Biliary Secretion of Quasi-Enveloped Human Hepatitis A Virus

PubMed Central

Hirai-Yuki, Asuka; Hensley, Lucinda; Whitmire, Jason K.

2016-01-01

ABSTRACT Hepatitis A virus (HAV) is an unusual picornavirus that is released from cells cloaked in host-derived membranes. These quasi-enveloped virions (eHAV) are the only particle type circulating in blood during infection, whereas only nonenveloped virions are shed in feces. The reason for this is uncertain. Hepatocytes, the only cell type known to support HAV replication in vivo, are highly polarized epithelial cells with basolateral membranes facing onto hepatic (blood) sinusoids and apical membranes abutting biliary canaliculi from which bile is secreted to the gut. To assess whether eHAV and nonenveloped virus egress from cells via vectorially distinct pathways, we studied infected polarized cultures of Caco-2 and HepG2-N6 cells. Most (>99%) progeny virions were released apically from Caco-2 cells, whereas basolateral (64%) versus apical (36%) release was more balanced with HepG2-N6 cells. Both apically and basolaterally released virions were predominantly enveloped, with no suggestion of differential vectorial release of eHAV versus naked virions. Basolateral to apical transcytosis of either particle type was minimal (<0.02%/h) in HepG2-N6 cells, arguing against this as a mechanism for differences in membrane envelopment of serum versus fecal virus. High concentrations of human bile acids converted eHAV to nonenveloped virions, whereas virus present in bile from HAV-infected Ifnar1−/− Ifngr1−/− and Mavs−/− mice banded over a range of densities extending from that of eHAV to that of nonenveloped virions. We conclude that nonenveloped virions shed in feces are derived from eHAV released across the canalicular membrane and stripped of membranes by the detergent action of bile acids within the proximal biliary canaliculus. PMID:27923925

Time-Resolved Neutron Interferometry and the Mechanism of Electromechanical Coupling in Voltage-Gated Ion Channels.

PubMed

Blasie, J Kent

2018-01-01

The mechanism of electromechanical coupling for voltage-gated ion channels (VGICs) involved in neurological signal transmission, primarily Nav- and Kv-channels, remains unresolved. Anesthetics have been shown to directly impact this mechanism, at least for Kv-channels. Molecular dynamics computer simulations can now predict the structures of VGICs embedded within a hydrated phospholipid bilayer membrane as a function of the applied transmembrane voltage, but significant assumptions are still necessary. Nevertheless, these simulations are providing new insights into the mechanism of electromechanical coupling at the atomic level in 3-D. We show that time-resolved neutron interferometry can be used to investigate directly the profile structure of a VGIC, vectorially oriented within a single hydrated phospholipid bilayer membrane at the solid-liquid interface, as a function of the applied transmembrane voltage in the absence of any assumptions or potentially perturbing modifications of the VGIC protein and/or the host membrane. The profile structure is a projection of the membrane's 3-D structure onto the membrane normal and, in the absence of site-directed deuterium labeling, is provided at substantially lower spatial resolution than the atomic level. Nevertheless, this novel approach can be used to directly test the validity of the predictions from molecular dynamics simulations. We describe the key elements of our novel experimental approach, including why each is necessary and important to providing the essential information required for this critical comparison of "simulation" vs "experiment." In principle, the approach could be extended to higher spatial resolution and to include the effects of anesthetics on the electromechanical coupling mechanism in VGICs. © 2018 Elsevier Inc. All rights reserved.

Deforestation and Vectorial Capacity of Anopheles gambiae Giles Mosquitoes in Malaria Transmission, Kenya

PubMed Central

Afrane, Yaw A.; Little, Tom J.; Lawson, Bernard W.; Githeko, Andrew K.

2008-01-01

We investigated the effects of deforestation on microclimates and sporogonic development of Plasmodium falciparum parasites in Anopheles gambiae mosquitoes in an area of the western Kenyan highland prone to malaria epidemics. An. gambiae mosquitoes were fed with P. falciparum–infected blood through membrane feeders. Fed mosquitoes were placed in houses in forested and deforested areas in a highland area (1,500 m above sea level) and monitored for parasite development. Deforested sites had higher temperatures and relative humidities, and the overall infection rate of mosquitoes was increased compared with that in forested sites. Sporozoites appeared on average 1.1 days earlier in deforested areas. Vectorial capacity was estimated to be 77.7% higher in the deforested site than in the forested site. We showed that deforestation changes microclimates, leading to more rapid sporogonic development of P. falciparum and to a marked increase of malaria risk in the western Kenyan highland. PMID:18826815

Infection of Polarized Cultures of Human Intestinal Epithelial Cells with Hepatitis A Virus: Vectorial Release of Progeny Virions through Apical Cellular Membranes

PubMed Central

Blank, Christian A.; Anderson, David A.; Beard, Michael; Lemon, Stanley M.

2000-01-01

Although hepatitis A virus (HAV) is typically transmitted by the fecal-oral route, little is known of its interactions with cells of the gastrointestinal tract. We studied the replication of HAV in polarized cultures of Caco-2 cells, a human cell line which retains many differentiated functions of small intestinal epithelial cells. Virus uptake was 30- to 40-fold more efficient when the inoculum was placed on the apical rather than the basolateral surface of these cells, suggesting a greater abundance of the cellular receptor for HAV on the apical surface. Infection proceeded without cytopathic effect and did not influence transepithelial resistance or the diffusion of inulin across cell monolayers. Nonetheless, there was extensive release of progeny virus, which occurred almost exclusively into apical supernatant fluids (36.4% ± 12.5% of the total virus yield compared with 0.23% ± 0.13% release into basolateral fluids). Brefeldin A caused a profound inhibition of HAV replication, but also selectively reduced apical release of virus. These results indicate that polarized human epithelial cell cultures undergo vectorial infection with HAV and that virus release is largely restricted to the apical membrane. Virus release occurs in the absence of cytopathic effect and may involve cellular vesicular transport mechanisms. PMID:10864660

Ion pump sorting in polarized renal epithelial cells.

PubMed

Caplan, M J

2001-08-01

The plasma membranes of renal epithelial cells are divided into distinct apical and basolateral domains, which contain different inventories of ion transport proteins. Without this polarity vectorial ion and fluid transport would not be possible. Little is known of the signals and mechanisms that renal epithelial cells use to establish and maintain polarized distributions of their ion transport proteins. Analysis of ion pump sorting reveals that multiple complex signals participate in determining and regulating these proteins' subcellular localizations.

Self-assembled tethered bimolecular lipid membranes.

PubMed

Sinner, Eva-Kathrin; Ritz, Sandra; Naumann, Renate; Schiller, Stefan; Knoll, Wolfgang

2009-01-01

This chapter describes some of the strategies developed in our group for designing, constructing and structurally and functionally characterizing tethered bimolecular lipid membranes (tBLM). We introduce this platform as a novel model membrane system that complements the existing ones, for example, Langmuir monolayers, vesicular liposomal dispersions and bimolecular ("black") lipid membranes. Moreover, it offers the additional advantage of allowing for studies of the influence of membrane structure and order on the function of integral proteins, for example, on how the composition and organization of lipids in a mixed membrane influence the ion translocation activity of integral channel proteins. The first strategy that we introduce concerns the preparation of tethered monolayers by the self-assembly of telechelics. Their molecular architecture with a headgroup, a spacer unit (the "tether") and the amphiphile that mimics the lipid molecule allows them to bind specifically to the solid support thus forming the proximal layer of the final architecture. After fusion of vesicles that could contain reconstituted proteins from a liposomal dispersion in contact to this monolayer the tethered bimolecular lipid membrane is obtained. This can then be characterized by a broad range of surface analytical techniques, including surface plasmon spectroscopies, the quartz crystal microbalance, fluorescence and IR spectroscopies, and electrochemical techniques, to mention a few. It is shown that this concept allows for the construction of tethered lipid bilayers with outstanding electrical properties including resistivities in excess of 10 MOmega cm2. A modified strategy uses the assembly of peptides as spacers that couple covalently via their engineered sulfhydryl or lipoic acid groups at the N-terminus to the employed gold substrate, while their C-terminus is being activated afterward for the coupling of, for example, dimyristoylphosphatidylethanol amine (DMPE) lipid molecules via the NH2 moiety of their headgroups. It is demonstrated that these membranes are well suited for the in situ synthesis of membrane protein by a cell-free expression approach. The vectorial integration of an in vitro synthesized odorant receptor, OR5 from the rat, is demonstrated by means of antibodies that specifically bind to a tag at the N-terminus of the receptor and is read out by surface plasmon fluorescence spectroscopy. A completely different strategy employs his-tagged membrane proteins in their solubilized form binding to a surface-attached Ni(+)-NTA monolayer generating a well-oriented protein layer the density of which can be easily controlled by online monitoring the binding (assembly) step by surface plasmon spectroscopy. Moreover, the attachment of the his-tag to either the C- or the N-terminus allows for the complete control of the protein orientation. After the exchange of the detergent micelle by a lipid bilayer via a surface dialysis procedure an electrically very well isolating protein-tethered membrane is formed. We show that this "wiring" of the functional units allows for the (external) manipulation of the oxidation state of the redox-protein cytochrome c Oxidase by the control of the potential applied to the gold substrate which is used as the working electrode in an electrochemical attachment.

Oriented MOF-polymer Composite Nanofiber Membranes for High Proton Conductivity at High Temperature and Anhydrous Condition

PubMed Central

Wu, Bin; Pan, Jiefeng; Ge, Liang; Wu, Liang; Wang, Huanting; Xu, Tongwen

2014-01-01

The novel oriented electrospun nanofiber membrane composed of MOFs and SPPESK has been synthesized for proton exchange membrane fuel cell operating at high temperature and anhydrous conditions. It is clear that the oriented nanofiber membrane displays the higher proton conductivity than that of the disordered nanofiber membrane or the membrane prepared by conventional solvent-casting method (without nanofibers). Nanofibers within the membranes are significantly oriented. The proton conductivity of the oriented nanofiber membrane can reach up to (8.2 ± 0.16) × 10−2 S cm−1 at 160°C under anhydrous condition for the highly orientation of nanofibers. Moreover, the oxidative stability and resistance of methanol permeability of the nanofibers membrane are obviously improved with an increase in orientation of nanofibers. The observed methanol permeability of 0.707 × 10−7 cm2 s−1 is about 6% of Nafion-115. Consequently, orientated nanofibers membrane is proved to be a promising material as the proton exchange membrane for potential application in direct methanol fuel cells. PMID:25082522

Vectorial nanoscale mapping of optical antenna fields by single molecule dipoles.

PubMed

Singh, Anshuman; Calbris, Gaëtan; van Hulst, Niek F

2014-08-13

Optical nanoantennas confine light on the nanoscale, enabling strong light-matter interactions and ultracompact optical devices. Such confined nanovolumes of light have nonzero field components in all directions (x, y, and z). Unfortunately mapping of the actual nanoscale field vectors has so far remained elusive, though antenna hotspots have been explored by several techniques. In this paper, we present a novel method to probe all three components of the local antenna field. To this end a resonant nanoantenna is fabricated at the vertex of a scanning tip. Next, the nanoantenna is deterministically scanned in close proximity to single fluorescent molecules, whose fixed excitation dipole moment reads out the local field vector. With nanometer molecular resolution, we distinctly map x-, y-, and z-field components of the dipole antenna, i.e. a full vectorial mode map, and show good agreement with full 3D FDTD simulations. Moreover, the fluorescence polarization maps the localized coupling, with emission through the longitudinal antenna mode. Finally, the resonant antenna probe is used for single molecule imaging with 40 nm fwhm response function. The total fluorescence enhancement is 7.6 times, while out-of-plane molecules, almost undetectable in far-field, are made visible by the strong antenna z-field with a fluorescence enhancement up to 100 times. Interestingly, the apparent position of molecules shifts up to 20 nm depending on their orientation. The capability to resolve orientational information on the single molecule level makes the scanning resonant antenna an ideal tool for extreme resolution bioimaging.

Coherence and visibility for vectorial light.

PubMed

Luis, Alfredo

2010-08-01

Two-path interference of transversal vectorial waves is embedded within a larger scheme: this is four-path interference between four scalar waves. This comprises previous approaches to coherence between vectorial waves and restores the equivalence between correlation-based coherence and visibility.

Lubricant Jet Flow Phenomena in Spur and Helical Gears with Modified Addendums; for Radially Directed Individual Jets

NASA Technical Reports Server (NTRS)

Akin, Lee S.; Townsend, Dennis P.

1989-01-01

This paper develops the mathematical relations for the Virtual Kinetic Model as an improvement over the vectorial model developed earlier. The model solution described provides the most energy efficient means of cooling gears, i.e., it requires the least pressure or pumping power to distribute the coolant onto the tooth surface. Further, this nozzle orientation allows impingement to the root of the tooth if needed and provides the most cooling control when compared to into-mesh and out-of-mesh cooling.

Lubricant jet flow phenomena in spur and helical gears with modified addendums - For radially directed individual jets

NASA Technical Reports Server (NTRS)

Akin, L. S.; Townsend, D. P.

1989-01-01

This paper develops the mathematical relations for the Virtual Kinetic Model as an improvement over the vectorial model developed earlier. The model solution described provides the most energy efficient means of cooling gears, i.e., it requires the least pressure or pumping power to distribute the coolant onto the tooth surface. Further, this nozzle orientation allows impingement to the root of the tooth if needed and provides the most cooling control when compared to into-mesh and out-of-mesh cooling.

NMR studies reveal the role of biomembranes in modulating ligand binding and release by intracellular bile acid binding proteins.

PubMed

Pedò, Massimo; Löhr, Frank; D'Onofrio, Mariapina; Assfalg, Michael; Dötsch, Volker; Molinari, Henriette

2009-12-18

Bile acid molecules are transferred vectorially between basolateral and apical membranes of hepatocytes and enterocytes in the context of the enterohepatic circulation, a process regulating whole body lipid homeostasis. This work addresses the role of the cytosolic lipid binding proteins in the intracellular transfer of bile acids between different membrane compartments. We present nuclear magnetic resonance (NMR) data describing the ternary system composed of the bile acid binding protein, bile acids, and membrane mimetic systems, such as anionic liposomes. This work provides evidence that the investigated liver bile acid binding protein undergoes association with the anionic membrane and binding-induced partial unfolding. The addition of the physiological ligand to the protein-liposome mixture is capable of modulating this interaction, shifting the equilibrium towards the free folded holo protein. An ensemble of NMR titration experiments, based on nitrogen-15 protein and ligand observation, confirm that the membrane and the ligand establish competing binding equilibria, modulating the cytoplasmic permeability of bile acids. These results support a mechanism of ligand binding and release controlled by the onset of a bile salt concentration gradient within the polarized cell. The location of a specific protein region interacting with liposomes is highlighted.

Monitoring the function of membrane transport proteins in detergent-solubilized form

PubMed Central

Quick, Matthias; Javitch, Jonathan A.

2007-01-01

Transport proteins constitute ≈10% of most proteomes and play vital roles in the translocation of solutes across membranes of all organisms. Their (dys)function is implicated in many disorders, making them frequent targets for pharmacotherapy. The identification of substrates for members of this large protein family, still replete with many orphans of unknown function, has proven difficult, in part because high-throughput screening is greatly complicated by endogenous transporters present in many expression systems. In addition, direct structural studies require that transporters be extracted from the membrane with detergent, thereby precluding transport measurements because of the lack of a vectorial environment and necessitating reconstitution into proteoliposomes for activity measurements. Here, we describe a direct scintillation proximity-based radioligand-binding assay for determining transport protein function in crude cell extracts and in purified form. This rapid and universally applicable assay with advantages over cell-based platforms will greatly facilitate the identification of substrates for many orphan transporters and allows monitoring the function of transport proteins in a nonmembranous environment. PMID:17360689

Myristoylation Restricts Orientation of the GRASP Domain on Membranes and Promotes Membrane Tethering*

PubMed Central

Heinrich, Frank; Nanda, Hirsh; Goh, Haw Zan; Bachert, Collin; Lösche, Mathias; Linstedt, Adam D.

2014-01-01

The mammalian Golgi reassembly stacking protein (GRASP) proteins are Golgi-localized homotypic membrane tethers that organize Golgi stacks into a long, contiguous ribbon-like structure. It is unknown how GRASPs undergo trans pairing given that cis interactions between the proteins in the plane of the membrane are intrinsically favored. To test the hypothesis that myristoylation of the self-interacting GRASP domain restricts its orientation on the membrane to favor trans pairing, we established an in vitro assay that recapitulates GRASP-dependent membrane tethering and used neutron reflection under similar conditions to determine the orientation of the GRASP domain. In vivo, the membrane association of GRASP proteins is conferred by the simultaneous insertion of an N-terminal myristic acid and binding to a Golgi-associated binding partner. In our assay, the latter contact was replaced using a C-terminal hexa-His moiety, which bound to Ni2+-conjugated lipids incorporated into a substrate-supported bilayer lipid membrane. Nonmyristoylated protein lacked a fixed orientation on the membrane and inefficiently tethered liposomes. In contrast, myristoylated GRASP promoted tethering and exhibited a unique membrane complex. Thus, myristoylation restricts the membrane orientation of the GRASP domain favoring interactions in trans for membrane tethering. PMID:24505136

High precision wavefront control in point spread function engineering for single emitter localization

NASA Astrophysics Data System (ADS)

Siemons, M.; Hulleman, C. N.; Thorsen, R. Ø.; Smith, C. S.; Stallinga, S.

2018-04-01

Point Spread Function (PSF) engineering is used in single emitter localization to measure the emitter position in 3D and possibly other parameters such as the emission color or dipole orientation as well. Advanced PSF models such as spline fits to experimental PSFs or the vectorial PSF model can be used in the corresponding localization algorithms in order to model the intricate spot shape and deformations correctly. The complexity of the optical architecture and fit model makes PSF engineering approaches particularly sensitive to optical aberrations. Here, we present a calibration and alignment protocol for fluorescence microscopes equipped with a spatial light modulator (SLM) with the goal of establishing a wavefront error well below the diffraction limit for optimum application of complex engineered PSFs. We achieve high-precision wavefront control, to a level below 20 m$\\lambda$ wavefront aberration over a 30 minute time window after the calibration procedure, using a separate light path for calibrating the pixel-to-pixel variations of the SLM, and alignment of the SLM with respect to the optical axis and Fourier plane within 3 $\\mu$m ($x/y$) and 100 $\\mu$m ($z$) error. Aberrations are retrieved from a fit of the vectorial PSF model to a bead $z$-stack and compensated with a residual wavefront error comparable to the error of the SLM calibration step. This well-calibrated and corrected setup makes it possible to create complex `3D+$\\lambda$' PSFs that fit very well to the vectorial PSF model. Proof-of-principle bead experiments show precisions below 10~nm in $x$, $y$, and $\\lambda$, and below 20~nm in $z$ over an axial range of 1 $\\mu$m with 2000 signal photons and 12 background photons.

Electrostatic interactions and binding orientation of HIV-1 matrix studied by neutron reflectivity.

PubMed

Nanda, Hirsh; Datta, Siddhartha A K; Heinrich, Frank; Lösche, Mathias; Rein, Alan; Krueger, Susan; Curtis, Joseph E

2010-10-20

The N-terminal matrix (MA) domain of the HIV-1 Gag protein is responsible for binding to the plasma membrane of host cells during viral assembly. The putative membrane-binding interface of MA was previously mapped by means of mutagenesis and analysis of its trimeric crystal structure. However, the orientation of MA on membranes has not been directly determined by experimental measurements. We present neutron reflectivity measurements that resolve the one-dimensional scattering length density profile of MA bound to a biomimetic of the native viral membrane. A molecular refinement procedure was developed using atomic structures of MA to determine the orientation of the protein on the membrane. The orientation defines a lipid-binding interface consistent with previous mutagenesis results. The MA protein maintains this orientation without the presence of a myristate group, driven only by electrostatic interactions. Furthermore, MA is found to penetrate the membrane headgroup region peripherally such that only the side chains of specific Lys and Arg residues interact with the surface. The results suggest that electrostatic interactions are sufficient to favorably orient MA on viral membrane mimics. The spatial determination of the membrane-bound protein demonstrates the ability of neutron reflectivity to discern orientation and penetration under physiologically relevant conditions. Copyright © 2010 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Beam shaping with vectorial vortex beams under low numerical aperture illumination condition

NASA Astrophysics Data System (ADS)

Dai, Jianning; Zhan, Qiwen

2008-08-01

In this paper we propose and demonstrate a novel beam shaping method using vectorial vortex beam. A vectorial vortex beam is laser beam with polarization singularity in the beam cross section. This type of beams can be decomposed into two orthogonally polarized components. Each of the polarized components could have different vortex characteristics, and consequently, different intensity distribution when focused by lens. Beam shaping in the far field can be achieved by adjusting the relative weighing of these two components. As one example, we study the vectorial vortex that consists of a linearly polarized Gaussian component and a vortex component polarized orthogonally. When such a vectorial vortex beam is focus by low NA lens, the Gaussian component gives rise to a focal intensity distribution with a solid centre while the vortex component gives rise to a donut distribution with hollow dark center. The shape of the focus can be continuously varied by continuously adjusting the relative weight of the two components. Under appropriate conditions, flat top focusing can be obtained. We experimentally demonstrate the creation of such beams with a liquid crystal spatial light modulator. Flattop focus obtained by vectorial vortex beams with topological charge of +1 has been obtained.

Vectorial Entry and Release of Hepatitis A Virus in Polarized Human Hepatocytes ▿

PubMed Central

Snooks, Michelle J.; Bhat, Purnima; Mackenzie, Jason; Counihan, Natalie A.; Vaughan, Nicola; Anderson, David A.

2008-01-01

Hepatitis A virus (HAV) is an enterically transmitted virus that replicates predominantly in hepatocytes within the liver before excretion via bile through feces. Hepatocytes are polarized epithelial cells, and it has been assumed that the virus load in bile results from direct export of HAV via the apical domain of polarized hepatocytes. We have developed a subclone of hepatocyte-derived HepG2 cells (clone N6) that maintains functional characteristics of polarized hepatocytes but displays morphology typical of columnar epithelial cells, rather than the complex morphology that is typical of hepatocytes. N6 cells form microcolonies of polarized cells when grown on glass and confluent monolayers of polarized cells on semipermeable membranes. When N6 microcolonies were exposed to HAV, infection was restricted to peripheral cells of polarized colonies, whereas all cells could be infected in colonies of nonpolarized HepG2 cells (clone C11) or following disruption of tight junctions in N6 colonies with EGTA. This suggests that viral entry occurs predominantly via the basolateral plasma membrane, consistent with uptake of virus from the bloodstream after enteric exposure, as expected. Viral export was also found to be markedly vectorial in N6 but not C11 cells. However, rather than being exported from the apical domain as expected, more than 95% of HAV was exported via the basolateral domain of N6 cells, suggesting that virus is first excreted from infected hepatocytes into the bloodstream rather than to the biliary tree. Enteric excretion of HAV may therefore rely on reuptake and transcytosis of progeny HAV across hepatocytes into the bile. These studies provide the first example of the interactions between viruses and polarized hepatocytes. PMID:18579610

Vectorial strain gauge method using single flexible orthogonal polydimethylsiloxane gratings

NASA Astrophysics Data System (ADS)

Guo, Hao; Tang, Jun; Qian, Kun; Tsoukalas, Dimitris; Zhao, Miaomiao; Yang, Jiangtao; Zhang, Binzhen; Chou, Xiujian; Liu, Jun; Xue, Chenyang; Zhang, Wendong

2016-03-01

A vectorial strain gauge method using a single sensing element is reported based on the double-sided polydimethylsiloxane (PDMS) Fraunhofer diffraction gratings structures. Using O2 plasma treatment steps, orthogonal wrinkled gratings were fabricated on both sides of a pre-strained PDMS film. Diffracted laser spots from this structure have been used to experimentally demonstrate, that any applied strain can be quantitatively characterized in both the x and y directions with an error of less than 0.6% and with a gauge factor of approximately 10. This simple and low cost technology which is completely different from the traditional vectorial strain gauge method, can be applied to surface vectorial strain measurement and multi-axis integrated mechanical sensors.

Determining the Orientation and Localization of Membrane-Bound Peptides

PubMed Central

Hohlweg, Walter; Kosol, Simone; Zangger, Klaus

2012-01-01

Many naturally occurring bioactive peptides bind to biological membranes. Studying and elucidating the mode of interaction is often an essential step to understand their molecular and biological functions. To obtain the complete orientation and immersion depth of such compounds in the membrane or a membrane-mimetic system, a number of methods are available, which are separated in this review into four main classes: solution NMR, solid-state NMR, EPR and other methods. Solution NMR methods include the Nuclear Overhauser Effect (NOE) between peptide and membrane signals, residual dipolar couplings and the use of paramagnetic probes, either within the membrane-mimetic or in the solvent. The vast array of solid state NMR methods to study membrane-bound peptide orientation and localization includes the anisotropic chemical shift, PISA wheels, dipolar waves, the GALA, MAOS and REDOR methods and again the use of paramagnetic additives on relaxation rates. Paramagnetic additives, with their effect on spectral linewidths, have also been used in EPR spectroscopy. Additionally, the orientation of a peptide within a membrane can be obtained by the anisotropic hyperfine tensor of a rigidly attached nitroxide label. Besides these magnetic resonance techniques a series of other methods to probe the orientation of peptides in membranes has been developed, consisting of fluorescence-, infrared- and oriented circular dichroism spectroscopy, colorimetry, interface-sensitive X-ray and neutron scattering and Quartz crystal microbalance. PMID:22044140

Fluorescence Imaging of Two-Photon Linear Dichroism: Cholesterol Depletion Disrupts Molecular Orientation in Cell Membranes

PubMed Central

Benninger, Richard K. P.; Önfelt, Björn; Neil, Mark A. A.; Davis, Daniel M.; French, Paul M. W.

2005-01-01

The plasma membrane of cells is an ordered environment, giving rise to anisotropic orientation and restricted motion of molecules and proteins residing in the membrane. At the same time as being an organized matrix of defined structure, the cell membrane is heterogeneous and dynamic. Here we present a method where we use fluorescence imaging of linear dichroism to measure the orientation of molecules relative to the cell membrane. By detecting linear dichroism as well as fluorescence anisotropy, the orientation parameters are separated from dynamic properties such as rotational diffusion and homo energy transfer (energy migration). The sensitivity of the technique is enhanced by using two-photon excitation for higher photo-selection compared to single photon excitation. We show here that we can accurately image lipid organization in whole cell membranes and in delicate structures such as membrane nanotubes connecting two cells. The speed of our wide-field imaging system makes it possible to image changes in orientation and anisotropy occurring on a subsecond timescale. This is demonstrated by time-lapse studies showing that cholesterol depletion rapidly disrupts the orientation of a fluorophore located within the hydrophobic region of the cell membrane but not of a surface bound probe. This is consistent with cholesterol having an important role in stabilizing and ordering the lipid tails within the plasma membrane. PMID:15520272

Laser printed nano-gratings: orientation and period peculiarities

NASA Astrophysics Data System (ADS)

Stankevič, Valdemar; Račiukaitis, Gediminas; Bragheri, Francesca; Wang, Xuewen; Gamaly, Eugene G.; Osellame, Roberto; Juodkazis, Saulius

2017-01-01

Understanding of material behaviour at nanoscale under intense laser excitation is becoming critical for future application of nanotechnologies. Nanograting formation by linearly polarised ultra-short laser pulses has been studied systematically in fused silica for various pulse energies at 3D laser printing/writing conditions, typically used for the industrial fabrication of optical elements. The period of the nanogratings revealed a dependence on the orientation of the scanning direction. A tilt of the nanograting wave vector at a fixed laser polarisation was also observed. The mechanism responsible for this peculiar dependency of several features of the nanogratings on the writing direction is qualitatively explained by considering the heat transport flux in the presence of a linearly polarised electric field, rather than by temporal and spatial chirp of the laser beam. The confirmed vectorial nature of the light-matter interaction opens new control of material processing with nanoscale precision.

Seipin performs dissectible functions in promoting lipid droplet biogenesis and regulating droplet morphology

PubMed Central

Cartwright, Bethany R.; Binns, Derk D.; Hilton, Christopher L.; Han, Sungwon; Gao, Qiang; Goodman, Joel M.

2015-01-01

Seipin is necessary for both adipogenesis and lipid droplet (LD) organization in nonadipose tissues; however, its molecular function is incompletely understood. Phenotypes in the seipin-null mutant of Saccharomyces cerevisiae include aberrant droplet morphology (endoplasmic reticulum–droplet clusters and size heterogeneity) and sensitivity of droplet size to changes in phospholipid synthesis. It has not been clear, however, whether seipin acts in initiation of droplet synthesis or at a later step. Here we utilize a system of de novo droplet formation to show that the absence of seipin results in a delay in droplet appearance with concomitant accumulation of neutral lipid in membranes. We also demonstrate that seipin is required for vectorial budding of droplets toward the cytoplasm. Furthermore, we find that the normal rate of droplet initiation depends on 14 amino acids at the amino terminus of seipin, deletion of which results in fewer, larger droplets that are consistent with a delay in initiation but are otherwise normal in morphology. Importantly, other functions of seipin, namely vectorial budding and resistance to inositol, are retained in this mutant. We conclude that seipin has dissectible roles in both promoting early LD initiation and in regulating LD morphology, supporting its importance in LD biogenesis. PMID:25540432

CW dipolar broadening EPR spectroscopy and mechanically aligned bilayers used to measure distance and relative orientation between two TOAC spin labels on an antimicrobial peptide

NASA Astrophysics Data System (ADS)

Sahu, Indra D.; Hustedt, Eric J.; Ghimire, Harishchandra; Inbaraj, Johnson J.; McCarrick, Robert M.; Lorigan, Gary A.

2014-12-01

An EPR membrane alignment technique was applied to measure distance and relative orientations between two spin labels on a protein oriented along the surface of the membrane. Previously we demonstrated an EPR membrane alignment technique for measuring distances and relative orientations between two spin labels using a dual TOAC-labeled integral transmembrane peptide (M2δ segment of Acetylcholine receptor) as a test system. In this study we further utilized this technique and successfully measured the distance and relative orientations between two spin labels on a membrane peripheral peptide (antimicrobial peptide magainin-2). The TOAC-labeled magainin-2 peptides were mechanically aligned using DMPC lipids on a planar quartz support, and CW-EPR spectra were recorded at specific orientations. Global analysis in combination with rigorous spectral simulation was used to simultaneously analyze data from two different sample orientations for both single- and double-labeled peptides. We measured an internitroxide distance of 15.3 Å from a dual TOAC-labeled magainin-2 peptide at positions 8 and 14 that closely matches with the 13.3 Å distance obtained from a model of the labeled magainin peptide. In addition, the angles determining the relative orientations of the two nitroxides have been determined, and the results compare favorably with molecular modeling. This study demonstrates the utility of the technique for proteins oriented along the surface of the membrane in addition to the previous results for proteins situated within the membrane bilayer.

Extracellular pH regulation in microdomains of colonic crypts: effects of short-chain fatty acids.

PubMed Central

Chu, S; Montrose, M H

1995-01-01

It has been suggested that transepithelial gradients of short-chain fatty acids (SCFAs; the major anions in the colonic lumen) generate pH gradients across the colonic epithelium. Quantitative confocal microscopy was used to study extracellular pH in mouse distal colon with intact epithelial architecture, by superfusing tissue with carboxy SNARF-1 (a pH-sensitive fluorescent dye). Results demonstrate extracellular pH regulation in two separate microdomains surrounding colonic crypts: the crypt lumen and the subepithelial tissue adjacent to crypt colonocytes. Apical superfusion with (i) a poorly metabolized SCFA (isobutyrate), (ii) an avidly metabolized SCFA (n-butyrate), or (iii) a physiologic mixture of acetate/propionate/n-butyrate produced similar results: alkalinization of the crypt lumen and acidification of subepithelial tissue. Effects were (i) dependent on the presence and orientation of a transepithelial SCFA gradient, (ii) not observed with gluconate substitution, and (iii) required activation of sustained vectorial acid/base transport by SCFAs. Results suggest that the crypt lumen functions as a pH microdomain due to slow mixing with bulk superfusates and that crypts contribute significant buffering capacity to the lumen. In conclusion, physiologic SCFA gradients cause polarized extracellular pH regulation because epithelial architecture and vectorial transport synergize to establish regulated microenvironments. Images Fig. 1 Fig. 3 PMID:7724557

Oriented molecular sieve membranes by heteroepitaxial growth.

PubMed

Jeong, Hae-Kwon; Krohn, John; Sujaoti, Khristina; Tsapatsis, Michael

2002-11-06

Heteroepitaxial growth of titanosilicates (ETS-10 and ETS-4) is reported. Using this heteroepitaxial growth, oriented ETS-10/-4 membranes have been fabricated, demonstrating a novel way to achieve preferred orientation of molecular sieve films.

Chromophore orientation in bacteriorhodopsin determined from the angular dependence of deuterium nuclear magnetic resonance spectra of oriented purple membranes.

PubMed

Moltke, S; Nevzorov, A A; Sakai, N; Wallat, I; Job, C; Nakanishi, K; Heyn, M P; Brown, M F

1998-08-25

The orientation of prosthetic groups in membrane proteins is of considerable importance in understanding their functional role in energy conversion, signal transduction, and ion transport. In this work, the orientation of the retinylidene chromophore of bacteriorhodopsin (bR) was investigated using 2H NMR spectroscopy. Bacteriorhodopsin was regenerated with all-trans-retinal stereospecifically deuterated in one of the geminal methyl groups on C1 of the cyclohexene ring. A highly oriented sample, which is needed to obtain individual bond orientations from 2H NMR, was prepared by forming hydrated lamellar films of purple membranes on glass slides. A Monte Carlo method was developed to accurately simulate the 2H NMR line shape due to the distribution of bond angles and the orientational disorder of the membranes. The number of free parameters in the line shape simulation was reduced by independent measurements of the intrinsic line width (1.6 kHz from T2e experiments) and the effective quadrupolar coupling constant (38. 8-39.8 kHz from analysis of the line shape of a powder-type sample). The angle between the C1-(1R)-1-CD3 bond and the purple membrane normal was determined with high accuracy from the simultaneous analysis of a series of 2H NMR spectra recorded at different inclinations of the uniaxially oriented sample in the magnetic field at 20 and -50 degrees C. The value of 68.7 +/- 2.0 degrees in dark-adapted bR was used, together with the previously determined angle of the C5-CD3 bond, to calculate the possible orientations of the cyclohexene ring in the membrane. The solutions obtained from 2H NMR were then combined with additional constraints from linear dichroism and electron cryomicroscopy to obtain the allowed orientations of retinal in the noncentrosymmetric membrane structure. The combined data indicate that the methyl groups on the polyene chain point toward the cytoplasmic side of the membrane and the N-H bond of the Schiff base to the extracellular side, i.e., toward the side of proton release in the pump pathway.

Effect of Larval Competition on Extrinsic Incubation Period and Vectorial Capacity of Aedes albopictus for Dengue Virus.

PubMed

Bara, Jeffrey; Rapti, Zoi; Cáceres, Carla E; Muturi, Ephantus J

2015-01-01

Despite the growing awareness that larval competition can influence adult mosquito life history traits including susceptibility to pathogens, the net effect of larval competition on human risk of exposure to mosquito-borne pathogens remains poorly understood. We examined how intraspecific larval competition affects dengue-2 virus (DENV-2) extrinsic incubation period and vectorial capacity of its natural vector Aedes albopictus. Adult Ae. albopictus from low and high-larval density conditions were orally challenged with DENV-2 and then assayed for virus infection and dissemination rates following a 6, 9, or 12-day incubation period using real-time quantitative reverse transcription PCR. We then modeled the effect of larval competition on vectorial capacity using parameter estimates obtained from peer-reviewed field and laboratory studies. Larval competition resulted in significantly longer development times, lower emergence rates, and smaller adults, but did not significantly affect the extrinsic incubation period of DENV-2 in Ae. albopictus. Our vectorial capacity models suggest that the effect of larval competition on adult mosquito longevity likely has a greater influence on vectorial capacity relative to any competition-induced changes in vector competence. Furthermore, we found that large increases in the viral dissemination rate may be necessary to compensate for small competition-induced reductions in daily survivorship. Our results indicate that mosquito populations that experience stress from larval competition are likely to have a reduced vectorial capacity, even when susceptibility to pathogens is enhanced.

Effect of Larval Competition on Extrinsic Incubation Period and Vectorial Capacity of Aedes albopictus for Dengue Virus

PubMed Central

Bara, Jeffrey; Rapti, Zoi; Cáceres, Carla E.; Muturi, Ephantus J.

2015-01-01

Despite the growing awareness that larval competition can influence adult mosquito life history traits including susceptibility to pathogens, the net effect of larval competition on human risk of exposure to mosquito-borne pathogens remains poorly understood. We examined how intraspecific larval competition affects dengue-2 virus (DENV-2) extrinsic incubation period and vectorial capacity of its natural vector Aedes albopictus. Adult Ae. albopictus from low and high-larval density conditions were orally challenged with DENV-2 and then assayed for virus infection and dissemination rates following a 6, 9, or 12-day incubation period using real-time quantitative reverse transcription PCR. We then modeled the effect of larval competition on vectorial capacity using parameter estimates obtained from peer-reviewed field and laboratory studies. Larval competition resulted in significantly longer development times, lower emergence rates, and smaller adults, but did not significantly affect the extrinsic incubation period of DENV-2 in Ae. albopictus. Our vectorial capacity models suggest that the effect of larval competition on adult mosquito longevity likely has a greater influence on vectorial capacity relative to any competition-induced changes in vector competence. Furthermore, we found that large increases in the viral dissemination rate may be necessary to compensate for small competition-induced reductions in daily survivorship. Our results indicate that mosquito populations that experience stress from larval competition are likely to have a reduced vectorial capacity, even when susceptibility to pathogens is enhanced. PMID:25951173

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

PubMed

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

2018-03-06

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

Cooperative folding of a polytopic α-helical membrane protein involves a compact N-terminal nucleus and nonnative loops

PubMed Central

Paslawski, Wojciech; Lillelund, Ove K.; Kristensen, Julie Veje; Schafer, Nicholas P.; Baker, Rosanna P.; Urban, Sinisa; Otzen, Daniel E.

2015-01-01

Despite the ubiquity of helical membrane proteins in nature and their pharmacological importance, the mechanisms guiding their folding remain unclear. We performed kinetic folding and unfolding experiments on 69 mutants (engineered every 2–3 residues throughout the 178-residue transmembrane domain) of GlpG, a membrane-embedded rhomboid protease from Escherichia coli. The only clustering of significantly positive ϕ-values occurs at the cytosolic termini of transmembrane helices 1 and 2, which we identify as a compact nucleus. The three loops flanking these helices show a preponderance of negative ϕ-values, which are sometimes taken to be indicative of nonnative interactions in the transition state. Mutations in transmembrane helices 3–6 yielded predominantly ϕ-values near zero, indicating that this part of the protein has denatured-state–level structure in the transition state. We propose that loops 1–3 undergo conformational rearrangements to position the folding nucleus correctly, which then drives folding of the rest of the domain. A compact N-terminal nucleus is consistent with the vectorial nature of cotranslational membrane insertion found in vivo. The origin of the interactions in the transition state that lead to a large number of negative ϕ-values remains to be elucidated. PMID:26056273

Equilibrium of fluid membranes endowed with orientational order

NASA Astrophysics Data System (ADS)

Kumar Alageshan, Jaya; Chakrabarti, Buddhapriya; Hatwalne, Yashodhan

2017-04-01

Minimization of the low-temperature elastic free-energy functional of orientationlly ordered membranes involves independent variation of the membrane-shape, while keeping the orientational order on it (its texture) fixed. We propose an operational, coordinate-independent method for implementing such a variation. Using the Nelson-Peliti formulation of elasticity that emphasizes the interplay between geometry, topology, and thermal fluctuations of orientationally ordered membranes, we minimize the elastic free energy to obtain equations governing their equilibrium shape, together with associated free boundary conditions. Our results are essential for understanding and predicting equilibrium shapes as well as textures of membranes and vesicles; particularly under conditions in which shape deformations are large.

Quantitative and Mechanistic Assessment of Model Lipophilic Drugs in Micellar Solutions in the Transport Kinetics Across MDR1-MDCK Cell Monolayers.

PubMed

Ho, Norman F H; Nielsen, James; Peterson, Michelle; Burton, Philip S

2016-02-01

An approach to characterizing P-glycoprotein (Pgp) interaction potential for sparingly water-soluble compounds was developed using bidirectional transport kinetics in MDR1-MDCK cell monolayers. Paclitaxel, solubilized in a dilute polysorbate 80 (PS80) micellar solution, was used as a practical example. Although the passage of paclitaxel across the cell monolayer was initially governed by the thermodynamic activity of the micelle-solubilized drug solution, Pgp inhibition was sustained by the thermodynamic activity (i.e., critical micelle concentration) of the PS80 micellar solution bathing the apical (ap) membrane. The mechanistic understanding of the experimental strategies and treatment of data was supported by a biophysical model expressed in the form of transport events occurring at the ap and basolateral (bl) membranes in series whereas the vectorial directions of the transcellular kinetics were accommodated. The derived equations permitted the stepwise quantitative delineation of the Pgp efflux activity (inhibited and uninhibited by PS80) and the passive permeability coefficient of the ap membrane, the passive permeability at the bl membrane and, finally, the distinct coupling of these with efflux pump activity to identify the rate-determining steps and mechanisms. The Jmax/KM(∗) for paclitaxel was in the order of 10(-4) cm/s and the ap- and bl-membrane passive permeability coefficients were asymmetric, with bl-membrane permeability significantly greater than ap. Copyright © 2016 American Pharmacists Association®. Published by Elsevier Inc. All rights reserved.

Determination of the polarization states of an arbitrary polarized terahertz beam: Vectorial vortex analysis

PubMed Central

Wakayama, Toshitaka; Higashiguchi, Takeshi; Oikawa, Hiroki; Sakaue, Kazuyuki; Washio, Masakazu; Yonemura, Motoki; Yoshizawa, Toru; Tyo, J. Scott; Otani, Yukitoshi

2015-01-01

Vectorial vortex analysis is used to determine the polarization states of an arbitrarily polarized terahertz (0.1–1.6 THz) beam using THz achromatic axially symmetric wave (TAS) plates, which have a phase retardance of Δ = 163° and are made of polytetrafluorethylene. Polarized THz beams are converted into THz vectorial vortex beams with no spatial or wavelength dispersion, and the unknown polarization states of the incident THz beams are reconstructed. The polarization determination is also demonstrated at frequencies of 0.16 and 0.36 THz. The results obtained by solving the inverse source problem agree with the values used in the experiments. This vectorial vortex analysis enables a determination of the polarization states of the incident THz beam from the THz image. The polarization states of the beams are estimated after they pass through the TAS plates. The results validate this new approach to polarization detection for intense THz sources. It could find application in such cutting edge areas of physics as nonlinear THz photonics and plasmon excitation, because TAS plates not only instantaneously elucidate the polarization of an enclosed THz beam but can also passively control THz vectorial vortex beams. PMID:25799965

Determination of the polarization states of an arbitrary polarized terahertz beam: vectorial vortex analysis.

PubMed

Wakayama, Toshitaka; Higashiguchi, Takeshi; Oikawa, Hiroki; Sakaue, Kazuyuki; Washio, Masakazu; Yonemura, Motoki; Yoshizawa, Toru; Tyo, J Scott; Otani, Yukitoshi

2015-03-24

Vectorial vortex analysis is used to determine the polarization states of an arbitrarily polarized terahertz (0.1-1.6 THz) beam using THz achromatic axially symmetric wave (TAS) plates, which have a phase retardance of Δ = 163° and are made of polytetrafluorethylene. Polarized THz beams are converted into THz vectorial vortex beams with no spatial or wavelength dispersion, and the unknown polarization states of the incident THz beams are reconstructed. The polarization determination is also demonstrated at frequencies of 0.16 and 0.36 THz. The results obtained by solving the inverse source problem agree with the values used in the experiments. This vectorial vortex analysis enables a determination of the polarization states of the incident THz beam from the THz image. The polarization states of the beams are estimated after they pass through the TAS plates. The results validate this new approach to polarization detection for intense THz sources. It could find application in such cutting edge areas of physics as nonlinear THz photonics and plasmon excitation, because TAS plates not only instantaneously elucidate the polarization of an enclosed THz beam but can also passively control THz vectorial vortex beams.

L'espace articulaire de la Robotique Industrielle est un espace vectorielIndustrial Robotics joint space is a vector space

NASA Astrophysics Data System (ADS)

Tondu, Bertrand

2003-05-01

The mathematical modelling of industrial robots is based on the vectorial nature of the n-dimensional joint space of the robot, defined as a kinematic chain with n degrees of freedom. However, in our opinion, the vectorial nature of the joint space has been insufficiently discussed in the literature. We establish the vectorial nature of the joint space of an industrial robot from the fundamental studies of B. Roth on screws. To cite this article: B. Tondu, C. R. Mecanique 331 (2003).

Drug Distribution. Part 1. Models to Predict Membrane Partitioning.

PubMed

Nagar, Swati; Korzekwa, Ken

2017-03-01

Tissue partitioning is an important component of drug distribution and half-life. Protein binding and lipid partitioning together determine drug distribution. Two structure-based models to predict partitioning into microsomal membranes are presented. An orientation-based model was developed using a membrane template and atom-based relative free energy functions to select drug conformations and orientations for neutral and basic drugs. The resulting model predicts the correct membrane positions for nine compounds tested, and predicts the membrane partitioning for n = 67 drugs with an average fold-error of 2.4. Next, a more facile descriptor-based model was developed for acids, neutrals and bases. This model considers the partitioning of neutral and ionized species at equilibrium, and can predict membrane partitioning with an average fold-error of 2.0 (n = 92 drugs). Together these models suggest that drug orientation is important for membrane partitioning and that membrane partitioning can be well predicted from physicochemical properties.

Magnetic orientation in C. elegans relies on the integrity of the villi of the AFD magnetosensory neurons.

PubMed

Bainbridge, Chance; Rodriguez, Anjelica; Schuler, Andrew; Cisneros, Michael; Vidal-Gadea, Andrés G

2016-10-01

The magnetic field of the earth provides many organisms with sufficient information to successfully navigate through their environments. While evidence suggests the widespread use of this sensory modality across many taxa, it remains an understudied sensory modality. We have recently showed that the nematode C. elegans orients to earth-strength magnetic fields using the first pair of described magnetosensory neurons, AFDs. The AFD cells are a pair of ciliated sensory neurons crowned by fifty villi known to be implicated in temperature sensation. We investigated the potential importance of these subcellular structures for the performance of magnetic orientation. We show that ciliary integrity and villi number are essential for magnetic orientation. Mutants with impairments AFD cilia or villi structure failed to orient to magnetic fields. Similarly, C. elegans larvae possessing immature AFD neurons with fewer villi were also unable to orient to magnetic fields. Larvae of every stage however retained the ability to orient to thermal gradients. To our knowledge, this is the first behavioral separation of magnetic and thermal orientation in C. elegans. We conclude that magnetic orientation relies on the function of both cilia and villi in the AFD neurons. The role of villi in multiple sensory transduction pathways involved in the sensory transduction of vectorial stimuli further supports the likely role of the villi of the AFD neurons as the site for magnetic field transduction. The genetic and behavioral tractability of C. elegans make it a promising system for uncovering potentially conserved molecular mechanisms by which animals across taxa detect and orient to magnetic fields. Copyright © 2016 Elsevier Ltd. All rights reserved.

Drug Transporters and Na+/H+ Exchange Regulatory Factor PSD-95/Drosophila Discs Large/ZO-1 Proteins

PubMed Central

Walsh, Dustin R.; Nolin, Thomas D.

2015-01-01

Drug transporters govern the absorption, distribution, and elimination of pharmacologically active compounds. Members of the solute carrier and ATP binding-cassette drug transporter family mediate cellular drug uptake and efflux processes, thereby coordinating the vectorial movement of drugs across epithelial barriers. To exert their physiologic and pharmacological function in polarized epithelia, drug transporters must be targeted and stabilized to appropriate regions of the cell membrane (i.e., apical versus basolateral). Despite the critical importance of drug transporter membrane targeting, the mechanisms that underlie these processes are largely unknown. Several clinically significant drug transporters possess a recognition sequence that binds to PSD-95/Drosophila discs large/ZO-1 (PDZ) proteins. PDZ proteins, such as the Na+/H+ exchanger regulatory factor (NHERF) family, act to stabilize and organize membrane targeting of multiple transmembrane proteins, including many clinically relevant drug transporters. These PDZ proteins are normally abundant at apical membranes, where they tether membrane-delimited transporters. NHERF expression is particularly high at the apical membrane in polarized tissue such as intestinal, hepatic, and renal epithelia, tissues important to drug disposition. Several recent studies have highlighted NHERF proteins as determinants of drug transporter function secondary to their role in controlling membrane abundance and localization. Mounting evidence strongly suggests that NHERF proteins may have clinically significant roles in pharmacokinetics and pharmacodynamics of several pharmacologically active compounds and may affect drug action in cancer and chronic kidney disease. For these reasons, NHERF proteins represent a novel class of post-translational mediators of drug transport and novel targets for new drug development. PMID:26092975

GO-guided direct growth of highly oriented metal-organic framework nanosheet membranes for H2/CO2 separation.

PubMed

Li, Yujia; Liu, Haiou; Wang, Huanting; Qiu, Jieshan; Zhang, Xiongfu

2018-05-07

Highly oriented, ultrathin metal-organic framework (MOF) membranes are attractive for practical separation applications, but the scalable preparation of such membranes especially on standard tubular supports remains a huge challenge. Here we report a novel bottom-up strategy for directly growing a highly oriented Zn 2 (bIm) 4 (bIm = benzimidazole) ZIF nanosheet tubular membrane, based on graphene oxide (GO) guided self-conversion of ZnO nanoparticles (NPs). Through our approach, a thin layer of ZnO NPs confined between a substrate and a GO ultrathin layer self-converts into a highly oriented Zn 2 (bIm) 4 nanosheet membrane. The resulting membrane with a thickness of around 200 nm demonstrates excellent H 2 /CO 2 gas separation performance with a H 2 performance of 1.4 × 10 -7 mol m -2 s -1 Pa -1 and an ideal separation selectivity of about 106. The method can be easily scaled up and extended to the synthesis of other types of Zn-based MOF nanosheet membranes. Importantly, our strategy is particularly suitable for the large-scale fabrication of tubular MOF membranes that has not been possible through other methods.

Effect of temperature on life history parameters of adult Culicoides sonorensis (Diptera: Ceratopogonidae) in relation to geographic origin and vectorial capacity for bluetongue virus.

PubMed

Lysyk, T J; Danyk, T

2007-09-01

The effect of temperature on survival, oviposition, gonotrophic development, and a life history factor of vectorial capacity were examined in adult Culicoides sonorensis (Wirth & Jones) (Diptera: Ceratopogonidae) that originated from two geographic locations. Flies originating from the United States (Colorado) had slightly reduced survival after a bloodmeal compared with wild flies collected in southern Alberta (AB), Canada. Survival of AB flies declined in a curvilinear manner with temperature, whereas survival of U.S. flies showed a linear response to temperature. The survivorship curve of the AB flies more closely followed a Weibull distribution than an exponential, indicating survival was age-dependent. Survivorship of the U.S. flies followed an exponential distribution. Females from both sources laid similar numbers of eggs throughout their life. The first eggs were laid by females from both sources at 31.9 degree-day (DD)9.3. Dissections of blood-fed flies reared at various temperatures indicated that flies from both sources were 90% gravid at 32 DD9.3. Relationships among temperature and life history components of vectorial capacity were similar among flies from the two sources and indicated that vectorial capacity would be approximately 1.8-2.6-fold greater in a southern U.S. climate compared with southwestern Canada due solely to the effects of temperature on the life history of C. sonorensis. Using life history estimates derived from Weibull model had little effect on estimating vectorial capacity, whereas using estimates derived from the exponential model slightly overestimated vectorial capacity.

Vectorial approach of determining the wave propagation at metasurfaces

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

Smith, Daniel, E-mail: D.Smith1966@outlook.com; Campbell, Michael, E-mail: mhl.campbell@gmail.com; Bergmann, Andreas, E-mail: a.bergmann@hotmail.com

2015-10-15

Vector approach often benefits optical engineers and physicists, and a vector formulation of the laws of reflection and refraction has been studied (Tkaczyk, 2012). However, the conventional reflection and refraction laws may be violated in the presence of a metasurface, and reflection and refraction at the metasurface obey generalized laws of reflection and refraction (Yu et al., 2011). In this letter, the vectorial laws of reflection and refraction at the metasurface were derived, and the matrix formulation of these vectorial laws are also obtained. These results enable highly efficient and unambiguous computations in ray-tracing problems that involve a metasurface.

Spatial orientation and electric-field-driven transport of hypericin inside of bilayer lipid membranes.

PubMed

Strejčková, Alena; Staničová, Jana; Jancura, Daniel; Miškovský, Pavol; Bánó, Gregor

2013-02-07

Fluorescence experiments were carried out to investigate the interaction of hypericin (Hyp), a natural hydrophobic photosensitizer, with artificial bilayer lipid membranes. The spatial orientation of Hyp monomers incorporated in diphytanoyl phosphatidylcholine (DPhPC) membranes was determined by measuring the dependence of the Hyp fluorescence intensity on the angle of incidence of p- and s-polarized excitation laser beams. Inside of the membrane, Hyp monomers are preferentially located in the layers near the membrane/water interface and are oriented with the S(1) ← S(0) transition dipole moments perpendicular to the membrane surface. Transport of Hyp anions between the two opposite sides of the lipid bilayer was induced by applying rectangular electric field pulses to the membrane. The characteristic time for Hyp transport through the membrane center was evaluated by the analysis of the Hyp fluorescence signal during the voltage pulses. In the zero-voltage limit, the transport time approached 70 ms and gradually decreased with higher voltage applied to the membrane. In addition, our measurements indicated an apparent pK(a) constant of 8 for Hyp deprotonation in the membrane.

Membrane Protein Mobility and Orientation Preserved in Supported Bilayers Created Directly from Cell Plasma Membrane Blebs.

PubMed

Richards, Mark J; Hsia, Chih-Yun; Singh, Rohit R; Haider, Huma; Kumpf, Julia; Kawate, Toshimitsu; Daniel, Susan

2016-03-29

Membrane protein interactions with lipids are crucial for their native biological behavior, yet traditional characterization methods are often carried out on purified protein in the absence of lipids. We present a simple method to transfer membrane proteins expressed in mammalian cells to an assay-friendly, cushioned, supported lipid bilayer platform using cell blebs as an intermediate. Cell blebs, expressing either GPI-linked yellow fluorescent proteins or neon-green fused transmembrane P2X2 receptors, were induced to rupture on glass surfaces using PEGylated lipid vesicles, which resulted in planar supported membranes with over 50% mobility for multipass transmembrane proteins and over 90% for GPI-linked proteins. Fluorescent proteins were tracked, and their diffusion in supported bilayers characterized, using single molecule tracking and moment scaling spectrum (MSS) analysis. Diffusion was characterized for individual proteins as either free or confined, revealing details of the local lipid membrane heterogeneity surrounding the protein. A particularly useful result of our bilayer formation process is the protein orientation in the supported planar bilayer. For both the GPI-linked and transmembrane proteins used here, an enzymatic assay revealed that protein orientation in the planar bilayer results in the extracellular domains facing toward the bulk, and that the dominant mode of bleb rupture is via the "parachute" mechanism. Mobility, orientation, and preservation of the native lipid environment of the proteins using cell blebs offers advantages over proteoliposome reconstitution or disrupted cell membrane preparations, which necessarily result in significant scrambling of protein orientation and typically immobilized membrane proteins in SLBs. The bleb-based bilayer platform presented here is an important step toward integrating membrane proteomic studies on chip, especially for future studies aimed at understanding fundamental effects of lipid interactions on protein activity and the roles of membrane proteins in disease pathways.

Coupling mesodomain positional ordering to intra-domain orientational ordering in block copolymer assembly

NASA Astrophysics Data System (ADS)

Burke, Christopher; Reddy, Abhiram; Prasad, Ishan; Grason, Gregory

Block copolymer (BCP) melts form a number of symmetric microphases, e.g. columnar or double gyroid phases. BCPs with a block composed of chiral monomers are observed to form bulk phases with broken chiral symmetry e.g. a phase of hexagonally ordered helical mesodomains. Other new structures may be possible, e.g. double gyroid with preferred chirality which has potential photonic applications. One approach to understanding chirality transfer from monomer to the bulk is to use self consistent field theory (SCFT) and incorporate an orientational order parameter with a preference for handed twist in chiral block segments, much like the texture of cholesteric liquid crystal. Polymer chains in achiral BCPs exhibit orientational ordering which couples to the microphase geometry; a spontaneous preference for ordering may have an effect on the geometry. The influence of a preference for chiral polar (vectorial) segment order has been studied to some extent, though the influence of coupling to chiral tensorial (nematic) order has not yet been developed. We present a computational approach using SCFT with vector and tensor order which employs well developed pseudo-spectral methods. Using this we explore how tensor order influences which structures form, and if it can promote chiral phases.

Water Dynamics in Nafion Fuel Cell Membranes: the Effects of Confinement and Structural Changes on the Hydrogen Bond Network

PubMed Central

Moilanen, David E.; Piletic, Ivan R.; Fayer, Michael D.

2008-01-01

The complex environments experienced by water molecules in the hydrophilic channels of Nafion membranes are studied by ultrafast infrared pump-probe spectroscopy. A wavelength dependent study of the vibrational lifetime of the O-D stretch of dilute HOD in H2O confined in Nafion membranes provides evidence of two distinct ensembles of water molecules. While only two ensembles are present at each level of membrane hydration studied, the characteristics of the two ensembles change as the water content of the membrane changes. Time dependent anisotropy measurements show that the orientational motions of water molecules in Nafion membranes are significantly slower than in bulk water and that lower hydration levels result in slower orientational relaxation. Initial wavelength dependent results for the anisotropy show no clear variation in the time scale for orientational motion across a broad range of frequencies. The anisotropy decay is analyzed using a model based on restricted orientational diffusion within a hydrogen bond configuration followed by total reorientation through jump diffusion. PMID:18728757

Biomechanics of the unique pterosaur pteroid

PubMed Central

Palmer, Colin; Dyke, Gareth J.

2010-01-01

Pterosaurs, flying reptiles from the Mesozoic, had wing membranes that were supported by their arm bones and a super-elongate fourth finger. Associated with the wing, pterosaurs also possessed a unique wrist bone—the pteroid—that functioned to support the forward part of the membrane in front of the leading edge, the propatagium. Pteroid shape varies across pterosaurs and reconstructions of its orientation vary (projecting anteriorly to the wing leading edge or medially, lying alongside it) and imply differences in the way that pterosaurs controlled their wings. Here we show, using biomechanical analysis and considerations of aerodynamic efficiency of a representative ornithocheirid pterosaur, that an anteriorly orientated pteroid is highly unlikely. Unless these pterosaurs only flew steadily and had very low body masses, their pteroids would have been likely to break if orientated anteriorly; the degree of movement required for a forward orientation would have introduced extreme membrane strains and required impractical tensioning in the propatagium membrane. This result can be generalized for other pterodactyloid pterosaurs because the resultant geometry of an anteriorly orientated pteroid would have reduced the aerodynamic performance of all wings and required the same impractical properties in the propatagium membrane. We demonstrate quantitatively that the more traditional reconstruction of a medially orientated pteroid was much more stable both structurally and aerodynamically, reflecting likely life position. PMID:20007183

Effects of Plant-Community Composition on the Vectorial Capacity and Fitness of the Malaria Mosquito Anopheles gambiae

PubMed Central

Stone, Christopher M.; Jackson, Bryan T.; Foster, Woodbridge A.

2012-01-01

Dynamics of Anopheles gambiae abundance and malaria transmission potential rely strongly on environmental conditions. Female and male An. gambiae use sugar and are affected by its absence, but how the presence or absence of nectariferous plants affects An. gambiae abundance and vectorial capacity has not been studied. We report on four replicates of a cohort study performed in mesocosms with sugar-poor and sugar-rich plants, in which we measured mosquito survival, biting rates, and fecundity. Survivorship was greater with access to sugar-rich plant species, and mortality patterns were age-dependent. Sugar-poor populations experienced Weibull mortality patterns, and of four populations in the sugar-rich environment, two female and three male subpopulations were better fitted by Gompertz functions. A tendency toward higher biting rates in sugar-poor mesocosms, particularly for young females, was found. Therefore, vectorial capacity was pulled in opposing directions by nectar availability, resulting in highly variable vectorial capacity values. PMID:22927493

Molecular recognition of RAS/RAF complex at the membrane: Role of RAF cysteine-rich domain

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

Travers, Timothy; Lopez Bautista, Cesar Augusto; Van, Que

Activation of RAF kinase involves the association of its RAS-binding domain (RBD) and cysteine-rich domain (CRD) with membrane-anchored RAS. However, the overall architecture of the RAS/RBD/CRD ternary complex and the orientations of its constituent domains at the membrane remain unclear. Here in this paper, we have combined all-atom and coarse-grained molecular dynamics (MD) simulations with experimental data to construct and validate a model of membrane-anchored CRD, and used this as a basis to explore models of membrane-anchored RAS/RBD/CRD complex. First, simulations of the CRD revealed that it anchors to the membrane via insertion of its two hydrophobic loops, which ismore » consistent with our NMR measurements of CRD bound to nanodiscs. Simulations of the CRD in the context of membrane-anchored RAS/RBD then show how CRD association with either RAS or RBD could play an unexpected role in guiding the membrane orientations of RAS/RBD. This finding has implications for the formation of RAS-RAS dimers, as different membrane orientations of RAS expose distinct putative dimerization interfaces.« less

Molecular recognition of RAS/RAF complex at the membrane: Role of RAF cysteine-rich domain

DOE PAGES

Travers, Timothy; Lopez Bautista, Cesar Augusto; Van, Que; ...

2018-05-31

Activation of RAF kinase involves the association of its RAS-binding domain (RBD) and cysteine-rich domain (CRD) with membrane-anchored RAS. However, the overall architecture of the RAS/RBD/CRD ternary complex and the orientations of its constituent domains at the membrane remain unclear. Here in this paper, we have combined all-atom and coarse-grained molecular dynamics (MD) simulations with experimental data to construct and validate a model of membrane-anchored CRD, and used this as a basis to explore models of membrane-anchored RAS/RBD/CRD complex. First, simulations of the CRD revealed that it anchors to the membrane via insertion of its two hydrophobic loops, which ismore » consistent with our NMR measurements of CRD bound to nanodiscs. Simulations of the CRD in the context of membrane-anchored RAS/RBD then show how CRD association with either RAS or RBD could play an unexpected role in guiding the membrane orientations of RAS/RBD. This finding has implications for the formation of RAS-RAS dimers, as different membrane orientations of RAS expose distinct putative dimerization interfaces.« less

Biochemical characterization of P-type copper ATPases

PubMed Central

Inesi, Giuseppe; Pilankatta, Rajendra; Tadini-Buoninsegni, Francesco

2014-01-01

Copper ATPases, in analogy with other members of the P-ATPase superfamily, contain a catalytic headpiece including an aspartate residue reacting with ATP to form a phosphoenzyme intermediate, and transmembrane helices containing cation-binding sites [TMBS (transmembrane metal-binding sites)] for catalytic activation and cation translocation. Following phosphoenzyme formation by utilization of ATP, bound copper undergoes displacement from the TMBS to the lumenal membrane surface, with no H+ exchange. Although PII-type ATPases sustain active transport of alkali/alkali-earth ions (i.e. Na+, Ca2+) against electrochemical gradients across defined membranes, PIB-type ATPases transfer transition metal ions (i.e. Cu+) from delivery to acceptor proteins and, prominently in mammalian cells, undergo trafficking from/to various membrane compartments. A specific component of copper ATPases is the NMBD (N-terminal metal-binding domain), containing up to six copper-binding sites in mammalian (ATP7A and ATP7B) enzymes. Copper occupancy of NMBD sites and interaction with the ATPase headpiece are required for catalytic activation. Furthermore, in the presence of copper, the NMBD allows interaction with protein kinase D, yielding phosphorylation of serine residues, ATP7B trafficking and protection from proteasome degradation. A specific feature of ATP7A is glycosylation and stabilization on plasma membranes. Cisplatin, a platinum-containing anti-cancer drug, binds to copper sites of ATP7A and ATP7B, and undergoes vectorial displacement in analogy with copper. PMID:25242165

[Chagas's disease and deep ecology: the anti-vectorial fight in question].

PubMed

Siqueira-Batista, Rodrigo; Gomes, Andréia Patrícia; Rôças, Giselle; Cotta, Rosângela Minardi Mitre; Rubião, Eduardo Cárdenas Nogueira; Pissinatti, Alcides

2011-02-01

The inter-relations between man and the environment are among the main themes currently debated by the Brazilian public health. On such horizon, the questions concerning Chagas's disease are found to remain specially in the scope of the directed actions of control to the triatomine, the anti-vectorial fight , though already a century since its first description by Carlos Chagas, a major epidemiological problem in Latin America. Based on these considerations the present article will seek to discuss the main ecological aspects related to the American trypanosomiasis, emphasizing the control of the vectorial transmission in the context of the deep ecology.

Fully vectorial accelerating diffraction-free Helmholtz beams.

PubMed

Aleahmad, Parinaz; Miri, Mohammad-Ali; Mills, Matthew S; Kaminer, Ido; Segev, Mordechai; Christodoulides, Demetrios N

2012-11-16

We show that new families of diffraction-free nonparaxial accelerating optical beams can be generated by considering the symmetries of the underlying vectorial Helmholtz equation. Both two-dimensional transverse electric and magnetic accelerating wave fronts are possible, capable of moving along elliptic trajectories. Experimental results corroborate these predictions when these waves are launched from either the major or minor axis of the ellipse. In addition, three-dimensional spherical nondiffracting field configurations are presented along with their evolution dynamics. Finally, fully vectorial self-similar accelerating optical wave solutions are obtained via oblate-prolate spheroidal wave functions. In all occasions, these effects are illustrated via pertinent examples.

