antimicrobial activity, its hemolytic activity was much lower than melittin, suggesting that GI24 have optimal cell selectivity. In addition, the crucial site of GI24 was identified through single site-mutation. An amino acid with high hydrophobicity at position 23 played an important role in guaranteeing the high antimicrobial activity of GI24. Then, lipid vesicles and whole bacteria were employed to investigate the membrane-active mechanisms. Membrane-simulating experiments showed that GI24 interacted strongly with negatively charged phospholipids and weakly with zwitterionic phospholipids, which corresponded well with the data of its biological activities. Membrane permeabilization and flow cytometry provide the evidence that GI24 killed microbial cells by permeabilizing the cell membrane and damaging membrane integrity. GI24 resulted in greater cell morphological changes and visible pores on cell membrane as determined using scanning electron microscopy (SEM and transmission electron microscope (TEM. Taken together, the peptide GI24 may provide a promising antimicrobial agent for therapeutic applications against the frequently-encountered bacteria.

  12. SARS-coronavirus replication/transcription complexes are membrane-protected and need a host factor for activity in vitro.

    Directory of Open Access Journals (Sweden)

    Martijn J van Hemert

    2008-05-01

    Full Text Available SARS-coronavirus (SARS-CoV replication and transcription are mediated by a replication/transcription complex (RTC of which virus-encoded, non-structural proteins (nsps are the primary constituents. The 16 SARS-CoV nsps are produced by autoprocessing of two large precursor polyproteins. The RTC is believed to be associated with characteristic virus-induced double-membrane structures in the cytoplasm of SARS-CoV-infected cells. To investigate the link between these structures and viral RNA synthesis, and to dissect RTC organization and function, we isolated active RTCs from infected cells and used them to develop the first robust assay for their in vitro activity. The synthesis of genomic RNA and all eight subgenomic mRNAs was faithfully reproduced by the RTC in this in vitro system. Mainly positive-strand RNAs were synthesized and protein synthesis was not required for RTC activity in vitro. All RTC activity, enzymatic and putative membrane-spanning nsps, and viral RNA cosedimented with heavy membrane structures. Furthermore, the pelleted RTC required the addition of a cytoplasmic host factor for reconstitution of its in vitro activity. Newly synthesized subgenomic RNA appeared to be released, while genomic RNA remained predominantly associated with the RTC-containing fraction. RTC activity was destroyed by detergent treatment, suggesting an important role for membranes. The RTC appeared to be protected by membranes, as newly synthesized viral RNA and several replicase/transcriptase subunits were protease- and nuclease-resistant and became susceptible to degradation only upon addition of a non-ionic detergent. Our data establish a vital functional dependence of SARS-CoV RNA synthesis on virus-induced membrane structures.

  13. Properties of the cyanobacterial coupling factor ATPase from Spirulina platensis. II. Activity of the purified and membrane-bound enzymes.

    Science.gov (United States)

    Hicks, D B; Yocum, C F

    1986-02-15

    Cyanobacterial (Spirulina platensis) photosynthetic membranes and isolated F1 ATPase were characterized with respect to ATP activity. The following results indicate that the regulation of expression of ATPase activity in Spirulina platensis is similar to that found in chloroplasts: the ATPase activity of Spirulina membranes and isolated F1 ATPase is mostly latent, a characteristic of chloroplast ATPase activity; treatments that elicit ATPase activity in higher plant chloroplast thylakoids and isolated chloroplast coupling factor (CF1) greatly stimulate the activity of Spirulina membranes and F1, and the cation specificity of chloroplast ATPase activity, e. g., light-induced membrane activity that is magnesium dependent and trypsin-activated CF1 activity that is calcium dependent, is also observed in Spirulina. Thus, an 8- to 15-fold increase in specific activity (to 13-15 mumol Pi min-1 mg chl-1) is obtained when Spirulina membranes are treated with trypsin (CaATPase) or with methanol (MgATPase): a light-induced, dithiothreitol-dependent MgATPase activity is also found in the membranes. Purified Spirulina F1 is a CaATPase when activated with trypsin (endogenous activity increases from 4 to 27-37 mumol Pi min-1 mg protein-1) or with dithiothreitol (5.6 mumol Pi min-1 mg-1), but a MgATPase when assayed with methanol (18-20 mumol Pi min-1 mg-1). The effects of varying calcium and ATP concentrations on the kinetics of trypsin-induced CaATPase activity of Spirulina F1 were examined. When the calcium concentration is varied at constant ATP concentration, the velocity plot shows a marked sigmoidicity. By varying Ca-ATP metal-nucleotide complex concentration at constant concentrations of free calcium or ATP, it is shown that the sigmoidicity is due to the effect of free ATP, which changes the Hill constant to 1.6 from 1.0 observed when the free calcium concentration is kept constant at 5 mM. Therefore not only is ATP an inhibitor but it is also an allosteric effector of

  14. The intrinsically disordered tails of PTEN and PTEN-L have distinct roles in regulating substrate specificity and membrane activity

    Science.gov (United States)

    Masson, Glenn R.; Perisic, Olga; Burke, John E.; Williams, Roger L.

    2015-01-01

    Phosphatase and tensin homologue deleted on chromosome 10 (PTEN) is a lipid and protein phosphatase, and both activities are necessary for its role as a tumour suppressor. PTEN activity is controlled by phosphorylation of its intrinsically disordered C-terminal tail. A recently discovered variant of PTEN, PTEN-long (PTEN-L), has a 173-residue N-terminal extension that causes PTEN-L to exhibit unique behaviour, such as movement from one cell to another. Using hydrogen/deuterium exchange mass spectrometry (HDX–MS) and biophysical assays, we show that both the N-terminal extension of PTEN-L and C-terminal tail of PTEN affect the phosphatase activity using unique mechanisms. Phosphorylation of six residues in the C-terminal tail of PTEN results in auto-inhibitory interactions with the phosphatase and C2 domains, effectively blocking both the active site and the membrane-binding interface of PTEN. Partially dephosphorylating PTEN on pThr366/pSer370 results in sufficient exposure of the active site to allow a selective activation for soluble substrates. Using HDX–MS, we identified a membrane-binding element in the N-terminal extension of PTEN-L, termed the membrane-binding helix (MBH). The MBH radically alters the membrane binding mechanism of PTEN-L compared with PTEN, switching PTEN-L to a ‘scooting’ mode of catalysis from the ‘hopping’ mode that is characteristic of PTEN. PMID:26527737

  15. Eggshell membrane hydrolyzates activate NF-κB in vitro: possible implications for in vivo efficacy

    Directory of Open Access Journals (Sweden)

    Ruff KJ

    2015-02-01

    Full Text Available Kevin J Ruff,1 Paul L Durham,2 Austin O’Reilly,2 F Daniel Long1 1ESM Technologies, LLC, Carthage, MO, USA; 2Center for Biomedical and Life Sciences, Missouri State University, Springfield, MO, USA Purpose: Eggshell membrane (ESM has been shown to contain naturally occurring bioactive components, and biological activities such as reducing proinflammatory cytokines, liver fibrosis, and joint pain in osteoarthritis sufferers have also been reported for ESM matrix as a whole. Nuclear factor kappa-light-chain-enhancer of activated B-cells (NF-κB is a signaling protein found in the cytoplasm of nearly all human and animal cell types and is a primary regulator of immune function. The studies reported herein were designed to investigate the possible role that NF-κB activity might play in the reported biological activities of ESM. Methods: Three ESM hydrolyzates produced via fermentation, enzymatic, or chemical hydrolysis were evaluated in vitro in either human peripheral blood mononuclear cell or THP-1 (human leukemic monocyte cell cultures for NF-κB activity following 4-hour exposure. The hydrolyzates were compared with untreated control cells or cells incubated with lipopolysaccharide or ascorbic acid. The source of ESM activity was also evaluated. Results: NF-κB levels were increased above levels found in untreated cells at all three dilutions (1:100, 1:1,000, and 1:10,000 for the fermentation hydrolyzate of ESM (ESM-FH (P=0.021, P=0.020, P=0.009, respectively in peripheral blood mononuclear cells. The enzymatic hydrolyzate of ESM (ESM-EH also produced statistically significant levels of activated NF-κB at the 1:100 and 1:1,000 dilutions (P=0.004, P=0.006, respectively but fell just shy of significance at the 1:10,000 dilution (P=0.073. Similarly, ESM-FH (P=0.021, P=0.002 and ESM-EH (P=0.007, P=0.007 activated NF-κB in THP-1 cells at 1:1,000 and 1:10,000 dilutions, respectively. The chemical hydrolyzate of ESM (ESM-CH showed statistically

  16. Phenylarsine Oxide Inhibits the Fusicoccin-Induced Activation of Plasma Membrane H+-ATPase1

    Science.gov (United States)

    Olivari, Claudio; Albumi, Cristina; Pugliarello, Maria Chiara; De Michelis, Maria Ida

    2000-01-01

    To investigate the mechanism by which fusicoccin (FC) induces the activation of the plasma membrane (PM) H+-ATPase, we used phenylarsine oxide (PAO), a known inhibitor of protein tyrosine-phosphatases. PAO was supplied in vivo in the absence or presence of FC to radish (Raphanus sativus L.) seedlings and cultured Arabidopsis cells prior to PM extraction. Treatment with PAO alone caused a slight decrease of PM H+-ATPase activity and, in radish, a decrease of PM-associated 14-3-3 proteins. When supplied prior to FC, PAO drastically inhibited FC-induced activation of PM H+-ATPase, FC binding to the PM, and the FC-induced increase of the amount of 14-3-3 associated with the PM. On the contrary, PAO was completely ineffective on all of the above-mentioned parameters when supplied after FC. The H+-ATPase isolated from PAO-treated Arabidopsis cells maintained the ability to respond to FC if supplied with exogenous, nonphosphorylated 14-3-3 proteins. Altogether, these results are consistent with a model in which the dephosphorylated state of tyrosine residues of a protein(s), such as 14-3-3 protein, is required to permit FC-induced association between the 14-3-3 protein and the PM H+-ATPase. PMID:10677439

  17. Antioxidant activity of capsaicin on radiation-induced oxidation of murine hepatic mitochondrial membrane preparation

    Directory of Open Access Journals (Sweden)

    Gangabhagirathi R

    2015-06-01

    Full Text Available Ramachandran Gangabhagirathi,1 Ravi Joshi,2 1Bioorganic Division, 2Radiation and Photochemistry Division, Bhabha Atomic Research Center, Trombay, Mumbai, India Abstract: Capsaicin is the major capsaicinoid in chili peppers and is widely used as a spice. It is also used for topical applications in cases of peripheral neuropathy. The present study deals with its role in modulation of gamma radiation-induced damages of the biochemical constituents of rat liver mitochondrial membrane (RLM preparation. The extent of lipid hydroperoxide formation, depletion in protein thiols, and formation of protein carbonyls have been biochemically assessed in the presence of varying concentrations of capsaicin in RLM. Decrease in the activities of the important antioxidant enzyme superoxide dismutase, which is involved in the scavenging of free radicals, and the mitochondrial marker enzyme succinate dehydrogenase have been also looked into. Capsaicin has been found to efficiently inhibit radiation-induced biochemical alterations, namely lipid peroxidation and protein oxidation. It also significantly prevented radiation-induced loss in the activity of antioxidant enzyme and the important endogenous antioxidant glutathione. The study suggests that capsaicin can act as an antioxidant and radioprotector in physiological systems. Keywords: capsaicin, gamma radiation, radioprotection, lipid peroxidation, protein oxidation, enzyme activity

  18. Scintigraphic evaluation of the osteoblastic activity of rabbit tibial defects after HYAFF11 membrane application.

    Science.gov (United States)

    Mermerkaya, Musa Uğur; Doral, Mahmut Nedim; Karaaslan, Fatih; Huri, Gazi; Karacavuş, Seyhan; Kaymaz, Burak; Alkan, Erkan

    2016-05-03

    An unfavorable condition for bone healing is the presence of bone defects. Under such conditions, a material can play a role to cover fractured or defective bone. Technological advances now allow for the use of such material. Hyalonect(®) (Fidia Advanced Biopolymers SLR, Italy), a novel membrane comprising knitted fibers of esterified hyaluronan (HYAFF11) can be used to cover fractured or grafted bone and can also serve as a scaffold to keep osteoprogenitor cells in place. The aim of this study was to compare osteoblastic activity by the use of scintigraphic methods in defective rabbit tibias during early-phase bone healing with or without a hyaluronan-based mesh. Two groups (A and B) of New Zealand albino rabbits were used; each group included 10 animals. Operations on all rabbits were performed under general anesthesia. We also resected 10-mm bone segments from each animal's tibial diaphysis. After resection, tibias with defects were fixed using Kirschner wires. In group A, no hyaluronan-based mesh was used. In group B, tibial segmental defects were enclosed with a hyaluronan-based mesh. The rabbits were followed up for 4 weeks postoperatively, after which bone scintigraphic studies were performed on each animal to detect and compare osteoblastic activity. The mean count in the fracture side of the hyaluronan-based mesh group was significantly higher compared to that of the group A (p = 0.019). However, there was no significant difference between group B and control rabbits with respect to the mean count on the intact bone side (p = 0.437). The bone defect (fracture)/intact bone mean count ratio was significantly higher in group B compared to group A (p = 0.008). A hyaluronan-based mesh plays a role in promoting osteoblastic activity. Hyalonect(®) is suitable for restoring tissue continuity whenever the periosteal membrane is structurally impaired or inadequate. Our results demonstrated that, during early-phase bone healing, osteoblastic activity

  19. Ultrasonic irradiation to modify the functionalized bionanocomposite in sulfonated polybenzimidazole membrane for fuel cells applications and antibacterial activity.

    Science.gov (United States)

    Esmaeilzade, Banafshe; Esmaielzadeh, Sheida; Ahmadizadegan, Hashem

    2018-04-01

    In this article the new proton exchange membranes were prepared from sulfonated polybenzimidazole (s-PBI) and various amounts of sulfonated titania/cellulose nanohybrids (titania/cellulose-SO 3 H) via ultrasonic waves. The ultrasonic irradiation effectively changes the rheology and the glass transition temperature and the crystallinity of the composite polymer. Ultrasonic irradiation has a very strong mixing and dispersion effect, much stronger than conventional stirring, which can improve the dispersion of titania/cellulose-SO 3 H nanoparticles in the polymer matrix. The strong -SO 3 H/-SO 3 H interaction between s-PBI chains and titania/cellulose-SO 3 H hybrids leads to ionic cross-linking in the membrane structure, which increases both the thermal stability and methanol resistance of the membranes. After acid doping with phosphoric acid, s-PBI/titania/cellulose-SO 3 H nanocomposite membranes exhibit depressions on methanol permeability and enhancements on proton conductivity comparing to the pristine s-PBI membrane. The chemical structure of the functionlized titania was characterized with FTIR, and energy-dispersive X-ray. Imidazole and sulfonated groups on the surface of modified nanoparticles forming linkages with s-PBI chains, improved the compatibility between s-PBI and nanoparticles, and enhanced the mechanical strength of the prepared nanocomposite membranes. From SEM and TEM analysis could explain the homogeneous dispersion of titania/cellulose-SO 3 H in nanocomposite membranes. Moreover, the membranes exhibited excellent antibacterial activities against S. aureus and E. coli. A. Copyright © 2017 Elsevier B.V. All rights reserved.

  20. Development of Novel active transport membrane devices. Phase I. Final report, 31 October 1988--31 January 1994

    Energy Technology Data Exchange (ETDEWEB)

    Laciak, D.V.; Quinn, R.; Choe, G.S.; Cook, P.J.; Tsai, Fu-Jya

    1994-08-01

    The main objective of this program was to identify and develop a technique for fabricating Active Transport Materials (ATM) into lab-scale membrane devices. Air Products met this objective by applying thin film, multilayer fabrication techniques to support the AT material on a substrate membrane. In Phase IA, spiral-wound hollow fiber membrane modules were fabricated and evaluated. These nonoptimized devices were used to demonstrate the AT-based separation of carbon dioxide from methane, hydrogen sulfide from methane, and ammonia from hydrogen. It was determined that a need exists for a more cost efficient and less energy intensive process for upgrading subquality natural gas. Air Products estimated the effectiveness of ATM for this application and concluded that an optimized ATM system could compete effectively with both conventional acid gas scrubbing technology and current membrane technology. In addition, the optimized ATM system would have lower methane loss and consume less energy than current alternative processes. Air Products made significant progress toward the ultimate goal of commercializing an advanced membrane for upgrading subquality natural gas. The laboratory program focused on developing a high performance hollow fiber substrate and fabricating and evaluating ATM-coated lab-scale hollow fiber membrane modules. Selection criteria for hollow fiber composite membrane supports were developed and used to evaluate candidate polymer compositions. A poly(amide-imide), PAI, was identified for further study. Conditions were identified which produced microporous PAI support membrane with tunable surface porosity in the range 100-1000{Angstrom}. The support fibers exhibited good hydrocarbon resistance and acceptable tensile strength though a higher elongation may ultimately be desirable. ATM materials were coated onto commercial and PAI substrate fiber. Modules containing 1-50 fibers were evaluated for permselectivity, pressure stability, and lifetime.

  1. The Role of Long-Term Tectonic Deformation on the Distribution of Present-Day Seismic Activity in the Caribbean and Central America

    Science.gov (United States)

    Schobelock, J.; Stamps, D. S.; Pagani, M.; Garcia, J.; Styron, R. H.

    2017-12-01

    The Caribbean and Central America region (CCAR) undergoes the entire spectrum of earthquake types due to its complex tectonic setting comprised of transform zones, young oceanic spreading ridges, and subductions along its eastern and western boundaries. CCAR is, therefore, an ideal setting in which to study the impacts of long-term tectonic deformation on the distribution of present-day seismic activity. In this work, we develop a continuous tectonic strain rate model based on inter-seismic geodetic data and compare it with known active faults and earthquake focal mechanism data. We first create a 0.25o x 0.25o finite element mesh that is comprised of block geometries defined in previously studies. Second, we isolate and remove transient signals from the latest open access community velocity solution from UNAVCO, which includes 339 velocities from COCONet and TLALOCNet GNSS data for the Caribbean and Central America, respectively. In a third step we define zones of deformation and rigidity by creating a buffer around the boundary of each block that varies depending on the size of the block and the expected deformation zone based on locations of GNSS data that are consistent with rigid block motion. We then assign each node within the buffer a 0 for the deforming areas and a plate index outside the buffer for the rigid. Finally, we calculate a tectonic strain rate model for CCAR using the Haines and Holt finite element approach to fit bi-cubic Bessel splines to the the GNSS/GPS data assuming block rotation for zones of rigidity. Our model of the CCAR is consistent with compression along subduction zones, extension across the mid-Pacific Rise, and a combination of compression and extension across the North America - Caribbean plate boundary. The majority of CCAR strain rate magnitudes range from -60 to 60 nanostrains/yr. Modeling results are then used to calculate expected faulting behaviors that we compare with mapped geologic faults and seismic activity.

  2. Activation of the B cell receptor leads to increased membrane proximity of the Igα cytoplasmic domain.

    Directory of Open Access Journals (Sweden)

    Wing-Yiu Lee

    Full Text Available Binding of antigen to the B cell receptor (BCR induces conformational changes in BCR's cytoplasmic domains that are concomitant with phosphorylation of the immunoreceptor tyrosine-based activation motifs (ITAMs. Recently, reversible folding of the CD3ε and ξ chain ITAMs into the plasma membrane has been suggested to regulate T cell receptor signaling. Here we show that the Igα and Igβ cytoplasmic domains of the BCR do not associate with plasma membrane in resting B cells. However, antigen binding and ITAM phosphorylation specifically increased membrane proximity of Igα, but not Igβ. Thus, BCR activation is accompanied by asymmetric conformational changes, possibly promoting the binding of Igα and Igβ to differently localized signaling complexes.

  3. Evolution of fault activity reflecting the crustal deformation: Insights from crustal stress and fault orientations in the northeast-southwest Japan

    Science.gov (United States)

    Miyakawa, A.; Otsubo, M.

    2015-12-01

    We evaluated fault activity in northeast- southwest Japan based on the regional stress and the fault orientation field for both active faults and inactive faults (here, an inactive fault is a fault which activity has not been identified in Quaternary). The regional stress field was calculated using the stress inversion method [Hardebeck and Michael, 2006] applied to earthquake focal mechanisms in the northeast-southwest Japan. The locations and orientations (i.e., strike and dip, assuming a planar fault geometry) of active faults in the study area were obtained from the Active Fault Database of Japan and inactive faults from a database compiled by Kosaka et al. [2011]. We employed slip tendency analysis [Morris et al., 1996] to evaluate the likelihood of fault slip. The values of the slip tendency is generally higher along active faults than along inactive faults. The difference between the slip tendencies of active and inactive faults reflects the difference in their activities. Furthermore the high slip tendency observed for some inactive faults suggests their high activity. These high slip tendencies imply that they have potential to be active. We propose the temporal evolution from inactive to active faulting during long-term crustal deformation to explain the potential for fault activity along inactive faults. When a region undergoes the transition from inactive to active faulting, potential active faults are observed as inactive faults with a high Part of this findig have been submitted to Tectonics (AGU Journal) (2015-07-27). We will presentate some new findings.slip tendency. The average slip tendency of inactive faults gradually increases from northeast to southwest Japan, because a relatively large number of inactive faults in southwest Japan have a high slip tendency. The representative deformation zones in Japan shows a relationship with the observed spatial variations in the evolution from inactive to active faulting. This study was supported by MEXT

  4. Quantification of functional groups and modeling of their ionization behavior in the active layer of FT30 reverse osmosis membrane.

    Science.gov (United States)

    Coronell, Orlando; Mariñas, Benito J; Zhang, Xijing; Cahill, David G

    2008-07-15

    A new experimental approach was developed to measure the concentration of charged functional groups (FGs) in the active layer of thin-film composite reverse osmosis (RO) and nanofiltration (NF) membranes as a function of solution pH. FT30 RO membrane, with a fully aromatic polyamide (PA) active layer sandwiched between a polysulfone support and a coating layer, was used. The experiments consisted of saturating charged FGs with heavy ion probes, and determining the ion probe concentration by Rutherford backscattering spectrometry (RBS). Deprotonated carboxylic groups were saturated with Ag+, and protonated amine groups with W04(2-). The ionization behavior of carboxylic and amine groups was modeled based on acid-base equilibrium theory. While the ionization behavior of amine groups was satisfactorily described by one dissociation constant (pKa = 4.74), two pKa values (5.23 and 8.97) were necessary to describe the titration curve of carboxylic groups. These results were consistent with the bimodal pore size distribution (PSD) of FT30 active layer reported in the literature. The calculated total concentrations of carboxylic and amine groups in the active layer of the FT30 RO membrane studied were 0.432 and 0.036 M, respectively, and the isoelectric point (IEP) was 4.7. The total concentration of carboxylic and amine groups revealed that the degree of cross-linking of the PA active layer of the FT30 RO membrane studied was 94%.

  5. Ca(2+)-activated anion channels and membrane depolarizations induced by blue light and cold in Arabidopsis seedlings

    Science.gov (United States)

    Lewis, B. D.; Karlin-Neumann, G.; Davis, R. W.; Spalding, E. P.; Evans, M. L. (Principal Investigator)

    1997-01-01

    The activation of an anion channel in the plasma membrane of Arabidopsis thaliana hypocotyls by blue light (BL) is believed to be a signal-transducing event leading to growth inhibition. Here we report that the open probability of this particular anion channel depends on cytoplasmic Ca2+ ([Ca2+]cyt) within the concentration range of 1 to 10 microM, raising the possibility that BL activates the anion channel by increasing [Ca2+]cyt. Arabidopsis seedlings cytoplasmically expressing aequorin were generated to test this possibility. Aequorin luminescence did not increase during or after BL, providing evidence that Ca2+ does not play a second-messenger role in the activation of anion channels. However, cold shock simultaneously triggered a large increase in [Ca2+]cyt and a 110-mV transient depolarization of the plasma membrane. A blocker of the anion channel, 5-nitro-2-(3-phenylpropylamino)-benzoic acid, blocked 61% of the cold-induced depolarization without affecting the increase in [Ca2+]cyt. These data led us to propose that cold shock opens Ca2+ channels at the plasma membrane, allowing an inward, depolarizing Ca2+ current. The resulting large increase in [Ca2+]cyt activates the anion channel, which further depolarizes the membrane. Although an increase in [Ca2+]cyt may activate anion channels in response to cold, it appears that BL does so via a Ca(2+)-independent pathway.

  6. [Bacterial outer membrane vesicles as nano carriers to study immunological activities].

    Science.gov (United States)

    Qi, Chen; Min, W U; Hongzhen, Bai; Zeling, Guo; Jun, Zhou; Qingqing, Wang; Guping, Tang

    2017-03-25

    Objective: To prepare a nano-carrier based on combining bacterial outer membrane vesicles (OMV) with three block polymer pluronic F127 (PEO 100 -PPO 65 -PEO 100 ) (OMV-F127) and to investigate its immunological activity. Methods: Attenuated salmonella (sal) was cultivated. OMV were separated by centrifugal ultrafiltration or ultrasonication, and OMV-F127 was prepared by mechanical extrudation method. The protein contents and compositions were tested with BCA and SDS-PAGE; the morphology of OMV, F127 and OMV-F127 were observed with FM and TEM; the particle sizes and their zeta potential were determined with DLS. Mouse macrophage RAW246.7 cells were treated with OMV-F127 (50 μg/mL, 100 μg/mL) in vitro, and the concentrations of IL-12, TNF-α and IFN-γ in culture supernatant were measured with ELISA kits. Results: The contents of protein in separated OMV by centrifugal ultrafiltration and ultrasonication were 2.8 mg/mL and 2.7 mg/mL, respectively. SDS-PAGE showed the marker protein OmpF/C in OMV. Under the FM and TEM, ball-like structure of F127 and OMV-F127 was observed. Size analysis revealed that the diameters of OMV, F127 and OMV-F127 were 72±2 nm, 90±3 nm and 92±2 nm, respectively. ELISA tests revealed that OMV-F127 significantly stimulated the secretion of IL-12, TNF-α and IFN-γ in RAW246.7 cells. Conclusion: A nano-carrier based on bacterial outer membrane vesicles has been prepared, which can stimulate the secretion of cytokines and may have immunomodulatory effects.

  7. Membrane-interaction quantitative structure--activity relationship (MI-QSAR) analyses of skin penetration enhancers.

    Science.gov (United States)

    Zheng, Tao; Hopfinger, A J; Esposito, Emilio X; Liu, Jianzhong; Tseng, Yufeng J

    2008-06-01

    Membrane-interaction quantitative structure-activity relationship (MI-QSAR) models for two skin penetration enhancer data sets of 61 and 42 compounds were constructed and compared to QSAR models constructed for the same two data sets using only classic intramolecular QSAR descriptors. These two data sets involve skin penetration enhancement of hydrocortisone and hydrocortisone acetate, and the enhancers are generally similar in structure to lipids and surfactants. A new MI-QSAR descriptor, the difference in the integrated cylindrical distribution functions over the phospholipid monolayer model, in and out of the presence of the skin penetration enhancer, DeltaSigma h(r), was developed. This descriptor is dominant in the optimized MI-QSAR models of both training sets studied and greatly reduces the size and complexity of the MI-QSAR models as compared to those QSAR models developed using the classic intramolecular descriptors. The MI-QSAR models indicate that good penetration enhancers make bigger "holes" in the monolayer and are less aqueous-soluble, so as to preferentially enter the monolayer, than are poor penetration enhancers. The skin penetration enhancer thus alters the structure and organization of the monolayer. This space and time alteration in the structure and dynamics of the membrane monolayer is captured by DeltaSigma h(r) and is simplistically referred to as "holes" in the monolayer. The MI-QSAR models explain 70-80% of the variance in skin penetration enhancement across each of the two training sets and are stable predictive models using accepted diagnostic measures of robustness and predictivity.

  8. Electrospun polymer membrane activated with room temperature ionic liquid: Novel polymer electrolytes for lithium batteries

    Science.gov (United States)

    Cheruvally, Gouri; Kim, Jae-Kwang; Choi, Jae-Won; Ahn, Jou-Hyeon; Shin, Yong-Jo; Manuel, James; Raghavan, Prasanth; Kim, Ki-Won; Ahn, Hyo-Jun; Choi, Doo Seong; Song, Choong Eui

    A new class of polymer electrolytes (PEs) based on an electrospun polymer membrane incorporating a room-temperature ionic liquid (RTIL) has been prepared and evaluated for suitability in lithium cells. The electrospun poly(vinylidene fluoride- co-hexafluoropropylene) P(VdF-HFP) membrane is activated with a 0.5 M solution of LiTFSI in 1-butyl-3-methylimidazolium bis(trifluoromethanesulfonyl)imide (BMITFSI) or a 0.5 M solution of LiBF 4 in 1-butyl-3-methylimidazolium tetrafluoroborate (BMIBF 4). The resulting PEs have an ionic conductivity of 2.3 × 10 -3 S cm -1 at 25 °C and anodic stability at >4.5 V versus Li +/Li, making them suitable for practical applications in lithium cells. A Li/LiFePO 4 cell with a PE based on BMITFSI delivers high discharge capacities when evaluated at 25 °C at the 0.1 C rate (149 mAh g -1) and the 0.5 C rate (132 mAh g -1). A very stable cycle performance is also exhibited at these low current densities. The properties decrease at the higher, 1 C rate, when operated at 25 °C. Nevertheless, improved properties are obtained at a moderately elevated temperature of operation, i.e. 40 °C. This is attributed to enhanced conductivity of the electrolyte and faster reaction kinetics at higher temperatures. At 40 °C, a reversible capacity of 140 mAh g -1 is obtained at the 1 C rate.

  9. Conditional trimerization and lytic activity of HIV-1 gp41 variants containing the membrane-associated segments.

    Science.gov (United States)

    Dai, Zhou; Tao, Yisong; Liu, Nina; Brenowitz, Michael D; Girvin, Mark E; Lai, Jonathan R

    2015-03-03

    Fusion of host and viral membranes is a critical step during infection by membrane-bound viruses. The HIV-1 glycoproteins gp120 (surface subunit) and gp41 (fusion subunit) represent the prototypic system for studying this process; in the prevailing model, the gp41 ectodomain forms a trimeric six-helix bundle that constitutes a critical intermediate and provides the energetic driving force for overcoming barriers associated with membrane fusion. However, most structural studies of gp41 variants have been performed either on ectodomain constructs lacking one or more of the membrane-associated segments (the fusion peptide, FP, the membrane-proximal external region, MPER, and the transmembrane domain, TM) or on variants consisting of these isolated segments alone without the ectodomain. Several recent reports have suggested that the HIV-1 ectodomain, as well as larger construct containing the membrane-bound segments, dissociates from a trimer to a monomer in detergent micelles. Here we compare the properties of a series of gp41 variants to delineate the roles of the ectodomain, FP, and MPER and TM, all in membrane-mimicking environments. We find that these proteins are prone to formation of a monomer in detergent micelles. In one case, we observed exclusive monomer formation at pH 4 but conditional trimerization at pH 7 even at low micromolar (∼5 μM) protein concentrations. Liposome release assays demonstrate that these gp41-related proteins have the capacity to induce content leakage but that this activity is also strongly modulated by pH with much higher activity at pH 4. Circular dichroism, nuclear magnetic resonance, and binding assays with antibodies specific to the MPER provide insight into the structural and functional roles of the FP, MPER, and TM and their effect on structure within the larger context of the fusion subunit.

  10. Insecticidal Activity Associated with the Outer Membrane Vesicles of Xenorhabdus nematophilus

    OpenAIRE

    Khandelwal, Puneet; Banerjee-Bhatnagar, Nirupama

    2003-01-01

    Xenorhabdus nematophilus secretes a large number of proteins into the culture supernatant as soluble proteins and also as large molecular complexes associated with the outer membrane. Transmission electron micrographs of X. nematophilus cells showed that there was blebbing of the outer membrane from the surface of the bacterium. The naturally secreted outer membrane vesicles (OMVs) were purified from the culture supernatant of X. nematophilus and analyzed. Electron microscopy revealed a vesic...

  11. Activation of PVDF membranes through facile hydroxylation of the polymeric dope

    KAUST Repository

    Al-Gharabli, Samer

    2017-10-30

    A method comprising a two-step alkali/acid treatment of poly (vinylidene fluoride) (PVDF) polymer is developed for the fabrication of flat-sheet PVDF membranes functionalized with labile hydroxyl groups. This method involves the application of a short-duration modification in alkali medium (5% KOH). Extensive characterizations were performed on the prepared membranes. Modification of the polymer altered the crystallinity of the PVDF from a mixture of both α and β phases to a predominant β phase. Lower work of adhesion of the modified membrane indicated the formation of a more hydrophobic and wetting-resistant membrane surface. Centrifugation of the polymer dope after the modification had a pronounced impact on the properties of the resultant membranes. This protocol could be utilized in fine-tuning the properties of PVDF membranes for various target-specific applications such as membrane distillation. This method can also be used in functionalizing PVDF membranes further by exploiting the labile –OH group present on the membrane surface.

  12. Holocene soft-sediment deformation of the Santa Fe-Sopetrán Basin, northern Colombian Andes: Evidence for pre-Hispanic seismic activity?

    Science.gov (United States)

    Suter, F.; Martínez, J. I.; Vélez, M. I.

    2011-04-01

    The detailed study of four deformed intervals from the Holocene fluvio-lacustrine deposits of the Santa Fe-Sopetrán Basin in northern Colombia shows 17 types of soft-sediment deformation (SSD) structures. Evidence indicates that seismic activity was responsible for the SSD structures, a conclusion reached after considering the environmental conditions at the time of sediment deposition and shortly after, and the detailed analysis of the driving force systems. Other triggers (i.e. overloading and rapid sedimentation), however, are not discarded. Intervals showing SSD structures occurred at centennial frequencies and apparently resulted from Mw 6-7 earthquakes. The Holocene age of these major shaking events should be seriously considered when evaluating the seismic hazard and risk for the middle Cauca Valley and the nearby city of Medellín with 3 million inhabitants.

  13. Magnetically controlled permeability membranes

    KAUST Repository

    Kosel, Jurgen

    2013-10-31

    A bioactive material delivery system can include a thermoresponsive polymer membrane and nanowires distributed within the thermoresponsive polymer membrane. Magnetic activation of a thermoresponsive polymer membrane can take place via altering the magnetization or dimensions of nanowires dispersed or ordered within the membrane matrix.

  14. Active Pedicle Epithelial Flap Transposition Combined with Amniotic Membrane Transplantation for Treatment of Nonhealing Corneal Ulcers

    Directory of Open Access Journals (Sweden)

    Ting Zhang

    2016-01-01

    Full Text Available Introduction. The objective was to evaluate the efficacy of active pedicle epithelial flap transposition combined with amniotic membrane transplantation (AMT in treating nonhealing corneal ulcers. Material and Methods. Eleven patients (11 eyes with nonhealing corneal ulcer who underwent the combined surgery were included. Postoperatively, ulcer healing time was detected by corneal fluorescein staining. Visual acuity, intraocular pressure, surgical complications, and recurrence were recorded. Corneal status was inspected by the laser scanning confocal microscopy and anterior segment optical coherence tomography (AS-OCT. Results. The primary diseases were herpes simplex keratitis (8 eyes, corneal graft ulcer (2 eyes, and Stevens-Johnson syndrome (1 eye. All epithelial flaps were intact following surgery, without shedding or displacement. Mean ulcer healing time was 10.8±3.1 days, with a healing rate of 91%. Vision significantly improved from 1.70 to 0.82 log MAR (P=0.001. A significant decrease in inflammatory cell infiltration and corneal stromal edema was revealed 2 months postoperatively by confocal microscopy and AS-OCT. Corneal ulcer recurred in 1 eye. None of the patients developed major complications. Conclusion. Active pedicle epithelial flap transposition combined with AMT is a simple and effective treatment for nonhealing corneal ulcers.

  15. Antifungal Activity of (+-2,2’-Epicytoskyrin A and Its Membrane-Disruptive Action

    Directory of Open Access Journals (Sweden)

    Dewi Wulansari

    2016-12-01

    Full Text Available (+-2,2’-Epicytoskyrin A, a bis-anthraquinone isolated from fungal endophyte Diaporthe sp. GNBP-10 associated with Uncaria gambir Roxb., was investigated for its antifungal activity. The broth microdilution method was used to determine the minimum inhibitory concentration (MIC against 22 yeast strains and three filamentous fungi. The MICs of (+-2,2’-epicytoskyrin A ranged from 16 to 128 µg/mL, which exhibited lower activity than the antifungal nystatin. A study of the mechanism of action revealed similar effects of (+-2,2’-epicytoskyrin A and nystatin on Candida tropicalis at their MICs (16 and 8 µg/mL, respectively and 2 times of the MIC. Both compounds caused cytoplasmic material and ion leakages on fungal cell, which were characterized by an increase in absorbance at 260 nm and 280 nm as well as Ca2+ and K+ ion concentrations. The morphology of the fungal cells after (+-2,2’-epicytoskyrin A treatment was observed under a scanning electron microscope. The control cells, which were not treated with either (+-2,2’-epicytoskyrin A or nystatin, showed a smooth surface, while the cells treated with either (+-2,2’-epicytoskyrin A or nystatin shrank and displayed a donut-like shape. More shrinkage was observed in the 2 times MIC concentration and even more in the cells exposed to nystatin. The action of (+-2,2’-epicytoskyrin A was proposed through membrane disruption.

  16. Active Pedicle Epithelial Flap Transposition Combined with Amniotic Membrane Transplantation for Treatment of Nonhealing Corneal Ulcers.

    Science.gov (United States)

    Zhang, Ting; Wang, Yuexin; Jia, Yanni; Liu, Dongle; Li, Suxia; Shi, Weiyun; Gao, Hua

    2016-01-01

    Introduction . The objective was to evaluate the efficacy of active pedicle epithelial flap transposition combined with amniotic membrane transplantation (AMT) in treating nonhealing corneal ulcers. Material and Methods . Eleven patients (11 eyes) with nonhealing corneal ulcer who underwent the combined surgery were included. Postoperatively, ulcer healing time was detected by corneal fluorescein staining. Visual acuity, intraocular pressure, surgical complications, and recurrence were recorded. Corneal status was inspected by the laser scanning confocal microscopy and anterior segment optical coherence tomography (AS-OCT). Results . The primary diseases were herpes simplex keratitis (8 eyes), corneal graft ulcer (2 eyes), and Stevens-Johnson syndrome (1 eye). All epithelial flaps were intact following surgery, without shedding or displacement. Mean ulcer healing time was 10.8 ± 3.1 days, with a healing rate of 91%. Vision significantly improved from 1.70 to 0.82 log MAR ( P = 0.001). A significant decrease in inflammatory cell infiltration and corneal stromal edema was revealed 2 months postoperatively by confocal microscopy and AS-OCT. Corneal ulcer recurred in 1 eye. None of the patients developed major complications. Conclusion . Active pedicle epithelial flap transposition combined with AMT is a simple and effective treatment for nonhealing corneal ulcers.

  17. Bacterial membrane activity of a-peptide/b-peptoid chimeras: Influence of amino acid composition and chain length on the activity against different bacterial strains

    DEFF Research Database (Denmark)

    Hein-Kristensen, Line; Knapp, Kolja M; Franzyk, Henrik

    2011-01-01

    acid only had a minor effect on MIC values, whereas chain length had a profound influence on activity. All chimeras were less active against Serratia marcescens (MICs above 46 μM). The chimeras were bactericidal and induced leakage of ATP from Staphylococcus aureus and S. marcescens with similar time......BACKGROUND: Characterization and use of antimicrobial peptides (AMPs) requires that their mode of action is determined. The interaction of membrane-active peptides with their target is often established using model membranes, however, the actual permeabilization of live bacterial cells...... and subsequent killing is usually not tested. In this report, six α-peptide/β-peptoid chimeras were examined for the effect of amino acid/peptoid substitutions and chain length on the membrane perturbation and subsequent killing of food-borne and clinical bacterial isolates. RESULTS: All six AMP analogues...

  18. Using fluorescence-activated flow cytometry to determine reactive oxygen species formation and membrane lipid peroxidation in viable boar spermatozoa

    Science.gov (United States)

    Fluorescence-activated flow cytometry analyses were developed for determination of reactive oxygen species (ROS) formation and membrane lipid peroxidation in live spermatozoa loaded with, respectively, hydroethidine (HE) or the lipophilic probe 4,4-difluoro-5-(4-phenyl-1,3-butadienyl)-4-bora-3a,4a-d...

  19. Na+/K(+)pump activity in photoreceptors of the blowfly Calliphora : A model analysis based on membrane potential measurements

    NARCIS (Netherlands)

    Gerster, U; Stavenga, DG; Backhaus, W

    Na+/K+-pump activity and intracellular Na+ and K+ concentration changes in blowfly photoreceptors are derived from intracellular potential measurements in vivo with a model based on the Goldman-Hodgkin-Katz theory for membrane currents. The relation between the intracellular Na+ concentration and

  20. Impact of coagulant and flocculant addition to an anaerobic dynamic membrane bioreactor (AnDMBR) treating waste-activated sludge

    NARCIS (Netherlands)

    Kooijman, G.; Lopes, Wilton; Zhou, Z.; Guo, H.; de Kreuk, M.K.; Spanjers, H.L.F.M.; van Lier, J.B.

    2017-01-01

    In this work, we investigated the effects of flocculation aid (FA) addition to an anaerobic dynamic membrane bioreactor (AnDMBR) (7 L, 35°C) treating waste-activated sludge (WAS). The experiment consisted of three distinct periods. In period 1 (day 1–86), the reactor was operated as a

  1. Imaging active faulting in a region of distributed deformation from the joint clustering of focal mechanisms and hypocentres: Application to the Azores-western Mediterranean region

    Science.gov (United States)

    Custódio, Susana; Lima, Vânia; Vales, Dina; Cesca, Simone; Carrilho, Fernando

    2016-04-01

    The matching between linear trends of hypocentres and fault planes indicated by focal mechanisms (FMs) is frequently used to infer the location and geometry of active faults. This practice works well in regions of fast lithospheric deformation, where earthquake patterns are clear and major structures accommodate the bulk of deformation, but typically fails in regions of slow and distributed deformation. We present a new joint FM and hypocentre cluster algorithm that is able to detect systematically the consistency between hypocentre lineations and FMs, even in regions of distributed deformation. We apply the method to the Azores-western Mediterranean region, with particular emphasis on western Iberia. The analysis relies on a compilation of hypocentres and FMs taken from regional and global earthquake catalogues, academic theses and technical reports, complemented by new FMs for western Iberia. The joint clustering algorithm images both well-known and new seismo-tectonic features. The Azores triple junction is characterised by FMs with vertical pressure (P) axes, in good agreement with the divergent setting, and the Iberian domain is characterised by NW-SE oriented P axes, indicating a response of the lithosphere to the ongoing oblique convergence between Nubia and Eurasia. Several earthquakes remain unclustered in the western Mediterranean domain, which may indicate a response to local stresses. The major regions of consistent faulting that we identify are the mid-Atlantic ridge, the Terceira rift, the Trans-Alboran shear zone and the north coast of Algeria. In addition, other smaller earthquake clusters present a good match between epicentre lineations and FM fault planes. These clusters may signal single active faults or wide zones of distributed but consistent faulting. Mainland Portugal is dominated by strike-slip earthquakes with fault planes coincident with the predominant NNE-SSW and WNW-ESE oriented earthquake lineations. Clusters offshore SW Iberia are

  2. Sphingolipids activate membrane fusion of Semliki Forest virus in a stereospecific manner

    DEFF Research Database (Denmark)

    Moesby, Lise; Corver, J; Erukulla, R K

    1995-01-01

    The alphavirus Semliki Forest virus (SFV) enters cells through receptor-mediated endocytosis. Subsequently, triggered by the acid pH in endosomes, the viral envelope fuses with the endosomal membrane. Membrane fusion of SFV has been shown previously to be dependent on the presence of cholesterol ...

  3. Differences between Solution and Membrane Forms of Chitosan on the In Vitro Activity of Fibroblasts

    Directory of Open Access Journals (Sweden)

    Bahar Uslu

    2015-03-01

    Full Text Available Background: Chitosan, a linear polysaccharide, has been recently used in biomedical applications. In vitro studies have demonstrated its effect on cellular growth and its stimulatory action on cellular layer formation. Aims: The present study aims to compare the proliferative effects of chitosan in two forms, membranous and solution forms, on Swiss 3T3 mouse embryonic fibroblasts. Study Design: In vitro study. Methods: Three experimental groups were formed: cells were cultured in a normal medium without chitosan (Control Group; cells were cultured either in a medium containing 2.0% chitosan in membranous form (Membrane Group or chitosan solution at a concentration of 2.0% (Solution Group.Two different methods were used in the experiments: cells cultured on the medium containing chitosan in solution or membranous forms (method 1; and chitosan solution or membranous forms were added into the medium containing previously cultured cells (method 2. Results: Scanning electron microscopic investigations of the experimental groups revealed cells with well-defined cellular projections, intact cellular membranes and tight intercellular junctions. They were especially prominent in the membrane group of method 1 and in the membrane and solution groups of method 2. Mouse monoclonal anti-collagen 1 primary antibody was used to indicate collagen synthesis. Prominent collagen synthesis was detected in the membrane groups on the 10th day of culture for both methods. Bromodeoxyuridine (BrdU and MTT assays were performed in order to assess cellular proliferation and viability, respectively. BrdU labelling tests indicated a higher proliferation index in the membrane group of method 1 on the 5th and 10th days. For the second method, the membranous form on the 10th day and solution form on the 5th day were the most effective groups in terms of cellular proliferation. MTT results reflected a high cellular viability in method 1 on the 5th day of treatment with the

  4. Effects of Iron Overload on the Activity of Na,K-ATPase and Lipid Profile of the Human Erythrocyte Membrane.

    Directory of Open Access Journals (Sweden)

    Leilismara Sousa

    Full Text Available Iron is an essential chemical element for human life. However, in some pathological conditions, such as hereditary hemochromatosis type 1 (HH1, iron overload induces the production of reactive oxygen species that may lead to lipid peroxidation and a change in the plasma-membrane lipid profile. In this study, we investigated whether iron overload interferes with the Na,K-ATPase activity of the plasma membrane by studying erythrocytes that were obtained from the whole blood of patients suffering from iron overload. Additionally, we treated erythrocytes of normal subjects with 0.8 mM H2O2 and 1 μM FeCl3 for 24 h. We then analyzed the lipid profile, lipid peroxidation and Na,K-ATPase activity of plasma membranes derived from these cells. Iron overload was more frequent in men (87.5% than in women and was associated with an increase (446% in lipid peroxidation, as indicated by the amount of the thiobarbituric acid reactive substances (TBARS and an increase (327% in the Na,K-ATPase activity in the plasma membrane of erythrocytes. Erythrocytes treated with 1 μM FeCl3 for 24 h showed an increase (132% in the Na,K-ATPase activity but no change in the TBARS levels. Iron treatment also decreased the cholesterol and phospholipid content of the erythrocyte membranes and similar decreases were observed in iron overload patients. In contrast, erythrocytes treated with 0.8 mM H2O2 for 24 h showed no change in the measured parameters. These results indicate that erythrocytes from patients with iron overload exhibit higher Na,K-ATPase activity compared with normal subjects and that this effect is specifically associated with altered iron levels.

  5. A role for phospholipase A2 activity in membrane tubule formation and TGN trafficking.

    Science.gov (United States)

    Schmidt, John A; Kalkofen, Danielle N; Donovan, Kirk W; Brown, William J

    2010-12-01

    We have investigated the role of phospholipase A(2) (PLA(2) ) enzymes in generating membrane tubules at the trans-Golgi network (TGN). Constitutive TGN membrane tubules and those induced by over-expressing kinase dead protein kinase D were inhibited by the PLA(2) inhibitors ONO-RS-082 (ONO) and bromoenol lactone. These antagonists also inhibited secretory delivery of both soluble and transmembrane cargoes. Finally, use of the reversible antagonist ONO and time-lapse imaging revealed for the first time that PLA(2) antagonists inhibit the initiation of membrane tubule formation at the TGN. Thus, PLA(2) enzymes appear to have an important role in the earliest steps of membrane tubule formation at the TGN, which are utilized for membrane trafficking. © 2010 John Wiley & Sons A/S.

  6. Modulation of membrane potential by an acetylcholine-activated potassium current in trout atrial myocytes

    DEFF Research Database (Denmark)

    Molina, C.E.; Gesser, Hans; Llach, A.