MreB filaments align along greatest principal membrane curvature to orient cell wall synthesis

PubMed Central

Szwedziak, Piotr; Wong, Felix; Schaefer, Kaitlin; Izoré, Thierry; Renner, Lars D; Holmes, Matthew J; Sun, Yingjie; Bisson-Filho, Alexandre W; Walker, Suzanne; Amir, Ariel; Löwe, Jan

2018-01-01

MreB is essential for rod shape in many bacteria. Membrane-associated MreB filaments move around the rod circumference, helping to insert cell wall in the radial direction to reinforce rod shape. To understand how oriented MreB motion arises, we altered the shape of Bacillus subtilis. MreB motion is isotropic in round cells, and orientation is restored when rod shape is externally imposed. Stationary filaments orient within protoplasts, and purified MreB tubulates liposomes in vitro, orienting within tubes. Together, this demonstrates MreB orients along the greatest principal membrane curvature, a conclusion supported with biophysical modeling. We observed that spherical cells regenerate into rods in a local, self-reinforcing manner: rapidly propagating rods emerge from small bulges, exhibiting oriented MreB motion. We propose that the coupling of MreB filament alignment to shape-reinforcing peptidoglycan synthesis creates a locally-acting, self-organizing mechanism allowing the rapid establishment and stable maintenance of emergent rod shape. PMID:29469806

Molecular density functional theory of water including density-polarization coupling.

PubMed

Jeanmairet, Guillaume; Levy, Nicolas; Levesque, Maximilien; Borgis, Daniel

2016-06-22

We present a three-dimensional molecular density functional theory of water derived from first-principles that relies on the particle's density and multipolar polarization density and includes the density-polarization coupling. This brings two main benefits: (i) scalar density and vectorial multipolar polarization density fields are much more tractable and give more physical insight than the full position and orientation densities, and (ii) it includes the full density-polarization coupling of water, that is known to be non-vanishing but has never been taken into account. Furthermore, the theory requires only the partial charge distribution of a water molecule and three measurable bulk properties, namely the structure factor and the Fourier components of the longitudinal and transverse dielectric susceptibilities.

Actin protofilament orientation in deformation of the erythrocyte membrane skeleton.

PubMed Central

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

2000-01-01

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

Cellular Mechanisms of Gravitropic Response in Higher Plants

NASA Astrophysics Data System (ADS)

Medvedev, Sergei; Smolikova, Galina; Pozhvanov, Gregory; Suslov, Dmitry

The evolutionary success of land plants in adaptation to the vectorial environmental factors was based mainly on the development of polarity systems. In result, normal plant ontogenesis is based on the positional information. Polarity is a tool by which the developing plant organs and tissues are mapped and the specific three-dimensional structure of the organism is created. It is due to their polar organization plants are able to orient themselves relative to the gravity vector and different vectorial cues, and to respond adequately to various stimuli. Gravitation is one of the most important polarized environmental factor that guides the development of plant organisms in space. Every plant can "estimate" its position relative to the gravity vector and correct it, if necessary, by means of polarized growth. The direction and the magnitude of gravitational stimulus are constant during the whole plant ontogenesis. The key plant response to the action of gravity is gravitropism, i.e. the directed growth of organs with respect to the gravity vector. This response is a very convenient model to study the mechanisms of plant orientation in space. The present report is focused on the main cellular mechanisms responsible for graviropic bending in higher plants. These mechanisms and structures include electric polarization of plant cells, Ca ({2+) }gradients, cytoskeleton, G-proteins, phosphoinositides and the machinery responsible for asymmetric auxin distribution. Those mechanisms tightly interact demonstrating some hierarchy and multiple feedbacks. The Ca (2+) gradients provide the primary physiological basis of polarity in plant cells. Calcium ions influence on the bioelectric potentials, the organization of actin cytoskeleton, the activity of Ca (2+) -binding proteins and Ca (2+) -dependent protein kinases. Protein kinases modulate transcription factors activity thereby regulating the gene expression and switching the developmental programs. Actin cytoskeleton affects the molecular machinery of polar auxin transport. It results in the changes of auxin gradients in plant organs and tissues, which modulate all cellular mechanisms of polarity via multiple feedback loops. The understanding of the mechanisms of plant organism orientation relative to the gravity vector will allow us to develop efficient technologies for plant growing in microgravity conditions at orbital space stations and during long piloted space flights. This work was supported by the grant of Russian Foundation for Basic Research (N 14-04-01-624) and by the grant of St.-Petersburg State University (N 1.38.233.2014).

Control of gravitropic orientation. I. Non-vertical orientation by primary roots of maize results from decay of competence for orthogravitropic induction

NASA Technical Reports Server (NTRS)

LaMotte, Clifford E.; Pickard, Barbara G.

2004-01-01

Plant organs may respond to gravity by vertical (orthogravitropic), oblique (plagiogravitropic) or horizontal (diagravitropic) growth. Primary roots of maize (Zea mays L.) provide a good system for studying such behaviours because they are reportedly capable of displaying all three responses. In current work using maize seedlings of the Silver Queen cultivar, stabilisation of growth at an oblique orientation was commonplace. Hypothetically, plagiogravitropism may be accomplished either by a process we call graded orthogravitropism or by hunting about a sensed non-vertical setpoint. In graded orthotropism primary bending is unidirectional and depends on facilitative stimuli that determine its extent. The hallmark of the setpoint mechanism is restorative curvature of either sign following a displacement; both diagravitropism and orthogravitropism are based on setpoints. Roots settled in a plagiogravitropic orientation were tested with various illumination and displacement protocols designed to distinguish between these two hypotheses. The tests refuted the setpoint hypothesis and supported that of graded orthotropism. No evidence of diagravitropism could be found, thus, earlier claims were likely based on inadequately controlled observations of graded orthotropism. We propose that orthotropism is graded by the sequential action of dual gravity receptors: induction of a vectorial gravitropic response requires gravitational induction of a separate facilitative response, whose decay in the absence of fresh stimuli can brake gravitropism at plagiotropic angles.

Crystalline Membranes

NASA Technical Reports Server (NTRS)

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

2008-01-01

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

Mechanisms of charge transfer in human copper ATPases ATP7A and ATP7B.

PubMed

Tadini-Buoninsegni, Francesco; Smeazzetto, Serena

2017-04-01

ATP7A and ATP7B are Cu + -transporting ATPases of subclass IB and play a fundamental role in intracellular copper homeostasis. ATP7A/B transfer Cu + ions across the membrane from delivery to acceptor proteins without establishing a free Cu + gradient. Transfer of copper across the membrane is coupled to ATP hydrolysis. Current measurements on solid supported membranes (SSM) were performed to investigate the mechanism of copper-related charge transfer across ATP7A and ATP7B. SSM measurements demonstrated that electrogenic copper displacement occurs within ATP7A/B following addition of ATP and formation of the phosphorylated intermediate. Comparison of the time constants for cation displacement in ATP7A/B and sarcoplasmic reticulum Ca 2+ -ATPase is consistent with the slower phosphoenzyme formation in copper ATPases. Moreover, ATP-dependent copper transfer in ATP7A/B is not affected by varying the pH, suggesting that net proton counter-transport may not occur in copper ATPases. Platinum anticancer drugs activate ATP7A/B and are subjected to ATP-dependent vectorial displacement with a mechanism analogous to that of copper. © 2016 IUBMB Life, 69(4):218-225, 2017. © 2017 International Union of Biochemistry and Molecular Biology.

The Heteronuclear Single-Quantum Correlation (HSQC) Experiment: Vectors versus Product Operators

ERIC Educational Resources Information Center

de la Vega-Herna´ndez, Karen; Antuch, Manuel

2015-01-01

A vectorial representation of the full sequence of events occurring during the 2D-NMR heteronuclear single-quantum correlation (HSQC) experiment is presented. The proposed vectorial representation conveys an understanding of the magnetization evolution during the HSQC pulse sequence for those who have little or no quantum mechanical background.…

Vectorial finite elements for solving the radiative transfer equation

NASA Astrophysics Data System (ADS)

Badri, M. A.; Jolivet, P.; Rousseau, B.; Le Corre, S.; Digonnet, H.; Favennec, Y.

2018-06-01

The discrete ordinate method coupled with the finite element method is often used for the spatio-angular discretization of the radiative transfer equation. In this paper we attempt to improve upon such a discretization technique. Instead of using standard finite elements, we reformulate the radiative transfer equation using vectorial finite elements. In comparison to standard finite elements, this reformulation yields faster timings for the linear system assemblies, as well as for the solution phase when using scattering media. The proposed vectorial finite element discretization for solving the radiative transfer equation is cross-validated against a benchmark problem available in literature. In addition, we have used the method of manufactured solutions to verify the order of accuracy for our discretization technique within different absorbing, scattering, and emitting media. For solving large problems of radiation on parallel computers, the vectorial finite element method is parallelized using domain decomposition. The proposed domain decomposition method scales on large number of processes, and its performance is unaffected by the changes in optical thickness of the medium. Our parallel solver is used to solve a large scale radiative transfer problem of the Kelvin-cell radiation.

Vectorial structures of linear-polarized Butterfly-Gauss vortex beams in the far zone

NASA Astrophysics Data System (ADS)

Cheng, Ke; Zhou, Yan; Lu, Gang; Yao, Na; Zhong, Xianqiong

2018-05-01

By introducing the Butterfly catastrophe to optics, the far-zone vectorial structures of Butterfly-Gauss beam with vortex and non-vortex are studied using the angular spectrum representation and stationary phase method. The influence of topological charge, linear-polarized angle, off-axis distance and scaling length on the far-zone vectorial structures, especially in the Poynting vector and angular momentum density of the corresponding beam is emphasized. The results show that the embedded optical vortex at source plane lead to special dark zones in the far zone, where the number of dark zone equals the absolute value of topological charge of optical vortex. Furthermore, the symmetry and direction of the special dark zones can be controlled by off-axis distance and scaling length, respectively. The linear-polarized angle adjusts only the Poynting vectors of TE and TM terms, but it does not affect those of whole beam. Finally, the vectorial expressions also indicate that the total angular momentum density is certainly zero owing to the far-zone stable structures rather than rotation behaviors.

Cellulose microfibrils: visualization of biosynthetic and orienting complexes in association with the plasma membrane.

PubMed

Brown, R M; Montezinos, D

1976-01-01

Cellulose microfibril biosynthesis, assembly, and orientation in the unicellular green alga, Oocystis, is visualized in association with a linear enzyme complex embedded in the B face of the plasma membrane. Granule bands of the A face and complementary ridges of the B face are postulated to assist in the orientation of recently synthesized microfibrils. A model for microfibril synthesis and orientation is proposed and correlated with current hypotheses regarding cellulose biosynthesis in higher plants.

Changes in the anisotropy of oriented membrane dynamics induced by myelin basic protein

NASA Astrophysics Data System (ADS)

Natali, F.; Gliozzi, A.; Rolandi, R.; Relini, A.; Cavatorta, P.; Deriu, A.; Fasano, A.; Riccio, P.

We report recent results showing the evidence of the effect induced by physiological amounts of myelin basic protein (MBP) on the dynamics of dimyristoyl L-a-phosphatidic acid (DMPA) membranes. Incoherent elastic neutron scattering scans, performed over a wide temperature range, have shown that the anisotropy of motions in oriented membranes is significantly enhanced by the presence of MBP.

An elaborated feeding cycle model for reductions in vectorial capacity of night-biting mosquitoes by insecticide-treated nets.

PubMed

Le Menach, Arnaud; Takala, Shannon; McKenzie, F Ellis; Perisse, Andre; Harris, Anthony; Flahault, Antoine; Smith, David L

2007-01-25

Insecticide Treated Nets (ITNs) are an important tool for malaria control. ITNs are effective because they work on several parts of the mosquito feeding cycle, including both adult killing and repelling effects. Using an elaborated description of the classic feeding cycle model, simple formulas have been derived to describe how ITNs change mosquito behaviour and the intensity of malaria transmission, as summarized by vectorial capacity and EIR. The predicted changes are illustrated as a function of the frequency of ITN use for four different vector populations using parameter estimates from the literature. The model demonstrates that ITNs simultaneously reduce mosquitoes' lifespans, lengthen the feeding cycle, and by discouraging human biting divert more bites onto non-human hosts. ITNs can substantially reduce vectorial capacity through small changes to all of these quantities. The total reductions in vectorial capacity differ, moreover, depending on baseline behavior in the absence of ITNs. Reductions in lifespan and vectorial capacity are strongest for vector species with high baseline survival. Anthropophilic and zoophilic species are affected differently by ITNs; the feeding cycle is lengthened more for anthrophilic species, and the proportion of bites that are diverted onto non-human hosts is higher for zoophilic species. This model suggests that the efficacy of ITNs should be measured as a total reduction in transmission intensity, and that the quantitative effects will differ by species and by transmission intensity. At very high rates of ITN use, ITNs can generate large reductions in transmission intensity that could provide very large reductions in transmission intensity, and effective malaria control in some areas, especially when used in combination with other control measures. At high EIR, ITNs will probably not substantially reduce the parasite rate, but when transmission intensity is low, reductions in vectorial capacity combine with reductions in the parasite rate to generate very large reductions in EIR.

The orientation of iron–sulphur clusters in membrane multilayers prepared from aerobically-grown Escherichia coli K12 and a cytochrome-deficient mutant

PubMed Central

Blum, Haywood; Poole, Robert K.; Ohnishi, Tomoko

1980-01-01

1. Membrane particles prepared from ultrasonically-disrupted, aerobically-grown Escherichia coli were centrifuged on to a plastic film that was supported perpendicular to the centrifugal field to yield oriented membrane multilayers. In such preparations, there is a high degree of orientation of the planes of the membranes such that they lie parallel to each other and to the supporting film. 2. When dithionite- or succinate-reduced multilayers are rotated in the magnetic field of an e.p.r. spectrometer, about an axis lying in the membrane plane, angular-dependent signals from an iron–sulphur cluster at gx=1.92, gy=1.93 and gz=2.02 are seen. The g=1.93 signal has maximal amplitude when the plane of the multilayer is perpendicular to the magnetic field. Conversely, the g=2.02 signal is maximal when the plane of the multilayer is parallel with the magnetic field. 3. Computer simulations of the experimental data show that the cluster lies in the cytoplasmic membrane with the gy axis perpendicular to the membrane plane and with the gx and gz axes lying in the membrane plane. 4. In partially-oxidized multilayers, a signal resembling the mitochondrial high-potential iron–sulphur protein (Hipip) is seen whose gz=2.02 axis may be deduced as lying perpendicular to the membrane plane. 5. Appropriate choice of sample temperature and receiver gain reveals two further signals in partially-reduced multilayers: a g=2.09 signal arises from a cluster with its gz axis in the membrane plane, whereas a g=2.04 signal is from a cluster with the gz axis lying along the membrane normal. 6. Membrane particles from a glucose-grown, haem-deficient mutant contain dramatically-lowered levels of cytochromes and exhibit, in addition to the iron–sulphur clusters seen in the parental strain, a major signal at g=1.90. 7. Only the latter may be demonstrated to be oriented in multilayer preparations from the mutant. 8. Comparisons are drawn between the orientations of the iron–sulphur proteins in the cytoplasmic membrane of E. coli and those in mitochondrial membranes. The effects of diminished cytochrome content on the properties of the iron–sulphur proteins are discussed. PMID:6258566

Emergence of polysaccharide membrane walls through macro-space partitioning via interfacial instability.

PubMed

Okeyoshi, Kosuke; Okajima, Maiko K; Kaneko, Tatsuo

2017-07-21

Living organisms in drying environments build anisotropic structures and exhibit directionality through self-organization of biopolymers. However, the process of macro-scale assembly is still unknown. Here, we introduce a dissipative structure through a non-equilibrium process between hydration and deposition in the drying of a polysaccharide liquid crystalline solution. By controlling the geometries of the evaporation front in a limited space, multiple nuclei emerge to grow vertical membrane walls with macroscopic orientation. Notably, the membranes are formed through rational orientation of rod-like microassemblies along the dynamic three-phase contact line. Additionally, in the non-equilibrium state, a dissipative structure is ultimately immobilized as a macroscopically partitioned space by multiple vertical membranes. We foresee that such oriented membranes will be applicable to soft biomaterials with direction controllability, and the macroscopic space partitionings will aid in the understanding of the space recognition ability of natural products under drying environments.

An Acidic Cluster in the Cytosolic Domain of Human Cytomegalovirus Glycoprotein B Is a Signal for Endocytosis from the Plasma Membrane

PubMed Central

Tugizov, Sharof; Maidji, Ekaterina; Xiao, Jianqiao; Pereira, Lenore

1999-01-01

We previously reported that human cytomegalovirus (CMV) glycoprotein B (gB) is transported to apical membranes in CMV-infected polarized retinal pigment epithelial (ARPE-19) cells and in Madin-Darby canine kidney (MDCK) epithelial cells constitutively expressing gB. The cytosolic domain of gB contains a cluster of acidic amino acids, a motif that plays a pivotal role in vectorial trafficking in polarized epithelial cells and may also function as a signal for entry into the endocytic pathway. Here we compared gB internalization and recycling to the plasma membrane in CMV-infected human fibroblasts (HF) and ARPE-19 cells by using antibody-internalization experiments. Immunofluorescence and quantitative assays showed that gB was internalized from the cell surface into clathrin-coated transport vesicles and then recycled to the plasma membrane. gB colocalized with clathrin-coated vesicles containing the transferrin receptor in the early endocytic/recycling pathway, indicating that gB traffics in this pathway. The specific role of the acidic cluster in regulating the sorting of gB-containing vesicles in the early endocytic/recycling pathway was examined in MDCK cells expressing mutated gB derivatives. Immunofluorescence assays showed that derivatives lacking the acidic cluster were impaired in internalization and failed to recycle. These findings, together with our earlier observation that the acidic cluster is a key determinant for targeting gB molecules to apical membranes in epithelial cells, establish that this signal is recognized by cellular proteins that participate in polarized sorting and transport in the early endocytic/recycling pathway. PMID:10482621

Prediction of protein orientation upon immobilization on biological and nonbiological surfaces

NASA Astrophysics Data System (ADS)

Talasaz, Amirali H.; Nemat-Gorgani, Mohsen; Liu, Yang; Ståhl, Patrik; Dutton, Robert W.; Ronaghi, Mostafa; Davis, Ronald W.

2006-10-01

We report on a rapid simulation method for predicting protein orientation on a surface based on electrostatic interactions. New methods for predicting protein immobilization are needed because of the increasing use of biosensors and protein microarrays, two technologies that use protein immobilization onto a solid support, and because the orientation of an immobilized protein is important for its function. The proposed simulation model is based on the premise that the protein interacts with the electric field generated by the surface, and this interaction defines the orientation of attachment. Results of this model are in agreement with experimental observations of immobilization of mitochondrial creatine kinase and type I hexokinase on biological membranes. The advantages of our method are that it can be applied to any protein with a known structure; it does not require modeling of the surface at atomic resolution and can be run relatively quickly on readily available computing resources. Finally, we also propose an orientation of membrane-bound cytochrome c, a protein for which the membrane orientation has not been unequivocally determined. electric double layer | electrostatic simulations | orientation flexibility

Biologically Complex Planar Cell Plasma Membranes Supported on Polyelectrolyte Cushions Enhance Transmembrane Protein Mobility and Retain Native Orientation.

PubMed

Liu, Han-Yuan; Chen, Wei-Liang; Ober, Christopher K; Daniel, Susan

2018-01-23

Reconstituted supported lipid bilayers (SLB) are widely used as in vitro cell-surface models because they are compatible with a variety of surface-based analytical techniques. However, one of the challenges of using SLBs as a model of the cell surface is the limited complexity in membrane composition, including the incorporation of transmembrane proteins and lipid diversity that may impact the activity of those proteins. Additionally, it is challenging to preserve the transmembrane protein native orientation, function, and mobility in SLBs. Here, we leverage the interaction between cell plasma membrane vesicles and polyelectrolyte brushes to create planar bilayers from cell plasma membrane vesicles that have budded from the cell surface. This approach promotes the direct incorporation of membrane proteins and other species into the planar bilayer without using detergent or reconstitution and preserves membrane constituents. Furthermore, the structure of the polyelectrolyte brush serves as a cushion between the planar bilayer and rigid supporting surface, limiting the interaction of the cytosolic domains of membrane proteins with this surface. Single particle tracking was used to analyze the motion of GPI-linked yellow fluorescent proteins (GPI-YFP) and neon-green fused transmembrane P2X2 receptors (P2X2-neon) and shows that this platform retains over 75% mobility of multipass transmembrane proteins in its native membrane environment. An enzyme accessibility assay confirmed that the protein orientation is preserved and results in the extracellular domain facing toward the bulk phase and the cytosolic side facing the support. Because the platform presented here retains the complexity of the cell plasma membrane and preserves protein orientation and mobility, it is a better representative mimic of native cell surfaces, which may find many applications in biological assays aimed at understanding cell membrane phenomena.

Magnetic heading reference

NASA Technical Reports Server (NTRS)

Garner, H. D. (Inventor)

1983-01-01

Devices are disclosed for vectorially summing two signals. In a first embodiment, the vectorial summation is implemented by a mechanical sin/cos mechanism in which a crank drives two linear potentiometers out of phase. In a second embodiment, a polarized light resolver generates the sin and cos functions. In a third embodiment, a printed circuit resolver generates the sin and cos functions.

Membrane Orientation of Gαiβ1γ2 and Gβ1 γ2 Determined via Combined Vibrational Spectroscopic Studies

PubMed Central

Yang, Pei; Boughton, Andrew; Homan, Kristoff T.; Tesmer, John J.G.; Chen, Zhan

2013-01-01

The manner in which the heterotrimeric G protein complexes Gβ1γ2 and Gαiβ1γ2 interact with membranes is likely related to their biological function. We combined complementary measurements from sum frequency generation (SFG) vibrational and attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy to determine the possible membrane orientations of Gβ1γ2 and the Gαiβ1γ2 heterotrimer more precisely than could be achieved using SFG alone. The most likely orientations of Gβ1γ2 and the Gαiβ1γ2 heterotrimer were both determined to fall within a similar narrow range of twist and tilt angles, suggesting that Gβ1γ2 may bind to Gαi without a significant change in orientation. This “basal” orientation seems to depend primarily on the geranylgeranylated C-terminus of Gγ2 along with basic residues at the N-terminus of Gαi, and suggests that activated G protein-coupled receptors (GPCRs) must reorient G protein heterotrimers at lipid bilayers to catalyze nucleotide exchange. The innovative methodologies developed in this paper can be widely applied to study the membrane orientation of other proteins in situ. PMID:23461393

Final Technical Report ONT/ASEE (Office of Naval Technology/American Society for Engineering Education) Postdoctoral Fellow on Contract N00014-83-D-0689.

DTIC Science & Technology

1988-02-19

phosphatase with BCIP/NBT appears to produce the most clearly visible spot on the membrane. The spot is a purple-blue color, but one must orient the membrane...phosphatase-labeled antibodies. With any of these reagents, we can produce a discrete and clearly visible spot on nitrocellulose membranes. The intensity of...substrate systems are used; however, one has to orient the membrane towards the light in such a manner that the spot is clearly visible . We have found

Effects of Carbon Nanotubes in Barrier Epithelial Cells via Effects on Lipid Bilayers

NASA Astrophysics Data System (ADS)

Lewis, Shanta

Carbon nanotubes (CNTs) are one of the most common nanoparticles (NP) found in workplace air. Therefore, there is a strong chance that these NP will enter the human body. They have similar physical properties to asbestos, a known toxic material, yet there is limited evidence showing that CNTs may be hazardous to human barrier epithelia. In previous studies done in our laboratory, the effects of CNTs on the barrier function in the human airway epithelial cell line (Calu-3) were measured. Measurements were done using electrophysiology, a technique which measures both transepithelial electrical resistance (TEER), a measure of monolayer integrity, and short circuit current (SCC) which is a measure of vectorial ion transport across the cell monolayer. The research findings showed that select physiologically relevant concentrations of long single-wall (SW) and multi-wall (MW) CNTs significantly decreased the stimulated SCC of the Calu-3 cells compared to untreated cultures. Calu-3 cells showed decreases in TEER when incubated for 48 hours (h) with concentrations of MWCNT ranging from 4microg/cm2 to 0.4ng/cm2 and SWCNT ranging from 4microg/cm2 to 0.04ng/cm2. The impaired cellular function, despite sustained cell viability, led us to investigate the mechanism by which the CNTs were affecting the cell membrane. We investigated the interaction of short MWCNTs with model lipid membranes using an ion channel amplifier, Planar Bilayer Workstation. Membranes were synthesized using neutral diphytanoylphosphatidylcholine (DPhPC) and negatively charged diphytanoylphosphatidylserine (DPhPS) lipids. Gramicidin A (GA), an ion channel reporter protein, was used to measure changes in ion channel conductance due to CNT exposures. Synthetic membranes exposed to CNTs allowed bursts of currents to cross the membrane when they were added to the membrane buffer system. When added to the membrane in the presence of GA, they distorted channel formation and reduced membrane stability.

Invisible liposomes: refractive index matching with sucrose enables flow dichroism assessment of peptide orientation in lipid vesicle membrane.

PubMed

Ardhammar, Malin; Lincoln, Per; Nordén, Bengt

2002-11-26

Valuable information on protein-membrane organization may in principle be obtained from polarized-light absorption (linear dichroism, LD) measurement on shear-aligned lipid vesicle bilayers as model membranes. However, attempts to probe LD in the UV wavelength region (<250 nm) have so far failed because of strong polarized light scattering from the vesicles. Using sucrose to match the refractive index and suppress the light scattering of phosphatidylcholine vesicles, we have been able to detect LD bands also in the peptide-absorbing region (200-230 nm). The potential of refractive index matching in vesicle LD as a general method for studying membrane protein structure was investigated for the membrane pore-forming oligopeptide gramicidin incorporated into the liposome membranes. In the presence of sucrose, the LD signals arising from oriented tryptophan side chains as well as from n-->pi* and pi-->pi* transitions of the amide chromophore of the polypeptide backbone could be studied. The observation of a strongly negative LD for the first exciton transition ( approximately 204 nm) is consistent with a membrane-spanning orientation of two intertwined parallel gramicidin helices, as predicted by coupled-oscillator theory.

Structure and Orientation of a Voltage-Sensor Toxin in Lipid Membranes

PubMed Central

Jung, Hyun Ho; Jung, Hoi Jong; Milescu, Mirela; Lee, Chul Won; Lee, Seungkyu; Lee, Ju Yeon; Eu, Young-Jae; Kim, Ha Hyung; Swartz, Kenton J.; Kim, Jae Il

2010-01-01

Abstract Amphipathic protein toxins from tarantula venom inhibit voltage-activated potassium (Kv) channels by binding to a critical helix-turn-helix motif termed the voltage sensor paddle. Although these toxins partition into membranes to bind the paddle motif, their structure and orientation within the membrane are unknown. We investigated the interaction of a tarantula toxin named SGTx with membranes using both fluorescence and NMR spectroscopy. Depth-dependent fluorescence-quenching experiments with brominated lipids suggest that Trp30 in SGTx is positioned ∼9 Å from the center of the bilayer. NMR spectra reveal that the inhibitor cystine knot structure of the toxin does not radically change upon membrane partitioning. Transferred cross-saturation NMR experiments indicate that the toxin's hydrophobic protrusion contacts the hydrophobic core of the membrane, whereas most surrounding polar residues remain at interfacial regions of the bilayer. The inferred orientation of the toxin reveals a twofold symmetry in the arrangement of basic and hydrophobic residues, a feature that is conserved among tarantula toxins. These results have important implications for regions of the toxin involved in recognizing membranes and voltage-sensor paddles, and for the mechanisms by which tarantula toxins alter the activity of different types of ion channels. PMID:20643084

“Highly evolvable malaria vectors: the genomes of 16 Anopheles mosquitoes”

PubMed Central

Neafsey, Daniel E.; Waterhouse, Robert M.; Abai, Mohammad R.; Aganezov, Sergey S.; Alekseyev, Max A.; Allen, James E.; Amon, James; Arcà, Bruno; Arensburger, Peter; Artemov, Gleb; Assour, Lauren A.; Basseri, Hamidreza; Berlin, Aaron; Birren, Bruce W.; Blandin, Stephanie A.; Brockman, Andrew I.; Burkot, Thomas R.; Burt, Austin; Chan, Clara S.; Chauve, Cedric; Chiu, Joanna C.; Christensen, Mikkel; Costantini, Carlo; Davidson, Victoria L.M.; Deligianni, Elena; Dottorini, Tania; Dritsou, Vicky; Gabriel, Stacey B.; Guelbeogo, Wamdaogo M.; Hall, Andrew B.; Han, Mira V.; Hlaing, Thaung; Hughes, Daniel S.T.; Jenkins, Adam M.; Jiang, Xiaofang; Jungreis, Irwin; Kakani, Evdoxia G.; Kamali, Maryam; Kemppainen, Petri; Kennedy, Ryan C.; Kirmitzoglou, Ioannis K.; Koekemoer, Lizette L.; Laban, Njoroge; Langridge, Nicholas; Lawniczak, Mara K.N.; Lirakis, Manolis; Lobo, Neil F.; Lowy, Ernesto; MacCallum, Robert M.; Mao, Chunhong; Maslen, Gareth; Mbogo, Charles; McCarthy, Jenny; Michel, Kristin; Mitchell, Sara N.; Moore, Wendy; Murphy, Katherine A.; Naumenko, Anastasia N.; Nolan, Tony; Novoa, Eva M.; O'Loughlin, Samantha; Oringanje, Chioma; Oshaghi, Mohammad A.; Pakpour, Nazzy; Papathanos, Philippos A.; Peery, Ashley N.; Povelones, Michael; Prakash, Anil; Price, David P.; Rajaraman, Ashok; Reimer, Lisa J.; Rinker, David C.; Rokas, Antonis; Russell, Tanya L.; Sagnon, N'Fale; Sharakhova, Maria V.; Shea, Terrance; Simão, Felipe A.; Simard, Frederic; Slotman, Michel A.; Somboon, Pradya; Stegniy, Vladimir; Struchiner, Claudio J.; Thomas, Gregg W.C.; Tojo, Marta; Topalis, Pantelis; Tubio, José M.C.; Unger, Maria F.; Vontas, John; Walton, Catherine; Wilding, Craig S.; Willis, Judith H.; Wu, Yi-Chieh; Yan, Guiyun; Zdobnov, Evgeny M.; Zhou, Xiaofan; Catteruccia, Flaminia; Christophides, George K.; Collins, Frank H.; Cornman, Robert S.; Crisanti, Andrea; Donnelly, Martin J.; Emrich, Scott J.; Fontaine, Michael C.; Gelbart, William; Hahn, Matthew W.; Hansen, Immo A.; Howell, Paul I.; Kafatos, Fotis C.; Kellis, Manolis; Lawson, Daniel; Louis, Christos; Luckhart, Shirley; Muskavitch, Marc A.T.; Ribeiro, José M.; Riehle, Michael A.; Sharakhov, Igor V.; Tu, Zhijian; Zwiebel, Laurence J.; Besansky, Nora J.

2015-01-01

Variation in vectorial capacity for human malaria among Anopheles mosquito species is determined by many factors, including behavior, immunity, and life history. To investigate the genomic basis of vectorial capacity and explore new avenues for vector control, we sequenced the genomes of 16 anopheline mosquito species from diverse locations spanning ~100 million years of evolution. Comparative analyses show faster rates of gene gain and loss, elevated gene shuffling on the X chromosome, and more intron losses, relative to Drosophila. Some determinants of vectorial capacity, such as chemosensory genes, do not show elevated turnover, but instead diversify through protein-sequence changes. This dynamism of anopheline genes and genomes may contribute to their flexible capacity to take advantage of new ecological niches, including adapting to humans as primary hosts. PMID:25554792

Molecular dynamics simulations on PGLa using NMR orientational constraints.

PubMed

Sternberg, Ulrich; Witter, Raiker

2015-11-01

NMR data obtained by solid state NMR from anisotropic samples are used as orientational constraints in molecular dynamics simulations for determining the structure and dynamics of the PGLa peptide within a membrane environment. For the simulation the recently developed molecular dynamics with orientational constraints technique (MDOC) is used. This method introduces orientation dependent pseudo-forces into the COSMOS-NMR force field. Acting during a molecular dynamics simulation these forces drive molecular rotations, re-orientations and folding in such a way that the motional time-averages of the tensorial NMR properties are consistent with the experimentally measured NMR parameters. This MDOC strategy does not depend on the initial choice of atomic coordinates, and is in principle suitable for any flexible and mobile kind of molecule; and it is of course possible to account for flexible parts of peptides or their side-chains. MDOC has been applied to the antimicrobial peptide PGLa and a related dimer model. With these simulations it was possible to reproduce most NMR parameters within the experimental error bounds. The alignment, conformation and order parameters of the membrane-bound molecule and its dimer were directly derived with MDOC from the NMR data. Furthermore, this new approach yielded for the first time the distribution of segmental orientations with respect to the membrane and the order parameter tensors of the dimer systems. It was demonstrated the deuterium splittings measured at the peptide to lipid ratio of 1/50 are consistent with a membrane spanning orientation of the peptide.

Cholesterol orientation and tilt modulus in DMPC bilayers

PubMed Central

Khelashvili, George; Pabst, Georg; Harries, Daniel

2010-01-01

We performed molecular dynamics (MD) simulations of hydrated bilayers containing mixtures of dimyristoylphosphatidylcholine (DMPC) and Cholesterol at various ratios, to study the effect of cholesterol concentration on its orientation, and to characterize the link between cholesterol tilt and overall phospholipid membrane organization. The simulations show a substantial probability for cholesterol molecules to transiently orient perpendicular to the bilayer normal, and suggest that cholesterol tilt may be an important factor for inducing membrane ordering. In particular, we find that as cholesterol concentration increases (1%–40% cholesterol) the average cholesterol orientation changes in a manner strongly (anti)correlated with the variation in membrane thickness. Furthermore, cholesterol orientation is found to be determined by the aligning force exerted by other cholesterol molecules. To quantify this aligning field, we analyzed cholesterol orientation using, to our knowledge, the first estimates of the cholesterol tilt modulus χ from MD simulations. Our calculations suggest that the aligning field that determines χ is indeed strongly linked to sterol composition. This empirical parameter (χ) should therefore become a useful quantitative measure to describe cholesterol interaction with other lipids in bilayers, particularly in various coarse-grained force fields. PMID:20518573

A Novel Domain Assembly Routine for Creating Full-Length Models of Membrane Proteins from Known Domain Structures.

PubMed

Koehler Leman, Julia; Bonneau, Richard

2018-04-03

Membrane proteins composed of soluble and membrane domains are often studied one domain at a time. However, to understand the biological function of entire protein systems and their interactions with each other and drugs, knowledge of full-length structures or models is required. Although few computational methods exist that could potentially be used to model full-length constructs of membrane proteins, none of these methods are perfectly suited for the problem at hand. Existing methods require an interface or knowledge of the relative orientations of the domains or are not designed for domain assembly, and none of them are developed for membrane proteins. Here we describe the first domain assembly protocol specifically designed for membrane proteins that assembles intra- and extracellular soluble domains and the transmembrane domain into models of the full-length membrane protein. Our protocol does not require an interface between the domains and samples possible domain orientations based on backbone dihedrals in the flexible linker regions, created via fragment insertion, while keeping the transmembrane domain fixed in the membrane. For five examples tested, our method mp_domain_assembly, implemented in RosettaMP, samples domain orientations close to the known structure and is best used in conjunction with experimental data to reduce the conformational search space.

Cavity hydration dynamics in cytochrome c oxidase and functional implications

PubMed Central

Son, Chang Yun; Cui, Qiang

2017-01-01

Cytochrome c oxidase (CcO) is a transmembrane protein that uses the free energy of O2 reduction to generate the proton concentration gradient across the membrane. The regulation of competitive proton transfer pathways has been established to be essential to the vectorial transport efficiency of CcO, yet the underlying mechanism at the molecular level remains lacking. Recent studies have highlighted the potential importance of hydration-level change in an internal cavity that connects the proton entrance channel, the site of O2 reduction, and the putative proton exit route. In this work, we use atomistic molecular dynamics simulations to investigate the energetics and timescales associated with the volume fluctuation and hydration-level change in this central cavity. Extensive unrestrained molecular dynamics simulations (accumulatively ∼4 μs) and free energy computations for different chemical states of CcO support a model in which the volume and hydration level of the cavity are regulated by the protonation state of a propionate group of heme a3 and, to a lesser degree, the redox state of heme a and protonation state of Glu286. Markov-state model analysis of ∼2-μs trajectories suggests that hydration-level change occurs on the timescale of 100–200 ns before the proton-loading site is protonated. The computed energetic and kinetic features for the cavity wetting transition suggest that reversible hydration-level change of the cavity can indeed be a key factor that regulates the branching of proton transfer events and therefore contributes to the vectorial efficiency of proton transport. PMID:28973914

Advanced EUV mask and imaging modeling

NASA Astrophysics Data System (ADS)

Evanschitzky, Peter; Erdmann, Andreas

2017-10-01

The exploration and optimization of image formation in partially coherent EUV projection systems with complex source shapes requires flexible, accurate, and efficient simulation models. This paper reviews advanced mask diffraction and imaging models for the highly accurate and fast simulation of EUV lithography systems, addressing important aspects of the current technical developments. The simulation of light diffraction from the mask employs an extended rigorous coupled wave analysis (RCWA) approach, which is optimized for EUV applications. In order to be able to deal with current EUV simulation requirements, several additional models are included in the extended RCWA approach: a field decomposition and a field stitching technique enable the simulation of larger complex structured mask areas. An EUV multilayer defect model including a database approach makes the fast and fully rigorous defect simulation and defect repair simulation possible. A hybrid mask simulation approach combining real and ideal mask parts allows the detailed investigation of the origin of different mask 3-D effects. The image computation is done with a fully vectorial Abbe-based approach. Arbitrary illumination and polarization schemes and adapted rigorous mask simulations guarantee a high accuracy. A fully vectorial sampling-free description of the pupil with Zernikes and Jones pupils and an optimized representation of the diffraction spectrum enable the computation of high-resolution images with high accuracy and short simulation times. A new pellicle model supports the simulation of arbitrary membrane stacks, pellicle distortions, and particles/defects on top of the pellicle. Finally, an extension for highly accurate anamorphic imaging simulations is included. The application of the models is demonstrated by typical use cases.

Membrane Perturbation Induced by Interfacially Adsorbed Peptides

PubMed Central

Zemel, Assaf; Ben-Shaul, Avinoam; May, Sylvio

2004-01-01

The structural and energetic characteristics of the interaction between interfacially adsorbed (partially inserted) α-helical, amphipathic peptides and the lipid bilayer substrate are studied using a molecular level theory of lipid chain packing in membranes. The peptides are modeled as “amphipathic cylinders” characterized by a well-defined polar angle. Assuming two-dimensional nematic order of the adsorbed peptides, the membrane perturbation free energy is evaluated using a cell-like model; the peptide axes are parallel to the membrane plane. The elastic and interfacial contributions to the perturbation free energy of the “peptide-dressed” membrane are evaluated as a function of: the peptide penetration depth into the bilayer's hydrophobic core, the membrane thickness, the polar angle, and the lipid/peptide ratio. The structural properties calculated include the shape and extent of the distorted (stretched and bent) lipid chains surrounding the adsorbed peptide, and their orientational (C-H) bond order parameter profiles. The changes in bond order parameters attendant upon peptide adsorption are in good agreement with magnetic resonance measurements. Also consistent with experiment, our model predicts that peptide adsorption results in membrane thinning. Our calculations reveal pronounced, membrane-mediated, attractive interactions between the adsorbed peptides, suggesting a possible mechanism for lateral aggregation of membrane-bound peptides. As a special case of interest, we have also investigated completely hydrophobic peptides, for which we find a strong energetic preference for the transmembrane (inserted) orientation over the horizontal (adsorbed) orientation. PMID:15189858

Larval food quantity affects development time, survival and adult biological traits that influence the vectorial capacity of Anopheles darlingi under laboratory conditions.

PubMed

Araújo, Maisa da-Silva; Gil, Luiz Herman S; e-Silva, Alexandre de-Almeida

2012-08-02

The incidence of malaria in the Amazon is seasonal and mosquito vectorial capacity parameters, including abundance and longevity, depend on quantitative and qualitative aspects of the larval diet. Anopheles darlingi is a major malaria vector in the Amazon, representing >95% of total Anopheles population present in the Porto Velho region. Despite its importance in the transmission of the Plasmodium parasite, knowledge of the larval biology and ecology is limited. Studies regarding aspects of adult population ecology are more common than studies on larval ecology. However, in order develop effective control strategies and laboratory breeding conditions for this species, more data on the factors affecting vector biology is needed. The aim of the present study is to assess the effects of larval food quantity on the vectorial capacity of An. darling under laboratory conditions. Anopheles darlingi was maintained at 28°C, 80% humidity and exposed to a daily photoperiod of 12 h. Larvae were divided into three experimental groups that were fed either a low, medium, or high food supply (based on the food amounts consumed by other species of culicids). Each experiment was replicated for six times. A cohort of adults were also exposed to each type of diet and assessed for several biological characteristics (e.g. longevity, bite frequency and survivorship), which were used to estimate the vectorial capacity of each experimental group. The group supplied with higher food amounts observed a reduction in development time while larval survival increased. In addition to enhanced longevity, increasing larval food quantity was positively correlated with increasing frequency of bites, longer blood meal duration and wing length, resulting in greater vectorial capacity. However, females had greater longevity than males despite having smaller wings. Overall, several larval and adult biological traits were significantly affected by larval food availability. Greater larval food supply led to enhance larval and production and larger mosquitoes with longer longevity and higher biting frequency. Thus, larval food availability can alter important biological traits that influence the vectorial capacity of An. darlingi.

P-type ATPases as drug targets: tools for medicine and science.

PubMed

Yatime, Laure; Buch-Pedersen, Morten J; Musgaard, Maria; Morth, J Preben; Lund Winther, Anne-Marie; Pedersen, Bjørn P; Olesen, Claus; Andersen, Jens Peter; Vilsen, Bente; Schiøtt, Birgit; Palmgren, Michael G; Møller, Jesper V; Nissen, Poul; Fedosova, Natalya

2009-04-01

P-type ATPases catalyze the selective active transport of ions like H+, Na+, K+, Ca2+, Zn2+, and Cu2+ across diverse biological membrane systems. Many members of the P-type ATPase protein family, such as the Na+,K+-, H+,K+-, Ca2+-, and H+-ATPases, are involved in the development of pathophysiological conditions or provide critical function to pathogens. Therefore, they seem to be promising targets for future drugs and novel antifungal agents and herbicides. Here, we review the current knowledge about P-type ATPase inhibitors and their present use as tools in science, medicine, and biotechnology. Recent structural information on a variety of P-type ATPase family members signifies that all P-type ATPases can be expected to share a similar basic structure and a similar basic machinery of ion transport. The ion transport pathway crossing the membrane lipid bilayer is constructed of two access channels leading from either side of the membrane to the ion binding sites at a central cavity. The selective opening and closure of the access channels allows vectorial access/release of ions from the binding sites. Recent structural information along with new homology modeling of diverse P-type ATPases in complex with known ligands demonstrate that the most proficient way for the development of efficient and selective drugs is to target their ion transport pathway.

Estimation of vectorial capacity of Anopheles minimus Theobald & An. fluviatilis James (Diptera: Culicidae) in a malaria endemic area of Odisha State, India.

PubMed

Gunasekaran, K; Sahu, S S; Jambulingam, P

2014-11-01

Anopheles minimus and An. fluviatilis were incriminated as the major malaria vectors in Keonjhar district of Odisha State recently. This study was carried out to elucidate the potential role of these two vector species in transmission of malaria during different seasons, and vectorial capacity of these species was also estimated. Three hilly and forested villages of Keonjhar district were randomly selected. Vectorial capacity (C) was calculated using the Macdonald's formula as modified by Garret-Jones. The human landing density of the vector species was obtained from all night human landing collections (bait protected by bed-net). Man feeding habit was estimated by multiplying the human blood index with feeding frequency, which was obtained on daily basis from the duration of gonotrophic cycle. The probability of survival through the extrinsic incubation cycle was calculated from the probability of survival through one day and duration of sporogonic cycle. The estimated vectorial capacity of An. minimus varied between 0.014 and 1.09 for Plasmodium falciparum (Pf) and between 0.1 and 1.46 for P. vivax (Pv). The C of An. minimus for both Pf and Pv was higher during rainy season than the other two seasons. The estimated C of An. fluviatilis varied between 0.04 and 1.28 for Pf and between 0.20 and 1.54 for Pv. Based on the estimated values of vectorial capacity of the two vector species, the area could be stratified and such stratification would reflect the difference in the intensity of transmission between different strata and accordingly the appropriate control strategy could be adopted for each stratum.

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

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

Wang Zhen; Bai Jing; Xu Yuhong

2008-07-11

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{sub 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 Mmore » intermediate kinetics, especially the slow component in M intermediate decay. The half-life M{sub 412s} increased significantly in BRs in liposomes containing cationic lipids, while decreased in those in anionic liposomes.« less

Mapping the local organization of cell membranes using excitation-polarization-resolved confocal fluorescence microscopy.

PubMed

Kress, Alla; Wang, Xiao; Ranchon, Hubert; Savatier, Julien; Rigneault, Hervé; Ferrand, Patrick; Brasselet, Sophie

2013-07-02

Fluorescence anisotropy and linear dichroism imaging have been widely used for imaging biomolecular orientational distributions in protein aggregates, fibrillar structures of cells, and cell membranes. However, these techniques do not give access to complete orientational order information in a whole image, because their use is limited to parts of the sample where the average orientation of molecules is known a priori. Fluorescence anisotropy is also highly sensitive to depolarization mechanisms such as those induced by fluorescence energy transfer. A fully excitation-polarization-resolved fluorescence microscopy imaging that relies on the use of a tunable incident polarization and a nonpolarized detection is able to circumvent these limitations. We have developed such a technique in confocal epifluorescence microscopy, giving access to new regions of study in the complex and heterogeneous molecular organization of cell membranes. Using this technique, we demonstrate morphological changes at the subdiffraction scale in labeled COS-7 cell membranes whose cytoskeleton is perturbed. Molecular orientational order is also seen to be affected by cholesterol depletion, reflecting the strong interplay between lipid-packing regions and their nearby cytoskeleton. This noninvasive optical technique can reveal local organization in cell membranes when used as a complement to existing methods such as generalized polarization. Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Orientation selectivity of synaptic input to neurons in mouse and cat primary visual cortex.

PubMed

Tan, Andrew Y Y; Brown, Brandon D; Scholl, Benjamin; Mohanty, Deepankar; Priebe, Nicholas J

2011-08-24

Primary visual cortex (V1) is the site at which orientation selectivity emerges in mammals: visual thalamus afferents to V1 respond equally to all stimulus orientations, whereas their target V1 neurons respond selectively to stimulus orientation. The emergence of orientation selectivity in V1 has long served as a model for investigating cortical computation. Recent evidence for orientation selectivity in mouse V1 opens cortical computation to dissection by genetic and imaging tools, but also raises two essential questions: (1) How does orientation selectivity in mouse V1 neurons compare with that in previously described species? (2) What is the synaptic basis for orientation selectivity in mouse V1? A comparison of orientation selectivity in mouse and in cat, where such measures have traditionally been made, reveals that orientation selectivity in mouse V1 is weaker than in cat V1, but that spike threshold plays a similar role in narrowing selectivity between membrane potential and spike rate. To uncover the synaptic basis for orientation selectivity, we made whole-cell recordings in vivo from mouse V1 neurons, comparing neuronal input selectivity-based on membrane potential, synaptic excitation, and synaptic inhibition-to output selectivity based on spiking. We found that a neuron's excitatory and inhibitory inputs are selective for the same stimulus orientations as is its membrane potential response, and that inhibitory selectivity is not broader than excitatory selectivity. Inhibition has different dynamics than excitation, adapting more rapidly. In neurons with temporally modulated responses, the timing of excitation and inhibition was different in mice and cats.

Orientation Selectivity of Synaptic Input to Neurons in Mouse and Cat Primary Visual Cortex

PubMed Central

Tan (陈勇毅), Andrew Y. Y.; Brown, Brandon D.; Scholl, Benjamin; Mohanty, Deepankar; Priebe, Nicholas J.

2011-01-01

Primary visual cortex (V1) is the site at which orientation selectivity emerges in mammals: visual thalamus afferents to V1 respond equally to all stimulus orientations whereas their target V1 neurons respond selectively to stimulus orientation. The emergence of orientation selectivity in V1 has long served as a model for investigating cortical computation. Recent evidence for orientation selectivity in mouse V1 opens cortical computation to dissection by genetic and imaging tools, but also raises two essential questions: 1) how does orientation selectivity in mouse V1 neurons compare with that in previously described species? 2) what is the synaptic basis for orientation selectivity in mouse V1? A comparison of orientation selectivity in mouse and in cat, where such measures have traditionally been made, reveals that orientation selectivity in mouse V1 is weaker than in cat V1, but that spike threshold plays a similar role in narrowing selectivity between membrane potential and spike rate. To uncover the synaptic basis for orientation selectivity, we made whole-cell recordings in vivo from mouse V1 neurons, comparing neuronal input selectivity - based on membrane potential, synaptic excitation, and synaptic inhibition - to output selectivity based on spiking. We found that a neuron's excitatory and inhibitory inputs are selective for the same stimulus orientations as is its membrane potential response, and that inhibitory selectivity is not broader than excitatory selectivity. Inhibition has different dynamics than excitation, adapting more rapidly. In neurons with temporally modulated responses, the timing of excitation and inhibition was different in mice and cats. PMID:21865476

Synthesis of focused beam with controllable arbitrary homogeneous polarization using engineered vectorial optical fields.

PubMed

Rui, Guanghao; Chen, Jian; Wang, Xiaoyan; Gu, Bing; Cui, Yiping; Zhan, Qiwen

2016-10-17

The propagation and focusing properties of light beams continue to remain a research interest owning to their promising applications in physics, chemistry and biological sciences. One of the main challenges to these applications is the control of polarization distribution within the focal volume. In this work, we propose and experimentally demonstrate a method for generating a focused beam with arbitrary homogeneous polarization at any transverse plane. The required input field at the pupil plane of a high numerical aperture objective lens can be found analytically by solving an inverse problem with the Richard-Wolf vectorial diffraction method, and can be experimentally created with a vectorial optical field generator. Focused fields with various polarizations are successfully generated and verified using a Stokes parameter measurement to demonstrate the capability and versatility of proposed technique.

Mosquito genomics. Highly evolvable malaria vectors: the genomes of 16 Anopheles mosquitoes.

PubMed

Neafsey, Daniel E; Waterhouse, Robert M; Abai, Mohammad R; Aganezov, Sergey S; Alekseyev, Max A; Allen, James E; Amon, James; Arcà, Bruno; Arensburger, Peter; Artemov, Gleb; Assour, Lauren A; Basseri, Hamidreza; Berlin, Aaron; Birren, Bruce W; Blandin, Stephanie A; Brockman, Andrew I; Burkot, Thomas R; Burt, Austin; Chan, Clara S; Chauve, Cedric; Chiu, Joanna C; Christensen, Mikkel; Costantini, Carlo; Davidson, Victoria L M; Deligianni, Elena; Dottorini, Tania; Dritsou, Vicky; Gabriel, Stacey B; Guelbeogo, Wamdaogo M; Hall, Andrew B; Han, Mira V; Hlaing, Thaung; Hughes, Daniel S T; Jenkins, Adam M; Jiang, Xiaofang; Jungreis, Irwin; Kakani, Evdoxia G; Kamali, Maryam; Kemppainen, Petri; Kennedy, Ryan C; Kirmitzoglou, Ioannis K; Koekemoer, Lizette L; Laban, Njoroge; Langridge, Nicholas; Lawniczak, Mara K N; Lirakis, Manolis; Lobo, Neil F; Lowy, Ernesto; MacCallum, Robert M; Mao, Chunhong; Maslen, Gareth; Mbogo, Charles; McCarthy, Jenny; Michel, Kristin; Mitchell, Sara N; Moore, Wendy; Murphy, Katherine A; Naumenko, Anastasia N; Nolan, Tony; Novoa, Eva M; O'Loughlin, Samantha; Oringanje, Chioma; Oshaghi, Mohammad A; Pakpour, Nazzy; Papathanos, Philippos A; Peery, Ashley N; Povelones, Michael; Prakash, Anil; Price, David P; Rajaraman, Ashok; Reimer, Lisa J; Rinker, David C; Rokas, Antonis; Russell, Tanya L; Sagnon, N'Fale; Sharakhova, Maria V; Shea, Terrance; Simão, Felipe A; Simard, Frederic; Slotman, Michel A; Somboon, Pradya; Stegniy, Vladimir; Struchiner, Claudio J; Thomas, Gregg W C; Tojo, Marta; Topalis, Pantelis; Tubio, José M C; Unger, Maria F; Vontas, John; Walton, Catherine; Wilding, Craig S; Willis, Judith H; Wu, Yi-Chieh; Yan, Guiyun; Zdobnov, Evgeny M; Zhou, Xiaofan; Catteruccia, Flaminia; Christophides, George K; Collins, Frank H; Cornman, Robert S; Crisanti, Andrea; Donnelly, Martin J; Emrich, Scott J; Fontaine, Michael C; Gelbart, William; Hahn, Matthew W; Hansen, Immo A; Howell, Paul I; Kafatos, Fotis C; Kellis, Manolis; Lawson, Daniel; Louis, Christos; Luckhart, Shirley; Muskavitch, Marc A T; Ribeiro, José M; Riehle, Michael A; Sharakhov, Igor V; Tu, Zhijian; Zwiebel, Laurence J; Besansky, Nora J

2015-01-02

Variation in vectorial capacity for human malaria among Anopheles mosquito species is determined by many factors, including behavior, immunity, and life history. To investigate the genomic basis of vectorial capacity and explore new avenues for vector control, we sequenced the genomes of 16 anopheline mosquito species from diverse locations spanning ~100 million years of evolution. Comparative analyses show faster rates of gene gain and loss, elevated gene shuffling on the X chromosome, and more intron losses, relative to Drosophila. Some determinants of vectorial capacity, such as chemosensory genes, do not show elevated turnover but instead diversify through protein-sequence changes. This dynamism of anopheline genes and genomes may contribute to their flexible capacity to take advantage of new ecological niches, including adapting to humans as primary hosts. Copyright © 2015, American Association for the Advancement of Science.

Dual orientation of the outer membrane lipoprotein P6 of nontypeable haemophilus influenzae.

PubMed

Michel, Lea Vacca; Snyder, Joy; Schmidt, Rachel; Milillo, Jennifer; Grimaldi, Kyle; Kalmeta, Breanna; Khan, M Nadeem; Sharma, Sharad; Wright, Leslie Kate; Pichichero, Michael E

2013-07-01

The majority of outer membrane (OM) lipoproteins in Gram-negative bacteria are tethered to the membrane via an attached lipid moiety and oriented facing in toward the periplasmic space; a few lipoproteins have been shown to be surface exposed. The outer membrane lipoprotein P6 from the Gram-negative pathogenic bacterium nontypeable Haemophilus influenzae (NTHi) is surface exposed and a leading vaccine candidate for prevention of NTHi infections. However, we recently found that P6 is not a transmembrane protein as previously thought (L. V. Michel, B. Kalmeta, M. McCreary, J. Snyder, P. Craig, M. E. Pichichero, Vaccine 29:1624-1627, 2011). Here we pursued studies to show that P6 has a dual orientation, existing infrequently as surface exposed and predominantly as internally oriented toward the periplasmic space. Flow cytometry using three monoclonal antibodies with specificity for P6 showed surface staining of whole NTHi cells. Confocal microscopy imaging confirmed that antibodies targeted surface-exposed P6 of intact NTHi cells and not internal P6 in membrane-compromised or dead cells. Western blots of two wild-type NTHi strains and a mutant NTHi strain that does not express P6 showed that P6 antibodies do not detect a promiscuous epitope on NTHi. Depletion of targets to nonlipidated P6 significantly decreased bactericidal activity of human serum. Protease digestion of surface-exposed P6 demonstrated that P6 is predominantly internally localized in a manner similar to its homologue Pal in Escherichia coli. We conclude that P6 of NTHi is likely inserted into the OM in two distinct orientations, with the predominant orientation facing in toward the periplasm.

Filter performance parameters for vectorial high-aperture wave fields.

PubMed

Sheppard, Colin J R; Martinez-Corral, M

2008-03-01

Performance parameters have been presented that can be used to compare the focusing performance of different optical systems, including the effect of pupil filters. These were originally given for the paraxial case and recently extended to the high-aperture scalar regime. We generalize these parameters to the full vectorial case for an aplanatic optical system illuminated by a plane-polarized wave. The behavior of different optical systems is compared.

Correlation functions of main-chain polymer nematics constrained by tensorial and vectorial conservation laws

NASA Astrophysics Data System (ADS)

Svenšek, Daniel; Podgornik, Rudolf

2015-09-01

We present and analyze correlation functions of a main-chain polymer nematic in a continuum worm-like chain description for two types of constraints formalized by the tensorial and vectorial conservation laws, both originating in the microscopic chain integrity, i.e., the connectivity of the polymer chains. In particular, our aim is to identify the features of the correlation functions that are most susceptible to the differences between the two constraints. Besides the density and director autocorrelations in both the tensorial and vectorial cases, we calculate also the density-director correlation functions, the latter being a direct signature of the presence of a specific constraint. Its amplitude is connected to the strength of the constraint and is zero if none of the constraints are present, i.e., for a standard non-polymeric nematic. Generally, the correlation functions with the constraints differ substantially from the correlation functions in the non-polymeric case, if the constraints are strong which in practice requires long chains. Moreover, for the tensorial conservation law to be well distinguishable from the vectorial one, the chain persistence length should be much smaller than the total length of the chain, so that hairpins (chain backfolding) are numerous and the polar order is small.

Quantitative Vectorial Magnetic Imaging of Multi Domain Rock Forming Minerals using Nitrogen-Vacancy Centers in Diamond

NASA Astrophysics Data System (ADS)

Shaar, R.; Farchi, E.; Farfurnik, D.; Ebert, Y.; Haim, G.; Bar-Gill, N.

2017-12-01

Magnetization in rock samples is crucial for paleomagnetometry research, as it harbors valuable geological information on long term processes, such as tectonic movements and the formation of oceans and continents. Nevertheless, current techniques are limited in their ability to measure high spatial resolution and high-sensitivity quantitative vectorial magnetic signatures from individual minerals and micrometer scale samples. As a result, our understanding of bulk rock magnetization is limited, specifically for the case of multi-domain minerals. In this work we use a newly developed nitrogen-vacancy magnetic microscope, capable of quantitative vectorial magnetic imaging with optical resolution. We demonstrate direct imaging of the vectorial magnetic field of a single, multi-domain dendritic magnetite, as well as the measurement and calculation of the weak magnetic moments of an individual grain on the micron scale. Our results were measured in a standoff distance of 3-10 μm, with 350 nm spatial resolution, magnetic sensitivity of 6 μT/√(Hz) and a field of view of 35 μm. The results presented here show the capabilities and the future potential of NV microscopy in measuring the magnetic signals of individual micrometer scale grains. These outcomes pave the way for future applications in paleomagnetometry, and for the fundamental understanding of magnetization in multi-domain samples.

[3D modeling of the female pelvis by Computer-Assisted Anatomical Dissection: Applications and perspectives].

PubMed

Balaya, V; Uhl, J-F; Lanore, A; Salachas, C; Samoyeau, T; Ngo, C; Bensaid, C; Cornou, C; Rossi, L; Douard, R; Bats, A-S; Lecuru, F; Delmas, V

2016-05-01

To achieve a 3D vectorial model of a female pelvis by Computer-Assisted Anatomical Dissection and to assess educationnal and surgical applications. From the database of "visible female" of Visible Human Project(®) (VHP) of the "national library of medicine" NLM (United States), we used 739 transverse anatomical slices of 0.33mm thickness going from L4 to the trochanters. The manual segmentation of each anatomical structures was done with Winsurf(®) software version 4.3. Each anatomical element was built as a separate vectorial object. The whole colored-rendered vectorial model with realistic textures was exported in 3Dpdf format to allow a real time interactive manipulation with Acrobat(®) pro version 11 software. Each element can be handled separately at any transparency, which allows an anatomical learning by systems: skeleton, pelvic organs, urogenital system, arterial and venous vascularization. This 3D anatomical model can be used as data bank to teach of the fundamental anatomy. This 3D vectorial model, realistic and interactive constitutes an efficient educational tool for the teaching of the anatomy of the pelvis. 3D printing of the pelvis is possible with the new printers. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

[Vectorial and congenital transmission of Trypanosoma cruzi in Las Lomitas, Formosa].

PubMed

Sosa-Estani, Sergio; Dri, Lucía; Touris, Cecilia; Abalde, Sergio; Dell'arciprete, Ana; Braunstein, Jose

2009-01-01

Chagas disease, caused by Trypanosoma cruzi, is a major cause of morbidity and mortality in Latin America. The objective of this study was to describe the rate of infestation in four aboriginal communities in Las Lomitas (Great Chaco Region), Formosa, Argentina; the rate of infection in children residing in these communities, in blood donors and in pregnant women who received care at the Hospital Las Lomitas, as well as the rate of congenital infection in children born to women infected during the study period. The rate of infestation of 172 households evaluated in 2006 reached 32%. Prevalence of infection among 445 people was 17.5% and in children under 5 years old it was 8.6%. The rate of infection reached 18.6% in blood donors and 29.1% in pregnant women. The rate of infection among 47 children born to infected women, and living in residences under vectorial surveillance was 17.0%. These infections were considered as congenital. This study showed indexes compatible with active vectorial transmission at the beginning. After vectorial control with insecticides the infestation rate has been reduced to 3.3%. The local health system has introduced high impact procedures of primary and secondary prevention in order to prevent new cases and to treat infected people.

Boron removal in radioactive liquid waste by forward osmosis membrane

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

Doo Seong Hwang; Hei Min Choi; Kune Woo Lee

2013-07-01

This study investigated the treatment of boric acid contained in liquid radioactive waste using a forward osmosis membrane. The boron permeation through the membrane depends on the type of membrane, membrane orientation, pH of the feed solution, salt and boron concentration in the feed solution, and osmotic pressure of the draw solution. The boron flux begins to decline from pH 7 and increases with an increase of the osmotic driving force. The boron flux decreases slightly with the salt concentration, but is not heavily influenced by a low salt concentration. The boron flux increases linearly with the concentration of boron.more » No element except for boron was permeated through the FO membrane in the multi-component system. The maximum boron flux is obtained in an active layer facing a draw solution orientation of the CTA-ES membrane under conditions of less than pH 7 and high osmotic pressure. (authors)« less

The Molecular Structure of Human Red Blood Cell Membranes from Highly Oriented, Solid Supported Multi-Lamellar Membranes

PubMed Central

Himbert, Sebastian; Alsop, Richard J.; Rose, Markus; Hertz, Laura; Dhaliwal, Alexander; Moran-Mirabal, Jose M.; Verschoor, Chris P.; Bowdish, Dawn M. E.; Kaestner, Lars; Wagner, Christian; Rheinstädter, Maikel C.

2017-01-01

We prepared highly oriented, multi-lamellar stacks of human red blood cell (RBC) membranes applied on silicon wafers. RBC ghosts were prepared by hemolysis and applied onto functionalized silicon chips and annealed into multi-lamellar RBC membranes. High resolution X-ray diffraction was used to determine the molecular structure of the stacked membranes. We present direct experimental evidence that these RBC membranes consist of nanometer sized domains of integral coiled-coil peptides, as well as liquid ordered (lo) and liquid disordered (ld) lipids. Lamellar spacings, membrane and hydration water layer thicknesses, areas per lipid tail and domain sizes were determined. The common drug aspirin was added to the RBC membranes and found to interact with RBC membranes and preferably partition in the head group region of the lo domain leading to a fluidification of the membranes, i.e., a thinning of the bilayers and an increase in lipid tail spacing. Our results further support current models of RBC membranes as patchy structures and provide unprecedented structural details of the molecular organization in the different domains. PMID:28045119

The Molecular Structure of Human Red Blood Cell Membranes from Highly Oriented, Solid Supported Multi-Lamellar Membranes

NASA Astrophysics Data System (ADS)

Himbert, Sebastian; Alsop, Richard J.; Rose, Markus; Hertz, Laura; Dhaliwal, Alexander; Moran-Mirabal, Jose M.; Verschoor, Chris P.; Bowdish, Dawn M. E.; Kaestner, Lars; Wagner, Christian; Rheinstädter, Maikel C.

2017-01-01

We prepared highly oriented, multi-lamellar stacks of human red blood cell (RBC) membranes applied on silicon wafers. RBC ghosts were prepared by hemolysis and applied onto functionalized silicon chips and annealed into multi-lamellar RBC membranes. High resolution X-ray diffraction was used to determine the molecular structure of the stacked membranes. We present direct experimental evidence that these RBC membranes consist of nanometer sized domains of integral coiled-coil peptides, as well as liquid ordered (lo) and liquid disordered (ld) lipids. Lamellar spacings, membrane and hydration water layer thicknesses, areas per lipid tail and domain sizes were determined. The common drug aspirin was added to the RBC membranes and found to interact with RBC membranes and preferably partition in the head group region of the lo domain leading to a fluidification of the membranes, i.e., a thinning of the bilayers and an increase in lipid tail spacing. Our results further support current models of RBC membranes as patchy structures and provide unprecedented structural details of the molecular organization in the different domains.