    2007-01-01

    Application of the current-clamp technique in rainbow trout atrial myocytes has yielded resting membrane potentials that are incompatible with normal atrial function. To investigate this paradox, we recorded the whole membrane current (Im) and compared membrane potentials recorded in isolated...... cardiac myocytes and multicellular preparations. Atrial tissue and ventricular myocytes had stable resting potentials of -87 ± 2 mV and -83.9 ± 0.4 mV, respectively. In contrast, 50 out of 59 atrial myocytes had unstable depolarized membrane potentials that were sensitive to the holding current. We...... hypothesized that this is at least partly due to a small slope conductance of Im around the resting membrane potential in atrial myocytes. In accordance with this hypothesis, the slope conductance of Im was about sevenfold smaller in atrial than in ventricular myocytes. Interestingly, ACh increased Im at -120...

  7. Nonlinear electro-mechanobiological behavior of cell membrane during electroporation

    KAUST Repository

    Deng, Peigang

    2012-01-01

    A nonlinear electroporation (EP) model is proposed to study the electro-mechanobiological behavior of cell membrane during EP, by taking the nonlinear large deformation of the membrane into account. The proposed model predicts the critical transmembrane potential and the activation energy for EP, the equilibrium pore size, and the resealing process of the pore. Single-cell EP experiments using a micro EP chip were conducted on chicken red blood cells at different temperatures to determine the activation energy and the critical transmembrane potential for EP. The experimental results are in good agreement with the theoretical predictions. © 2012 American Institute of Physics.

  8. Preferential inhibition of the plasma membrane NADH oxidase (NOX) activity by diphenyleneiodonium chloride with NADPH as donor

    Science.gov (United States)

    Morre, D. James

    2002-01-01

    The cell-surface NADH oxidase (NOX) protein of plant and animal cells will utilize both NADH and NADPH as reduced electron donors for activity. The two activities are distinguished by a differential inhibition by the redox inhibitor diphenyleneiodonium chloride (DPI). Using both plasma membranes and cells, activity with NADPH as donor was markedly inhibited by DPI at submicromolar concentrations, whereas with NADH as donor, DPI was much less effective or had no effect on the activity. The possibility of the inhibition being the result of two different enzymes was eliminated by the use of a recombinant NOX protein. The findings support the concept that NOX proteins serve as terminal oxidases for plasma membrane electron transport involving cytosolic reduced pyridine nucleotides as the natural electron donors and with molecular oxygen as the electron acceptor.

  9. Contribution of the complement Membrane Attack Complex to the bactericidal activity of human serum.

    Science.gov (United States)

    Berends, Evelien T M; Mohan, Sarbani; Miellet, Willem R; Ruyken, Maartje; Rooijakkers, Suzan H M

    2015-06-01

    Direct killing of Gram-negative bacteria by serum is usually attributed to the Membrane Attack Complex (MAC) that is assembled upon activation of the complement system. In serum bactericidal assays, the activity of the MAC is usually blocked by a relatively unspecific method in which certain heat-labile complement components are inactivated at 56°C. The goal of this study was to re-evaluate MAC-driven lysis towards various Gram-negative bacteria. Instead of using heat-treatment, we included the highly specific C5 cleavage inhibitor OmCI to specifically block the formation of the MAC. Using a C5 conversion analysis tool, we monitored the efficacy of the inhibitor during the incubations. Our findings indicate that 'serum-sensitive' bacteria are not necessarily killed by the MAC. Other heat-labile serum factors can contribute to serum bactericidal activity. These unidentified factors are most potent at serum concentrations of 10% and higher. Furthermore, we also find that some bacteria can be killed by the MAC at a slower rate. Our data demonstrate the requirement for the use of specific inhibitors in serum bactericidal assays and revealed that the classification of serum-sensitive and resistant strains needs re-evaluation. Moreover, it is important to determine bacterial viability at multiple time intervals to differentiate serum susceptibility between bacterial species. In conclusion, these data provide new insights into bacterial killing by the humoral immune system and may guide future vaccine development studies for the treatment of pathogenic serum-resistant bacteria. Copyright © 2015 Elsevier Ltd. All rights reserved.

  10. Calibrating a side-stream membrane bioreactor using Activated Sludge Model No. 1.

    Science.gov (United States)

    Jiang, T; Liu, X; Kennedy, M D; Schippers, J C; Vanrolleghem, P A

    2005-01-01

    Membrane bioreactors (MBRs) are attracting global interest but the mathematical modeling of the biological performance of MBRs remains very limited. This study focuses on the modeling of a side-stream MBR system using the Activated Sludge Model No. 1 (ASM1), and compares the results with the modeling of traditional activated sludge processes. ASM1 parameters relevant for the long-term biological behaviour in MBR systems were calibrated (i.e. Y(H) = 0.72 gCOD/gCOD, Y(A) = 0.25 gCOD/gN, b(H) = 0.25 d(-1), b(A) = 0.080 d(-1) and f(p) = 0.06), and generally agreed with the parameters in traditional activated sludge processes, with the exception that a higher autotrophic biomass decay rate was observed in the MBR. Influent wastewater characterization was proven to be a critical step in model calibration, and special care should be taken in characterizing the inert particulate COD (X(I)) concentration in the MBR influent. It appeared that the chemical-biological method was superior to the physical-chemical method. A sensitivity analysis for steady-state operation and DO dynamics suggested that the biological performance of the MBR system (the sludge concentration, effluent quality and the DO dynamics) are very sensitive to the parameters (i.e. Y(H), Y(A), b(H), b(A) micro(maxH) and micro(maxA), and influent wastewater components (X(I), S(s), X(s) and S(NH)).

  11. Depression of membrane-bound Na+-K+-ATPase activity induced by free radicals and by ischemia of kidney

    International Nuclear Information System (INIS)

    Kako, K.; Kato, M.; Matsuoka, T.; Mustapha, A.

    1988-01-01

    A partially purified, membrane-bound Na + -K + -ATPase fraction, prepared from the outer medulla of porcine kidney, was incubated in the presence of 0.1-100 mM H 2 O 2 for either 15 or 30 min at 37 degree C. The activity of ouabain-sensitive Na + -K + -ATPase was reduced proportionally to the concentration of H 2 O 2 and the duration of incubation. There were decreases in SH contents and turnover rates of the Na + -K + -ATPase preparation, while malondialdehyde (MDA) and conjugated dienes were generated from the membrane lipids in the course of the incubation. The concentrations of ethanolamine (E) plasmalogen and of arachidonic acid in the E glycerophospholipid molecules were reduced by the free radical reaction. Similarly, a reduction in Na + K + -ATPase activity and the formation of MDA and conjugated dienes, together with a decrease in E glycerophospholipids, were observed when the membrane fraction was exposed to ultraviolet irradiation (254 nm) for 30 min at 4 degree C. Microsomal fractions, prepared from the outer medulla of canine kidney after 1 h of unilateral ischemia and 1 h of reperfusion, showed a decreased Na + -K + -ATPase activity, a reduced amount of SH groups, and an increased MDA. These changes were normalized by the infusion of N-mercaptopropionylglycine. These results support the view (1) that free radical generation affects the enzyme protein as well as membrane lipids, and (2) that free radicals may be formed in the ischemic reperfused kidney

  12. Ubiquitination of the bacterial inositol phosphatase, SopB, regulates its biological activity at the plasma membrane.

    LENUS (Irish Health Repository)

    Knodler, Leigh A

    2009-11-01

    The Salmonella type III effector, SopB, is an inositol polyphosphate phosphatase that modulates host cell phospholipids at the plasma membrane and the nascent Salmonella-containing vacuole (SCV). Translocated SopB persists for many hours after infection and is ubiquitinated but the significance of this covalent modification has not been investigated. Here we identify by mass spectrometry six lysine residues of SopB that are mono-ubiquitinated. Substitution of these six lysine residues with arginine, SopB-K(6)R, almost completely eliminated SopB ubiquitination. We found that ubiquitination does not affect SopB stability or membrane association, or SopB-dependent events in SCV biogenesis. However, two spatially and temporally distinct events are dependent on ubiquitination, downregulation of SopB activity at the plasma membrane and prolonged retention of SopB on the SCV. Activation of the mammalian pro-survival kinase Akt\\/PKB, a downstream target of SopB, was intensified and prolonged after infection with the SopB-K(6)R mutant. At later times, fewer SCV were decorated with SopB-K(6)R compared with SopB. Instead SopB-K(6)R was present as discrete vesicles spread diffusely throughout the cell. Altogether, our data show that ubiquitination of SopB is not related to its intracellular stability but rather regulates its enzymatic activity at the plasma membrane and intracellular localization.

  13. Thrombus imaging in a primate model with antibodies specific for an external membrane protein of activated platelets

    International Nuclear Information System (INIS)

    Palabrica, T.M.; Furie, B.C.; Konstam, M.A.; Aronovitz, M.J.; Connolly, R.; Brockway, B.A.; Ramberg, K.L.; Furie, B.

    1989-01-01

    The activated platelet is a potential target for the localization of thrombi in vivo since, after stimulation and secretion of granule contents, activated platelets are concentrated at sites of blood clot formation. In this study, we used antibodies specific for a membrane protein of activated platelets to detect experimental thrombi in an animal model. PADGEM (platelet activation-dependent granule-external membrane protein), a platelet alpha-granule membrane protein, is translocated to the plasma membrane during platelet activation and granule secretion. Since PADGEM is internal in unstimulated platelets, polyclonal anti-PADGEM and monoclonal KC4 antibodies do not bind to circulating resting platelets but do interact with activated platelets. Dacron graft material incubated with radiolabeled KC4 or anti-PADGEM antibodies in the presence of thrombin-activated platelet-rich plasma bound most of the antibody. Imaging experiments with 123I-labeled anti-PADGEM in baboons with an external arterial-venous Dacron shunt revealed rapid uptake in the thrombus induced by the Dacron graft; control experiments with 123I-labeled nonimmune IgG exhibited minimal uptake. Deep venous thrombi, formed by using percutaneous balloon catheters to stop blood flow in the femoral vein of baboons, were visualized with 123I-labeled anti-PADGEM. Thrombi were discernible against blood pool background activity without subtraction techniques within 1 hr. No target enhancement was seen with 123I-labeled nonimmune IgG. 123I-labeled anti-PADGEM cleared the blood pool with an initial half-disappearance time of 6 min and did not interfere with hemostasis. These results indicate that radioimmunoscintigraphy with anti-PADGEM antibodies can visualize thrombi in baboon models and is a promising technique for clinical thrombus detection in humans

  14. ASM1-based activated sludge model with biopolymer kinetics for integrated simulation of membrane bioreactors for wastewater treatment

    OpenAIRE

    Janus, Tomasz; Ulanicki, Bogumil

    2015-01-01

    This paper presents an activated sludge model suitable for modelling membrane bioreactors (MBRs) for wastewater treatment. The model, later referred to as combined EPS and SMP production ASM1-based model (CES-ASM1), extends Activated Sludge Model No. 1 (ASM1) with biokinetics of two types of bacterial biopolymers: soluble microbial products (SMP) and extracellular polymeric substances (EPS). The biopolymer kinetics in CES-ASM1 are, in their majority, borrowed from Laspidou and Rittmann[1] ...

  15. Specific chlamydial inclusion membrane proteins associate with active Src family kinases in microdomains that interact with the host microtubule network.

    Science.gov (United States)

    Mital, Jeffrey; Miller, Natalie J; Fischer, Elizabeth R; Hackstadt, Ted

    2010-09-01

    Chlamydiae are Gram-negative obligate intracellular bacteria that cause diseases with significant medical and economic impact. Chlamydia trachomatis replicates within a vacuole termed an inclusion, which is extensively modified by the insertion of a number of bacterial effector proteins known as inclusion membrane proteins (Incs). Once modified, the inclusion is trafficked in a dynein-dependent manner to the microtubule-organizing centre (MTOC), where it associates with host centrosomes. Here we describe a novel structure on the inclusion membrane comprised of both host and bacterial proteins. Members of the Src family of kinases are recruited to the chlamydial inclusion in an active form. These kinases display a distinct, localized punctate microdomain-like staining pattern on the inclusion membrane that colocalizes with four chlamydial inclusion membrane proteins (Incs) and is enriched in cholesterol. Biochemical studies show that at least two of these Incs stably interact with one another. Furthermore, host centrosomes associate with these microdomain proteins in C. trachomatis-infected cells and in uninfected cells exogenously expressing one of the chlamydial effectors. Together, the data suggest that a specific structure on the C. trachomatis inclusion membrane may be responsible for the known interactions of chlamydiae with the microtubule network and resultant effects on centrosome stability.

  16. Effect of powdered activated carbon dosage on sludge properties and membrane bioreactor performance in a hybrid MBR-PAC system.

    Science.gov (United States)

    Zhang, Shi; Zuo, Xingtao; Xiong, Juan; Ma, Cong; Hu, Bo

    2017-12-22

    An improved insight into the effect of powdered activated carbon (PAC) on membrane fouling is crucial to the MBR performance. Sludge key property, soluble microbial products (SMP) and transmembrane pressure (TMP) were monitored. The membrane fouling rate in the MBRs was also analyzed based on TMP profile and resistance-in-series model. PAC reduced the membrane filtration resistance and significantly decreased the fouling rate. The sludge filterability was improved by extending the filtration time by almost twofold. PAC affected the SMP release and protein/polysaccharide (carbohydrate) was in a lower ratio. Fourier transform infrared (FTIR) analysis indicated that PAC decreased the impact of organic carbon, and reduced the proteins' and polysaccharides' absorption and deposition on the membrane surface and in the pores. The degree of reversible and irreversible fouling was related to the PAC content added into the MBRs. At the optimum dosage of 2 g/L, the results signified the PAC potential as a mitigation strategy of membrane fouling.

  17. Monocyte activation drives preservation of membrane thiols by promoting release of oxidised membrane moieties via extracellular vesicles.

    Science.gov (United States)

    Szabó-Taylor, K É; Tóth, E Á; Balogh, A M; Sódar, B W; Kádár, L; Pálóczi, K; Fekete, N; Németh, A; Osteikoetxea, X; Vukman, K V; Holub, M; Pállinger, É; Nagy, Gy; Winyard, P G; Buzás, E I

    2017-07-01

    The redox state of cellular exofacial molecules is reflected by the amount of available thiols. Furthermore, surface thiols can be considered as indicators of immune cell activation. One group of thiol containing proteins, peroxiredoxins, in particular, have been associated with inflammation. In this study, we assessed surface thiols of the U937 and Thp1 monocyte cell lines and primary monocytes in vitro upon inflammatory stimulation by irreversibly labelling the cells with a fluorescent derivative of maleimide. We also investigated exofacial thiols on circulating blood mononuclear cells in patients with rheumatoid arthritis and healthy controls. When analysing extracellular vesicles, we combined thiol labelling with the use of antibodies to specific CD markers to exclude extracellular vesicle mimicking signals from thiol containing protein aggregates. Furthermore, differential detergent lysis was applied to confirm the vesicular nature of the detected extracellular events in blood plasma. We found an increase in exofacial thiols on monocytes upon in vitro stimulation by LPS or TNF, both in primary monocytes and monocytic cell lines (pextracellular vesicles showed a decrease in their exofacial thiols compared with those from unstimulated cells (pextracellular vesicles of isolated CD14 + cells from rheumatoid arthritis patients had decreased thiol levels compared with healthy subjects (pextracellular vesicles was increased in rheumatoid arthritis blood plasma (pextracellular vesicle-enriched preparations from blood plasma. Our data show that cell surface thiols play a protective role and reflect oxidative stress resistance state in activated immune cells. Furthermore, they support a role of extracellular vesicles in the redox regulation of human monocytes, possibly representing an antioxidant mechanism. Copyright © 2017. Published by Elsevier Inc.

  18. The extracellular membrane-proximal domain of membrane-bound IgE restricts B cell activation by limiting B cell antigen receptor surface expression.

    Science.gov (United States)

    Vanshylla, Kanika; Opazo, Felipe; Gronke, Konrad; Wienands, Jürgen; Engels, Niklas

    2018-03-01

    Immunoglobulin E (IgE) antibodies are key mediators of allergic reactions. Due to their potentially harmful anaphylactic properties, their production is tightly regulated. The membrane-bound isoform of IgE (mIgE), which is an integral component of the B cell antigen receptor, has been shown to be critical for the regulation of IgE responses in mice. In primate species including humans, mIgE can be expressed in two isoforms that are produced by alternative splicing of the primary ε Ig heavy chain transcript, and differ in the absence or presence of an extracellular membrane-proximal domain (EMPD) consisting of 52 amino acids. However, the function of the EMPD remains unclear. Here, we demonstrate that the EMPD restricts surface expression of mIgE-containing BCRs in human and murine B cells. The EMPD does not interfere with BCR assembly but acts as an autonomous endoplasmic reticulum retention domain. Limited surface expression of EMPD-containing mIgE-BCRs caused impaired activation of intracellular signaling cascades and hence represents a regulatory mechanism that may control the production of potentially anaphylactic IgE antibodies in primate species. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  19. Photodynamic Action of LED-Activated Curcumin against Staphylococcus aureus Involving Intracellular ROS Increase and Membrane Damage

    Directory of Open Access Journals (Sweden)

    Yuan Jiang

    2014-01-01

    Full Text Available Aim. To investigate the effect of photodynamic action of LED-activated curcumin on cell viability, membrane permeability, and intracellular reactive oxygen species of Staphylococcus aureus. Methods. Staphylococcus aureus was incubated with the different concentrations of curcumin for 60 min and then irradiated by blue light with the wavelength of 470 nm and with light dose of 3 J/cm2. The colony forming unit assay was used to investigate photocytotoxicity of curcumin on Staphylococcus aureus, confocal laser scanning microscopy (CLSM and flow cytometry (FCM for assaying membrane permeability, FCM analysis with DCFH-DA staining for measuring the intracellular ROS level, and transmission electron microscopy (TEM for observing morphology and structure. Results. Blue light-activated curcumin significantly killed Staphylococcus aureus in a curcumin dose-dependent manner. TEM observed remarkable structural damages in S. aureus after light-activated curcumin. More red fluorescence of PI dye was found in S. aureus treated by blue light-activated curcumin than in those of the controlled bacterial cells. Intracellular ROS increase was observed after light-activated curcumin. Conclusion. Blue light-activated curcumin markedly damaged membrane permeability, resulting in cell death of Staphylococcus aureus and highlighted that intracellular ROS increase might be an important event in photodynamic killing of Staphylococcus aureus in the presence of curcumin.

  20. Removal of emerging contaminants by simultaneous application of membrane ultrafiltration, activated carbon adsorption, and ultrasound irradiation

    Energy Technology Data Exchange (ETDEWEB)

    Secondes, Mona Freda N. [Environmental Engineering Graduate Program, Department of Chemical Engineering, University of the Philippines – Diliman, Quezon City (Philippines); Naddeo, Vincenzo, E-mail: vnaddeo@unisa.it [Sanitary and Environmental Engineering Division (SEED), Department of Civil Engineering, University of Salerno, Fisciano 84084 (Saudi Arabia) (Italy); Belgiorno, Vincenzo [Sanitary and Environmental Engineering Division (SEED), Department of Civil Engineering, University of Salerno, Fisciano 84084 (Saudi Arabia) (Italy); Ballesteros, Florencio [Environmental Engineering Graduate Program, Department of Chemical Engineering, University of the Philippines – Diliman, Quezon City (Philippines)

    2014-01-15

    Highlights: • Above 99% of the emerging contaminants were removed in the USAMe process. • Influence of PAC dose and US frequency on removal is studied. • Improved performance is due to PAC adsorption enhancement and sonolytic degradation. • US irradiation improved efficiency and delayed declines in the removal of contaminants. • Performance of the hybrid process is better under lower frequency ultrasound irradiation. -- Abstract: Advanced wastewater treatment is necessary to effectively remove emerging contaminants (ECs) with chronic toxicity, endocrine disrupting effects, and the capability to induce the proliferation of highly resistant microbial strains in the environment from before wastewater disposal or reuse. This paper investigates the efficiency of a novel hybrid process that applies membrane ultrafiltration, activated carbon adsorption, and ultrasound irradiation simultaneously to remove ECs. Diclofenac, carbamazepine, and amoxicillin are chosen for this investigation because of their assessed significant environmental risks. Removal mechanisms and enhancement effects are analysed in single and combined processes. The influence of adsorbent dose and ultrasonic frequency to EC removal are also investigated. Results suggest that adsorption is probably the main removal mechanism and is affected by the nature of ECs and the presence of other components in the mixture. Almost complete removals are achieved in the hybrid process for all ECs.

  1. Removal of emerging contaminants by simultaneous application of membrane ultrafiltration, activated carbon adsorption, and ultrasound irradiation

    International Nuclear Information System (INIS)

    Secondes, Mona Freda N.; Naddeo, Vincenzo; Belgiorno, Vincenzo; Ballesteros, Florencio

    2014-01-01

    Highlights: • Above 99% of the emerging contaminants were removed in the USAMe process. • Influence of PAC dose and US frequency on removal is studied. • Improved performance is due to PAC adsorption enhancement and sonolytic degradation. • US irradiation improved efficiency and delayed declines in the removal of contaminants. • Performance of the hybrid process is better under lower frequency ultrasound irradiation. -- Abstract: Advanced wastewater treatment is necessary to effectively remove emerging contaminants (ECs) with chronic toxicity, endocrine disrupting effects, and the capability to induce the proliferation of highly resistant microbial strains in the environment from before wastewater disposal or reuse. This paper investigates the efficiency of a novel hybrid process that applies membrane ultrafiltration, activated carbon adsorption, and ultrasound irradiation simultaneously to remove ECs. Diclofenac, carbamazepine, and amoxicillin are chosen for this investigation because of their assessed significant environmental risks. Removal mechanisms and enhancement effects are analysed in single and combined processes. The influence of adsorbent dose and ultrasonic frequency to EC removal are also investigated. Results suggest that adsorption is probably the main removal mechanism and is affected by the nature of ECs and the presence of other components in the mixture. Almost complete removals are achieved in the hybrid process for all ECs

  2. Performance enhancement with powdered activated carbon (PAC) addition in a membrane bioreactor (MBR) treating distillery effluent

    International Nuclear Information System (INIS)

    Satyawali, Yamini; Balakrishnan, Malini

    2009-01-01

    This work investigated the effect of powdered activated carbon (PAC) addition on the operation of a membrane bioreactor (MBR) treating sugarcane molasses based distillery wastewater (spentwash). The 8 L reactor was equipped with a submerged 30 μm nylon mesh filter with 0.05 m 2 filtration area. Detailed characterization of the commercial wood charcoal based PAC was performed before using it in the MBR. The MBR was operated over 200 days at organic loading rates (OLRs) varying from 4.2 to 6.9 kg m -3 d -1 . PAC addition controlled the reactor foaming during start up and enhanced the critical flux by around 23%; it also prolonged the duration between filter cleaning. Operation at higher loading rates was possible and for a given OLR, the chemical oxygen demand (COD) removal was higher with PAC addition. However, biodegradation in the reactor was limited and the high molecular weight compounds were not affected by PAC supplementation. The functional groups on PAC appear to interact with the polysaccharide portion of the sludge, which may reduce its propensity to interact with the nylon mesh.

  3. Membrane Fouling Potential of Secondary Effluent Organic Matter (EfOM) from Conventional Activated Sludge Process

    KAUST Repository

    Wei, Chunhai

    2012-01-01

    Secondary effluent organic matter (EfOM) from a conventional activated sludge process was filtered through constant-pressure dead-end filtration tests with a sequential ultrafiltration (UF, molecular weight cut-off (MWCO) of 10k Dalton) and nanofiltration (NF, MWCO of 200 Dalton) array to investigate its membrane fouling potential. Advanced analytical methods including liquid chromatography with online carbon detection (LC-OCD) and fluorescent excitation-emission matrix (F-EEM) were employed for EfOM characterization. EfOM consisted of humic substances and building blocks, low molecular weight (LMW) neutrals, biopolymers (mainly proteins) and hydrophobic organics according to the sequence of their organic carbon fractions. The UF rejected only biopolymers and the NF rejected most humics and building blocks and a significant part of LMW neutrals. Simultaneous occurrence of cake layer and standard blocking during the filtration process of both UF and NF was identified according to constant-pressure filtration equations, which was possibly caused by the heterogeneous nature of EfOM with a wide MW distribution (several ten to several million Dalton). Thus the corresponding two fouling indices (kc for cake layer and ks for standard blocking) from UF and NF could characterize the fouling potential of macromolecular biopolymers and low to intermediate MW organics (including humics, building blocks, LMW neutrals), respectively. Compared with macromolecular biopolymers, low to intermediate MW organics exhibited a much higher fouling potential due to their lower molecular weight and higher concentration.

  4. Assessment of activated sludge, membrane bioreactors and vertical flow wetlands for upgrading sewage treatment works.

    Science.gov (United States)

    Besançon, A; Le Corre, K S; Dotro, G; Jefferson, B

    2017-01-01

    This paper demonstrates that utilising a vertical flow (VF) wetland after a conventional activated sludge (CAS) delivers equivalent or better effluent quality to a membrane bioreactor (MBR) based on a side-by-side pilot trial. The CAS was operated under the solids retention times (SRT) of 6, 12, and 20 days, with the effluent from each pilot plant fed onto a soil aquifer treatment column to better understand their water reuse application potential. Results showed an upgraded CAS + VF system could deliver effluents with median values of 34 mgO 2 .L -1 , 7 mg.L -1 and 1.9 mg.L -1 for organics, solids and ammonia nitrogen, respectively, which were statistically similar to those from the MBR. Water reuse standards were achieved by the upgraded system for most parameters, with the exception of total coliform removal. The upgraded system delivered superior metal removal when compared to the CAS. An economic analysis showed upgrading a CAS with a VF wetland was more favourable than investing in an MBR system for example works of 5000 and 50,000 population equivalents if the VF system was operated at hydraulic loading rates of 0.03 m.d -1 and 0.08 m.d -1 , respectively. This was delivered for a tenth of the carbon footprint of the MBR treatment.

  5. Restoration of active transport of solutes and oxidative phosphorylation by naphthoquinones in irradiated membrane vesicles from Mycobacterium phlei

    Science.gov (United States)

    Lee, Soon-Ho; Sutherland, Thomas O.; Deves, Rosa; Brodie, Arnold F.

    1980-01-01

    Irradiation of the inverted membrane vesicles of Mycobacterium phlei with light at 360 nm inactivated the natural menaquinone [MK9(II-H)] and resulted in a loss of substrate oxidation, pH gradient, membrane potential, active transport of proline or calcium ions, and oxidative phosphorylation. Restoration of the protonmotive force and active transport occurred on addition of naphthoquinones such as vitamin K1, menadione, or lapachol to the irradiated membrane vesicles. However, coupled phosphorylation was restored only by vitamin K1. Menadione and lapachol did not act as uncoupling agents. The magnitude of the pH gradient and membrane potential in the quinone-restored system was a reflection of the rate of oxidation and was correlated with the rate of uptake of proline or Ca2+. These results are consistent with the chemosmotic hypothesis proposed for the energy transducing mechanism for active transport and further demonstrate that the complete respiratory chain is not required to drive active transport. In contrast, the data suggest that in addition to the driving force (protonmotive force) necessary to establish oxidative phosphorylation, a specific spatial orientation of the respiratory components, such as the naphthaquinones, is essential for the utilization of the proton gradient or membrane potential or both. Bypass of electrons from the respiratory chain with menadione may explain the inability of this quinone to restore oxidative phosphorylation; however, lapachol restores oxidation by the same electron transport pathway as the natural menaquinone but fails to restore phosphorylation. Because all three quinones restore the protonmotive force, other factors that are discussed must be considered in understanding the mechanism of oxidative phosphorylation. PMID:6928606

  6. Spectrins: a structural platform for stabilization and activation of membrane channels, receptors and transporters.

    Science.gov (United States)

    Machnicka, Beata; Czogalla, Aleksander; Hryniewicz-Jankowska, Anita; Bogusławska, Dżamila M; Grochowalska, Renata; Heger, Elżbieta; Sikorski, Aleksander F

    2014-02-01

    This review focuses on structure and functions of spectrin as a major component of the membrane skeleton. Recent advances on spectrin function as an interface for signal transduction mediation and a number of data concerning interaction of spectrin with membrane channels, adhesion molecules, receptors and transporters draw a picture of multifaceted protein. Here, we attempted to show the current depiction of multitask role of spectrin in cell physiology. This article is part of a Special Issue entitled: Reciprocal influences between cell cytoskeleton and membrane channels, receptors and transporters. Guest Editor: Jean Claude Hervé. Copyright © 2013 Elsevier B.V. All rights reserved.

  7. [Effect of damage integrity rat brain synaptic membranes on the functional activity GABA(A)-receptor/Cl(-)-ionophore complex in the CNC].

    Science.gov (United States)

    Rebrov, I G; Kalinina, M V

    2013-01-01

    Functional activity of the CGABA(A)-receptor/Cl(-) ionophore complex was investigated the muscimol-stimulated entry of the radioactive isotope 36Cl(-) in synaptoneurosomes in changing the structure and permeability of neuronal membranes. Integrity of the membranes was damaged by removal of Ca(+2) and Mg(+2) from the incubation medium and by the method of freezing-thawing synaptoneurosomes. In both cases, an increase in basal 36Cl(-) entry into synaptoneurosomes, indicating increased nonspecific permeability of neuronal membranes, and decreased activity the CABA(A)-receptor/Cl(-) ionophore complex. The conclusion about the relationship of processes damage neuronal membranes and reducing the inhibitory processes in the epileptic focus.

  8. Peptides actively transported across the tympanic membrane: Functional and structural properties.

    Directory of Open Access Journals (Sweden)

    Arwa Kurabi

    Full Text Available Otitis media (OM is the most common infectious disease of children under six, causing more antibiotic prescriptions and surgical procedures than any other pediatric condition. By screening a bacteriophage (phage library genetically engineered to express random peptides on their surfaces, we discovered unique peptides that actively transport phage particles across the intact tympanic membrane (TM and into the middle ear (ME. Herein our goals were to characterize the physiochemical peptide features that may underlie trans-TM phage transport; assess morphological and functional effects of phage peptides on the ME and inner ear (IE; and determine whether peptide-bearing phage transmigrate from the ME into the IE. Incubation of five peptide-bearing phage on the TM for over 4hrs resulted in demonstrably superior transport of one peptide, in level and in exponential increase over time. This suggests a preferred peptide motif for TM active transport. Functional and structural comparisons revealed unique features of this peptide: These include a central lysine residue, isoelectric point of 0.0 at physiological pH and a hydrophobic C-terminus. When the optimal peptide was applied to the TM independent of phage, similar transport was observed, indicating that integration into phage is not required. When 109 particles of the four different trans-TM phage were applied directly into the ME, no morphological effects were detected in the ME or IE when compared to saline or wild-type (WT phage controls. Comparable, reversible hearing loss was observed for saline controls, WT phage and trans-TM peptide phage, suggesting a mild conductive hearing loss due to ME fluid. Perilymph titers after ME incubation established that few copies of trans-TM peptide phage crossed into the IE. The results suggest that, within the parameters tested, trans-TM peptides are safe and could be used as potential agents for noninvasive delivery of drugs, particles and gene therapy

  9. Static response of deformable microchannels

    Science.gov (United States)

    Christov, Ivan C.; Sidhore, Tanmay C.

    2017-11-01

    Microfluidic channels manufactured from PDMS are a key component of lab-on-a-chip devices. Experimentally, rectangular microchannels are found to deform into a non-rectangular cross-section due to fluid-structure interactions. Deformation affects the flow profile, which results in a nonlinear relationship between the volumetric flow rate and the pressure drop. We develop a framework, within the lubrication approximation (l >> w >> h), to self-consistently derive flow rate-pressure drop relations. Emphasis is placed on handling different types of elastic response: from pure plate-bending, to half-space deformation, to membrane stretching. The ``simplest'' model (Stokes flow in a 3D rectangular channel capped with a linearly elastic Kirchhoff-Love plate) agrees well with recent experiments. We also simulate the static response of such microfluidic channels under laminar flow conditions using ANSYSWorkbench. Simulations are calibrated using experimental flow rate-pressure drop data from the literature. The simulations provide highly resolved deformation profiles, which are difficult to measure experimentally. By comparing simulations, experiments and our theoretical models, we show good agreement in many flow/deformation regimes, without any fitting parameters.

  10. In vivo characterization of the scaffold activity of flotillin on the membrane kinase KinC of Bacillus subtilis

    Science.gov (United States)

    Schneider, Johannes; Mielich-Süss, Benjamin; Böhme, Richard; Lopez, Daniel

    2015-01-01

    Summary Scaffold proteins are ubiquitous chaperones that bind to proteins and facilitate the physical interaction of the components of signal transduction pathways or multi-enzymatic complexes. In this study, we used a biochemical approach to dissect the molecular mechanism of a membrane-associated scaffold protein, FloT, a flotillin-homolog protein that is localized in Functional Membrane Microdomains of the bacterium Bacillus subtilis. This study provides unambiguous evidence that FloT physically binds to and interacts with the membrane-bound sensor kinase KinC. This sensor kinase activates biofilm formation in B. subtilis in response to the presence of the self-produced signal surfactin. Furthermore, we have characterized the mechanism by which the interaction of FloT with KinC benefits the activity of KinC. Two separate and synergistic effects constitute this mechanism: first, the scaffold activity of FloT promotes more efficient self-interaction of KinC and facilitates dimerization into its active form. Second, the selective binding of FloT to KinC prevents the occurrence of unspecific aggregation between KinC and other proteins that may generate dead-end intermediates that could titrate the activity of KinC. Flotillin proteins appear to play an important role in prokaryotes in promoting effective binding of signaling proteins with their correct protein partners. PMID:26297017

  11. Fluorescent mannosides serve as acceptor substrates for glycosyltransferase and sugar-1-phosphate transferase activities in Euglena gracilis membranes.

    Science.gov (United States)

    Ivanova, Irina M; Nepogodiev, Sergey A; Saalbach, Gerhard; O'Neill, Ellis C; Urbaniak, Michael D; Ferguson, Michael A J; Gurcha, Sudagar S; Besra, Gurdyal S; Field, Robert A

    2017-01-13

    Synthetic hexynyl α-D-mannopyranoside and its α-1,6-linked disaccharide counterpart were fluorescently labelled through CuAAC click chemistry with 3-azido-7-hydroxycoumarin. The resulting triazolyl-coumarin adducts, which were amenable to analysis by TLC, HPLC and mass spectrometry, proved to be acceptor substrates for α-1,6-ManT activities in mycobacterial membranes, as well as α- and β-GalT activities in trypanosomal membranes, benchmarking the potential of the fluorescent acceptor approach against earlier radiochemical assays. Following on to explore the glycobiology of the benign protozoan alga Euglena gracilis, α-1,3- and α-1,2-ManT activities were detected in membrane preparations, along with GlcT, Glc-P-T and GlcNAc-P-T activities. These studies serve to demonstrate the potential of readily accessible fluorescent glycans as substrates for exploring carbohydrate active enzymes. Copyright © 2016 The Authors. Published by Elsevier Ltd.. All rights reserved.

  12. A Ca-activated K channel from rabbit renal brush-border membrane vesicles in planar lipid bilayers.

    Science.gov (United States)

    Zweifach, A; Desir, G V; Aronson, P S; Giebisch, G H

    1991-07-01

    Rabbit renal brush-border membranes were fused to planar lipid bilayers to gain insight into the nature and properties of ion channels from the luminal membrane of the proximal tubule. Fusion was obtained using osmotic gradients. A large conductance channel was commonly observed. Measurements of reversal potentials indicated that the channel was selective for K over Rb, Na, and Cl. Channel open probability was increased by membrane depolarization and by increased Ca activity on the intracellular face of the channel. The channel was inhibited by charybdotoxin (CTX), a protein from leiurus venom, from the external side of the channel. The channel was also blocked by Ba and quinidine added to the intracellular bathing solution. Na added to the intracellular bathing solution reduced current amplitude in a voltage-dependent fashion. In addition, methylisobutyl amiloride, an analogue of the K-sparing diuretic amiloride, inhibited channel activity when added to the external solution. The possible physiological role of the channel is discussed. The usefulness to the study of renal ion channels of the technique of fusing membrane vesicles to planar lipid bilayers is evaluated.

  13. Roles of specific membrane lipid domains in EGF receptor activation and cell adhesion molecule stabilization in a developing olfactory system.

    Science.gov (United States)

    Gibson, Nicholas J; Tolbert, Leslie P; Oland, Lynne A

    2009-09-29

    Reciprocal interactions between glial cells and olfactory receptor neurons (ORNs) cause ORN axons entering the brain to sort, to fasciculate into bundles destined for specific glomeruli, and to form stable protoglomeruli in the developing olfactory system of an experimentally advantageous animal species, the moth Manduca sexta. Epidermal growth factor receptors (EGFRs) and the cell adhesion molecules (IgCAMs) neuroglian and fasciclin II are known to be important players in these processes. We report in situ and cell-culture studies that suggest a role for glycosphingolipid-rich membrane subdomains in neuron-glia interactions. Disruption of these subdomains by the use of methyl-beta-cyclodextrin results in loss of EGFR activation, depletion of fasciclin II in ORN axons, and loss of neuroglian stabilization in the membrane. At the cellular level, disruption leads to aberrant ORN axon trajectories, small antennal lobes, abnormal arrays of olfactory glomerul, and loss of normal glial cell migration. We propose that glycosphingolipid-rich membrane subdomains (possible membrane rafts or platforms) are essential for IgCAM-mediated EGFR activation and for anchoring of neuroglian to the cytoskeleton, both required for normal extension and sorting of ORN axons.

  14. Membrane phospholipid fatty acid composition regulates cardiac SERCA activity in a hibernator, the Syrian hamster (Mesocricetus auratus.

    Directory of Open Access Journals (Sweden)

    Sylvain Giroud

    Full Text Available Polyunsaturated fatty acids (PUFA have strong effects on hibernation and daily torpor. Increased dietary uptake of PUFA of the n-6 class, particularly of Linoleic acid (LA, C18:2 n-6 lengthens torpor bout duration and enables animals to reach lower body temperatures (T(b and metabolic rates. As previously hypothesized, this well-known influence of PUFA may be mediated via effects of the membrane fatty acid composition on sarcoplasmic reticulum (SR Ca(2+-ATPase 2a (SERCA in the heart of hibernators. We tested the hypotheses that high proportions of n-6 PUFA in general, or specifically high proportions of LA (C18:2 n-6 in SR phospholipids (PL should be associated with increased cardiac SERCA activity, and should allow animals to reach lower minimum T(b in torpor. We measured activity of SERCA from hearts of hibernating and non-hibernating Syrian hamsters (Mesocricetus auratus in vitro at 35 °C. Further, we determined the PL fatty acid composition of the SR membrane of these hearts. We found that SERCA activity strongly increased as the proportion of LA in SR PL increased but was negatively affected by the content of Docosahexaenoic acid (DHA; C22:6 n-3. SR PL from hibernating hamsters were characterized by high proportions of LA and low proportions of DHA. As a result, SERCA activity was significantly higher during entrance into torpor and in torpor compared to inter-bout arousal. Also, animals with increased SERCA activity reached lower T(b during torpor. Interestingly, a subgroup of hamsters which never entered torpor but remained euthermic throughout winter displayed a phenotype similar to animals in summer. This was characterized by lower proportions of LA and increased proportions of DHA in SR membranes, which is apparently incompatible with torpor. We conclude that the PUFA composition of SR membranes affects cardiac function via modulating SERCA activity, and hence determines the minimum T(b tolerated by hibernators.

  15. Activity of tick antimicrobial peptide from Ixodes persulcatus (persulcatusin) against cell membranes of drug-resistant Staphylococcus aureus.

    Science.gov (United States)

    Miyoshi, Naruhide; Isogai, Emiko; Hiramatsu, Keiichi; Sasaki, Takashi

    2017-02-01

    Persulcatusin (IP), which is an antimicrobial peptide found in Ixodes persulcatus midgut, is active against Gram-positive bacteria such as Staphylococcus aureus. Multidrug-resistant bacteria in particular methicillin-resistant S. aureus (MRSA), vancomycin-intermediate S. aureus (VISA) and vancomycin-resistant S. aureus (VRSA) are a worldwide clinical concern. In the present study, to explore the potential of IP as a new agent against multidrug-resistant S. aureus infections, we evaluated the antimicrobial activity of IP against multidrug-resistant S. aureus strains by MIC 90 , morphological observation with scanning electron microscope (SEM), and the calcein leakage assay of membrane integrity. Among the six antimicrobial peptides used in this study, IP exhibited the lowest MIC 90 values for both vancomycin-susceptible and -resistant S. aureus strains. The IP MIC 90 against a VISA strain was equivalent to vancomycin, while the MIC 90 against VRSA was relatively low. SEM observations indicated that bacterial cells exposed to IP were crumpled and showed prominent structural changes. Moreover, IP influenced the cell membranes of both MRSA and VRSA in a mere 5 min, leading to leakage of the preloaded calcein. Although a VISA strain was resistant to the action of IP on cell membrane, the MIC 90 of IP was lower than that of Nisin, suggesting that IP had another bactericidal mechanism in addition to cell membrane attack. Our results indicate that the synthetic tick antimicrobial peptide, IP exhibits strong antibacterial activity against multidrug-resistant S. aureus strains, including VRSA, via both cell membrane attack and another unknown mechanism. IP represents a promising candidate for a new anti-VRSA therapy.

  16. Type-1 cannabinoid receptors reduce membrane fluidity of capacitated boar sperm by impairing their activation by bicarbonate.

    Directory of Open Access Journals (Sweden)

    Barbara Barboni

    Full Text Available BACKGROUND: Mammalian spermatozoa acquire their full fertilizing ability (so called capacitation within the female genital tract, where they are progressively exposed to inverse gradients of inhibiting and stimulating molecules. METHODOLOGY/PRINCIPAL FINDINGS: In the present research, the effect on this process of anandamide, an endocannabinoid that can either activate or inhibit cannabinoid receptors depending on its concentration, and bicarbonate, an oviductal activatory molecule, was assessed, in order to study the role exerted by the type 1 cannabinoid receptor (CB1R in the process of lipid membrane remodeling crucial to complete capacitation. To this aim, boar sperm were incubated in vitro under capacitating conditions (stimulated by bicarbonate in the presence or in the absence of methanandamide (Met-AEA, a non-hydrolysable analogue of anandamide. The CB1R involvement was studied by using the specific inhibitor (SR141716 or mimicking its activation by adding a permeable cAMP analogue (8Br-cAMP. By an immunocytochemistry approach it was shown that the Met-AEA inhibits the bicarbonate-dependent translocation of CB1R from the post-equatorial to equatorial region of sperm head. In addition it was found that Met-AEA is able to prevent the bicarbonate-induced increase in membrane disorder and the cholesterol extraction, both preliminary to capacitation, acting through a CB1R-cAMP mediated pathway, as indicated by MC540 and filipin staining, EPR spectroscopy and biochemical analysis on whole membranes (CB1R activity and on membrane enriched fraction (C/P content and anisotropy. CONCLUSIONS/SIGNIFICANCE: Altogether, these data demonstrate that the endocannabinoid system strongly inhibits the process of sperm capacitation, acting as membrane stabilizing agent, thus increasing the basic knowledge on capacitation-related signaling and potentially opening new perspectives in diagnostics and therapeutics of male infertility.

  17. Meningococcal outer membrane vesicle composition-dependent activation of the innate immune response

    NARCIS (Netherlands)

    Zariri, Afshin; Beskers, Joep; van de Waterbeemd, Bas; Hamstra, Hendrik Jan; Bindels, Tim H E; van Riet, Elly; van Putten, Jos P M; van der Ley, Peter

    2016-01-01

    Meningococcal outer membrane vesicles (OMVs) have been extensively investigated and successfully implemented as vaccines. They contain pathogen associated molecular patterns including lipopolysaccharide (LPS), capable of triggering innate immunity. However, Neisseria meningitidis contains an

  18. Advanced Supported Liquid Membranes for Carbon Dioxide Control in Extravehicular Activity Applications

    Science.gov (United States)

    Wickham, David T. (Inventor); Gleason, Kevin J. (Inventor); Cowley, Scott W. (Inventor)

    2015-01-01

    There is disclosed a portable life support system with a component for removal of at least one selected gas. In an embodiment, the system includes a supported liquid membrane having a first side and a second side in opposition to one another, the first side configured for disposition toward an astronaut and the second side configured for disposition toward a vacuum atmosphere. The system further includes an ionic liquid disposed between the first side and the second side of the supported liquid membrane, the ionic liquid configured for removal of at least one selected gas from a region housing the astronaut adjacent the first side of the supported liquid membrane to the vacuum atmosphere adjacent the second side of the supported liquid membrane. Other embodiments are also disclosed.

  19. Lesser Himalayan sequences in Eastern Himalaya and their deformation: Implications for Paleoproterozoic tectonic activity along the northern margin of India

    Directory of Open Access Journals (Sweden)

    Dilip Saha

    2013-05-01

    Full Text Available Substantial part of the northern margin of Indian plate is subducted beneath the Eurasian plate during the Caenozoic Himalayan orogeny, obscuring older tectonic events in the Lesser Himalaya known to host Proterozoic sedimentary successions and granitic bodies. Tectonostratigraphic units of the Proterozoic Lesser Himalayan sequence (LHS of Eastern Himalaya, namely the Daling Group in Sikkim and the Bomdila Group in Arunachal Pradesh, provide clues to the nature and extent of Proterozoic passive margin sedimentation, their involvement in pre-Himalayan orogeny and implications for supercontinent reconstruction. The Daling Group, consisting of flaggy quartzite, meta-greywacke and metapelite with minor mafic dyke and sill, and the overlying Buxa Formation with stromatolitic carbonate-quartzite-slate, represent shallow marine, passive margin platformal association. Similar lithostratigraphy and broad depositional framework, and available geochronological data from intrusive granites in Eastern Himalaya indicate strikewise continuity of a shallow marine Paleoproterozoic platformal sequence up to Arunachal Pradesh through Bhutan. Multiple fold sets and tectonic foliations in LHS formed during partial or complete closure of the sea/ocean along the northern margin of Paleoproterozoic India. Such deformation fabrics are absent in the upper Palaeozoic–Mesozoic Gondwana formations in the Lesser Himalaya of Darjeeling-Sikkim indicating influence of older orogeny. Kinematic analysis based on microstructure, and garnet composition suggest Paleoproterozoic deformation and metamorphism of LHS to be distinct from those associated with the foreland propagating thrust systems of the Caenozoic Himalayan collisional belt. Two possibilities are argued here: (1 the low greenschist facies domain in the LHS enveloped the amphibolite to granulite facies domains, which were later tectonically severed; (2 the older deformation and metamorphism relate to a Pacific type

  20. Application of Pre-coated Microfiltration Ceramic Membrane with Powdered Activated Carbon for Natural Organic Matter Removal from Secondary Wastewater Effluent

    KAUST Repository

    Kurniasari, Novita

    2012-12-01

    Ceramic membranes offer more advantageous performances than conventional polymeric membranes. However, membrane fouling caused by Natural Organic Matters (NOM) contained in the feed water is still become a major problem for operational efficiency. A new method of ceramic membrane pre-coating with Powdered Activated Carbon (PAC), which allows extremely contact time for adsorbing aquatic contaminants, has been studied as a pre-treatment prior to ceramic microfiltration membrane. This bench scale study evaluated five different types of PAC (SA Super, G 60, KCU 6, KCU 8 and KCU 12,). The results showed that KCU 6 with larger pore size was performed better compared to other PAC when pre-coated on membrane surface. PAC pre-coating on the ceramic membrane with KCU 6 was significantly enhance NOM removal, reduced membrane fouling and improved membrane performance. Increase of total membrane resistance was suppressed to 96%. The removal of NOM components up to 92%, 58% and 56% for biopolymers, humic substances and building blocks, respectively was achieved at pre-coating dose of 30 mg/l. Adsorption was found to be the major removal mechanism of NOM. Results obtained showed that biopolymers removal are potentially correlated with enhanced membrane performance.