Orientational preferences of neighboring helices can drive ER insertion of a marginally hydrophobic transmembrane helix

PubMed Central

Öjemalm, Karin; Halling, Katrin K.; Nilsson, IngMarie; von Heijne, Gunnar

2013-01-01

Summary α-helical integral membrane proteins critically depend on the correct insertion of their transmembrane α-helices into the lipid bilayer for proper folding, yet a surprisingly large fraction of the transmembrane α-helices in multispanning integral membrane proteins are not sufficiently hydrophobic to insert into the target membrane by themselves. How can such marginally hydrophobic segments nevertheless form transmembrane helices in the folded structure? Here, we show that a transmembrane helix with a strong orientational preference (Ncyt-Clum or Nlum-Ccyt) can both increase and decrease the hydrophobicity threshold for membrane insertion of a neighboring, marginally hydrophobic helix. This effect helps explain the ‘missing hydrophobicity’ in polytopic membrane proteins. PMID:22281052

A recommended procedure for the preparation of oriented clay-mineral specimens for X-ray diffraction analysis; modifications to Drever's filter-membrane peel technique

USGS Publications Warehouse

Pollastro, R.M.

1982-01-01

Extremely well-oriented clay mineral mounts for X-ray diffraction analysis can be prepared quickly and without introducing segregation using the filter-membrane peel technique. Mounting problems encountered with smectite-rich samples can be resolved by using minimal sample and partial air-drying of the clay film before transfer to a glass slide. Samples containing small quantities of clay can produce useful oriented specimens if Teflon masks having more restrictive areas are inserted above the membrane filter during clay deposition. War]page and thermal shock of glass slides can be controlled by using a flat, porous, ceramic plate as a holding surface during heat treatments.

Composite metal membrane

DOEpatents

Peachey, Nathaniel M.; Dye, Robert C.; Snow, Ronny C.; Birdsell, Stephan A.

1998-01-01

A composite metal membrane including a first metal layer of Group IVB met or Group VB metals, the first metal layer sandwiched between two layers of an oriented metal of palladium, platinum or alloys thereof is provided together with a process for the recovery of hydrogen from a gaseous mixture including contacting a hydrogen-containing gaseous mixture with a first side of a nonporous composite metal membrane including a first metal of Group IVB metals or Group VB metals, the first metal layer sandwiched between two layers of an oriented metal of palladium, platinum or alloys thereof, and, separating hydrogen from a second side of the nonporous composite metal membrane.

Trypsin digestion for determining orientation of ATPase in Halobacterium saccharovorum membrane vesicles

NASA Technical Reports Server (NTRS)

Kristjansson, H.; Hochstein, L. I.

1986-01-01

Membranes prepared by low pressure disruption of cells exhibited no ATPase activity in the absence of Triton X-100, although 43% of the total menadione reductase activity was detected. Trypsin digestion reduced menadione reductase activity by 45% whereas ATPase activity was not affected. Disruption of the membrane fraction at higher pressure solubilized about 45% of the ATPase activity. The soluble activity was still enhanced by Triton X-100, suggesting that the detergent, besides disrupting membrane vesicles, also activated the ATPase. The discrepancy in localization of menadione reductase and ATPase activities raised questions regarding the reliability of using a single marker enzyme as an indicator of vesicle orientation.

Composite metal membrane

DOEpatents

Peachey, N.M.; Dye, R.C.; Snow, R.C.; Birdsell, S.A.

1998-04-14

A composite metal membrane including a first metal layer of Group IVB met or Group VB metals, the first metal layer sandwiched between two layers of an oriented metal of palladium, platinum or alloys thereof is provided together with a process for the recovery of hydrogen from a gaseous mixture including contacting a hydrogen-containing gaseous mixture with a first side of a nonporous composite metal membrane including a first metal of Group IVB metals or Group VB metals, the first metal layer sandwiched between two layers of an oriented metal of palladium, platinum or alloys thereof, and, separating hydrogen from a second side of the nonporous composite metal membrane.

Seven Transmembrane Receptors as Shapeshifting Proteins: The Impact of Allosteric Modulation and Functional Selectivity on New Drug Discovery

PubMed Central

Miller, Laurence J.

2010-01-01

It is useful to consider seven transmembrane receptors (7TMRs) as disordered proteins able to allosterically respond to a number of binding partners. Considering 7TMRs as allosteric systems, affinity and efficacy can be thought of in terms of energy flow between a modulator, conduit (the receptor protein), and a number of guests. These guests can be other molecules, receptors, membrane-bound proteins, or signaling proteins in the cytosol. These vectorial flows of energy can yield standard canonical guest allostery (allosteric modification of drug effect), effects along the plane of the cell membrane (receptor oligomerization), or effects directed into the cytosol (differential signaling as functional selectivity). This review discusses these apparently diverse pharmacological effects in terms of molecular dynamics and protein ensemble theory, which tends to unify 7TMR behavior toward cells. Special consideration will be given to functional selectivity (biased agonism and biased antagonism) in terms of mechanism of action and potential therapeutic application. The explosion of technology that has enabled observation of diverse 7TMR behavior has also shown how drugs can have multiple (pluridimensional) efficacies and how this can cause paradoxical drug classification and nomenclatures. PMID:20392808

Transmembrane Pores Formed by Human Antimicrobial Peptide LL-37

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

Qian, Shuo

Human LL-37 is a multifunctional cathelicidin peptide that has shown a wide spectrum of antimicrobial activity by permeabilizing microbial membranes similar to other antimicrobial peptides; however, its molecular mechanism has not been clarified. Two independent experiments revealed LL-37 bound to membranes in the {alpha}-helical form with the axis lying in the plane of membrane. This led to the conclusion that membrane permeabilization by LL-37 is a nonpore carpet-like mechanism of action. Here we report the detection of transmembrane pores induced by LL-37. The pore formation coincided with LL-37 helices aligning approximately normal to the plane of the membrane. We observedmore » an unusual phenomenon of LL-37 embedded in stacked membranes, which are commonly used in peptide orientation studies. The membrane-bound LL-37 was found in the normal orientation only when the membrane spacing in the multilayers exceeded its fully hydrated value. This was achieved by swelling the stacked membranes with excessive water to a swollen state. The transmembrane pores were detected and investigated in swollen states by means of oriented circular dichroism, neutron in-plane scattering, and x-ray lamellar diffraction. The results are consistent with the effect of LL-37 on giant unilamellar vesicles. The detected pores had a water channel of radius 2333 {angstrom}. The molecular mechanism of pore formation by LL-37 is consistent with the two-state model exhibited by magainin and other small pore-forming peptides. The discovery that peptide-membrane interactions in swollen states are different from those in less hydrated states may have implications for other large membrane-active peptides and proteins studied in stacked membranes.« less

Toward the Fabrication of Advanced Nanofiltration Membranes by Controlling Morphologies and Mesochannel Orientations of Hexagonal Lyotropic Liquid Crystals.

PubMed

Wang, Guang; Garvey, Christopher J; Zhao, Han; Huang, Kang; Kong, Lingxue

2017-07-21

Water scarcity has been recognized as one of the major threats to human activity, and, therefore, water purification technologies are increasingly drawing attention worldwide. Nanofiltration (NF) membrane technology has been proven to be an efficient and cost-effective way in terms of the size and continuity of the nanostructure. Using a template based on hexagonal lyotropic liquid crystals (LLCs) and partitioning monomer units within this structure for subsequent photo-polymerisation presents a unique path for the fabrication of NF membranes, potentially producing pores of uniform size, ranging from 1 to 5 nm, and large surface areas. The subsequent orientation of this pore network in a direction normal to a flat polymer film that provides ideal transport properties associated with continuous pores running through the membrane has been achieved by the orientation of hexagonal LLCs through various strategies. This review presents the current progresses on the strategies for structure retention from a hexagonal LLCs template and the up-to-date techniques used for the reorientation of mesochanels for continuity through the whole membrane.

Decoding divergent series in nonparaxial optics.

PubMed

Borghi, Riccardo; Gori, Franco; Guattari, Giorgio; Santarsiero, Massimo

2011-03-15

A theoretical analysis aimed at investigating the divergent character of perturbative series involved in the study of free-space nonparaxial propagation of vectorial optical beams is proposed. Our analysis predicts a factorial divergence for such series and provides a theoretical framework within which the results of recently published numerical experiments concerning nonparaxial propagation of vectorial Gaussian beams find a meaningful interpretation in terms of the decoding operated on such series by the Weniger transformation.

Vectorial laws of refraction and reflection using the cross product and dot product.

PubMed

Tkaczyk, Eric R

2012-03-01

We demonstrate that published vectorial laws of reflection and refraction of light based solely on the cross product do not, in general, uniquely determine the direction of the reflected and refracted waves without additional information. This is because the cross product does not have a unique inverse operation, which is explained in this Letter in linear algebra terms. However, a vector is in fact uniquely determined if both the cross product (vector product) and dot product (scalar product) with a known vector are specified, which can be written as a single equation with a left-invertible matrix. It is thus possible to amend the vectorial laws of reflection and refraction to incorporate both the cross and dot products for a complete specification with unique solution. This enables highly efficient, unambiguous computation of reflected and refracted wave vectors from the incident wave and surface normal. © 2012 Optical Society of America

Vectorial model for guided-mode resonance gratings

NASA Astrophysics Data System (ADS)

Fehrembach, A.-L.; Gralak, B.; Sentenac, A.

2018-04-01

We propose a self-consistent vectorial method, based on a Green's function technique, to describe the resonances that appear in guided-mode resonance gratings. The model provides intuitive expressions of the reflectivity and transmittivity matrices of the structure, involving coupling integrals between the modes of a planar reference structure and radiative modes. When one mode is excited, the diffracted field for a suitable polarization can be written as the sum of a resonant and a nonresonant term, thus extending the intuitive approach used to explain the Fano shape of the resonance in scalar configurations. When two modes are excited, we derive a physical analysis in a configuration which requires a vectorial approach. We provide numerical validations of our model. From a technical point of view, we show how the Green's tensor of our planar reference structure can be expressed as two scalar Green's functions, and how to deal with the singularity of the Green's tensor.

Lossy chaotic electromagnetic reverberation chambers: Universal statistical behavior of the vectorial field

NASA Astrophysics Data System (ADS)

Gros, J.-B.; Kuhl, U.; Legrand, O.; Mortessagne, F.

2016-03-01

The effective Hamiltonian formalism is extended to vectorial electromagnetic waves in order to describe statistical properties of the field in reverberation chambers. The latter are commonly used in electromagnetic compatibility tests. As a first step, the distribution of wave intensities in chaotic systems with varying opening in the weak coupling limit for scalar quantum waves is derived by means of random matrix theory. In this limit the only parameters are the modal overlap and the number of open channels. Using the extended effective Hamiltonian, we describe the intensity statistics of the vectorial electromagnetic eigenmodes of lossy reverberation chambers. Finally, the typical quantity of interest in such chambers, namely, the distribution of the electromagnetic response, is discussed. By determining the distribution of the phase rigidity, describing the coupling to the environment, using random matrix numerical data, we find good agreement between the theoretical prediction and numerical calculations of the response.

Dominant negative RPW8.2 fusion proteins reveal the importance of haustorium-oriented protein trafficking for resistance against powdery mildew in Arabidopsis.

PubMed

Zhang, Qiong; Berkey, Robert; Pan, Zhiyong; Wang, Wenming; Zhang, Yi; Ma, Xianfeng; King, Harlan; Xiao, Shunyuan

2015-01-01

Powdery mildew fungi form feeding structures called haustoria inside epidermal cells of host plants to extract photosynthates for their epiphytic growth and reproduction. The haustorium is encased by an interfacial membrane termed the extrahaustorial membrane (EHM). The atypical resistance protein RPW8.2 from Arabidopsis is specifically targeted to the EHM where RPW8.2 activates haustorium-targeted (thus broad-spectrum) resistance against powdery mildew fungi. EHM-specific localization of RPW8.2 suggests the existence of an EHM-oriented protein/membrane trafficking pathway during EHM biogenesis. However, the importance of this specific trafficking pathway for host defense has not been evaluated via a genetic approach without affecting other trafficking pathways. Here, we report that expression of EHM-oriented, nonfunctional RPW8.2 chimeric proteins exerts dominant negative effect over functional RPW8.2 and potentially over other EHM-localized defense proteins, thereby compromising both RPW8.2-mediated and basal resistance to powdery mildew. Thus, our results highlight the importance of the EHM-oriented protein/membrane trafficking pathway for host resistance against haustorium-forming pathogens such as powdery mildew fungi.

Metrics of cellular and vascular infiltration of human acellular dermal matrix in ventral hernia repairs.

PubMed

Campbell, Kristin Turza; Burns, Nadja K; Ensor, Joe; Butler, Charles E

2012-04-01

Human acellular dermal matrix is used for ventral hernia repair, as it resists infection and remodels by means of surrounding tissue. However, the tissue source and impact of basement membrane on cell and vessel infiltration have not been determined. The authors hypothesized that musculofascia would be the primary tissue source of cells and vessels infiltrating into human acellular dermal matrix and that the basement membrane would inhibit infiltration. Fifty-six guinea pigs underwent inlay human acellular dermal matrix ventral hernia repair with the basement membrane oriented toward or away from the peritoneum. At postoperative weeks 1, 2, or 4, repair sites were completely excised. Histologic and immunohistochemical analyses were performed to quantify cell and vessel density within repair-site zones, including interface (lateral, beneath musculofascia) and center (beneath subcutaneous fat) zones. Cell and vessel quantities were compared as functions of zone, basement membrane orientation, and time. Cellular and vascular infiltration increased over time universally. The interface demonstrated greater mean cell density than the center (weeks 1 and 2, p = 0.01 and p < 0.0001, respectively). Cell density was greater with the basement membrane oriented toward the peritoneum at week 4 (p = 0.02). The interface zone had greater mean vessel density than the center zone at week 4 (p < 0.0001). Orienting the basement membrane toward the peritoneum increased vessel density at week 4 (p = 0.0004). Cellular and vascular infiltration into human acellular dermal matrix for ventral hernia repairs was greater from musculofascia than from subcutaneous fat, and the basement membrane inhibited cellular and vascular infiltration. Human acellular dermal matrix should be placed adjacent to the best vascularizing tissue to improve fibrovascular incorporation.

Functionality in Electrospun Nanofibrous Membranes Based on Fiber's Size, Surface Area, and Molecular Orientation

PubMed Central

Matsumoto, Hidetoshi; Tanioka, Akihiko

2011-01-01

Electrospinning is a versatile method for forming continuous thin fibers based on an electrohydrodynamic process. This method has the following advantages: (i) the ability to produce thin fibers with diameters in the micrometer and nanometer ranges; (ii) one-step forming of the two- or three-dimensional nanofiber network assemblies (nanofibrous membranes); and (iii) applicability for a broad spectrum of molecules, such as synthetic and biological polymers and polymerless sol-gel systems. Electrospun nanofibrous membranes have received significant attention in terms of their practical applications. The major advantages of nanofibers or nanofibrous membranes are the functionalities based on their nanoscaled-size, highly specific surface area, and highly molecular orientation. These functionalities of the nanofibrous membranes can be controlled by their fiber diameter, surface chemistry and topology, and internal structure of the nanofibers. This report focuses on our studies and describes fundamental aspects and applications of electrospun nanofibrous membranes. PMID:24957735

Well-ordered large-area arrays of epitaxial ferroelectric (Bi,La)4Ti3O12 nanostructures fabricated by gold nanotube-membrane lithography

NASA Astrophysics Data System (ADS)

Lee, Sung Kyun; Lee, Woo; Alexe, Marin; Nielsch, Kornelius; Hesse, Dietrich; Gösele, Ulrich

2005-04-01

Two-dimensionally well-ordered, large-area arrays of epitaxial, ferroelectric, La-substituted Bi4Ti3O12 (BLT) nanostructures are prepared using gold nanotube membranes as a liftoff mask. Epitaxial nanostructures with a height of about 65nm and a lateral size of about 150nm, with either (001) ("c-axis") orientation, or mixed (118)/(100) ("non-c-axis") orientation, are obtained on (001)- and (011)-oriented SrTiO3 substrates, respectively. The ferroelectric properties are probed by piezoresponse scanning force microscopy. Non-c-axis-oriented BLT nanostructures show an effective piezoresponse coefficient (2dzz) of about 38.0pm /V, whereas c-axis-oriented structures show one of only about 4.9pm/V.

Cytoskeleton-mediated templating of complex cellulose-scaffolded extracellular structure and its association with oikosins in the urochordate Oikopleura.

PubMed

Sagane, Yoshimasa; Hosp, Julia; Zech, Karin; Thompson, Eric M

2011-05-01

Oriented cellulose deposition is critical to plant patterning and models suggest microtubules constrain cellulose synthase movements through the plasma membrane. Though widespread in plants, urochordates are the only animals that synthesize cellulose. We characterized the distinctive cellulose microfibril scaffold of the larvacean house and its interaction with house structural proteins (oikosins). Targeted disruption of cytoskeletal elements, secretory pathways, and plasma membrane organization, suggested a working model for templating extracellular cellulose microfibrils from animal cells that shows both convergence and differences to plant models. Specialized cortical F-actin arrays template microfibril orientation and glycosylphosphatidylinositol-anchored proteins in lipid rafts may act as scaffolding proteins in microfibril elongation. Microtubules deliver and maintain cellulose synthase complexes to specific cell membrane sites rather than orienting their movement through the membrane. Oikosins are incorporated into house compartments directly above their corresponding cellular field of expression and interact with the cellulose scaffold to a variable extent.

A general melt-injection-decomposition route to oriented metal oxide nanowire arrays

NASA Astrophysics Data System (ADS)

Han, Dongqiang; Zhang, Xinwei; Hua, Zhenghe; Yang, Shaoguang

2016-12-01

In this manuscript, a general melt-injection-decomposition (MID) route has been proposed and realized for the fabrication of oriented metal oxide nanowire arrays. Nitrate was used as the starting materials, which was injected into the nanopores of the anodic aluminum oxide (AAO) membrane through the capillarity action in its liquid state. At higher temperature, the nitrate decomposed into corresponding metal oxide within the nanopores of the AAO membrane. Oriented metal oxide nanowire arrays were formed within the AAO membrane as a result of the confinement of the nanopores. Four kinds of metal oxide (CuO, Mn2O3, Co3O4 and Cr2O3) nanowire arrays are presented here as examples fabricated by this newly developed process. X-ray diffraction, scanning electron microscopy and transmission electron microscopy studies showed clear evidence of the formations of the oriented metal oxide nanowire arrays. Formation mechanism of the metal oxide nanowire arrays is discussed based on the Thermogravimetry and Differential Thermal Analysis measurement results.

The Influence of Atmosphere Parameters on the Signal for Remote Sensing Polarimetric Electro-Optical Systems

NASA Astrophysics Data System (ADS)

Budak, Vladimir P.; Korkin, Sergey V.

2009-03-01

The singularity subtraction on the vectorial modification of spherical harmonics method (VMSH) of the solution of the vectorial radiative transfer equation boundary problem is applied to the problem of influence of atmosphere parameters on the polarimetric system signal. We assume in this model different phase matrices (Mie, Rayleigh, and Henyey-Greenstein), reflecting bottom and particle size distribution. The authors describe the main features of the model and some results of its implementation.

FDTD Modeling and Counteraction to Scintillation Effects in the lonosphere

DTIC Science & Technology

2014-04-05

collection of information if it does not display a currently valid OMB control number. PLEASE DO NOT RETURN YOUR FORM TO THE ABOVE ADDRESS. 1. REPORT...for vectorial diffraction calculations and its numerical implementation,” J. Opt. Soc. Am. A, 23 (3), pp. 713-722, 2006. [21] Coe, R. L. and E. J...Seibel, “Improved near-field calculations using vectorial diffraction integrals in the finite-difference time-domain method,” J. Opt. Soc. Am. A, 28

Hybrid reflection type metasurface of nano-antennas designed for optical needle field generation

NASA Astrophysics Data System (ADS)

Wang, Shiyi; Zhan, Qiwen

2015-03-01

We propose a reflection type metal-insulator-metal (MIM) metasurface composed of hybrid optical antennas for comprehensive spatial engineering the properties of optical fields. Its capability is illustrated with an example to create a radially polarized vectorial beam for optical needle field generation. Functioning as local quarter-wave-plates (QWP), the MIM metasurface is designed to convert circularly polarized incident into local linear polarization to create an overall radial polarization with corresponding binary phases and desired normalized amplitude modulation ranged from 0.07 to 1. To obtain enough degrees of freedom, the optical-antenna layer comprises periodic arrangements of double metallic nano-bars with perpendicular placement and single nano-bars respectively for different amplitude modulation requirements. Both of the antennas enable to introduce π/2 retardation while reaching the desired modulation range both for phase and amplitude. Through adjusting the antennas' geometry and array carefully, we shift the gap-surface plasmon resonances facilitated by optical antennas to realize the manipulation of vectorial properties. Designed at 1064 nm wavelength, the particularly generated vectorial light output can be further tightly focused by a high numerical aperture objective to obtain longitudinally polarized flat-top focal field. The so-called optical needle field is a promising candidate for novel applications that transcend disciplinary boundaries. The proposed metasurface establishes a new class of compact optical components based on nano-scale structures, leading to compound functions for vectorial light generation.

Adult vector control, mosquito ecology and malaria transmission

PubMed Central

Brady, Oliver J.; Godfray, H. Charles J.; Tatem, Andrew J.; Gething, Peter W.; Cohen, Justin M.; McKenzie, F. Ellis; Alex Perkins, T.; Reiner, Robert C.; Tusting, Lucy S.; Scott, Thomas W.; Lindsay, Steven W.; Hay, Simon I.; Smith, David L.

2015-01-01

Background Standard advice regarding vector control is to prefer interventions that reduce the lifespan of adult mosquitoes. The basis for this advice is a decades-old sensitivity analysis of ‘vectorial capacity’, a concept relevant for most malaria transmission models and based solely on adult mosquito population dynamics. Recent advances in micro-simulation models offer an opportunity to expand the theory of vectorial capacity to include both adult and juvenile mosquito stages in the model. Methods In this study we revisit arguments about transmission and its sensitivity to mosquito bionomic parameters using an elasticity analysis of developed formulations of vectorial capacity. Results We show that reducing adult survival has effects on both adult and juvenile population size, which are significant for transmission and not accounted for in traditional formulations of vectorial capacity. The elasticity of these effects is dependent on various mosquito population parameters, which we explore. Overall, control is most sensitive to methods that affect adult mosquito mortality rates, followed by blood feeding frequency, human blood feeding habit, and lastly, to adult mosquito population density. Conclusions These results emphasise more strongly than ever the sensitivity of transmission to adult mosquito mortality, but also suggest the high potential of combinations of interventions including larval source management. This must be done with caution, however, as policy requires a more careful consideration of costs, operational difficulties and policy goals in relation to baseline transmission. PMID:25733562

Occurrence and Characteristics of a Rapid Exchange of Phosphate Oxygens Catalyzed by Sarcoplasmic Reticulum Vesicles

DOE R&D Accomplishments Database

Kanazawa, T.; Boyer, P. D.

1972-01-01

Sarcoplasmic reticulum vesicles isolated from skeletal muscle actively take up Ca{sup ++} from the medium in the presence of Mg{sup ++} and ATP. This transport is coupled to ATP hydrolysis catalyzed by membrane-bound Ca{sup++}, Mg{sup ++}-ATPase which is activated by concurrent presence of Ca{sup ++} and Mg{sup ++}. Considerable informations have accumulated that give insight into the ATPase and its coupling to the calcium transport. The hydrolysis of ATP by this enzyme occurs through a phosphorylated intermediate. Formation and decomposition of the intermediate show vectorial requirements for Ca{sup ++} and Mg{sup ++}, suggesting an intimate involvement of the intermediate in the transport process. ATP synthesis from P{sub i} and ADP coupled to outflow of Ca{sup ++} from sarcoplasmic reticulum vesicles has recently been demonstrated. This indicates the reversibility of the entire process of calcium transport in sarcoplasmic reticulum vesicles.

Quantitation of protein orientation in flow-oriented unilamellar liposomes by linear dichroism

NASA Astrophysics Data System (ADS)

Rajendra, Jascindra; Damianoglou, Angeliki; Hicks, Matthew; Booth, Paula; Rodger, P. Mark; Rodger, Alison

2006-07-01

The linear dichroism of the visible wavelength transitions of retinal have been used to analyse linear dichroism spectra to determine the orientation of aromatic and peptide structural motifs of Bacteriorhodopsin incorporated into unilamellar soy bean liposomes. The results are consistent with the available X-ray data. This proves that visible light absorbing chromophores can be used to analyse linear dichroism data to give the orientation of membrane proteins in membrane mimicking environments. The work has been extended by screening a wide range of hydrophobic molecules with high extinction coefficients in transitions above 300 nm to find molecules that could be used as independent probes of liposome orientation for experiments involving proteins incorporated into liposomes. Three probes were found to have potential for future work: bis-(1,3-dibutylbarbituric acid)pentamethine oxonol (DiBAC 4), retinol and rhodamine B. All three can be used to determine the orientation of the porphyrin of cytochrome c, the aromatic residues of gramicidin and the helices of both proteins. The orientation parameter, S, for the liposomes varied from batch to batch of unilamellar liposomes prepared by extruding through a 100 nm membrane. The value and variation in S was 0.030 ± 0.010. Repeat experiments with the same batch of liposomes showed less variation. Film LD data were measured for DiBAC 4 and rhodamine B to determine the polarisations of their long wavelength transitions.

An age-structured extension to the vectorial capacity model.

PubMed

Novoseltsev, Vasiliy N; Michalski, Anatoli I; Novoseltseva, Janna A; Yashin, Anatoliy I; Carey, James R; Ellis, Alicia M

2012-01-01

Vectorial capacity and the basic reproductive number (R(0)) have been instrumental in structuring thinking about vector-borne pathogen transmission and how best to prevent the diseases they cause. One of the more important simplifying assumptions of these models is age-independent vector mortality. A growing body of evidence indicates that insect vectors exhibit age-dependent mortality, which can have strong and varied affects on pathogen transmission dynamics and strategies for disease prevention. Based on survival analysis we derived new equations for vectorial capacity and R(0) that are valid for any pattern of age-dependent (or age-independent) vector mortality and explore the behavior of the models across various mortality patterns. The framework we present (1) lays the groundwork for an extension and refinement of the vectorial capacity paradigm by introducing an age-structured extension to the model, (2) encourages further research on the actuarial dynamics of vectors in particular and the relationship of vector mortality to pathogen transmission in general, and (3) provides a detailed quantitative basis for understanding the relative impact of reductions in vector longevity compared to other vector-borne disease prevention strategies. Accounting for age-dependent vector mortality in estimates of vectorial capacity and R(0) was most important when (1) vector densities are relatively low and the pattern of mortality can determine whether pathogen transmission will persist; i.e., determines whether R(0) is above or below 1, (2) vector population growth rate is relatively low and there are complex interactions between birth and death that differ fundamentally from birth-death relationships with age-independent mortality, and (3) the vector exhibits complex patterns of age-dependent mortality and R(0) ∼ 1. A limiting factor in the construction and evaluation of new age-dependent mortality models is the paucity of data characterizing vector mortality patterns, particularly for free ranging vectors in the field.

An Age-Structured Extension to the Vectorial Capacity Model

PubMed Central

Novoseltsev, Vasiliy N.; Michalski, Anatoli I.; Novoseltseva, Janna A.; Yashin, Anatoliy I.; Carey, James R.; Ellis, Alicia M.

2012-01-01

Background Vectorial capacity and the basic reproductive number (R0) have been instrumental in structuring thinking about vector-borne pathogen transmission and how best to prevent the diseases they cause. One of the more important simplifying assumptions of these models is age-independent vector mortality. A growing body of evidence indicates that insect vectors exhibit age-dependent mortality, which can have strong and varied affects on pathogen transmission dynamics and strategies for disease prevention. Methodology/Principal Findings Based on survival analysis we derived new equations for vectorial capacity and R0 that are valid for any pattern of age-dependent (or age–independent) vector mortality and explore the behavior of the models across various mortality patterns. The framework we present (1) lays the groundwork for an extension and refinement of the vectorial capacity paradigm by introducing an age-structured extension to the model, (2) encourages further research on the actuarial dynamics of vectors in particular and the relationship of vector mortality to pathogen transmission in general, and (3) provides a detailed quantitative basis for understanding the relative impact of reductions in vector longevity compared to other vector-borne disease prevention strategies. Conclusions/Significance Accounting for age-dependent vector mortality in estimates of vectorial capacity and R0 was most important when (1) vector densities are relatively low and the pattern of mortality can determine whether pathogen transmission will persist; i.e., determines whether R0 is above or below 1, (2) vector population growth rate is relatively low and there are complex interactions between birth and death that differ fundamentally from birth-death relationships with age-independent mortality, and (3) the vector exhibits complex patterns of age-dependent mortality and R0∼1. A limiting factor in the construction and evaluation of new age-dependent mortality models is the paucity of data characterizing vector mortality patterns, particularly for free ranging vectors in the field. PMID:22724022

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

PubMed

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

2016-05-01

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

Helix Fraying and Lipid-Dependent Structure of a Short Amphipathic Membrane-Bound Peptide Revealed by Solid-State NMR.

PubMed

Strandberg, Erik; Grau-Campistany, Ariadna; Wadhwani, Parvesh; Bürck, Jochen; Rabanal, Francesc; Ulrich, Anne S

2018-06-14

The amphipathic α-helical peptide KIA14 [(KIAGKIA) 2 -NH 2 ] was studied in membranes using circular dichroism and solid-state NMR spectroscopy to obtain global as well as local structural information. By analyzing 2 H NMR data from 10 analogues of KIA14 that were selectively labeled with Ala- d 3 , those positions that are properly folded into a helix could be determined within the membrane-bound peptide. The N-terminus was found to be unraveled, whereas positions 4-14 formed an ideal helix all the way to the C-terminus. The helicity did not change when Gly residues were replaced by Ala- d 3 but was reduced when Ile was replaced, indicating that large hydrophobic residues are required for membrane binding and helix formation. The reduced helicity was strongly correlated with a decrease in peptide-induced leakage from lipid vesicles. The orientation of the short KIA14 peptide was assessed in several lipid systems and compared with that of the longer KIA21 sequence [(KIAGKIA) 3 -NH 2 ]. In 1,2-dioleoyl- sn-glycero-3-phosphatidylcholine, both peptides are aligned flat on the membrane surface, whereas in 1,2-dimyristoyl- sn-glycero-3-phosphatidylcholine (DMPC)/1-myristoyl-2-hydroxy- sn-glycero-3-phosphatidylcholine (lyso-MPC) both are inserted into the membrane in an upright orientation. These two types of lipid systems had been selected for their strongly negative and positive spontaneous curvature, respectively. We propose that in these cases, the peptide orientation is largely determined by the lipid properties. On the other hand, in plain DMPC and 1,2-dilauroyl- sn-glycero-3-phosphatidylcholine, which have only a slight positive curvature, a marked difference in orientation is evident: the short KIA14 lies almost flat on the membrane surface, whereas the longer KIA21 is more tilted. We thus propose that out of the lipid systems tested here, DMPC (with hardly any curvature) is the least biased lipid system in which peptide orientation and realignment can be studied, allowing to compare and discriminate the intrinsic effects of the properties of the peptides as such.

[Current status of the knowledge on Moroccan anophelines (Diptera: Culicidae): systematic, geographical distribution and vectorial competence].

PubMed

Faraj, C; Ouahabi, S; Adlaoui, E; Elaouad, R

2010-10-01

This bibliographical study, based on published works, ministry of Health Reports, exploitation of the database relative to the entomological surveillance conducted in the framework of the National Malaria Control Program, as well as unpublished results obtained within the framework of the European project "Emerging disease in a changing European environment", summarizes and completes with new data current knowledge on the systematics, the distribution and the vectorial competence of moroccan anophelines. Copyright © 2010 Elsevier Masson SAS. All rights reserved.

Random vectors and spatial analysis by geostatistics for geotechnical applications

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

Young, D.S.

1987-08-01

Geostatistics is extended to the spatial analysis of vector variables by defining the estimation variance and vector variogram in terms of the magnitude of difference vectors. Many random variables in geotechnology are in vectorial terms rather than scalars, and its structural analysis requires those sample variable interpolations to construct and characterize structural models. A better local estimator will result in greater quality of input models; geostatistics can provide such estimators; kriging estimators. The efficiency of geostatistics for vector variables is demonstrated in a case study of rock joint orientations in geological formations. The positive cross-validation encourages application of geostatistics tomore » spatial analysis of random vectors in geoscience as well as various geotechnical fields including optimum site characterization, rock mechanics for mining and civil structures, cavability analysis of block cavings, petroleum engineering, and hydrologic and hydraulic modelings.« less

Effects of spacer orientations on the cake formation during membrane fouling: Quantitative analysis based on 3D OCT imaging.

PubMed

Liu, Xin; Li, Weiyi; Chong, Tzyy Haur; Fane, Anthony G

2017-03-01

Spacer design plays an important role in improving the performance of membrane processes for water/wastewater treatment. This work focused on a fundamental issue of spacer design, i.e., investigating the effects of spacer orientations on the fouling behavior during a membrane process. A series of fouling experiments with different spacer orientation were carried out to in situ characterize the formation of a cake layer in a spacer unit cell via 3D optical coherence tomography (OCT) imaging. The cake layers formed at different times were digitalized for quantitatively analyzing the variation in the cake morphology as a function of time. In particular, the local deposition rates were evaluated to determine the active regions where the instantaneous changes in deposit thickness were significant. The characterization results indicate that varying the spacer orientation could substantially change the evolution of membrane fouling by particulate foulants and thereby result in a cake layer with various morphologies; the competition between growth and erosion at different locations would instantaneously respond to the micro-hydrodynamic environment that might change with time. This work confirms that the OCT-based characterization method is a powerful tool for exploring novel spacer design. Copyright © 2016 Elsevier Ltd. All rights reserved.

The Drive to Life on Wet and Icy Worlds

PubMed Central

Barge, Laura M.; Bhartia, Rohit; Bocanegra, Dylan; Bracher, Paul J.; Branscomb, Elbert; Kidd, Richard; McGlynn, Shawn; Meier, David H.; Nitschke, Wolfgang; Shibuya, Takazo; Vance, Steve; White, Lauren; Kanik, Isik

2014-01-01

Abstract This paper presents a reformulation of the submarine alkaline hydrothermal theory for the emergence of life in response to recent experimental findings. The theory views life, like other self-organizing systems in the Universe, as an inevitable outcome of particular disequilibria. In this case, the disequilibria were two: (1) in redox potential, between hydrogen plus methane with the circuit-completing electron acceptors such as nitrite, nitrate, ferric iron, and carbon dioxide, and (2) in pH gradient between an acidulous external ocean and an alkaline hydrothermal fluid. Both CO2 and CH4 were equally the ultimate sources of organic carbon, and the metal sulfides and oxyhydroxides acted as protoenzymatic catalysts. The realization, now 50 years old, that membrane-spanning gradients, rather than organic intermediates, play a vital role in life's operations calls into question the idea of “prebiotic chemistry.” It informs our own suggestion that experimentation should look to the kind of nanoengines that must have been the precursors to molecular motors—such as pyrophosphate synthetase and the like driven by these gradients—that make life work. It is these putative free energy or disequilibria converters, presumably constructed from minerals comprising the earliest inorganic membranes, that, as obstacles to vectorial ionic flows, present themselves as the candidates for future experiments. Key Words: Methanotrophy—Origin of life. Astrobiology 14, 308–343. The fixation of inorganic carbon into organic material (autotrophy) is a prerequisite for life and sets the starting point of biological evolution. (Fuchs, 2011) Further significant progress with the tightly membrane-bound H+-PPase family should lead to an increased insight into basic requirements for the biological transport of protons through membranes and its coupling to phosphorylation. (Baltscheffsky et al., 1999) PMID:24697642

The drive to life on wet and icy worlds.

PubMed

Russell, Michael J; Barge, Laura M; Bhartia, Rohit; Bocanegra, Dylan; Bracher, Paul J; Branscomb, Elbert; Kidd, Richard; McGlynn, Shawn; Meier, David H; Nitschke, Wolfgang; Shibuya, Takazo; Vance, Steve; White, Lauren; Kanik, Isik

2014-04-01

This paper presents a reformulation of the submarine alkaline hydrothermal theory for the emergence of life in response to recent experimental findings. The theory views life, like other self-organizing systems in the Universe, as an inevitable outcome of particular disequilibria. In this case, the disequilibria were two: (1) in redox potential, between hydrogen plus methane with the circuit-completing electron acceptors such as nitrite, nitrate, ferric iron, and carbon dioxide, and (2) in pH gradient between an acidulous external ocean and an alkaline hydrothermal fluid. Both CO2 and CH4 were equally the ultimate sources of organic carbon, and the metal sulfides and oxyhydroxides acted as protoenzymatic catalysts. The realization, now 50 years old, that membrane-spanning gradients, rather than organic intermediates, play a vital role in life's operations calls into question the idea of "prebiotic chemistry." It informs our own suggestion that experimentation should look to the kind of nanoengines that must have been the precursors to molecular motors-such as pyrophosphate synthetase and the like driven by these gradients-that make life work. It is these putative free energy or disequilibria converters, presumably constructed from minerals comprising the earliest inorganic membranes, that, as obstacles to vectorial ionic flows, present themselves as the candidates for future experiments. Key Words: Methanotrophy-Origin of life. Astrobiology 14, 308-343. The fixation of inorganic carbon into organic material (autotrophy) is a prerequisite for life and sets the starting point of biological evolution. (Fuchs, 2011 ) Further significant progress with the tightly membrane-bound H(+)-PPase family should lead to an increased insight into basic requirements for the biological transport of protons through membranes and its coupling to phosphorylation. (Baltscheffsky et al., 1999 ).

Vectorial diffraction properties of THz vortex Bessel beams.

PubMed

Wu, Zhen; Wang, Xinke; Sun, Wenfeng; Feng, Shengfei; Han, Peng; Ye, Jiasheng; Yu, Yue; Zhang, Yan

2018-01-22

A vortex Bessel beam combines the merits of an optical vortex and a Bessel beam, including a spiral wave front and a non-diffractive feature, which has immense application potentials in optical trapping, optical fabrication, optical communications, and so on. Here, linearly and circularly polarized vortex Bessel beams in the terahertz (THz) frequency range are generated by utilizing a THz quarter wave plate, a spiral phase plate, and Teflon axicons with different opening angles. Taking advantage of a THz focal-plane imaging system, vectorial diffraction properties of the THz vortex Bessel beams are comprehensively characterized and discussed, including the transverse (Ex, Ey) and longitudinal (Ez) polarization components. The experimental phenomena are accurately simulated by adopting the vectorial Rayleigh diffraction integral. By varying the opening angle of the axicon, the characteristic parameters of these THz vortex Bessel beams are exhibited and compared, including the light spot size, the diffraction-free range, and the phase evolution process. This work provides the precise experimental and theoretical bases for the comprehension and application of a THz vortex Bessel beam.

On the Milankovitch orbital elements for perturbed Keplerian motion

NASA Astrophysics Data System (ADS)

Rosengren, Aaron J.; Scheeres, Daniel J.

2014-03-01

We consider sets of natural vectorial orbital elements of the Milankovitch type for perturbed Keplerian motion. These elements are closely related to the two vectorial first integrals of the unperturbed two-body problem; namely, the angular momentum vector and the Laplace-Runge-Lenz vector. After a detailed historical discussion of the origin and development of such elements, nonsingular equations for the time variations of these sets of elements under perturbations are established, both in Lagrangian and Gaussian form. After averaging, a compact, elegant, and symmetrical form of secular Milankovitch-like equations is obtained, which reminds of the structure of canonical systems of equations in Hamiltonian mechanics. As an application of this vectorial formulation, we analyze the motion of an object orbiting about a planet (idealized as a point mass moving in a heliocentric elliptical orbit) and subject to solar radiation pressure acceleration (obeying an inverse-square law). We show that the corresponding secular problem is integrable and we give an explicit closed-form solution.

Anopheline Reproductive Biology: Impacts on Vectorial Capacity and Potential Avenues for Malaria Control.

PubMed

Mitchell, Sara N; Catteruccia, Flaminia

2017-12-01

Vectorial capacity is a mathematical approximation of the efficiency of vector-borne disease transmission, measured as the number of new infections disseminated per case per day by an insect vector. Multiple elements of mosquito biology govern their vectorial capacity, including survival, population densities, feeding preferences, and vector competence. Intriguingly, biological pathways essential to mosquito reproductive fitness directly or indirectly influence a number of these elements. Here, we explore this complex interaction, focusing on how the interplay between mating and blood feeding in female Anopheles not only shapes their reproductive success but also influences their ability to sustain Plasmodium parasite development. Central to malaria transmission, mosquito reproductive biology has recently become the focus of research strategies aimed at malaria control, and we discuss promising new methods based on the manipulation of key reproductive steps. In light of widespread resistance to all public health-approved insecticides targeting mosquito reproduction may prove crucial to the success of malaria-eradication campaigns. Copyright © 2017 Cold Spring Harbor Laboratory Press; all rights reserved.

Dynamic Structure of Bombolitin II Bound to Lipid Bilayers as Revealed by Solid-state NMR and Molecular-Dynamics Simulation

PubMed Central

Toraya, Shuichi; Javkhlantugs, Namsrai; Mishima, Daisuke; Nishimura, Katsuyuki; Ueda, Kazuyoshi; Naito, Akira

2010-01-01

Bombolitin II (BLT2) is one of the hemolytic heptadecapeptides originally isolated from the venom of a bumblebee. Structure and orientation of BLT2 bound to 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) membranes were determined by solid-state 31P and 13C NMR spectroscopy. 31P NMR spectra showed that BLT2-DPPC membranes were disrupted into small particles below the gel-to-liquid crystalline phase transition temperature (Tc) and fused to form a magnetically oriented vesicle system where the membrane surface is parallel to the magnetic fields above the Tc. 13C NMR spectra of site-specifically 13C-labeled BLT2 at the carbonyl carbons were observed and the chemical shift anisotropies were analyzed to determine the dynamic structure of BLT2 bound to the magnetically oriented vesicle system. It was revealed that the membrane-bound BLT2 adopted an α-helical structure, rotating around the membrane normal with the tilt angle of the helical axis at 33°. Interatomic distances obtained from rotational-echo double-resonance experiments further showed that BLT2 adopted a straight α-helical structure. Molecular dynamics simulation performed in the BLT2-DPPC membrane system showed that the BLT2 formed a straight α-helix and that the C-terminus was inserted into the membrane. The α-helical axis is tilted 30° to the membrane normal, which is almost the same as the value obtained from solid-state NMR. These results suggest that the membrane disruption induced by BLT2 is attributed to insertion of BLT2 into the lipid bilayers. PMID:21081076

Molecular mechanisms of protein-cholesterol interactions in plasma membranes: Functional distinction between topological (tilted) and consensus (CARC/CRAC) domains.

PubMed

Fantini, Jacques; Di Scala, Coralie; Baier, Carlos J; Barrantes, Francisco J

2016-09-01

The molecular mechanisms that control the multiple possible modes of protein association with membrane cholesterol are remarkably convergent. These mechanisms, which include hydrogen bonding, CH-π stacking and dispersion forces, are used by a wide variety of extracellular proteins (e.g. microbial or amyloid) and membrane receptors. Virus fusion peptides penetrate the membrane of host cells with a tilted orientation that is compatible with a transient interaction with cholesterol; this tilted orientation is also characteristic of the process of insertion of amyloid proteins that subsequently form oligomeric pores in the plasma membrane of brain cells. Membrane receptors that are associated with cholesterol generally display linear consensus binding motifs (CARC and CRAC) characterized by a triad of basic (Lys/Arg), aromatic (Tyr/phe) and aliphatic (Leu/Val) amino acid residues. In some cases, the presence of both CARC and CRAC within the same membrane-spanning domain allows the simultaneous binding of two cholesterol molecules, one in each membrane leaflet. In this review the molecular basis and the functional significance of the different modes of protein-cholesterol interactions in plasma membranes are discussed. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

[Dynamic study of the female levator ani muscle using MRI 3D vectorial modeling].

PubMed

Delmas, Vincent; Ami, Olivier; Iba-Zizen, Marie-Thérèse

2010-06-01

The levator ani muscle has a major role in the female pelvic floor, and is involved in the pathophysiology of pelvic prolapse and stress urinary incontinence. We conducted an anatomical and morphological study of this muscle using dynamic 3D vectorial reconstruction MRI, in order to analyze the contraction of two major components of the levator ani: the iliococcygeus and pubococcygeus. Three volunteer healthy continent nulliparous women aged from 19 to 22 underwent dynamic pelvic MRI. Coronal T2-weighted pelvic images were obtained in the supine position, at rest, holding back, and during Valsalva stress effort. 3D vectorial models were reconstructed by manual segmentation of the source images, and were set up on bony anatomic marks. Iliococcygeus and pubococcygeus volumes were measured in the three positions. Volumetrics, displacement and dynamic morphing changes were analyzed with 3D vectorial animation software. The urogenital hiatus extended more holding back (mean +4.31 mm) than on effort (mean +2.78 mm). The iliococcygeus lowered (mean -3.95 mm) and deviated outward (mean +3.01 mm). The basic tone of the iliococcygeus muscle gives it a dome shape, and its reflex contraction against abdominal strain ensures anal and urinary continence The levator ani is more than a pelvic diaphragm: it is a truly dynamic pelvic floor. Its points of support on the stiff osseous frame allow it to retain the pelvic organs. The levator ani muscle seems to prevent anal prolapse during stress strain.

[Arthropods with vectorial interest in spanish public health].

PubMed

Bueno Marí, Rubén; Moreno Marí, Josefa; Oltra Moscardó, M Teresa; Jiménez Peydró, Ricardo

2009-01-01

Fifteen of the thirty-one Obligatory Communicable Diseases in Spain, exempting those of congenital or neonatal types, can be transmitted by several species of arthropods that are present in our country. Several arthropod orders are the suitable transmitters of tens of bacteria, fungi, virus and protozoa. This fact demands a through of the biology knowledge of these vectors in order to adopt efficient control measures that allow us to reduce the incidence levels of these diseases. Nevertheless, the epidemiological studies shouldn't remain only restricted to the diseases with active transmission cycles in our country. It is necessary to acquire a global vision because of allochton diseases that are perfectly extensible to our territory in the globalization context in which we are situated. All this information is important to know which factors are preventing the disease presence. The aim is to provide the National Epidemiological Surveillance Network with a valuable predictive capacity that allows it to predict the potential arrival of diseases and the consequent strengthening of the spanish Public Health. The goal of this work is to carry out a review of the spanish arthropod fauna with any vectorial interest. The current situation of some of the more important vectorial diseases in our country and the factors related to a resurgence reappearance and/or intensification of those ones are also discussed. Therefore, the study of these inappealable protagonists in our Public Health as an articulatory element in the complex network that any vectorial disease entails is absolutely necessary.

Adult vector control, mosquito ecology and malaria transmission.

PubMed

Brady, Oliver J; Godfray, H Charles J; Tatem, Andrew J; Gething, Peter W; Cohen, Justin M; McKenzie, F Ellis; Alex Perkins, T; Reiner, Robert C; Tusting, Lucy S; Scott, Thomas W; Lindsay, Steven W; Hay, Simon I; Smith, David L

2015-03-01

Standard advice regarding vector control is to prefer interventions that reduce the lifespan of adult mosquitoes. The basis for this advice is a decades-old sensitivity analysis of 'vectorial capacity', a concept relevant for most malaria transmission models and based solely on adult mosquito population dynamics. Recent advances in micro-simulation models offer an opportunity to expand the theory of vectorial capacity to include both adult and juvenile mosquito stages in the model. In this study we revisit arguments about transmission and its sensitivity to mosquito bionomic parameters using an elasticity analysis of developed formulations of vectorial capacity. We show that reducing adult survival has effects on both adult and juvenile population size, which are significant for transmission and not accounted for in traditional formulations of vectorial capacity. The elasticity of these effects is dependent on various mosquito population parameters, which we explore. Overall, control is most sensitive to methods that affect adult mosquito mortality rates, followed by blood feeding frequency, human blood feeding habit, and lastly, to adult mosquito population density. These results emphasise more strongly than ever the sensitivity of transmission to adult mosquito mortality, but also suggest the high potential of combinations of interventions including larval source management. This must be done with caution, however, as policy requires a more careful consideration of costs, operational difficulties and policy goals in relation to baseline transmission. © The Author 2015. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene.

Obstacle-avoiding navigation system

DOEpatents

Borenstein, Johann; Koren, Yoram; Levine, Simon P.

1991-01-01

A system for guiding an autonomous or semi-autonomous vehicle through a field of operation having obstacles thereon to be avoided employs a memory for containing data which defines an array of grid cells which correspond to respective subfields in the field of operation of the vehicle. Each grid cell in the memory contains a value which is indicative of the likelihood, or probability, that an obstacle is present in the respectively associated subfield. The values in the grid cells are incremented individually in response to each scan of the subfields, and precomputation and use of a look-up table avoids complex trigonometric functions. A further array of grid cells is fixed with respect to the vehicle form a conceptual active window which overlies the incremented grid cells. Thus, when the cells in the active window overly grid cell having values which are indicative of the presence of obstacles, the value therein is used as a multiplier of the precomputed vectorial values. The resulting plurality of vectorial values are summed vectorially in one embodiment of the invention to produce a virtual composite repulsive vector which is then summed vectorially with a target-directed vector for producing a resultant vector for guiding the vehicle. In an alternative embodiment, a plurality of vectors surrounding the vehicle are computed, each having a value corresponding to obstacle density. In such an embodiment, target location information is used to select between alternative directions of travel having low associated obstacle densities.

Clustering and negative feedback by endocytosis in planar cell polarity signaling is modulated by ubiquitinylation of prickle.

PubMed

Cho, Bomsoo; Pierre-Louis, Gandhy; Sagner, Andreas; Eaton, Suzanne; Axelrod, Jeffrey D

2015-05-01

The core components of the planar cell polarity (PCP) signaling system, including both transmembrane and peripheral membrane associated proteins, form asymmetric complexes that bridge apical intercellular junctions. While these can assemble in either orientation, coordinated cell polarization requires the enrichment of complexes of a given orientation at specific junctions. This might occur by both positive and negative feedback between oppositely oriented complexes, and requires the peripheral membrane associated PCP components. However, the molecular mechanisms underlying feedback are not understood. We find that the E3 ubiquitin ligase complex Cullin1(Cul1)/SkpA/Supernumerary limbs(Slimb) regulates the stability of one of the peripheral membrane components, Prickle (Pk). Excess Pk disrupts PCP feedback and prevents asymmetry. We show that Pk participates in negative feedback by mediating internalization of PCP complexes containing the transmembrane components Van Gogh (Vang) and Flamingo (Fmi), and that internalization is activated by oppositely oriented complexes within clusters. Pk also participates in positive feedback through an unknown mechanism promoting clustering. Our results therefore identify a molecular mechanism underlying generation of asymmetry in PCP signaling.

Vectorial signalling mechanism required for cell-cell communication during sporulation in Bacillus subtilis.

PubMed

Diez, Veronica; Schujman, Gustavo E; Gueiros-Filho, Frederico J; de Mendoza, Diego

2012-01-01

Spore formation in Bacillus subtilis takes place in a sporangium consisting of two chambers, the forespore and the mother cell, which are linked by pathways of cell-cell communication. One pathway, which couples the proteolytic activation of the mother cell transcription factor σ(E) to the action of a forespore synthesized signal molecule, SpoIIR, has remained enigmatic. Signalling by SpoIIR requires the protein to be exported to the intermembrane space between forespore and mother cell, where it will interact with and activate the integral membrane protease SpoIIGA. Here we show that SpoIIR signal activity as well as the cleavage of its N-terminal extension is strictly dependent on the prespore fatty acid biosynthetic machinery. We also report that a conserved threonine residue (T27) in SpoIIR is required for processing, suggesting that signalling of SpoIIR is dependent on fatty acid synthesis probably because of acylation of T27. In addition, SpoIIR localization in the forespore septal membrane depends on the presence of SpoIIGA. The orchestration of σ(E) activation in the intercellular space by an acylated signal protein provides a new paradigm to ensure local transmission of a weak signal across the bilayer to control cell-cell communication during development. © 2011 Blackwell Publishing Ltd.

Arabidopsis  SABRE and CLASP interact to stabilize cell division plane orientation and planar polarity.

PubMed

Pietra, Stefano; Gustavsson, Anna; Kiefer, Christian; Kalmbach, Lothar; Hörstedt, Per; Ikeda, Yoshihisa; Stepanova, Anna N; Alonso, Jose M; Grebe, Markus

2013-01-01

The orientation of cell division and the coordination of cell polarity within the plane of the tissue layer (planar polarity) contribute to shape diverse multicellular organisms. The root of Arabidopsis thaliana displays regularly oriented cell divisions, cell elongation and planar polarity providing a plant model system to study these processes. Here we report that the SABRE protein, which shares similarity with proteins of unknown function throughout eukaryotes, has important roles in orienting cell division and planar polarity. SABRE localizes at the plasma membrane, endomembranes, mitotic spindle and cell plate. SABRE stabilizes the orientation of CLASP-labelled preprophase band microtubules predicting the cell division plane, and of cortical microtubules driving cell elongation. During planar polarity establishment, sabre is epistatic to clasp at directing polar membrane domains of Rho-of-plant GTPases. Our findings mechanistically link SABRE to CLASP-dependent microtubule organization, shedding new light on the function of SABRE-related proteins in eukaryotes.

Characterization of Interactions between Curcumin and Different Types of Lipid Bilayers by Molecular Dynamics Simulation.

PubMed

Lyu, Yuan; Xiang, Ning; Mondal, Jagannath; Zhu, Xiao; Narsimhan, Ganesan

2018-03-01

Curcumin (CUR) is a natural food ingredient with known ability to target microbial cell membrane. In this study, the interactions of CUR with different types of model lipid bilayers (POPE, POPG, POPC, DOPC, and DPPE), mixtures of model lipid bilayers (POPE/POPG), and biological membrane mimics (Escherichia coli and yeast) were investigated by all-atom explicit solvent molecular dynamics (MD) simulation. CUR readily inserts into different types of model lipid bilayer systems in the liquid crystalline state, staying in the lipid tails region near the interface of lipid head and lipid tail. Parallel orientation to the membrane surface is found to be more probable than perpendicular for CUR, as indicated by the tilt angle distribution. This orientation preference is less significant as the fraction of POPE is increased in the system, likely due to the better water solvation of perpendicular orientation in the POPE bilayer. In E. coli and yeast bilayers, tilt angle distributions were similar to that for POPE/POPG mixed bilayer, with water hydration number around CUR for the former being higher. Insertion of CUR resulted in membrane thinning. The results from these simulations provide insights into the possible differences in membrane disrupting activity of CUR against different types of microorganisms.

Degree of coherence for vectorial electromagnetic fields as the distance between correlation matrices.

PubMed

Luis, Alfredo

2007-04-01

We assess the degree of coherence of vectorial electromagnetic fields in the space-frequency domain as the distance between the cross-spectral density matrix and the identity matrix representing completely incoherent light. This definition is compared with previous approaches. It is shown that this distance provides an upper bound for the degree of coherence and visibility for any pair of scalar waves obtained by linear combinations of the original fields. This same approach emerges when applying a previous definition of global coherence to a Young interferometer.

LOPES-3D - vectorial measurements of radio emission from cosmic ray induced air showers

NASA Astrophysics Data System (ADS)

Huber, D.; Apel, W. D.; Arteaga-Velázquez, J. C.; Bähren, L.; Bekk, K.; Bertaina, M.; Biermann, P. L.; Blümer, J.; Bozdog, H.; Brancus, I. M.; Chiavassa, A.; Daumiller, K.; de Souza, V.; Di Pierro, F.; Doll, P.; Engel, R.; Falcke, H.; Fuchs, B.; Fuhrmann, D.; Gemmeke, H.; Grupen, C.; Haungs, A.; Heck, D.; Hörandel, J. R.; Horneffer, A.; Huege, T.; Isar, P. G.; Kampert, K.-H.; Kang, D.; Krömer, O.; Kuijpers, J.; Link, K.; Łuczak, P.; Ludwig, M.; Mathes, H. J.; Melissas, M.; Morello, C.; Oehlschläger, J.; Palmieri, N.; Pierog, T.; Rautenberg, J.; Rebel, H.; Roth, M.; Rühle, C.; Saftoiu, A.; Schieler, H.; Schmidt, A.; Schröder, F. G.; Sima, O.; Toma, G.; Trinchero, G. C.; Weindl, A.; Wochele, J.; Zabierowski, J.; Zensus, J. A.

2013-05-01

LOPES-3D is able to measure all three components of the electric field vector of the radio emission from air showers. This allows a better comparison with emission models. The measurement of the vertical component increases the sensitivity to inclined showers. By measuring all three components of the electric field vector LOPES-3D demonstrates by how much the reconstruction accuracy of primary cosmic ray parameters increases. Thus LOPES-3D evaluates the usefulness of vectorial measurements for large scale applications.

Use Correlation Coefficients in Gaussian Process to Train Stable ELM Models

DTIC Science & Technology

2015-05-22

confidence interval of prediction y′. There are two parameters that need to be determined in BELM: σ2N and α > 0. BELM effectively controls the over...similarly between h (u) and h (v) with vectorial angle cosine rather than distance between them. The increase of vector dimen- sion will not cause the... vectorial angle cosine approaches 0. Then, we can know that Q I with the increase of L. This reduces the chance of over-fitting. 414 Y. He et al. T a b

Membrane Orientation and Lateral Diffusion of BODIPY-Cholesterol as a Function of Probe Structure

PubMed Central

Solanko, Lukasz M.; Honigmann, Alf; Midtiby, Henrik Skov; Lund, Frederik W.; Brewer, Jonathan R.; Dekaris, Vjekoslav; Bittman, Robert; Eggeling, Christian; Wüstner, Daniel

2013-01-01

Cholesterol tagged with the BODIPY fluorophore via the central difluoroboron moiety of the dye (B-Chol) is a promising probe for studying intracellular cholesterol dynamics. We synthesized a new BODIPY-cholesterol probe (B-P-Chol) with the fluorophore attached via one of its pyrrole rings to carbon-24 of cholesterol (B-P-Chol). Using two-photon fluorescence polarimetry in giant unilamellar vesicles and in the plasma membrane (PM) of living intact and actin-disrupted cells, we show that the BODIPY-groups in B-Chol and B-P-Chol are oriented perpendicular and almost parallel to the bilayer normal, respectively. B-Chol is in all three membrane systems much stronger oriented than B-P-Chol. Interestingly, we found that the lateral diffusion in the PM was two times slower for B-Chol than for B-P-Chol, although we found no difference in lateral diffusion in model membranes. Stimulated emission depletion microscopy, performed for the first time, to our knowledge, with fluorescent sterols, revealed that the difference in lateral diffusion of the BODIPY-cholesterol probes was not caused by anomalous subdiffusion, because diffusion of both analogs in the PM was free but not hindered. Our combined measurements show that the position and orientation of the BODIPY moiety in cholesterol analogs have a severe influence on lateral diffusion specifically in the PM of living cells. PMID:24209853

Evanescent field microscopy techniques for studying dynamics at the surface of living cells

NASA Astrophysics Data System (ADS)

Sund, Susan E.

This thesis presents two distinct optical microscopy techniques for applications in cell biophysics: (a)the extension to living cells of an established technique, total internal reflection/fluorescence recovery after photobleaching (TIR/FRAP) for the first time in imaging mode; and (b)the novel development of polarized total internal reflection fluorescence (p- TIRF) to study membrane orientation in living cells. Although reversible chemistry is crucial to dynamical processes in living cells, relatively little is known about the relevant chemical kinetic rates in vivo. TIR/FRAP, an established technique which can measure reversible biomolecular kinetic rates at surfaces, is extended here to measure kinetic parameters of microinjected rhodamine actin at the cytofacial surface of the plasma membrane of living cultured smooth muscle cells. For the first time, spatial imaging (with a CCD camera) is used in conjunction with TIR/FRAP. TIR/FRAP imaging allows production of spatially resolved images of kinetic data, and calculation of correlation distances, cell-wide gradients, and kinetic parameter dependence on initial fluorescence intensity. In living cells, membrane curvature occurs both in easily imaged large scale morphological features, and also in less visualizable submicroscopic regions of activity such as endocytosis, exocytosis, and cell surface ruffling. A fluorescence microscopic method, p-TIRF, is introduced here to visualize such regions. The method is based on fluorescence of the oriented membrane probe diI- C18-(3) (diI) excited by evanescent field light polarized either perpendicular or parallel to the plane of the substrate coverslip. The excitation efficiency from each polarization depends on the membrane orientation, and thus the ratio of the observed fluorescence excited by these two polarizations vividly shows regions of microscopic and submicroscopic curvature of the membrane. A theoretical background of the technique and experimental verifications are presented in samples of protein solutions, model lipid bilayers, and living cells. Sequential digital images of the polarized TIR fluorescence ratios show spatially-resolved time- course maps of membrane orientations on diI labeled macrophages from which low visibility membrane structures can be identified and quantified. The TIR images are sharpened and contrast-enhanced by deconvoluting them with an experimentally-measured point spread function.

Cell wall biogenesis in Oocystis: experimental alteration of microfibril assembly and orientation.

PubMed

Montezinos, D; Brown, R M

1978-01-01

Cell wall biogenesis in the unicellular green alga Oocystis apiculata has been studied. Under normal growth conditions, a cell wall with ordered microfibrils is synthesized. In each layer there are rows of parallel microfibrils. Layers are nearly perpendicular to each other. Terminal linear synthesizing complexes are located in the plasma membrane, and they are capable of bidirectional synthesis of cellulose microfibrils. Granule bands associated with the inner leaflet of the plasma membrane appear to control the orientation of newly synthesized microfibrils. Subcortical microtubules also are present during wall synthesis. Patterns of cell wall synthesis were studied after treatment with EDTA and EGTA as well as divalent cations (MgSO4, CaSO4, Cacl2). 0.1 M EDTA treatment for 15 min results in the disassociation of the terminal complexes from the ends of microfibrils. EDTA-treated cells followed by 15 min treatment with MgSO4 results in reaggregation of the linear complexes into a paired state, remote from the original ends to which they were associated. After 90 min treatment with MgSO4, normal synthesis resumes. EGTA and calcium salts do not affect the linear complexes or microfibril orientation. Treatments with colchicine and vinblastine sulphate do not depolymerize the microtubles, but the wall microfibril orientation is altered. With colchicine or vinblastine, the change in orientation from layer to layer is inhibited. The process is reversible upon removal of the drugs. Lumicolchicine has no effect upon microfibril orientation, but granule bands are disorganized. Treatment with coumarin, a known inhibitor of cellulose synthesis, causes the loss of visualization of subunits of the terminal complexes. The possibility of the existence of a membrane-associated colchicine-sensitive orientation protein for cellulose microfibrils is discussed. Transmembrane modulation of microfibril synthesis and orientation is presented.

Water Diffusion Mechanism in Carbon Nanotube and Polyamide Nanocomposite Reverse Osmosis Membranes: A Possible Percolation-Hopping Mechanism

NASA Astrophysics Data System (ADS)

Araki, Takumi; Cruz-Silva, Rodolfo; Tejima, Syogo; Ortiz-Medina, Josue; Morelos-Gomez, Aaron; Takeuchi, Kenji; Hayashi, Takuya; Terrones, Mauricio; Endo, Morinobu

2018-02-01

This paper is a contribution to the Physical Review Applied collection in memory of Mildred S. Dresselhaus. The mechanism of water diffusion across reverse osmosis nanocomposite membranes made of carbon nanotubes (CNTs) and aromatic polyamide is not completely understood despite its high potential for desalination applications. While most of the groups have proposed that superflow inside the CNT might positively impact the water flow across membranes, here we show theoretical evidence that this is not likely the case in composite membranes because CNTs are usually oriented parallel to the membrane surface, not to mention that sometimes the nanotube cores are occluded. Instead, we propose an oriented diffusion mechanism that explains the high water permeation by decreasing the diffusion path of water molecules across the membranes, even in the presence of CNTs that behave as impermeable objects. Finally, we provide a comprehensive description of the molecular dynamics occurring in water desalination membranes by considering the bond polarizability caused by dynamic charge transfer and explore the use of molecular-dynamics-derived stochastic diffusion simulations. The proposed water diffusion mechanism offers an alternative and most likely explanation for the high permeation phenomena observed in CNTs and PA nanocomposite membranes, and its understanding can be helpful to design the next generation of reverse osmosis desalination membranes.

Projected impacts of climate change on environmental suitability for malaria transmission in West Africa.

PubMed

Yamana, Teresa K; Eltahir, Elfatih A B

2013-10-01

Climate change is expected to affect the distribution of environmental suitability for malaria transmission by altering temperature and rainfall patterns; however, the local and global impacts of climate change on malaria transmission are uncertain. We assessed the effect of climate change on malaria transmission in West Africa. We coupled a detailed mechanistic hydrology and entomology model with climate projections from general circulation models (GCMs) to predict changes in vectorial capacity, an indication of the risk of human malaria infections, resulting from changes in the availability of mosquito breeding sites and temperature-dependent development rates. Because there is strong disagreement in climate predictions from different GCMs, we focused on the GCM projections that produced the best and worst conditions for malaria transmission in each zone of the study area. Simulation-based estimates suggest that in the desert fringes of the Sahara, vectorial capacity would increase under the worst-case scenario, but not enough to sustain transmission. In the transitional zone of the Sahel, climate change is predicted to decrease vectorial capacity. In the wetter regions to the south, our estimates suggest an increase in vectorial capacity under all scenarios. However, because malaria is already highly endemic among human populations in these regions, we expect that changes in malaria incidence would be small. Our findings highlight the importance of rainfall in shaping the impact of climate change on malaria transmission in future climates. Even under the GCM predictions most conducive to malaria transmission, we do not expect to see a significant increase in malaria prevalence in this region.

Effects of available sugar on the reproductive fitness and vectorial capacity of the malaria vector Anopheles gambiae (Diptera: Culicidae).