  1. ABBA regulates plasma-membrane and actin dynamics to promote radial glia extension.

    Science.gov (United States)

    Saarikangas, Juha; Hakanen, Janne; Mattila, Pieta K; Grumet, Martin; Salminen, Marjo; Lappalainen, Pekka

    2008-05-01

    Radial glia play key roles in neuronal migration, axon guidance, and neurogenesis during development of the central nervous system. However, the molecular mechanisms regulating growth and morphology of these extended cells are unknown. We show that ABBA, a novel member of the IRSp53-MIM protein family, is enriched in different types of radial glia. ABBA binds ATP-actin monomers with high affinity and deforms PtdIns(4,5)P(2)-rich membranes in vitro through its WH2 and IM domains, respectively. In radial-glia-like C6-R cells, ABBA localises to the interface between the actin cytoskeleton and plasma membrane, and its depletion by RNAi led to defects in lamellipodial dynamics and process extension. Together, this study identifies ABBA as a novel regulator of actin and plasma membrane dynamics in radial glial cells, and provides evidence that membrane binding and deformation activity is critical for the cellular functions of IRSp53-MIM-ABBA family proteins.

  2. Effects of wheat germ agglutinin on tunicate egg activation and fertilization: is there a plasma membrane sperm receptor system on Ascidia ceratodes eggs?

    Science.gov (United States)

    Flannery, B; Epel, D

    1998-06-01

    Little work has been carried out on the sperm recognition systems present on the egg plasma membrane. Here it is shown that wheat germ agglutinin (WGA) interferes with the sperm-interacting system on the plasma membrane of eggs of the ascidian, Ascidia ceratodes. The WGA activates the dechorionated egg, indicating that a plasma membrane sugar residue can be directly tied to egg activation. Low concentrations of this lectin do not activate the eggs, but reduce fertilizability. This observation suggests that the WGA binding site might be part of a sperm reception-activation complex in the plasma membrane. While WGA also affects sperm binding to the chorion, the mechanisms of sperm interaction at the plasma membrane and chorion show different sensitivities to lectins, sugars and enzymes.

  3. Herpes simplex viruses activate phospholipid scramblase to redistribute phosphatidylserines and Akt to the outer leaflet of the plasma membrane and promote viral entry

    OpenAIRE

    Cheshenko, Natalia; Pierce, Carl; Herold, Betsy C.

    2018-01-01

    Herpes simplex virus (HSV) entry is associated with Akt translocation to the outer leaflet of the plasma membrane to promote a complex signaling cascade. We hypothesized that phospholipid scramblase-1 (PLSCR1), a calcium responsive enzyme that flips phosphatidylserines between membrane leaflets, might redistribute Akt to the outside during entry. Confocal imaging, biotinylation of membrane proteins and flow cytometric analysis demonstrated that HSV activates PLSCR1 and flips phosphatidylserin...

  4. Constraining fault activity by investigating tectonically-deformed Quaternary palaeoshorelines using a synchronous correlation method: the Capo D'Orlando Fault as a case study (NE Sicily, Italy)

    Science.gov (United States)

    Meschis, Marco; Roberts, Gerald P.; Robertson, Jennifer

    2016-04-01

    Long-term curstal extension rates, accommodated by active normal faults, can be constrained by investigating Late Quaternary vertical movements. Sequences of marine terraces tectonically deformed by active faults mark the interaction between tectonic activity, sea-level changes and active faulting throughout the Quaternary (e.g. Armijo et al., 1996, Giunta et al, 2011, Roberts et al., 2013). Crustal deformation can be calculated over multiple seismic cycles by mapping Quaternary tectonically-deformed palaeoshorelines, both in the hangingwall and footwall of active normal faults (Roberts et al., 2013). Here we use a synchronous correlation method between palaeoshorelines elevations and the ages of sea-level highstands (see Roberts et al., 2013 for further details) which takes advantage of the facts that (i) sea-level highstands are not evenly-spaced in time, yet must correlate with palaeoshorelines that are commonly not evenly-spaced in elevation, and (ii) that older terraces may be destroyed and/or overprinted by younger highstands, so that the next higher or lower paleoshoreline does not necessarily correlate with the next older or younger sea-level highstand. We investigated a flight of Late Quaternary marine terraces deformed by normal faulting as a result of the Capo D'Orlando Fault in NE Sicily (e.g. Giunta et al., 2011). This fault lies within the Calabrian Arc which has experienced damaging seismic events such as the 1908 Messina Straits earthquake ~ Mw 7. Our mapping and previous mapping (Giunta et al. (2011) demonstrate that the elevations of marine terraces inner edges change along the strike the NE - SW oriented normal fault. This confirms active deformation on the Capo D'Orlando Fault, strongly suggesting that it should be added into the Database of Individual Seismogenic Sources (DISS, Basili et al., 2008). Giunta et al. (2011) suggested that uplift rates and hence faults lip-rates vary through time for this examples. We update the ages assigned to

  5. Cadmium induces changes in sucrose partitioning, invertase activities, and membrane functionality in roots of Rangpur lime (Citrus limonia L. Osbeck).

    Science.gov (United States)

    Podazza, G; Rosa, M; González, J A; Hilal, M; Prado, F E

    2006-09-01

    Cadmium (Cd) uptake effects on sucrose content, invertase activities, and plasma membrane functionality were investigated in Rangpur lime roots ( CITRUS LIMONIA L. Osbeck). Cadmium accumulation was significant in roots but not in shoots and leaves. Cadmium produced significant reduction in roots DW and increment in WC. Leaves and shoots did not show significant differences on both parameters. Sucrose content was higher in control roots than in Cd-exposed ones. Apoplastic sucrose content was much higher in Cd-exposed roots than in control ones. Cd-exposed roots showed a significant decrease in both cell wall-bound and cytoplasmic (neutral) invertase activities; while the vacuolar isoform did not show any change. Alterations in lipid composition and membrane fluidity of Cd-exposed roots were also observed. In Cd-exposed roots phospholipid and glycolipid contents decreased about 50 %, while sterols content was reduced about 22 %. Proton extrusion was inhibited by Cd. Lipid peroxidation and proton extrusion inhibition were also detected by histochemical analysis. This work's findings demonstrate that Cd affects sucrose partitioning and invertase activities in apoplastic and symplastic regions in Rangpur lime roots as well as the plasma membrane functionality and H (+)-ATPase activity.

  6. Effect of zinc insertion and hydrophobicity on the membrane interactions and PDT activity of porphyrin photosensitizers.

    Science.gov (United States)

    Pavani, Christiane; Uchoa, Adjaci F; Oliveira, Carla S; Iamamoto, Yassuko; Baptista, Maurício S

    2009-02-01

    A series of photosensitizers (PS), which are meso-substituted tetra-cationic porphyrins, was synthesized in order to study the role of amphiphilicity and zinc insertion in photodynamic therapy (PDT) efficacy. Several properties of the PS were evaluated and compared within the series including photophysical properties (absorption spectra, fluorescence quantum yield Phif, and singlet oxygen quantum yield PhiDelta), uptake by vesicles, mitochondria and HeLa cells, dark and phototoxicity in HeLa cells. The photophysical properties of all compounds are quite similar (Phifmembrane potential, i.e., in uncoupled mitochondria PS binding decreases by up to 53%. The porphyrin substituted with octyl groups (TC8PyP) is the compound that is most enriched in mitochondria, and its zinc derivative (ZnTC8PyP) has the highest global uptake. The stronger membrane interaction of the zinc-substituted porphyrins is attributed to a complexing effect with phosphate groups of the phospholipids. Zinc insertion was also shown to decrease the interaction with isolated mitochondria and with the mitochondria of HeLa cells, an effect that has been explained by the particular characteristics of the mitochondrial internal membrane. Phototoxicity was shown to increase proportionally with membrane binding efficiency, which is attributed to favorable membrane interactions which allow more efficient membrane photooxidation. For this series of compounds, photodynamic efficiency is directly proportional to the membrane binding and cell uptake, but it is not totally related to mitochondrial targeting.

  7. Mitigation of Membrane Biofouling in MBR Using a Cellulolytic Bacterium, Undibacterium sp. DM-1, Isolated from Activated Sludge.

    Science.gov (United States)

    Nahm, Chang Hyun; Lee, Seonki; Lee, Sang Hyun; Lee, Kibaek; Lee, Jaewoo; Kwon, Hyeokpil; Choo, Kwang-Ho; Lee, Jung-Kee; Jang, Jae Young; Lee, Chung-Hak; Park, Pyung-Kyu

    2017-03-28

    Biofilm formation on the membrane surface results in the loss of permeability in membrane bioreactors (MBRs) for wastewater treatment. Studies have revealed that cellulose is not only produced by a number of bacterial species but also plays a key role during formation of their biofilm. Hence, in this study, cellulase was introduced to a MBR as a cellulose-induced biofilm control strategy. For practical application of cellulase to MBR, a cellulolytic ( i.e ., cellulase-producing) bacterium, Undibacterium sp. DM-1, was isolated from a lab-scale MBR for wastewater treatment. Prior to its application to MBR, it was confirmed that the cell-free supernatant of DM-1 was capable of inhibiting biofilm formation and of detaching the mature biofilm of activated sludge and cellulose-producing bacteria. This suggested that cellulase could be an effective anti-biofouling agent for MBRs used in wastewater treatment. Undibacterium sp. DM-1-entrapping beads ( i.e ., cellulolytic-beads) were applied to a continuous MBR to mitigate membrane biofouling 2.2-fold, compared with an MBR with vacant-beads as a control. Subsequent analysis of the cellulose content in the biofilm formed on the membrane surface revealed that this mitigation was associated with an approximately 30% reduction in cellulose by cellulolytic-beads in MBR.

  8. Angiotensin II-induced hypertension increases plasma membrane Na pump activity by enhancing Na entry in rat thick ascending limbs.

    Science.gov (United States)

    Gonzalez-Vicente, Agustin; Garvin, Jeffrey L

    2013-11-01

    Thick ascending limbs (TAL) reabsorb 30% of the filtered NaCl load. Na enters the cells via apical Na-K-2Cl cotransporters and Na/H exchangers and exits via basolateral Na pumps. Chronic angiotensin II (ANG II) infusion increases net TAL Na transport and Na apical entry; however, little is known about its effects on the basolateral Na pump. We hypothesized that in rat TALs Na pump activity is enhanced by ANG II-infusion, a model of ANG II-induced hypertension. Rats were infused with 200 ng·kg(-1)·min(-1) ANG II or vehicle for 7 days, and TAL suspensions were obtained. We studied plasma membrane Na pump activity by measuring changes in 1) intracellular Na (Nai) induced by ouabain; and 2) ouabain-sensitive oxygen consumption (QO2). We found that the ouabain-sensitive rise in Nai in TALs from ANG II-infused rats was 12.8 ± 0.4 arbitrary fluorescent units (AFU)·mg(-1)·min(-1) compared with only 9.9 ± 1.1 AFU·mg(-1)·min(-1) in controls (P Na pump expression, the number of Na pumps in the plasma membrane, or the affinity for Na. When furosemide (1.1 mg·kg(-1)·day(-1)) was coinfused with ANG II, no increase in plasma membrane Na pump activity was observed. We concluded that in ANG II-induced hypertension Na pump activity is increased in the plasma membrane of TALs and that this increase is caused by the chronically enhanced Na entry occurring in this model.

  9. High Specific Selectivity and Membrane-Active Mechanism of Synthetic Cationic Hybrid Antimicrobial Peptides Based on the Peptide FV7.

    Science.gov (United States)

    Tan, Tingting; Wu, Di; Li, Weizhong; Zheng, Xin; Li, Weifen; Shan, Anshan

    2017-02-06

    Hybrid peptides integrating different functional domains of peptides have many advantages, such as remarkable antimicrobial activity, lower hemolysis and ideal cell selectivity, compared with natural antimicrobial peptides. FV7 (FRIRVRV-NH₂), a consensus amphiphilic sequence was identified as being analogous to host defense peptides. In this study, we designed a series of hybrid peptides FV7-LL-37 (17-29) (FV-LL), FV7-magainin 2 (9-21) (FV-MA) and FV7-cecropin A (1-8) (FV-CE) by combining the FV7 sequence with the small functional sequences LL-37 (17-29) (LL), magainin 2 (9-21) (MA) and cecropin A (1-8) (CE) which all come from well-described natural peptides. The results demonstrated that the synthetic hybrid peptides, in particular FV-LL, had potent antibacterial activities over a wide range of Gram-negative and Gram-positive bacteria with lower hemolytic activity than other peptides. Furthermore, fluorescent spectroscopy indicated that the hybrid peptide FV-LL exhibited marked membrane destruction by inducing outer and inner bacterial membrane permeabilization, while scanning electron microscopy (SEM) and transmission electron microscopy (TEM) demonstrated that FV-LL damaged membrane integrity by disrupting the bacterial membrane. Inhibiting biofilm formation assays also showed that FV-LL had similar anti-biofilm activity compared with the functional peptide sequence FV7. Synthetic cationic hybrid peptides based on FV7 could provide new models for combining different functional domains and demonstrate effective avenues to screen for novel antimicrobial agents.

  10. Optical waveguide lightmode spectroscopic techniques for investigating membrane-bound ion channel activities.

    Directory of Open Access Journals (Sweden)

    Inna Székács

    Full Text Available Optical waveguide lightmode spectroscopic (OWLS techniques were probed for monitoring ion permeation through channels incorporated into artificial lipid environment. A novel sensor set-up was developed by depositing liposomes or cell-derived membrane fragments onto hydrophilic polytetrafluoroethylene (PTFE membrane. The fibrous material of PTFE membrane could entrap lipoid vesicles and the water-filled pores provided environment for the hydrophilic domains of lipid-embedded proteins. The sensor surface was kept clean from the lipid holder PTFE membrane by a water- and ion-permeable polyethylene terephthalate (PET mesh. The sensor set-up was tested with egg yolk lecithin liposomes containing gramicidin ion channels and with cell-derived membrane fragments enriched in GABA-gated anion channels. The method allowed monitoring the move of Na(+ and organic cations through gramicidin channels and detecting the Cl(--channel functions of the (α5β2γ2 GABAA receptor in the presence or absence of GABA and the competitive GABA-blocker bicuculline.

  11. Continuous monitoring of dissolved gases with membrane inlet mass spectrometry to fingerprint river biochemical activity

    Science.gov (United States)

    Vautier, Camille; Chatton, Eliot; Abbott, Benjamin; Harjung, Astrid; Labasque, Thierry; Guillou, Aurélie; Pannard, Alexandrine; Piscart, Christophe; Laverman, Anniet; Kolbe, Tamara; Massé, Stéphanie; de Dreuzy, Jean-Raynald; Thomas, Zahra; Aquilina, Luc; Pinay, Gilles

    2017-04-01

    Water quality in rivers results from biogeochemical processes in contributing hydrological compartments (soils, aquifers, hyporheic and riparian zones) and biochemical activity in the river network itself. Consequently, chemical fluxes fluctuate on multiple spatial and temporal scales, leading eventually to complex concentration signals in rivers. We characterized these fluctuations with innovative continuous monitoring of dissolved gases, to quantify transport and reaction processes occurring in different hydrological compartments. We performed stream-scale experiments in two headwater streams in Brittany, France. Factorial injections of inorganic nitrogen (NH4NO3), inorganic phosphate (P2O5) and multiple sources of labile carbon (acetate, tryptophan) were implemented in the two streams. We used a new field application of membrane inlet mass spectrometry to continuously monitor dissolved gases for multiple day-night periods (Chatton et al., 2016). Quantified gases included He, O2, N2, CO2, CH4, N2O, and 15N of dissolved N2 and N2O. We calibrated and assessed the methodology with well-established complementary techniques including gas chromatography and high-frequency water quality sensors. Wet chemistry and radon analysis complemented the study. The analyses provided several methodological and ecological insights and demonstrated that high frequency variations linked to background noise can be efficiently determined and filtered to derive effective fluxes. From a more fundamental point of view, the tested stream segments were fully characterized with extensive sampling of riverbeds and laboratory experiments, allowing scaling of point-level microbial and invertebrate diversity and activity on in-stream processing. This innovative technology allows fully-controlled in-situ experiments providing rich information with a high signal to noise ratio. We present the integrated nutrient demand and uptake and discuss limiting processes and elements at the reach and

  12. Biological activities of peptide concentrates obtained from hydrolysed eggshell membrane byproduct by optimisation with response surface methodology.

    Science.gov (United States)

    Santana, Ana; Melo, Armindo; Tavares, Tânia; Ferreira, Isabel M P L V O

    2016-11-09

    The increase of hen egg consumption demands profitable applications for eggshells, including their membranes, in order to minimize environmental and public health problems that could result from their accumulation. This work presents an innovative application for eggshell membranes to obtain an added-value food ingredient that combines maximized ACE-inhibitory and antioxidant activities. Firstly, the use of acetic acid 5% (v/v); and 3-mercaptopropionic acid 1.25 M enabled 63% recovery of eggshell membrane proteins. Secondly, the extracted proteins were hydrolysed by alcalase from Bacillus licheniformis, viscozyme L and protease from Bacillus amyloliquefaciens. Hydrolysis conditions were optimized using response surface methodology experimental design. The ACE-inhibitory activity (IC 50 ) was 34.5 ± 2.1 μg mL -1 , 63.0 ± 4.2 μg mL -1 and 43.0 ± 8.5 μg mL -1 for each enzyme, respectively, and the antioxidant activity was ca. 4.0 μmol trolox equivalent mg -1 hydrolysed protein . The combination of both bioactive properties is of potential interest to control cardiovascular diseases.

  13. Solubilization, partial purification, and reconstitution of glutamate- and N-methyl-D-aspartate-activated cation channels from brain synaptic membranes

    International Nuclear Information System (INIS)

    Ly, A.M.; Michaelis, E.K.

    1991-01-01

    L-Glutamate-activated cation channel proteins from rat brain synaptic membranes were solubilized, partially purified, and reconstituted into liposomes. Optimal conditions for solubilization and reconstitution included treatment of the membranes with nonionic detergents in the presence of neutral phospholipids plus glycerol. Quench-flow procedures were developed to characterize the rapid kinetics of ion flux induced by receptor agonists. [ 14 C]Methylamine, a cation that permeates through the open channel of both vertebrate and invertebrate glutamate receptors, was used to measure the activity of glutamate receptor-ion channel complexes in reconstituted liposomes. L-Glutamate caused an increase in the rate of [ 14 C]methylamine influx into liposomes reconstituted with either solubilized membrane proteins or partially purified glutamate-binding proteins. Of the major glutamate receptor agonists, only N-methyl-D-aspartate activated cation fluxes in liposomes reconstituted with glutamate-binding proteins. In liposomes reconstituted with glutamate-binding proteins, N-methyl-D-aspartate- or glutamate-induced influx of NA + led to a transient increase in the influx of the lipid-permeable anion probe S 14 CN - . These results indicate the functional reconstitution of N-methyl-D-aspartate-sensitive glutamate receptors and the role of the ∼69-kDa protein in the function of these ion channels

  14. Antifungal Activity of Isoliquiritin and Its Inhibitory Effect against Peronophythora litchi Chen through a Membrane Damage Mechanism

    Directory of Open Access Journals (Sweden)

    Jianjun Luo

    2016-02-01

    Full Text Available This study investigated the antifungal activity and potential antifungal mechanism(s of isoliquiritin against P. litchi Chen, one of the main litchi pathogens. The antifungal activity of isoliquiritin against P. litchi Chen had been proven in a dose-dependent manner through in vitro (mycelial growth and sporangia germination and in vivo (detached leaf tests. Results revealed that isoliquiritin exhibited significant antifungal activity against the tested pathogens, especially, P. litchi Chen, with a minimum inhibitory concentration of 27.33 mg/L. The morphology of P. litchi Chen was apparently changed by isoliquiritin through cytoplasm leakage and distortion of mycelia. The cell membrane permeability of the P. litchi Chen increased with the increasing concentration of isoliquiritin, as evidenced by a rise in relative electric conductivity and a decrease in reducing sugar contents. These results indicated that the antifungal effects of isoliquiritin could be explained by a membrane lesion mechanism causing damage to the cell membrane integrity leading to the death of mycelial cells. Taken together, isoliquiritin may be used as a natural alternative to commercial fungicides or a lead compound to develop new fungicides for the control of litchi downy blight.

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

    Energy Technology Data Exchange (ETDEWEB)

    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 (Helsinki); (Penn)

    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.

  16. Microbial activity catalyzes oxygen transfer in membrane-aerated nitritating biofilm reactors

    DEFF Research Database (Denmark)

    Pellicer i Nàcher, Carles; Domingo Felez, Carlos; Lackner, Susanne

    2013-01-01

    The remarkable oxygen transfer efficiencies attainable in membrane-aerated biofilm reactors (MABRs) are expected to favor their prompt industrial implementation. However, tests in clean water, currently used for the estimation of their oxygen transfer potential, lead to wrong estimates once biofilm...... is present, significantly complicating reactor modelling and control. This study shows for the first time the factors affecting oxygen mass transfer across membranes during clean water tests and reactor operation via undisturbed microelectrode inspection and bulk measurements. The mass transfer resistance...... of the liquid boundary layer developed at the membrane-liquid interface during clean water tests accounted for two thirds of the total mass transfer resistance, suggesting a strong underestimation of the oxygen transfer rates when it is absent (e.g. after biofilm growth). Reactor operation to attain partial...

  17. para-Sulphonato-calix[n]arenes as selective activators for the passage of molecules across the Caco-2 model intestinal membrane.

    Science.gov (United States)

    Roka, Eszter; Vecsernyes, Miklos; Bacskay, Ildiko; Félix, Caroline; Rhimi, Moez; Coleman, Anthony W; Perret, Florent

    2015-06-07

    The passage of Lucifer Yellow across the Caco-2 intestinal model membrane has been studied for the para-sulphonato-calix[n]arenes, the results show that para-sulphonato-calix[4]arene and para-sulphonato-calix[8]arene activate membrane passage when used simultaneously with a transport probe, Lucifer Yellow, whereas para-sulphonato-calix[6]arene has no effect.

  18. Efficiency of membrane technology, activated charcoal, and a micelle-clay complex for removal of the acidic pharmaceutical mefenamic acid.

    Science.gov (United States)

    Khalaf, Samer; Al-Rimawi, Fuad; Khamis, Mustafa; Nir, Shlomo; Bufo, Sabino A; Scrano, Laura; Mecca, Gennaro; Karaman, Rafik

    2013-01-01

    The efficiency of sequential advanced membrane technology wastewater treatment plant towards removal of a widely used non-steroid anti-inflammatory drug (NSAID) mefenamic acid was investigated. The sequential system included activated sludge, ultrafiltration by hollow fibre membranes with 100 kDa cutoff, and spiral wound membranes with 20 kDa cutoff, activated carbon and a reverse osmosis (RO) unit. The performance of the integrated plant showed complete removal of mefenamic acid from spiked wastewater samples. The activated carbon column was the most effective component in removing mefenamic acid with a removal efficiency of 97.2%. Stability study of mefenamic acid in pure water and Al-Quds activated sludge revealed that the anti-inflammatory drug was resistant to degradation in both environments. Batch adsorption of mefenamic acid by activated charcoal and a composite micelle (otadecyltrimethylammonium (ODTMA)-clay (montmorillonite) was determined at 25.0°C. Langmuir isotherm was found to fit the data with Qmax of 90.9 mg g(-1) and 100.0 mg g(-1) for activated carbon and micelle-clay complex, respectively. Filtration experiment by micelle-clay columns mixed with sand in the mg L(-1) range revealed complete removal of the drug with much larger capacity than activated carbon column. The combined results demonstrated that an integration of a micelle-clay column in the plant system has a good potential to improve the removal efficiency of the plant towards NSAID drugs such as mefenamic acid.

  19. Depression of membrane-bound Na sup + -K sup + -ATPase activity induced by free radicals and by ischemia of kidney

    Energy Technology Data Exchange (ETDEWEB)

    Kako, K.; Kato, M.; Matsuoka, T.; Mustapha, A. (Univ. of Ottawa, Ontario (Canada))

    1988-02-01

    A partially purified, membrane-bound Na{sup +}-K{sup +}-ATPase fraction, prepared from the outer medulla of porcine kidney, was incubated in the presence of 0.1-100 mM H{sub 2}O{sub 2} for either 15 or 30 min at 37{degree}C. The activity of ouabain-sensitive Na{sup +}-K{sup +}-ATPase was reduced proportionally to the concentration of H{sub 2}O{sub 2} and the duration of incubation. There were decreases in SH contents and turnover rates of the Na{sup +}-K{sup +}-ATPase preparation, while malondialdehyde (MDA) and conjugated dienes were generated from the membrane lipids in the course of the incubation. The concentrations of ethanolamine (E) plasmalogen and of arachidonic acid in the E glycerophospholipid molecules were reduced by the free radical reaction. Similarly, a reduction in Na{sup +}K{sup +}-ATPase activity and the formation of MDA and conjugated dienes, together with a decrease in E glycerophospholipids, were observed when the membrane fraction was exposed to ultraviolet irradiation (254 nm) for 30 min at 4{degree}C. Microsomal fractions, prepared from the outer medulla of canine kidney after 1 h of unilateral ischemia and 1 h of reperfusion, showed a decreased Na{sup +}-K{sup +}-ATPase activity, a reduced amount of SH groups, and an increased MDA. These changes were normalized by the infusion of N-mercaptopropionylglycine. These results support the view (1) that free radical generation affects the enzyme protein as well as membrane lipids, and (2) that free radicals may be formed in the ischemic reperfused kidney.