PubMed

Gary, R E; Foster, W A

2001-01-01

Although females of most mosquito species are known to use sugar as a necessary source of energy, female Anopheles gambiae Giles sensu stricto are thought to use it facultatively or not at all. However, field evidence of sugar-free living is inconclusive, and the implications for reproductive fitness and vectorial capacity are unknown. To evaluate the role that sugar may play in the ecology of these mosquitoes, mated female An. gambiae in the laboratory were given access to either no food (water only), 10% sucrose, human blood, or human blood + 10% sucrose, and comparisons of daily mortality, fecundity, and biting frequency were made. The effect of sugar availability on vectorial capacity and the intrinsic rate of increase, a measure of fitness, then were determined. Females (pooled and individual) given blood + sugar lived significantly longer than did those on the other diets. Daily fecundity was higher for females given blood alone than for those fed blood + sugar (13 versus 9 eggs per female daily). However, total fecundity and intrinsic rate of increase were not affected by sugar availability. Biting frequency was significantly higher (0.41 versus 0.26 bites per female per day) for females given blood alone. Despite the reduced survivorship, exclusive blood-feeding led to a theoretically higher vectorial capacity for Plasmodium falciparum at 27 degrees C. These data indicate that female An. gambiae could replace sugar with increased blood feeding without suppressing reproductive fitness. Increased blood feeding could, in turn, increase the rate of malaria transmission and may explain the unusual efficiency of this vector.

Multidimensional oriented solid-state NMR experiments enable the sequential assignment of uniformly 15N labeled integral membrane proteins in magnetically aligned lipid bilayers.

PubMed

Mote, Kaustubh R; Gopinath, T; Traaseth, Nathaniel J; Kitchen, Jason; Gor'kov, Peter L; Brey, William W; Veglia, Gianluigi

2011-11-01

Oriented solid-state NMR is the most direct methodology to obtain the orientation of membrane proteins with respect to the lipid bilayer. The method consists of measuring (1)H-(15)N dipolar couplings (DC) and (15)N anisotropic chemical shifts (CSA) for membrane proteins that are uniformly aligned with respect to the membrane bilayer. A significant advantage of this approach is that tilt and azimuthal (rotational) angles of the protein domains can be directly derived from analytical expression of DC and CSA values, or, alternatively, obtained by refining protein structures using these values as harmonic restraints in simulated annealing calculations. The Achilles' heel of this approach is the lack of suitable experiments for sequential assignment of the amide resonances. In this Article, we present a new pulse sequence that integrates proton driven spin diffusion (PDSD) with sensitivity-enhanced PISEMA in a 3D experiment ([(1)H,(15)N]-SE-PISEMA-PDSD). The incorporation of 2D (15)N/(15)N spin diffusion experiments into this new 3D experiment leads to the complete and unambiguous assignment of the (15)N resonances. The feasibility of this approach is demonstrated for the membrane protein sarcolipin reconstituted in magnetically aligned lipid bicelles. Taken with low electric field probe technology, this approach will propel the determination of sequential assignment as well as structure and topology of larger integral membrane proteins in aligned lipid bilayers. © Springer Science+Business Media B.V. 2011

Microbial Pre-exposure and Vectorial Competence of Anopheles Mosquitoes

PubMed Central

Dieme, Constentin; Rotureau, Brice; Mitri, Christian

2017-01-01

Anopheles female mosquitoes can transmit Plasmodium, the malaria parasite. During their aquatic life, wild Anopheles mosquito larvae are exposed to a huge diversity of microbes present in their breeding sites. Later, adult females often take successive blood meals that might also carry different micro-organisms, including parasites, bacteria, and viruses. Therefore, prior to Plasmodium ingestion, the mosquito biology could be modulated at different life stages by a suite of microbes present in larval breeding sites, as well as in the adult environment. In this article, we highlight several naturally relevant scenarios of Anopheles microbial pre-exposure that we assume might impact mosquito vectorial competence for the malaria parasite: (i) larval microbial exposures; (ii) protist co-infections; (iii) virus co-infections; and (iv) pathogenic bacteria co-infections. In addition, significant behavioral changes in African Anopheles vectors have been associated with increasing insecticide resistance. We discuss how these ethological modifications may also increase the repertoire of microbes to which mosquitoes could be exposed, and that might also influence their vectorial competence. Studying Plasmodium–Anopheles interactions in natural microbial environments would efficiently contribute to refining the transmission risks. PMID:29376030

Fully vectorial laser resonator modeling of continuous-wave solid-state lasers including rate equations, thermal lensing and stress-induced birefringence.

PubMed

Asoubar, Daniel; Wyrowski, Frank

2015-07-27

The computer-aided design of high quality mono-mode, continuous-wave solid-state lasers requires fast, flexible and accurate simulation algorithms. Therefore in this work a model for the calculation of the transversal dominant mode structure is introduced. It is based on the generalization of the scalar Fox and Li algorithm to a fully-vectorial light representation. To provide a flexible modeling concept of different resonator geometries containing various optical elements, rigorous and approximative solutions of Maxwell's equations are combined in different subdomains of the resonator. This approach allows the simulation of plenty of different passive intracavity components as well as active media. For the numerically efficient simulation of nonlinear gain, thermal lensing and stress-induced birefringence effects in solid-state active crystals a semi-analytical vectorial beam propagation method is discussed in detail. As a numerical example the beam quality and output power of a flash-lamp-pumped Nd:YAG laser are improved. To that end we compensate the influence of stress-induced birefringence and thermal lensing by an aspherical mirror and a 90° quartz polarization rotator.

A full vectorial generalized discontinuous Galerkin beam propagation method (GDG-BPM) for nonsmooth electromagnetic fields in waveguides

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

Fan Kai; Cai Wei; Ji Xia

2008-07-20

In this paper, we propose a new full vectorial generalized discontinuous Galerkin beam propagation method (GDG-BPM) to accurately handle the discontinuities in electromagnetic fields associated with wave propagations in inhomogeneous optical waveguides. The numerical method is a combination of the traditional beam propagation method (BPM) with a newly developed generalized discontinuous Galerkin (GDG) method [K. Fan, W. Cai, X. Ji, A generalized discontinuous Galerkin method (GDG) for Schroedinger equations with nonsmooth solutions, J. Comput. Phys. 227 (2008) 2387-2410]. The GDG method is based on a reformulation, using distributional variables to account for solution jumps across material interfaces, of Schroedinger equationsmore » resulting from paraxial approximations of vector Helmholtz equations. Four versions of the GDG-BPM are obtained for either the electric or magnetic field components. Modeling of wave propagations in various optical fibers using the full vectorial GDG-BPM is included. Numerical results validate the high order accuracy and the flexibility of the method for various types of interface jump conditions.« less

Direct microscopic observation of forward osmosis membrane fouling.

PubMed

Wang, Yining; Wicaksana, Filicia; Tang, Chuyang Y; Fane, Anthony G

2010-09-15

This study describes the application of a noninvasive direct microscopic observation method for characterizing fouling of a forward osmosis (FO) membrane. The effect of the draw solution concentration, membrane orientation, and feed spacer on FO fouling was systematically investigated in a cross-flow setup using latex particles as model foulant in the feedwater. Higher draw solution (DS) concentrations (and thus increased flux levels) resulted in dramatic increase in the surface coverage by latex particles, suggesting that the critical flux concept might be applicable even for the osmotically driven FO process. Under identical draw solution concentrations, the active-layer-facing-the-feed-solution orientation (AL-FS) experienced significantly less fouling compared to the alternative orientation. This may be explained by the lower water flux in AL-FS, which is consistent with the critical flux concept. The use of a feed spacer not only dramatically enhanced the initial flux of the FO membrane, but also significantly improved the flux stability during FO fouling. Despite such beneficial effects of using the feed spacer, a significant amount of particle accumulation was found near the spacer filament, suggesting further opportunities for improved spacer design. To the best of the authors' knowledge, this is the first direct microscopic observation study on FO fouling.

Organization of fluorescent cholesterol analogs in lipid bilayers - lessons from cyclodextrin extraction.

PubMed

Milles, Sigrid; Meyer, Thomas; Scheidt, Holger A; Schwarzer, Roland; Thomas, Lars; Marek, Magdalena; Szente, Lajos; Bittman, Robert; Herrmann, Andreas; Günther Pomorski, Thomas; Huster, Daniel; Müller, Peter

2013-08-01

To characterize the structure and dynamics of cholesterol in membranes, fluorescent analogs of the native molecule have widely been employed. The cholesterol content in membranes is in general manipulated by using water-soluble cyclodextrins. Since the interactions between cyclodextrins and fluorescent-labeled cholesterol have not been investigated in detail so far, we have compared the cyclodextrin-mediated membrane extraction of three different fluorescent cholesterol analogs (one bearing a NBD and two bearing BODIPY moieties). Extraction of these analogs was followed by measuring the Förster resonance energy transfer between a rhodamine moiety linked to phosphatidylethanolamine and the labeled cholesterol. The extraction kinetics revealed that the analogs are differently extracted from membranes. We examined the orientation of the analogs within the membrane and their influence on lipid condensation using NMR and EPR spectroscopies. Our data indicate that the extraction of fluorescent sterols from membranes is determined by several parameters, including their impact on lipid order, their hydrophobicity, their intermolecular interactions with surrounding lipids, their orientation within the bilayer, and their affinity with the exogenous acceptor. Copyright © 2013 Elsevier B.V. All rights reserved.

Visualization of the membrane engineering concept: evidence for the specific orientation of electroinserted antibodies and selective binding of target analytes.

PubMed

Kokla, Anna; Blouchos, Petros; Livaniou, Evangelia; Zikos, Christos; Kakabakos, Sotiris E; Petrou, Panagiota S; Kintzios, Spyridon

2013-12-01

Membrane engineering is a generic methodology for increasing the selectivity of a cell biosensor against a target molecule, by electroinserting target-specific receptor-like molecules on the cell surface. Previous studies have elucidated the biochemical aspects of the interaction between various analytes (including viruses) and their homologous membrane-engineered cells. In the present study, purified anti-biotin antibodies from a rabbit antiserum along with in-house prepared biotinylated bovine serum albumin (BSA) were used as a model antibody-antigen pair of molecules for facilitating membrane engineering experiments. It was proven, with the aid of fluorescence microscopy, that (i) membrane-engineered cells incorporated the specific antibodies in the correct orientation and that (ii) the inserted antibodies are selectively interacting with the homologous target molecules. This is the first time the actual working concept of membrane engineering has been visualized, thus providing a final proof of the concept behind this innovative process. In addition, the fluorescence microscopy measurements were highly correlated with bioelectric measurements done with the aid of a bioelectric recognition assay. Copyright © 2013 John Wiley & Sons, Ltd.

Modular assembly of synthetic proteins that span the plasma membrane in mammalian cells.

PubMed

Qudrat, Anam; Truong, Kevin

2016-12-09

To achieve synthetic control over how a cell responds to other cells or the extracellular environment, it is important to reliably engineer proteins that can traffic and span the plasma membrane. Using a modular approach to assemble proteins, we identified the minimum necessary components required to engineer such membrane-spanning proteins with predictable orientation in mammalian cells. While a transmembrane domain (TM) fused to the N-terminus of a protein is sufficient to traffic it to the endoplasmic reticulum (ER), an additional signal peptidase cleavage site downstream of this TM enhanced sorting out of the ER. Next, a second TM in the synthetic protein helped anchor and accumulate the membrane-spanning protein on the plasma membrane. The orientation of the components of the synthetic protein were determined through measuring intracellular Ca 2+ signaling using the R-GECO biosensor and through measuring extracellular quenching of yellow fluorescent protein variants by saturating acidic and salt conditions. This work forms the basis of engineering novel proteins that span the plasma membrane to potentially control intracellular responses to extracellular conditions.

Lipid membrane-assisted condensation and assembly of amphiphilic Janus particles

DOE PAGES

Chambers, Mariah; Mallory, Stewart Anthony; Malone, Heather; ...

2016-01-01

Amphiphilic Janus particles self-assemble into complex metastructures, but little is known about how their assembly might be modified by weak interactions with a nearby biological membrane surface. Here, we report an integrated experimental and molecular dynamics simulation study to investigate the self-assembly of amphiphilic Janus particles on a lipid membrane. We created an experimental system in which Janus particles are allowed to self-assemble in the same medium where zwitterionic lipids form giant unilamellar vesicles (GUVs). Janus particles spontaneously concentrated on the inner leaflet of the GUVs. They exhibited biased orientation and heterogeneous rotational dynamics as revealed by single particle rotationalmore » tracking. The combined experimental and simulation results show that Janus particles concentrate on the lipid membranes due to weak particle–lipid attraction, whereas the biased orientation of particles is driven predominantly by inter-particle interactions. Furthermore, this study demonstrates the potential of using lipid membranes to influence the self-assembly of Janus particles.« less

Simple estimation of Förster Resonance Energy Transfer (FRET) orientation factor distribution in membranes.

PubMed

Loura, Luís M S

2012-11-19

Because of its acute sensitivity to distance in the nanometer scale, Förster resonance energy transfer (FRET) has found a large variety of applications in many fields of chemistry, physics, and biology. One important issue regarding the correct usage of FRET is its dependence on the donor-acceptor relative orientation, expressed as the orientation factor k(2). Different donor/acceptor conformations can lead to k(2) values in the 0 ≤ k(2) ≤ 4 range. Because the characteristic distance for FRET, R(0), is proportional to (k(2))1/6, uncertainties in the orientation factor are reflected in the quality of information that can be retrieved from a FRET experiment. In most cases, the average value of k(2) corresponding to the dynamic isotropic limit ( = 2/3) is used for computation of R(0) and hence donor-acceptor distances and acceptor concentrations. However, this can lead to significant error in unfavorable cases. This issue is more critical in membrane systems, because of their intrinsically anisotropic nature and their reduced fluidity in comparison to most common solvents. Here, a simple numerical simulation method for estimation of the probability density function of k(2) for membrane-embedded donor and acceptor fluorophores in the dynamic regime is presented. In the simplest form, the proposed procedure uses as input the most probable orientations of the donor and acceptor transition dipoles, obtained by experimental (including linear dichroism) or theoretical (such as molecular dynamics simulation) techniques. Optionally, information about the widths of the donor and/or acceptor angular distributions may be incorporated. The methodology is illustrated for special limiting cases and common membrane FRET pairs.

Mode-selective mapping and control of vectorial nonlinear-optical processes in multimode photonic-crystal fibers.

PubMed

Hu, Ming-Lie; Wang, Ching-Yue; Song, You-Jian; Li, Yan-Feng; Chai, Lu; Serebryannikov, Evgenii; Zheltikov, Aleksei

2006-02-06

We demonstrate an experimental technique that allows a mapping of vectorial nonlinear-optical processes in multimode photonic-crystal fibers (PCFs). Spatial and polarization modes of PCFs are selectively excited in this technique by varying the tilt angle of the input beam and rotating the polarization of the input field. Intensity spectra of the PCF output plotted as a function of the input field power and polarization then yield mode-resolved maps of nonlinear-optical interactions in multimode PCFs, facilitating the analysis and control of nonlinear-optical transformations of ultrashort laser pulses in such fibers.

Characterization of the methotrexate transport pathway in murine L1210 leukemia cells: Involvement of a membrane receptor and a cytosolic protein

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

Price, E.M.; Ratnam, M.; Rodeman, K.M.

1988-10-04

A radioiodinated photoaffinity analogue of methotrexate, N{sup {alpha}}-(4-amino-4-deoxy-10-methyl-pteroyl)-N{sup {epsilon}}-(4-azidosalicylyl)-L-lysine (APA-ASA-Lys), was recently used to identify the plasma membrane derived binding protein involved in the transport of this folate antagonist into murine L1210 cells. The labeled protein has an apparent molecular weight of 46K-48K when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, but no such labeling occurs in a methotrexate transport-defective cell line (L1210/R81). Labeling of the total cytosolic protein from disrupted cells, followed by electrophoresis and autoradiography, showed, among other proteins, a 21K band, corresponding to dihydrofolate reductase (DHFR), in both the parent and R81 cells and a 38K bandmore » only in the parent cells. However, when whole cells were UV irradiated at various times at 37{degree}C following addition of radiolabeled APA-ASA-Lys, the 38K protein and DHFR were the only cytosolic proteins labeled in the parent cells, while the intact R81 cells showed no labeled cytosolic protein, since the photoprobe is not transported. Further, when the parent cells were treated with a pulse of radiolabeled photoprobe, followed by UV irradiation at different times at 37{degree}C, the probe appeared sequentially on the 48K membrane protein and both the 38K cytosolic protein and dihydrofolate reductase. A 48K protein could be detected in both parent L1210 cells and the R81 cells on Western blots using antisera to a membrane folate binding protein from human placenta. These results suggest a vectorial transport of APA-ASA-Lys or methotrexate and reduced folate coenzymes into murine L1210 cells mediated by a 48K integral membrane protein and a 38K cytosolic or peripheral membrane protein. The 38K protein may help in the trafficking of reduced folate coenzymes, shuttling them to various cytosolic targets.« less

Macroscopic Ensembles of Aligned Carbon Nanotubes in Bubble Imprints Studied by Polarized Raman Microscopy

DOE PAGES

Ushiba, Shota; Hoyt, Jordan; Masui, Kyoko; ...

2014-01-01

We study the alignment of single-wall carbon nanotubes (SWCNTs) in bubble imprints through polarized Raman microscopy. A hemispherical bubble containing SWCNTs is pressed against a glass substrate, resulting in an imprint of the bubble membrane with a coffee ring on the substrate. We find that macroscopic ensembles of aligned SWCNTs are obtained in the imprints, in which there are three patterns of orientations: (i) azimuthal alignment on the coffee ring, (ii) radial alignment at the edge of the membrane, and (iii) random orientation at the center of the membrane. We also find that the alignment of SWCNTs in the imprintsmore » can be manipulated by spinning bubbles. The orientation of SWCNTs on the coffee ring is directed radially, which is orthogonal to the case of unspun bubbles. This approach enables one to align SWCNTs in large quantities and in a short time, potentially opening up a wide range of CNT-based electronic and optical applications.« less

Arabidopsis  SABRE and CLASP interact to stabilize cell division plane orientation and planar polarity

PubMed Central

Pietra, Stefano; Gustavsson, Anna; Kiefer, Christian; Kalmbach, Lothar; Hörstedt, Per; Ikeda, Yoshihisa; Stepanova, Anna N.; Alonso, Jose M.; Grebe, Markus

2013-01-01

The orientation of cell division and the coordination of cell polarity within the plane of the tissue layer (planar polarity) contribute to shape diverse multicellular organisms. The root of Arabidopsis thaliana displays regularly oriented cell divisions, cell elongation and planar polarity providing a plant model system to study these processes. Here we report that the SABRE protein, which shares similarity with proteins of unknown function throughout eukaryotes, has important roles in orienting cell division and planar polarity. SABRE localizes at the plasma membrane, endomembranes, mitotic spindle and cell plate. SABRE stabilizes the orientation of CLASP-labelled preprophase band microtubules predicting the cell division plane, and of cortical microtubules driving cell elongation. During planar polarity establishment, sabre is epistatic to clasp at directing polar membrane domains of Rho-of-plant GTPases. Our findings mechanistically link SABRE to CLASP-dependent microtubule organization, shedding new light on the function of SABRE-related proteins in eukaryotes. PMID:24240534

Orientation and length of mammalian skeletal myocytes in response to a unidirectional stretch

NASA Technical Reports Server (NTRS)

Collinsworth, A. M.; Torgan, C. E.; Nagda, S. N.; Rajalingam, R. J.; Kraus, W. E.; Truskey, G. A.

2000-01-01

Effects of mechanical forces exerted on mammalian skeletal muscle cells during development were studied using an in vitro model to unidirectionally stretch cultured C2C12 cells grown on silastic membrane. Previous models to date have not studied these responses of the mammalian system specifically. The silastic membrane upon which these cells were grown exhibited linear strain behavior over the range of 3.6-14.6% strain, with a Poisson's ratio of approximately 0.5. To mimic murine in utero long bone growth, cell substrates were stretched at an average strain rate of 2.36%/day for 4 days or 1.77%/day for 6 days with an overall membrane strain of 9.5% and 10.6%, respectively. Both control and stretched fibers stained positively for the contractile protein, alpha-actinin, demonstrating muscle fiber development. An effect of stretch on orientation and length of myofibers was observed. At both strain rates, stretched fibers aligned at a smaller angle relative to the direction of stretch and were significantly longer compared to randomly oriented control fibers. There was no effect of duration of stretch on orientation or length, suggesting the cellular responses are independent of strain rate for the range tested. These results demonstrate that, under conditions simulating mammalian long bone growth, cultured myocytes respond to mechanical forces by lengthening and orienting along the direction of stretch.

Implicit membrane treatment of buried charged groups: application to peptide translocation across lipid bilayers.

PubMed

Lazaridis, Themis; Leveritt, John M; PeBenito, Leo

2014-09-01

The energetic cost of burying charged groups in the hydrophobic core of lipid bilayers has been controversial, with simulations giving higher estimates than certain experiments. Implicit membrane approaches are usually deemed too simplistic for this problem. Here we challenge this view. The free energy of transfer of amino acid side chains from water to the membrane center predicted by IMM1 is reasonably close to all-atom free energy calculations. The shape of the free energy profile, however, for the charged side chains needs to be modified to reflect the all-atom simulation findings (IMM1-LF). Membrane thinning is treated by combining simulations at different membrane widths with an estimate of membrane deformation free energy from elasticity theory. This approach is first tested on the voltage sensor and the isolated S4 helix of potassium channels. The voltage sensor is stably inserted in a transmembrane orientation for both the original and the modified model. The transmembrane orientation of the isolated S4 helix is unstable in the original model, but a stable local minimum in IMM1-LF, slightly higher in energy than the interfacial orientation. Peptide translocation is addressed by mapping the effective energy of the peptide as a function of vertical position and tilt angle, which allows identification of minimum energy pathways and transition states. The barriers computed for the S4 helix and other experimentally studied peptides are low enough for an observable rate. Thus, computational results and experimental studies on the membrane burial of peptide charged groups appear to be consistent. This article is part of a Special Issue entitled: Interfacially Active Peptides and Proteins. Guest Editors: William C. Wimley and Kalina Hristova. Copyright © 2014 Elsevier B.V. All rights reserved.

Monitoring the Orientational Changes of Alamethicin during Incorporation into Bilayer Lipid Membranes.

PubMed

Forbrig, Enrico; Staffa, Jana K; Salewski, Johannes; Mroginski, Maria Andrea; Hildebrandt, Peter; Kozuch, Jacek

2018-02-13

Antimicrobial peptides (AMPs) are the first line of defense after contact of an infectious invader, for example, bacterium or virus, with a host and an integral part of the innate immune system of humans. Their broad spectrum of biological functions ranges from cell membrane disruption over facilitation of chemotaxis to interaction with membrane-bound or intracellular receptors, thus providing novel strategies to overcome bacterial resistances. Especially, the clarification of the mechanisms and dynamics of AMP incorporation into bacterial membranes is of high interest, and different mechanistic models are still under discussion. In this work, we studied the incorporation of the peptaibol alamethicin (ALM) into tethered bilayer lipid membranes on electrodes in combination with surface-enhanced infrared absorption (SEIRA) spectroscopy. This approach allows monitoring the spontaneous and potential-induced ion channel formation of ALM in situ. The complex incorporation kinetics revealed a multistep mechanism that points to peptide-peptide interactions prior to penetrating the membrane and adopting the transmembrane configuration. On the basis of the anisotropy of the backbone amide I and II infrared absorptions determined by density functional theory calculations, we employed a mathematical model to evaluate ALM reorientations monitored by SEIRA spectroscopy. Accordingly, ALM was found to adopt inclination angles of ca. 69°-78° and 21° in its interfacially adsorbed and transmembrane incorporated states, respectively. These orientations can be stabilized efficiently by the dipolar interaction with lipid head groups or by the application of a potential gradient. The presented potential-controlled mechanistic study suggests an N-terminal integration of ALM into membranes as monomers or parallel oligomers to form ion channels composed of parallel-oriented helices, whereas antiparallel oligomers are barred from intrusion.

Membrane orientation and lateral diffusion of BODIPY-cholesterol as a function of probe structure.

PubMed

Solanko, Lukasz M; Honigmann, Alf; Midtiby, Henrik Skov; Lund, Frederik W; Brewer, Jonathan R; Dekaris, Vjekoslav; Bittman, Robert; Eggeling, Christian; Wüstner, Daniel

2013-11-05

Cholesterol tagged with the BODIPY fluorophore via the central difluoroboron moiety of the dye (B-Chol) is a promising probe for studying intracellular cholesterol dynamics. We synthesized a new BODIPY-cholesterol probe (B-P-Chol) with the fluorophore attached via one of its pyrrole rings to carbon-24 of cholesterol (B-P-Chol). Using two-photon fluorescence polarimetry in giant unilamellar vesicles and in the plasma membrane (PM) of living intact and actin-disrupted cells, we show that the BODIPY-groups in B-Chol and B-P-Chol are oriented perpendicular and almost parallel to the bilayer normal, respectively. B-Chol is in all three membrane systems much stronger oriented than B-P-Chol. Interestingly, we found that the lateral diffusion in the PM was two times slower for B-Chol than for B-P-Chol, although we found no difference in lateral diffusion in model membranes. Stimulated emission depletion microscopy, performed for the first time, to our knowledge, with fluorescent sterols, revealed that the difference in lateral diffusion of the BODIPY-cholesterol probes was not caused by anomalous subdiffusion, because diffusion of both analogs in the PM was free but not hindered. Our combined measurements show that the position and orientation of the BODIPY moiety in cholesterol analogs have a severe influence on lateral diffusion specifically in the PM of living cells. Copyright © 2013 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Mechanisms of Membrane Transport of Folates into Cells and Across Epithelia

PubMed Central

Zhao, Rongbao; Diop-Bove, Ndeye; Visentin, Michele; Goldman, I. David

2013-01-01

Until recently, the transport of folates into cells and across epithelia has been interpreted primarily within the context of two transporters with high affinity and specificity for folates, the reduced folate carrier and the folate receptors. However, there were discrepancies between the properties of these transporters and characteristics of folate transport in many tissues, most notably the intestinal absorption of folates, in terms of pH dependency and substrate specificity. With the recent cloning of the proton-coupled folate transporter (PCFT) and the demonstration that this transporter is mutated in hereditary folate malabsorption, an autosomal recessive disorder, the molecular basis for this low-pH transport activity is now understood. This review focuses on the properties of PCFT and briefly addresses the two other folate-specific transporters along with other facilitative and ATP-binding cassette (ABC) transporters with folate transport activities. The role of these transporters in the vectorial transport of folates across epithelia is considered. PMID:21568705

Mechanistic logic underlying the axonal transport of cytosolic proteins

PubMed Central

Scott, David A.; Das, Utpal; Tang, Yong; Roy, Subhojit

2011-01-01

Proteins vital to presynaptic function are synthesized in the neuronal perikarya and delivered into synapses via two modes of axonal transport. While membrane-anchoring proteins are conveyed in fast axonal transport via motor-driven vesicles, cytosolic proteins travel in slow axonal transport; via mechanisms that are poorly understood. We found that in cultured axons, populations of cytosolic proteins tagged to photoactivable-GFP (PA-GFP) move with a slow motor-dependent anterograde bias; distinct from vesicular-trafficking or diffusion of untagged PA-GFP. The overall bias is likely generated by an intricate particle-kinetics involving transient assembly and short-range vectorial spurts. In-vivo biochemical studies reveal that cytosolic proteins are organized into higher-order structures within axon-enriched fractions that are largely segregated from vesicles. Data-driven biophysical modeling best predicts a scenario where soluble molecules dynamically assemble into mobile supra-molecular structures. We propose a model where cytosolic proteins are transported by dynamically assembling into multi-protein complexes that are directly/indirectly conveyed by motors. PMID:21555071

Mechanical properties of electrospun bilayer fibrous membranes as potential scaffolds for tissue engineering.

PubMed

Pu, Juan; Komvopoulos, Kyriakos

2014-06-01

Bilayer fibrous membranes of poly(l-lactic acid) (PLLA) were fabricated by electrospinning, using a parallel-disk mandrel configuration that resulted in the sequential deposition of a layer with fibers aligned across the two parallel disks and a layer with randomly oriented fibers, both layers deposited in a single process step. Membrane structure and fiber alignment were characterized by scanning electron microscopy and two-dimensional fast Fourier transform. Because of the intricacies of the generated electric field, bilayer membranes exhibited higher porosity than single-layer membranes consisting of randomly oriented fibers fabricated with a solid-drum collector. However, despite their higher porosity, bilayer membranes demonstrated generally higher elastic modulus, yield strength and toughness than single-layer membranes with random fibers. Bilayer membrane deformation at relatively high strain rates comprised multiple abrupt microfracture events characterized by discontinuous fiber breakage. Bilayer membrane elongation yielded excessive necking of the layer with random fibers and remarkable fiber stretching (on the order of 400%) in the layer with fibers aligned in the stress direction. In addition, fibers in both layers exhibited multiple localized necking, attributed to the nonuniform distribution of crystalline phases in the fibrillar structure. The high membrane porosity, good mechanical properties, and good biocompatibility and biodegradability of PLLA (demonstrated in previous studies) make the present bilayer membranes good scaffold candidates for a wide range of tissue engineering applications. Copyright © 2014 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Theory of fluorescence polarization in magnetically oriented photosynthetic systems.

PubMed Central

Knox, R S; Davidovich, M A

1978-01-01

Many cells and cell fragments are known to assume specific alignments with respect to an applied magnetic field. One indicator of this alignment is a difference between the intensities of fluorescence observed in polarizations parallel and perpendicular to the magnetic filed. We calculate these two intensities using a model that assumes axially symmetric membranes and that covers a wide variety of shapes from flat disk to right cylinder. The fluorescence is assumed to originate at chromophores randomly exicted but nonrandomly oriented in the membranes. The membrane alignment is assumed to be due to the net torque on a nonrandom distribution of diamagnetically anisotropic molecules. The predicted results are consistent with most magnetoorientation data from green cells, but we are able to show that Chlorella data are not consistent with the hypothesis that the membranes have, and maintain, a cuplike configuration. Images FIGURE 4 FIGURE 5 PMID:737283

New insights into the targeting of a sub-set of tail-anchored proteins to the outer mitochondrial membrane

USDA-ARS?s Scientific Manuscript database

Tail-anchored (TA) proteins are a unique class of functionally diverse membrane proteins that are defined by their single C-terminal membrane-spanning domain and their ability to insert post-translationally into specific organelles with an Nout-Cin orientation. The molecular mechanisms by which TA p...

A post-classical theory of enamel biomineralization… and why we need one.

PubMed

Simmer, James P; Richardson, Amelia S; Hu, Yuan-Yuan; Smith, Charles E; Ching-Chun Hu, Jan

2012-09-01

Enamel crystals are unique in shape, orientation and organization. They are hundreds of thousands times longer than they are wide, run parallel to each other, are oriented with respect to the ameloblast membrane at the mineralization front and are organized into rod or interrod enamel. The classical theory of amelogenesis postulates that extracellular matrix proteins shape crystallites by specifically inhibiting ion deposition on the crystal sides, orient them by binding multiple crystallites and establish higher levels of crystal organization. Elements of the classical theory are supported in principle by in vitro studies; however, the classical theory does not explain how enamel forms in vivo. In this review, we describe how amelogenesis is highly integrated with ameloblast cell activities and how the shape, orientation and organization of enamel mineral ribbons are established by a mineralization front apparatus along the secretory surface of the ameloblast cell membrane.

C2 Domain of Protein Kinase Cα: Elucidation of the Membrane Docking Surface by Site-Directed Fluorescence and Spin Labeling†

PubMed Central

Kohout, Susy C.; Corbalán-García, Senena; Gómez-Fernández, Juan C.; Falke, Joseph J.

2013-01-01

The C2 domain is a conserved signaling motif that triggers membrane docking in a Ca2+-dependent manner, but the membrane docking surfaces of many C2 domains have not yet been identified. Two extreme models can be proposed for the docking of the protein kinase Cα (PKCα) C2 domain to membranes. In the parallel model, the membrane-docking surface includes the Ca2+ binding loops and an anion binding site on β-strands 3–4, such that the β-strands are oriented parallel to the membrane. In the perpendicular model, the docking surface is localized to the Ca2+ binding loops and the β-strands are oriented perpendicular to the membrane surface. The present study utilizes site-directed fluorescence and spin-labeling to map out the membrane docking surface of the PKCα C2 domain. Single cysteine residues were engineered into 18 locations scattered over all regions of the protein surface, and were used as attachment sites for spectroscopic probes. The environmentally sensitive fluorescein probe identified positions where Ca2+ activation or membrane docking trigger measurable fluorescence changes. Ca2+ binding was found to initiate a global conformational change, while membrane docking triggered the largest fluorescein environmental changes at labeling positions on the three Ca2+ binding loops (CBL), thereby localizing these loops to the membrane docking surface. Complementary EPR power saturation measurements were carried out using a nitroxide spin probe to determine a membrane depth parameter, Φ, for each spin-labeled mutant. Positive membrane depth parameters indicative of membrane insertion were found for three positions, all located on the Ca2+ binding loops: N189 on CBL 1, and both R249 and R252 on CBL 3. In addition, EPR power saturation revealed that five positions near the anion binding site are partially protected from collisions with an aqueous paramagnetic probe, indicating that the anion binding site lies at or near the surface of the headgroup layer. Together, the fluorescence and EPR results indicate that the Ca2+ first and third Ca2+ binding loops insert directly into the lipid headgroup region of the membrane, and that the anion binding site on β-strands 3–4 lies near the headgroups. The data support a model in which the β-strands are tilted toward the parallel orientation relative to the membrane surface. PMID:12564928

Method of fabricating electrode catalyst layers with directionally oriented carbon support for proton exchange membrane fuel cell

DOEpatents

Liu, Di-Jia [Naperville, IL; Yang, Junbing [Bolingbrook, IL

2012-03-20

A membrane electrode assembly (MEA) of the invention comprises an anode and a cathode and a proton conductive membrane therebetween, the anode and the cathode each comprising a patterned sheet of longitudinally aligned transition metal-containing carbon nanotubes, wherein the carbon nanotubes are in contact with and are aligned generally perpendicular to the membrane, wherein a catalytically active transition metal is incorporated throughout the nanotubes.

Architectured Nanomembranes

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

Sturgeon, Matthew R.; Hu, Michael Z.

2017-07-01

This paper has reviewed the frontier field of “architectured membranes” that contains anisotropic oriented porous nanostructures of inorganic materials. Three example types of architectured membranes were discussed with some relevant results from our own research: (1) anodized thin-layer titania membranes on porous anodized aluminum oxide (AAO) substrates of different pore sizes, (2) porous glass membranes on alumina substrate, and (3) guest-host membranes based on infiltration of yttrium-stabilized zirconia inside the pore channels of AAO matrices.

Large-scale fabrication of single crystalline tin nanowire arrays

NASA Astrophysics Data System (ADS)

Luo, Bin; Yang, Dachi; Liang, Minghui; Zhi, Linjie

2010-09-01

Large-scale single crystalline tin nanowire arrays with preferred lattice orientation along the [100] direction were fabricated in porous anodic aluminium oxide (AAO) membranes by the electrodeposition method using copper nanorod as a second electrode.Large-scale single crystalline tin nanowire arrays with preferred lattice orientation along the [100] direction were fabricated in porous anodic aluminium oxide (AAO) membranes by the electrodeposition method using copper nanorod as a second electrode. Electronic supplementary information (ESI) available: Experimental details and the information for single crystalline copper nanorods. See DOI: 10.1039/c0nr00206b

Rapid response to coastal upwelling in a semienclosed bay

NASA Astrophysics Data System (ADS)

Gilcoto, Miguel; Largier, John L.; Barton, Eric D.; Piedracoba, Silvia; Torres, Ricardo; Graña, Rocío.; Alonso-Pérez, Fernando; Villacieros-Robineau, Nicolás.; de la Granda, Francisco

2017-03-01

Bays/estuaries forced by local wind show bidirectional exchange flow. When forced by remote wind, they exhibit unidirectional flow adjustment to coastal sea level. Acoustic Doppler Current Profiler observations over 1 year show that the Ria de Vigo (Iberian Upwelling) responds to coastal wind events with bidirectional exchange flow. The duration of the upwelling and downwelling events, estimated from the current variability, was 3.3 days and 2.6 days, respectively. Vectorial correlations reveal a rapid response to upwelling/downwelling, in which currents lag local wind by <6 h and remote wind by <14 h, less than the Ekman spinup (17.8 h). This rapidity arises from the ria's narrowness (nonrotational local response), equatorward orientation (additive remote and local wind responses), depth greater than the Ekman depth (penetration of shelf circulation into the interior), and vertical stratification (shear reinforcing shelf circulation). Similar rapid responses are expected in other narrow bays where local and remote winds act together and stratification enhances bidirectional flow.

Membrane events in the acrosomal reaction of Limulus sperm. Membrane fusion, filament-membrane particle attachment, and the source and formation of new membrane surface

PubMed Central

1979-01-01

The membranes of Limulus (horseshoe crab) sperm were examined before and during the acrosomal reaction by using the technique of freeze- fracturing and thin sectioning. We focused on three areas. First, we examined stages in the fusion of the acrosomal vacuole with the cell surface. Fusion takes place in a particle-free zone which is surrounded by a circlet of particles on the P face of the plasma membrane and an underlying circlet of particles on the P face of the acrosomal vauole membrane. These circlets of particles are present before induction. Up to nine focal points of fusion occur within the particle-free zone. Second, we describe a system of fine filaments, each 30 A in diameter, which lies between the acrosomal vacuole and the plasma membrane. These filaments change their orientation as the vacuole opens, a process that takes place in less than 50 ms. Membrane particles seen on the P face of the acrosomal vacuole membrane change their orientation at the same time and in the same way as do the filaments, thus indicating that the membrane particles and filaments are probably connected. Third, we examined the source and the point of fusion of new membrane needed to cover the acrosomal process. This new membrane is almost certainly derived from the outer nuclear envelope and appears to insert into the plasma membrane in a particle-free area adjacent to an area rich in particles. The latter is the region where the particles are probably connected to the cytoplasmic filaments. The relevance of these observations in relation to the process of fertilization of this fantastic sperm is discussed. PMID:582596

Visualizing the kinetic power stroke that drives proton-coupled Zn(II) transport

PubMed Central

Gupta, Sayan; Chai, Jin; Cheng, Jie; D'Mello, Rhijuta; Chance, Mark R.; Fu, Dax

2014-01-01

The proton gradient is a principal energy source for respiration-dependent active transport, but the structural mechanisms of proton-coupled transport processes are poorly understood. YiiP is a proton-coupled zinc transporter found in the cytoplasmic membrane of E. coli, and the transport-site of YiiP receives protons from water molecules that gain access to its hydrophobic environment and transduces the energy of an inward proton gradient to drive Zn(II) efflux1,2. This membrane protein is a well characterized member3-7 of the protein family of cation diffusion facilitators (CDFs) that occurs at all phylogenetic levels8-10. X-ray mediated hydroxyl radical labeling of YiiP and mass spectrometric analysis showed that Zn(II) binding triggered a highly localized, all-or-none change of water accessibility to the transport-site and an adjacent hydrophobic gate. Millisecond time-resolved dynamics revealed a concerted and reciprocal pattern of accessibility changes along a transmembrane helix, suggesting a rigid-body helical reorientation linked to Zn(II) binding that triggers the closing of the hydrophobic gate. The gated water access to the transport-site enables a stationary proton gradient to facilitate the conversion of zinc binding energy to the kinetic power stroke of a vectorial zinc transport. The kinetic details provide energetic insights into a proton-coupled active transport reaction. PMID:25043033

Transmission-blocking antibodies against mosquito C-type lectins for dengue prevention.

PubMed

Liu, Yang; Zhang, Fuchun; Liu, Jianying; Xiao, Xiaoping; Zhang, Siyin; Qin, Chengfeng; Xiang, Ye; Wang, Penghua; Cheng, Gong

2014-02-01

C-type lectins are a family of proteins with carbohydrate-binding activity. Several C-type lectins in mammals or arthropods are employed as receptors or attachment factors to facilitate flavivirus invasion. We previously identified a C-type lectin in Aedes aegypti, designated as mosquito galactose specific C-type lectin-1 (mosGCTL-1), facilitating the attachment of West Nile virus (WNV) on the cell membrane. Here, we first identified that 9 A. aegypti mosGCTL genes were key susceptibility factors facilitating DENV-2 infection, of which mosGCTL-3 exhibited the most significant effect. We found that mosGCTL-3 was induced in mosquito tissues with DENV-2 infection, and that the protein interacted with DENV-2 surface envelop (E) protein and virions in vitro and in vivo. In addition, the other identified mosGCTLs interacted with the DENV-2 E protein, indicating that DENV may employ multiple mosGCTLs as ligands to promote the infection of vectors. The vectorial susceptibility factors that facilitate pathogen invasion may potentially be explored as a target to disrupt the acquisition of microbes from the vertebrate host. Indeed, membrane blood feeding of antisera against mosGCTLs dramatically reduced mosquito infective ratio. Hence, the immunization against mosGCTLs is a feasible approach for preventing dengue infection. Our study provides a future avenue for developing a transmission-blocking vaccine that interrupts the life cycle of dengue virus and reduces disease burden.

Microporous Poly(L-Lactic Acid) Membranes Fabricated by Polyethylene Glycol Solvent-Cast/Particulate Leaching Technique

PubMed Central

Selvam, Shivaram; Chang, Wenji V.; Nakamura, Tamako; Samant, Deedar M.; Thomas, Padmaja B.; Trousdale, Melvin D.; Mircheff, Austin K.; Schechter, Joel E.

2009-01-01

With the eventual goal of developing a tissue-engineered tear secretory system, we found that primary lacrimal gland acinar cells grown on solid poly(L-lactic acid) (PLLA) supports expressed the best histiotypic morphology. However, to be able to perform vectorial transport functions, epithelia must be supported by a permeable substratum. In the present study, we describe the use of a solvent-cast/particulate leaching technique to fabricate microporous PLLA membranes (mpPLLAm) from PLLA/polyethylene glycol blends. Scanning electron microscopy revealed pores on both the air-cured (∼4 μm) and glass-cured sides (<2 μm) of the mpPLLAm. Diffusion studies were performed with mpPLLAm fabricated from 57.1% PLLA/42.9% polyethylene glycol blends to confirm the presence of channelized pores. The data reveal that glucose, L-tryptophan, and dextran (a high molecular weight glucose polymer) readily permeate mpPLLAm. Diffusion of the immunoglobulin G through the mpPLLAm decreased with time, suggesting the possible adsorption and occlusion of the pores. Cells cultured on the mpPLLAm (57.1/42.9 wt%) grew to subconfluent monolayers but retained histiotypic morphological and physiological characteristics of lacrimal acinar cells in vivo. Our results suggest that mpPLLAm fabricated using this technique may be useful as a scaffold for a bioartificial lacrimal gland device. PMID:19260769

A Proteomic View at the Biochemistry of Syntrophic Butyrate Oxidation in Syntrophomonas wolfei

PubMed Central

Schmidt, Alexander; Müller, Nicolai; Schink, Bernhard; Schleheck, David

2013-01-01

In syntrophic conversion of butyrate to methane and CO2, butyrate is oxidized to acetate by secondary fermenting bacteria such as Syntrophomonas wolfei in close cooperation with methanogenic partner organisms, e.g., Methanospirillum hungatei. This process involves an energetically unfavourable shift of electrons from the level of butyryl-CoA oxidation to the substantially lower redox potential of proton and/or CO2 reduction, in order to transfer these electrons to the methanogenic partner via hydrogen and/or formate. In the present study, all prominent membrane-bound and soluble proteins expressed in S. wolfei specifically during syntrophic growth with butyrate, in comparison to pure-culture growth with crotonate, were examined by one- and two-dimensional gel electrophoresis, and identified by peptide fingerprinting-mass spectrometry. A membrane-bound, externally oriented, quinone-linked formate dehydrogenase complex was expressed at high level specifically during syntrophic butyrate oxidation, comprising a selenocystein-linked catalytic subunit with a membrane-translocation pathway signal (TAT), a membrane-bound iron-sulfur subunit, and a membrane-bound cytochrome. Soluble hydrogenases were expressed at high levels specifically during growth with crotonate. The results were confirmed by native protein gel electrophoresis, by formate dehydrogenase and hydrogenase-activity staining, and by analysis of formate dehydrogenase and hydrogenase activities in intact cells and cell extracts. Furthermore, constitutive expression of a membrane-bound, internally oriented iron-sulfur oxidoreductase (DUF224) was confirmed, together with expression of soluble electron-transfer flavoproteins (EtfAB) and two previously identified butyryl-CoA dehydrogenases. Our findings allow to depict an electron flow scheme for syntrophic butyrate oxidation in S. wolfei. Electrons derived from butyryl-CoA are transferred through a membrane-bound EtfAB:quinone oxidoreductase (DUF224) to a menaquinone cycle and further via a b-type cytochrome to an externally oriented formate dehydrogenase. Hence, an ATP hydrolysis-driven proton-motive force across the cytoplasmatic membrane would provide the energy input for the electron potential shift necessary for formate formation. PMID:23468890

Cell shape can mediate the spatial organization of the bacterial cytoskeleton

NASA Astrophysics Data System (ADS)

Wang, Siyuan; Wingreen, Ned

2013-03-01

The bacterial cytoskeleton guides the synthesis of cell wall and thus regulates cell shape. Since spatial patterning of the bacterial cytoskeleton is critical to the proper control of cell shape, it is important to ask how the cytoskeleton spatially self-organizes in the first place. In this work, we develop a quantitative model to account for the various spatial patterns adopted by bacterial cytoskeletal proteins, especially the orientation and length of cytoskeletal filaments such as FtsZ and MreB in rod-shaped cells. We show that the combined mechanical energy of membrane bending, membrane pinning, and filament bending of a membrane-attached cytoskeletal filament can be sufficient to prescribe orientation, e.g. circumferential for FtsZ or helical for MreB, with the accuracy of orientation increasing with the length of the cytoskeletal filament. Moreover, the mechanical energy can compete with the chemical energy of cytoskeletal polymerization to regulate filament length. Notably, we predict a conformational transition with increasing polymer length from smoothly curved to end-bent polymers. Finally, the mechanical energy also results in a mutual attraction among polymers on the same membrane, which could facilitate tight polymer spacing or bundling. The predictions of the model can be verified through genetic, microscopic, and microfluidic approaches.

Interactions of the Auxilin-1 PTEN-like Domain with Model Membranes Result in Nanoclustering of Phosphatidyl Inositol Phosphates

PubMed Central

Kalli, Antreas C.; Morgan, Gareth; Sansom, Mark S.P.

2013-01-01

Auxilin-1 is a neuron-specific membrane-binding protein involved in a late stage of clathrin-mediated endocytosis. It recruits Hsc70, thus initiating uncoating of the clathrin-coated vesicles. Interactions of auxilin-1 with the vesicle membrane are crucial for this function and are mediated via an N-terminal PTEN-like domain. We have used multiscale molecular dynamics simulations to probe the interactions of the auxilin-1 PTEN-like domain with lipid bilayers containing differing phospholipid composition, including bilayers containing phosphatidyl inositol phosphates. Our results suggest a novel, to our knowledge, model for the auxilin/membrane encounter and subsequent interactions. Negatively charged lipids (especially PIP2) enhance binding of auxilin to lipid bilayers and facilitate its correct orientation relative to the membrane. Mutations in three basic residues (R301E/R307E/K311E) of the C2 subdomain of the PTEN-like domain perturbed its interaction with the bilayer, changing its orientation. The interaction of membrane-bound auxilin-1 PTEN-like domain with negatively charged lipid headgroups results in nanoclustering of PIP2 molecules in the adjacent bilayer leaflet. PMID:23823232

Tailored vectorial light fields: flower, spider web and hybrid structures

NASA Astrophysics Data System (ADS)

Otte, Eileen; Alpmann, Christina; Denz, Cornelia

2017-04-01

We present the realization and analysis of tailored vector fields including polarization singularities. The fields are generated by a holographic method based on an advanced system including a spatial light modulator. We demonstrate our systems capabilities realizing specifically customized vector fields including stationary points of defined polarization in its transverse plane. Subsequently, vectorial flowers and spider webs as well as unique hybrid structures of these are introduced, and embedded singular points are characterized. These sophisticated light fields reveal attractive properties that pave the way to advanced application in e.g. optical micromanipulation. Beyond particle manipulation, they contribute essentially to actual questions in singular optics.

Formation of a Spin Texture in a Quantum Gas Coupled to a Cavity

NASA Astrophysics Data System (ADS)

Landini, M.; Dogra, N.; Kroeger, K.; Hruby, L.; Donner, T.; Esslinger, T.

2018-06-01

We observe cavity mediated spin-dependent interactions in an off-resonantly driven multilevel atomic Bose-Einstein condensate that is strongly coupled to an optical cavity. Applying a driving field with adjustable polarization, we identify the roles of the scalar and the vectorial components of the atomic polarizability tensor for single and multicomponent condensates. Beyond a critical strength of the vectorial coupling, we infer the formation of a spin texture in a condensate of two internal states from the analysis of the cavity output field. Our work provides perspectives for global dynamical gauge fields and self-consistently spin-orbit coupled gases.

A vectorial capacity product to monitor changing malaria transmission potential in epidemic regions of Africa

USGS Publications Warehouse

Ceccato, Pietro; Vancutsem, Christelle; Klaver, Robert; Rowland, James; Connor, Stephen J.

2012-01-01

Rainfall and temperature are two of the major factors triggering malaria epidemics in warm semi-arid (desert-fringe) and high altitude (highland-fringe) epidemic risk areas. The ability of the mosquitoes to transmit Plasmodium spp. is dependent upon a series of biological features generally referred to as vectorial capacity. In this study, the vectorial capacity model (VCAP) was expanded to include the influence of rainfall and temperature variables on malaria transmission potential. Data from two remote sensing products were used to monitor rainfall and temperature and were integrated into the VCAP model. The expanded model was tested in Eritrea and Madagascar to check the viability of the approach. The analysis of VCAP in relation to rainfall, temperature and malaria incidence data in these regions shows that the expanded VCAP correctly tracks the risk of malaria both in regions where rainfall is the limiting factor and in regions where temperature is the limiting factor. The VCAP maps are currently offered as an experimental resource for testing within Malaria Early Warning applications in epidemic prone regions of sub-Saharan Africa. User feedback is currently being collected in preparation for further evaluation and refinement of the VCAP model.

Geographical, meteorological and vectorial factors related to malaria re-emergence in Huang-Huai River of central China.

PubMed

Zhou, Shui S; Huang, Fang; Wang, Jian J; Zhang, Shao S; Su, Yun P; Tang, Lin H

2010-11-24

Malaria still represents a significant public health problem in China, and the cases dramatically increased in the areas along the Huang-Huai River of central China after 2001. Considering spatial aggregation of malaria cases and specific vectors, the geographical, meteorological and vectorial factors were analysed to determine the key factors related to malaria re-emergence in these particular areas. The geographic information of 357 malaria cases and 603 water bodies in 113 villages were collected to analyse the relationship between the residence of malaria cases and water body. Spearman rank correlation, multiple regression, curve fitting and trend analysis were used to explain the relationship between the meteorological factors and malaria incidence. Entomological investigation was conducted in two sites to get the vectorial capacity and the basic reproductive rate to determine whether the effect of vector lead to malaria re-emergence. The distances from household of cases to the nearest water-body was positive-skew distributed, the median was 60.9 m and 74% malaria cases were inhabited in the extent of 60 m near the water body, and the risk rate of people live there attacked by malaria was higher than others(OR = 1.6, 95%CI (1.042, 2.463), P < 0.05). The annual average temperature and rainfall may have close relationship with annual incidence. The average monthly temperature and rainfall were the key factors, and the correlation coefficients are 0.501 and 0.304(P < 0.01), respectively. Moreover, 75.3% changes of monthly malaria incidence contributed to the average monthly temperature (T(mean)), the average temperature of last two months(T(mean₀₁)) and the average rainfall of current month (R(mean)) and the regression equation was Y = -2.085 + 0.839I₁ + 0.998T(mean₀) - 0.86T(mean₀₁) + 0.16R(mean₀). All the collected mosquitoes were Anopheles sinensis. The vectorial capacity and the basic reproductive rate of An. sinensis in two sites were 0.6969, 0.4983 and 2.1604, 1.5447, respectively. The spatial distribution between malaria cases and water-body, the changing of meteorological factors, and increasing vectorial capacity and basic reproductive rate of An. sinensis leaded to malaria re-emergence in these areas.

Geographical, meteorological and vectorial factors related to malaria re-emergence in Huang-Huai River of central China

PubMed Central

2010-01-01

Background Malaria still represents a significant public health problem in China, and the cases dramatically increased in the areas along the Huang-Huai River of central China after 2001. Considering spatial aggregation of malaria cases and specific vectors, the geographical, meteorological and vectorial factors were analysed to determine the key factors related to malaria re-emergence in these particular areas. Methods The geographic information of 357 malaria cases and 603 water bodies in 113 villages were collected to analyse the relationship between the residence of malaria cases and water body. Spearman rank correlation, multiple regression, curve fitting and trend analysis were used to explain the relationship between the meteorological factors and malaria incidence. Entomological investigation was conducted in two sites to get the vectorial capacity and the basic reproductive rate to determine whether the effect of vector lead to malaria re-emergence. Results The distances from household of cases to the nearest water-body was positive-skew distributed, the median was 60.9 m and 74% malaria cases were inhabited in the extent of 60 m near the water body, and the risk rate of people live there attacked by malaria was higher than others(OR = 1.6, 95%CI (1.042, 2.463), P < 0.05). The annual average temperature and rainfall may have close relationship with annual incidence. The average monthly temperature and rainfall were the key factors, and the correlation coefficients are 0.501 and 0.304(P < 0.01), respectively. Moreover, 75.3% changes of monthly malaria incidence contributed to the average monthly temperature (Tmean), the average temperature of last two months(Tmean01) and the average rainfall of current month (Rmean) and the regression equation was Y = -2.085 + 0.839I1 + 0.998Tmean0 - 0.86Tmean01 + 0.16Rmean0. All the collected mosquitoes were Anopheles sinensis. The vectorial capacity and the basic reproductive rate of An. sinensis in two sites were 0.6969, 0.4983 and 2.1604, 1.5447, respectively. Conclusion The spatial distribution between malaria cases and water-body, the changing of meteorological factors, and increasing vectorial capacity and basic reproductive rate of An. sinensis leaded to malaria re-emergence in these areas. PMID:21092326

Topographical localization of the C-terminal region of the voltage-dependent sodium channel from Electrophorus electricus using antibodies raised against a synthetic peptide

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

Gordon, R.D.; Fieles, W.E.; Schotland, D.L.

1987-01-01

A peptide corresponding to amino acid residues 1783-1794 near the C terminus of the electric eel sodium channel primary sequence of the eel (Electrophorus electricus) sodium channel has been synthesized and used to raise an antiserum in rabbits. This antiserum specifically recognized the peptide in a solid-phase radioimmunoassay. Specificity of the antiserum for the native channel protein was shown by its specific binding to a 280-kDa protein in immunoblots of eel electroplax membrane proteins. The antiserum also specifically labeled the innervated membrane of the eel electroplax in immunofluorescent studies. The membrane topology of the peptide recognized by this antiserum wasmore » proved in binding studies using oriented electroplax membrane vesicles. These vesicles were 98% right-side-out as determined by (/sup 3/H)saxitoxin binding. Binding of the antipeptide antiserum to this fraction was measured before and after permeabilization with 0.01% saponin. Specific binding to intact vesicles was low, but this binding increased 10-fold after permeabilization, implying a cytoplasmic orientation for the peptide. Confirmation for this orientation was then sought by localizing the antibody bound to intact electroplax cells with immunogold electron microscopy. The data imply that the region of the sodium channel primary sequence near the C terminus that is recognized by the anitserum is localized on the cytoplasmic side of the membrane; this localization provides some further constraints on models of sodium channel tertiary structure.« less

Visualization of structural organization of ventral membranes of sheared-open resorbing osteoclasts attached to apatite pellets.

PubMed

Akisaka, Toshitaka; Yoshida, Atsushi

2015-05-01

Osteoclasts are highly polarized cells from both morphological and functional points of view. Using quick-freeze, rotary-replication methods combined with cell-shearing, we clarified the variability of cytoplasmic surface of the polarized membranes of osteoclasts seeded on apatite. As to the organization of actin filaments and clathrin sheets, we confirmed almost the same ventral membrane specializations of osteoclasts on apatite as seen on glass plates. The organized actin filaments and membrane-associated particles supported the ruffled border membranes. Inside the actin sealing zone, membrane specializations were not always occupied with the ruffled border but also with other types of membranes. Some osteoclasts formed an actin ring but lacked the ruffled border projections. We report a unique and distinctive membrane modification of apatite-attached osteoclasts, i.e., the presence of dense aggregates of membrane-associated particles and related structures not found in the osteoclasts seeded on glass plates. Actin filament polarity in the podosomes was determined by decoration with myosin S1. The actin filament polarity within podosome appears to be oriented predominantly with its barbed ends toward the core, whereas the interconnecting F-actin appears to be mixed oriented. Two different types of clathrin plaques displayed different distributions: clathrin-dependent endocytosis was observed in the ruffled border regions, whereas flat clathrin sheets were found in the leading edge of lamellipodia and near podosomes. The clathrin sheets adhered to the apatite surface tightly on the ventral membranes overlaying the resorption lacunae. All these membrane specializations as mentioned above may indicate the functional variability of osteoclasts seeded on apatite.

Structure and Membrane Interactions of the Antibiotic Peptide Dermadistinctin K by Multidimensional Solution and Oriented 15N and 31P Solid-State NMR Spectroscopy

PubMed Central

Verly, Rodrigo M.; Moraes, Cléria Mendonça de; Resende, Jarbas M.; Aisenbrey, Christopher; Bemquerer, Marcelo Porto; Piló-Veloso, Dorila; Valente, Ana Paula; Almeida, Fábio C.L.; Bechinger, Burkhard

2009-01-01

DD K, a peptide first isolated from the skin secretion of the Phyllomedusa distincta frog, has been prepared by solid-phase chemical peptide synthesis and its conformation was studied in trifluoroethanol/water as well as in the presence of sodium dodecyl sulfate and dodecylphosphocholine micelles or small unilamellar vesicles. Multidimensional solution NMR spectroscopy indicates an α-helical conformation in membrane environments starting at residue 7 and extending to the C-terminal carboxyamide. Furthermore, DD K has been labeled with 15N at a single alanine position that is located within the helical core region of the sequence. When reconstituted into oriented phosphatidylcholine membranes the resulting 15N solid-state NMR spectrum shows a well-defined helix alignment parallel to the membrane surface in excellent agreement with the amphipathic character of DD K. Proton-decoupled 31P solid-state NMR spectroscopy indicates that the peptide creates a high level of disorder at the level of the phospholipid headgroup suggesting that DD K partitions into the bilayer where it severely disrupts membrane packing. PMID:19289046

Reverse polarity magnetized melt rocks from the Chicxulub impact structure, Yucatan Peninsula, Mexico

NASA Astrophysics Data System (ADS)

Urrutia-Fucugauchi, Jaime; Marin, Luis E.; Sharpton, Virgil L.; Quezada, Juan Manuel

1993-03-01

Further paleomagnetic data for core samples of melt rock recovered in the Petroleos Mexicanos (PEMEX) exploratory wells within the Chicxulub structure, northern Yucatan peninsula, Mexico are reported. A previous report by Sharpton showed that the rocks studied contain high iridium levels and shocked breccia clasts, and an Ar-40/Ar-39 age of 65.2 plus or minus 0.4 Ma. The geomagnetic polarity determined for two samples is reverse (R) and was correlated with chron 29R that includes the K/T boundary. Our present analysis is based on two samples from each of three clasts of the melt rock from PEMEX well Y6-N17 (1295 to 1299 m b.s.l.). This study concentrates on the vectorial nature and stability of the remanence (NRM), the magnetic mineralogy and remanence carriers (i.e., the reliability and origin of the record), and on the implications (correlation with expected paleolatitude and polarity). The relative orientation of the drill core samples with respect to the horizontal is known. Samples were stable under alternating field (AF) and thermal treatments, and after removal of a small component they exhibited single-vectorial behavior. The characteristic remanence inclinations show small dispersion and a mean value (-43 deg) in close agreement with the expected inclination and paleolatitude (derived from the North American apparent polar wander path). Isothermal remenence (IRM) acquisition experiments, Lowrie-Fuller tests, coercivity and unblocking temperature spectra of NRM and saturation IRM, susceptibility and Q-coefficient analyses, and the single-component nature indicate a dominant mineralogy of iron-rich titanomagnetites with single or pseduo-single domain states. The stable characteristic magnetization may be interpreted as a result of shock heating of the rock at the time of formation of the inpact structure and its polarity, age, and paleolatitude are consistent with a time about the K/T boundary.

Reverse polarity magnetized melt rocks from the Chicxulub impact structure, Yucatan Peninsula, Mexico

NASA Technical Reports Server (NTRS)

Urrutia-Fucugauchi, Jaime; Marin, Luis E.; Sharpton, Virgil L.; Quezada, Juan Manuel

1993-01-01

Further paleomagnetic data for core samples of melt rock recovered in the Petroleos Mexicanos (PEMEX) exploratory wells within the Chicxulub structure, northern Yucatan peninsula, Mexico are reported. A previous report by Sharpton showed that the rocks studied contain high iridium levels and shocked breccia clasts, and an Ar-40/Ar-39 age of 65.2 plus or minus 0.4 Ma. The geomagnetic polarity determined for two samples is reverse (R) and was correlated with chron 29R that includes the K/T boundary. Our present analysis is based on two samples from each of three clasts of the melt rock from PEMEX well Y6-N17 (1295 to 1299 m b.s.l.). This study concentrates on the vectorial nature and stability of the remanence (NRM), the magnetic mineralogy and remanence carriers (i.e., the reliability and origin of the record), and on the implications (correlation with expected paleolatitude and polarity). The relative orientation of the drill core samples with respect to the horizontal is known. Samples were stable under alternating field (AF) and thermal treatments, and after removal of a small component they exhibited single-vectorial behavior. The characteristic remanence inclinations show small dispersion and a mean value (-43 deg) in close agreement with the expected inclination and paleolatitude (derived from the North American apparent polar wander path). Isothermal remenence (IRM) acquisition experiments, Lowrie-Fuller tests, coercivity and unblocking temperature spectra of NRM and saturation IRM, susceptibility and Q-coefficient analyses, and the single-component nature indicate a dominant mineralogy of iron-rich titanomagnetites with single or pseduo-single domain states. The stable characteristic magnetization may be interpreted as a result of shock heating of the rock at the time of formation of the inpact structure and its polarity, age, and paleolatitude are consistent with a time about the K/T boundary.

Single Particle Orientation and Rotational Tracking (SPORT) in biophysical studies

NASA Astrophysics Data System (ADS)

Gu, Yan; Ha, Ji Won; Augspurger, Ashley E.; Chen, Kuangcai; Zhu, Shaobin; Fang, Ning

2013-10-01

The single particle orientation and rotational tracking (SPORT) techniques have seen rapid development in the past 5 years. Recent technical advances have greatly expanded the applicability of SPORT in biophysical studies. In this feature article, we survey the current development of SPORT and discuss its potential applications in biophysics, including cellular membrane processes and intracellular transport.The single particle orientation and rotational tracking (SPORT) techniques have seen rapid development in the past 5 years. Recent technical advances have greatly expanded the applicability of SPORT in biophysical studies. In this feature article, we survey the current development of SPORT and discuss its potential applications in biophysics, including cellular membrane processes and intracellular transport. Electronic supplementary information (ESI) available: Three supplementary movies and an experimental section. See DOI: 10.1039/c3nr02254d

Microtubules Enable the Planar Cell Polarity of Airway Cilia

PubMed Central

Vladar, Eszter K.; Bayly, Roy D.; Sangoram, Ashvin; Scott, Matthew P.; Axelrod, Jeffrey D.

2012-01-01

Summary Background Airway cilia must be physically oriented along the longitudinal tissue axis for concerted, directional motility that is essential for proper mucociliary clearance. Results We show that Planar Cell Polarity (PCP) signaling specifies directionality and orients respiratory cilia. Within all airway epithelial cells a conserved set of PCP proteins shows interdependent, asymmetric junctional localization; non-autonomous signaling coordinates polarization between cells; and a polarized microtubule (MT) network is likely required for asymmetric PCP protein localization. We find that basal bodies dock after polarity of PCP proteins is established, are polarized nearly simultaneously, and refinement of basal body/cilium orientation continues during airway epithelial development. Unique to mature multiciliated cells, we identify PCP-regulated, planar polarized MTs that originate from basal bodies and interact, via their plus ends, with membrane domains associated with the PCP proteins Frizzled and Dishevelled. Disruption of MTs leads to misoriented cilia. Conclusions A conserved PCP pathway orients airway cilia by communicating polarity information from asymmetric membrane domains at the apical junctions, through MTs, to orient the MT and actin based network of ciliary basal bodies below the apical surface. PMID:23122850

Controlling the orientation of spin-correlated radical pairs by covalent linkage to nanoporous anodic aluminum oxide membranes.

PubMed

Chen, Hsiao-Fan; Gardner, Daniel M; Carmieli, Raanan; Wasielewski, Michael R

2013-10-07

Ordered multi-spin assemblies are required for developing solid-state molecule-based spintronics. A linear donor-chromophore-acceptor (D-C-A) molecule was covalently attached inside the 150 nm diam. nanopores of an anodic aluminum oxide (AAO) membrane. Photoexcitation of D-C-A in a 343 mT magnetic field results in sub-nanosecond, two-step electron transfer to yield the spin-correlated radical ion pair (SCRP) (1)(D(+)˙-C-A(-)˙), which then undergoes radical pair intersystem crossing (RP-ISC) to yield (3)(D(+)˙-C-A(-)˙). RP-ISC results in S-T0 mixing to selectively populate the coherent superposition states |S'> and |T'>. Microwave-induced transitions between these states and the unpopulated |T(+1)> and |T(-1)> states result in spin-polarized time-resolved EPR (TREPR) spectra. The dependence of the electron spin polarization (ESP) phase of the TREPR spectra on the orientation of the AAO membrane pores relative to the externally applied magnetic field is used to determine the overall orientation of the SCRPs within the pores at room temperature.

Small-amplitude backbone motions of the spin-labeled lipopeptide trichogin GA IV in a lipid membrane as revealed by electron spin echo.