  20. Persistent alterations in active and passive electrical membrane properties of regenerated nerve fibers of man and mice.

    Science.gov (United States)

    Moldovan, Mihai; Alvarez, Susana; Rosberg, Mette R; Krarup, Christian

    2016-02-01

    Excitability of regenerated fibers remains impaired due to changes in both passive cable properties and alterations in the voltage-dependent membrane function. These abnormalities were studied by mathematical modeling in human regenerated nerves and experimental studies in mice. In three adult male patients with surgically repaired complete injuries of peripheral nerves of the arm 22 months-26 years prior to investigation, deviation of excitability measures was explained by a hyperpolarizing shift in the resting membrane potential and an increase in the passive 'Barrett and Barrett' conductance (GBB) bridging the nodal and internodal compartments. These changes were associated with an increase in the 'fast' K(+) conductance and the inward rectifier conductance (GH). Similar changes were found in regenerated mouse tibial motor axons at 1 month after a sciatic crush lesion. During the first 5 months of regeneration, GH showed partial recovery, which paralleled that in GBB. The internodal length remained one-third of normal. Excitability abnormalities could be reversed by the energy-dependent Na(+)/K(+) pump blocker ouabain resulting in membrane depolarization. Stressing the Na(+) pumping system during a strenuous activity protocol triggered partial Wallerian degeneration in regenerated nerves but not in control nerves from age-matched mice. The current data suggest that the nodal voltage-gated ion channel machinery is restored in regenerated axons, although the electrical separation from the internodal compartment remains compromised. Due to the persistent increase in number of nodes, the increased activity-dependent Na(+) influx could lead to hyperactivity of the Na(+)/K(+) pump resulting in membrane hyperpolarization and neurotoxic energy insufficiency during strenuous activity. © 2015 Federation of European Neuroscience Societies and John Wiley & Sons Ltd.

  1. Release of LHRH-activity from human fetal membranes upon exposure to PGE/sub 2/, oxytocin and isoproterenol

    Energy Technology Data Exchange (ETDEWEB)

    Poisner, A.M.; Poisner, R.; Becca, C.R.; Conn, P.M.

    1986-03-01

    The authors have previously reported that superfused chorion laeve (fetal membranes) release LHRH-like immunoreactivity upon exposure to angiotensin II. They have now studied the effects of other agonists on the release of LHRH-activity and something of its chemical nature. Fetal membranes were obtained from placentas delivered by cesarean section, the amnion stripped from the chorion, and the chorion superfused in an Amicon thin-channel device with the maternal surface facing up. The whole device was submerged in a 37 C water bath and perfused with a modified Locke's solution at 0.4 - 1.0 ml/min. LHRH-activity was measured by radioimmunoassay using three different antisera against LHRH. The release of LHRH-activity was stimulated by 6-10 min exposure to PGE/sub 2/, oxytocin, and isoproterenol. Extracts of chorion were studied using gel filtration on Sephacryl S-200 and ultrafiltration with Amicon PM-10 filters. The bulk of the LHRH-activity appeared as a higher molecular weight form (about 70,000 daltons). Since oxytocin has been reported to release PGE/sub 2/ from chorion, it may release LHRH-activity by virtue of liberating endogenous PGE/sub 2/. The chemical nature of the LHRH-activity is presently under investigation.

  2. Development of a Simulation Model for P Systems With Active Membranes

    Science.gov (United States)

    2006-07-01

    in the membrane computing area. The tool is user friendly, allowing the user to follow the evolution of a P system in a visual way. The simulator can...Mitrana, G. Paun, On the power of P systems with valuations, Computacion Sistemas 5 (2001) pp. 120-127.

  3. Functional activity of Gi alpha protein in detergent resistant membrane domains from rat brain cortex

    Czech Academy of Sciences Publication Activity Database

    Stöhr, Jiří; Rudajev, Vladimír; Bouřová, Lenka; Lisý, Václav; Novotný, Jiří; Svoboda, Petr

    2007-01-01

    Roč. 101, Suppl.1 (2007), s. 52-52 ISSN 0022-3042. [European Society for Neurochemistry Meeting /17./. 19.05.2007-22.05.2007, Salamanca] Institutional research plan: CEZ:AV0Z50110509 Keywords : cpo1 * GABAB receptor * Gi protein * membrane domains Subject RIV: ED - Physiology

  4. In vitro and in vivo activation of mitochondrial membrane permeability transition pore using triiodothyronine

    Czech Academy of Sciences Publication Activity Database

    Endlicher, R.; Drahota, Zdeněk; Červinková, Z.

    2016-01-01

    Roč. 65, č. 2 (2016), s. 321-331 ISSN 0862-8408 R&D Projects: GA ČR(CZ) GB14-36804G Institutional support: RVO:67985823 Keywords : rat liver mitochondria * membrane permeability transition pore * thyroid hormones Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 1.461, year: 2016

  5. Dipolar Relaxation Dynamics at the Active Site of an ATPase Regulated by Membrane Lateral Pressure

    Czech Academy of Sciences Publication Activity Database

    Fischermeier, E.; Pospíšil, Petr; Sayed, A.; Hof, Martin; Solioz, M.; Fahmy, K.

    2017-01-01

    Roč. 56, č. 5 (2017), s. 1269-1272 ISSN 1433-7851 R&D Projects: GA ČR(CZ) GBP208/12/G016 Institutional support: RVO:61388955 Keywords : fluorescence * ion pump * membrane proteins * nanodiscs * time-resolved emission Subject RIV: CF - Physical ; Theoretical Chemistry OBOR OECD: Physical chemistry Impact factor: 11.994, year: 2016

  6. Membrane plasmalogen composition and cellular cholesterol regulation: a structure activity study

    Directory of Open Access Journals (Sweden)

    Su-Myat Khine K

    2010-06-01

    Full Text Available Abstract Background Disrupted cholesterol regulation leading to increased circulating and membrane cholesterol levels is implicated in many age-related chronic diseases such as cardiovascular disease (CVD, Alzheimer's disease (AD, and cancer. In vitro and ex vivo cellular plasmalogen deficiency models have been shown to exhibit impaired intra- and extra-cellular processing of cholesterol. Furthermore, depleted brain plasmalogens have been implicated in AD and serum plasmalogen deficiencies have been linked to AD, CVD, and cancer. Results Using plasmalogen deficient (NRel-4 and plasmalogen sufficient (HEK293 cells we investigated the effect of species-dependent plasmalogen restoration/augmentation on membrane cholesterol processing. The results of these studies indicate that the esterification of cholesterol is dependent upon the amount of polyunsaturated fatty acid (PUFA-containing ethanolamine plasmalogen (PlsEtn present in the membrane. We further elucidate that the concentration-dependent increase in esterified cholesterol observed with PUFA-PlsEtn was due to a concentration-dependent increase in sterol-O-acyltransferase-1 (SOAT1 levels, an observation not reproduced by 3-hydroxy-3-methyl-glutaryl-CoA (HMG-CoA reductase inhibition. Conclusion The present study describes a novel mechanism of cholesterol regulation that is consistent with clinical and epidemiological studies of cholesterol, aging and disease. Specifically, the present study describes how selective membrane PUFA-PlsEtn enhancement can be achieved using 1-alkyl-2-PUFA glycerols and through this action reduce levels of total and free cholesterol in cells.

  7. Cathepsin activities and membrane integrity of zebrafish (Danio rerio) oocytes after freezing to -196 degrees C using controlled slow cooling.

    Science.gov (United States)

    Zhang, T; Rawson, D M; Tosti, L; Carnevali, O

    2008-04-01

    This study investigated enzymatic activity of cathepsins and the membrane integrity of zebrafish (Danio rerio) oocytes after freezing to -196 degrees C using controlled slow cooling. Stage III oocytes (>0.5mm), obtained through dissection of anaesthetised female fish and desegregation of ovarian cumulus, were exposed to 2M methanol or 2M DMSO (both prepared in Hank's medium) for 30min at 22 degrees C before being loaded into 0.5ml plastic straws and placed into a programmable cooler. After controlled slow freezing, samples were plunged into liquid nitrogen (LN) and held for at least 10min, and thawed by immersing straws into a 27 degrees C water bath for 10s. Thawed oocytes were washed twice in Hank's medium. Cathepsin activity and membrane integrity of oocytes were assessed both after cryoprotectant treatment at 22 degrees C and after freezing in LN. Cathepsin B and L colorimetric analyses were performed using substrates Z-Arg-ArgNNap and Z-Phe-Arg-4MbetaNA-HCl, respectively, and 2-naphthylamine and 4-methoxy-2-naphthylamine were used as standards. Cathepsin D activity was performed by analysing the level of hydrolytic action on haemoglobin. Oocytes membrane integrity was assessed using 0.2% Trypan blue staining for 5min. Analysis of cathepsin activities showed that whilst the activity of cathepsin B and D was not affected by 2M DMSO treatment, their activity was lowered when treated with 2M methanol. Following freezing to -196 degrees C, the activity of all cathepsins (B, D and L) was significantly decreased in both 2M DMSO and 2M methanol. Trypan blue staining showed that 63.0+/-11.3% and 72.7+/-5.2% oocytes membrane stayed intact after DMSO and methanol treatment for 30min at 22 degrees C, respectively, whilst 14.9+/-2.6% and 1.4+/-0.8% stayed intact after freezing in DMSO and methanol to -196 degrees C. The results indicate that cryoprotectant treatment and freezing modified the activities of lysosomal enzymes involved in oocyte maturation and yolk

  8. ATPase activity of erythrocyte membranes and their permeability for the K-ions as influenced by irradiation and serotonin

    International Nuclear Information System (INIS)

    Zhegnevskaya, V.V.; Vinogradova, M.F.; Polevoj, V.V.

    1982-01-01

    Na, K-ATPase activity of membranes of erytrocytes after 1 hour of X-ray irradiation of citrate blood of rats (25.8 Kl/kg)-increased, and after irradiation of isolated erytrocytes, placed in the isotonic solution of NaCl did not change. The exflux of K-ions out of irradiated erytrocytes increased equally in both cases. Serotonin (2x10 -4 M), added to the probes 10 minutes before irradiation, decreased the exflux of K + by irradiated erytrocytes, but Na, K-ATPase activity under the influence of amine was without changes

  9. Isolation and Molecular Characterization of Biofouling Bacteria and Profiling of Quorum Sensing Signal Molecules from Membrane Bioreactor Activated Sludge

    Directory of Open Access Journals (Sweden)

    Harshad Lade

    2014-02-01

    Full Text Available The formation of biofilm in a membrane bioreactor depends on the production of various signaling molecules like N-acyl homoserine lactones (AHLs. In the present study, a total of 200 bacterial strains were isolated from membrane bioreactor activated sludge and screened for AHLs production using two biosensor systems, Chromobacterium violaceum CV026 and Agrobacterium tumefaciens A136. A correlation between AHLs production and biofilm formation has been made among screened AHLs producing strains. The 16S rRNA gene sequence analysis revealed the dominance of Aeromonas and Enterobacter sp. in AHLs production; however few a species of Serratia, Leclercia, Pseudomonas, Klebsiella, Raoultella and Citrobacter were also identified. The chromatographic characterization of sludge extract showed the presence of a broad range of quorum sensing signal molecules. Further identification of sludge AHLs by thin layer chromatography bioassay and high performance liquid chromatography confirms the presence of C4-HSL, C6-HSL, C8-HSL, 3-oxo-C8-HSL, C10-HSL, C12-HSL, 3-oxo-C12-HSL and C14-HSL. The occurrence of AHLs in sludge extract and dominance of Aeromonas and Enterobacter sp. in activated sludge suggests the key role of these bacterial strains in AHLs production and thereby membrane fouling.

  10. Isolation and molecular characterization of biofouling bacteria and profiling of quorum sensing signal molecules from membrane bioreactor activated sludge.

    Science.gov (United States)

    Lade, Harshad; Paul, Diby; Kweon, Ji Hyang

    2014-02-04

    The formation of biofilm in a membrane bioreactor depends on the production of various signaling molecules like N-acyl homoserine lactones (AHLs). In the present study, a total of 200 bacterial strains were isolated from membrane bioreactor activated sludge and screened for AHLs production using two biosensor systems, Chromobacterium violaceum CV026 and Agrobacterium tumefaciens A136. A correlation between AHLs production and biofilm formation has been made among screened AHLs producing strains. The 16S rRNA gene sequence analysis revealed the dominance of Aeromonas and Enterobacter sp. in AHLs production; however few a species of Serratia, Leclercia, Pseudomonas, Klebsiella, Raoultella and Citrobacter were also identified. The chromatographic characterization of sludge extract showed the presence of a broad range of quorum sensing signal molecules. Further identification of sludge AHLs by thin layer chromatography bioassay and high performance liquid chromatography confirms the presence of C4-HSL, C6-HSL, C8-HSL, 3-oxo-C8-HSL, C10-HSL, C12-HSL, 3-oxo-C12-HSL and C14-HSL. The occurrence of AHLs in sludge extract and dominance of Aeromonas and Enterobacter sp. in activated sludge suggests the key role of these bacterial strains in AHLs production and thereby membrane fouling.

  11. Mg(2+,ATP-dependent plasma membrane calcium pump of smooth muscle cells. ІІ. Regulation of activity

    Directory of Open Access Journals (Sweden)

    T. О. Veklich

    2015-04-01

    Full Text Available Plasma membrane Ca2+-pump is one of key proteins, which takes part in Ca2+ exchange in smooth muscle cells. It has a lot of diverse functions from control of basal cytoplasmal Ca2+ concentration to regulation of proteins involved in Ca2+-dependent signal pathway. Ca2+ pump function is often depen­dent on the isoform or even form of alternative splicing. Allowing for a variety of Ca2+-pump functions and properties, which were reviewed in detail in the first part of our review article cycle (Ukr. Biochem. J., 2015; 87(1, the precise control of the mentioned pump activity is very important for cell functioning­. The other part of this article is dedicated to different regulation factors of smooth muscle plasma membrane Ca2+-pump activity: endogenous and exo­genous, biotic and abiotic factors. Special attention is given to literature data and own results about design and the search of selective plasma membrane Ca2+-pump inhibitor which would allow examining its functioning in smooth muscle cells more meticulously.

  12. Membrane-active macromolecules kill antibiotic-tolerant bacteria and potentiate antibiotics towards Gram-negative bacteria.

    Directory of Open Access Journals (Sweden)

    Divakara S S M Uppu

    Full Text Available Chronic bacterial biofilms place a massive burden on healthcare due to the presence of antibiotic-tolerant dormant bacteria. Some of the conventional antibiotics such as erythromycin, vancomycin, linezolid, rifampicin etc. are inherently ineffective against Gram-negative bacteria, particularly in their biofilms. Here, we report membrane-active macromolecules that kill slow dividing stationary-phase and antibiotic tolerant cells of Gram-negative bacteria. More importantly, these molecules potentiate antibiotics (erythromycin and rifampicin to biofilms of Gram-negative bacteria. These molecules eliminate planktonic bacteria that are liberated after dispersion of biofilms (dispersed cells. The membrane-active mechanism of these molecules forms the key for potentiating the established antibiotics. Further, we demonstrate that the combination of macromolecules and antibiotics significantly reduces bacterial burden in mouse burn and surgical wound infection models caused by Acinetobacter baumannii and Carbapenemase producing Klebsiella pneumoniae (KPC clinical isolate respectively. Colistin, a well-known antibiotic targeting the lipopolysaccharide (LPS of Gram-negative bacteria fails to kill antibiotic tolerant cells and dispersed cells (from biofilms and bacteria develop resistance to it. On the contrary, these macromolecules prevent or delay the development of bacterial resistance to known antibiotics. Our findings emphasize the potential of targeting the bacterial membrane in antibiotic potentiation for disruption of biofilms and suggest a promising strategy towards developing therapies for topical treatment of Gram-negative infections.

  13. Membrane-active macromolecules kill antibiotic-tolerant bacteria and potentiate antibiotics towards Gram-negative bacteria.

    Science.gov (United States)

    Uppu, Divakara S S M; Konai, Mohini M; Sarkar, Paramita; Samaddar, Sandip; Fensterseifer, Isabel C M; Farias-Junior, Celio; Krishnamoorthy, Paramanandam; Shome, Bibek R; Franco, Octávio L; Haldar, Jayanta

    2017-01-01

    Chronic bacterial biofilms place a massive burden on healthcare due to the presence of antibiotic-tolerant dormant bacteria. Some of the conventional antibiotics such as erythromycin, vancomycin, linezolid, rifampicin etc. are inherently ineffective against Gram-negative bacteria, particularly in their biofilms. Here, we report membrane-active macromolecules that kill slow dividing stationary-phase and antibiotic tolerant cells of Gram-negative bacteria. More importantly, these molecules potentiate antibiotics (erythromycin and rifampicin) to biofilms of Gram-negative bacteria. These molecules eliminate planktonic bacteria that are liberated after dispersion of biofilms (dispersed cells). The membrane-active mechanism of these molecules forms the key for potentiating the established antibiotics. Further, we demonstrate that the combination of macromolecules and antibiotics significantly reduces bacterial burden in mouse burn and surgical wound infection models caused by Acinetobacter baumannii and Carbapenemase producing Klebsiella pneumoniae (KPC) clinical isolate respectively. Colistin, a well-known antibiotic targeting the lipopolysaccharide (LPS) of Gram-negative bacteria fails to kill antibiotic tolerant cells and dispersed cells (from biofilms) and bacteria develop resistance to it. On the contrary, these macromolecules prevent or delay the development of bacterial resistance to known antibiotics. Our findings emphasize the potential of targeting the bacterial membrane in antibiotic potentiation for disruption of biofilms and suggest a promising strategy towards developing therapies for topical treatment of Gram-negative infections.

  14. ECUT: Energy Conversion and Utilization Technologies program biocatalysis research activity. Potential membrane applications to biocatalyzed processes: Assessment of concentration polarization and membrane fouling

    Science.gov (United States)

    Ingham, J. D.

    1983-01-01

    Separation and purification of the products of biocatalyzed fermentation processes, such as ethanol or butanol, consumes most of the process energy required. Since membrane systems require substantially less energy for separation than most alternatives (e.g., distillation) they have been suggested for separation or concentration of fermentation products. This report is a review of the effects of concentration polarization and membrane fouling for the principal membrane processes: microfiltration, ultrafiltration, reverse osmosis, and electrodialysis including a discussion of potential problems relevant to separation of fermentation products. It was concluded that advanced membrane systems may result in significantly decreased energy consumption. However, because of the need to separate large amounts of water from much smaller amounts of product that may be more volatile than wate, it is not clear that membrane separations will necessarily be more efficient than alternative processes.

  15. Active Fault Geometry and Crustal Deformation Along the San Andreas Fault System Through San Gorgonio Pass, California: The View in 3D From Seismicity

    Science.gov (United States)

    Nicholson, C.; Hauksson, E.; Plesch, A.

    2012-12-01

    Understanding the 3D geometry and deformation style of the San Andreas fault (SAF) is critical to accurate dynamic rupture and ground motion prediction models. We use 3D alignments of hypocenter and focal mechanism nodal planes within a relocated earthquake catalog (1981-2011) [Hauksson et al., 2012] to develop improved 3D fault models for active strands of the SAF and adjacent secondary structures. Through San Gorgonio Pass (SGP), earthquakes define a mechanically layered crust with predominantly high-angle strike-slip faults in the upper ~10 km, while at greater depth, intersecting sets of strike-slip, oblique slip and low-angle thrust faults define a wedge-shaped volume deformation of the lower crust. In some places, this interface between upper and lower crustal deformation may be an active detachment fault, and may have controlled the down-dip extent of recent fault rupture. Alignments of hypocenters and nodal planes define multiple principal slip surfaces through SGP, including a through-going steeply-dipping predominantly strike-slip Banning fault strand at depth that upward truncates a more moderately dipping (40°-50°) blind, oblique North Palm Springs fault. The North Palm Springs fault may be the active down-dip extension of the San Gorgonio Pass thrust offset at depth by the principal, through-going Banning strand. In the northern Coachella Valley, seismicity indicates that the Garnet Hill and Banning fault strands are most likely sub-parallel and steeply dipping (~70°NE) to depths of 8-10 km, where they intersect and merge with a stack of moderately dipping to low-angle oblique thrust faults. Gravity and water well data confirm that these faults are sub-parallel and near vertical in the upper 2-3 km. Although the dense wedge of deep seismicity below SGP and largely south of the SAF contains multiple secondary fault sets of different orientations, the predominant fault set appears to be a series of en echelon NW-striking oblique strike-slip faults

  16. Important factors for cell-membrane permeabilization by gold nanoparticles activated by nanosecond-laser irradiation

    Directory of Open Access Journals (Sweden)

    Yao CP

    2017-08-01

    Full Text Available Cuiping Yao,1,2,* Florian Rudnitzki,2,* Gereon Hüttmann,2,3 Zhenxi Zhang,1 Ramtin Rahmanzadeh2 1Key Laboratory of Biomedical Information Engineering of Education Ministry, Institute of Biomedical Analytical Technology and Instrumentation, School of Life Science and Technology, Xi’an Jiaotong University, Xi’an, China; 2Institute of Biomedical Optics, University of Lübeck, Lübeck, 3Airway Research Center North (ARCN, Member of the German Center for Lung Research (DZL, Kiel, Germany *These authors contributed equally to this work Purpose: Pulsed-laser irradiation of light-absorbing gold nanoparticles (AuNPs attached to cells transiently increases cell membrane permeability for targeted molecule delivery. Here, we targeted EGFR on the ovarian carcinoma cell line OVCAR-3 with AuNPs. In order to optimize membrane permeability and to demonstrate molecule delivery into adherent OVCAR-3 cells, we systematically investigated different experimental conditions. Materials and methods: AuNPs (30 nm were functionalized by conjugation of the antibody cetuximab against EGFR. Selective binding of the particles was demonstrated by silver staining, multiphoton imaging, and fluorescence-lifetime imaging. After laser irradiation, membrane permeability of OVCAR-3 cells was studied under different conditions of AuNP concentration, cell-incubation medium, and cell–AuNP incubation time. Membrane permeability and cell viability were evaluated by flow cytometry, measuring propidium iodide and fluorescein isothiocyanate–dextran uptake. Results: Adherently growing OVCAR-3 cells can be effectively targeted with EGFR-AuNP. Laser irradiation led to successful permeabilization, and 150 kDa dextran was successfully delivered into cells with about 70% efficiency. Conclusion: Antibody-targeted and laser-irradiated AuNPs can be used to deliver molecules into adherent cells. Efficacy depends not only on laser parameters but also on AuNP:cell ratio, cell-incubation medium

  17. Earliest Results in the Use of Activated Composite Membranes for the Transport of Silver Ions from Aqueous Solutions

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    Yucundo Mendoza-Tolentino

    2014-01-01

    Full Text Available This paper presents the results concerning the first use of activated composite membranes (ACMs for the facilitated transport of silver ions containing di-(2-ethylhexyl-dithiophosphoric acid (DTPA as the carrier. DTPA was immobilized by interfacial polymerization in a dense layer that was deposited in a porous layer, which was prepared on a nonwoven fabric support by phase inversion. The influence of fundamental parameters affecting the transport of silver ion as the carrier concentration in the membrane phase and stripping agent variation of the stripping solution have been studied. In the optimal conditions, the amount of silver transported across the ACMs was greater than 50%, whereas if the content of the carrier is modified, more than the 90% of the initial silver is removed from the feed phase.