PubMed

Syryamina, Victoria N; Isaev, Nikolay P; Peggion, Cristina; Formaggio, Fernando; Toniolo, Claudio; Raap, Jan; Dzuba, Sergei A

2010-09-30

Trichogin GA IV is a lipopeptide antibiotic of fungal origin, which is known to be able to modify the membrane permeability. TOAC nitroxide spin-labeled analogues of this membrane active peptide were investigated in hydrated bilayers of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) by electron spin echo (ESE) spectroscopy. Because the TOAC nitroxide spin label is rigidly attached to the peptide backbone, it may report on the backbone orientational dynamics. The ESE signal in this system is observed below ∼150 K. Previously, three-pulse stimulated ESE was found to be sensitive to two types of orientational motion of spin-labeled POPC lipid bilayers at these temperatures. The first type is fast stochastic librations, with a correlation time on the nanosecond scale (which also manifests itself in a two-pulse primary ESE experiment). The second type is slow millisecond inertial rotations. In the present work, we find that at low molar peptide to lipid ratio (1:200), where the individual peptide molecules are randomly distributed at the membrane surface, the spin labels show only a fast type of motion. At the high molar peptide to lipid ratio (1:20), a slow motion is also observed. Because at this high concentration trichogin GA IV is known to change its orientation from the in-plane topology to the transmembrane disposition, the observed onset of a slow motion may be safely attributed to the dynamics of peptides, which are elongated along the lipid molecules of the membrane. The possible interrelation between this backbone rotational motion of the peptide antibiotic and the membrane leakage is discussed.

Eicosapentaenoic acid and docosahexaenoic acid have distinct membrane locations and lipid interactions as determined by X-ray diffraction.

PubMed

Sherratt, Samuel C R; Mason, R Preston

2018-01-31

Eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) differentially influence lipid oxidation, signal transduction, fluidity, and cholesterol domain formation, potentially due in part to distinct membrane interactions. We used small angle X-ray diffraction to evaluate the EPA and DHA effects on membrane structure. Membrane vesicles composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and cholesterol (C) (0.3C:POPC mole ratio) were prepared and treated with vehicle, EPA, or DHA (1:10 mol ratio to POPC). Electron density profiles generated from the diffraction data showed that EPA increased membrane hydrocarbon core electron density over a broad area, up to ± 20 Å from the membrane center, indicating an energetically favorable extended orientation for EPA likely stabilized by van der Waals interactions. By contrast, DHA increased electron density in the phospholipid head group region starting at ± 12 Å from the membrane center, presumably due to DHA-surface interactions, with coincident reduction in electron density in the membrane hydrocarbon core centered ± 7-9 Å from the membrane center. The membrane width (d-space) decreased by 5 Å in the presence of vehicle as the temperature increased from 10 °C to 30 °C due to increased acyl chain trans-gauche isomerizations, which was unaffected by addition of EPA or DHA. The influence of DHA on membrane structure was modulated by temperature changes while the interactions of EPA were unaffected. The contrasting EPA and DHA effects on membrane structure indicate distinct molecular locations and orientations that may contribute to observed differences in biological activity. Copyright © 2018 The Authors. Published by Elsevier B.V. All rights reserved.

On the Reverse Asymmetric Gas Transport Effect in the Polymer Membranes

NASA Astrophysics Data System (ADS)

Kurchatov, I. M.; Laguntsov, N. I.; Skuridin, I. E.

In this paper, change of gas permeability value, depending on orientation of polymer gas membrane, in a wide pressure range was investigated. Consistent patterns of asymmetric gas transfer through the PVTMS-membrane were established experimentally. Reverse asymmetric transport effect was observed, wherein the permeability from the direction of porous support prevails at the permeability from the direction of selective non-porous layer.

Observing a model ion channel gating action in model cell membranes in real time in situ: membrane potential change induced alamethicin orientation change.

PubMed

Ye, Shuji; Li, Hongchun; Wei, Feng; Jasensky, Joshua; Boughton, Andrew P; Yang, Pei; Chen, Zhan

2012-04-11

Ion channels play crucial roles in transport and regulatory functions of living cells. Understanding the gating mechanisms of these channels is important to understanding and treating diseases that have been linked to ion channels. One potential model peptide for studying the mechanism of ion channel gating is alamethicin, which adopts a split α/3(10)-helix structure and responds to changes in electric potential. In this study, sum frequency generation vibrational spectroscopy (SFG-VS), supplemented by attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), has been applied to characterize interactions between alamethicin (a model for larger channel proteins) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) lipid bilayers in the presence of an electric potential across the membrane. The membrane potential difference was controlled by changing the pH of the solution in contact with the bilayer and was measured using fluorescence spectroscopy. The orientation angle of alamethicin in POPC lipid bilayers was then determined at different pH values using polarized SFG amide I spectra. Assuming that all molecules adopt the same orientation (a δ distribution), at pH = 6.7 the α-helix at the N-terminus and the 3(10)-helix at the C-terminus tilt at about 72° (θ(1)) and 50° (θ(2)) versus the surface normal, respectively. When pH increases to 11.9, θ(1) and θ(2) decrease to 56.5° and 45°, respectively. The δ distribution assumption was verified using a combination of SFG and ATR-FTIR measurements, which showed a quite narrow distribution in the angle of θ(1) for both pH conditions. This indicates that all alamethicin molecules at the surface adopt a nearly identical orientation in POPC lipid bilayers. The localized pH change in proximity to the bilayer modulates the membrane potential and thus induces a decrease in both the tilt and the bend angles of the two helices in alamethicin. This is the first reported application of SFG to the study of model ion channel gating mechanisms in model cell membranes. © 2012 American Chemical Society

The shape modulation of osteoblast-osteocyte transformation and its correlation with the fibrillar organization in secondary osteons: a SEM study employing the graded osmic maceration technique.

PubMed

Pazzaglia, Ugo E; Congiu, Terenzio; Marchese, Marcella; Dell'Orbo, Carlo

2010-06-01

Cortex fractured surface and graded osmic maceration techniques were used to study the secretory activity of osteoblasts, the transformation of osteoblast to osteocytes, and the structural organization of the matrix around the cells with scanning electron microscopy (SEM). A specialized membrane differentiation at the base of the cell was observed with finger-like, flattened processes which formed a diffuse meshwork. These findings suggested that this membrane differentiation below the cells had not only functioned in transporting collagen through the membrane but also in orienting the fibrils once assembled. Thin ramifications arose from the large and flat membrane foldings oriented perpendicular to the plane of the osteoblasts. This meshwork of fine filaments could not be visualized with SEM because they were obscured within the matrix substance. Their 3-D structure, however, should be similar to the canalicular system. The meshwork of large, flattened processes was no more evident in the cells which had completed their transformation into osteocytes.

Membrane Interactions of hIAPP Monomer and Oligomer with Lipid Membranes by Molecular Dynamics Simulations.

PubMed

Zhang, Mingzhen; Ren, Baiping; Liu, Yonglan; Liang, Guizhao; Sun, Yan; Xu, Lijian; Zheng, Jie

2017-08-16

Interaction of human islet amyloid polypeptide (hIAPP) peptides with cell membrane is crucial for the understanding of amyloid toxicity associated with Type II diabetes (T2D). While it is known that the hIAPP-membrane interactions are considered to promote hIAPP aggregation into fibrils and induce membrane disruption, the membrane-induced conformation, orientation, aggregation, and adsorption behaviors of hIAPP peptides have not been well understood at the atomic level. Herein, we perform all-atom explicit-water molecular dynamics (MD) simulations to study the adsorption, orientation, and surface interaction of hIAPP aggregates with different sizes (monomer to tetramer) and conformations (monomer with α-helix and tetramer with β-sheet-rich U-turn) upon adsorption on the lipid bilayers composed of both pure zwitterionic POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) and mixed anionic POPC/POPE (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine) (3:1) lipids. MD simulation results show that hIAPP monomer with α-helical conformation and hIAPP pentamer with β-sheet conformation can adsorb on both POPC and POPC/POPE bilayers via a preferential orientation of N-terminal residues facing toward the bilayer surface. The hIAPP aggregates show stronger interactions with mixed POPC/POPE lipids than pure POPC lipids, consistent with experimental observation that hIAPP adsorption and fibrililation are enhanced on mixed lipid bilayers. While electrostatic interactions are main attractive forces to drive the hIAPP aggregates to adsorb on both bilayers, the introduction of the more hydrophilic head groups of POPE lipids further promote the formation of the interfacial hydrogen bonds. Complement to our previous studies of hIAPP aggregates in bulk solution, this computational work increases our knowledge about the mechanism of amyloid peptide-membrane interactions that is central to the understanding the progression of all amyloid diseases.

An iterative algorithm for L1-TV constrained regularization in image restoration

NASA Astrophysics Data System (ADS)

Chen, K.; Loli Piccolomini, E.; Zama, F.

2015-11-01

We consider the problem of restoring blurred images affected by impulsive noise. The adopted method restores the images by solving a sequence of constrained minimization problems where the data fidelity function is the ℓ1 norm of the residual and the constraint, chosen as the image Total Variation, is automatically adapted to improve the quality of the restored images. Although this approach is general, we report here the case of vectorial images where the blurring model involves contributions from the different image channels (cross channel blur). A computationally convenient extension of the Total Variation function to vectorial images is used and the results reported show that this approach is efficient for recovering nearly optimal images.

Computational study of peptide permeation through membrane: searching for hidden slow variables

NASA Astrophysics Data System (ADS)

Cardenas, Alfredo E.; Elber, Ron

2013-12-01

Atomically detailed molecular dynamics trajectories in conjunction with Milestoning are used to analyse the different contributions of coarse variables to the permeation process of a small peptide (N-acetyl-l-tryptophanamide, NATA) through a 1,2-dioleoyl-sn-glycero-3-phosphocholine membrane. The peptide reverses its overall orientation as it permeates through the biological bilayer. The large change in orientation is investigated explicitly but is shown to impact the free energy landscape and permeation time only moderately. Nevertheless, a significant difference in permeation properties of the two halves of the membrane suggests the presence of other hidden slow variables. We speculate, based on calculation of the potential of mean force, that a conformational transition of NATA makes significant contribution to these differences. Other candidates for hidden slow variables may include water permeation and collective motions of phospholipids.

Interaction of hematoporphyrin with lipid membranes.

PubMed

Stępniewski, Michał; Kepczynski, Mariusz; Jamróz, Dorota; Nowakowska, Maria; Rissanen, Sami; Vattulainen, Ilpo; Róg, Tomasz

2012-04-26

Natural or synthetic porphyrins are being used as photosensitizers in photodiagnosis (PD) and photodynamic therapy (PDT) of malignancies and some other diseases. Understanding the interactions between porphyrins and cell membranes is therefore important to rationalize the uptake of photosensitizers and their passive transport through cell membranes. In this study, we consider the properties of hematoporphyrin (Hp), a well-known photosensitizer for PD and PDT, in the presence of a 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) bilayer that we use as a model system for protein-free cell membranes. For this purpose, we employed 200 ns atomic-scale molecular dynamics (MD) simulations for five systems containing the neutral (Hp(0)) or the dianionic form (Hp(2-)) of Hp and the POPC bilayer. MD simulations allowed one to estimate the position, orientation, and dynamics of Hp molecules inside the membrane. The dye molecules were found to reside in the phospholipid headgroup area close to the carbonyl groups of the POPC acyl chains. Their orientations were dependent on the protonation state of two propionic groups. Hp(2-) was found to have a lower affinity to enter the membrane than the neutral form. The dianions, being in the aqueous phase, formed stable dimers with a strictly determined geometry. Our results fully supported the experimental data and provide a more detailed molecular-level description of the interactions of photosensitizers with lipid membranes.

Determining the Topology of Integral Membrane Peptides Using EPR Spectroscopy

PubMed Central

Inbaraj, Johnson J.; Cardon, Thomas B.; Laryukhin, Mikhail; Grosser, Stuart M.

2008-01-01

This paper reports on the development of a new structural biology technique for determining the membrane topology of an integral membrane protein inserted into magnetically aligned phospholipid bilayers (bicelles) using EPR spectroscopy. The nitroxide spin probe, 2,2,6,6-tetramethylpiperidine-1-oxyl-4-amino-4-carboxylic acid (TOAC) was attached to the pore-lining transmembrane domain (M2δ) of the nicotinic acetylcholine receptor (AChR) and incorporated into a bicelle. The corresponding EPR spectra revealed hyperfine splittings that were highly dependent on the macroscopic orientation of the bicelles with respect to the static magnetic field. The helical tilt of the peptide can be easily calculated using the hyperfine splittings gleaned from the orientational dependent EPR spectra. A helical tilt of 14° was calculated for the M2δ peptide with respect to the bilayer normal of the membrane, which agrees well with previous 15N solid-state NMR studies. The helical tilt of the peptide was verified by simulating the corresponding EPR spectra using the standardized MOMD approach. This new method is advantageous because: (1) bicelle samples are easy to prepare, (2) the helical tilt can be directly calculated from the orientational-dependent hyperfine splitting in the EPR spectra, and (3) EPR spectroscopy is approximately 1000 fold more sensitive than 15N solid-state NMR spectroscopy; thus, the helical tilt of an integral membrane peptide can be determined with only 100 μg of peptide. The helical tilt can be determined more accurately by placing TOAC spin labels at several positions with this technique. PMID:16848493

The Role of Poly(Aspartic Acid) in the Precipitation of Calcium Phosphate in Confinement

PubMed Central

Cantaert, Bram; Beniash, Elia

2013-01-01

Many questions remain regarding the formation of ultrathin hydroxapatite (HAP) crystals within the confines of collagen fibrils of bones. These structures form through the interplay of the collagen matrix and non-collagenous proteins, and in vitro mineralization studies employing poly(aspartic acid) (PAsp) as a mimic of the non-collagenous proteins have generated mineralized fibrils with structures comparable to their biogenic counterparts. In this article, we employ the nanoscale cylindrical pores perforating track-etch filtration membranes to investigate the role of PAsp in controlling the infiltration and crystallization of calcium phosphate (CaP) within confined volumes. Oriented polycrystalline HAP and non-oriented octacalcium phosphate (OCP) rods precipitated within the membrane pores via an amorphous calcium phosphate (ACP) precursor, where PAsp increased the proportion of OCP rods. Further, ACP crystallized faster within the membranes than in bulk solution when PAsp was present, suggesting that PAsp inhibits crystallization in solution, but promotes it when bound to a substrate. Finally, in contrast to the collagen system, PAsp reduced the yield of intra-membrane mineral and failed to enhance infiltration. This suggests that a specific interaction between the collagen matrix and ACP/PAsp precursor particles drives effective infiltration. Thus, while orientation of HAP crystals can be achieved by confinement alone, the chemistry of the collagen matrix is necessary for efficient mineralisation with CaP. PMID:24409343

The Role of Poly(Aspartic Acid) in the Precipitation of Calcium Phosphate in Confinement.

PubMed

Cantaert, Bram; Beniash, Elia; Meldrum, Fiona C

2013-12-28

Many questions remain regarding the formation of ultrathin hydroxapatite (HAP) crystals within the confines of collagen fibrils of bones. These structures form through the interplay of the collagen matrix and non-collagenous proteins, and in vitro mineralization studies employing poly(aspartic acid) (PAsp) as a mimic of the non-collagenous proteins have generated mineralized fibrils with structures comparable to their biogenic counterparts. In this article, we employ the nanoscale cylindrical pores perforating track-etch filtration membranes to investigate the role of PAsp in controlling the infiltration and crystallization of calcium phosphate (CaP) within confined volumes. Oriented polycrystalline HAP and non-oriented octacalcium phosphate (OCP) rods precipitated within the membrane pores via an amorphous calcium phosphate (ACP) precursor, where PAsp increased the proportion of OCP rods. Further, ACP crystallized faster within the membranes than in bulk solution when PAsp was present, suggesting that PAsp inhibits crystallization in solution, but promotes it when bound to a substrate. Finally, in contrast to the collagen system, PAsp reduced the yield of intra-membrane mineral and failed to enhance infiltration. This suggests that a specific interaction between the collagen matrix and ACP/PAsp precursor particles drives effective infiltration. Thus, while orientation of HAP crystals can be achieved by confinement alone, the chemistry of the collagen matrix is necessary for efficient mineralisation with CaP.

Magnetization reversal in YIG/GGG(111) nanoheterostructures grown by laser molecular beam epitaxy.

PubMed

Krichevtsov, Boris B; Gastev, Sergei V; Suturin, Sergey M; Fedorov, Vladimir V; Korovin, Alexander M; Bursian, Viktor E; Banshchikov, Alexander G; Volkov, Mikhail P; Tabuchi, Masao; Sokolov, Nikolai S

2017-01-01

Thin (4-20 nm) yttrium iron garnet (Y 3 Fe 5 O 12 , YIG) layers have been grown on gadolinium gallium garnet (Gd 3 Ga 5 O 12 , GGG) 111-oriented substrates by laser molecular beam epitaxy in 700-1000 °C growth temperature range. The layers were found to have atomically flat step-and-terrace surface morphology with step height of 1.8 Å characteristic for YIG(111) surface. As the growth temperature is increased from 700 to 1000 °C the terraces become wider and the growth gradually changes from layer by layer to step-flow regime. Crystal structure studied by electron and X-ray diffraction showed that YIG lattice is co-oriented and laterally pseudomorphic to GGG with small rhombohedral distortion present perpendicular to the surface. Measurements of magnetic moment, magneto-optical polar and longitudinal Kerr effect (MOKE), and X-ray magnetic circular dichroism (XMCD) were used for study of magnetization reversal for different orientations of magnetic field. These methods and ferromagnetic resonance studies have shown that in zero magnetic field magnetization lies in the film plane due to both shape and induced anisotropies. Vectorial MOKE studies have revealed the presence of an in-plane easy magnetization axis. In-plane magnetization reversal was shown to occur through combination of reversible rotation and abrupt irreversible magnetization jump, the latter caused by domain wall nucleation and propagation. The field at which the flip takes place depends on the angle between the applied magnetic field and the easy magnetization axis and can be described by the modified Stoner-Wohlfarth model taking into account magnetic field dependence of the domain wall energy. Magnetization curves of individual tetrahedral and octahedral magnetic Fe 3+ sublattices were studied by XMCD.

Magnetization reversal in YIG/GGG(111) nanoheterostructures grown by laser molecular beam epitaxy

NASA Astrophysics Data System (ADS)

Krichevtsov, Boris B.; Gastev, Sergei V.; Suturin, Sergey M.; Fedorov, Vladimir V.; Korovin, Alexander M.; Bursian, Viktor E.; Banshchikov, Alexander G.; Volkov, Mikhail P.; Tabuchi, Masao; Sokolov, Nikolai S.

2017-12-01

Thin (4-20 nm) yttrium iron garnet (Y3Fe5O12, YIG) layers have been grown on gadolinium gallium garnet (Gd3Ga5O12, GGG) 111-oriented substrates by laser molecular beam epitaxy in 700-1000 °C growth temperature range. The layers were found to have atomically flat step-and-terrace surface morphology with step height of 1.8 Å characteristic for YIG(111) surface. As the growth temperature is increased from 700 to 1000 °C the terraces become wider and the growth gradually changes from layer by layer to step-flow regime. Crystal structure studied by electron and X-ray diffraction showed that YIG lattice is co-oriented and laterally pseudomorphic to GGG with small rhombohedral distortion present perpendicular to the surface. Measurements of magnetic moment, magneto-optical polar and longitudinal Kerr effect (MOKE), and X-ray magnetic circular dichroism (XMCD) were used for study of magnetization reversal for different orientations of magnetic field. These methods and ferromagnetic resonance studies have shown that in zero magnetic field magnetization lies in the film plane due to both shape and induced anisotropies. Vectorial MOKE studies have revealed the presence of an in-plane easy magnetization axis. In-plane magnetization reversal was shown to occur through combination of reversible rotation and abrupt irreversible magnetization jump, the latter caused by domain wall nucleation and propagation. The field at which the flip takes place depends on the angle between the applied magnetic field and the easy magnetization axis and can be described by the modified Stoner-Wohlfarth model taking into account magnetic field dependence of the domain wall energy. Magnetization curves of individual tetrahedral and octahedral magnetic Fe3+ sublattices were studied by XMCD.

Magnetization reversal in YIG/GGG(111) nanoheterostructures grown by laser molecular beam epitaxy

PubMed Central

Krichevtsov, Boris B.; Gastev, Sergei V.; Suturin, Sergey M.; Fedorov, Vladimir V.; Korovin, Alexander M.; Bursian, Viktor E.; Banshchikov, Alexander G.; Volkov, Mikhail P.; Tabuchi, Masao; Sokolov, Nikolai S.

2017-01-01

Abstract Thin (4–20 nm) yttrium iron garnet (Y3Fe5O12, YIG) layers have been grown on gadolinium gallium garnet (Gd3Ga5O12, GGG) 111-oriented substrates by laser molecular beam epitaxy in 700–1000 °C growth temperature range. The layers were found to have atomically flat step-and-terrace surface morphology with step height of 1.8 Å characteristic for YIG(111) surface. As the growth temperature is increased from 700 to 1000 °C the terraces become wider and the growth gradually changes from layer by layer to step-flow regime. Crystal structure studied by electron and X-ray diffraction showed that YIG lattice is co-oriented and laterally pseudomorphic to GGG with small rhombohedral distortion present perpendicular to the surface. Measurements of magnetic moment, magneto-optical polar and longitudinal Kerr effect (MOKE), and X-ray magnetic circular dichroism (XMCD) were used for study of magnetization reversal for different orientations of magnetic field. These methods and ferromagnetic resonance studies have shown that in zero magnetic field magnetization lies in the film plane due to both shape and induced anisotropies. Vectorial MOKE studies have revealed the presence of an in-plane easy magnetization axis. In-plane magnetization reversal was shown to occur through combination of reversible rotation and abrupt irreversible magnetization jump, the latter caused by domain wall nucleation and propagation. The field at which the flip takes place depends on the angle between the applied magnetic field and the easy magnetization axis and can be described by the modified Stoner–Wohlfarth model taking into account magnetic field dependence of the domain wall energy. Magnetization curves of individual tetrahedral and octahedral magnetic Fe3+ sublattices were studied by XMCD. PMID:28685003

Ion-/proton-conducting apparatus and method

DOEpatents

Yates, Matthew [Penfield, NY; Liu, Dongxia [Rochester, NY

2011-05-17

A c-axis-oriented HAP thin film synthesized by seeded growth on a palladium hydrogen membrane substrate. An exemplary synthetic process includes electrochemical seeding on the substrate, and secondary and tertiary hydrothermal treatments under conditions that favor growth along c-axes and a-axes in sequence. By adjusting corresponding synthetic conditions, an HAP this film can be grown to a controllable thickness with a dense coverage on the underlying substrate. The thin films have relatively high proton conductivity under hydrogen atmosphere and high temperature conditions. The c-axis oriented films may be integrated into fuel cells for application in the intermediate temperature range of 200-600.degree. C. The electrochemical-hydrothermal deposition technique may be applied to create other oriented crystal materials having optimized properties, useful for separations and catalysis as well as electronic and electrochemical applications, electrochemical membrane reactors, and in chemical sensors.

Insect Wing Membrane Topography Is Determined by the Dorsal Wing Epithelium

PubMed Central

Belalcazar, Andrea D.; Doyle, Kristy; Hogan, Justin; Neff, David; Collier, Simon

2013-01-01

The Drosophila wing consists of a transparent wing membrane supported by a network of wing veins. Previously, we have shown that the wing membrane cuticle is not flat but is organized into ridges that are the equivalent of one wing epithelial cell in width and multiple cells in length. These cuticle ridges have an anteroposterior orientation in the anterior wing and a proximodistal orientation in the posterior wing. The precise topography of the wing membrane is remarkable because it is a fusion of two independent cuticle contributions from the dorsal and ventral wing epithelia. Here, through morphological and genetic studies, we show that it is the dorsal wing epithelium that determines wing membrane topography. Specifically, we find that wing hair location and membrane topography are coordinated on the dorsal, but not ventral, surface of the wing. In addition, we find that altering Frizzled Planar Cell Polarity (i.e., Fz PCP) signaling in the dorsal wing epithelium alone changes the membrane topography of both dorsal and ventral wing surfaces. We also examined the wing morphology of two model Hymenopterans, the honeybee Apis mellifera and the parasitic wasp Nasonia vitripennis. In both cases, wing hair location and wing membrane topography are coordinated on the dorsal, but not ventral, wing surface, suggesting that the dorsal wing epithelium also controls wing topography in these species. Because phylogenomic studies have identified the Hymenotera as basal within the Endopterygota family tree, these findings suggest that this is a primitive insect character. PMID:23316434

A tectorin-based matrix and planar cell polarity genes are required for normal collagen-fibril orientation in the developing tectorial membrane.

PubMed

Goodyear, Richard J; Lu, Xiaowei; Deans, Michael R; Richardson, Guy P

2017-11-01

The tectorial membrane is an extracellular structure of the cochlea. It develops on the surface of the auditory epithelium and contains collagen fibrils embedded in a tectorin-based matrix. The collagen fibrils are oriented radially with an apically directed slant - a feature considered crucial for hearing. To determine how this pattern is generated, collagen-fibril formation was examined in mice lacking a tectorin-based matrix, epithelial cilia or the planar cell polarity genes Vangl2 and Ptk7 In wild-type mice, collagen-fibril bundles appear within a tectorin-based matrix at E15.5 and, as fibril number rapidly increases, become co-aligned and correctly oriented. Epithelial width measurements and data from Kif3a cKO mice suggest, respectively, that radial stretch and cilia play little, if any, role in determining normal collagen-fibril orientation; however, evidence from tectorin-knockout mice indicates that confinement is important. PRICKLE2 distribution reveals the planar cell polarity axis in the underlying epithelium is organised along the length of the cochlea and, in mice in which this polarity is disrupted, the apically directed collagen offset is no longer observed. These results highlight the importance of the tectorin-based matrix and epithelial signals for precise collagen organisation in the tectorial membrane. © 2017. Published by The Company of Biologists Ltd.

Selective excitation for spectral editing and assignment in separated local field experiments of oriented membrane proteins

NASA Astrophysics Data System (ADS)

Koroloff, Sophie N.; Nevzorov, Alexander A.

2017-01-01

Spectroscopic assignment of NMR spectra for oriented uniformly labeled membrane proteins embedded in their native-like bilayer environment is essential for their structure determination. However, sequence-specific assignment in oriented-sample (OS) NMR is often complicated by insufficient resolution and spectral crowding. Therefore, the assignment process is usually done by a laborious and expensive "shotgun" method involving multiple selective labeling of amino acid residues. Presented here is a strategy to overcome poor spectral resolution in crowded regions of 2D spectra by selecting resolved "seed" residues via soft Gaussian pulses inserted into spin-exchange separated local-field experiments. The Gaussian pulse places the selected polarization along the z-axis while dephasing the other signals before the evolution of the 1H-15N dipolar couplings. The transfer of magnetization is accomplished via mismatched Hartmann-Hahn conditions to the nearest-neighbor peaks via the proton bath. By optimizing the length and amplitude of the Gaussian pulse, one can also achieve a phase inversion of the closest peaks, thus providing an additional phase contrast. From the superposition of the selective spin-exchanged SAMPI4 onto the fully excited SAMPI4 spectrum, the 15N sites that are directly adjacent to the selectively excited residues can be easily identified, thereby providing a straightforward method for initiating the assignment process in oriented membrane proteins.

'Boomerang'-like insertion of a fusogenic peptide in a lipid membrane revealed by solid-state 19F NMR.

PubMed

Afonin, Sergii; Dürr, Ulrich H N; Glaser, Ralf W; Ulrich, Anne S

2004-02-01

Solid state (19)F NMR revealed the conformation and alignment of the fusogenic peptide sequence B18 from the sea urchin fertilization protein bindin embedded in flat phospholipid bilayers. Single (19)F labels were introduced into nine distinct positions along the wild-type sequence by substituting each hydrophobic amino acid, one by one, with L-4-fluorophenylglycine. Their anisotropic chemical shifts were measured in uniaxially oriented membrane samples and used as orientational constraints to model the peptide structure in the membrane-bound state. Previous (1)H NMR studies of B18 in 30% TFE and in detergent micelles had shown that the peptide structure consists of two alpha-helical segments that are connected by a flexible hinge. This helix-break-helix motif was confirmed here by the solid-state (19)F NMR data, while no other secondary structure (beta-sheet, 3(10)-helix) was compatible with the set of orientational constraints. For both alpha-helical segments we found that the helical conformation extends all the way to the respective N- and C-termini of the peptide. Analysis of the corresponding tilt and azimuthal rotation angles showed that the N-terminal helix of B18 is immersed obliquely into the bilayer (at a tilt angle tau approximately 54 degrees), whereas the C-terminus is peripherally aligned (tau approximately 91 degrees). The azimuthal orientation of the two segments is consistent with the amphiphilic distribution of side-chains. The observed 'boomerang'-like mode of insertion into the membrane may thus explain how peptide binding leads to lipid dehydration and acyl chain perturbation as a prerequisite for bilayer fusion to occur. Copyright 2004 John Wiley & Sons, Ltd.

Direct observation of temperature-driven magnetic symmetry transitions by vectorial resolved MOKE magnetometry

NASA Astrophysics Data System (ADS)

Cuñado, Jose Luis F.; Pedrosa, Javier; Ajejas, Fernando; Perna, Paolo; Miranda, Rodolfo; Camarero, Julio

2017-10-01

Angle- and temperature-dependent vectorial magnetometry measurements are necessary to disentangle the effective magnetic symmetry in magnetic nanostructures. Here we present a detailed study on an Fe(1 0 0) thin film system with competing collinear biaxial (four-fold symmetry) and uniaxial (two-fold) magnetic anisotropies, carried out with our recently developed full angular/broad temperature range/vectorial-resolved magneto-optical Kerr effect magnetometer, named TRISTAN. The data give direct views on the angular and temperature dependence of the magnetization reversal pathways, from which characteristic axes, remanences, critical fields, domain wall types, and effective magnetic symmetry are obtained. In particular, although the remanence shows four-fold angular symmetry for all investigated temperatures (15 K-400 K), the critical fields show strong temperature and angular dependencies and the reversal mechanism changes for specific angles at a given (angle-dependent) critical temperature, showing signatures of an additional collinear two-fold symmetry. This symmetry-breaking is more relevant as temperature increases to room temperature. It originates from the competition between two anisotropy contributions with different symmetry and temperature evolution. The results highlight the importance of combining temperature and angular studies, and the need to look at different magnetic parameters to unravel the underlying magnetic symmetries and temperature evolutions of the symmetry-breaking effects in magnetic nanostructures.

Three-dimensional characterization of tightly focused fields for various polarization incident beams

NASA Astrophysics Data System (ADS)

Cai, Yanan; Liang, Yansheng; Lei, Ming; Yan, Shaohui; Wang, Zhaojun; Yu, Xianghua; Li, Manman; Dan, Dan; Qian, Jia; Yao, Baoli

2017-06-01

Tightly focused vectorial optical beams have found extensive applications in variety of technical fields like single-molecule detection, optical tweezers, and super-resolution optical microscopy. Such applications require an accurate measurement and manipulation of focal optical fields. We have developed a compact instrument (with dimensions of 35 × 35 × 30 cm3) to rapidly measure the intensity distribution in three dimensions of the focused fields of vectorial beams and any other incident beams. This instrument employs a fluorescent nanoparticle as a probe to scan the focal region to obtain a high spatial resolution of intensity distribution. It integrates a liquid-crystal spatial light modulator to allow for tailoring the point spread function of the optical system, making it a useful tool for multi-purpose and flexible research. The robust applicability of the instrument is verified by measuring the 3D intensity distributions of focal fields of various polarization and wavefront modulated incident beams focused by a high NA (=1.25) objective lens. The minimal data acquisition time achievable in the experiment is about 8 s for a scanning region of 3.2 × 3.2 μm2 (512 × 512 pixels). The measured results are in good agreement with those predicted by the vectorial diffraction theory.

integrating Solid State NMR and Computations in Membrane Protein Science

NASA Astrophysics Data System (ADS)

Cross, Timothy

2015-03-01

Helical membrane protein structures are influenced by their native environment. Therefore the characterization of their structure in an environment that models as closely as possible their native environment is critical for achieving not only structural but functional understanding of these proteins. Solid state NMR spectroscopy in liquid crystalline lipid bilayers provides an excellent tool for such characterizations. Two classes of restraints can be obtained - absolute restraints that constrain the structure to a laboratory frame of reference when using uniformly oriented samples (approximately 1° of mosaic spread) and relative restraints that restrain one part of the structure with respect to another part such as torsional and distance restraints. Here, I will discuss unique restraints derived from uniformly oriented samples and the characterization of initial structures utilizing both restraint types, followed by restrained molecular dynamics refinement in the same lipid bilayer environment as that used for the experimental restraint collection. Protein examples will be taken from Influenza virus and Mycobacterium tuberculosis. When available comparisons of structures to those obtained using different membrane mimetic environments will be shown and the causes for structural distortions explained based on an understanding of membrane biophysics and its sophisticated influence on membrane proteins.

Structural insights of a self-assembling 9-residue peptide from the C-terminal tail of the SARS corona virus E-protein in DPC and SDS micelles: A combined high and low resolution spectroscopic study.

PubMed

Ghosh, Anirban; Bhattacharyya, Dipita; Bhunia, Anirban

2018-02-01

In recent years, several studies based on the interaction of self-assembling short peptides derived from viroporins with model membranes, have improved our understanding of the molecular mechanism of corona virus (CoV) infection under physiological conditions. In this study, we have characterized the mechanism of membrane interaction of a short, 9-residue peptide TK9 (T 55 VYVYSRVK 63 ) that had been derived from the carboxyl terminal of the Severe Acute Respiratory Syndrome (SARS) corona virus (SARS CoV) envelope (E) protein. The peptide has been studied for its physical changes in the presence of both zwitterionic DPC and negatively charged SDS model membrane micelles, respectively, with the help of a battery of biophysical techniques including two-dimensional solution state NMR spectroscopy. Interestingly, in both micellar environments, TK9 adopted an alpha helical conformation; however, the helical propensities were much higher in the case of DPC compared to those of SDS micelle, suggesting that TK9 has more specificity towards eukaryotic cell membrane than the bacterial cell membrane. The orientation of the peptide TK9 also varies in the different micellar environments. The peptide's affinity was further manifested by its pronounced membrane disruption ability towards the mammalian compared to the bacterial membrane mimic. Collectively, the in-depth structural information on the interaction of TK9 with different membrane environments explains the host specificity and membrane orientation owing to subsequent membrane disruption implicated in the viral pathogenesis. Copyright © 2017 Elsevier B.V. All rights reserved.

Dynamic nuclear polarization methods in solids and solutions to explore membrane proteins and membrane systems.

PubMed

Cheng, Chi-Yuan; Han, Songi

2013-01-01

Membrane proteins regulate vital cellular processes, including signaling, ion transport, and vesicular trafficking. Obtaining experimental access to their structures, conformational fluctuations, orientations, locations, and hydration in membrane environments, as well as the lipid membrane properties, is critical to understanding their functions. Dynamic nuclear polarization (DNP) of frozen solids can dramatically boost the sensitivity of current solid-state nuclear magnetic resonance tools to enhance access to membrane protein structures in native membrane environments. Overhauser DNP in the solution state can map out the local and site-specific hydration dynamics landscape of membrane proteins and lipid membranes, critically complementing the structural and dynamics information obtained by electron paramagnetic resonance spectroscopy. Here, we provide an overview of how DNP methods in solids and solutions can significantly increase our understanding of membrane protein structures, dynamics, functions, and hydration in complex biological membrane environments.

Optimization of bicelle lipid composition and temperature for EPR spectroscopy of aligned membranes.

PubMed

McCaffrey, Jesse E; James, Zachary M; Thomas, David D

2015-01-01

We have optimized the magnetic alignment of phospholipid bilayered micelles (bicelles) for EPR spectroscopy, by varying lipid composition and temperature. Bicelles have been extensively used in NMR spectroscopy for several decades, in order to obtain aligned samples in a near-native membrane environment and take advantage of the intrinsic sensitivity of magnetic resonance to molecular orientation. Recently, bicelles have also seen increasing use in EPR, which offers superior sensitivity and orientational resolution. However, the low magnetic field strength (less than 1 T) of most conventional EPR spectrometers results in homogeneously oriented bicelles only at a temperature well above physiological. To optimize bicelle composition for magnetic alignment at reduced temperature, we prepared bicelles containing varying ratios of saturated (DMPC) and unsaturated (POPC) phospholipids, using EPR spectra of a spin-labeled fatty acid to assess alignment as a function of lipid composition and temperature. Spectral analysis showed that bicelles containing an equimolar mixture of DMPC and POPC homogeneously align at 298 K, 20 K lower than conventional DMPC-only bicelles. It is now possible to perform EPR studies of membrane protein structure and dynamics in well-aligned bicelles at physiological temperatures and below. Copyright © 2014 Elsevier Inc. All rights reserved.

Simultaneous acquisition of 2D and 3D solid-state NMR experiments for sequential assignment of oriented membrane protein samples.

PubMed

Gopinath, T; Mote, Kaustubh R; Veglia, Gianluigi

2015-05-01

We present a new method called DAISY (Dual Acquisition orIented ssNMR spectroScopY) for the simultaneous acquisition of 2D and 3D oriented solid-state NMR experiments for membrane proteins reconstituted in mechanically or magnetically aligned lipid bilayers. DAISY utilizes dual acquisition of sine and cosine dipolar or chemical shift coherences and long living (15)N longitudinal polarization to obtain two multi-dimensional spectra, simultaneously. In these new experiments, the first acquisition gives the polarization inversion spin exchange at the magic angle (PISEMA) or heteronuclear correlation (HETCOR) spectra, the second acquisition gives PISEMA-mixing or HETCOR-mixing spectra, where the mixing element enables inter-residue correlations through (15)N-(15)N homonuclear polarization transfer. The analysis of the two 2D spectra (first and second acquisitions) enables one to distinguish (15)N-(15)N inter-residue correlations for sequential assignment of membrane proteins. DAISY can be implemented in 3D experiments that include the polarization inversion spin exchange at magic angle via I spin coherence (PISEMAI) sequence, as we show for the simultaneous acquisition of 3D PISEMAI-HETCOR and 3D PISEMAI-HETCOR-mixing experiments.

Effects of infectious virus dose and bloodmeal delivery method on susceptibility of Aedes aegypti and Aedes albopictus to chikungunya virus.

PubMed

Pesko, Kendra; Westbrook, Catherine J; Mores, Christopher N; Lounibos, L Philip; Reiskind, Michael H

2009-03-01

Chikungunya virus (CHIKV) is an arbovirus (genus Alphavirus, family Togaviridae) that has recently caused disease outbreaks in the Indian Ocean basin and southern Europe. These outbreaks could be associated with a possible shift in primary vector from Aedes aegypti to Ae. albopictus. To evaluate vector competence differences in possible CHIKV vectors, we evaluated the dose-dependant susceptibility of Florida strains of Ae. albopictus and Ae. aegypti for infection with a La Réunion island strain of CHIKV. Pledget and water-jacketed membrane feeding systems were also evaluated. We show that both Aedes spp. were susceptible to the highest CHIKV doses, whereas only Ae. albopictus developed disseminated infections after exposure to the two lowest doses. Infection rates for both mosquito species were significantly affected by the bloodmeal delivery method used. This information is important in assessing risk of an outbreak of imported CHIKV in the United States, in determining differences in vectorial capacity of these two vector species, and in evaluating arbovirus delivery methods in the laboratory.

Effects of Infectious Virus Dose and Bloodmeal Delivery Method on Susceptibility of Aedes aegypti and Aedes albopictus to Chikungunya Virus

PubMed Central

Pesko, Kendra; Westbrook, Catherine J.; Mores, Christopher N.; Lounibos, L. Philip; Reiskind, Michael H.

2009-01-01

Chikungunya virus (CHIKV) is an arbovirus (genus Alphavirus, family Togaviridae) that has recently caused disease outbreaks in the Indian Ocean basin and southern Europe. These outbreaks could be associated with a possible shift in primary vector from Aedes aegypti to Ae. albopictus. To evaluate vector competence differences in possible CHIKV vectors, we evaluated the dose-dependant susceptibility of Florida strains of Ae. albopictus and Ae. aegypti for infection with a La Réunion island strain of CHIKV. Pledget and water-jacketed membrane feeding systems were also evaluated. We show that both Aedes spp. were susceptible to the highest CHIKV doses, whereas only Ae. albopictus developed disseminated infections after exposure to the two lowest doses. Infection rates for both mosquito species were significantly affected by the bloodmeal delivery method used. This information is important in assessing risk of an outbreak of imported CHIKV in the United States, in determining differences in vectorial capacity of these two vector species, and in evaluating arbovirus delivery methods in the laboratory. PMID:19351094

Comparative vectorial efficiency of Lutzomyia evansi and Lu. longipalpis for transmitting Leishmania chagasi.

PubMed

Montoya-Lerma, J; Cadena, H; Oviedo, M; Ready, P D; Barazarte, R; Travi, B L; Lane, R P

2003-01-01

The infection rates and development of Leishmania chagasi in two sandfly species, Lutzomyia evansi and Lutzomyia longipalpis, were evaluated under natural and experimental conditions. Natural infection rates of Lu. evansi in San Andrés de Sotavento (Colombia) and Montañas de Peraza (Venezuela) (0.05 and 0.2%, respectively) were similar to those previously recorded for this species in Colombia and Venezuela and for Lu. longipalpis in many foci of American Visceral Leishmaniasis (AVL). Both sand fly species were able to support the development of two Colombian strains of L. chagasi experimentally acquired from dogs, hamsters or membrane feeders. However, the experimental infection rates and the sequence of parasite development in the guts of these sand flies revealed that parasite colonisation, differentiation, migration and attachment were more frequent and uniform in Lu. longipalpis than in Lu. evansi. This is consistent with a more recent association between L. chagasi and Lu. evansi, and these results might help to explain the irregularity of AVL outbreaks in foci where Lu. evansi has been reported as the sole vector. Copyright 2002 Elsevier Science B.V.

Rapid pH change due to bacteriorhodopsin measured with a tin-oxide electrode.

PubMed Central

Robertson, B; Lukashev, E P

1995-01-01

The photocurrent transient generated by bacteriorhodopsin (bR) on a tin-oxide electrode is due to pH change and not to charge displacement as previously assumed. Films of either randomly oriented or highly oriented purple membranes were deposited on transparent electrodes made of tin-oxide-coated glass. The membranes contained either wild-type or D96N-mutant bR. When excited with yellow light through the glass, the bR pumps protons across the membrane. The result is a rapid local pH change as well as a charge displacement. Experiments with these films show that it is the pH change rather than the displacement that produces the current transient. The calibration for the transient pH measurement is given. The sensitivity of a tin-oxide electrode to a transient pH change is very much larger than its sensitivity to a steady-state pH change. PMID:7787036

Temporal correlations in the Vicsek model with vectorial noise

NASA Astrophysics Data System (ADS)

Gulich, Damián; Baglietto, Gabriel; Rozenfeld, Alejandro F.

2018-07-01

We study the temporal correlations in the evolution of the order parameter ϕ(t) for the Vicsek model with vectorial noise by estimating its Hurst exponent H with detrended fluctuation analysis (DFA). We present results on this parameter as a function of noise amplitude η introduced in simulations. We also compare with well known order-disorder phase transition for that same noise range. We find that - regardless of detrending degree - H spikes at the known coexistence noise for phase transition, and that this is due to nonstationarities introduced by the transit of the system between two well defined states with lower exponents. We statistically support this claim by successfully synthesizing equivalent cases derived from a transformed fractional Brownian motion (TfBm).

Vectorial Modeling Of NH In Comet 2P/Encke

NASA Astrophysics Data System (ADS)

Dorman, Garrett; Pierce, D.; Cochran, A.

2010-10-01

Encke is an ideal comet for studying the relationship of radicals to their photodissociative parent molecules due to its low dust content. On 2003 October 22 - 24, we used the the 2.7 m telescope at the McDonald Observatory of the University of Texas to obtain spectra of several cometary radical species. Using a version of the Vectorial Model that has been modified to simulate Encke's prominent sunward-facing fan, we examined the spacial distribution of NH in the coma. Potential photochemical parents of NH were studied in order to understand its production and spacial distribution in the coma. Derived production rates are compared to values in other comets to constrain the primary parent of NH in Encke.

Rigorous simulations of a helical core fiber by the use of transformation optics formalism.

PubMed

Napiorkowski, Maciej; Urbanczyk, Waclaw

2014-09-22

We report for the first time on rigorous numerical simulations of a helical-core fiber by using a full vectorial method based on the transformation optics formalism. We modeled the dependence of circular birefringence of the fundamental mode on the helix pitch and analyzed the effect of a birefringence increase caused by the mode displacement induced by a core twist. Furthermore, we analyzed the complex field evolution versus the helix pitch in the first order modes, including polarization and intensity distribution. Finally, we show that the use of the rigorous vectorial method allows to better predict the confinement loss of the guided modes compared to approximate methods based on equivalent in-plane bending models.

Ion-conducting ceramic apparatus, method, fabrication, and applications

DOEpatents

Yates, Matthew [Penfield, NY; Liu, Dongxia [Rochester, NY

2012-03-06

A c-axis-oriented HAP thin film synthesized by seeded growth on a palladium hydrogen membrane substrate. An exemplary synthetic process includes electrochemical seeding on the substrate, and secondary and tertiary hydrothermal treatments under conditions that favor growth along c-axes and a-axes in sequence. By adjusting corresponding synthetic conditions, an HAP this film can be grown to a controllable thickness with a dense coverage on the underlying substrate. The thin films have relatively high proton conductivity under hydrogen atmosphere and high temperature conditions. The c-axis oriented films may be integrated into fuel cells for application in the intermediate temperature range of 200-600.degree. C. The electrochemical-hydrothermal deposition technique may be applied to create other oriented crystal materials having optimized properties, useful for separations and catalysis as well as electronic and electrochemical applications, electrochemical membrane reactors, and in chemical sensors.

Improved monitoring of oriental fruit moth (Lepidoptera: Tortricidae) with terpinyl acetate plus acetic acid membrane lures

USDA-ARS?s Scientific Manuscript database

Male and female moth catches of Grapholita molesta (Busck) in traps were evaluated in stone and pome fruit orchards untreated or treated with sex pheromones for mating disruption in Uruguay, Argentina, Chile, USA, and Italy from 2015 - 2017. Trials evaluated various blends loaded into either membran...

Location and ion-binding of membrane-associated valinomycin, a proton nuclear magnetic resonance study.

PubMed

Meers, P; Feigenson, G W

1988-03-03

Valinomycin, incorporated in small unilamellar vesicles of perdeuterated dimyristoylphosphatidylcholine, reveals several well-resolved 1H-NMR resonances. These resonances were used to examine the location, orientation and ion-binding of membrane-bound valinomycin. The order of affinity of membrane-bound valinomycin for cations is Rb+ greater than K+ greater than Cs+ greater than Ba2+, and binding is sensitive to surface change. The exchange between bound and free forms is fast on the NMR time scale. The intrinsic binding constants, extrapolated to zero anion concentration, are similar to those determined in aqueous solution. Rb+ and K+ show 1:1 binding to valinomycin, whereas the stoichiometry of Cs+ and Ba2+ is not certain. Paramagnetic chemical shift reagents and nitroxide spin label relaxation probes were used to study the location and orientation of valinomycin in the membrane. Despite relatively fast exchange of bound cations, the time average location of the cation-free form of valinomycin is deep within the bilayer under the conditions of these experiments. Upon complexation to K+, valinomycin moves closer to the interfacial region.

Single-molecule height measurements on microsomal cytochrome P450 in nanometer-scale phospholipid bilayer disks

NASA Astrophysics Data System (ADS)

Bayburt, Timothy H.; Sligar, Stephen G.

2002-05-01

The architecture of membrane proteins in their native environment of the phospholipid bilayer is critical for understanding physiological function, but has been difficult to realize experimentally. In this communication we describe the incorporation of a membrane-anchored protein into a supported phospholipid bilayer. Cytochrome P450 2B4 solubilized and purified from the hepatic endoplasmic reticulum was incorporated into phospholipid bilayer nanostructures and oriented on a surface for visualization by atomic force microscopy. Individual P450 molecules were observed protruding from the bilayer surface. Problems associated with deformation of the protein by the atomic force microscopy probe were avoided by analyzing force-dependent height measurements to quantitate the height of the protein above the bilayer surface. Measurements of the atomic force microscopy cantilever deflection as a function of probe-sample separation reveal that the top of the P450 opposite the N-terminal membrane anchor region sits 3.5 nanometers above the phospholipid-water boundary. Models of the orientation of the enzyme are presented and discussed in relation to membrane interactions and interaction with cytochrome P450 reductase.

Adrenocorticotropic Hormone (ACTH) Responses Require Actions of the Melanocortin-2 Receptor Accessory Protein on the Extracellular Surface of the Plasma Membrane.

PubMed

Malik, Sundeep; Dolan, Terrance M; Maben, Zachary J; Hinkle, Patricia M

2015-11-13

The melanocortin-2 (MC2) receptor is a G protein-coupled receptor that mediates responses to ACTH. The MC2 receptor acts in concert with the MC2 receptor accessory protein (MRAP) that is absolutely required for ACTH binding and signaling. MRAP has a single transmembrane domain and forms a highly unusual antiparallel homodimer that is stably associated with MC2 receptors at the plasma membrane. Despite the physiological importance of the interaction between the MC2 receptor and MRAP, there is little understanding of how the accessory protein works. The dual topology of MRAP has made it impossible to determine whether highly conserved and necessary regions of MRAP are required on the intracellular or extracellular face of the plasma membrane. The strategy used here was to fix the orientation of two antiparallel MRAP molecules and then introduce inactivating mutations on one side of the membrane or the other. This was achieved by engineering proteins containing tandem copies of MRAP fused to the amino terminus of the MC2 receptor. The data firmly establish that only the extracellular amino terminus (Nout) copy of MRAP, oriented with critical segments on the extracellular side of the membrane, is essential. The transmembrane domain of MRAP is also required in only the Nout orientation. Finally, activity of MRAP-MRAP-MC2-receptor fusion proteins with inactivating mutations in either MRAP or the receptor was rescued by co-expression of free wild-type MRAP or free wild-type receptor. These results show that the basic MRAP-MRAP-receptor signaling unit forms higher order complexes and that these multimers signal. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

Atomic resolution view into the structure–function relationships of the human myelin peripheral membrane protein P2

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

Ruskamo, Salla; University of Oulu, Oulu; Yadav, Ravi P.

2014-01-01

The structure of the human myelin peripheral membrane protein P2 has been refined at 0.93 Å resolution. In combination with functional experiments in vitro, in vivo and in silico, the fine details of the structure–function relationships in P2 are emerging. P2 is a fatty acid-binding protein expressed in vertebrate peripheral nerve myelin, where it may function in bilayer stacking and lipid transport. P2 binds to phospholipid membranes through its positively charged surface and a hydrophobic tip, and accommodates fatty acids inside its barrel structure. The structure of human P2 refined at the ultrahigh resolution of 0.93 Å allows detailed structuralmore » analyses, including the full organization of an internal hydrogen-bonding network. The orientation of the bound fatty-acid carboxyl group is linked to the protonation states of two coordinating arginine residues. An anion-binding site in the portal region is suggested to be relevant for membrane interactions and conformational changes. When bound to membrane multilayers, P2 has a preferred orientation and is stabilized, and the repeat distance indicates a single layer of P2 between membranes. Simulations show the formation of a double bilayer in the presence of P2, and in cultured cells wild-type P2 induces membrane-domain formation. Here, the most accurate structural and functional view to date on P2, a major component of peripheral nerve myelin, is presented, showing how it can interact with two membranes simultaneously while going through conformational changes at its portal region enabling ligand transfer.« less

The Pleckstrin Homology Domain of Phospholipase Cβ Transmits Enzymatic Activation through Modulation of Membrane - Domain Orientation§

PubMed Central

Drin, Guillaume; Douguet, Dominique; Scarlata, Suzanne

2008-01-01

Phospholipase C-beta (PLCβ) enzymes are activated by Gαq and Gβγ subunits and catalyze the hydrolysis of the minor membrane lipid phosphatidylinositol 4,5 bisphosphate (PI(4,5)P2). Activation of PLCβ2 by Gβγ subunits has been shown to be conferred through its N-terminal pleckstrin homology (PH) domain although the underlying mechanism is unclear. Also unclear are observations that the extent of Gβγ activation differs on different membrane surfaces. In this study, we have identified a unique region of the PH domain of PLCβ2 domain (residues 71-88) which, when added to the enzyme as a peptide, causes enzyme activation similar to Gβγ subunits. This PH domain segment interacts strongly with membranes composed of lipid mixtures but not those containing lipids with electrically neutral zwitterionic head groups. Moreso, addition of this segment perturbs interaction of the catalytic domain, but not the PH domain, with membrane surfaces. We monitored the orientation of the PH and catalytic domains of PLC by intermolecular fluorescence resonance energy transfer (FRET) using the Gβγ activatable mutant, PLCβ2/δ1(C193S). We find an increase in FRET with binding to membranes with mixed lipids but not to those containing only lipids with electrically neutral head groups. These results suggest that enzymatic activation can be conferred through optimal association of the PHβ71-88 region to specific membrane surfaces. These studies allow us to understand the basis of variations of Gβγ activation on different membrane surfaces. PMID:16669615

Influences of acid-base property of membrane on interfacial interactions related with membrane fouling in a membrane bioreactor based on thermodynamic assessment.

PubMed

Zhao, Leihong; Qu, Xiaolu; Zhang, Meijia; Lin, Hongjun; Zhou, Xiaoling; Liao, Bao-Qiang; Mei, Rongwu; Hong, Huachang

2016-08-01

Failure of membrane hydrophobicity in predicting membrane fouling requires a more reliable indicator. In this study, influences of membrane acid base (AB) property on interfacial interactions in two different interaction scenarios in a submerged membrane bioreactor (MBR) were studied according to thermodynamic approaches. It was found that both the polyvinylidene fluoride (PVDF) membrane and foulant samples in the MBR had relatively high electron donor (γ(-)) component and low electron acceptor (γ(+)) component. For both of interaction scenarios, AB interaction was the major component of the total interaction. The results showed that, the total interaction monotonically decreased with membrane γ(-), while was marginally affected by membrane γ(+), suggesting that γ(-) could act as a reliable indicator for membrane fouling prediction. This study suggested that membrane modification for fouling mitigation should orient to improving membrane surface γ(-) component rather than hydrophilicity. Copyright © 2016 Elsevier Ltd. All rights reserved.

On the alignment of cellulose microfibrils by cortical microtubules: a review and a model.

PubMed

Baskin, T I

2001-01-01

The hypothesis that microtubules align microfibrils, termed the alignment hypothesis, states that there is a causal link between the orientation of cortical microtubules and the orientation of nascent microfibrils. I have assessed the generality of this hypothesis by reviewing what is known about the relation between microtubules and microfibrils in a wide group of examples: in algae of the family Characeae, Closterium acerosum, Oocystis solitaria, and certain genera of green coenocytes and in land plant tip-growing cells, xylem, diffusely growing cells, and protoplasts. The salient features about microfibril alignment to emerge are as follows. Cellulose microfibrils can be aligned by cortical microtubules, thus supporting the alignment hypothesis. Alignment of microfibrils can occur independently of microtubules, showing that an alternative to the alignment hypothesis must exist. Microfibril organization is often random, suggesting that self-assembly is insufficient. Microfibril organization differs on different faces of the same cell, suggesting that microfibrils are aligned locally, not with respect to the entire cell. Nascent microfibrils appear to associate tightly with the plasma membrane. To account for these observations, I present a model that posits alignment to be mediated through binding the nascent microfibril. The model, termed templated incorporation, postulates that the nascent microfibril is incorporated into the cell wall by binding to a scaffold that is oriented; further, the scaffold is built and oriented around either already incorporated microfibrils or plasma membrane proteins, or both. The role of cortical microtubules is to bind and orient components of the scaffold at the plasma membrane. In this way, spatial information to align the microfibrils may come from either the cell wall or the cell interior, and microfibril alignment with and without microtubules are subsets of a single mechanism.

The research about microscopic structure of emulsion membrane in O/W emulsion by NMR and its influence to emulsion stability.

PubMed

Xie, Yiqiao; Chen, Jisheng; Zhang, Shu; Fan, Kaiyan; Chen, Gang; Zhuang, Zerong; Zeng, Mingying; Chen, De; Lu, Longgui; Yang, Linlin; Yang, Fan

2016-03-16

This paper discussed the influence of microstructure of emulsion membrane on O/W emulsion stability. O/W emulsions were emulsified with equal dosage of egg yolk lecithin and increasing dosage of co-emulsifier (oleic acid or HS15). The average particle size and centrifugal stability constant of emulsion, as well as interfacial tension between oil and water phase were determined. The microstructure of emulsion membrane had been studied by (1)H/(13)C NMR, meanwhile the emulsion droplets were visually presented with TEM and IFM. With increasing dosage of co-emulsifier, emulsions showed two stable states, under which the signal intensity of characteristic group (orient to lipophilic core) of egg yolk lecithin disappeared in NMR of emulsions, but that (orient to aqueous phase) of co-emulsifiers only had some reduction at the second stable state. At the two stable states, the emulsion membranes were neater in TEM and emulsion droplets were rounder in IFM. Furthermore, the average particle size of emulsions at the second stable state was bigger than that at the first stable state. Egg yolk lecithin and co-emulsifier respectively arranged into monolayer and bilayer emulsion membrane at the two stable states. The microstructure of emulsion membrane was related to the stability of emulsion. Copyright © 2016 Elsevier B.V. All rights reserved.

Fourier Transform Infrared and Resonance Raman Spectroscopic Studies of Bacteriorhodopsin.

NASA Astrophysics Data System (ADS)

Earnest, Thomas Nixon

Fourier transform infrared and resonance Raman spectroscopy were used to investigate the structure and function of the light-activated, transmembrane proton pump, bacteriorhodopsin, from the purple membrane of Halobacterium halobium. Bacteriorhodopsin (bR) is a 27,000 dalton integral membrane protein consisting of 248 amino acids with a retinylidene chromophore. Absorption of a photon leads to the translocation of one or two protons from the inside of the cell to the outside. Resonance Raman spectroscopy allows for the study of the configuration of retinal in bR and its photointermediates by the selective enhancement of vibrational modes of the chromophore. This technique was used to determine that the chromophore is attached to lysine-216 in both the bR _{570} and the M _{412} intermediates. In bR with tyrosine-64 selectively nitrated or aminated, the chromophore appears to have the same configuration in that bR _{570} (all- trans) and M _{412} (13- cis) states as it does in unmodified bR. Polarized Fourier transform infrared spectroscopy (FTIR) permits the study of the direction of transition dipole moments arising from molecular vibrations of the protein and the retinal chromophore. The orientation of alpha helical and beta sheet components was determined for bR with the average helical tilt found to lie mostly parallel to the membrane normal. The beta sheet structures also exhibit an IR linear dichroism for the amide I and amide II bands which suggest that the peptide backbone is mostly perpendicular to the membrane plane although it is difficult to determine whether the bands originate from sheet or turn components. The orientation of secondary structure components of the C-1 (residues 72-248) and C-2 (residues 1-71) fragments were also investigated to determine the structure of these putative membrane protein folding intermediates. Polarized, low temperature FTIR -difference spectroscopy was then used to investigate the structure of bR as it undergoes phototransitions from the light-adapted state, bR_{570} , to the K_{630} and M_{412} intermediates. The linear dichroism of C=C and C-C stretching modes, and the hydrogen out-of-plane (HOOP) modes of the chromophore show that the long axis of the polyene is 20-25^ circ out of the membrane plane and that the polyene plane is oriented mostly perpendicular to the membrane plane. Transition dipole moments from protein components are also investigated to determine the orientation of protein groups which undergo changes during the photocycle.

High resolution immunoelectron microscopic localization of functional domains of laminin, nidogen, and heparan sulfate proteoglycan in epithelial basement membrane of mouse cornea reveals different topological orientations.

PubMed

Schittny, J C; Timpl, R; Engel, J

1988-10-01

Thin and ultrathin cryosections of mouse cornea were labeled with affinity-purified antibodies directed against either laminin, its central segments (domain 1), the end of its long arm (domain 3), the end of one of its short arms (domain 4), nidogen, or low density heparan sulfate proteoglycan. All basement membrane proteins are detected by indirect immunofluorescence exclusively in the epithelial basement membrane, in Descemet's membrane, and in small amorphous plaques located in the stroma. Immunoelectron microscopy using the protein A-gold technique demonstrated laminin domain 1 and nidogen in a narrow segment of the lamina densa at the junction to the lamina lucida within the epithelial basement membrane. Domain 3 shows three preferred locations at both the cellular and stromal boundaries of the epithelial basement membrane and in its center. Domain 4 is located predominantly in the lamina lucida and the adjacent half of the lamina densa. The low density heparan sulfate proteoglycan is found all across the basement membrane showing a similar uniform distribution as with antibodies against the whole laminin molecule. In Descemet's membrane an even distribution was found with all these antibodies. It is concluded that within the epithelial basement membrane the center of the laminin molecule is located near the lamina densa/lamina lucida junction and that its long arm favors three major orientations. One is close to the cell surface indicating binding to a cell receptor, while the other two are directed to internal matrix structures. The apparent codistribution of laminin domain 1 and nidogen agrees with biochemical evidence that nidogen binds to this domain.

Efficient molecular mechanics simulations of the folding, orientation, and assembly of peptides in lipid bilayers using an implicit atomic solvation model

NASA Astrophysics Data System (ADS)

Bordner, Andrew J.; Zorman, Barry; Abagyan, Ruben

2011-10-01

Membrane proteins comprise a significant fraction of the proteomes of sequenced organisms and are the targets of approximately half of marketed drugs. However, in spite of their prevalence and biomedical importance, relatively few experimental structures are available due to technical challenges. Computational simulations can potentially address this deficit by providing structural models of membrane proteins. Solvation within the spatially heterogeneous membrane/solvent environment provides a major component of the energetics driving protein folding and association within the membrane. We have developed an implicit solvation model for membranes that is both computationally efficient and accurate enough to enable molecular mechanics predictions for the folding and association of peptides within the membrane. We derived the new atomic solvation model parameters using an unbiased fitting procedure to experimental data and have applied it to diverse problems in order to test its accuracy and to gain insight into membrane protein folding. First, we predicted the positions and orientations of peptides and complexes within the lipid bilayer and compared the simulation results with solid-state NMR structures. Additionally, we performed folding simulations for a series of host-guest peptides with varying propensities to form alpha helices in a hydrophobic environment and compared the structures with experimental measurements. We were also able to successfully predict the structures of amphipathic peptides as well as the structures for dimeric complexes of short hexapeptides that have experimentally characterized propensities to form beta sheets within the membrane. Finally, we compared calculated relative transfer energies with data from experiments measuring the effects of mutations on the free energies of translocon-mediated insertion of proteins into lipid bilayers and of combined folding and membrane insertion of a beta barrel protein.

Axis of Eye Rotation Changes with Head-Pitch Orientation during Head Impulses about Earth-Vertical

PubMed Central

Schubert, Michael C.; Clendaniel, Richard A.; Carey, John P.; Della Santina, Charles C.; Minor, Lloyd B.; Zee, David S.

2006-01-01

The goal of this study was to assess how the axis of head rotation, Listing's law, and eye position influence the axis of eye rotation during brief, rapid head rotations. We specifically asked how the axis of eye rotation during the initial angular vestibuloocular reflex (VOR) changed when the pitch orientation of the head relative to Earth-vertical was varied, but the initial position of the eye in the orbit and the orientation of Listing's plane with respect to the head were fixed. We measured three-dimensional eye and head rotation axes in eight normal humans using the search coil technique during head-and-trunk (whole-body) and head-on-trunk (head-only) “impulses” about an Earth-vertical axis. The head was initially oriented at one of five pitch angles (30° nose down, 15° nose down, 0°, 15° nose up, 30° nose up). The fixation target was always aligned with the nasooccipital axis. Whole-body impulses were passive, unpredictable, manual, rotations with peak-amplitude of ∼20°, peak-velocity of ∼80°/s, and peak-acceleration of ∼1000°/s2. Head-only impulses were also passive, unpredictable, manual, rotations with peak-amplitude of ∼20°, peak-velocity of ∼150°/s, and peak-acceleration of ∼3000°/s2. During whole-body impulses, the axis of eye rotation tilted in the same direction, and by an amount proportional (0.51 ± 0.09), to the starting pitch head orientation (P < 0.05). This proportionality constant decreased slightly to 0.39 ± 0.08 (P < 0.05) during head-only impulses. Using the head-only impulse data, with the head pitched up, we showed that only 50% of the tilt in the axis of eye rotation could be predicted from vectorial summation of the gains (eye velocity/head velocity) obtained for rotations about the pure yaw and roll head axes. Thus, even when the orientation of Listing's plane and eye position in the orbit are fixed, the axis of eye rotation during the VOR reflects a compromise between the requirements of Listing's law and a perfectly compensatory VOR. PMID:16552499

A Mach-Zehnder interferometer based on orbital angular momentum for improved vortex coronagraph efficiency

NASA Astrophysics Data System (ADS)

Piron, P.; Delacroix, C.; Huby, E.; Mawet, D.; Karlsson, M.; Ruane, G.; Habraken, S.; Absil, O.; Surdej, J.

2015-09-01

The Annular Groove Phase Mask (AGPM) is a vectorial vortex phase mask. It acts as a half-wave plate with a radial fast axis orientation operating in the mid infrared domain. When placed at the focus of a telescope element provides a continuous helical phase ramp for an on axis sources, which creates the orbital angular momentum. Thanks to that phase, the intensity of the central source is canceled by a down-stream pupil stop, while the off axis sources are not affected. However due to experimental conditions the nulling is hardly perfect. To improve the null, a Mach-Zehnder interferometer containing Dove prisms differently oriented can be proposed to sort out light based on its orbital angular momentum (OAM). Thanks to the differential rotation of the beam, a π phase shift is achieved for the on axis light affected by a non zero OAM. Therefore the contrast between the star and its faint companion is enhanced. Nevertheless, due the Dove prisms birefringence, the performance of the interferometer is relatively poor. To solve this problem, we propose to add a birefringent wave-plate in each arm to compensate this birefringence. In this paper, we will develop the mathematical model of the wave front using the Jones formalism. The performance of the interferometer is at first computed for the simple version without the birefringent plate. Then the effect of the birefringent plate is be mathematically described and the performance is re-computed.