  18. Persistent alterations in active and passive electrical membrane properties of regenerated nerve fibers of man and mice

    DEFF Research Database (Denmark)

    Moldovan, Mihai; Alvarez Herrero, Susana; Rosberg, Mette R.

    2016-01-01

    patients with surgically repaired complete injuries of peripheral nerves of the arm 22 months-26 years prior to investigation, deviation of excitability measures was explained by a hyperpolarizing shift in the resting membrane potential and an increase in the passive 'Barrett and Barrett' conductance (GBB......Excitability of regenerated fibers remains impaired due to changes in both passive cable properties and alterations in the voltage-dependent membrane function. These abnormalities were studied by mathematical modeling in human regenerated nerves and experimental studies in mice. In three adult male...... activity protocol triggered partial Wallerian degeneration in regenerated nerves but not in control nerves from age-matched mice. The current data suggest that the nodal voltage-gated ion channel machinery is restored in regenerated axons, although the electrical separation from the internodal compartment...

  19. The ferric-reducing activity of duodenal brush-border membrane vesicles is associated with a b-type haem.

    Science.gov (United States)

    Pountney, D J; Raja, K B; Simpson, R J; Wrigglesworth, J M

    1999-03-01

    Rabbit brush-border membrane vesicles possess ferricyanide reducing activity. This activity is preferentially dependent on NADH as reductant, and can be stimulated by the addition of FMN. The latency of activity observed following vesicle solubilisation suggests that the responsible component is transmembranous, and partially sequestered on the inner-face of the vesicles prior to full solubilisation. Subsequent increases in detergent concentration (> 0.3% w/v lauryl maltoside) were found to be inhibitory. Ferricyanide reducing activity was effectively inhibited by the sulphydryl modifying reagents N-ethyl malemide and p-chloromercuribenzoate, but not by the flavin analogue diphenylene iodonium. The ferric-reducing activity co-purified with a b-type haem when applied to Sephacryl S-200 columns. The putative cytochrome was found to be immunologically distinct from neutrophil cytochrome b558.

  20. Osmolality, temperature, and membrane lipid composition modulate the activity of betaine transporter BetP in Corynebacterium glutamicum

    DEFF Research Database (Denmark)

    Ozcan, Nuran; Ejsing, Christer S.; Shevchenko, Andrej

    2007-01-01

    , BetP, is instantly activated in response to an increasing cytoplasmic K(+) concentration. Importantly, it is also activated by chill stress independent of osmotic stress. We show that the activation of BetP by both osmotic stress and chill stress is altered in C. glutamicum cells grown at and adapted...... to low temperatures. BetP from cold-adapted cells is less sensitive to osmotic stress. In order to become susceptible for chill activation, cold-adapted cells in addition needed a certain amount of osmotic stimulation, indicating that there is cross talk of these two types of stimuli at the level of Bet......P activity. We further correlated the change in BetP regulation properties in cells grown at different temperatures to changes in the lipid composition of the plasma membrane. For this purpose, the glycerophospholipidome of C. glutamicum grown at different temperatures was analyzed by mass spectrometry using...

  1. Alterations in hemostatic parameters during hemodialysis with dialyzers of different membrane composition and flow design. Platelet activation and factor VIII-related von Willebrand factor during hemodialysis.

    Science.gov (United States)

    Schmitt, G W; Moake, J L; Rudy, C K; Vicks, S L; Hamburger, R J

    1987-09-01

    The effect of dialyzer membrane and design on hemostatic parameters during hemodialysis were evaluated in a prospective controlled study. This study demonstrated that hemodialysis is associated with significant platelet activation and loss, which are influenced by both dialyzer configuration and membrane composition. In addition, use of the cuprophan membrane is associated with greater perturbations of the vascular endothelium, as reflected in changes in factor VIII-related von Willebrand factor and 6-keto-prostaglandin F1 alpha concentrations not seen with the polyacrylonitrile membrane. Of the dialyzers studied, the polyacrylonitrile membrane in a hollow-fiber configuration appears to minimize platelet loss and activation, and to minimize increases in factor VIII-related von Willebrand factor and 6-keto-prostaglandin F1 alpha.

  2. Vertical-axis rotations and deformation along the active strike-slip El Tigre Fault (Precordillera of San Juan, Argentina) assessed through palaeomagnetism and anisotropy of magnetic susceptibility

    Science.gov (United States)

    Fazzito, Sabrina Y.; Rapalini, Augusto E.; Cortés, José M.; Terrizzano, Carla M.

    2017-03-01

    Palaeomagnetic data from poorly consolidated to non-consolidated late Cenozoic sediments along the central segment of the active El Tigre Fault (Central-Western Precordillera of the San Juan Province, Argentina) demonstrate broad cumulative deformation up to 450 m from the fault trace and reveal clockwise and anticlockwise vertical-axis rotations of variable magnitude. This deformation has affected in different amounts Miocene to late Pleistocene samples and indicates a complex kinematic pattern. Several inherited linear structures in the shear zone that are oblique to the El Tigre Fault may have acted as block boundary faults. Displacement along these faults may have resulted in a complex pattern of rotations. The maximum magnitude of rotation is a function of the age of the sediments sampled, with largest values corresponding to middle Miocene-lower Pliocene deposits and minimum values obtained from late Pleistocene deposits. The kinematic study is complemented by low-field anisotropy of magnetic susceptibility data to show that the local strain regime suggests a N-S stretching direction, subparallel to the strike of the main fault.

  3. Intracellular and membrane-damaging activities of methyl gallate isolated from Terminalia chebula against multidrug-resistant Shigella spp.

    Science.gov (United States)

    Acharyya, Saurabh; Sarkar, Prodipta; Saha, Dhira R; Patra, Amarendra; Ramamurthy, T; Bag, Prasanta K

    2015-08-01

    Shigella spp. (Shigella dysenteriae, Shigella flexneri, Shigella boydii and Shigella sonnei) cause bacillary dysentery (shigellosis), which is characterized by bloody mucous diarrhoea. Although a variety of antibiotics have been effective for treatment of shigellosis, options are becoming limited due to globally emerging drug resistance. In the present study, in vitro antibacterial activity of methyl gallate (MG) isolated from Terminalia chebula was determined by performing MIC, minimal bactericidal concentration (MBC) and time-kill kinetic studies. Bacterial membrane-damaging activity of MG was determined by membrane perturbation and transmission electron microscopy (TEM). Cellular drug accumulation, cell infection and assessment of intracellular activities of MG and reference antibiotics were performed using HeLa cell cultures. The bactericidal activity of MG against multidrug-resistant (MDR) Shigella spp. in comparison with other commonly used drugs including fluoroquinolone was demonstrated here. TEM findings in the present study revealed that MG caused the total disintegration of inner and outer membranes, and leakage of the cytoplasmic contents of S. dysenteriae. The level of accumulation of MG and tetracycline in HeLa cells incubated for 24  h was relatively higher than that of ciprofloxacin and nalidixic acid (ratio of intracellular concentration/extracellular concentration of antibiotic for MG and tetracycline>ciprofloxacin and nalidixic acid). The viable number of intracellular S. dysenteriae was decreased in a time-dependent manner in the presence of MG (4 × MBC) and reduced to zero within 20  h. The significant intracellular activities of MG suggested that it could potentially be used as an effective antibacterial agent for the treatment of severe infections caused by MDR Shigella spp.

  4. Activity of hydrolytic enzymes of Candida albicans strains isolated from patients with periodontal and membrane mucosae of oral cavity diseases.

    Science.gov (United States)

    Kurnatowska, A J

    1998-01-01

    Fungi are elements of the ontocenosis of the oral cavity and causal factors of inflammatory lesions in its mucous membrane. The objective of the study was to find differences in the activity of hydrolytic enzymes of Candida albicans isolated from patients with diseases of the periodontium and mucous membrane of the oral cavity. Of 235 patients examined, 31 were diagnosed with gingivitis, 38 with glossitis, 28 with leucoplakia, 37 with adult periodontitis, 25 with juvenile periodontitis, 36 stomatitis prothetica and 40 with stomatitis atrophica. In 196 patients (83.4 +/- 2.4%), fungi belonging to Candida species were detected. In the evaluation of Candida albicans strains (146) properties, bioMerieux API ZYM tests containing substrates for the detection of 19 hydrolases were used. All the investigated strains were characterized by the activity of 14 enzymes, i.e. phosphatase alcaline, esterase (C4), esterase lipase (C8), leucine and valine arylamidase, phosphatase acid, naphthol-AS-BI-phosphohydrolase, alpha galactosidase, beta galactosidase, alpha glucosidase, beta glucosidase, N-acetyl-beta-glucosaminidase, alpha mannosidase and alpha fucosidase. Strains isolated from the oral cavity of patients with disease of periodontium and mucous membrane are characterised by the highest phosphatase acid activity. The greatest enzymatic activity is characteristic of Candida albicans isolated from patients with stomatitis atrophica or stomatitis prothetica, and the lowest in strains from gingivitis or juvenile periodontitis cases. Differences in the activity of hydrolases are statistically significant (p phosphatase acid, naphthol-AS-BI-phosphohydrolase, beta glucosidase, N-acetyl-beta-glucosaminidase, of fungi isolated from patients with particular clinical diagnoses.

  5. A Novel Hierarchical Structured Poly(lactic acid/Titania Fibrous Membrane with Excellent Antibacterial Activity and Air Filtration Performance

    Directory of Open Access Journals (Sweden)

    Zhe Wang

    2016-01-01

    Full Text Available Hybrid poly(lactic acid/titania (PLA/TiO2 fibrous membranes exhibiting excellent air filtration performance and good antibacterial activity were prepared via the electrospinning technique. By varying the composition of the precursor solutions and the relative humidity, the morphologies of PLA/TiO2 fibers, including the nanopores and nanometer-scale protrusions on the surface of the fibers, could be regulated. The distribution of nanopores and TiO2 nanoparticles on the surface of PLA/TiO2 fibers was investigated. Nitrogen adsorption-desorption analysis revealed that nanopores and nanometer-scale protrusions play an important role in improving the specific surface area and nanopore volume of the relevant PLA/TiO2 fibrous membrane. Filtration performance tests conducted by measuring the penetration of sodium chloride aerosol particles with a 260 nm mass median diameter indicated that fibers with a high surface roughness, large specific surface area, and large nanopore volume greatly improved the particle capture efficiency and facilitated the penetration of airflow. Furthermore, the introduction of TiO2 nanoparticles endows the relevant fibrous membrane with antibacterial properties. The as-prepared PLA/TiO2 fibrous membrane loaded with 1.75 wt% TiO2 nanoparticles formed at a relative humidity of 45% exhibited a high filtration efficiency (99.996% and a relatively low pressure drop (128.7 Pa, as well as a high antibacterial activity of 99.5%.

  6. Regulation of Smoothened Phosphorylation and High-Level Hedgehog Signaling Activity by a Plasma Membrane Associated Kinase.

    Directory of Open Access Journals (Sweden)

    Shuangxi Li

    2016-06-01

    Full Text Available Hedgehog (Hh signaling controls embryonic development and adult tissue homeostasis through the G protein coupled receptor (GPCR-family protein Smoothened (Smo. Upon stimulation, Smo accumulates on the cell surface in Drosophila or primary cilia in vertebrates, which is thought to be essential for its activation and function, but the underlying mechanisms remain poorly understood. Here we show that Hh stimulates the binding of Smo to a plasma membrane-associated kinase Gilgamesh (Gish/CK1γ and that Gish fine-tunes Hh pathway activity by phosphorylating a Ser/Thr cluster (CL-II in the juxtamembrane region of Smo carboxyl-terminal intracellular tail (C-tail. We find that CL-II phosphorylation is promoted by protein kinase A (PKA-mediated phosphorylation of Smo C-tail and depends on cell surface localization of both Gish and Smo. Consistent with CL-II being critical for high-threshold Hh target gene expression, its phosphorylation appears to require higher levels of Hh or longer exposure to the same level of Hh than PKA-site phosphorylation on Smo. Furthermore, we find that vertebrate CK1γ is localized at the primary cilium to promote Smo phosphorylation and Sonic hedgehog (Shh pathway activation. Our study reveals a conserved mechanism whereby Hh induces a change in Smo subcellular localization to promote its association with and activation by a plasma membrane localized kinase, and provides new insight into how Hh morphogen progressively activates Smo.

  7. Regulation of Smoothened Phosphorylation and High-Level Hedgehog Signaling Activity by a Plasma Membrane Associated Kinase.

    Science.gov (United States)

    Li, Shuangxi; Li, Shuang; Han, Yuhong; Tong, Chao; Wang, Bing; Chen, Yongbin; Jiang, Jin

    2016-06-01

    Hedgehog (Hh) signaling controls embryonic development and adult tissue homeostasis through the G protein coupled receptor (GPCR)-family protein Smoothened (Smo). Upon stimulation, Smo accumulates on the cell surface in Drosophila or primary cilia in vertebrates, which is thought to be essential for its activation and function, but the underlying mechanisms remain poorly understood. Here we show that Hh stimulates the binding of Smo to a plasma membrane-associated kinase Gilgamesh (Gish)/CK1γ and that Gish fine-tunes Hh pathway activity by phosphorylating a Ser/Thr cluster (CL-II) in the juxtamembrane region of Smo carboxyl-terminal intracellular tail (C-tail). We find that CL-II phosphorylation is promoted by protein kinase A (PKA)-mediated phosphorylation of Smo C-tail and depends on cell surface localization of both Gish and Smo. Consistent with CL-II being critical for high-threshold Hh target gene expression, its phosphorylation appears to require higher levels of Hh or longer exposure to the same level of Hh than PKA-site phosphorylation on Smo. Furthermore, we find that vertebrate CK1γ is localized at the primary cilium to promote Smo phosphorylation and Sonic hedgehog (Shh) pathway activation. Our study reveals a conserved mechanism whereby Hh induces a change in Smo subcellular localization to promote its association with and activation by a plasma membrane localized kinase, and provides new insight into how Hh morphogen progressively activates Smo.

  8. Regulation of Smoothened Phosphorylation and High-Level Hedgehog Signaling Activity by a Plasma Membrane Associated Kinase

    Science.gov (United States)

    Tong, Chao; Wang, Bing; Chen, Yongbin; Jiang, Jin

    2016-01-01

    Hedgehog (Hh) signaling controls embryonic development and adult tissue homeostasis through the G protein coupled receptor (GPCR)-family protein Smoothened (Smo). Upon stimulation, Smo accumulates on the cell surface in Drosophila or primary cilia in vertebrates, which is thought to be essential for its activation and function, but the underlying mechanisms remain poorly understood. Here we show that Hh stimulates the binding of Smo to a plasma membrane-associated kinase Gilgamesh (Gish)/CK1γ and that Gish fine-tunes Hh pathway activity by phosphorylating a Ser/Thr cluster (CL-II) in the juxtamembrane region of Smo carboxyl-terminal intracellular tail (C-tail). We find that CL-II phosphorylation is promoted by protein kinase A (PKA)-mediated phosphorylation of Smo C-tail and depends on cell surface localization of both Gish and Smo. Consistent with CL-II being critical for high-threshold Hh target gene expression, its phosphorylation appears to require higher levels of Hh or longer exposure to the same level of Hh than PKA-site phosphorylation on Smo. Furthermore, we find that vertebrate CK1γ is localized at the primary cilium to promote Smo phosphorylation and Sonic hedgehog (Shh) pathway activation. Our study reveals a conserved mechanism whereby Hh induces a change in Smo subcellular localization to promote its association with and activation by a plasma membrane localized kinase, and provides new insight into how Hh morphogen progressively activates Smo. PMID:27280464

  9. Potentiating activity of luteolin on membrane permeabilizing agent and ATPase inhibitor against methicillin-resistant Staphylococcus aureus.

    Science.gov (United States)

    Joung, Dae-Ki; Lee, Young-Seob; Han, Sin-Hee; Lee, Sang-Won; Cha, Seon-Woo; Mun, Su-Hyun; Kong, Ryong; Kang, Ok-Hwa; Song, Ho-Jun; Shin, Dong-Won; Kwon, Dong-Yeul

    2016-01-01

    To investigate the mechanism of antibacterial activity of luteolin (LUT) against methicillin-resistant Staphylococcus aureus (MRSA). The mechanism of anti-MRSA activity of LUT was analyzed by the viability assay in membrane permeabilizing agent, ATPase inhibitors, and peptidoglycan (PGN) derived from Staphylococcus aureus (S. aureus). Also, transmission electron microscopy was used to monitor survival characteristics and changes in S. aureus morphology. Compared to the LUT alone, the optical density of suspensions treated with the combination of 125 μg/mL Tris and 250 μg/mL N,N'-dicyclohexylcarbodiimide were reduced to 60% and 46% of the control, respectively. PGN (15.6 μg/mL) gradually impeded the activity of LUT, and PGN (62.5 μg/mL) completely blocked the activity of LUT on S. aureus. Increased susceptibility to LUT with the Tris-dicyclohexylcarbodiimide combinations is evident in all tested MRSA isolates. The results indicate LUT synergy in increasing cytoplasmic membrane permeability and inhibiting ATPase. S. aureus PGN directly blocks the antibacterial activity of LUT, suggesting the direct binding of LUT with PGN. These findings may be validated for the development of antibacterial agent for low MRSA resistance. Copyright © 2016 Hainan Medical College. Production and hosting by Elsevier B.V. All rights reserved.

  10. Activity ranking of synthetic analogs targeting vascular endothelial growth factor receptor 2 by an integrated cell membrane chromatography system.

    Science.gov (United States)

    Wang, Dongyao; Lv, Diya; Chen, Xiaofei; Liu, Yue; Ding, Xuan; Jia, Dan; Chen, Langdong; Zhu, Zhenyu; Cao, Yan; Chai, Yifeng

    2015-12-01

    Evaluating the biological activities of small molecules represents an important part of the drug discovery process. Cell membrane chromatography (CMC) is a well-developed biological chromatographic technique. In this study, we have developed combined SMMC-7721/CMC and HepG2/CMC with high-performance liquid chromatography and time-of-flight mass spectrometry to establish an integrated screening platform. These systems was subsequently validated and used for evaluating the activity of quinazoline compounds, which were designed and synthesized to target vascular endothelial growth factor receptor 2. The inhibitory activities of these compounds towards this receptor were also tested using a classical caliper mobility shift assay. The results revealed a significant correlation between these two methods (R(2) = 0.9565 or 0.9420) for evaluating the activities of these compounds. Compared with traditional methods of evaluating the activities analogous compounds, this integrated cell membrane chromatography screening system took less time and was more cost effective, indicating that it could be used as a practical method in drug discovery. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Plasma membrane ATPases

    DEFF Research Database (Denmark)

    Palmgren, Michael Broberg; Bækgaard, Lone; Lopez Marques, Rosa Laura

    2011-01-01

    membrane include ABC transporters, vacuolar (V-type) H+ pumps, and P-type pumps. These pumps all utilize ATP as a fuel for energizing pumping. This review focuses on the physiological roles of plasma membrane P-type pumps, as they represent the major ATP hydrolytic activity in this membrane....

  12. Potent synergistic in vitro interaction between nonantimicrobial membrane-active compounds and itraconazole against clinical isolates of Aspergillus fumigatus resistant to itraconazole.

    NARCIS (Netherlands)

    Afeltra, J.; Vitale, R.G.; Mouton, J.W.; Verweij, P.E.

    2004-01-01

    To develop new approaches for the treatment of invasive infections caused by Aspergillus fumigatus, the in vitro interactions between itraconazole (ITZ) and seven different nonantimicrobial membrane-active compounds--amiodarone (AMD), amiloride, lidocaine, lansoprazole (LAN), nifedipine (NIF),

  13. Activation of AMPK in human fetal membranes alleviates infection-induced expression of pro-inflammatory and pro-labour mediators.

    Science.gov (United States)

    Lim, R; Barker, G; Lappas, M

    2015-04-01

    In non-gestational tissues, the activation of adenosine monophosphate (AMP)-activated kinase (AMPK) is associated with potent anti-inflammatory actions. Infection and/or inflammation, by stimulating pro-inflammatory cytokines and matrix metalloproteinase (MMP)-9, play a central role in the rupture of fetal membranes. However, no studies have examined the role of AMPK in human labour. Fetal membranes, from term and preterm, were obtained from non-labouring and labouring women, and after preterm pre-labour rupture of membranes (PPROM). AMPK activity was assessed by Western blotting of phosphorylated AMPK expression. To determine the effect of AMPK activators on pro-inflammatory cytokines, fetal membranes were pre-treated with AMPK activators then stimulated with bacterial products LPS and flagellin or viral dsDNA analogue poly(I:C). Primary amnion cells were used to determine the effect of AMPK activators on IL-1β-stimulated MMP-9 expression. AMPK activity was decreased with term labour. There was no effect of preterm labour. AMPK activity was also decreased in preterm fetal membranes, in the absence of labour, with PROM compared to intact membranes. AMPK activators AICAR, phenformin and A769662 significantly decreased IL-6 and IL-8 stimulated by LPS, flagellin and poly(I:C). Primary amnion cells treated with AMPK activators significantly decreased IL-1β-induced MMP-9 expression. The decrease in AMPK activity in fetal membranes after spontaneous term labour and PPROM indicates an anti-inflammatory role for AMPK in human labour and delivery. The use of AMPK activators as possible therapeutics for threatened preterm labour would be an exciting future avenue of research. Copyright © 2015 Elsevier Ltd. All rights reserved.

  14. Neither hollow-fibre membrane filters nor activated-charcoal filters remove fluoride from fluoridated tap water.

    Science.gov (United States)

    Konno, Hideki; Yaegaki, Ken; Tanaka, Tomoko; Sato, Tsutomu; Itai, Kazuyoshi; Imai, Toshio; Murata, Takatoshi; Herai, Mayumi

    2008-06-01

    Previous reports of the reduction of fluoride concentrations in fluoridated water by domestic water treatment systems have indicated that further supplementation with fluoride is required. However, the absorption of fluoride by filters has not yet been directly identified. If these filters do not absorb fluoride, further fluoride supplementation may increase fluorosis. In this study, we determined whether filtering systems absorb fluoride ions. We directly measured the amounts of fluoride absorbed by activated-carbon filters or hollow-fibre membrane filters using pyrohydrolysis of the filters and flow-injection analysis, the sensitivity of which is more than 100 times greater than that of conventional methods. We made fluoride solutions of pure or tap water and determined changes in fluoride concentration as a result of filtering with a fluoride electrode. Hollow-fibre membrane filters did not affect fluoride concentrations in the fluoridated water, but activated-carbon filters removed some fluoride, especially from the pure-water solution. Filtering a pure-water solution with a fluoride concentration of 0.8 mg F/L reduced the fluoride concentration until 210 L of the solution had been filtered. However, filtering a tap-water solution of 0.8 mg F/L reduced the fluoride concentration only until 8 L had been filtered. The concentration of absorbed fluoride in the filter at 10 L of filtration was 4.7 mg/kg activated carbon. Further fluoride supplementation of fluoridated water should not be necessary, regardless of whether an activated-carbon or hollow-fibre membrane filter is installed on a domestic water treatment system.

  15. Athermal alterations in the structure in the canalicular membrane and ATPase activity induced by thermal levels of microwave radiation

    International Nuclear Information System (INIS)

    Phelan, A.M.; Neubauer, C.F.; Timm, R.; Neirenberg, J.; Lange, D.G.

    1994-01-01

    Sprague-Dawley rats (200-250 g) were exposed 30 min/day for 4 days to thermogenic levels (rectal temperature increase of 2.2 degrees C) of microwave radiation [2.45 GHz, 80 mW/cm 2 , continuous-wave mode (CW)] or to a radiant heat source resulting in an equivalent increase in body temperature of 2.2 degrees C. On the fifth day the animals were sacrificed and their livers removed. The canalicular membranes were isolated and evaluated for adenosinetriphosphatase (ATPase) activity, total fatty acid composition and membrane fluidity characteristics. Mg ++ -ATPase activity (V max ) decreased by 48.5% in the group exposed to microwave radiation, with no significant change in the group exposed to radiant heat. The decrease in Mg ++ -ATPase was partially compensated by a concomitant increase in Na + /K + -ATPase activity (170% increase in V max over control) in animals exposed to microwave radiation, while no change occurred in the group exposed to radiant heat. This alteration in ATPase activity in the group exposed to microwave radiation is associated with a large decrease in the ratio of saturated to unsaturated fatty acids. Conversely, the group exposed to radiant heat had an increase in the ratio of saturated to unsaturated fatty acids. The most dramatic changes were found in the levels of arachidonic acid. Finally, the electron paramagnetic resonance (EPR) spin label technique used to measure the fluidity of the canalicular membranes of the animals in the three groups (sham, microwave radiation and radiant heat) indicated that the results were different in the three groups, reflecting the changes found in their fatty acid composition. The physiological response to open-quotes equivalentclose quotes thermal loads in rats is expressed differently for different types of energy sources. Possible mechanisms producing these divergent thermogenic responses are discussed. 34 refs., 3 figs., 2 tabs

  16. Membrane phospholipid augments cytochrome P4501a enzymatic activity by modulating structural conformation during detoxification of xenobiotics.