A submerged membrane bioreactor with pendulum type oscillation (PTO) for oily wastewater treatment: membrane permeability and fouling control.

PubMed

Qin, Lei; Fan, Zheng; Xu, Lusheng; Zhang, Guoliang; Wang, Guanghui; Wu, Dexin; Long, Xuwei; Meng, Qin

2015-05-01

In this study, a novel submerged membrane bioreactor (SMBR) with pendulum type oscillation (PTO) hollow fiber membrane modules was developed to treat oily wastewater and control the problem of membrane fouling. To assess the potential of PTO membrane modules, the effect of oscillation orientation and frequency on membrane permeability was investigated in detail. The forces exerted on sludge flocs in the oscillating SMBR were analyzed to evaluate the impact of membrane oscillating on the cake layer resistance reduction. Results showed that the optimized PTO SMBR system exhibited 11 times higher membrane permeability and better fouling controllability than the conventional MBR system. By hydrodynamic analysis, it was found that the cooperative effect of bubble-induced turbulence and membrane oscillation in PTO SMBR system generated strong shear stress at liquid-membrane interface in vertical and horizontal direction and effectively hindered the particles from depositing on membrane surface. Copyright © 2015 Elsevier Ltd. All rights reserved.

An intracellular analysis of the visual responses of neurones in cat visual cortex.

PubMed Central

Douglas, R J; Martin, K A; Whitteridge, D

1991-01-01

1. Extracellular and intracellular recordings were made from neurones in the visual cortex of the cat in order to compare the subthreshold membrane potentials, reflecting the input to the neurone, with the output from the neurone seen as action potentials. 2. Moving bars and edges, generated under computer control, were used to stimulate the neurones. The membrane potential was digitized and averaged for a number of trials after stripping the action potentials. Comparison of extracellular and intracellular discharge patterns indicated that the intracellular impalement did not alter the neurones' properties. Input resistance of the neurone altered little during stable intracellular recordings (30 min-2 h 50 min). 3. Intracellular recordings showed two distinct patterns of membrane potential changes during optimal visual stimulation. The patterns corresponded closely to the division of S-type (simple) and C-type (complex) receptive fields. Simple cells had a complex pattern of membrane potential fluctuations, involving depolarizations alternating with hyperpolarizations. Complex cells had a simple single sustained plateau of depolarization that was often followed but not preceded by a hyperpolarization. In both simple and complex cells the depolarizations led to action potential discharges. The hyperpolarizations were associated with inhibition of action potential discharge. 4. Stimulating simple cells with non-optimal directions of motion produced little or no hyperpolarization of the membrane in most cases, despite a lack of action potential output. Directional complex cells always produced a single plateau of depolarization leading to action potential discharge in both the optimal and non-optimal directions of motion. The directionality could not be predicted on the basis of the position of the hyperpolarizing inhibitory potentials found in the optimal direction. 5. Stimulation of simple cells with non-optimal orientations occasionally produced slight hyperpolarizations and inhibition of action potential discharge. Complex cells, which had broader orientation tuning than simple cells, could show marked hyperpolarization for non-optimal orientations, but this was not generally the case. 6. The data do not support models of directionality and orientation that rely solely on strong inhibitory mechanisms to produce stimulus selectivity. PMID:1804981

A malaria model tested in the African savannah*

PubMed Central

Dietz, K.; Molineaux, L.; Thomas, A.

1974-01-01

A new mathematical model of malaria has been developed for comparing the effects of alternative control measures. It describes both the temporal changes of the P. falciparum infection rate and the immunity level of the population as a function of the dynamics and characteristics of the vector populations, which are summarized in the concept of vectorial capacity. A critical vectorial capacity is specified, below which malaria cannot maintain itself at an endemic level. The model has been tested with epidemiological data collected in a WHO research project in the African Savannah, Kano State, Northern Nigeria, since October 1970. The estimates of the model parameters were obtained by minimizing the χ2 function that measures the discrepancy between the observed and expected age-specific parasite rates in the two villages with the highest and the lowest vectorial capacity, respectively, at five surveys during one year of baseline data collection and between the observed and expected infant inoculation rates, in the main transmission seasons, in the same two villages. The model describes three aspects of immunity: loss of infectivity, loss of detectability, and increase of recovery rate. It is assumed that loss of infectivity precedes loss of detectability and increase of recovery rate. Superinfections are slowing down the recovery for high inoculation rates but do not reduce them to zero. They do not increase infectivity. PMID:4613512

Life cycle and vectorial competence of Triatoma williami (Galvão, Souza e Lima, 1965) under the influence of different blood meal sources.

PubMed

Lunardi, Rosaline Rocha; Gomes, Letícia Pinho; Peres Câmara, Thaís; Arrais-Silva, Wagner Welber

2015-09-01

Triatoma williami is naturally infected by Trypanosoma cruzi, the ethiological agent of Chagas disease, the most significant cause of morbidity and mortality in South and Central America.The possibility of domiciliation of T. williami increases the risk of human T. cruzi vetorial transmission. Despite this, there is a lack of data demonstrating the bionomic aspects, the vectorial competence or the natural ecotope and the wild hosts of T. williami. This study describes for the first time the life cycle of T. williami under the influence of two blood meal sources and also evaluates the vectorial potential of the species. The development of two groups of hundred triatomines was followed over the nymphal stages and adulthood. Each group was exposed to a sole blood meal source, mammalian or bird. The average egg-to-adult development time in both groups was similar, except by shorter stages of N3 and N4 in triatomines fed on mammals. The group fed on birds needed more blood feedings to suffer the ecdysis and had higher cumulative mortality in the nymphal stages. Although the observed delay at defecation of adults after feeding, our results suggest that T. williami in the third and fifth nymphal stages may be good vectors. Copyright © 2015 Elsevier B.V. All rights reserved.

Vectorial capacity and vector control: reconsidering sensitivity to parameters for malaria elimination

PubMed Central

Brady, Oliver J.; Godfray, H. Charles J.; Tatem, Andrew J.; Gething, Peter W.; Cohen, Justin M.; McKenzie, F. Ellis; Perkins, T. Alex; Reiner, Robert C.; Tusting, Lucy S.; Sinka, Marianne E.; Moyes, Catherine L.; Eckhoff, Philip A.; Scott, Thomas W.; Lindsay, Steven W.; Hay, Simon I.; Smith, David L.

2016-01-01

Background Major gains have been made in reducing malaria transmission in many parts of the world, principally by scaling-up coverage with long-lasting insecticidal nets and indoor residual spraying. Historically, choice of vector control intervention has been largely guided by a parameter sensitivity analysis of George Macdonald's theory of vectorial capacity that suggested prioritizing methods that kill adult mosquitoes. While this advice has been highly successful for transmission suppression, there is a need to revisit these arguments as policymakers in certain areas consider which combinations of interventions are required to eliminate malaria. Methods and Results Using analytical solutions to updated equations for vectorial capacity we build on previous work to show that, while adult killing methods can be highly effective under many circumstances, other vector control methods are frequently required to fill effective coverage gaps. These can arise due to pre-existing or developing mosquito physiological and behavioral refractoriness but also due to additive changes in the relative importance of different vector species for transmission. Furthermore, the optimal combination of interventions will depend on the operational constraints and costs associated with reaching high coverage levels with each intervention. Conclusions Reaching specific policy goals, such as elimination, in defined contexts requires increasingly non-generic advice from modelling. Our results emphasize the importance of measuring baseline epidemiology, intervention coverage, vector ecology and program operational constraints in predicting expected outcomes with different combinations of interventions. PMID:26822603

Demonstration of a vectorial optical field generator with adaptive close loop control.

PubMed

Chen, Jian; Kong, Lingjiang; Zhan, Qiwen

2017-12-01

We experimentally demonstrate a vectorial optical field generator (VOF-Gen) with an adaptive close loop control. The close loop control capability is illustrated with the calibration of polarization modulation of the system. To calibrate the polarization ratio modulation, we generate 45° linearly polarized beam and make it propagate through a linear analyzer whose transmission axis is orthogonal to the incident beam. For the retardation calibration, circularly polarized beam is employed and a circular polarization analyzer with the opposite chirality is placed in front of the CCD as the detector. In both cases, the close loop control automatically changes the value of the corresponding calibration parameters in the pre-set ranges to generate the phase patterns applied to the spatial light modulators and records the intensity distribution of the output beam by the CCD camera. The optimized calibration parameters are determined corresponding to the minimum total intensity in each case. Several typical kinds of vectorial optical beams are created with and without the obtained calibration parameters, and the full Stokes parameter measurements are carried out to quantitatively analyze the polarization distribution of the generated beams. The comparisons among these results clearly show that the obtained calibration parameters could remarkably improve the accuracy of the polarization modulation of the VOF-Gen, especially for generating elliptically polarized beam with large ellipticity, indicating the significance of the presented close loop in enhancing the performance of the VOF-Gen.

Graphene can wreak havoc with cell membranes.

PubMed

Dallavalle, Marco; Calvaresi, Matteo; Bottoni, Andrea; Melle-Franco, Manuel; Zerbetto, Francesco

2015-02-25

Molecular dynamics--coarse grained to the level of hydrophobic and hydrophilic interactions--shows that small hydrophobic graphene sheets pierce through the phospholipid membrane and navigate the double layer, intermediate size sheets pierce the membrane only if a suitable geometric orientation is met, and larger sheets lie mainly flat on the top of the bilayer where they wreak havoc with the membrane and create a patch of upturned phospholipids. The effect arises in order to maximize the interaction between hydrophobic moieties and is quantitatively explained in terms of flip-flops by the analysis of the simulations. Possible severe biological consequences are discussed.

Impact of cholesterol on voids in phospholipid membranes

NASA Astrophysics Data System (ADS)

Falck, Emma; Patra, Michael; Karttunen, Mikko; Hyvönen, Marja T.; Vattulainen, Ilpo

2004-12-01

Free volume pockets or voids are important to many biological processes in cell membranes. Free volume fluctuations are a prerequisite for diffusion of lipids and other macromolecules in lipid bilayers. Permeation of small solutes across a membrane, as well as diffusion of solutes in the membrane interior are further examples of phenomena where voids and their properties play a central role. Cholesterol has been suggested to change the structure and function of membranes by altering their free volume properties. We study the effect of cholesterol on the properties of voids in dipalmitoylphosphatidylcholine (DPPC) bilayers by means of atomistic molecular dynamics simulations. We find that an increasing cholesterol concentration reduces the total amount of free volume in a bilayer. The effect of cholesterol on individual voids is most prominent in the region where the steroid ring structures of cholesterol molecules are located. Here a growing cholesterol content reduces the number of voids, completely removing voids of the size of a cholesterol molecule. The voids also become more elongated. The broad orientational distribution of voids observed in pure DPPC is, with a 30% molar concentration of cholesterol, replaced by a distribution where orientation along the bilayer normal is favored. Our results suggest that instead of being uniformly distributed to the whole bilayer, these effects are localized to the close vicinity of cholesterol molecules.

Imaging plasma membrane deformations with pTIRFM.

PubMed

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

2014-04-02

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

[The role of the Aedes aegypti vector in the epidemiology of dengue in Mexico].

PubMed

Fernández-Salas, I; Flores-Leal, A

1995-01-01

The role of Aedes aegypti (Lineo) in the epidemiology of dengue fever in Mexico is herein discussed based on the vectorial capacity model. Comments on the advantages and disadvantages of each model component at the time of field determinations are also presented. Emphasis is made on the impact of sampling and method bias on the results of vectorial capacity studies. The paper also addresses the need to increase vector biology knowledge as an input for epidemiological work to explain and predict dengue fever outbreaks. Comments on potential entomological variables not considered by the quantitative model are included. Finally, we elaborate on the introduction of Aedes albopictus (Skuse) in Mexico as a new risk factor and on its implications for the understanding of dengue fever transmission in Mexico.

Orally-transmitted Chagas disease.

PubMed

Filigheddu, Maria Teresa; Górgolas, Miguel; Ramos, José Manuel

2017-02-09

Chagas disease is a zoonosis caused by protozoan parasite Trypanosoma cruzi, which is most frequently associated with a vectorial transmission. However, in recent years we have observed a significant increase in the oral transmission of the disease, associated mainly with the consumption of drinks made from fruit or other vegetables contaminated with triatomine faeces or secretions from infected mammals. After a latency period of 3 to 22 days after ingestion, the oral infection is characterized by more severe manifestations than those associated with vectorial transmission: prolonged fever, acute myocarditis with heart failure and, in some cases, meningoencephalitis. Mortality can reach up to 33% of those infected. The aim of this paper is to review this matter and to promote prevention practices. Copyright © 2016 Elsevier España, S.L.U. All rights reserved.

Reflection type metasurface designed for high efficiency vectorial field generation

NASA Astrophysics Data System (ADS)

Wang, Shiyi; Zhan, Qiwen

2016-07-01

We propose a reflection type metal-insulator-metal (MIM) metasurface composed of hybrid nano-antennas for comprehensive spatial engineering of the properties of optical fields. The capability of such structure is illustrated in the design of a device that can be used to produce a radially polarized vectorial beam for optical needle field generation. This device consists of uniformly segmented sectors of high efficiency MIM metasurface. With each of the segment sector functioning as a local quarter-wave-plate (QWP), the device is designed to convert circularly polarized incidence into local linear polarization to create an overall radial polarization with corresponding binary phases and extremely high dynamic range amplitude modulation. The capability of such devices enables the generation of nearly arbitrarily complex optical fields that may find broad applications that transcend disciplinary boundaries.

Scanning metallic nanosphere microscopy for vectorial profiling of optical focal spots.

PubMed

Yi, Hui; Long, Jing; Li, Hongquan; He, Xiaolong; Yang, Tian

2015-04-06

Recent years have witnessed fast progress in the development of spatially variant states of polarization under high numerical aperture focusing, and intensive exploration of their applications. We report a vectorial, broadband, high contrast and subwavelength resolution method for focal spot profiling. In this experiment, a 100 nm diameter gold nanosphere on a silica aerogel substrate is raster scanned across the focal spots, and the orthogonal polarization components can be obtained simultaneously by measuring the scattering far field in a confocal manner. The metallic-nanosphere-on-aerogel structure ensures negligible distortion to the focal spots, low crosstalk between orthogonal polarization components (1/39 in experiment), and a low level background noise (1/80 of peak intensity in experiment), while high contrast imaging is not limited by the resonance bandwidth.

Single Particle Orientation and Rotational Tracking (SPORT) in biophysical studies

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

Gu, Yan; Ha, Ji Won; Augspurger, Ashley E.

The single particle orientation and rotational tracking (SPORT) techniques have seen rapid development in the past 5 years. Recent technical advances have greatly expanded the applicability of SPORT in biophysical studies. In this feature article, we survey the current development of SPORT and discuss its potential applications in biophysics, including cellular membrane processes and intracellular transport.

The establishment of polarized membrane traffic in Xenopus laevis embryos.

PubMed

Roberts, S J; Leaf, D S; Moore, H P; Gerhart, J C

1992-09-01

Delineation of apical and basolateral membrane domains is a critical step in the epithelialization of the outer layer of cells in the embryo. We have examined the initiation of polarized membrane traffic in Xenopus and show that membrane traffic is not polarized in oocytes but polarized membrane domains appear at first cleavage. The following proteins encoded by injected RNA transcripts were used as markers to monitor membrane traffic: (a) VSV G, a transmembrane glycoprotein preferentially inserted into the basolateral surface of polarized epithelial cells; (b) GThy-1, a fusion protein of VSV G and Thy-1 that is localized to the apical domains of polarized epithelial cells; and (c) prolactin, a peptide hormone that is not polarly secreted. In immature oocytes, there is no polarity in the expression of VSV G or GThy-1, as shown by the constitutive expression of both proteins at the surface in the animal and vegetal hemispheres. At meiotic maturation, membrane traffic to the surface is blocked; the plasma membrane no longer accepts the vesicles synthesized by the oocyte (Leaf, D. L., S. J. Roberts, J. C. Gerhart, and H.-P. Moore. 1990. Dev. Biol. 141:1-12). When RNA transcripts are injected after fertilization, VSV G is expressed only in the internal cleavage membranes (basolateral orientation) and is excluded from the outer surface (apical orientation, original oocyte membrane). In contrast, GThy-1 and prolactin, when expressed in embryos, are inserted or released at both the outer membrane derived from the oocyte and the inner cleavage membranes. Furthermore, not all of the cleavage membrane comes from an embryonic pool of vesicles--some of the cleavage membrane comes from vesicles synthesized during oogenesis. Using prolactin as a marker, we found that a subset of vesicles synthesized during oogenesis was only released after fertilization. However, while embryonic prolactin was secreted from both apical and basolateral surfaces, the secretion of oogenic prolactin was polarized. Oogenic prolactin was secreted only into the blastocoel (from the cleavage membrane), none could be detected in the external medium (from the original oocyte membrane). These results provide the first direct evidence that the oocyte synthesizes a cache of vesicles for specific recruitment to the embryonic cleavage membranes which are polarized beginning with the first cleavage division.

Sided functions of an arginine-agmatine antiporter oriented in liposomes.

PubMed

Tsai, Ming-Feng; Fang, Yiling; Miller, Christopher

2012-02-28

The arginine-dependent extreme acid resistance system helps enteric bacteria survive the harsh gastric environment. At the center of this multiprotein system is an arginine-agmatine antiporter, AdiC. To maintain cytoplasmic pH, AdiC imports arginine and exports its decarboxylated product, agmatine, resulting in a net extrusion of one "virtual proton" in each turnover. The random orientation of AdiC in reconstituted liposomes throws up an obstacle to quantifying its transport mechanism. To overcome this problem, we introduced a mutation, S26C, near the substrate-binding site. This mutant exhibits substrate recognition and pH-dependent activity similar to those of the wild-type protein but loses function completely upon reaction with thiol reagents. The membrane-impermeant MTSES reagent can then be used as a cleanly sided inhibitor to silence those S26C-AdiC proteins whose extracellular portion projects from the external side of the liposome. Alternatively, the membrane-permeant MTSEA and membrane-impermeant reducing reagent, TCEP, can be used together to inhibit proteins in the opposite orientation. This approach allows steady-state kinetic analysis of AdiC in a sided fashion. Arginine and agmatine have similar Michaelis-Menten parameters for both sides of the protein, while the extracellular side selects arginine over argininamide, a mimic of the carboxylate-protonated form of arginine, more effectively than does the cytoplasmic side. Moreover, the two sides of AdiC have different pH sensitivities. AdiC activity increases to a plateau at pH 4 as the extracellular side is acidified, while the cytoplasmic side shows an optimal pH of 5.5, with further acidification inhibiting transport. This oriented system allows more precise analysis of AdiC-mediated substrate transport than has been previously available and permits comparison to the situation experienced by the bacterial membrane under acid stress.

Structure and Orientation of Bovine Lactoferrampin in the Mimetic Bacterial Membrane as Revealed by Solid-State NMR and Molecular Dynamics Simulation

PubMed Central

Tsutsumi, Atsushi; Javkhlantugs, Namsrai; Kira, Atsushi; Umeyama, Masako; Kawamura, Izuru; Nishimura, Katsuyuki; Ueda, Kazuyoshi; Naito, Akira

2012-01-01

Bovine lactoferrampin (LFampinB) is a newly discovered antimicrobial peptide found in the N1-domain of bovine lactoferrin (268–284), and consists of 17 amino-acid residues. It is important to determine the orientation and structure of LFampinB in bacterial membranes to reveal the antimicrobial mechanism. We therefore performed 13C and 31P NMR, 13C-31P rotational echo double resonance (REDOR), potassium ion-selective electrode, and quartz-crystal microbalance measurements for LFampinB with mimetic bacterial membrane and molecular-dynamics simulation in acidic membrane. 31P NMR results indicated that LFampinB caused a defect in mimetic bacterial membranes. Ion-selective electrode measurements showed that ion leakage occurred for the mimetic bacterial membrane containing cardiolipin. Quartz-crystal microbalance measurements revealed that LFampinB had greater affinity to acidic phospholipids than that to neutral phospholipids. 13C DD-MAS and static NMR spectra showed that LFampinB formed an α-helix in the N-terminus region and tilted 45° to the bilayer normal. REDOR dephasing patterns between carbonyl carbon nucleus in LFampinB and phosphorus nuclei in lipid phosphate groups were measured by 13C-31P REDOR and the results revealed that LFampinB is located in the interfacial region of the membrane. Molecular-dynamics simulation showed the tilt angle to be 42° and the rotation angle to be 92.5° for Leu3, which are in excellent agreement with the experimental values. PMID:23083717

Micropaleontological and Paleomagnetic Characterization of La Ceiba K/T Boundary Section, Central Mexico

NASA Astrophysics Data System (ADS)

Martínez-López, M.; Urrutia-Fucugauchi, J.

2007-05-01

We report results of a micropaleontological and magnetostratigraphic study of the La Ceiba section that spans the K/T boundary. La Ceiba is located in central Mexico (20o 19.8' N, 97o 41.0' W) within the Tampico-Mizantla basin. The K/T boundary is marked by a clastic unit of about one-meter thickness intercalated between the carbonate hemipelagic marls of the Cretaceous Mendez Formation and the Paleocene Velasco Formation. The clastic unit can be divided into four sub-units, according to their texture and architectural characteristics following Arenillas et al. (2002). The basal sub-unit is formed by calcareous marls and is rich in shocked quartz and millimeter size spherules with microtektites and bioclasts of shallow water origin. The second sub-unit is formed by medium-grained sandstones, with clasts and quartz fragments, feldspars, metamorphic and calcareous fragments and re-worked benthic and planktic foraminifera. The third sub-unit is composed by a single body of medium- to fine-grained sandstones with tabular geometry. In this sub-unit, cross- and parallel-lamination trough cross-stratification, current ripples and climbing ripples have been observed. The top sub-unit is a tabular body of fine-grained sandstones, showing parallel-lamination and low-angle cross-lamination, with asymmetric ripples and burrow traces to the top. For the paleontologic and paleomagnetic study we collected twenty-five oriented samples across the section. We measured the low-field susceptibility, intensity and direction of the NRM. The vectorial composition and stability of NRM were analyzed by progressive thermal and alternating field demagnetization. Vectorial orthogonal diagrams and vector subtraction and principal component analysis were used to determine the characteristic magnetization and secondary components for each sample. The characteristic NRM negative inclination and southward declination in the K/T clastic sediments indicate a reverse polarity, which is correlated to reverse chron 29r that spans the K/T boundary. Micropaleontology analyses permit identification of six biozones. Two biozones (biozone of Rugoglobigerina scootti and Abathomphalus mayaroensis) correspond to the Maastrichtian. Four biozones (Guembelitria cretácea, Parvularogoglobigerina eugubina, Parasubotina pseudobulloides and Acarina trinidadensis) correspond to the Danian.

Asymmetric bi-layer PFSA membranes as model systems for the study of water management in the PEMFC.

PubMed

Peng, Z; Peng, A Z; Morin, A; Huguet, P; Lanteri, Y; Deabate, S

2014-10-14

New bi-layer PFSA membranes made of Nafion® NRE212 and Aquivion™ E79-05s with different equivalent weights are prepared with the aim of managing water repartition in the PEMFC. The membrane water transport properties, i.e. back-diffusion and electroosmosis, as well as the electrochemical performances, are compared to those of state-of-art materials. The actual water content (the inner water concentration profile across the membrane thickness) is measured under operation in the fuel cell by in situ Raman microspectroscopy. The orientation of the equivalent weight gradient with respect to the water external gradient and to the proton flow direction affects the membrane water content, the water transport ability and, thus, the fuel cell performances. Higher power outputs, related to lower ohmic losses, are observed when the membrane is assembled with the lower equivalent weight layer (Aquivion™) at the anode side. This orientation, corresponding to enhanced water transport by back-flow while electroosmosis remains unaffected, results in the higher hydration of the membrane and of the anode active layer during operation. Also, polarization data suggest a different water repartition in the fuel cell along the on-plane direction. Even if the interest in multi-layer PFSA membranes as perspective electrolytes for PEMFCs is not definitively attested, these materials appear to be excellent model systems to establish relationships between the membrane transport properties, the water distribution in the fuel cell and the electrochemical performances. Thanks to the micrometric resolution, in situ Raman microspectroscopy proves to be a unique tool to measure the actual hydration of the membrane at the surface swept by the hydrated feed gases during operation, so that it can be used as a local probe of the water concentration evolution along the gas distribution channels according to changing working conditions.

Effect of pretreatment and membrane orientation on fluxes for concentration of whey with high foulants by using NH3/CO2 in forward osmosis.

PubMed

Seker, M; Buyuksari, E; Topcu, S; Babaoglu, D S; Celebi, D; Keskinler, B; Aydiner, C

2017-11-01

Usage of forward osmosis membrane in FO mode, in which active and support layers of the membrane were in contact with the feed and the draw solutions respectively, provided higher initial water flux (12L/m 2 h) than the usage of membrane in PRO mode (6L/m 2 h) having opposite orientation but fluxes approached to each other after 4h during concentration of whey with NH 3 /CO 2 as draw salt. High organic and inorganic foulants of whey was considered as reason for observed result in addition to lower solute resistivity. Initial water flux (8,5L/m 2 h) was lower when pre-treatment was applied before forward osmosis process but final flux (4L/m 2 h) was equal flux of non pre-treatment. Reduction of solute resistivity or absence of hydraulic pressure can be reasons for lower initial flux. Detection of organic carbon but absence of lactose in draw solution showed passage of molecules being different than lactose into draw solution. Copyright © 2017 Elsevier Ltd. All rights reserved.

Nanoparticle orientation to control RNA loading and ligand display on extracellular vesicles for cancer regression

NASA Astrophysics Data System (ADS)

Pi, Fengmei; Binzel, Daniel W.; Lee, Tae Jin; Li, Zhefeng; Sun, Meiyan; Rychahou, Piotr; Li, Hui; Haque, Farzin; Wang, Shaoying; Croce, Carlo M.; Guo, Bin; Evers, B. Mark; Guo, Peixuan

2018-01-01

Nanotechnology offers many benefits, and here we report an advantage of applying RNA nanotechnology for directional control. The orientation of arrow-shaped RNA was altered to control ligand display on extracellular vesicle membranes for specific cell targeting, or to regulate intracellular trafficking of small interfering RNA (siRNA) or microRNA (miRNA). Placing membrane-anchoring cholesterol at the tail of the arrow results in display of RNA aptamer or folate on the outer surface of the extracellular vesicle. In contrast, placing the cholesterol at the arrowhead results in partial loading of RNA nanoparticles into the extracellular vesicles. Taking advantage of the RNA ligand for specific targeting and extracellular vesicles for efficient membrane fusion, the resulting ligand-displaying extracellular vesicles were capable of specific delivery of siRNA to cells, and efficiently blocked tumour growth in three cancer models. Extracellular vesicles displaying an aptamer that binds to prostate-specific membrane antigen, and loaded with survivin siRNA, inhibited prostate cancer xenograft. The same extracellular vesicle instead displaying epidermal growth-factor receptor aptamer inhibited orthotopic breast cancer models. Likewise, survivin siRNA-loaded and folate-displaying extracellular vesicles inhibited patient-derived colorectal cancer xenograft.

Determination of Structural Topology of a Membrane Protein in Lipid -Bilayers using Polarization Optimized Experiments (POE) for Static and MAS Solid State NMR Spectroscopy

PubMed Central

Mote, Kaustubh R.; Gopinath, T.; Veglia, Gianluigi

2013-01-01

The low sensitivity inherent to both the static and magic angle spinning techniques of solid-state NMR (ssNMR) spectroscopy has thus far limited the routine application of multidimensional experiments to determine the structure of membrane proteins in lipid bilayers. Here, we demonstrate the advantage of using a recently developed class of experiments, polarization optimized experiments (POE), for both static and MAS spectroscopy to achieve higher sensitivity and substantial time-savings for 2D and 3D experiments. We used sarcolipin, a single pass membrane protein, reconstituted in oriented bicelles (for oriented ssNMR) and multilamellar vesicles (for MAS ssNMR) as a benchmark. The restraints derived by these experiments are then combined into a hybrid energy function to allow simultaneous determination of structure and topology. The resulting structural ensemble converged to a helical conformation with a backbone RMSD ∼ 0.44 Å, a tilt angle of 24° ± 1°, and an azimuthal angle of 55° ± 6°. This work represents a crucial first step toward obtaining high-resolution structures of large membrane proteins using combined multidimensional O-ssNMR and MAS-ssNMR. PMID:23963722

A comparative study of different amniotic membrane orientations during extraocular muscle surgery in rabbits.

PubMed

Kassem, Rehab Rashad; El-Mofty, Randa Mohamed Abdel-Moneim; Khodeir, Mustafa Mahmoud; Hamza, Wael Mostafa

2018-03-01

To histopathologically compare the effect of different orientations of cryopreserved human amniotic membrane (AM) transplant during extraocular muscle surgery in rabbits. Fifty-two albino rabbit eyes underwent 4-mm resection of the superior rectus. Eyes were randomly divided into four groups. In Group C (Control group, 16 eyes) the muscle was not wrapped with amniotic membrane. In the three AM groups, cryopreserved AM was wrapped around the muscle, oriented with either its stroma (Group S, 15 eyes) or epithelium (Group E, nine eyes) towards the muscle, or folded on itself with the epithelium externally (Group F, 12 eyes). The rabbits were sacrificed and the eyes were enucleated 6 weeks after surgery. Histopathological examination was conducted for periamniotic, foreign body, scleral, and conjunctival inflammation, conjunctival vascularity, adhesions and muscle fibrosis. In all AM eyes, the AM was surrounded by periamniotic inflammation, with no adhesions detected between the muscle and surrounding tissues in the segment where the AM was present, but detected elsewhere. Adhesions were detected in all group C eyes. Foreign body inflammation was significantly less in Group C than in each of the AM groups (p < .05), but was insignificantly different among the three AM groups (p > .05). Scleral inflammation was absent in all specimens. No significant differences were noted among all groups in terms of conjunctival vascularity, conjunctival inflammation, or muscle fibrosis (p > .05). All AM orientations were equally effective in preventing the development of postoperative adhesions between the extraocular muscle and surrounding tissues.

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

NASA Astrophysics Data System (ADS)

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

2016-12-01

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

Mode and polarization state selected guided wave spectroscopy of orientational anisotrophy in model membrane cellulosic polymer films: relevance to lab-on-a-chip

NASA Astrophysics Data System (ADS)

Andrews, Mark P.; Kanigan, Tanya

2007-06-01

Orientation anisotropies in structural properties relevant to the use of cellulosic polymers as membranes for lab-on-chips were investigated for cellulose acetate (CA) and regenerated cellulose (RC) films deposited as slab waveguides. Anisotropy was probed with mode and polarization state selected guided wave Raman spectroscopy. CA exhibits partial chain orientation in the plane of the film, and this orientation is independent of sample substrate and film preparation conditions. RC films also show in-plane anisotropy, where the hexose sugar rings lie roughly in the plane of the film. Explanations are given of the role of artifacts in interpreting waveguide Raman spectra, including anomalous contributions to Raman spectra that arise from deviations from right angle scattering geometry, mode-dependent contributions to longitudinal electric field components and TE<-->TM mode conversion. We explore diffusion profiles of small molecules in cellulosic films by adaptations of an inverse-Wentzel-Kramers-Brillouin (iWKB) recursive, noninteger virtual mode index algorithm. Perturbations in the refractive index distribution, n(z), are recovered from the measured relative propagation constants, neffective,m, of the planar waveguide. The refractive index distribution then yields the diffusion profile.

Protein membrane interaction: effect of myelin basic protein on the dynamics of oriented lipids

NASA Astrophysics Data System (ADS)

Natali, F.; Relini, A.; Gliozzi, A.; Rolandi, R.; Cavatorta, P.; Deriu, A.; Fasano, A.; Riccio, P.

2003-08-01

We have studied the effect of physiological amounts of myelin basic protein (MBP) on pure dimyristoyl L-α-phosphatidic acid (DMPA) oriented membranes. The investigation has been carried out using several complementary experimental methods to provide a detailed characterization of the proteo-lipid complexes. In particular, taking advantage of the power of the quasi-elastic neutron scattering (QENS) technique as optimal probe in biology, a significant effect is suggested to be induced by MBP on the anisotropy of lipid dynamics across the liquid-gel phase transition. Thus, the enhancement of the spatially restricted, vertical translation motion of DMPA is suggested to be the main responsible for the increased contribution of the out of plane lipid dynamics observed at 340 K.

Active processes make mixed lipid membranes either flat or crumpled

NASA Astrophysics Data System (ADS)

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.

The Sodium Glucose Cotransporter SGLT1 Is an Extremely Efficient Facilitator of Passive Water Transport.

PubMed

Erokhova, Liudmila; Horner, Andreas; Ollinger, Nicole; Siligan, Christine; Pohl, Peter

2016-04-29

The small intestine is void of aquaporins adept at facilitating vectorial water transport, and yet it reabsorbs ∼8 liters of fluid daily. Implications of the sodium glucose cotransporter SGLT1 in either pumping water or passively channeling water contrast with its reported water transporting capacity, which lags behind that of aquaporin-1 by 3 orders of magnitude. Here we overexpressed SGLT1 in MDCK cell monolayers and reconstituted the purified transporter into proteoliposomes. We observed the rate of osmotic proteoliposome deflation by light scattering. Fluorescence correlation spectroscopy served to assess (i) SGLT1 abundance in both vesicles and plasma membranes and (ii) flow-mediated dilution of an aqueous dye adjacent to the cell monolayer. Calculation of the unitary water channel permeability, pf, yielded similar values for cell and proteoliposome experiments. Neither the absence of glucose or Na(+), nor the lack of membrane voltage in vesicles, nor the directionality of water flow grossly altered pf Such weak dependence on protein conformation indicates that a water-impermeable occluded state (glucose and Na(+) in their binding pockets) lasts for only a minor fraction of the transport cycle or, alternatively, that occlusion of the substrate does not render the transporter water-impermeable as was suggested by computational studies of the bacterial homologue vSGLT. Although the similarity between the pf values of SGLT1 and aquaporin-1 makes a transcellular pathway plausible, it renders water pumping physiologically negligible because the passive flux would be orders of magnitude larger. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

Beam shaping with vortex beam generated by liquid crystal spatial light modulator

NASA Astrophysics Data System (ADS)

Gao, Yue; Liu, Ke; Sun, Zeng-yu; Guo, Lei; Gan, Yu

2015-02-01

An optical vortex is a beam of light with phase varying in a corkscrew-like manner along its direction of propagation and so has a helical wavefront. When such a vectorial vortex beam and the Gaussian beam with orthogonal polarization are focused by low NA lens, the Gaussian component causes a focal intensity distribution with a solid center and the vortex component causes a donut distribution with hollow dark center. The shape of the focus can be continuously varied by continuously adjusting the relative weight of the two components. Flat top focusing can be obtained under appropriate conditions. It is demonstrated through experiments with a liquid crystal spatial light modulator in such a beam, that flattop focus can be obtained by vectorial vortex beams with topological charge of +1 to achieve beam shaping vortex.

Bedbugs and Infectious Diseases

PubMed Central

Blanc, Véronique; Del Giudice, Pascal; Levy-Bencheton, Anna; Chosidow, Olivier; Marty, Pierre; Brouqui, Philippe

2011-01-01

Bedbugs are brown and flat hematophagous insects. The 2 cosmopolite species, Cimex lectularius and Cimex hemipterus, feed on humans and/or domestic animals, and recent outbreaks have been reported in occidental countries. Site assessment for bedbug eradication is complex but can be assured, despite emerging insecticide resistance, by hiring a pest-control manager. The common dermatological presentation of bites is an itchy maculopapular wheal. Urticarial reactions and anaphylaxis can also occur. Bedbugs are suspected of transmitting infectious agents, but no report has yet demonstrated that they are infectious disease vectors. We describe 45 candidate pathogens potentially transmitted by bedbugs, according to their vectorial capacity, in the wild, and vectorial competence, in the laboratory. Because of increasing demands for information about effective control tactics and public health risks of bedbugs, continued research is needed to identify new pathogens in wild Cimex species (spp) and insecticide resistance. PMID:21288844

Full-vectorial finite element method in a cylindrical coordinate system for loss analysis of photonic wire bends

NASA Astrophysics Data System (ADS)

Kakihara, Kuniaki; Kono, Naoya; Saitoh, Kunimasa; Koshiba, Masanori

2006-11-01

This paper presents a new full-vectorial finite-element method in a local cylindrical coordinate system, to effectively analyze bending losses in photonic wires. The discretization is performed in the cross section of a three-dimensional curved waveguide, using hybrid edge/nodal elements. The solution region is truncated by anisotropic, perfectly matched layers in the cylindrical coordinate system, to deal properly with leaky modes of the waveguide. This approach is used to evaluate bending losses in silicon wire waveguides. The numerical results of the present approach are compared with results calculated with an equivalent straight waveguide approach and with reported experimental data. These comparisons together demonstrate the validity of the present approach based on the cylindrical coordinate system and also clarifies the limited validity of the equivalent straight waveguide approximation.

The vectorial control of magnetization by light.

PubMed

Kanda, Natsuki; Higuchi, Takuya; Shimizu, Hirokatsu; Konishi, Kuniaki; Yoshioka, Kosuke; Kuwata-Gonokami, Makoto

2011-06-21

Application of coherent light-matter interactions has recently been extended to the ultrafast control of magnetization. An important but unrealized technique is the manipulation of magnetization vector motion to make it follow an arbitrarily designed multidimensional trajectory. Here we demonstrate a full manipulation of two-dimensional magnetic oscillations in antiferromagnetic NiO with a pair of polarization-twisted femtosecond laser pulses. We employ Raman-type nonlinear optical processes, wherein magnetic oscillations are impulsively induced with a controlled initial phase. Their azimuthal angle follows well-defined selection rules that have been determined by the symmetries of the materials. We emphasize that the temporal variation of the laser-pulse polarization angle enables us to control the phase and amplitude of the two degenerate modes, independently. These results lead to a new concept of the vectorial control of magnetization by light.

Role of the Cotyledons in the Phototropic Response of Lavatera cretica Seedlings.

PubMed

Schwartz, A; Koller, D

1980-07-01

Young seedlings of Lavatera cretica L. exhibit positive phototropism. The hypocotyl perceives unilateral illumination with blue light and curves towards the light source by unequal growth. In addition, the cotyledonary laminas perceive the vectorial component of unilateral illumination with blue light and reorient normal to the beam by creating a turgor differential in their pulvini. Excision of one cotyledon resulted in negative organotropic curvature of the hypocotyl, away from the remaining cotyledon. Illumination of the cotyledonary lamina did not participate in the phototropic curvature of the hypocotyl, so long as the lamina was free to reorient to face the beam. When the lamina was continuously exposed to vectorial photoexcitation, elongation of the hypocotyl on the side carrying the cotyledon could be enhanced, or inhibited, depending on the direction of the beam striking its lamina.

Variation in Wolbachia effects on Aedes mosquitoes as a determinant of invasiveness and vectorial capacity.

PubMed

King, Jessica G; Souto-Maior, Caetano; Sartori, Larissa M; Maciel-de-Freitas, Rafael; Gomes, M Gabriela M

2018-04-16

Wolbachia has been introduced into Aedes aegypti mosquitoes to control the spread of arboviruses, such as dengue, chikungunya and Zika. Studies showed that certain Wolbachia strains (such as wMel) reduce replication of dengue viruses in the laboratory, prompting the release of mosquitoes carrying the bacterium into the field, where vectorial capacity can be realistically assessed in relation to native non-carriers. Here we apply a new analysis to two published datasets, and show that wMel increases the mean and the variance in Ae. aegypti susceptibility to dengue infection when introgressed into Brazil and Vietnam genetic backgrounds. In the absence of other processes, higher mean susceptibility should lead to enhanced viral transmission. The increase in variance, however, widens the basis for selection imposed by unexplored natural forces, retaining the potential for reducing transmission overall.

Nanoscale Membrane Curvature detected by Polarized Localization Microscopy

NASA Astrophysics Data System (ADS)

Kelly, Christopher; Maarouf, Abir; Woodward, Xinxin

Nanoscale membrane curvature is a necessary component of countless cellular processes. Here we present Polarized Localization Microscopy (PLM), a super-resolution optical imaging technique that enables the detection of nanoscale membrane curvature with order-of-magnitude improvements over comparable optical techniques. PLM combines the advantages of polarized total internal reflection fluorescence microscopy and fluorescence localization microscopy to reveal single-fluorophore locations and orientations without reducing localization precision by point spread function manipulation. PLM resolved nanoscale membrane curvature of a supported lipid bilayer draped over polystyrene nanoparticles on a glass coverslip, thus creating a model membrane with coexisting flat and curved regions and membrane radii of curvature as small as 20 nm. Further, PLM provides single-molecule trajectories and the aggregation of curvature-inducing proteins with super-resolution to reveal the correlated effects of membrane curvature, dynamics, and molecular sorting. For example, cholera toxin subunit B has been observed to induce nanoscale membrane budding and concentrate at the bud neck. PLM reveals a previously hidden and critical information of membrane topology.

Vectorial capacity and vector control: reconsidering sensitivity to parameters for malaria elimination.

PubMed

Brady, Oliver J; Godfray, H Charles J; Tatem, Andrew J; Gething, Peter W; Cohen, Justin M; McKenzie, F Ellis; Perkins, T Alex; Reiner, Robert C; Tusting, Lucy S; Sinka, Marianne E; Moyes, Catherine L; Eckhoff, Philip A; Scott, Thomas W; Lindsay, Steven W; Hay, Simon I; Smith, David L

2016-02-01

Major gains have been made in reducing malaria transmission in many parts of the world, principally by scaling-up coverage with long-lasting insecticidal nets and indoor residual spraying. Historically, choice of vector control intervention has been largely guided by a parameter sensitivity analysis of George Macdonald's theory of vectorial capacity that suggested prioritizing methods that kill adult mosquitoes. While this advice has been highly successful for transmission suppression, there is a need to revisit these arguments as policymakers in certain areas consider which combinations of interventions are required to eliminate malaria. Using analytical solutions to updated equations for vectorial capacity we build on previous work to show that, while adult killing methods can be highly effective under many circumstances, other vector control methods are frequently required to fill effective coverage gaps. These can arise due to pre-existing or developing mosquito physiological and behavioral refractoriness but also due to additive changes in the relative importance of different vector species for transmission. Furthermore, the optimal combination of interventions will depend on the operational constraints and costs associated with reaching high coverage levels with each intervention. Reaching specific policy goals, such as elimination, in defined contexts requires increasingly non-generic advice from modelling. Our results emphasize the importance of measuring baseline epidemiology, intervention coverage, vector ecology and program operational constraints in predicting expected outcomes with different combinations of interventions. © The Author 2016. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene.

Printing-assisted surface modifications of patterned ultrafiltration membranes

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

Wardrip, Nathaniel C.; Dsouza, Melissa; Urgun-Demirtas, Meltem

Understanding and restricting microbial surface attachment will enhance wastewater treatment with membranes. We report a maskless lithographic patterning technique for the generation of patterned polymer coatings on ultrafiltration membranes. Polyethylene glycol, zwitterionic, or negatively charged hydrophilic polymer compositions in parallel- or perpendicular-striped patterns with respect to feed flow were evaluated using wastewater. Membrane fouling was dependent on the orientation and chemical composition of the coatings. Modifications reduced alpha diversity in the attached microbial community (Shannon indices decreased from 2.63 to 1.89) which nevertheless increased with filtration time. Sphingomonas species, which condition membrane surfaces and facilitate cellular adhesion, were depleted inmore » all modified membranes. Microbial community structure was significantly different between control, different patterns, and different chemistries. Lastly, this study broadens the tools for surface modification of membranes with polymer coatings and for understanding and optimization of antifouling surfaces.« less

Probing Membrane Order and Topography in Supported Lipid Bilayers by Combined Polarized Total Internal Reflection Fluorescence-Atomic Force Microscopy

PubMed Central

Oreopoulos, John; Yip, Christopher M.

2009-01-01

Determining the local structure, dynamics, and conformational requirements for protein-protein and protein-lipid interactions in membranes is critical to understanding biological processes ranging from signaling to the translocating and membranolytic action of antimicrobial peptides. We report here the application of a combined polarized total internal reflection fluorescence microscopy-in situ atomic force microscopy platform. This platform's ability to image membrane orientational order was demonstrated on DOPC/DSPC/cholesterol model membranes containing the fluorescent membrane probe, DiI-C20 or BODIPY-PC. Spatially resolved order parameters and fluorophore tilt angles extracted from the polarized total internal reflection fluorescence microscopy images were in good agreement with the topographical details resolved by in situ atomic force microscopy, portending use of this technique for high-resolution characterization of membrane domain structures and peptide-membrane interactions. PMID:19254557

Printing-assisted surface modifications of patterned ultrafiltration membranes

DOE PAGES

Wardrip, Nathaniel C.; Dsouza, Melissa; Urgun-Demirtas, Meltem; ...

2016-10-17

Understanding and restricting microbial surface attachment will enhance wastewater treatment with membranes. We report a maskless lithographic patterning technique for the generation of patterned polymer coatings on ultrafiltration membranes. Polyethylene glycol, zwitterionic, or negatively charged hydrophilic polymer compositions in parallel- or perpendicular-striped patterns with respect to feed flow were evaluated using wastewater. Membrane fouling was dependent on the orientation and chemical composition of the coatings. Modifications reduced alpha diversity in the attached microbial community (Shannon indices decreased from 2.63 to 1.89) which nevertheless increased with filtration time. Sphingomonas species, which condition membrane surfaces and facilitate cellular adhesion, were depleted inmore » all modified membranes. Microbial community structure was significantly different between control, different patterns, and different chemistries. Lastly, this study broadens the tools for surface modification of membranes with polymer coatings and for understanding and optimization of antifouling surfaces.« less

[Differences in growth and ontogenetic development of plants grown in the Earth gravitational field in the natural and inverse orientation].

PubMed

Smolianina, S O; Berkovich, Iu A; Krivobok, N M; Ivanov, V B

2003-01-01

Wheat plants Triticum aestivum L., Apogee cultivar, were grown in the natural and inverse orientation of the Earth gravitational field. Special vegetation containers with double bottom were used for the cultivation. The upper bottom made of porous titanium served as a hydrophilic porous membrane stabilizing aquatic potential in the root-inhabited zone at a given level. Normal plants yielding viable seeds were obtained for both natural and inverse orientation. In our experiments, the inverse orientation induced dry weight accumulation by the plants as well as development of productive tillering shoots and increased the shoot-root dry weight ratio.

NSAID-derived gamma-secretase modulators. Part III: Membrane anchoring.

PubMed

Baumann, Stefanie; Höttecke, Nicole; Schubenel, Robert; Baumann, Karlheinz; Schmidt, Boris

2009-12-15

Selective lowering of Abeta(42) levels with small-molecule substrate targeting gamma-secretase modulators (sGSMs), such as some non-steroidal anti-inflammatory drugs, is a promising therapeutic approach for Alzheimer's disease. Here we present N-substituted carbazole- and O-substituted fenofibrate-derived sGSMs and their activity data. Seven out of 19 screened compounds exhibited promising activity against Abeta(42) secretion at a low micromolar level. We presume that the sGSMs interact with lys624 at the membrane interface and that the lipophilic substituents anchor the compound orientation in the membrane.

A study on the interactions of Aurein 2.5 with bacterial membranes.

PubMed

Dennison, Sarah R; Morton, Leslie H G; Shorrocks, Andrea J; Harris, Frederick; Phoenix, David A

2009-02-01

Aurein 2.5 (GLFDIVKKVVGAFGSL-NH(2)) is an uncharacterised antimicrobial peptide. At an air/water interface, it exhibited strong surface activity (maximal surface pressure 25mNm(-1)) and molecular areas consistent with the adoption of alpha-helical structure orientated either perpendicular (1.72nm(2)molecule(-1)) or parallel (3.6nm(2)molecule(-1)) to the interface. Aurein 2.5 was strongly antibacterial, exhibiting a minimum inhibitory concentration (MIC) of 30microM against Bacillus subtilis and Escherichia coli. The peptide induced maximal surface pressure changes of 9mNm(-1) and 5mNm(-1), respectively, in monolayers mimicking membranes of these organisms whilst compression isotherm analysis of these monolayers showed DeltaG(Mix)>0, indicating destabilisation by Aurein 2.5. These combined data suggested that toxicity of the peptide to these organisms may involve membrane invasion via the use of oblique orientated alpha-helical structure. The peptide induced strong, comparable maximal surface changes in monolayers of DOPG (7.5mNm(-1)) and DOPE monolayers (6mNm(-1)) suggesting that the membrane interactions of Aurein 2.5 were driven by amphiphilicity rather than electrostatic interaction. Based on these data, it was suggested that the differing ability of Aurein 2.5 to insert into membranes of B. subtilis and E. coli was probably related to membrane-based factors such as differences in lipid packing characteristics. The peptide was active against both sessile E. coli and Staphylococcus aureus with an MIC of 125microM. The broad-spectrum antibacterial activity and non-specific modes of membrane action used by Aurein 2.5 suggested use as an anti-biofilm agent such as in the decontamination of medical devices.

The Role of Protein-Protein and Protein-Membrane Interactions on P450 Function

PubMed Central

Scott, Emily E.; Wolf, C. Roland; Otyepka, Michal; Humphreys, Sara C.; Reed, James R.; Henderson, Colin J.; McLaughlin, Lesley A.; Paloncýová, Markéta; Navrátilová, Veronika; Berka, Karel; Anzenbacher, Pavel; Dahal, Upendra P.; Barnaba, Carlo; Brozik, James A.; Jones, Jeffrey P.; Estrada, D. Fernando; Laurence, Jennifer S.; Park, Ji Won

2016-01-01

This symposium summary, sponsored by the ASPET, was held at Experimental Biology 2015 on March 29, 2015, in Boston, Massachusetts. The symposium focused on: 1) the interactions of cytochrome P450s (P450s) with their redox partners; and 2) the role of the lipid membrane in their orientation and stabilization. Two presentations discussed the interactions of P450s with NADPH-P450 reductase (CPR) and cytochrome b5. First, solution nuclear magnetic resonance was used to compare the protein interactions that facilitated either the hydroxylase or lyase activities of CYP17A1. The lyase interaction was stimulated by the presence of b5 and 17α-hydroxypregnenolone, whereas the hydroxylase reaction was predominant in the absence of b5. The role of b5 was also shown in vivo by selective hepatic knockout of b5 from mice expressing CYP3A4 and CYP2D6; the lack of b5 caused a decrease in the clearance of several substrates. The role of the membrane on P450 orientation was examined using computational methods, showing that the proximal region of the P450 molecule faced the aqueous phase. The distal region, containing the substrate-access channel, was associated with the membrane. The interaction of NADPH-P450 reductase (CPR) with the membrane was also described, showing the ability of CPR to “helicopter” above the membrane. Finally, the endoplasmic reticulum (ER) was shown to be heterogeneous, having ordered membrane regions containing cholesterol and more disordered regions. Interestingly, two closely related P450s, CYP1A1 and CYP1A2, resided in different regions of the ER. The structural characteristics of their localization were examined. These studies emphasize the importance of P450 protein organization to their function. PMID:26851242

Optimization of differentiation time of mesenchymal-stem-cell to tenocyte under a cyclic stretching with a microgrooved culture membrane and selected measurement cells.

PubMed

Morita, Yasuyuki; Yamashita, Takahiro; Toku, Toku; Ju, Yang

2018-01-01

There is a need for efficient stem cell-to-tenocyte differentiation techniques for tendon tissue engineering. More than 1 week is required for tenogenic differentiation with chemical stimuli, including co-culturing. Research has begun to examine the utility of mechanical stimuli, which reduces the differentiation time to several days. However, the precise length of time required to differentiate human bone marrow-derived mesenchymal stem cells (hBMSCs) into tenocytes has not been clarified. Understanding the precise time required is important for future tissue engineering projects. Therefore, in this study, a method was developed to more precisely determine the length of time required to differentiate hBMSCs into tenocytes with cyclic stretching stimulus. First, it had to be determined how stretching stimulation affected the cells. Microgrooved culture membranes were used to suppress cell orientation behavior. Then, only cells oriented parallel to the microgrooves were selected and evaluated for protein synthesis levels for differentiation. The results revealed that growing cells on the microgrooved membrane and selecting optimally-oriented cells for measurement improved the accuracy of the differentiation evaluation, and that hBMSCs differentiated into tenocytes in approximately 10 h. The differentiation time corresponded to the time required for cellular cytoskeleton reorganization and cellular morphology alterations. This suggests that cells, when subjected to mechanical stimulus, secrete mRNAs and proteins for both cytoskeleton reorganization and differentiation.

Fatty acyl chain order in lecithin model membranes determined from proton magnetic resonance.

PubMed

Bloom, M; Burnell, E E; MacKay, A L; Nichol, C P; Valic, M I; Weeks, G

1978-12-26

Proton magnetic resonance (1H NMR) has been used to compare the local orientational order of acyl chains in phospholipid bilayers of multilamellar and small sonicated vesicular membranes of dipalmitoyllecithin (DPL) at 50 degrees C and egg yolk lecithin (EYL) at 31 degrees C. The orientational order of the multilamellar systems was characterized using deuterium magnetic resonance order parameters and 1H NMR second moments. 1H NMR line shapes in the vesicle samples were calculated using vesicle size distributions, determined directly using electron microscopy, and a theory of motional narrowing, which takes into account the symmetry properties of the bilayer systems. The predicted non-Lorentzian line shapes and widths were found to be in good agreement with experimental results, indicating that the local orientational order (called "packing" by many workers) in the bilayers of small vesicles and in multilamellar membranes is substantially the same. This results was found to be true not only for the largest 1H NMR line associated with the nonterminal methylene protons but also for the resolved 1H NMR lines due to the alpha-CH2 and the terminal CH3 positions on the acyl chain. Analysis of the vesicle 1H NMR spectra of EYL taken with different medium viscosities yielded a value of approximately 4 X 10(-8) cm2 s-1 for the lateral diffusion constant of the phospholipid molecules at 31 degrees C.

Ion/proton-conducting apparatus and method

DOEpatents

Yates, Matthew; Xue, Wei

2014-12-23

A c-axis-oriented HAP thin film synthesized by seeded growth on a palladium hydrogen membrane substrate. An exemplary synthetic process includes electrochemical seeding on the substrate, and secondary and tertiary hydrothermal treatments under conditions that favor growth along c-axes and a-axes in sequence. By adjusting corresponding synthetic conditions, an HAP this film can be grown to a controllable thickness with a dense coverage on the underlying substrate. The thin films have relatively high proton conductivity under hydrogen atmosphere and high temperature conditions. The c-axis oriented films may be integrated into fuel cells for application in the intermediate temperature range of 200-600.degree. C. The electrochemical-hydrothermal deposition technique may be applied to create other oriented crystal materials having optimized properties, useful for separations and catalysis as well as electronic and electrochemical applications, electrochemical membrane reactors, and in chemical sensors. Additional high-density and gas-tight HAP film compositions may be deposited using a two-step deposition method that includes an electrochemical deposition method followed by a hydrothermal deposition method. The two-step method uses a single hydrothermal deposition solution composition. The method may be used to deposit HAP films including but not limited to at least doped HAP films, and more particularly including carbonated HAP films. In addition, the high-density and gas-tight HAP films may be used in proton exchange membrane fuel cells.

Difficulties that Students Face with Two-Dimensional Motion

ERIC Educational Resources Information Center

Mihas, P.; Gemousakakis, T.

2007-01-01

Some difficulties that students face with two-dimensional motion are addressed. The difficulties addressed are the vectorial representation of velocity, acceleration and force, the force-energy theorem and the understanding of the radius of curvature.

Mechanical Stimulation of Stem Cells Using Cyclic Uniaxial Strain

PubMed Central

Kurpinski, Kyle; Li, Song

2007-01-01

The role of mechanical forces in the development and maintenance of biological tissues is well documented, including several mechanically regulated phenomena such as bone remodeling, muscular hypertrophy, and smooth muscle cell plasticity. However, the forces involved are often extremely complex and difficult to monitor and control in vivo. To better investigate the effects of mechanical forces on cells, we have developed an in vitro method for applying uniaxial cyclic tensile strain to adherent cells cultured on elastic membranes. This method utilizes a custom-designed bioreactor with a motorized cam-rotor system to apply the desired force. Here we present a step-by-step video protocol demonstrating how to assemble the various components of each "stretch chamber", including, in this case, a silicone membrane with micropatterned topography to orient the cells with the direction of the strain. We also describe procedures for sterilizing the chambers, seeding cells onto the membrane, latching the chamber into the bioreactor, and adjusting the mechanical parameters (i.e. magnitude and rate of strain). The procedures outlined in this particular protocol are specific for seeding human mesenchymal stem cells onto silicone membranes with 10 µm wide channels oriented parallel to the direction of strain. However, the methods and materials presented in this system are flexible enough to accommodate a number of variations on this theme: strain rate, magnitude, duration, cell type, membrane topography, membrane coating, etc. can all be tailored to the desired application or outcome. This is a robust method for investigating the effects of uniaxial tensile strain applied to cells in vitro. PMID:18997890

Comparative Study of MIL-96(Al) as Continuous Metal-Organic Frameworks Layer and Mixed-Matrix Membrane.

PubMed

Knebel, Alexander; Friebe, Sebastian; Bigall, Nadja Carola; Benzaqui, Marvin; Serre, Christian; Caro, Jürgen

2016-03-23

MIL-96(Al) layers were prepared as supported metal-organic frameworks membrane via reactive seeding using the α-alumina support as the Al source for the formation of the MIL-96(Al) seeds. Depending on the solvent mixture employed during seed formation, two different crystal morphologies, with different orientation of the transport-active channels, have been formed. This crystal orientation and habit is predefined by the seed crystals and is kept in the subsequent growth of the seeds to continuous layers. In the gas separation of an equimolar H2/CO2 mixture, the hydrogen permeability of the two supported MIL-96(Al) layers was found to be highly dependent on the crystal morphology and the accompanied channel orientation in the layer. In addition to the neat supported MIL-96(Al) membrane layers, mixed-matrix membranes (MMMs, 10 wt % filler loading) as a composite of MIL-96(Al) particles as filler in a continuous Matrimid polymer phase have been prepared. Five particle sizes of MIL-96(Al) between 3.2 μm and 55 nm were synthesized. In the preparation of the MIL-96(Al)/Matrimid MMM (10 wt % filler loading), the following preparation problems have been identified: The bigger micrometer-sized MIL-96(Al) crystals show a trend toward sedimentation during casting of the MMM, whereas for nanoparticles aggregation and recrystallization to micrometer-sized MIL-96(Al) crystals has been observed. Because of these preparation problems for MMM, the neat supported MIL-96(Al) layers show a relatively high H2/CO2 selectivity (≈9) and a hydrogen permeance approximately 2 magnitudes higher than that of the best MMM.

Transport rates of a glutamate transporter homologue are influenced by the lipid bilayer.

PubMed

McIlwain, Benjamin C; Vandenberg, Robert J; Ryan, Renae M

2015-04-10

The aspartate transporter from Pyrococcus horikoshii (GltPh) is a model for the structure of the SLC1 family of amino acid transporters. Crystal structures of GltPh provide insight into mechanisms of ion coupling and substrate transport; however, structures have been solved in the absence of a lipid bilayer so they provide limited information regarding interactions that occur between the protein and lipids of the membrane. Here, we investigated the effect of the lipid environment on aspartate transport by reconstituting GltPh into liposomes of defined lipid composition where the primary lipid is phosphatidylethanolamine (PE) or its methyl derivatives. We showed that the rate of aspartate transport and the transmembrane orientation of GltPh were influenced by the primary lipid in the liposomes. In PE liposomes, we observed the highest transport rate and showed that 85% of the transporters were orientated right-side out, whereas in trimethyl PE liposomes, 50% of transporters were right-side out, and we observed a 4-fold reduction in transport rate. Differences in orientation can only partially explain the lipid composition effect on transport rate. Crystal structures of GltPh revealed a tyrosine residue (Tyr-33) that we propose interacts with lipid headgroups during the transport cycle. Based on site-directed mutagenesis, we propose that a cation-π interaction between Tyr-33 and the lipid headgroups can influence conformational flexibility of the trimerization domain and thus the rate of transport. These results provide a specific example of how interactions between membrane lipids and membrane-bound proteins can influence function and highlight the importance of the role of the membrane in transporter function. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

A Study of Parameters Affecting Fibroblast Morphology in Response to an Applied Mechanical Force

NASA Technical Reports Server (NTRS)

Grymes, Rosalind A.; Sawyer, Christine

1994-01-01

A precisely controlled stretch/relaxation regimen (20% elongation at 6.6 cycles/min) was applied to normal human fetal, neonatal and aged dermal fibroblasts cultured on flexible membranes. Culture conditions included poly (NH2) or collagen type I coated substrate membranes; control cultures were grown on the same pliable material in the absence of applied stretch. Direct observation and immunofluorescence analyses revealed a progressive change in cell body orientation limited to the stretched dermal fibroblast cultures. Monolayers gradually (over 4 days) acquired a symmetric, radial distribution equivalent to the biaxial array of the applied force. At high seeding density, alignment was inhibited in the fetal cell cultures. This cell strain required collagen type I coating for optimal attachment to the flexible membrane, preferring growth in three-dimensional cell 'balls' on the poly(NH2) coated substrate. Neonatal cells also required the collagen type I coating, but both neonatal and aged dermal fibroblasts aligned efficiently at all seeding densities examined. The randomly oriented neonatal cells on the unstretched control membranes spontaneously detached at confluence, as a single cell sheet. Their aligned counterparts did not detach until the applied stretch stimulus was removed. Low concentrations of cytochalasin D (62.5 ng/ml) disrupted the stretch-related alignment response. Rhodamine phalloidin staining visualized fewer actin stress fibers in stretched, aligned cells than in controls. Both intercellular interactions and cytoskeletal integrity mediate the response to mechanical strain. Normal rabbit corneal stroma fibroblasts (NRC) were also analyzed, and failed to orient under these conditions. This cell type may require a different regimen, or a longer time period, to demonstrate alignment behavior. Supported by NASA Space Biology RTOP 199-40-22 and the NASA-ARC Director's Discretionary Fund.

Transport Rates of a Glutamate Transporter Homologue Are Influenced by the Lipid Bilayer*

PubMed Central

McIlwain, Benjamin C.; Vandenberg, Robert J.; Ryan, Renae M.

2015-01-01

The aspartate transporter from Pyrococcus horikoshii (GltPh) is a model for the structure of the SLC1 family of amino acid transporters. Crystal structures of GltPh provide insight into mechanisms of ion coupling and substrate transport; however, structures have been solved in the absence of a lipid bilayer so they provide limited information regarding interactions that occur between the protein and lipids of the membrane. Here, we investigated the effect of the lipid environment on aspartate transport by reconstituting GltPh into liposomes of defined lipid composition where the primary lipid is phosphatidylethanolamine (PE) or its methyl derivatives. We showed that the rate of aspartate transport and the transmembrane orientation of GltPh were influenced by the primary lipid in the liposomes. In PE liposomes, we observed the highest transport rate and showed that 85% of the transporters were orientated right-side out, whereas in trimethyl PE liposomes, 50% of transporters were right-side out, and we observed a 4-fold reduction in transport rate. Differences in orientation can only partially explain the lipid composition effect on transport rate. Crystal structures of GltPh revealed a tyrosine residue (Tyr-33) that we propose interacts with lipid headgroups during the transport cycle. Based on site-directed mutagenesis, we propose that a cation-π interaction between Tyr-33 and the lipid headgroups can influence conformational flexibility of the trimerization domain and thus the rate of transport. These results provide a specific example of how interactions between membrane lipids and membrane-bound proteins can influence function and highlight the importance of the role of the membrane in transporter function. PMID:25713135

Structural features and lipid binding domain of tubulin on biomimetic mitochondrial membranes

PubMed Central

Hoogerheide, David P.; Noskov, Sergei Y.; Jacobs, Daniel; Bergdoll, Lucie; Silin, Vitalii; Worcester, David L.; Abramson, Jeff; Nanda, Hirsh; Rostovtseva, Tatiana K.; Bezrukov, Sergey M.

2017-01-01

Dimeric tubulin, an abundant water-soluble cytosolic protein known primarily for its role in the cytoskeleton, is routinely found to be associated with mitochondrial outer membranes, although the structure and physiological role of mitochondria-bound tubulin are still unknown. There is also no consensus on whether tubulin is a peripheral membrane protein or is integrated into the outer mitochondrial membrane. Here the results of five independent techniques—surface plasmon resonance, electrochemical impedance spectroscopy, bilayer overtone analysis, neutron reflectometry, and molecular dynamics simulations—suggest that α-tubulin’s amphipathic helix H10 is responsible for peripheral binding of dimeric tubulin to biomimetic “mitochondrial” membranes in a manner that differentiates between the two primary lipid headgroups found in mitochondrial membranes, phosphatidylethanolamine and phosphatidylcholine. The identification of the tubulin dimer orientation and membrane-binding domain represents an essential step toward our understanding of the complex mechanisms by which tubulin interacts with integral proteins of the mitochondrial outer membrane and is important for the structure-inspired design of tubulin-targeting agents. PMID:28420794

Protein targeting and integration signal for the chloroplastic outer envelope membrane.

PubMed Central

Li, H M; Chen, L J

1996-01-01

Most proteins in chloroplasts are encoded by the nuclear genome and synthesized in the cytosol. With the exception of most quter envelope membrane proteins, nuclear-encoded chloroplastic proteins are synthesized with N-terminal extensions that contain the chloroplast targeting information of these proteins. Most outer membrane proteins, however, are synthesized without extensions in the cytosol. Therefore, it is not clear where the chloroplastic outer membrane targeting information resides within these polypeptides. We have analyzed a chloroplastic outer membrane protein, OEP14 (outer envelope membrane protein of 14 kD, previously named OM14), and localized its outer membrane targeting and integration signal to the first 30 amino acids of the protein. This signal consists of a positively charged N-terminal portion followed by a hydrophobic core, bearing resemblance to the signal peptides of proteins targeted to the endoplasmic reticulum. However, a chimeric protein containing this signal fused to a passenger protein did not integrate into the endoplasmic reticulum membrane. Furthermore, membrane topology analysis indicated that the signal inserts into the chloroplastic outer membrane in an orientation opposite to that predicted by the "positive inside" rule. PMID:8953775

Conductivity of an inverse lyotropic lamellar phase under shear flow

NASA Astrophysics Data System (ADS)

Panizza, P.; Soubiran, L.; Coulon, C.; Roux, D.