    Directory of Open Access Journals (Sweden)

    Manik C Ghosh

    Full Text Available Cytochrome P450 is a superfamily of membrane-bound hemoprotein that gets involved with the degradation of xenobiotics and internal metabolites. Accumulated body of evidence indicates that phospholipids play a crucial role in determining the enzymatic activity of cytochrome P450 in the microenvironment by modulating its structure during detoxification; however, the structure-function relationship of cytochrome P4501A, a family of enzymes responsible for degrading lipophilic aromatic hydrocarbons, is still not well defined. Inducibility of cytochrome P4501A in cultured catfish hepatocytes in response to carbofuran, a widely used pesticide around the world, was studied earlier in our laboratory. In this present investigation, we observed that treating catfish with carbofuran augmented total phospholipid in the liver. We examined the role of phospholipid on the of cytochrome P4501A-marker enzyme which is known as ethoxyresorufin-O-deethylase (EROD in the context of structure and function. We purified the carbofuran-induced cytochrome P4501A protein from catfish liver. Subsequently, we examined the enzymatic activity of purified P4501A protein in the presence of phospholipid, and studied how the structure of purified protein was influenced in the phospholipid environment. Membrane phospholipid appeared to accelerate the enzymatic activity of EROD by changing its structural conformation and thus controlling the detoxification of xenobiotics. Our study revealed the missing link of how the cytochrome P450 restores its enzymatic activity by changing its structural conformation in the phospholipid microenvironment.

  17. Definition of the "Drug-Angiogenic-Activity-Index" that allows the quantification of the positive and negative angiogenic active drugs: a study based on the chorioallantoic membrane model.

    Science.gov (United States)

    Demir, Resit; Peros, Georgios; Hohenberger, Werner

    2011-06-01

    Since the introduction of the angiogenic therapy by Folkman et al. in the 1970'ies many antiangiogenic drugs were identified. Only few of them are still now in clinical use. Also the Vascular Endothelial Growth Factor (VEGF), the cytokine with the highest angiogenic activity, has been identified. Its antagonist, Bevacizumab, is produced and admitted for the angiogenic therapy in first line for metastatic colorectal cancer. When we look at preclinical studies, they fail of in vivo models that define the "Drug-Angiogenic-Activity-Index" of angiogenic or antiangiogenic drugs. This work proposes a possible standardized procedure to define the "Drug Angiogenic Activity Index" by counting the vascular intersections (VIS) on the Chorioallantoic Membrane after drug application. The equation was defined as follows: {ΔVIS[Drug]-ΔVIS[Control]} / Δ VIS[Control]. For VEGF a Drug-Angiogenic-Activity-Index of 0.92 was found and for Bevacizumab a -1. This means almost that double of the naturally angiogenic activity was achieved by VEGF on the Chorioallantoic membrane. A complete blocking of naturally angiogenic activity was observed after Bevacizumabs application. Establishing the "Drug-Angiogenic-Activity-Index" in the preclinical phase will give us an impact of effectiveness for the new constructed antiangiogenic drugs like the impact of effectiveness in the cortisone family.

  18. Therapeutic drug monitoring of flucytosine in serum using a SERS-active membrane system

    Science.gov (United States)

    Berger, Adam G.; White, Ian M.

    2017-02-01

    A need exists for near real-time therapeutic drug monitoring (TDM), in particular for antibiotics and antifungals in patient samples at the point-of-care. To truly fit the point-of-care need, techniques must be rapid and easy to use. Here we report a membrane system utilizing inkjet-fabricated surface enhanced Raman spectroscopy (SERS) sensors that allows sensitive and specific analysis despite the elimination of sophisticated chromatography equipment, expensive analytical instruments, and other systems relegated to the central lab. We utilize inkjet-fabricated paper SERS sensors as substrates for 5FC detection; the use of paper-based SERS substrates leverages the natural wicking ability and filtering properties of microporous membranes. We investigate the use of microporous membranes in the vertical flow assay to allow separation of the flucytosine from whole blood. The passive vertical flow assay serves as a valuable method for physical separation of target analytes from complex biological matrices. This work further establishes a platform for easy, sensitive, and specific TDM of 5FC from whole blood.

  19. Antibacterial activity of ruthenium(II) polypyridyl complex manipulated by membrane permeability and cell morphology.

    Science.gov (United States)

    Sun, Dongdong; Zhang, Weiwei; Lv, Mengting; Yang, Endong; Zhao, Qian; Wang, Weiyun

    2015-01-01

    This study investigates the antibacterial effects of the ruthenium(II) complex RuBP and the mechanism of RuBP action on bacteria. Results show that RuBP can inhibit the growth of Gram-positive bacteria, such as Staphylococcus aureus and Micrococcus tetragenus. Cellular uptake and laser confocal microscopic studies reveal the efficient uptake of RuBP by M. tetragenus cells. Scanning electron microscopic observations of the morphologies of M. tetragenus and S. aureus treated with RuBP further confirm that direct contact of both bacteria with RuBP can damage the cell membrane and membrane integrity, which may eventually induce growth inhibition and bacterial death. After RuBP treatment, the electrical conductivity of the bacterial suspensions increases. Spectroscopic studies and agarose gel electrophoresis indicate that intact DNA and RNA decrease or disappear in RuBP-treated bacterial cells, thus demonstrating that RuBP performs its antibacterial function by increasing the permeability of cell membranes. This study provides new insights for understanding the antibacterial actions of RuBP and designing metal complex antibiotics for other biomedical applications. Copyright © 2015 Elsevier Ltd. All rights reserved.

  20. Autographa californica multiple nucleopolyhedrovirus GP64 protein: Analysis of domain I and V amino acid interactions and membrane fusion activity

    Energy Technology Data Exchange (ETDEWEB)

    Yu, Qianlong [State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Northwest Loess Plateau Crop Pest Management of Ministry of Agriculture, College of Plant Protection, Northwest A& F University, Yangling, Shaanxi 712100 (China); Blissard, Gary W. [Boyce Thompson Institute, Cornell University, Ithaca, NY 14853, United State (United States); Liu, Tong-Xian [State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Northwest Loess Plateau Crop Pest Management of Ministry of Agriculture, College of Plant Protection, Northwest A& F University, Yangling, Shaanxi 712100 (China); Li, Zhaofei, E-mail: zhaofeili73@outlook.com [State Key Laboratory of Crop Stress Biology for Arid Areas, Key Laboratory of Northwest Loess Plateau Crop Pest Management of Ministry of Agriculture, College of Plant Protection, Northwest A& F University, Yangling, Shaanxi 712100 (China)

    2016-01-15

    The Autographa californica multiple nucleopolyhedrovirus GP64 is a class III viral fusion protein. Although the post-fusion structure of GP64 has been solved, its pre-fusion structure and the detailed mechanism of conformational change are unknown. In GP64, domain V is predicted to interact with two domain I segments that flank fusion loop 2. To evaluate the significance of the amino acids involved in these interactions, we examined 24 amino acid positions that represent interacting and conserved residues within domains I and V. In several cases, substitution of a single amino acid involved in a predicted interaction disrupted membrane fusion activity, but no single amino acid pair appears to be absolutely required. We identified 4 critical residues in domain V (G438, W439, T452, and T456) that are important for membrane fusion, and two residues (G438 and W439) that appear to be important for formation or stability of the pre-fusion conformation of GP64. - Highlights: • The baculovirus envelope glycoprotein GP64 is a class III viral fusion protein. • The detailed mechanism of conformational change of GP64 is unknown. • We analyzed 24 positions that might stabilize the post-fusion structure of GP64. • We identified 4 residues in domain V that were critical for membrane fusion. • Two residues are critical for formation of the pre-fusion conformation of GP64.

  1. Autographa californica multiple nucleopolyhedrovirus GP64 protein: Analysis of domain I and V amino acid interactions and membrane fusion activity

    International Nuclear Information System (INIS)

    Yu, Qianlong; Blissard, Gary W.; Liu, Tong-Xian; Li, Zhaofei

    2016-01-01

    The Autographa californica multiple nucleopolyhedrovirus GP64 is a class III viral fusion protein. Although the post-fusion structure of GP64 has been solved, its pre-fusion structure and the detailed mechanism of conformational change are unknown. In GP64, domain V is predicted to interact with two domain I segments that flank fusion loop 2. To evaluate the significance of the amino acids involved in these interactions, we examined 24 amino acid positions that represent interacting and conserved residues within domains I and V. In several cases, substitution of a single amino acid involved in a predicted interaction disrupted membrane fusion activity, but no single amino acid pair appears to be absolutely required. We identified 4 critical residues in domain V (G438, W439, T452, and T456) that are important for membrane fusion, and two residues (G438 and W439) that appear to be important for formation or stability of the pre-fusion conformation of GP64. - Highlights: • The baculovirus envelope glycoprotein GP64 is a class III viral fusion protein. • The detailed mechanism of conformational change of GP64 is unknown. • We analyzed 24 positions that might stabilize the post-fusion structure of GP64. • We identified 4 residues in domain V that were critical for membrane fusion. • Two residues are critical for formation of the pre-fusion conformation of GP64.

  2. Stabilization of Erythrocyte Membranes by Polyamines

    Science.gov (United States)

    Ballas, Samir K.; Mohandas, Narla; Marton, Laurence J.; Shohet, Stephen B.

    1983-04-01

    Using a laser diffraction technique, we have studied the effects of putrescine, spermidine, and spermine, the three physiologic polyamines, on the deformability and mechanical stability of human erythrocyte membranes. Ghosts resealed with polyamines were subjected to high fluid shear stress in an ektacytometer. All polyamines increased the membrane shear modulus (decreased deformability) in a concentration- and time-dependent manner. The order of effectiveness was spermine > spermidine > putrescine. At 10 μ M, spermine appreciably decreased membrane deformability. For the measurement of membrane mechanical stability, resealed ghosts were subjected to constant high shear stress in the ektacytometer and deformability was continuously recorded as the deformable ghosts fragmented into rigid spherical vesicles. Polyamines, especially spermine, caused a noticeable increase in the t1/2 for fragmentation. These effects could not be ascribed to proteolysis or Ca2+-induced transglutamination. That the effects of polyamines were specific and not simply due to their positive charge was demonstrated by the finding that Ca2+ and Mg2+ destabilized the erythrocyte membrane as evidenced by decreasing the t1/2 for fragmentation. Extracellular polyamines were not effective except under conditions that caused significant accumulation inside the cell. The data indicate that intracellular physiologic polyamines, especially spermine, decrease erythrocyte membrane deformability and stabilize the membrane skeleton, making it more resistant to fragmentation.

  3. Using fluorine nuclear magnetic resonance to probe the interaction of membrane-active peptides with the lipid bilayer.

    Science.gov (United States)

    Buer, Benjamin C; Chugh, Jeetender; Al-Hashimi, Hashim M; Marsh, E Neil G

    2010-07-13

    A variety of biologically active peptides exert their function through direct interactions with the lipid membrane of the cell. These surface interactions are generally transient and highly dynamic, making them hard to study. Here we have examined the feasibility of using solution phase (19)F nuclear magnetic resonance (NMR) to study peptide-membrane interactions. Using the antimicrobial peptide MSI-78 as a model system, we demonstrate that peptide binding to either small unilamellar vesicles (SUVs) or bicelles can readily be detected by simple one-dimensional (19)F NMR experiments with peptides labeled with l-4,4,4-trifluoroethylglycine. The (19)F chemical shift associated with the peptide-membrane complex is sensitive both to the position of the trifluoromethyl reporter group (whether in the hydrophobic face or positively charged face of the amphipathic peptide) and to the curvature of the lipid bilayer (whether the peptide is bound to SUVs or bicelles). (19)F spin echo experiments using the Carr-Purcell-Meiboom-Gill pulse sequence were used to measure the transverse relaxation (T(2)) of the nucleus and thereby examine the local mobility of the MSI-78 analogues bound to bicelles. The fluorine probe positioned in the hydrophobic face of the peptide relaxes at a rate that correlates with the tumbling of the bicelle, suggesting that it is relatively immobile, whereas the probe at the positively charged face relaxes more slowly, indicating this position is much more dynamic. These results are in accord with structural models of MSI-78 bound to lipids and point to the feasibility of using fluorine-labeled peptides to monitor peptide-membrane interactions in living cells.

  4. Antiparasitic activity and effect of casearins isolated from Casearia sylvestris on Leishmania and Trypanosoma cruzi plasma membrane.

    Science.gov (United States)

    Bou, Diego Dinis; Tempone, André G; Pinto, Érika G; Lago, João Henrique G; Sartorelli, Patricia

    2014-04-15

    Leishmaniasis and Chagas disease are infectious diseases caused by parasite Leishmania sp. and Trypanosoma cruzi, respectively, and are included among the most neglected diseases in several underdeveloped and developing countries, with an urgent demand for new drugs. Considering the antiparasitic potential of MeOH extract from leaves of Casearia sylvestris Sw. (Salicaceae), a bioguided fractionation was conducted and afforded four active clerodane diterpenes (casearins A, B, G, and J). The obtained results indicated a superior efficacy of tested casearins against trypomastigotes of T. cruzi, with IC50 values ranging from 0.53 to 2.77 μg/ml. Leishmania infantum promastigotes were also susceptible to casearins, with IC50 values in a range between 4.45 and 9.48 μg/ml. These substances were also evaluated for mammalian cytotoxicity against NCTC cells resulting in 50% cytotoxic concentrations (CC50) ranging from 1.46 to 13.76 μg/ml. Additionally, the action of casearins on parasite membranes was investigated using the fluorescent probe SYTOX Green. The obtained results demonstrated a strong interaction of casearins A and B to the plasma membrane of T. cruzi parasites, corroborating their higher efficacy against these parasites. In contrast, the tested casearins induced no alteration in the permeability of plasma membrane of Leishmania parasites, suggesting that biochemical differences between Leishmania and T. cruzi plasma membrane might have contributed to the target effect of casearins on trypomastigotes. Thus, considering the importance of studying novel and selective drug candidates against protozoans, casearins A, B, G, and J could be used as tools to future drug design studies. Copyright © 2014 Elsevier GmbH. All rights reserved.

  5. Wind sock deformity in rectal atresia

    International Nuclear Information System (INIS)

    Hosseini, Seyed M V; Ghahramani, Farhad; Shamsaeefar, Alireza; Razmi, Tannaz; Zarenezhad, Mohammad

    2009-01-01

    Rectal atresia is a rare anorectal deformity. It usually presents with neonatal obstruction and it is often a complete membrane or severe stenosis. Windsock deformity has not been reported in rectal atresia especially, having been missed for 2 years. A 2-year-old girl reported only a severe constipation despite having a 1.5-cm anal canal in rectal examination with scanty discharge. She underwent loop colostomy and loopogram, which showed a wind sock deformity of rectum with mega colon. The patient underwent abdominoperineal pull-through with good result and follow-up. This is the first case of the wind sock deformity in rectal atresia being reported after 2 years of age. (author)

  6. Deformation and recrystallization mechanisms in actively extruding salt fountain: Microstructural evidence for a switch in deformation mechanisms with increased availability of meteoric water and decreased grain size (Qum Kuh, central Iran)

    Czech Academy of Sciences Publication Activity Database

    Desbois, G.; Závada, Prokop; Schléder, Z.; Urai, J. L.

    2010-01-01

    Roč. 32, č. 4 (2010), s. 580-594 ISSN 0191-8141 Grant - others:Deutsche Forschungsgemeinschaft(DE) UR 64/9-2 Institutional research plan: CEZ:AV0Z30120515 Keywords : rocksalt * salt extrusion * gamma - irradiation * deformation mechanisms * microstructure Subject RIV: DB - Geology ; Mineralogy Impact factor: 1.911, year: 2010

  7. 3D geodetic monitoring slope deformations

    Directory of Open Access Journals (Sweden)

    Weiss Gabriel

    1996-06-01

    Full Text Available For plenty of slope failures that can be found in Slovakia is necessary and very important their geodetic monitoring (because of their activity, reactivisations, checks. The paper gives new methodologies for these works, using 3D terrestrial survey technologies for measurements in convenient deformation networks. The design of an optimal type of deformation model for various kinds of landslides and their exact processing with an efficient testing procedure to determine the kinematics of the slope deformations are presented too.

  8. Moist and soft, dry and stiff: a review of neutron experiments on hydration-dynamics-activity relations in the purple membrane of Halobacterium salinarum.

    Science.gov (United States)

    Zaccai, G

    2000-08-30

    Twenty-five years of neutron experiments on hydration and thermal dynamics in purple membranes of Halobacterium salinarum are reviewed. Neutron diffraction, elastic and quasielastic scattering, allowed to map the distribution of water and lipids and to measure thermal fluctuations and correlation times in the membranes, under various conditions of temperature, hydration and lipid environment. Strong correlations were established between dynamics parameters and the activity of bacteriorhodopsin (the purple membrane protein), as a light driven proton pump supporting the hypothesis that the influence of hydration on activity is in fact due to its effects on membrane thermal dynamics. Hydrogen-deuterium labelling experiments highlighted stiffer and softer parts in the bacteriorhodopsin structure. The soft parts would allow the conformational changes involved in activity, while the stiffer ones may control a valve-like function in vectorial proton transfer.

  9. Alteration in membrane fluidity and lipid composition, and modulation of H(+)-ATPase activity in Saccharomyces cerevisiae caused by decanoic acid.

    Science.gov (United States)

    Alexandre, H; Mathieu, B; Charpentier, C

    1996-03-01

    Decanoic acid, a lipophilic agent, inhibited in vitro the plasma membrane H(+)-ATPase of Saccharomyces cerevisiae grown in YPD medium. Conversely, when decanoic acid (35 microM) was present in the growth medium, the measured H(+)-ATPase activity was four times higher than that of control cells. Km, and pH and orthovanadate sensitivity were the same for the two growth conditions, which indicated that H(+)-ATPase activation was not due to conformational changes in the enzyme. The activation process was not entirely reversible which showed that plasma membrane H(+)-ATPase activation is due to several mechanisms. 1,6-diphenyl-1,3,5-hexatriene anisotropy performed on protoplasts from cells grown in YPD revealed that as decanoic acid concentration was increased, anisotropy significantly decreased, i.e. membrance fluidity increased. Cells grown in media containing decanoic acid exhibited greater membrane fluidity compared with control cells. Furthermore, these cells did not show any fluidifying effect when increased concentrations of decanoic acid were added. Chemical analysis of cell membrane lipid composition revealed a modification in the distribution of the phospholipid fatty acids and sterols in cells grown in the presence of 35 microM decanoic acid compared with control cells. Our results support the view that the plasma membrane H(+)-ATPase activation induced by decanoic acid is correlated with an alteration in membrane lipid constituents.

  10. Discovery of membrane active benzimidazole quinolones-based topoisomerase inhibitors as potential DNA-binding antimicrobial agents.

    Science.gov (United States)

    Zhang, Ling; Addla, Dinesh; Ponmani, Jeyakkumar; Wang, Ao; Xie, Dan; Wang, Ya-Nan; Zhang, Shao-Lin; Geng, Rong-Xia; Cai, Gui-Xin; Li, Shuo; Zhou, Cheng-He

    2016-03-23

    A series of novel benzimidazole quinolones as potential antimicrobial agents were designed and synthesized. Most of the prepared compounds exhibited good or even stronger antimicrobial activities in comparison with reference drugs. The most potent compound 15m was membrane active and did not trigger the development of resistance in bacteria. It not only inhibited the formation of biofilms but also disrupted the established Staphylococcus aureus and Escherichia coli biofilms. It was able to inhibit the relaxation activity of E. coli topoisomerase IV at 10 μM concentration. Moreover, this compound also showed low toxicity against mammalian cells. Molecular modeling and experimental investigation of compound 15m with DNA suggested that this compound could effectively bind with DNA to form a steady 15m-DNA complex which might further block DNA replication to exert the powerful bioactivities. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

  11. Membrane-Cytoskeleton Dynamics in the Course of Mast Cell Activation

    Czech Academy of Sciences Publication Activity Database

    Dráber, Pavel; Dráber, Petr

    2015-01-01

    Roč. 1220, č. 1 (2015), s. 219-237 ISSN 1064-3745 R&D Projects: GA ČR GA204/09/1777; GA ČR(CZ) GA14-09807S; GA ČR(CZ) GA14-00703S; GA ČR(CZ) GBP302/12/G101; GA MŠk LC545; GA MŠk LD12073; GA MŠk(CZ) LD13015; GA AV ČR KAN200520701; GA AV ČR(CZ) M200520901 Institutional support: RVO:68378050 Keywords : mast cell s * plasma membrane * actin Subject RIV: EB - Genetics ; Molecular Biology

  12. Segments Crucial for Membrane Translocation and Pore-forming Activity of Bordetella Adenylate Cyclase Toxin

    Czech Academy of Sciences Publication Activity Database

    Basler, Marek; Knapp, O.; Mašín, Jiří; Fišer, R.; Maier, E.; Benz, R.; Šebo, Peter; Osička, Radim

    2007-01-01

    Roč. 282, č. 17 (2007), s. 12419-12429 ISSN 0021-9258 R&D Projects: GA MŠk 1M0506; GA AV ČR IAA5020406 Grant - others:XE(XE) European Union 6th FP contract LSHB-CT-2003-503582 THERAVAC Institutional research plan: CEZ:AV0Z50200510 Source of funding: R - rámcový projekt EK Keywords : bordetella * adenylate cyclase toxin * ac membrane translocation Subject RIV: EE - Microbiology, Virology Impact factor: 5.581, year: 2007

  13. Active Probes for Imaging Membrane Dynamics of Live Cells with High Spatial and Temporal Resolution over Extended Time Scales and Areas.

    Science.gov (United States)

    Wang, Huaimin; Feng, Zhaoqianqi; Del Signore, Steven J; Rodal, Avital A; Xu, Bing

    2018-03-14

    Despite the advancement of molecular imaging techniques, there is an unmet need for probes for direct imaging of membrane dynamics of live cells. Here we report a novel type of active (or enzyme responsive) probes to directly image membrane dynamics of live cells with high spatial and temporal resolution over extended time scales and areas. Because lipid rafts enrich cholesterols and GPI-anchored enzymes (e.g., ectophosphatases), we design probes that consist of an enzymatic trigger, a fluorophore, and a cholesterol that are affinitive to the cell membrane. Being water-soluble and as the substrate of ectophosphatase, these cell compatible probes preferentially and rapidly assemble in plasma membrane, exhibit strong fluorescence, work at micromolar concentrations, and easily achieve high resolution monitoring of nanoscale heterogeneity in membranes of live cells, the release of exosomes, and the membrane dynamics of live cells. This work provides a facile means to link membrane dynamics and heterogeneity to cellular processes for understanding the interactions between membranes and proteins.

  14. Synergistic activation of protein kinase Calpha, -betaI, and -gamma isoforms induced by diacylglycerol and phorbol ester: roles of membrane association and activating conformational changes.

    Science.gov (United States)

    Slater, S J; Milano, S K; Stagliano, B A; Gergich, K J; Ho, C; Mazurek, A; Taddeo, F J; Kelly, M B; Yeager, M D; Stubbs, C D

    1999-03-23

    Protein kinase Calpha (PKCalpha) has been shown to contain two discrete activator sites with differing binding affinities for phorbol esters and diacylglycerols. The interaction of diacylglycerol with a low-affinity phorbol ester binding site leads to enhanced high-affinity phorbol ester binding and to a potentiated level of activity [Slater, S. J., Ho, C., Kelly, M. B., Larkin, J. D. , Taddeo, F. J., Yeager, M. D., and Stubbs, C. D. (1996) J. Biol. Chem. 271, 4627-4631]. In this study, the mechanism of this enhancement of activity was examined with respect to the Ca2+ dependences of membrane association and accompanying conformational changes that lead to activation. The asso