2001-08-01

We report conductivity measurements on solutions of closed compact monodisperse multilamellar vesicles (the so-called ``onion texture'') formed by shearing an inverse lyotropic lamellar Lα phase. The conductivity measured in different directions as a function of the applied shear rate reveals a small anisotropy of the onion structure due to the existence of free oriented membranes. The results are analyzed in terms of a simple model that allows one to deduce the conductivity tensor of the Lα phase itself and the proportion of free oriented membranes. The variation of these two parameters is measured along a dilution line and discussed. The high value of the conductivity perpendicular to the layers with respect to that of solvent suggests the existence of a mechanism of ionic transport through the insulating solvent.

Metal-Organic Framework (MOF) Nanorods, Nanotubes, and Nanowires.

PubMed

Arbulu, Roberto C; Jiang, Ying-Bing; Peterson, Eric J; Qin, Yang

2018-05-14

New mechanisms for the controlled growth of one-dimensional (1D) metal-organic framework (MOF) nano- and superstructures under size-confinement and surface-directing effects have been discovered. Through applying interfacial synthesis templated by track-etched polycarbonate (PCTE) membranes, congruent polycrystalline zeolitic imidazolate framework-8 (ZIF-8) solid nanorods and hollow nanotubes were found to form within 100 nm membrane pores, while single crystalline ZIF-8 nanowires grew inside 30 nm pores, all of which possess large aspect ratios up to 60 and show preferential crystal orientation with the {100} planes aligned parallel to the long axis of the pore. Our findings provide a generalizable method for controlling size, morphology, and lattice orientation of MOF nanomaterials. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Mosquito biology. Evolution of sexual traits influencing vectorial capacity in anopheline mosquitoes.

PubMed

Mitchell, Sara N; Kakani, Evdoxia G; South, Adam; Howell, Paul I; Waterhouse, Robert M; Catteruccia, Flaminia

2015-02-27

The availability of genome sequences from 16 anopheline species provides unprecedented opportunities to study the evolution of reproductive traits relevant for malaria transmission. In Anopheles gambiae, a likely candidate for sexual selection is male 20-hydroxyecdysone (20E). Sexual transfer of this steroid hormone as part of a mating plug dramatically changes female physiological processes intimately tied to vectorial capacity. By combining phenotypic studies with ancestral state reconstructions and phylogenetic analyses, we show that mating plug transfer and male 20E synthesis are both derived characters that have coevolved in anophelines, driving the adaptation of a female 20E-interacting protein that promotes oogenesis via mechanisms also favoring Plasmodium survival. Our data reveal coevolutionary dynamics of reproductive traits between the sexes likely to have shaped the ability of anophelines to transmit malaria. Copyright © 2015, American Association for the Advancement of Science.

Thickness and angular dependent magnetic anisotropy of La0.67Sr0.33MnO3 thin films by Vectorial Magneto Optical Kerr Magnetometry

NASA Astrophysics Data System (ADS)

Chaluvadi, S. K.; Perna, P.; Ajejas, F.; Camarero, J.; Pautrat, A.; Flament, S.; Méchin, L.

2017-10-01

We investigate the in-plane magnetic anisotropy in La0.67Sr0.33MnO3 thin films grown on SrTiO3 (001) substrate using angular dependent room temperature Vectorial Magneto-Optical Kerr Magnetometry. The experimental data reveals that the magnetic anisotropy symmetry landscape significantly changes depending upon the strain and thickness. At low film thickness (12 and 25 nm) the dominant uniaxial anisotropy is due to interface effects, step edges due to mis-cut angle of SrTiO3 substrate. At intermediate thickness, the magnetic anisotropy presents a competition between magnetocrystalline (biaxial) and substrate step induced (uniaxial) anisotropy. Depending upon their relative strengths, a profound biaxial or uniaxial or mixed anisotropy is favoured. Above the critical thickness, magnetocrystalline anisotropy dominates all other effects and shows a biaxial anisotropy.

Bidimensional nano-optomechanics and topological backaction in a non-conservative radiation force field.

PubMed

Gloppe, A; Verlot, P; Dupont-Ferrier, E; Siria, A; Poncharal, P; Bachelier, G; Vincent, P; Arcizet, O

2014-11-01

Optomechanics, which explores the fundamental coupling between light and mechanical motion, has made important advances in manipulating macroscopic mechanical oscillators down to the quantum level. However, dynamical effects related to the vectorial nature of the optomechanical interaction remain to be investigated. Here we study a nanowire with subwavelength dimensions coupled strongly to a tightly focused beam of light, enabling an ultrasensitive readout of the nanoresonator dynamics. We determine experimentally the vectorial structure of the optomechanical interaction and demonstrate that a bidimensional dynamical backaction governs the nanowire dynamics. Moreover, the spatial topology of the optomechanical interaction is responsible for novel canonical signatures of strong coupling between mechanical modes, which leads to a topological instability that underlies the non-conservative nature of the optomechanical interaction. These results have a universal character and illustrate the increased sensitivity of nanomechanical devices towards spatially varying interactions, opening fundamental perspectives in nanomechanics, optomechanics, ultrasensitive scanning force microscopy and nano-optics.

Manipulation of dielectric Rayleigh particles using highly focused elliptically polarized vector fields.

PubMed

Gu, Bing; Xu, Danfeng; Rui, Guanghao; Lian, Meng; Cui, Yiping; Zhan, Qiwen

2015-09-20

Generation of vectorial optical fields with arbitrary polarization distribution is of great interest in areas where exotic optical fields are desired. In this work, we experimentally demonstrate the versatile generation of linearly polarized vector fields, elliptically polarized vector fields, and circularly polarized vortex beams through introducing attenuators in a common-path interferometer. By means of Richards-Wolf vectorial diffraction method, the characteristics of the highly focused elliptically polarized vector fields are studied. The optical force and torque on a dielectric Rayleigh particle produced by these tightly focused vector fields are calculated and exploited for the stable trapping of dielectric Rayleigh particles. It is shown that the additional degree of freedom provided by the elliptically polarized vector field allows one to control the spatial structure of polarization, to engineer the focusing field, and to tailor the optical force and torque on a dielectric Rayleigh particle.

Orphan Kinesin NOD Lacks Motile Properties But Does Possess a Microtubule-stimulated ATPase Activity

PubMed Central

Matthies, Heinrich J.G.; Baskin, Ronald J.; Hawley, R. Scott

2001-01-01

NOD is a Drosophila chromosome-associated kinesin-like protein that does not fall into the chromokinesin subfamily. Although NOD lacks residues known to be critical for kinesin function, we show that microtubules activate the ATPase activity of NOD >2000-fold. Biochemical and genetic analysis of two genetically identified mutations of NOD (NODDTW and NOD“DR2”) demonstrates that this allosteric activation is critical for the function of NOD in vivo. However, several lines of evidence indicate that this ATPase activity is not coupled to vectorial transport, including 1) NOD does not produce microtubule gliding; and 2) the substitution of a single amino acid in the Drosophila kinesin heavy chain with the analogous amino acid in NOD results in a drastic inhibition of motility. We suggest that the microtubule-activated ATPase activity of NOD provides transient attachments of chromosomes to microtubules rather than producing vectorial transport. PMID:11739796

[Assessment of malaria resurgence risk in Morocco. Study of the vectorial capacity of Anopheles labranchiae in a rice cultivation area in the north of the country].

PubMed

Faraj, C; Ouahabi, S; Adlaoui, E; Boccolini, D; Romi, R; El Aouad, R

2008-12-01

To assess the malaria reintroduction risk in Morocco, we analyzed the malariogenic potential of a rice cultivation area in the north of the country. Our results showed that the receptivity of this area is very high during all the period of the rice cultivation, from May to October, the vectorial capacity of An. labranchiae, malaria vector in Morocco, is considerably high during the summer which corresponds to the rice cultivation period. The risk of autochthonous malaria resumption is important because of the possible presence of gametocytes carriers in the last malaria focus which is bordering the study area. The risk of a tropical malaria introduction is unimportant seen the low vulnerability of the area and the uncertain competence of its vectors considered. However, this risk must be considered with a more attention.

General imaging of advanced 3D mask objects based on the fully-vectorial extended Nijboer-Zernike (ENZ) theory

NASA Astrophysics Data System (ADS)

van Haver, Sven; Janssen, Olaf T. A.; Braat, Joseph J. M.; Janssen, Augustus J. E. M.; Urbach, H. Paul; Pereira, Silvania F.

2008-03-01

In this paper we introduce a new mask imaging algorithm that is based on the source point integration method (or Abbe method). The method presented here distinguishes itself from existing methods by exploiting the through-focus imaging feature of the Extended Nijboer-Zernike (ENZ) theory of diffraction. An introduction to ENZ-theory and its application in general imaging is provided after which we describe the mask imaging scheme that can be derived from it. The remainder of the paper is devoted to illustrating the advantages of the new method over existing methods (Hopkins-based). To this extent several simulation results are included that illustrate advantages arising from: the accurate incorporation of isolated structures, the rigorous treatment of the object (mask topography) and the fully vectorial through-focus image formation of the ENZ-based algorithm.

Cation-containing lipid membranes – experiment and md simulations

DOE PAGES

Kučerka, Norbert; Dushanov, Ermuhammas; Kholmurodov, Kholmirzo T.; ...

2017-11-27

Here, using small angle neutron diffraction and molecular dynamics simulations we studied the interactions between calcium (Ca 2+) or zinc (Zn 2+) cations, and oriented gel phase dipalmitoyl-phosphatidylcholine (DPPC) bilayers. For both cations studied at ~1:7 divalent metal ion to lipid molar ratio (Me2+:DPPC), bilayer thickness increased. Simulation results helped reveal subtle differences in the effects of the two cations on gel phase membranes.

Resolving the 3D spatial orientation of helix I in the closed state of the colicin E1 channel domain by FRET. Insights into the integration mechanism.

PubMed

Lugo, Miguel R; Ho, Derek; Merrill, A Rod

2016-10-15

Current evidence suggests that the closed-state membrane model for the channel-forming domain of colicin E1 involves eight amphipathic α-helices (helices I-VII and X) that adopt a two-dimensional arrangement on the membrane surface. Two central hydrophobic α-helices in colicin E1 (VIII and IX) adopt a transmembrane location-the umbrella model. Helices I and II have been shown to participate in the channel by forming a transmembrane segment (TM1) in the voltage-induced open channel state. Consequently, it is paramount to determine the relative location and orientation of helix I in the two-dimensional arrangement of the membrane. A new, low-resolution, three-dimensional model of the closed state of the colicin E1 channel was constructed based on FRET measurements between three naturally occurring Trp residues and three sites in helix I, in addition to previously reported FRET distances for the channel domain. Furthermore, a new mechanism for the channel integration process involving the transition of the soluble to membrane-bound form is presented based on a plethora of kinetic data for this process. Crown Copyright © 2016. Published by Elsevier Inc. All rights reserved.

Ultrasound assisted forward osmosis concentration of fruit juice and natural colorant.

PubMed

Chanukya, B S; Rastogi, Navin K

2017-01-01

The present study deals with the effect of higher and lower molecular weight compounds present in the feed on concentration polarization during forward osmosis concentration and its mitigation by the application of ultrasound. The effects of ultrasound on transmembrane water flux at different forward osmosis membrane orientations and different model feed solutions consisting of sucrose and pectin have also been evaluated. The feed containing sucrose and pectin subjected towards active layer of the membrane was found to be the most suitable orientation. The application of ultrasound (30kHz) significantly reduced the concentration polarization when the feed contains sucrose concentration up to 5%. Whereas, in case of feed containing 0.5% pectin, the ultrasound was not found to be effective in dislodging the gel layer formation resulting in severe external concentration polarization on the membrane surface. In comparison to the ordinary forward osmosis process, the ultrasound-assisted forward osmosis process resulted in higher water fluxes in case of sweet lime juice as well as rose extract containing anthocyanin. The degradation of rose anthocyanin due to ultrasound was found to be 1.82%. Application of ultrasound was found to be an effective way in mitigating concentration polarization on the forward osmosis membrane resulting in increased flux. Copyright © 2016 Elsevier B.V. All rights reserved.

Structure and function of the tetraheme cytochrome associated to the reaction center of Roseobacter denitrificans.

PubMed

Garcia, D; Richaud, P; Breton, J; Verméglio, A

1994-01-01

We have characterized the tetrahemic RC bound cytochrome isolated from the quasi-photosynthetic bacterium Roseobacter denitrificans in terms of absorption spectrum, redox property and orientation with respect to the membrane plane. The heme, designated H1, which possesses the highest redox midpoint potential (+290 mV), absorbs at 555 nm. Its plane makes an angle of 40 degrees with the membrane plane. The second high potential heme, H2 (+240 mV), peaks at 554 nm and makes a tilt of 55 degrees with the membrane. The two low potential hemes, L1 and L2, present a similar and rather high redox midpoint potential (+90 mV). They absorb at 553 nm and 550 nm. One of these hemes is oriented at 40 degrees while the other makes an angle of 90 degrees with the membrane plane. The soluble cytochrome c551 completes the cyclic electron transfer between the RC and the bc1 complex. Both the oxidation and the re-reduction of cytochrome c551 are diffusible processes. Under semi-aerobic conditions, one of the low potential hemes is photo-oxidized under illumination but only extremely slowly re-reduced. This explains the requirement of high aerobic conditions for growth of Roseobacter denitrificans cells in the light.

PEM/SPE fuel cell

DOEpatents

Grot, Stephen Andreas

1998-01-01

A PEM/SPE fuel cell including a membrane-electrode assembly (MEA) having a plurality of oriented filament embedded the face thereof for supporting the MEA and conducting current therefrom to contiguous electrode plates.

GLRS-R 2-colour retroreflector target design and predicted performance

NASA Technical Reports Server (NTRS)

Lund, Glenn

1993-01-01

This paper reports on the retroreflector ground-target design for the GLRS-R spaceborne dual-wavelength laser ranging system. The described passive design flows down from the requirements of high station autonomy, high global FOV (up to 60 degrees zenith angle), little or no multiple pulse returns, and adequate optical cross section for most ranging geometries. The proposed solution makes use of 5 hollow cube-corner retroreflectors of which one points to the zenith and the remaining four are inclined from the vertical at uniform azimuthal spacings. The need for fairly large (is approximately 10 cm) retroreflectors is expected (within turbulence limitations) to generate quite narrow diffraction lobes, thus placing non-trivial requirements on the vectorial accuracy of velocity aberration corrections. A good compromise solution is found by appropriately spoiling just one of the retroreflector dihedral angles from 90 degrees, thus generating two symmetrically oriented diffraction lobes in the return beam. The required spoil angles are found to have little dependence on ground target latitude. Various link budget analyses are presented, showing the influence of such factors as point-ahead optimization, turbulence, ranging angle, atmospheric visibility and ground target thermal deformations.

GLRS-R 2-colour retroreflector target design and predicted performance

NASA Astrophysics Data System (ADS)

Lund, Glenn

1993-06-01

This paper reports on the retroreflector ground-target design for the GLRS-R spaceborne dual-wavelength laser ranging system. The described passive design flows down from the requirements of high station autonomy, high global FOV (up to 60 degrees zenith angle), little or no multiple pulse returns, and adequate optical cross section for most ranging geometries. The proposed solution makes use of 5 hollow cube-corner retroreflectors of which one points to the zenith and the remaining four are inclined from the vertical at uniform azimuthal spacings. The need for fairly large (is approximately 10 cm) retroreflectors is expected (within turbulence limitations) to generate quite narrow diffraction lobes, thus placing non-trivial requirements on the vectorial accuracy of velocity aberration corrections. A good compromise solution is found by appropriately spoiling just one of the retroreflector dihedral angles from 90 degrees, thus generating two symmetrically oriented diffraction lobes in the return beam. The required spoil angles are found to have little dependence on ground target latitude. Various link budget analyses are presented, showing the influence of such factors as point-ahead optimization, turbulence, ranging angle, atmospheric visibility and ground target thermal deformations.

Early stages of clathrin aggregation at a membrane in coarse-grained simulations

NASA Astrophysics Data System (ADS)

Giani, M.; den Otter, W. K.; Briels, W. J.

2017-04-01

The self-assembly process of clathrin coated pits during endocytosis has been simulated by combining and extending coarse grained models of the clathrin triskelion, the adaptor protein AP2, and a flexible network membrane. The AP2's core, upon binding to membrane and cargo, releases a motif that can bind clathrin. In conditions where the core-membrane-cargo binding is weak, the binding of this motif to clathrin can result in a stable complex. We characterize the conditions and mechanisms resulting in the formation of clathrin lattices that curve the membrane, i.e., clathrin coated pits. The mechanical properties of the AP2 β linker appear crucial to the orientation of the curved clathrin lattice relative to the membrane, with wild-type short linkers giving rise to the inward curving buds enabling endocytosis while long linkers produce upside-down cages and outward curving bulges.

Crystal nuclei templated nanostructured membranes prepared by solvent crystallization and polymer migration

NASA Astrophysics Data System (ADS)

Wang, Bo; Ji, Jing; Li, Kang

2016-09-01

Currently, production of porous polymeric membranes for filtration is predominated by the phase-separation process. However, this method has reached its technological limit, and there have been no significant breakthrough over the last decade. Here we show, using polyvinylidene fluoride as a sample polymer, a new concept of membrane manufacturing by combining oriented green solvent crystallization and polymer migration is able to obtain high performance membranes with pure water permeation flux substantially higher than those with similar pore size prepared by conventional phase-separation processes. The new manufacturing procedure is governed by fewer operating parameters and is, thus, easier to control with reproducible results. Apart from the high water permeation flux, the prepared membranes also show excellent stable flux after fouling and superior mechanical properties of high pressure load and better abrasion resistance. These findings demonstrate the promise of a new concept for green manufacturing nanostructured polymeric membranes with high performances.

Highly oriented photosynthetic reaction centers generate a proton gradient in synthetic protocells

PubMed Central

Altamura, Emiliano; Milano, Francesco; Tangorra, Roberto R.; Trotta, Massimo; Omar, Omar Hassan; Stano, Pasquale

2017-01-01

Photosynthesis is responsible for the photochemical conversion of light into the chemical energy that fuels the planet Earth. The photochemical core of this process in all photosynthetic organisms is a transmembrane protein called the reaction center. In purple photosynthetic bacteria a simple version of this photoenzyme catalyzes the reduction of a quinone molecule, accompanied by the uptake of two protons from the cytoplasm. This results in the establishment of a proton concentration gradient across the lipid membrane, which can be ultimately harnessed to synthesize ATP. Herein we show that synthetic protocells, based on giant lipid vesicles embedding an oriented population of reaction centers, are capable of generating a photoinduced proton gradient across the membrane. Under continuous illumination, the protocells generate a gradient of 0.061 pH units per min, equivalent to a proton motive force of 3.6 mV⋅min−1. Remarkably, the facile reconstitution of the photosynthetic reaction center in the artificial lipid membrane, obtained by the droplet transfer method, paves the way for the construction of novel and more functional protocells for synthetic biology. PMID:28320948

A Rationally Designed, General Strategy for Membrane Orientation of Photoinduced Electron Transfer-Based Voltage-Sensitive Dyes.

PubMed

Kulkarni, Rishikesh U; Yin, Hang; Pourmandi, Narges; James, Feroz; Adil, Maroof M; Schaffer, David V; Wang, Yi; Miller, Evan W

2017-02-17

Voltage imaging with fluorescent dyes offers promise for interrogating the complex roles of membrane potential in coordinating the activity of neurons in the brain. Yet, low sensitivity often limits the broad applicability of optical voltage indicators. In this paper, we use molecular dynamics (MD) simulations to guide the design of new, ultrasensitive fluorescent voltage indicators that use photoinduced electron transfer (PeT) as a voltage-sensing switch. MD simulations predict an approximately 16% increase in voltage sensitivity resulting purely from improved alignment of dye with the membrane. We confirm this theoretical finding by synthesizing 9 new voltage-sensitive (VoltageFluor, or VF) dyes and establishing that all of them display the expected improvement of approximately 19%. This synergistic outworking of theory and experiment enabled computational and theoretical estimation of VF dye orientation in lipid bilayers and has yielded the most sensitive PeT-based VF dye to date. We use this new voltage indicator to monitor voltage spikes in neurons from rat hippocampus and human pluripotent-stem-cell-derived dopaminergic neurons.

Vectorial transport of bile salts across MDCK cells expressing both rat Na+-taurocholate cotransporting polypeptide and rat bile salt export pump.

PubMed

Mita, Sachiko; Suzuki, Hiroshi; Akita, Hidetaka; Stieger, Bruno; Meier, Peter J; Hofmann, Alan F; Sugiyama, Yuichi

2005-01-01

Bile salts are predominantly taken up by hepatocytes via the basolateral Na(+)-taurocholate cotransporting polypeptide (NTCP/SLC10A1) and secreted into the bile by the bile salt export pump (BSEP/ABCB11). In the present study, we transfected rat Ntcp and rat Bsep into polarized Madin-Darby canine kidney cells and characterized the transport properties of these cells for eight bile salts. Immunohistochemical staining demonstrated that Ntcp was expressed at the basolateral domains, whereas Bsep was expressed at the apical domains. Basal-to-apical transport of taurocholate across the monolayer expressing only Ntcp and that coexpressing Ntcp/Bsep was observed, whereas the flux across the monolayer of control and Bsep-expressing cells was symmetrical. Basal-to-apical transport of taurocholate across Ntcp/Bsep-coexpressing monolayers was significantly higher than that across monolayers expressing only Ntcp. Kinetic analysis of this vectorial transport of taurocholate gave an apparent K(m) value of 13.9 +/- 4.7 microM for cells expressing Ntcp alone, which is comparable with 22.2 +/- 4.5 microM for cells expressing both Ntcp and Bsep and V(max) values of 15.8 +/- 4.2 and 60.8 +/- 9.0 pmol.min(-1).mg protein(-1) for Ntcp alone and Ntcp and Bsep-coexpressing cells, respectively. Transcellular transport of cholate, glycocholate, taurochenodeoxycholate, chenodeoxycholate, glycochenodeoxycholate, tauroursodeoxycholate, ursodeoxycholate, and glycoursodeoxycholate, but not that of lithocholate was also observed across the double transfectant. This double-expressing system can be used as a model to clarify vectorial transport of bile salts across hepatocytes under physiological conditions.

First report of autochthonous non-vectorial canine leishmaniasis in New Caledonia, south-western Pacific: implications for new control measures and recommendations on importation of dogs.

PubMed

Daval, Nathalie; Marchal, Céline; Guillaumot, Laurent; Hüe, Thomas; Ravel, Christophe; Keck, Nicolas; Kasbari, Mohamed

2016-02-25

Canine leishmaniasis (CanL), a parasitic zoonotic disease caused by Leishmania infantum and usually transmitted by phlebotomine sandflies, has rarely been reported in Pacific islands, which have been regarded until now as leishmaniasis-free territory. Here, we report the first autochthonous CanL case in New Caledonia (south-western Pacific) and the investigations carried out 1) to determine how infection was introduced into and transmitted among these dogs and 2) to assess the risks to animal and public health. Extensive epidemiological and entomological investigations in and around the focus were carried out. Leishmaniasis infection was confirmed by histopathology, indirect fluorescent antibody test, real-time PCR, and culture. Parasite strain was typed by the isoenzymatic technique. The survey revealed close contacts between the autochthonous dog and two infected bitches imported from Spain, but failed to find any possible vector or disease spreading to other animals or humans. L. infantum zymodeme MON-1, the most frequent type in the Mediterranean basin, was identified. Although transplacental and venereal transmissions could not be excluded, the evidence was in favour of non-vectorial, direct dog-to-dog transmission. This study corroborates the possibility of non-vectorial routes (transplacental, venereal, and direct dog-to-dog) of canine leishmaniasis transmission in New Caledonia and raises the debate of relevant test requirements and diagnostic sensitivity prior to importation of dogs in Leishmania-free regions. New leishmaniasis control measures and recommendations to avoid future CanL introduction on the island are discussed.

Acute Lung Injury Edema Fluid Decreases Net Fluid Transport across Human Alveolar Epithelial Type II Cells*

PubMed Central

Lee, Jae W.; Fang, Xiaohui; Dolganov, Gregory; Fremont, Richard D.; Bastarache, Julie A.; Ware, Lorraine B.; Matthay, Michael A.

2009-01-01

Most patients with acute lung injury (ALI) have reduced alveolar fluid clearance that has been associated with higher mortality. Several mechanisms may contribute to the decrease in alveolar fluid clearance. In this study, we tested the hypothesis that pulmonary edema fluid from patients with ALI might reduce the expression of ion transport genes responsible for vectorial fluid transport in primary cultures of human alveolar epithelial type II cells. Following exposure to ALI pulmonary edema fluid, the gene copy number for the major sodium and chloride transport genes decreased. By Western blot analyses, protein levels of αENaC, α1Na,K-ATPase, and cystic fibrosis transmembrane conductance regulator decreased as well. In contrast, the gene copy number for several inflammatory cytokines increased markedly. Functional studies demonstrated that net vectorial fluid transport was reduced for human alveolar type II cells exposed to ALI pulmonary edema fluid compared with plasma (0.02±0.05 versus 1.31±0.56 μl/cm2/h, p<0.02). An inhibitor of p38 MAPK phosphorylation (SB202190) partially reversed the effects of the edema fluid on net fluid transport as well as gene and protein expression of the main ion transporters. In summary, alveolar edema fluid from patients with ALI induced a significant reduction in sodium and chloride transport genes and proteins in human alveolar epithelial type II cells, effects that were associated with a decrease in net vectorial fluid transport across human alveolar type II cell monolayers. PMID:17580309

Boric acid permeation in forward osmosis membrane processes: modeling, experiments, and implications.

PubMed

Jin, Xue; Tang, Chuyang Y; Gu, Yangshuo; She, Qianhong; Qi, Saren

2011-03-15

Forward osmosis (FO) is attracting increasing interest for its potential applications in desalination. In FO, permeation of contaminants from feed solution into draw solution through the semipermeable membrane can take place simultaneously with water diffusion. Understanding the contaminants transport through and rejection by FO membrane has significant technical implications in the way to separate clean water from the diluted draw solution. In this study, a model was developed to predict boron flux in FO operation. A strong agreement between modeling results and experimental data indicates that the model developed in this study can accurately predict the boron transport through FO membranes. Furthermore, the model can guide the fabrication of improved FO membranes with decreased boron permeability and structural parameter to minimize boron flux. Both theoretical model and experimental results demonstrated that when membrane active layer was facing draw solution, boron flux was substantially greater compared to the other membrane orientation due to more severe internal concentration polarization. In this investigation, for the first time, rejection of contaminants was defined in FO processes. This is critical to compare the membrane performance between different membranes and experimental conditions.

Proton Exchange Membrane Fuel Cell Engineering Model Powerplant. Test Report: Benchmark Tests in Three Spatial Orientations

NASA Technical Reports Server (NTRS)

Loyselle, Patricia; Prokopius, Kevin

2011-01-01

Proton exchange membrane (PEM) fuel cell technology is the leading candidate to replace the aging alkaline fuel cell technology, currently used on the Shuttle, for future space missions. This test effort marks the final phase of a 5-yr development program that began under the Second Generation Reusable Launch Vehicle (RLV) Program, transitioned into the Next Generation Launch Technologies (NGLT) Program, and continued under Constellation Systems in the Exploration Technology Development Program. Initially, the engineering model (EM) powerplant was evaluated with respect to its performance as compared to acceptance tests carried out at the manufacturer. This was to determine the sensitivity of the powerplant performance to changes in test environment. In addition, a series of tests were performed with the powerplant in the original standard orientation. This report details the continuing EM benchmark test results in three spatial orientations as well as extended duration testing in the mission profile test. The results from these tests verify the applicability of PEM fuel cells for future NASA missions. The specifics of these different tests are described in the following sections.

Lengthening of the Stargazin Cytoplasmic Tail Increases Synaptic Transmission by Promoting Interaction to Deeper Domains of PSD-95.

PubMed

Hafner, Anne-Sophie; Penn, Andrew C; Grillo-Bosch, Dolors; Retailleau, Natacha; Poujol, Christel; Philippat, Amandine; Coussen, Françoise; Sainlos, Matthieu; Opazo, Patricio; Choquet, Daniel

2015-04-22

PSD-95 is a prominent organizer of the postsynaptic density (PSD) that can present a filamentous orientation perpendicular to the plasma membrane. Interactions between PSD-95 and transmembrane proteins might be particularly sensitive to this orientation, as "long" cytoplasmic tails might be required to reach deeper PSD-95 domains. Extension/retraction of transmembrane protein C-tails offer a new way of regulating binding to PSD-95. Using stargazin as a model, we found that enhancing the apparent length of stargazin C-tail through phosphorylation or by an artificial linker was sufficient to potentiate binding to PSD-95, AMPAR anchoring, and synaptic transmission. A linear extension of stargazin C-tail facilitates binding to PSD-95 by preferentially engaging interaction with the farthest located PDZ domains regarding to the plasma membrane, which present a greater affinity for the stargazin PDZ-domain-binding motif. Our study reveals that the concerted orientation of the stargazin C-tail and PSD-95 is a major determinant of synaptic strength. Copyright © 2015 Elsevier Inc. All rights reserved.

Comparison of transmission parameters between Anopheles argyritarsis and Anopheles pseudopunctipennis in two ecologically different localities of Bolivia.

PubMed

Lardeux, Frédéric; Aliaga, Claudia; Tejerina, Rosenka; Torrez, Libia

2013-08-13

Anopheles (Anopheles) pseudopunctipennis is a recognized malaria vector in the slopes of the Andes of Bolivia. There, other species might be involved in malaria transmission and one candidate could be Anopheles argyritarsis. Although it is generally admitted that this species is not a malaria vector in the neotropical region, its potential role in transmission is still controversial and this situation has to be cleared, at least for Bolivia. Comparing the vectorial efficiency of An. pseudopunctipennis with that of An. argyritarsis could solve the question. The two species were sampled throughout Bolivia to estimate their degree of co-existence in their distribution range. Vectorial efficiencies of the two species were compared in two ecologically different localities where the species were sympatric by analysing their vectorial capacities and components (i e, human biting rates, human biting index, survival, durations of the gonotrophic cycle and extrinsic cycle), and the entomological inoculation rates (EIR). Mosquitoes were sampled monthly during more than one year in the two localities. A monthly sample consisted in hourly captures in four houses (inside and outside) in each locality, during four consecutive nights. Climatic variables (temperature, humidity, potential evapo-transpiration and precipitations) were recorded to better understand variability in the entomological parameters. Relationships were analysed using multivariate methods. Anopheles pseudopunctipennis and An. argyritarsis are "altitude" species, sharing the same geographical distribution range in the Andes of Bolivia. No Plasmodium parasite was identified in An. argyritarsis and estimates of the vectorial capacity indicated that it is not a malaria vector in the two studied localities, unlike An. pseudopunctipennis which showed positive EIRs. This latter species, although not a very good malaria vector, exhibited better life traits values and better behavioural characteristics in favour of transmission as compared to An. argyritarsis. In the Andes of Bolivia, above 1000 m of altitude, An. pseudopunctipennis is likely to be the only malaria vector. There, it is present almost everywhere and priority control effort should be directed toward this species. Below 1000 m of altitude, vector incrimination should also be focused on other sympatric species (likely not An. argyritarsis) that might be locally important. From the present study, candidates would be among Anopheles rangeli, Anopheles triannulatus s.l., Anopheles trinkae, Anopheles nuneztovari s.l., Anopheles oswaldoi s.l. and Anopheles benarrochi s.l.

Comparison of transmission parameters between Anopheles argyritarsis and Anopheles pseudopunctipennis in two ecologically different localities of Bolivia

PubMed Central

2013-01-01

Background Anopheles (Anopheles) pseudopunctipennis is a recognized malaria vector in the slopes of the Andes of Bolivia. There, other species might be involved in malaria transmission and one candidate could be Anopheles argyritarsis. Although it is generally admitted that this species is not a malaria vector in the neotropical region, its potential role in transmission is still controversial and this situation has to be cleared, at least for Bolivia. Comparing the vectorial efficiency of An. pseudopunctipennis with that of An. argyritarsis could solve the question. Methods The two species were sampled throughout Bolivia to estimate their degree of co-existence in their distribution range. Vectorial efficiencies of the two species were compared in two ecologically different localities where the species were sympatric by analysing their vectorial capacities and components (i e, human biting rates, human biting index, survival, durations of the gonotrophic cycle and extrinsic cycle), and the entomological inoculation rates (EIR). Mosquitoes were sampled monthly during more than one year in the two localities. A monthly sample consisted in hourly captures in four houses (inside and outside) in each locality, during four consecutive nights. Climatic variables (temperature, humidity, potential evapo-transpiration and precipitations) were recorded to better understand variability in the entomological parameters. Relationships were analysed using multivariate methods. Results Anopheles pseudopunctipennis and An. argyritarsis are “altitude” species, sharing the same geographical distribution range in the Andes of Bolivia. No Plasmodium parasite was identified in An. argyritarsis and estimates of the vectorial capacity indicated that it is not a malaria vector in the two studied localities, unlike An. pseudopunctipennis which showed positive EIRs. This latter species, although not a very good malaria vector, exhibited better life traits values and better behavioural characteristics in favour of transmission as compared to An. argyritarsis. Conclusions In the Andes of Bolivia, above 1000 m of altitude, An. pseudopunctipennis is likely to be the only malaria vector. There, it is present almost everywhere and priority control effort should be directed toward this species. Below 1000 m of altitude, vector incrimination should also be focused on other sympatric species (likely not An. argyritarsis) that might be locally important. From the present study, candidates would be among Anopheles rangeli, Anopheles triannulatus s.l., Anopheles trinkae, Anopheles nuneztovari s.l., Anopheles oswaldoi s.l. and Anopheles benarrochi s.l. PMID:23941216

Alteration of plant species assemblages can decrease the transmission potential of malaria mosquitoes.

PubMed

Ebrahimi, Babak; Jackson, Bryan T; Guseman, Julie L; Przybylowicz, Colin M; Stone, Christopher M; Foster, Woodbridge A

2018-03-01

Knowledge of the link between a vector population's pathogen-transmission potential and its biotic environment can generate more realistic forecasts of disease risk due to environmental change. It also can promote more effective vector control by both conventional and novel means.This study assessed the effect of particular plant species assemblages differing in nectar production on components of the vectorial capacity of the mosquito Anopheles gambiae s.s. , an important vector of African malaria.We followed cohorts of mosquitoes for three weeks in greenhouse mesocosms holding nectar-poor and nectar-rich plant species by tracking daily mortalities and estimating daily biting rates and fecundities. At death, a mosquito's insemination status and wing length were determined. These life history traits allowed incorporation of larval dynamics into a vectorial capacity estimate. This new study provided both novel assemblages of putative host plants and a human blood host within a nocturnal period of maximum biting.Survivorship was significantly greater in nectar-rich environments than nectar-poor ones, resulting in greater total fecundity. Daily biting rate and fecundity per female between treatments was not detected. These results translated to greater estimated vectorial capacities in the nectar-rich environment in all four replicates of the experiment (means: 1,089.5 ± 125.2 vs. 518.3 ± 60.6). When mosquito density was made a function of survival and fecundity, rather than held constant, the difference between plant treatments was more pronounced, but so was the variance, so differences were not statistically significant. In the nectar-poor environment, females' survival suffered severely when a blood host was not provided. A sugar-accessibility experiment confirmed that Parthenium hysterophorus is a nectar-poor plant for these mosquitoes. Synthesis and applications. This study, assessing the effect of particular plant species assemblages on the vectorial capacity of malaria mosquitoes, highlights the likelihood that changes in plant communities (e.g. due to introduction of exotic or nectar-rich species) can increase malaria transmission and that a reduction of favourable nectar sources can reduce it. Also, plant communities' data can be used to identify potential high risk areas. Further studies are warranted to explore how and when management of plant species assemblages should be considered as an option in an integrated vector management strategy.

How representative is the 'Representative Value' of refraction provided by the Shin-Nippon NVision-K 5001 autorefractor?

PubMed

Tang, Wing Chun; Tang, Ying Yung; Lam, Carly S Y

2014-01-01

The aim of the study was to evaluate the level of agreement between the 'Representative Value' (RV) of refraction obtained from the Shin-Nippon NVision-K 5001 instrument with values calculated from individual measurement readings using standard algebraic methods. Cycloplegic autorefraction readings for 101 myopic children aged 8-13 years (10.9 ± 1.42 years) were obtained using the Shin-Nippon NVision-K 5001. Ten autorefractor measurements were taken for each eye. The spherical equivalent (SE), sphere (Sph) and cylindrical component (Cyl) power of each eye were calculated, firstly, by averaging the 10 repeated measurements (Mean SE, Mean Sph and Mean Cyl), and secondly, by the vector representation method (Vector SE, Vector Sph and Vector Cyl). These calculated values were then compared with those of RV (RV SE, RV Sph and RV Cyl) provided by the proprietary software of the NVision-K 5001 using one-way analysis of variance (anova). The agreement between the methods was also assessed. The SE of the subjects ranged from -5.37 to -0.62 D (mean ± SD, = -2.89 ± 1.01 D). The Mean SE was in exact agreement with the Vector SE. There were no significant differences between the RV readings and those calculated using non-vectorial or vectorial methods for any of the refractive powers (SE, p = 0.99; Sph, p = 0.93; Cyl, p = 0.24). The (mean ± SD) differences were: RV SE vs Mean SE (and also RV SE vs Vector SE) -0.01 ± 0.06 D; RV Sph vs Mean Sph, -0.01 ± 0.05 D; RV Sph vs Vector Sph, -0.04 ± 0.06 D; RV Cyl vs Mean Cyl, 0.01 ± 0.07 D; RV Cyl vs Vector Cyl, 0.06 ± 0.09 D. Ninety-eight percent of RV reading differed from their non-vectorial or vectorial counterparts by less than 0.25 D. The RV values showed good agreement to the results calculated using conventional methods. Although the formula used to calculate RV by the NVision-K 5001 autorefractor is proprietary, our results provide validation for the use of RV measurements in clinical practice and vision science research. © 2013 The Authors Ophthalmic & Physiological Optics © 2013 The College of Optometrists.

Elasticity of smectic liquid crystals with in-plane orientational order and dispiration asymmetry

NASA Astrophysics Data System (ADS)

Alageshan, Jaya Kumar; Chakrabarti, Buddhapriya; Hatwalne, Yashodhan

2017-02-01

The Nelson-Peliti formulation of the elasticity theory of isolated fluid membranes with orientational order emphasizes the interplay between geometry, topology, and thermal fluctuations. Fluid layers of lamellar liquid crystals such as smectic-C , hexatic smectics, and smectic-C* are endowed with in-plane orientational order. We extend the Nelson-Peliti formulation so as to bring these smectics within its ambit. Using the elasticity theory of smectics-C*, we show that positive and negative dispirations (topological defects in Smectic-C* liquid crystals) with strengths of equal magnitude have disparate energies—a result that is amenable to experimental tests.

Structure refinement of membrane proteins via molecular dynamics simulations.

PubMed

Dutagaci, Bercem; Heo, Lim; Feig, Michael

2018-07-01

A refinement protocol based on physics-based techniques established for water soluble proteins is tested for membrane protein structures. Initial structures were generated by homology modeling and sampled via molecular dynamics simulations in explicit lipid bilayer and aqueous solvent systems. Snapshots from the simulations were selected based on scoring with either knowledge-based or implicit membrane-based scoring functions and averaged to obtain refined models. The protocol resulted in consistent and significant refinement of the membrane protein structures similar to the performance of refinement methods for soluble proteins. Refinement success was similar between sampling in the presence of lipid bilayers and aqueous solvent but the presence of lipid bilayers may benefit the improvement of lipid-facing residues. Scoring with knowledge-based functions (DFIRE and RWplus) was found to be as good as scoring using implicit membrane-based scoring functions suggesting that differences in internal packing is more important than orientations relative to the membrane during the refinement of membrane protein homology models. © 2018 Wiley Periodicals, Inc.

Structure and hydration of membranes embedded with voltage-sensing domains.

PubMed

Krepkiy, Dmitriy; Mihailescu, Mihaela; Freites, J Alfredo; Schow, Eric V; Worcester, David L; Gawrisch, Klaus; Tobias, Douglas J; White, Stephen H; Swartz, Kenton J

2009-11-26

Despite the growing number of atomic-resolution membrane protein structures, direct structural information about proteins in their native membrane environment is scarce. This problem is particularly relevant in the case of the highly charged S1-S4 voltage-sensing domains responsible for nerve impulses, where interactions with the lipid bilayer are critical for the function of voltage-activated ion channels. Here we use neutron diffraction, solid-state nuclear magnetic resonance (NMR) spectroscopy and molecular dynamics simulations to investigate the structure and hydration of bilayer membranes containing S1-S4 voltage-sensing domains. Our results show that voltage sensors adopt transmembrane orientations and cause a modest reshaping of the surrounding lipid bilayer, and that water molecules intimately interact with the protein within the membrane. These structural findings indicate that voltage sensors have evolved to interact with the lipid membrane while keeping energetic and structural perturbations to a minimum, and that water penetrates the membrane, to hydrate charged residues and shape the transmembrane electric field.

Structure and dynamics of cationic membrane peptides and proteins: Insights from solid-state NMR

PubMed Central

Hong, Mei; Su, Yongchao

2011-01-01

Many membrane peptides and protein domains contain functionally important cationic Arg and Lys residues, whose insertion into the hydrophobic interior of the lipid bilayer encounters significant energy barriers. To understand how these cationic molecules overcome the free energy barrier to insert into the lipid membrane, we have used solid-state NMR spectroscopy to determine the membrane-bound topology of these peptides. A versatile array of solid-state NMR experiments now readily yields the conformation, dynamics, orientation, depth of insertion, and site-specific protein–lipid interactions of these molecules. We summarize key findings of several Arg-rich membrane peptides, including β-sheet antimicrobial peptides, unstructured cell-penetrating peptides, and the voltage-sensing helix of voltage-gated potassium channels. Our results indicate the central role of guanidinium-phosphate and guanidinium-water interactions in dictating the structural topology of these cationic molecules in the lipid membrane, which in turn account for the mechanisms of this functionally diverse class of membrane peptides. PMID:21344534

Structure and hydration of membranes embedded with voltage-sensing domains

PubMed Central

Krepkiy, Dmitriy; Mihailescu, Mihaela; Freites, J. Alfredo; Schow, Eric V.; Worcester, David L.; Gawrisch, Klaus; Tobias, Douglas; White, Stephen H.; Swartz, Kenton J.

2009-01-01

Despite the growing number of atomic-resolution membrane protein structures, direct structural information about proteins in their native membrane environment is scarce. This problem is particularly relevant in the case of the highly-charged S1–S4 voltage-sensing domains responsible for nerve impulses, where interactions with the lipid bilayer are critical for the function of voltage-activated potassium channels. Here we use neutron diffraction, solid-state nuclear magnetic resonance spectroscopy, and molecular dynamics simulations to investigate the structure and hydration of bilayer membranes containing S1–S4 voltage-sensing domains. Our results show that voltage sensors adopt transmembrane orientations, cause a modest reshaping of the surrounding lipid bilayer, and that water molecules intimately interact with the protein within the membrane. These structural findings reveal that voltage sensors have evolved to interact with the lipid membrane while keeping the energetic and structural perturbations to a minimum, and that water penetrates into the membrane to hydrate charged residues and shape the transmembrane electric field. PMID:19940918

PEM/SPE fuel cell

DOEpatents

Grot, S.A.

1998-01-13

A PEM/SPE fuel cell is described including a membrane-electrode assembly (MEA) having a plurality of oriented filament embedded the face thereof for supporting the MEA and conducting current therefrom to contiguous electrode plates. 4 figs.

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

Kardash, Maria E.; Dzuba, Sergei A., E-mail: dzuba@kinetics.nsc.ru

Lipid-cholesterol interactions are responsible for different properties of biological membranes including those determining formation in the membrane of spatial inhomogeneities (lipid rafts). To get new information on these interactions, electron spin echo (ESE) spectroscopy, which is a pulsed version of electron paramagnetic resonance (EPR), was applied to study 3β-doxyl-5α-cholestane (DCh), a spin-labeled analog of cholesterol, in phospholipid bilayer consisted of equimolecular mixture of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine and 1,2-dioleoyl-sn-glycero-3-phosphocholine. DCh concentration in the bilayer was between 0.1 mol.% and 4 mol.%. For comparison, a reference system containing a spin-labeled 5-doxyl-stearic acid (5-DSA) instead of DCh was studied as well. The effects of “instantaneousmore » diffusion” in ESE decay and in echo-detected (ED) EPR spectra were explored for both systems. The reference system showed good agreement with the theoretical prediction for the model of spin labels of randomly distributed orientations, but the DCh system demonstrated remarkably smaller effects. The results were explained by assuming that neighboring DCh molecules are oriented in a correlative way. However, this correlation does not imply the formation of clusters of cholesterol molecules, because conventional continuous wave EPR spectra did not show the typical broadening due to aggregation of spin labels and the observed ESE decay was not faster than in the reference system. So the obtained data evidence that cholesterol molecules at low concentrations in biological membranes can interact via large distances of several nanometers which results in their orientational self-ordering.« less

Orientation and cellular distribution of membrane-bound catechol-O-methyltransferase in cortical neurons: implications for drug development.

PubMed

Chen, Jingshan; Song, Jian; Yuan, Peixiong; Tian, Qingjun; Ji, Yuanyuan; Ren-Patterson, Renee; Liu, Guangping; Sei, Yoshitasu; Weinberger, Daniel R

2011-10-07

Catechol-O-methyltransferase (COMT) is a key enzyme for inactivation and metabolism of catechols, including dopamine, norepinephrine, caffeine, and estrogens. It plays an important role in cognition, arousal, pain sensitivity, and stress reactivity in humans and in animal models. The human COMT gene is associated with a diverse spectrum of human behaviors and diseases from cognition and psychiatric disorders to chronic pain and cancer. There are two major forms of COMT proteins, membrane-bound (MB) COMT and soluble (S) COMT. MB-COMT is the main form in the brain. The cellular distribution of MB-COMT in cortical neurons remains unclear and the orientation of MB-COMT on the cellular membrane is controversial. In this study, we demonstrate that MB-COMT is located in the cell body and in axons and dendrites of rat cortical neurons. Analyses of MB-COMT orientation with computer simulation, flow cytometry and a cell surface enzyme assay reveal that the C-terminal catalytic domain of MB-COMT is in the extracellular space, which suggests that MB-COMT can inactivate synaptic and extrasynaptic dopamine on the surface of presynaptic and postsynaptic neurons. Finally, we show that the COMT inhibitor tolcapone induces cell death via the mechanism of apoptosis, and its cytotoxicity is dependent on dosage and correlated with COMT Val/Met genotypes in human lymphoblastoid cells. These results suggest that MB-COMT specific inhibitors can be developed and that tolcapone may be less hazardous at low doses and in specific genetic backgrounds.

Rotational behaviour of PEGylated gold nanorods in a lipid bilayer system

NASA Astrophysics Data System (ADS)

Oroskar, Priyanka A.; Jameson, Cynthia J.; Murad, Sohail

2017-06-01

PEGylated gold nanorods are widely used as nanocarriers in targeted drug delivery and other nanotechnology applications due to the special optical and photo-thermal characteristics of gold nanorods. In this work, we employ coarse-grain molecular simulations to examine the pathway by which PEGylated gold nanorods enter and exit a dipalmitoylphosphatidylcholine lipid bilayer membrane and follow the behaviour of the system to investigate the consequences. We find that PEGylated gold nanorods rotate during permeation, lying down and straightening up as they make their way through the lipid membrane. We find that this rotational behaviour, irrespective of the initial orientation of the nanorod with respect to the membrane normal, is concomitant with the changing interactions of polyethylene glycol (PEG) beads with lipid head beads in both membrane leaflets. For a nanorod with hydrophilic ligands, such as PEG, lying down appears to be driven by favourable hydrophilic interactions with the phosphate and choline groups of the lipid. Mobility of the ligands offers mechanisms for these favourable interactions and for minimising unfavourable interactions with the hydrophobic lipid tails that constitute the inner section of the membrane; the PEG ligands can stretch out to reach the phosphate and choline groups of both leaflets and they can coil in and interact with each other and avoid the alkane lipid tails. Recently developed experimental techniques for imaging, orientation, and rotation of single gold nanorods may be able to observe this predicted rotational behaviour. We find that lipid flip-flop mechanisms do not differ significantly from a spherical gold nanoparticle to a gold nanorod, and PEGylated gold nanorods like their spherical counterparts do not remove lipid molecules from the bilayer membrane. Our results should be of interest to experimentalists who plan to use functionalised gold nanorods in biomedical applications.

Fatty acid fouling of forward osmosis membrane: Effects of pH, calcium, membrane orientation, initial permeate flux and foulant composition.

PubMed

Zhao, Pin; Gao, Baoyu; Yue, Qinyan; Liu, Pan; Shon, Ho Kyong

2016-08-01

Octanoic acid (OA) was selected to represent fatty acids in effluent organic matter (EOM). The effects of feed solution (FS) properties, membrane orientation and initial permeate flux on OA fouling in forward osmosis (FO) were investigated. The undissociated OA formed a cake layer quickly and caused the water flux to decline significantly in the initial 0.5hr at unadjusted pH3.56; while the fully dissociated OA behaved as an anionic surfactant and promoted the water permeation at an elevated pH of 9.00. Moreover, except at the initial stage, the sudden decline of water flux (meaning the occurrence of severe membrane fouling) occurred in two conditions: 1. 0.5mmol/L Ca(2+), active layer facing draw solution (AL-DS) and 1.5mol/L NaCl (DS); 2. No Ca(2+), active layer-facing FS (AL-FS) and 4mol/L NaCl (DS). This demonstrated that cake layer compaction or pore blocking occurred only when enough foulants were absorbed into the membrane surface, and the water permeation was high enough to compact the deposit inside the porous substrate. Furthermore, bovine serum albumin (BSA) was selected as a co-foulant. The water flux of both co-foulants was between the fluxes obtained separately for the two foulants at pH3.56, and larger than the two values at pH9.00. This manifested that, at pH3.56, BSA alleviated the effect of the cake layer caused by OA, and OA enhanced BSA fouling simultaneously; while at pH9.00, the mutual effects of OA and BSA eased the membrane fouling. Copyright © 2016. Published by Elsevier B.V.

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

PubMed

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.

Secretion of the Intimin Passenger Domain Is Driven by Protein Folding*

PubMed Central

Leo, Jack C.; Oberhettinger, Philipp; Yoshimoto, Shogo; Udatha, D. B. R. K. Gupta; Morth, J. Preben; Schütz, Monika; Hori, Katsutoshi

2016-01-01

Intimin is an essential adhesin of attaching and effacing organisms such as entropathogenic Escherichia coli. It is also the prototype of type Ve secretion or inverse autotransport, where the extracellular C-terminal region or passenger is exported with the help of an N-terminal transmembrane β-barrel domain. We recently reported a stalled secretion intermediate of intimin, where the passenger is located in the periplasm but the β-barrel is already inserted into the membrane. Stalling of this mutant is due to the insertion of an epitope tag at the very N terminus of the passenger. Here, we examined how this insertion disrupts autotransport and found that it causes misfolding of the N-terminal immunoglobulin (Ig)-like domain D00. We could also stall the secretion by making an internal deletion in D00, and introducing the epitope tag into the second Ig-like domain, D0, also resulted in reduced passenger secretion. In contrast to many classical autotransporters, where a proximal folding core in the passenger is required for secretion, the D00 domain is dispensable, as the passenger of an intimin mutant lacking D00 entirely is efficiently exported. Furthermore, the D00 domain is slightly less stable than the D0 and D1 domains, unfolding at ∼200 piconewtons (pN) compared with ∼250 pN for D0 and D1 domains as measured by atomic force microscopy. Our results support a model where the secretion of the passenger is driven by sequential folding of the extracellular Ig-like domains, leading to vectorial transport of the passenger domain across the outer membrane in an N to C direction. PMID:27466361

A wrinkle in flight: the role of elastin fibres in the mechanical behaviour of bat wing membranes

PubMed Central

Cheney, Jorn A.; Konow, Nicolai; Bearnot, Andrew; Swartz, Sharon M.

2015-01-01

Bats fly using a thin wing membrane composed of compliant, anisotropic skin. Wing membrane skin deforms dramatically as bats fly, and its three-dimensional configurations depend, in large part, on the mechanical behaviour of the tissue. Large, macroscopic elastin fibres are an unusual mechanical element found in the skin of bat wings. We characterize the fibre orientation and demonstrate that elastin fibres are responsible for the distinctive wrinkles in the surrounding membrane matrix. Uniaxial mechanical testing of the wing membrane, both parallel and perpendicular to elastin fibres, is used to distinguish the contribution of elastin and the surrounding matrix to the overall membrane mechanical behaviour. We find that the matrix is isotropic within the plane of the membrane and responsible for bearing load at high stress; elastin fibres are responsible for membrane anisotropy and only contribute substantially to load bearing at very low stress. The architecture of elastin fibres provides the extreme extensibility and self-folding/self-packing of the wing membrane skin. We relate these findings to flight with membrane wings and discuss the aeromechanical significance of elastin fibre pre-stress, membrane excess length, and how these parameters may aid bats in resisting gusts and preventing membrane flutter. PMID:25833238

Membrane-Induced Structural Rearrangement and Identification of a Novel Membrane Anchor in Talin F2F3

PubMed Central

Arcario, Mark J.; Tajkhorshid, Emad

2014-01-01

Experimental challenges associated with characterization of the membrane-bound form of talin have prevented us from understanding the molecular mechanism of its membrane-dependent integrin activation. Here, utilizing what we believe to be a novel membrane mimetic model, we present a reproducible model of membrane-bound talin observed across multiple independent simulations. We characterize both local and global membrane-induced structural transitions that successfully reconcile discrepancies between biochemical and structural studies and provide insight into how talin might modulate integrin function. Membrane binding of talin, captured in unbiased simulations, proceeds through three distinct steps: initial electrostatic recruitment of the F2 subdomain to anionic lipids via several basic residues; insertion of an initially buried, conserved hydrophobic anchor into the membrane; and association of the F3 subdomain with the membrane surface through a large, interdomain conformational change. These latter two steps, to our knowledge, have not been observed or described previously. Electrostatic analysis shows talin F2F3 to be highly polarized, with a highly positive underside, which we attribute to the initial electrostatic recruitment, and a negative top face, which can help orient the protein optimally with respect to the membrane, thereby reducing the number of unproductive membrane collision events. PMID:25418091

A wrinkle in flight: the role of elastin fibres in the mechanical behaviour of bat wing membranes.

PubMed

Cheney, Jorn A; Konow, Nicolai; Bearnot, Andrew; Swartz, Sharon M

2015-05-06

Bats fly using a thin wing membrane composed of compliant, anisotropic skin. Wing membrane skin deforms dramatically as bats fly, and its three-dimensional configurations depend, in large part, on the mechanical behaviour of the tissue. Large, macroscopic elastin fibres are an unusual mechanical element found in the skin of bat wings. We characterize the fibre orientation and demonstrate that elastin fibres are responsible for the distinctive wrinkles in the surrounding membrane matrix. Uniaxial mechanical testing of the wing membrane, both parallel and perpendicular to elastin fibres, is used to distinguish the contribution of elastin and the surrounding matrix to the overall membrane mechanical behaviour. We find that the matrix is isotropic within the plane of the membrane and responsible for bearing load at high stress; elastin fibres are responsible for membrane anisotropy and only contribute substantially to load bearing at very low stress. The architecture of elastin fibres provides the extreme extensibility and self-folding/self-packing of the wing membrane skin. We relate these findings to flight with membrane wings and discuss the aeromechanical significance of elastin fibre pre-stress, membrane excess length, and how these parameters may aid bats in resisting gusts and preventing membrane flutter. © 2015 The Author(s) Published by the Royal Society. All rights reserved.

Preparation, Processing, and Characterization of Oriented Polycrystalline Zeolite and Aluminophosphate Membranes

NASA Astrophysics Data System (ADS)

Stoeger, Jared Andrew

Since the advent of zeolite membranes, speculation on their industrial applicability has been closely monitored, although widespread commercialization has been hampered by limitations in fabrication and post-synthesis processing. Economical, energy-efficient technology breakthroughs require an evaluation of a range of material candidates which show robustness and reliability. Straightforward manufacturing techniques should be devised to generate thousands of square meters of membrane area; however, this demands control of structural characteristics on the scale of nanometers. As described in this dissertation, the path forward will be forged by exploiting the intrinsic crystalline properties of zeolites or aluminophosphates for the next advancement in membrane technology. A facile method is described for the preparation of silicalite-1 (MFI zeolite type) membranes using the secondary growth technique on symmetric porous stainless steel tubes. Activation through rapid thermal processing (RTP), a lamp-based heat-treatment process used as a critical fabrication step in silicon integrated circuit manufacturing, is proven to reduce the density of non-zeolitic transport pathways which are detrimental to high-resolution molecular sieving. RTP-treated membranes are shown to have enhanced performance in the binary separation of vapor-phase isomers (p-/o-xylene), gas-phase isomers (n-/i-butane), and alcohol/water when compared to membranes activated at a much slower heating rate but otherwise similarly-prepared. The performance is discussed in the context of the market potential for industrially-attractive separations: the recovery of p-xylene from an isomeric mixture or alcohol biofuels from aqueous post-fermentation streams. Hydrothermal growth techniques for the preparation and characterization of continuous aluminophosphate (AFI zeolite type) membranes with a preferential crystallographic alignment on porous alpha-Al2O3 disc supports are demonstrated. A mechanism is proposed for flake-like crystal formation in the early stages of in-plane crystalline intergrowth between oriented columnar crystals by electric heating. It is shown that elevated temperatures induce a phase transformation to a densified aluminophosphate phase despite framework metal substitution or alternative heat-treatment conditions. Additionally, stability and membrane characteristics following in situ microwave growth using a TiO2-coated support are examined. Indications of improved quality validate the candidacy of the microwave-grown membranes with regard to the potential for carbon nanotube synthesis in the aligned one-dimensional channels for high flux, high separation factor membrane fabrication.

Characterization of the Structure and Membrane Interaction of the Antimicrobial Peptides Aurein 2.2 and 2.3 from Australian Southern Bell Frogs

PubMed Central

Pan, Yeang-Ling; Cheng, John T.-J.; Hale, John; Pan, Jinhe; Hancock, Robert E. W.; Straus, Suzana K.

2007-01-01

The structure and membrane interaction of the antimicrobial peptide aurein 2.2 (GLFDIVKKVVGALGSL-CONH2), aurein 2.3 (GLFDIVKKVVGAIGSL-CONH2), both from Litoria aurea, and a carboxy C-terminal analog of aurein 2.3 (GLFDIVKKVVGAIGSL-COOH) were studied to determine which features of this class of peptides are key to activity. Circular dichroism and solution-state NMR data indicate that all three peptides adopt an α-helical structure in the presence of trifluoroethanol or lipids such as 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and a 1:1 mixture of DMPC and 1,2-dimyristoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DMPG). Oriented circular dichroism was used to determine the orientation of the peptides in lipid bilayers over a range of concentrations (peptide/lipid molar ratios (P/L) = 1:15–1:120) in DMPC and 1:1 DMPC/DMPG, in the liquid crystalline state. The results demonstrate that in DMPC all three peptides are surface adsorbed over a range of low peptide concentrations but insert into the bilayers at high peptide concentrations. This finding is corroborated by 31P-solid-state NMR data of the three peptides in DMPC, which shows that at high peptide concentrations the peptides perturb the membrane. Oriented circular dichroism data of the aurein peptides in 1:1 DMPC/DMPG, on the other hand, show that the peptides with amidated C-termini readily insert into the membrane bilayers over the concentration range studied (P/L = 1:15–1:120), whereas the aurein 2.3 peptide with a carboxy C-terminus inserts at a threshold concentration of P/L* between 1:80 and 1:120. Overall, the data presented here suggest that all three peptides studied interact with phosphatidylcholine membranes in a manner which is similar to aurein 1.2 and citropin 1.1, as reported in the literature, with no correlation to the reported activity. On the other hand, both aurein 2.2 and aurein 2.3 behave similarly in phosphatidylcholine/phosphatidylglycerol (PC/PG) membranes, whereas aurein 2.3-COOH inserts less readily. As this does not correlate with reported activities, minimal inhibitory concentrations of the three peptides against Staphylococcus aureus (strain C622, ATCC 25923) and Staphylococcus epidermidis (strain C621—clinical isolate) were determined. The correlation between structure, membrane interaction, and activity are discussed in light of these results. PMID:17259271

Characterization of the structure and membrane interaction of the antimicrobial peptides aurein 2.2 and 2.3 from Australian southern bell frogs.

PubMed

Pan, Yeang-Ling; Cheng, John T-J; Hale, John; Pan, Jinhe; Hancock, Robert E W; Straus, Suzana K

2007-04-15

The structure and membrane interaction of the antimicrobial peptide aurein 2.2 (GLFDIVKKVVGALGSL-CONH(2)), aurein 2.3 (GLFDIVKKVVGAIGSL-CONH(2)), both from Litoria aurea, and a carboxy C-terminal analog of aurein 2.3 (GLFDIVKKVVGAIGSL-COOH) were studied to determine which features of this class of peptides are key to activity. Circular dichroism and solution-state NMR data indicate that all three peptides adopt an alpha-helical structure in the presence of trifluoroethanol or lipids such as 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) and a 1:1 mixture of DMPC and 1,2-dimyristoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] (DMPG). Oriented circular dichroism was used to determine the orientation of the peptides in lipid bilayers over a range of concentrations (peptide/lipid molar ratios (P/L) = 1:15-1:120) in DMPC and 1:1 DMPC/DMPG, in the liquid crystalline state. The results demonstrate that in DMPC all three peptides are surface adsorbed over a range of low peptide concentrations but insert into the bilayers at high peptide concentrations. This finding is corroborated by (31)P-solid-state NMR data of the three peptides in DMPC, which shows that at high peptide concentrations the peptides perturb the membrane. Oriented circular dichroism data of the aurein peptides in 1:1 DMPC/DMPG, on the other hand, show that the peptides with amidated C-termini readily insert into the membrane bilayers over the concentration range studied (P/L = 1:15-1:120), whereas the aurein 2.3 peptide with a carboxy C-terminus inserts at a threshold concentration of P/L* between 1:80 and 1:120. Overall, the data presented here suggest that all three peptides studied interact with phosphatidylcholine membranes in a manner which is similar to aurein 1.2 and citropin 1.1, as reported in the literature, with no correlation to the reported activity. On the other hand, both aurein 2.2 and aurein 2.3 behave similarly in phosphatidylcholine/phosphatidylglycerol (PC/PG) membranes, whereas aurein 2.3-COOH inserts less readily. As this does not correlate with reported activities, minimal inhibitory concentrations of the three peptides against Staphylococcus aureus (strain C622, ATCC 25923) and Staphylococcus epidermidis (strain C621--clinical isolate) were determined. The correlation between structure, membrane interaction, and activity are discussed in light of these results.

Effect of Sterilization Methods on Electrospun Poly(lactic acid) (PLA) Fiber Alignment for Biomedical Applications.

PubMed

Valente, T A M; Silva, D M; Gomes, P S; Fernandes, M H; Santos, J D; Sencadas, V

2016-02-10

Medically approved sterility methods should be a major concern when developing a polymeric scaffold, mainly when commercialization is envisaged. In the present work, poly(lactic acid) (PLA) fiber membranes were processed by electrospinning with random and aligned fiber alignment and sterilized under UV, ethylene oxide (EO), and γ-radiation, the most common ones for clinical applications. It was observed that UV light and γ-radiation do not influence fiber morphology or alignment, while electrospun samples treated with EO lead to fiber orientation loss and morphology changing from cylindrical fibers to ribbon-like structures, accompanied to an increase of polymer crystallinity up to 28%. UV light and γ-radiation sterilization methods showed to be less harmful to polymer morphology, without significant changes in polymer thermal and mechanical properties, but a slight increase of polymer wettability was detected, especially for the samples treated with UV radiation. In vitro results indicate that both UV and γ-radiation treatments of PLA membranes allow the adhesion and proliferation of MG 63 osteoblastic cells in a close interaction with the fiber meshes and with a growth pattern highly sensitive to the underlying random or aligned fiber orientation. These results are suggestive of the potential of both γ-radiation sterilized PLA membranes for clinical applications in regenerative medicine, especially those where customized membrane morphology and fiber alignment is an important issue.

Orientation and Interaction of Oblique Cylindrical Inclusions Embedded in a Lipid Monolayer: A Theoretical Model for Viral Fusion Peptides

PubMed Central

Kozlovsky, Yonathan; Zimmerberg, Joshua; Kozlov, Michael M.

2004-01-01

We consider the elastic behavior of flat lipid monolayer embedding cylindrical inclusions oriented obliquely with respect to the monolayer plane. An oblique inclusion models a fusion peptide, a part of a specialized protein capable of inducing merger of biological membranes in the course of fundamental cellular processes. Although the crucial importance of the fusion peptides for membrane merger is well established, the molecular mechanism of their action remains unknown. This analysis is aimed at revealing mechanical deformations and stresses of lipid monolayers induced by the fusion peptides, which, potentially, can destabilize the monolayer structure and enhance membrane fusion. We calculate the deformation of a monolayer embedding a single oblique inclusion and subject to a lateral tension. We analyze the membrane-mediated interactions between two inclusions, taking into account bending of the monolayer and tilt of the hydrocarbon chains with respect to the surface normal. In contrast to a straightforward prediction that the oblique inclusions should induce tilt of the lipid chains, our analysis shows that the monolayer accommodates the oblique inclusion solely by bending. We find that the interaction between two inclusions varies nonmonotonically with the interinclusion distance and decays at large separations as square of the distance, similar to the electrostatic interaction between two electric dipoles in two dimensions. This long-range interaction is predicted to dominate the other interactions previously considered in the literature. PMID:15298906

Fluorine-Containing novel spatial and contact repellents

USDA-ARS?s Scientific Manuscript database

Mosquito-transmitted diseases such as malaria, dengue, and yellow fever, result in thousands of human deaths yearly. Climate change and global warming can enhance the vectorial capacity, and the temporal and spatial distribution of mosquito populations. To find more effective tools for mosquito and ...

Role of Transmembrane Potential and Defects on the Permeabilization of Lipid Bilayers by Alamethicin, an Ion-Channel-Forming Peptide.

PubMed

Su, ZhangFei; Shodiev, Muzaffar; Leitch, J Jay; Abbasi, Fatemeh; Lipkowski, Jacek

2018-05-29

The insertion and ion-conducting channel properties of alamethicin reconstituted into a 1,2-di- O-phytanyl- sn-glycero-3-phosphocholine bilayer floating on the surface of a gold (111) electrode modified with a 1-thio-β-d-glucose (β-Tg) self-assembled monolayer were investigated using a combination of electrochemical impedance spectroscopy (EIS) and polarization modulation infrared reflection absorption spectroscopy (PM-IRRAS). The hydrophilic β-Tg monolayer separated the bilayer from the gold substrate and created a water-rich spacer region, which better represents natural cell membranes. The EIS measurements acquired information about the membrane resistivity (a measure of membrane porosity), and the PM-IRRAS experiments provided insight into the conformation and orientation of the membrane constituents as a function of the transmembrane potential. The results showed that the presence of alamethicin had a small effect on the conformation and orientation of phospholipid molecules within the bilayer for all studied potentials. In contrast, the alamethicin peptides assumed a surface state, where the helical axes adopted a large tilt angle with respect to the surface normal, at small transmembrane potentials, and inserted into the bilayer at sufficiently negative transmembrane potentials forming pores, which behaved as barrel-stave ion channels for ionic transport across the membrane. The results indicated that insertion of alamethincin peptides into the bilayer was driven by the dipole-field interactions and that the transitions between the inserted and surface states were electrochemically reversible. Additionally, the EIS measurements performed on phospholipid bilayers without alamethicin also showed that the application of negative transmembrane potentials introduces defects into the bilayer. The membrane resistances measured in both the absence and presence of alamethicin show similar dependencies on the electrode potential, suggesting that the insertion of the peptide may also be assisted by the electroporation of the membrane. The findings in this study provide new insights into the mechanism of alamethicin insertion into phospholipid bilayers.

Effects of Lys to Glu mutations in GsMTx4 on membrane binding, peptide orientation, and self-association propensity, as analyzed by molecular dynamics simulations.

PubMed

Nishizawa, Kazuhisa; Nishizawa, Manami; Gnanasambandam, Radhakrishnan; Sachs, Frederick; Sukharev, Sergei I; Suchyna, Thomas M

2015-11-01

GsMTx4, a gating modifier peptide acting on cationic mechanosensitive channels, has a positive charge (+5e) due to six Lys residues. The peptide does not have a stereospecific binding site on the channel but acts from the boundary lipids within a Debye length of the pore probably by changing local stress. To gain insight into how these Lys residues interact with membranes, we performed molecular dynamics simulations of Lys to Glu mutants in parallel with our experimental work. In silico, K15E had higher affinity for 1-palmitoyl-2-oleoyl-glycero-3-phosphocholine bilayers than wild-type (WT) peptide or any other mutant tested, and showed deeper penetration than WT, a finding consistent with the experimental data. Experimentally, the inhibitory activities of K15E and K25E were most compromised, whereas K8E and K28E inhibitory activities remained similar to WT peptide. Binding of WT in an interfacial mode did not influence membrane thickness. With interfacial binding, the direction of the dipole moments of K15E and K25E was predicted to differ from WT, whereas those of K8E and K28E oriented similarly to that of WT. These results support a model in which binding of GsMTx4 to the membrane acts like an immersible wedge that serves as a membrane expansion buffer reducing local stress and thus inhibiting channel activity. In simulations, membrane-bound WT attracted other WT peptides to form aggregates. This may account for the positive cooperativity observed in the ion channel experiments. The Lys residues seem to fine-tune the depth of membrane binding, the tilt angle, and the dipole moments. Copyright © 2015 Elsevier B.V. All rights reserved.

Molecular Dynamics of a Water-Lipid Bilayer Interface

NASA Technical Reports Server (NTRS)

Wilson, Michael A.; Pohorille, Andrew

1994-01-01

We present results of molecular dynamics simulations of a glycerol 1-monooleate bilayer in water. The total length of analyzed trajectories is 5ns. The calculated width of the bilayer agrees well with the experimentally measured value. The interior of the membrane is in a highly disordered fluid state. Atomic density profile, orientational and conformational distribution functions, and order parameters indicate that disorder increases toward the center of the bilayer. Analysis of out-of-plane thermal fluctuations of the bilayer surfaces occurring at the time scale of the present calculations reveals that the distribution of modes agrees with predictions of the capillary wave model. Fluctuations of both bilayer surfaces are uncorrelated, yielding Gaussian distribution of instantaneous widths of the membrane. Fluctuations of the width produce transient thinning defects in the bilayer which occasionally span almost half of the membrane. The leading mechanism of these fluctuations is the orientational and conformational motion of head groups rather than vertical motion of the whole molecules. Water considerably penetrates the head group region of the bilayer but not its hydrocarbon core. The total net excess dipole moment of the interfacial water points toward the aqueous phase, but the water polarization profile is non-monotonic. Both water and head groups significantly contribute to the surface potential across the interface. The calculated sign of the surface potential is in agreement with that from experimental measurements, but the value is markedly overestimated. The structural and electrical properties of the water-bilayer system are discussed in relation to membrane functions, in particular transport of ions and nonelectrolytes across membranes.

Biocontrol of Listeria monocytogenes and Escherichia coli O157:H7 in Meat by Using Phages Immobilized on Modified Cellulose Membranes ▿

PubMed Central

Anany, H.; Chen, W.; Pelton, R.; Griffiths, M. W.

2011-01-01

The ability of phages to specifically interact with and lyse their host bacteria makes them ideal antibacterial agents. The range of applications of bacteriophage can be extended by their immobilization on inert surfaces. A novel method for the oriented immobilization of bacteriophage has been developed. The method was based on charge differences between the bacteriophage head, which exhibits an overall net negative charge, and the tail fibers, which possess an overall net positive charge. Hence, the head would be more likely to attach to positively charged surfaces, leaving the tails free to capture and lyse bacteria. Cellulose membranes modified so that they had a positive surface charge were used as the support for phage immobilization. It was established that the number of infective phages immobilized on the positively charged cellulose membranes was significantly higher than that on unmodified membranes. Cocktails of phages active against Listeria or Escherichia coli immobilized on these membranes were shown to effectively control the growth of L. monocytogenes and E. coli O157:H7 in ready-to-eat and raw meat, respectively, under different storage temperatures and packaging conditions. The phage storage stability was investigated to further extend their industrial applications. It was shown that lyophilization can be used as a phage-drying method to maintain their infectivity on the newly developed bioactive materials. In conclusion, utilizing the charge difference between phage heads and tails provided a simple technique for oriented immobilization applicable to a wide range of phages and allowed the retention of infectivity. PMID:21803890

Thylakoid membrane protein topography: transmembrane orientation of the chloroplast cytochrome b-559 psbE gene product.

PubMed

Tae, G S; Black, M T; Cramer, W A; Vallon, O; Bogorad, L

1988-12-27

Protease accessibility and antibody to a COOH-terminal peptide were used as probes for the in situ topography of the Mr 10,000 psbE gene product (alpha subunit) of the chloroplast cytochrome b-559. Exposure of thylakoid membranes to trypsin or Staphylococcus aureus V8 protease cleaved the alpha subunit to a slightly smaller polypeptide (delta Mr approximately -1000) as detected on Western blots, without loss of reactivity to COOH-terminal antibody. The disappearance of the parent Mr 10,000 polypeptide from thylakoids in the presence of trypsin correlated with the appearance of the smaller polypeptide with delta Mr = -750, the conversion having a half-time of approximately 15 min. Exposure of inside-out vesicles to trypsin resulted in almost complete loss of reactivity to the antibody, showing that the COOH terminus is exposed on the lumenal side of the membrane. Removal of the extrinsic polypeptides of the oxygen-evolving complex resulted in an increase of the accessibility of the alpha subunit to trypsin. These data establish that the alpha subunit of cytochrome b-559 crosses the membrane once, as predicted from its single, 26-residue, hydrophobic domain. The NH2 terminus of the alpha polypeptide is on the stromal side of the membrane, where it is accessible, most likely at Arg-7 or Glu-6/Asp-11, to trypsin or V8 protease, respectively. As a consequence of this orientation, the single histidine residue in the alpha subunit is located on the stromal side of the hydrophobic domain.(ABSTRACT TRUNCATED AT 250 WORDS)

DNA Molecules Adsorbed on Rippled Supported Cationic Lipid Membranes -- A new way to stretch DNAs

NASA Astrophysics Data System (ADS)

Golubovic, Leonardo

2005-03-01

We discuss a novel approach to control to shapes of DNA molecules. We elucidate the recent experimental work of M. Hochrein, L. Golubovic and J. Raedler, on the conformational behavior of DNA molecules adsorbed on lipid membranes that are supported on grooved micro-structured surfaces. We explain the striking ability of the edges formed on these supported membranes to adsorb and completely orient (stretch) very long DNA molecules. Here we explain the experimentally observed DNA stretching effect in terms of the surface curvature dependent electrostatic potential seen by the adsorbed DNA molecules. On the curved, rippled membrane, we show that the DNA molecules undergo localization transitions causing them to stretch by binding to the ripple edges of the supported membrane. In the future, this stretching will allow to directly image, by the common fluorescence microscopy, fundamental biological processes of the interactions between DNA and single protein molecules.

Orientations of Iron-Sulfur Clusters FA and FB in the Homodimeric Type-I Photosynthetic Reaction Center of Heliobacterium modesticaldum.

PubMed

Kondo, Toru; Matsuoka, Masahiro; Azai, Chihiro; Itoh, Shigeru; Oh-Oka, Hirozo

2016-05-12

Orientations of the FA and FB iron-sulfur (FeS) clusters in a structure-unknown type-I homodimeric heriobacterial reaction center (hRC) were studied in oriented membranes of the thermophilic anaerobic photosynthetic bacterium Heliobacterium modesticaldum by electron paramagnetic resonance (EPR), and compared with those in heterodimeric photosystem I (PS I). The Rieske-type FeS center in the cytochrome b/c complex showed a well-oriented EPR signal. Illumination at 14 K induced an FB(-) signal with g-axes of gz = 2.066, gy = 1.937, and gx = 1.890, tilted at angles of 60°, 60°, and 45°, respectively, with respect to the membrane normal. Chemical reduction with dithionite produced an additional signal of FA(-), which magnetically interacted with FB(-), with gz = 2.046, gy = 1.942, and gx = 1.911 at 30°, 60°, and 90°, respectively. The angles and redox properties of FA(-) and FB(-) in hRC resemble those of FB(-) and FA(-), respectively, in PS I. Therefore, FA and FB in hRC, named after their g-value similarities, seem to be located like FB and FA, not like FA and FB, respectively, in PS I. The reducing side of hRC could resemble those in PS I, if the names of FA and FB are interchanged with each other.

Agent-based modeling of malaria vectors: the importance of spatial simulation.

PubMed

Bomblies, Arne

2014-07-03

The modeling of malaria vector mosquito populations yields great insight into drivers of malaria transmission at the village scale. Simulation of individual mosquitoes as "agents" in a distributed, dynamic model domain may be greatly beneficial for simulation of spatial relationships of vectors and hosts. In this study, an agent-based model is used to simulate the life cycle and movement of individual malaria vector mosquitoes in a Niger Sahel village, with individual simulated mosquitoes interacting with their physical environment as well as humans. Various processes that are known to be epidemiologically important, such as the dependence of parity on flight distance between developmental habitat and blood meal hosts and therefore spatial relationships of pools and houses, are readily simulated using this modeling paradigm. Impacts of perturbations can be evaluated on the basis of vectorial capacity, because the interactions between individuals that make up the population- scale metric vectorial capacity can be easily tracked for simulated mosquitoes and human blood meal hosts, without the need to estimate vectorial capacity parameters. As expected, model results show pronounced impacts of pool source reduction from larvicide application and draining, but with varying degrees of impact depending on the spatial relationship between pools and human habitation. Results highlight the importance of spatially-explicit simulation that can model individuals such as in an agent-based model. The impacts of perturbations on village scale malaria transmission depend on spatial locations of individual mosquitoes, as well as the tracking of relevant life cycle events and characteristics of individual mosquitoes. This study demonstrates advantages of using an agent-based approach for village-scale mosquito simulation to address questions in which spatial relationships are known to be important.

Alternative Method for the Mass Rearing of Lutzomyia (Lutzomyia) cruzi (Diptera: Psychodidae) in a Laboratory Setting.

PubMed

Oliveira, E F; Fernandes, W S; Oshiro, E T; Oliveira, A G; Galati, E A B

2015-09-01

The understanding of the transmission dynamics of Leishmania spp. Ross as well as the epidemiology and spread of leishmaniasis is related to parasite-vector-host interactions. These interactions can be studied using specimens of a sand fly population reared in the laboratory, exposing individuals to experimental infection for the investigation of vector competence and parameters of the vectorial capacity of the species. The present study sought to describe an alternative method for the implantation of a Lutzomyia (Lutzomyia) cruzi colony with wild specimens captured in the municipality of Corumbá, Brazil. With Method 1, engorged females were individualized for oviposition. The eggs were transferred to an acrylic petri dish with a layer of plaster on the bottom, on which food was placed after hatching of the first larvae. With Method 2, females were kept in groups for oviposition in containers, in which soil and food were placed on their bottom for the larvae. In addition, the exposure time of the larvae to light was reduced in comparison with Method 1. With Method 2, a significantly greater number of specimens of Lu. cruzi was obtained. The ratio between the number of emerged adults and the females followed for oviposition was 0.42 with Method 1 and 2.75 with Method 2. The optimization of the rearing conditions for Lu. cruzi will enable the establishment of a colony providing a sufficient number of specimens to develop experimental infection by Leishmania as well as vectorial competence and some parameters of the vectorial capacity of this sand fly. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

A novel HMM distributed classifier for the detection of gait phases by means of a wearable inertial sensor network.

PubMed

Taborri, Juri; Rossi, Stefano; Palermo, Eduardo; Patanè, Fabrizio; Cappa, Paolo

2014-09-02

In this work, we decided to apply a hierarchical weighted decision, proposed and used in other research fields, for the recognition of gait phases. The developed and validated novel distributed classifier is based on hierarchical weighted decision from outputs of scalar Hidden Markov Models (HMM) applied to angular velocities of foot, shank, and thigh. The angular velocities of ten healthy subjects were acquired via three uni-axial gyroscopes embedded in inertial measurement units (IMUs) during one walking task, repeated three times, on a treadmill. After validating the novel distributed classifier and scalar and vectorial classifiers-already proposed in the literature, with a cross-validation, classifiers were compared for sensitivity, specificity, and computational load for all combinations of the three targeted anatomical segments. Moreover, the performance of the novel distributed classifier in the estimation of gait variability in terms of mean time and coefficient of variation was evaluated. The highest values of specificity and sensitivity (>0.98) for the three classifiers examined here were obtained when the angular velocity of the foot was processed. Distributed and vectorial classifiers reached acceptable values (>0.95) when the angular velocity of shank and thigh were analyzed. Distributed and scalar classifiers showed values of computational load about 100 times lower than the one obtained with the vectorial classifier. In addition, distributed classifiers showed an excellent reliability for the evaluation of mean time and a good/excellent reliability for the coefficient of variation. In conclusion, due to the better performance and the small value of computational load, the here proposed novel distributed classifier can be implemented in the real-time application of gait phases recognition, such as to evaluate gait variability in patients or to control active orthoses for the recovery of mobility of lower limb joints.

Vectorial transport of unconjugated and conjugated bile salts by monolayers of LLC-PK1 cells doubly transfected with human NTCP and BSEP or with rat Ntcp and Bsep.

PubMed

Mita, Sachiko; Suzuki, Hiroshi; Akita, Hidetaka; Hayashi, Hisamitsu; Onuki, Reiko; Hofmann, Alan F; Sugiyama, Yuichi

2006-03-01

Na(+)-taurocholate-cotransporting peptide (NTCP)/SLC10A1 and bile salt export pump (BSEP)/ABCB11 synergistically play an important role in the transport of bile salts by the hepatocyte. In this study, we transfected human NTCP and BSEP or rat Ntcp and Bsep into LLC-PK1 cells, a cell line devoid of bile salts transporters. Transport by these cells was characterized with a focus on substrate specificity between rats and humans. The basal to apical flux of taurocholate across NTCP- and BSEP-expressing LLC-PK1 monolayers was 10 times higher than that in the opposite direction, whereas the flux across the monolayer of control and NTCP or BSEP single-expressing cells did not show any vectorial transport. The basal to apical flux of taurocholate was saturated with a K(m) value of 20 microM. Vectorial transcellular transport was also observed for cholate, chenodeoxycholate, ursodeoxycholate, their taurine and glycine conjugates, and taurodeoxycholate and glycodeoxycholate, whereas no transport of lithocholate was detected. To evaluate the respective functions of NTCP and BSEP and to compare them with those of rat Ntcp and Bsep, we calculated the clearance by each transporter in this system. A good correlation in the clearance of the examined bile salts (cholate, chenodeoxycholate, ursodeoxycholate, and their taurine or glycine conjugates) was observed between transport by human and that of rat transporters in terms of their rank order: for NTCP, taurine conjugates > glycine conjugates > unconjugated bile salts, and for BSEP, unconjugated bile salts and glycine conjugates > taurine conjugates. In conclusion, the substrate specificity of human and rat NTCP and BSEP appear to be very similar at least for monovalent bile salts under physiological conditions.

Vectorial capacity of Aedes aegypti for dengue virus type 2 is reduced with co-infection of Metarhizium anisopliae.

PubMed

Garza-Hernández, Javier A; Rodríguez-Pérez, Mario A; Salazar, Ma Isabel; Russell, Tanya L; Adeleke, Monsuru A; de Luna-Santillana, Erik de J; Reyes-Villanueva, Filiberto

2013-01-01

Aedes aegypti, is the major dengue vector and a worldwide public health threat combated basically by chemical insecticides. In this study, the vectorial competence of Ae. aegypti co-infected with a mildly virulent Metarhizium anisopliae and fed with blood infected with the DENV-2 virus, was examined. The study encompassed three bioassays (B). In B1 the median lethal time (LT50) of Ae. aegypti exposed to M. anisopliae was determined in four treatments: co-infected (CI), single-fungus infection (SF), single-virus infection (SV) and control (C). In B2, the mortality and viral infection rate in midgut and in head were registered in fifty females of CI and in SV. In B3, the same treatments as in B1 but with females separated individually were tested to evaluate the effect on fecundity and gonotrophic cycle length. Survival in CI and SF females was 70% shorter than the one of those in SV and control. Overall viral infection rate in CI and SV were 76 and 84% but the mortality at day six post-infection was 78% (54% infected) and 6% respectively. Survivors with virus in head at day seven post-infection were 12 and 64% in both CI and SV mosquitoes. Fecundity and gonotrophic cycle length were reduced in 52 and 40% in CI compared to the ones in control. Fungus-induced mortality for the CI group was 78%. Of the survivors, 12% (6/50) could potentially transmit DENV-2, as opposed to 64% (32/50) of the SV group, meaning a 5-fold reduction in the number of infective mosquitoes. This is the first report on a fungus that reduces the vectorial capacity of Ae. aegypti infected with the DENV-2 virus.

An analysis of the factors influencing the residual refractive astigmatism after cataract surgery with toric intraocular lenses.

PubMed

Savini, Giacomo; Næser, Kristian

2015-01-13

To investigate the influence of posterior corneal astigmatism, surgically-induced corneal astigmatism (SICA), intraocular lens (IOL) orientation, and effective lens position on the refractive outcome of toric IOLs. Five models were prospectively investigated. Keratometric astigmatism and an intended SICA of 0.2 diopters (D) were entered into model 1. Total corneal astigmatism, measured by a rotating Scheimpflug camera, was used instead of keratometric astigmatism in model 2. The mean postoperative SICA, the actual postoperative IOL orientation, and the influence of the effective lens position were added, respectively, into models 3, 4, and 5. Astigmatic data were vectorially described by meridional and torsional powers. A set of equations was developed to describe the error in refractive astigmatism (ERA) as the difference between the postoperative refractive astigmatism and the target refractive astigmatism. We enrolled 40 consecutive eyes. In model 1, ERA calculations revealed significant cylinder overcorrection in with-the-rule (WTR) eyes (meridional power = -0.59 ± 0.34 D, P < 0.0001) and undercorrection in against-the-rule (ATR) eyes (0.32 ± 0.42 D, P = 0.01). When total corneal astigmatism was used instead of keratometric astigmatism (model 2), the ERA meridional power decreased in WTR (-0.13 ± 0.42 D) and ATR (0.07 ± 0.59 D) eyes, both values being not statistically significant. Models 3 to 5 did not lead to significant improvement. Posterior corneal astigmatism exerts the highest influence on the ERA after toric IOL implantation. Basing calculations on total corneal astigmatism rather than keratometric astigmatism improves the prediction of the residual refractive astigmatism. Copyright 2015 The Association for Research in Vision and Ophthalmology, Inc.

Membrane tension and cytoskeleton organization in cell motility.

PubMed

Sens, Pierre; Plastino, Julie

2015-07-15

Cell membrane shape changes are important for many aspects of normal biological function, such as tissue development, wound healing and cell division and motility. Various disease states are associated with deregulation of how cells move and change shape, including notably tumor initiation and cancer cell metastasis. Cell motility is powered, in large part, by the controlled assembly and disassembly of the actin cytoskeleton. Much of this dynamic happens in close proximity to the plasma membrane due to the fact that actin assembly factors are membrane-bound, and thus actin filaments are generally oriented such that their growth occurs against or near the membrane. For a long time, the membrane was viewed as a relatively passive scaffold for signaling. However, results from the last five years show that this is not the whole picture, and that the dynamics of the actin cytoskeleton are intimately linked to the mechanics of the cell membrane. In this review, we summarize recent findings concerning the role of plasma membrane mechanics in cell cytoskeleton dynamics and architecture, showing that the cell membrane is not just an envelope or a barrier for actin assembly, but is a master regulator controlling cytoskeleton dynamics and cell polarity.

Computing Curvature Sensitivity of Biomolecules in Membranes by Simulated Buckling.

PubMed

Elías-Wolff, Federico; Lindén, Martin; Lyubartsev, Alexander P; Brandt, Erik G

2018-03-13

Membrane curvature sensing, where the binding free energies of membrane-associated molecules depend on the local membrane curvature, is a key factor to modulate and maintain the shape and organization of cell membranes. However, the microscopic mechanisms are not well understood, partly due to absence of efficient simulation methods. Here, we describe a method to compute the curvature dependence of the binding free energy of a membrane-associated probe molecule that interacts with a buckled membrane, which has been created by lateral compression of a flat bilayer patch. This buckling approach samples a wide range of curvatures in a single simulation, and anisotropic effects can be extracted from the orientation statistics. We develop an efficient and robust algorithm to extract the motion of the probe along the buckled membrane surface, and evaluate its numerical properties by extensive sampling of three coarse-grained model systems: local lipid density in a curved environment for single-component bilayers, curvature preferences of individual lipids in two-component membranes, and curvature sensing by a homotrimeric transmembrane protein. The method can be used to complement experimental data from curvature partition assays and provides additional insight into mesoscopic theories and molecular mechanisms for curvature sensing.

Effects of stretching and stirring on water and glucose absorption by canine mucosal membrane.

PubMed Central

Lee, J S

1983-01-01

A 'mini' canine mucosal membrane preparation permitting simultaneous determination of water (Jv) and glucose (Jg) absorption rates, microscopic examination or micropuncture of the villi was used in this study. The small membranes were more stretched than the large ones, with more than a one-fold increase in both Jv and Jg, apparently due to a change in architectural orientation between the villi and subvillous supporting tissue so as to facilitate water transport via the lymphatic system. During stirring of the bathing solution, the villi in the small membranes were widely separated from each other with more to-and-fro swaying movements than in the large ones. Stirring was seen to cause up-and-down movements of the loosely suspended large membranes but not the small ones. In the small membranes stirring caused no change in Jv but an increase in Jg due to the increase in glucose concentration in the absorbate, while in the large membranes both Jv and Jg were greatly increased. It is thus considered that the increase in absorption in the large membranes caused by stirring is mainly due to the increased membrane movements promoting lymph flow. PMID:6875881

Physiologically-Relevant Modes of Membrane Interactions by the Human Antimicrobial Peptide, LL-37, Revealed by SFG Experiments

PubMed Central

Ding, Bei; Soblosky, Lauren; Nguyen, Khoi; Geng, Junqing; Yu, Xinglong; Ramamoorthy, Ayyalusamy; Chen, Zhan

2013-01-01

Antimicrobial peptides (AMPs) could become the next generation antibiotic compounds which can overcome bacterial resistance by disrupting cell membranes and it is essential to determine the factors underlying its mechanism of action. Although high-resolution NMR and other biological studies have provided valuable insights, it has been a major challenge to follow the AMP-membrane interactions at physiologically-relevant low peptide concentrations. In this study, we demonstrate a novel approach to overcome this major limitation by performing Sum Frequency Generation (SFG) vibrational spectroscopic experiments on lipid bilayers containing an AMP, LL-37. Our results demonstrate the power of SFG to study non-linear helical peptides and also infer that lipid-peptide interaction and the peptide orientation depend on the lipid membrane composition. The observed SFG signal changes capture the aggregating process of LL-37 on membrane. In addition, our SFG results on cholesterol-containing lipid bilayers indicate the inhibition effect of cholesterol on peptide-induced membrane permeation process. PMID:23676762

Physiologically-Relevant Modes of Membrane Interactions by the Human Antimicrobial Peptide, LL-37, Revealed by SFG Experiments

NASA Astrophysics Data System (ADS)

Ding, Bei; Soblosky, Lauren; Nguyen, Khoi; Geng, Junqing; Yu, Xinglong; Ramamoorthy, Ayyalusamy; Chen, Zhan

2013-05-01

Antimicrobial peptides (AMPs) could become the next generation antibiotic compounds which can overcome bacterial resistance by disrupting cell membranes and it is essential to determine the factors underlying its mechanism of action. Although high-resolution NMR and other biological studies have provided valuable insights, it has been a major challenge to follow the AMP-membrane interactions at physiologically-relevant low peptide concentrations. In this study, we demonstrate a novel approach to overcome this major limitation by performing Sum Frequency Generation (SFG) vibrational spectroscopic experiments on lipid bilayers containing an AMP, LL-37. Our results demonstrate the power of SFG to study non-linear helical peptides and also infer that lipid-peptide interaction and the peptide orientation depend on the lipid membrane composition. The observed SFG signal changes capture the aggregating process of LL-37 on membrane. In addition, our SFG results on cholesterol-containing lipid bilayers indicate the inhibition effect of cholesterol on peptide-induced membrane permeation process.

Physiologically-relevant modes of membrane interactions by the human antimicrobial peptide, LL-37, revealed by SFG experiments.

PubMed

Ding, Bei; Soblosky, Lauren; Nguyen, Khoi; Geng, Junqing; Yu, Xinglong; Ramamoorthy, Ayyalusamy; Chen, Zhan

2013-01-01

Antimicrobial peptides (AMPs) could become the next generation antibiotic compounds which can overcome bacterial resistance by disrupting cell membranes and it is essential to determine the factors underlying its mechanism of action. Although high-resolution NMR and other biological studies have provided valuable insights, it has been a major challenge to follow the AMP-membrane interactions at physiologically-relevant low peptide concentrations. In this study, we demonstrate a novel approach to overcome this major limitation by performing Sum Frequency Generation (SFG) vibrational spectroscopic experiments on lipid bilayers containing an AMP, LL-37. Our results demonstrate the power of SFG to study non-linear helical peptides and also infer that lipid-peptide interaction and the peptide orientation depend on the lipid membrane composition. The observed SFG signal changes capture the aggregating process of LL-37 on membrane. In addition, our SFG results on cholesterol-containing lipid bilayers indicate the inhibition effect of cholesterol on peptide-induced membrane permeation process.

Entropy and biological systems: experimentally-investigated entropy-driven stacking of plant photosynthetic membranes.

PubMed

Jia, Husen; Liggins, John R; Chow, Wah Soon

2014-02-24

According to the Second Law of Thermodynamics, an overall increase of entropy contributes to the driving force for any physicochemical process, but entropy has seldom been investigated in biological systems. Here, for the first time, we apply Isothermal Titration Calorimetry (ITC) to investigate the Mg(2+)-induced spontaneous stacking of photosynthetic membranes isolated from spinach leaves. After subtracting a large endothermic interaction of MgCl₂ with membranes, unrelated to stacking, we demonstrate that the enthalpy change (heat change at constant pressure) is zero or marginally positive or negative. This first direct experimental evidence strongly suggests that an entropy increase significantly drives membrane stacking in this ordered biological structure. Possible mechanisms for the entropy increase include: (i) the attraction between discrete oppositely-charged areas, releasing counterions; (ii) the release of loosely-bound water molecules from the inter-membrane gap; (iii) the increased orientational freedom of previously-aligned water dipoles; and (iv) the lateral rearrangement of membrane components.

Purification and proteomic analysis of plant plasma membranes.

PubMed

Alexandersson, Erik; Gustavsson, Niklas; Bernfur, Katja; Karlsson, Adine; Kjellbom, Per; Larsson, Christer

2008-01-01

All techniques needed for proteomic analyses of plant plasma membranes are described in detail, from isolation of plasma membranes to protein identification by mass spectrometry (MS). Plasma membranes are isolated by aqueous two-phase partitioning yielding vesicles with a cytoplasmic side-in orientation and a purity of about 95%. These vesicles are turned inside-out by treatment with Brij 58, which removes soluble contaminating proteins enclosed in the vesicles as well as loosely attached proteins. The final plasma membrane preparation thus retains all integral proteins and many peripheral proteins. Proteins are separated by one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), and protein bands are excised and digested with trypsin. Peptides in tryptic digests are separated by nanoflow liquid chromatography and either fed directly into an ESI-MS or spotted onto matrix-assisted laser desorption ionization (MALDI) plates for analysis with MALDI-MS. Finally, data processing and database searching are used for protein identification to define a plasma membrane proteome.

Entropy and biological systems: Experimentally-investigated entropy-driven stacking of plant photosynthetic membranes

PubMed Central

Jia, Husen; Liggins, John R.; Chow, Wah Soon

2014-01-01

According to the Second Law of Thermodynamics, an overall increase of entropy contributes to the driving force for any physicochemical process, but entropy has seldom been investigated in biological systems. Here, for the first time, we apply Isothermal Titration Calorimetry (ITC) to investigate the Mg2+-induced spontaneous stacking of photosynthetic membranes isolated from spinach leaves. After subtracting a large endothermic interaction of MgCl2 with membranes, unrelated to stacking, we demonstrate that the enthalpy change (heat change at constant pressure) is zero or marginally positive or negative. This first direct experimental evidence strongly suggests that an entropy increase significantly drives membrane stacking in this ordered biological structure. Possible mechanisms for the entropy increase include: (i) the attraction between discrete oppositely-charged areas, releasing counterions; (ii) the release of loosely-bound water molecules from the inter-membrane gap; (iii) the increased orientational freedom of previously-aligned water dipoles; and (iv) the lateral rearrangement of membrane components. PMID:24561561

Self-assembling layers created by membrane proteins on gold.

PubMed

Shah, D S; Thomas, M B; Phillips, S; Cisneros, D A; Le Brun, A P; Holt, S A; Lakey, J H

2007-06-01

Membrane systems are based on several types of organization. First, amphiphilic lipids are able to create monolayer and bilayer structures which may be flat, vesicular or micellar. Into these structures membrane proteins can be inserted which use the membrane to provide signals for lateral and orientational organization. Furthermore, the proteins are the product of highly specific self-assembly otherwise known as folding, which mostly places individual atoms at precise places in three dimensions. These structures all have dimensions in the nanoscale, except for the size of membrane planes which may extend for millimetres in large liposomes or centimetres on planar surfaces such as monolayers at the air/water interface. Membrane systems can be assembled on to surfaces to create supported bilayers and these have uses in biosensors and in electrical measurements using modified ion channels. The supported systems also allow for measurements using spectroscopy, surface plasmon resonance and atomic force microscopy. By combining the roles of lipids and proteins, highly ordered and specific structures can be self-assembled in aqueous solution at the nanoscale.

A Neutron View of Proteins in Lipid Bilayers

NASA Astrophysics Data System (ADS)

White, Stephen

2012-02-01

Despite the growing number of atomic-resolution membrane protein structures, direct structural information about proteins in their native membrane environment is scarce. This problem is particularly relevant in the case of the highly-charged S1-S4 voltage- sensing domains responsible for nerve impulses, where interactions with the lipid bilayer are critical for the function of voltage-activated potassium channels. We have used neutron diffraction, solid-state nuclear magnetic resonance spectroscopy, and molecular dynamics simulations to investigate the structure and hydration of bilayer membranes containing S1-S4 voltage-sensing domains. Our results show that voltage sensors adopt transmembrane orientations, cause a modest reshaping of the surrounding lipid bilayer, and that water molecules intimately interact with the protein within the membrane. These structural findings reveal that voltage sensors have evolved to interact with the lipid membrane while keeping the energetic and structural perturbations to a minimum, and that water penetrates into the membrane to hydrate charged residues and shape the transmembrane electric field.

Enhanced antibacterial activity through the controlled alignment of graphene oxide nanosheets.

PubMed

Lu, Xinglin; Feng, Xunda; Werber, Jay R; Chu, Chiheng; Zucker, Ines; Kim, Jae-Hong; Osuji, Chinedum O; Elimelech, Menachem

2017-11-14

The cytotoxicity of 2D graphene-based nanomaterials (GBNs) is highly important for engineered applications and environmental health. However, the isotropic orientation of GBNs, most notably graphene oxide (GO), in previous experimental studies obscured the interpretation of cytotoxic contributions of nanosheet edges. Here, we investigate the orientation-dependent interaction of GBNs with bacteria using GO composite films. To produce the films, GO nanosheets are aligned in a magnetic field, immobilized by cross-linking of the surrounding matrix, and exposed on the surface through oxidative etching. Characterization by small-angle X-ray scattering and atomic force microscopy confirms that GO nanosheets align progressively well with increasing magnetic field strength and that the alignment is effectively preserved by cross-linking. When contacted with the model bacterium Escherichia coli , GO nanosheets with vertical orientation exhibit enhanced antibacterial activity compared with random and horizontal orientations. Further characterization is performed to explain the enhanced antibacterial activity of the film with vertically aligned GO. Using phospholipid vesicles as a model system, we observe that GO nanosheets induce physical disruption of the lipid bilayer. Additionally, we find substantial GO-induced oxidation of glutathione, a model intracellular antioxidant, paired with limited generation of reactive oxygen species, suggesting that oxidation occurs through a direct electron-transfer mechanism. These physical and chemical mechanisms both require nanosheet penetration of the cell membrane, suggesting that the enhanced antibacterial activity of the film with vertically aligned GO stems from an increased density of edges with a preferential orientation for membrane disruption. The importance of nanosheet penetration for cytotoxicity has direct implications for the design of engineering surfaces using GBNs.

Large-scale fabrication of single crystalline tin nanowire arrays.

PubMed

Luo, Bin; Yang, Dachi; Liang, Minghui; Zhi, Linjie

2010-09-01

Large-scale single crystalline tin nanowire arrays with preferred lattice orientation along the [100] direction were fabricated in porous anodic aluminium oxide (AAO) membranes by the electrodeposition method using copper nanorod as a second electrode.

Principles and biophysical applications of single particle super-localization and rotational tracking

NASA Astrophysics Data System (ADS)

Gu, Yan

While conventional Single Particle Tracking (SPT) techniques acquire 2D or 3D trajectories of particle probes, we have developed Single Particle Orientation and Rotational Tracking (SPORT) techniques to extract orientation and rotational information. Combined with DIC microscopy, the SPORT technique has been applied in biophysical studies, including membrane diffusion and intracellular transport. The rotational dynamics of nanoparticle vectors on live cell membranes was recorded and its influence on the fate of these nanoparticle vectors was elucidated. The rotational motions of gold nanorods with various surface modifiers were tracked continuously at a temporal resolution of 5 ms under a DIC microscope. We found that the rotational behaviors of gold nanorod vectors are strongly related to their surface charge, specific surface functional groups, and the availability of receptors on cell membranes. The study of rotational Brownian motion of nanoparticles on cell membranes will lead to a better understanding of the mechanisms of drug delivery and provide guidance in designing surface modification strategies for drug delivery vectors under various circumstances. To characterize the rotation mode of surface functionalized gold nanorods on cell membranes, the SPORT technique is combined with the correlation analysis of the bright and dark DIC intensities. The unique capabilities of visualizing and understanding rotational motions of functionalized nanoparticles on live cell membranes allow us to correlate rotational and translational dynamics in unprecedented detail and provide new insights for complex membrane processes, including electrostatic interactions, ligand-receptor binding, and lateral (confined and hopping) diffusion of membrane receptors. Surface-functionalized nanoparticles interact with the membrane in fundamentally different ways and exhibit distinct rotational modes. The early events of particle-membrane approach and attachment are directly visualized for the first time. The rotational dynamics of cargos in both active directional transport and pausing stages of axonal transport was also visualized using high-speed SPORT with a temporal resolution of 2 ms. Both long and short pauses are imaged, and the correlations between the pause duration, the rotational behaviour of the cargo at the pause, and the moving direction after the pause are established. Furthermore, the rotational dynamics leading to switching tracks are visualized in detail. These first-time observations of cargo's rotational dynamics provide new insights on how kinesin and dynein motors take the cargo through the alternating stages of active directional transport and pause. To improve the localization precision of the SPT technique with DIC microscopy, a precise three-dimensional (3D) localization method of spherical gold nanoparticle probes using model-based correlation coefficient mapping was introduced. To accomplish this, a stack of sample images at different z-positions are acquired, and a 3D intensity profile of the probe serving as the model is used to map out the positions of nanoparticles in the sample. By using this model-based correlation imaging method, precise localization can be achieved in imaging techniques with complicated point spread functions (PSF) such as differential interference contrast (DIC) microscopy. The 3D superlocalization method was applied to tracking gold nanospheres during live endocytosis events. Finally, a novel dual-modality imaging technique has been developed to super-localize a single gold nanorod while providing its orientation and rotational information. The super-localization of the gold nanorod can be accomplished by curve fitting the modified bright-field images generated by one of the two beams laterally shifted by the first Nomarski prism in a DIC microscope. The orientation and rotational information is derived from the DIC images of gold nanorods. The new imaging setup has been applied to study the steric hindrance induced by relatively large cargos in the microtubule gliding assay and to track nanocargos in the crowded cellular environment.

Principles and biophysical applications of single particle super-localization and rotational tracking

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

Gu, Yan

While conventional Single Particle Tracking (SPT) techniques acquire 2D or 3D trajectories of particle probes, we have developed Single Particle Orientation and Rotational Tracking (SPORT) techniques to extract orientation and rotational information. Combined with DIC microscopy, the SPORT technique has been applied in biophysical studies, including membrane diffusion and intracellular transport. The rotational dynamics of nanoparticle vectors on live cell membranes was recorded and its influence on the fate of these nanoparticle vectors was elucidated. The rotational motions of gold nanorods with various surface modifiers were tracked continuously at a temporal resolution of 5 ms under a DIC microscope. Wemore » found that the rotational behaviors of gold nanorod vectors are strongly related to their surface charge, specific surface functional groups, and the availability of receptors on cell membranes. The study of rotational Brownian motion of nanoparticles on cell membranes will lead to a better understanding of the mechanisms of drug delivery and provide guidance in designing surface modification strategies for drug delivery vectors under various circumstances. To characterize the rotation mode of surface functionalized gold nanorods on cell membranes, the SPORT technique is combined with the correlation analysis of the bright and dark DIC intensities. The unique capabilities of visualizing and understanding rotational motions of functionalized nanoparticles on live cell membranes allow us to correlate rotational and translational dynamics in unprecedented detail and provide new insights for complex membrane processes, including electrostatic interactions, ligand-receptor binding, and lateral (confined and hopping) diffusion of membrane receptors. Surface-functionalized nanoparticles interact with the membrane in fundamentally different ways and exhibit distinct rotational modes. The early events of particle-membrane approach and attachment are directly visualized for the first time. The rotational dynamics of cargos in both active directional transport and pausing stages of axonal transport was also visualized using high-speed SPORT with a temporal resolution of 2 ms. Both long and short pauses are imaged, and the correlations between the pause duration, the rotational behaviour of the cargo at the pause, and the moving direction after the pause are established. Furthermore, the rotational dynamics leading to switching tracks are visualized in detail. These first-time observations of cargo's rotational dynamics provide new insights on how kinesin and dynein motors take the cargo through the alternating stages of active directional transport and pause. To improve the localization precision of the SPT technique with DIC microscopy, a precise three-dimensional (3D) localization method of spherical gold nanoparticle probes using model-based correlation coefficient mapping was introduced. To accomplish this, a stack of sample images at different z-positions are acquired, and a 3D intensity profile of the probe serving as the model is used to map out the positions of nanoparticles in the sample. By using this model-based correlation imaging method, precise localization can be achieved in imaging techniques with complicated point spread functions (PSF) such as differential interference contrast (DIC) microscopy. The 3D superlocalization method was applied to tracking gold nanospheres during live endocytosis events. Finally, a novel dual-modality imaging technique has been developed to super-localize a single gold nanorod while providing its orientation and rotational information. The super-localization of the gold nanorod can be accomplished by curve fitting the modified bright-field images generated by one of the two beams laterally shifted by the first Nomarski prism in a DIC microscope. The orientation and rotational information is derived from the DIC images of gold nanorods. The new imaging setup has been applied to study the steric hindrance induced by relatively large cargos in the microtubule gliding assay and to track nanocargos in the crowded cellular environment.« less

The role of a Sertoli cell actin-myosin system in sperm bundle formation in the ratfish, Hydrolagus colliei (Chondrichthyes, Holocephali).

PubMed

Stanley, H P; Lambert, C C

1985-11-01

Sertoli cells in the ratfish entirely surround a clone of spermatids to form a spermatocyst. As spermiogenesis proceeds within the cyst cavity, the acrosome areas become apposed to the Sertoli cell plasma membrane lining the spermatocyst. The spermatids elongate and are gathered into an increasingly compact bundle oriented with acrosomal tips directed toward the Sertoli cell base. As all acrosome areas move closer together, Sertoli cell microfilaments oriented parallel to the long spermatid axis appear and increase in concentration. Actin and myosin were demonstrated in the microfilament area with fluorescent antibodies and NBD-Phallacidin. Simultaneously, endocytosis of Sertoli cell membrane between spermatid attachment sites removes the intervening membrane and allows the latter sites to approach each other. Sertoli cell endocytosis is spatially and temporally related to a unique projection at the basal rim of each acrosome. During midspermiogenesis, structured intercellular material appears between the Sertoli cell and the acrosomal region of each spermatid. Its periodicity is closely related to periodic arrangement of Sertoli cell actin and material within the spermatids. These attachment sites move together upon endocytosis, gathering a clone of spermatids into a closely packed bundle.

Reconstitution of SNARE proteins into solid-supported lipid bilayer stacks and X-ray structure analysis.

PubMed

Xu, Yihui; Kuhlmann, Jan; Brennich, Martha; Komorowski, Karlo; Jahn, Reinhard; Steinem, Claudia; Salditt, Tim

2018-02-01

SNAREs are known as an important family of proteins mediating vesicle fusion. For various biophysical studies, they have been reconstituted into supported single bilayers via proteoliposome adsorption and rupture. In this study we extended this method to the reconstitution of SNAREs into supported multilamellar lipid membranes, i.e. oriented multibilayer stacks, as an ideal model system for X-ray structure analysis (X-ray reflectivity and diffraction). The reconstitution was implemented through a pathway of proteomicelle, proteoliposome and multibilayer. To monitor the structural evolution in each step, we used small-angle X-ray scattering for the proteomicelles and proteoliposomes, followed by X-ray reflectivity and grazing-incidence small-angle scattering for the multibilayers. Results show that SNAREs can be successfully reconstituted into supported multibilayers, with high enough orientational alignment for the application of surface sensitive X-ray characterizations. Based on this protocol, we then investigated the effect of SNAREs on the structure and phase diagram of the lipid membranes. Beyond this application, this reconstitution protocol could also be useful for X-ray analysis of many further membrane proteins. Copyright © 2017 Elsevier B.V. All rights reserved.

Bioecology and vectorial capacity of Aedes albopictus (Diptera: Culicidae) in Macao, China, in relation to dengue virus transmission.

PubMed

Almeida, A Paulo G; Baptista, Susana S S G; Sousa, Carla A G C C; Novo, M Teresa L M; Ramos, Helena C; Panella, Nicholas A; Godsey, Marvin; Simões, M João; Anselmo, M Luisa; Komar, Nicholas; Mitchell, Carl J; Ribeiro, Henrique

2005-05-01

Until 2001, the Chinese Territory of Macao had not registered any autochthonous dengue cases, despite the abundance of Aedes albopictus (Skuse), a known vector. This work describes a bioecological characterization of the local Ae. albopictus adult population, with the purpose of estimating the receptivity of Macao to dengue introduction. In the wet seasons of 1997 and 1998 and the dry season of 1998, Ae. albopictus was the most abundant human-biting mosquito. Daily biting rates of 314 mosquitoes per person were recorded in the wet season with a reduction to 94 in the dry season. Ae. albopictus was mainly exophagic and exophilic and had a human blood index of 44%. The parity rate of field-collected mosquitoes was 57%. Daily survival rate ranged from 91 to 97%. Estimates of vectorial capacity ranged from 144 to 880, depending on what parameter values were used. These estimates indicated a great receptivity for the introduction of dengue viruses, as the 2001 outbreak came to prove.

A vectorial semantics approach to personality assessment.

PubMed

Neuman, Yair; Cohen, Yochai

2014-04-23

Personality assessment and, specifically, the assessment of personality disorders have traditionally been indifferent to computational models. Computational personality is a new field that involves the automatic classification of individuals' personality traits that can be compared against gold-standard labels. In this context, we introduce a new vectorial semantics approach to personality assessment, which involves the construction of vectors representing personality dimensions and disorders, and the automatic measurements of the similarity between these vectors and texts written by human subjects. We evaluated our approach by using a corpus of 2468 essays written by students who were also assessed through the five-factor personality model. To validate our approach, we measured the similarity between the essays and the personality vectors to produce personality disorder scores. These scores and their correspondence with the subjects' classification of the five personality factors reproduce patterns well-documented in the psychological literature. In addition, we show that, based on the personality vectors, we can predict each of the five personality factors with high accuracy.

Rigorous theory of the diffraction of Gaussian beams by finite gratings: TM polarization.

PubMed

Mata-Mendez, O; Avendaño, J; Chavez-Rivas, F

2006-08-01

Diffraction of TM-polarized Gaussian beams by N equally spaced slits (finite grating) in a planar perfectly conducting thick screen is treated. We extend to the TM polarization case the results of a previous paper where the TE polarization was considered. The far-field diffraction patterns, the transmission coefficient tau, and the normally diffracted energy E as a function of several optogeometrical parameters are analyzed within the so-called vectorial region. The existence of constant-intensity angles in the far field when the incident beam wave is scanned along the N slits is shown. In addition, the property E=Ntau/lambda, valid in the scalar region, is extended to the TM polarization case in the vectorial region, lambda being the wavelength. The coupling between slits is analyzed, giving an oscillating amplitude-decreasing function as the separation between slits increases, where the period for these oscillations is the wavelength lambda. Finally, the extraordinary optical transmission phenomena that appear when the wavelength is larger than the slit width (subwavelength regime) are analyzed.

Vectorial method used to monitor an evolving system: Titanium oxide thin films under UV illumination

NASA Astrophysics Data System (ADS)

Béchu, Solène; Humbert, Bernard; Fernandez, Vincent; Fairley, Neal; Richard-Plouet, Mireille

2018-07-01

Under in situ UV illumination, some materials present evolution of their opto-electronic properties that can be monitored by spectroscopy. We present here a mathematical method which can be applied to spectroscopic measurements when an evolving set of data is recorded: the vectorial method. The investigations and quantifications are performed by Infrared spectroscopy and XPS on organic-inorganic thin films prepared by sol-gel. The inorganic part of these hybrid thin films contains Ti oxide-network based whereas the organic part is composed of N,N-dimethylformamide and its hydrolysis products. Under UV illumination, those films exhibit intermediate bandgap behavior due to the photoreduction of Ti(IV) in Ti(III). The role of the solvent in the thin film is underlined during the process of photoreduction together with an understanding of the condensation of the Ti oxide-based network, as these evolutions are critical for the opto-electronic properties of those thin films.

A Vectorial Semantics Approach to Personality Assessment

NASA Astrophysics Data System (ADS)

Neuman, Yair; Cohen, Yochai

2014-04-01

Personality assessment and, specifically, the assessment of personality disorders have traditionally been indifferent to computational models. Computational personality is a new field that involves the automatic classification of individuals' personality traits that can be compared against gold-standard labels. In this context, we introduce a new vectorial semantics approach to personality assessment, which involves the construction of vectors representing personality dimensions and disorders, and the automatic measurements of the similarity between these vectors and texts written by human subjects. We evaluated our approach by using a corpus of 2468 essays written by students who were also assessed through the five-factor personality model. To validate our approach, we measured the similarity between the essays and the personality vectors to produce personality disorder scores. These scores and their correspondence with the subjects' classification of the five personality factors reproduce patterns well-documented in the psychological literature. In addition, we show that, based on the personality vectors, we can predict each of the five personality factors with high accuracy.

Three-dimensional polarization marked multiple-QR code encryption by optimizing a single vectorial beam

NASA Astrophysics Data System (ADS)

Lin, Chao; Shen, Xueju; Hua, Binbin; Wang, Zhisong

2015-10-01

We demonstrate the feasibility of three dimensional (3D) polarization multiplexing by optimizing a single vectorial beam using a multiple-signal window multiple-plane (MSW-MP) phase retrieval algorithm. Original messages represented with multiple quick response (QR) codes are first partitioned into a series of subblocks. Then, each subblock is marked with a specific polarization state and randomly distributed in 3D space with both longitudinal and transversal adjustable freedoms. A generalized 3D polarization mapping protocol is established to generate a 3D polarization key. Finally, multiple-QR code is encrypted into one phase only mask and one polarization only mask based on the modified Gerchberg-Saxton (GS) algorithm. We take the polarization mask as the cyphertext and the phase only mask as additional dimension of key. Only when both the phase key and 3D polarization key are correct, original messages can be recovered. We verify our proposal with both simulation and experiment evidences.

LOPES-3D: An antenna array for full signal detection of air-shower radio emission

NASA Astrophysics Data System (ADS)

Apel, W. D.; Arteaga, J. C.; Bähren, L.; Bekk, K.; Bertaina, M.; Biermann, P. L.; Blümer, J.; Bozdog, H.; Brancus, I. M.; Buchholz, P.; Cantoni, E.; Chiavassa, A.; Daumiller, K.; de Souza, V.; Di Pierro, F.; Doll, P.; Engel, R.; Falcke, H.; Finger, M.; Fuchs, B.; Fuhrmann, D.; Gemmeke, H.; Grupen, C.; Haungs, A.; Heck, D.; Hörandel, J. R.; Horneffer, A.; Huber, D.; Huege, T.; Isar, P. G.; Kampert, K.-H.; Kang, D.; Krömer, O.; Kuijpers, J.; Link, K.; Łuczak, P.; Ludwig, M.; Mathes, H. J.; Melissas, M.; Morello, C.; Oehlschläger, J.; Palmieri, N.; Pierog, T.; Rautenberg, J.; Rebel, H.; Roth, M.; Rühle, C.; Saftoiu, A.; Schieler, H.; Schmidt, A.; Schröder, F. G.; Sima, O.; Toma, G.; Trinchero, G. C.; Weindl, A.; Wochele, J.; Wommer, M.; Zabierowski, J.; Zensus, J. A.

2012-12-01

To better understand the radio signal emitted by extensive air-showers and to further develop the radio detection technique of high-energy cosmic rays, the LOPES experiment was reconfigured to LOPES-3D. LOPES-3D is able to measure all three vectorial components of the electric field of radio emission from cosmic ray air showers. The additional measurement of the vertical component ought to increase the reconstruction accuracy of primary cosmic ray parameters like direction and energy, provides an improved sensitivity to inclined showers, and will help to validate simulation of the emission mechanisms in the atmosphere. LOPES-3D will evaluate the feasibility of vectorial measurements for large scale applications. In order to measure all three electric field components directly, a tailor-made antenna type (tripoles) was deployed. The change of the antenna type necessitated new pre-amplifiers and an overall recalibration. The reconfiguration and the recalibration procedure are presented and the operationality of LOPES-3D is demonstrated.

A Vectorial Semantics Approach to Personality Assessment

PubMed Central

Neuman, Yair; Cohen, Yochai

2014-01-01

Personality assessment and, specifically, the assessment of personality disorders have traditionally been indifferent to computational models. Computational personality is a new field that involves the automatic classification of individuals' personality traits that can be compared against gold-standard labels. In this context, we introduce a new vectorial semantics approach to personality assessment, which involves the construction of vectors representing personality dimensions and disorders, and the automatic measurements of the similarity between these vectors and texts written by human subjects. We evaluated our approach by using a corpus of 2468 essays written by students who were also assessed through the five-factor personality model. To validate our approach, we measured the similarity between the essays and the personality vectors to produce personality disorder scores. These scores and their correspondence with the subjects' classification of the five personality factors reproduce patterns well-documented in the psychological literature. In addition, we show that, based on the personality vectors, we can predict each of the five personality factors with high accuracy. PMID:24755833

Early warnings of the potential for malaria transmission in Rural Africa using the Hydrology, Entomology and Malaria Transmission Simulator (HYDREMATS)

NASA Astrophysics Data System (ADS)

Yamana, T. K.; Eltahir, E. A.

2010-12-01

Early warnings of malaria transmission allow health officials to better prepare for future epidemics. Monitoring rainfall is recognized as an important part of malaria early warning systems, as outlined by the Roll Back Malaria Initiative. The Hydrology, Entomology and Malaria Simulator (HYDREMATS) is a mechanistic model that relates rainfall to malaria transmission, and could be used to provide early warnings of malaria epidemics. HYDREMATS is used to make predictions of mosquito populations and vectorial capacity for 2005, 2006, and 2007 in Banizoumbou village in western Niger. HYDREMATS is forced by observed rainfall, followed by a rainfall prediction based on the seasonal mean rainfall for a period two or four weeks into the future. Predictions made using this method provided reasonable estimates of mosquito populations and vectorial capacity, two to four weeks in advance. The predictions were significantly improved compared to those made when HYDREMATS was forced with seasonal mean rainfall alone.

Athermally photoreduced graphene oxides for three-dimensional holographic images

PubMed Central

Li, Xiangping; Ren, Haoran; Chen, Xi; Liu, Juan; Li, Qin; Li, Chengmingyue; Xue, Gaolei; Jia, Jia; Cao, Liangcai; Sahu, Amit; Hu, Bin; Wang, Yongtian; Jin, Guofan; Gu, Min

2015-01-01

The emerging graphene-based material, an atomic layer of aromatic carbon atoms with exceptional electronic and optical properties, has offered unprecedented prospects for developing flat two-dimensional displaying systems. Here, we show that reduced graphene oxide enabled write-once holograms for wide-angle and full-colour three-dimensional images. This is achieved through the discovery of subwavelength-scale multilevel optical index modulation of athermally reduced graphene oxides by a single femtosecond pulsed beam. This new feature allows for static three-dimensional holographic images with a wide viewing angle up to 52 degrees. In addition, the spectrally flat optical index modulation in reduced graphene oxides enables wavelength-multiplexed holograms for full-colour images. The large and polarization-insensitive phase modulation over π in reduced graphene oxide composites enables to restore vectorial wavefronts of polarization discernible images through the vectorial diffraction of a reconstruction beam. Therefore, our technique can be leveraged to achieve compact and versatile holographic components for controlling light. PMID:25901676

Biology, diversity and strategies for the monitoring and control of triatomines--Chagas disease vectors.

PubMed

Costa, Jane; Lorenzo, Marcelo

2009-07-01

Despite the relevant achievements in the control of the main Chagas disease vectors Triatoma infestans and Rhodnius prolixus, several factors still promote the risk of infection. The disease is a real threat to the poor rural regions of several countries in Latin America. The current situation in Brazil requires renewed attention due to its high diversity of triatomine species and to the rapid and drastic environmental changes that are occurring. Using the biology, behaviour and diversity of triatomines as a basis for new strategies for monitoring and controlling the vectorial transmission are discussed here. The importance of ongoing long-term monitoring activities for house infestations by T. infestans, Triatoma brasiliensis, Panstrongylus megistus, Triatoma rubrovaria and R. prolixus is also stressed, as well as understanding the invasion by sylvatic species. Moreover, the insecticide resistance is analysed. Strong efforts to sustain and improve surveillance procedures are crucial, especially when the vectorial transmission is considered interrupted in many endemic areas.

Invasiveness of Aedes aegypti and Aedes albopictus and Vectorial Capacity for Chikungunya Virus

PubMed Central

Lounibos, Leon Philip; Kramer, Laura D.

2016-01-01

In this review, we highlight biological characteristics of Aedes aegypti and Aedes albopictus, 2 invasive mosquito species and primary vectors of chikungunya virus (CHIKV), that set the tone of these species' invasiveness, vector competence, and vectorial capacity (VC). The invasiveness of both species, as well as their public health threats as vectors, is enhanced by preference for human blood. Vector competence, characterized by the efficiency of an ingested arbovirus to replicate and become infectious in the mosquito, depends largely on vector and virus genetics, and most A. aegypti and A. albopictus populations thus far tested confer vector competence for CHIKV. VC, an entomological analog of the pathogen's basic reproductive rate (R0), is epidemiologically more important than vector competence but less frequently measured, owing to challenges in obtaining valid estimates of parameters such as vector survivorship and host feeding rates. Understanding the complexities of these factors will be pivotal in curbing CHIKV transmission. PMID:27920173

Frizzled Receptors in Development and Disease

PubMed Central

Wang, Yanshu; Chang, Hao; Rattner, Amir; Nathans, Jeremy

2016-01-01

Frizzled proteins are the principal receptors for the Wnt family of ligands. They mediate canonical Wnt signaling together with Lrp5 and Lrp6 coreceptors. In conjunction with Celsr, Vangl, and a small number of additional membrane and membrane-associated proteins, they also play a central role in tissue polarity/planar cell polarity (PCP) signaling. Targeted mutations in 9 of the 10 mammalian Frizzled genes have revealed their roles in an extraordinarily diverse set of developmental and homeostatic processes, including morphogenetic movements responsible for palate, ventricular septum, ocular furrow, and neural tube closure; survival of thalamic neurons; bone formation; central nervous system (CNS) angiogenesis and blood–brain barrier formation and maintenance; and a wide variety of processes that orient subcellular, cellular, and multicellular structures relative to the body axes. The last group likely reflects the mammalian equivalent of tissue polarity/PCP signaling, as defined in Drosophila, and it includes CNS axon guidance, hair follicle and tongue papilla orientation, and inner ear sensory hair bundle orientation. Frizzled receptors are ubiquitous among multicellular animals and, with other signaling molecules, they very likely evolved to permit the development of the complex tissue architectures that provide multicellular animals with their enormous selective advantage. PMID:26969975

High-resolution orientation and depth of insertion of the voltage-sensing S4 helix of a potassium channel in lipid bilayers.

PubMed

Doherty, Tim; Su, Yongchao; Hong, Mei

2010-08-27

The opening and closing of voltage-gated potassium (Kv) channels are controlled by several conserved Arg residues in the S4 helix of the voltage-sensing domain. The interaction of these positively charged Arg residues with the lipid membrane has been of intense interest for understanding how membrane proteins fold to allow charged residues to insert into lipid bilayers against free-energy barriers. Using solid-state NMR, we have now determined the orientation and insertion depth of the S4 peptide of the KvAP channel in lipid bilayers. Two-dimensional (15)N correlation experiments of macroscopically oriented S4 peptide in phospholipid bilayers revealed a tilt angle of 40 degrees and two possible rotation angles differing by 180 degrees around the helix axis. Remarkably, the tilt angle and one of the two rotation angles are identical to those of the S4 helix in the intact voltage-sensing domain, suggesting that interactions between the S4 segment and other helices of the voltage-sensing domain are not essential for the membrane topology of the S4 helix. (13)C-(31)P distances between the S4 backbone and the lipid (31)P indicate a approximately 9 A local thinning and 2 A average thinning of the DMPC (1,2-dimyristoyl-sn-glycero-3-phosphochloline)/DMPG (1,2-dimyristoyl-sn-glycero-3-phosphatidylglycerol) bilayer, consistent with neutron diffraction data. Moreover, a short distance of 4.6 A from the guanidinium C(zeta) of the second Arg to (31)P indicates the existence of guanidinium phosphate hydrogen bonding and salt bridges. These data suggest that the structure of the Kv gating helix is mainly determined by protein-lipid interactions instead of interhelical protein-protein interactions, and the S4 amino acid sequence encodes sufficient information for the membrane topology of this crucial gating helix. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

Lipid bilayer thickness determines cholesterol's location in model membranes

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

Marquardt, Drew; Heberle, Frederick A.; Greathouse, Denise V.

Cholesterol is an essential biomolecule of animal cell membranes, and an important precursor for the biosynthesis of certain hormones and vitamins. It is also thought to play a key role in cell signaling processes associated with functional plasma membrane microdomains (domains enriched in cholesterol), commonly referred to as rafts. In all of these diverse biological phenomena, the transverse location of cholesterol in the membrane is almost certainly an important structural feature. Using a combination of neutron scattering and solid-state 2H NMR, we have determined the location and orientation of cholesterol in phosphatidylcholine (PC) model membranes having fatty acids of differentmore » lengths and degrees of unsaturation. The data establish that cholesterol reorients rapidly about the bilayer normal in all the membranes studied, but is tilted and forced to span the bilayer midplane in the very thin bilayers. The possibility that cholesterol lies flat in the middle of bilayers, including those made from PC lipids containing polyunsaturated fatty acids (PUFAs), is ruled out. Finally, these results support the notion that hydrophobic thickness is the primary determinant of cholesterol's location in membranes.« less

Lipid bilayer thickness determines cholesterol's location in model membranes

DOE PAGES

Marquardt, Drew; Heberle, Frederick A.; Greathouse, Denise V.; ...

2016-10-11

Cholesterol is an essential biomolecule of animal cell membranes, and an important precursor for the biosynthesis of certain hormones and vitamins. It is also thought to play a key role in cell signaling processes associated with functional plasma membrane microdomains (domains enriched in cholesterol), commonly referred to as rafts. In all of these diverse biological phenomena, the transverse location of cholesterol in the membrane is almost certainly an important structural feature. Using a combination of neutron scattering and solid-state 2H NMR, we have determined the location and orientation of cholesterol in phosphatidylcholine (PC) model membranes having fatty acids of differentmore » lengths and degrees of unsaturation. The data establish that cholesterol reorients rapidly about the bilayer normal in all the membranes studied, but is tilted and forced to span the bilayer midplane in the very thin bilayers. The possibility that cholesterol lies flat in the middle of bilayers, including those made from PC lipids containing polyunsaturated fatty acids (PUFAs), is ruled out. Finally, these results support the notion that hydrophobic thickness is the primary determinant of cholesterol's location in membranes.« less

Robust hydrophobic polyurethane fibrous membranes with tunable porous structure for waterproof and breathable application

NASA Astrophysics Data System (ADS)

Gu, Jiatai; Gu, Haihong; Cao, Jin; Chen, Shaojie; Li, Ni; Xiong, Jie

2018-05-01

In this work, novel nanofibrous membranes with waterproof and breathable (W&B) performance were successfully fabricated by the combination of electrospinning and surface modification technology. This fibrous membranes consisted of polyurethane (PU), NaCl, and fluoroalkylsilane (FAS). Firstly, The fibrous construction and porous structure of fibrous membranes were regulated by tuning the NaCl concentrations in PU solutions. Then, the obtained PU/NaCl fibrous membranes were further modified with fluoroalkylsilane (FAS) to improve hydrophobic property. The synergistic effect of porous structure and hydrophobicity on waterproof and breathable performance was investigated. Furthermore, the mechanical property of fibrous membranes was deeply analysed on the basis of macromolecule orientation and adhesive structure. Benefiting from the optimized porous structure and hydrophobic modification, the resultant fibrous membranes exhibited excellent waterproof (hydrostatic pressure of 1261 Mbar), breathable (water vapor transmission (WVT) rate of 9.06 kg m-2 d-1 and air permeability of 4.8 mm s-1) performance, as well as high tensile strength (breakage stress of 10.4 MPa), suggesting a promising candidate for various applications, especially in protective clothing.

Control of Transmembrane Helix Dynamics by Interfacial Tryptophan Residues.

PubMed

McKay, Matthew J; Martfeld, Ashley N; De Angelis, Anna A; Opella, Stanley J; Greathouse, Denise V; Koeppe, Roger E

2018-06-05

Transmembrane protein domains often contain interfacial aromatic residues, which may play a role in the insertion and stability of membrane helices. Residues such as Trp or Tyr, therefore, are often found situated at the lipid-water interface. We have examined the extent to which the precise radial locations of interfacial Trp residues may influence peptide helix orientation and dynamics. To address these questions, we have modified the GW 5,19 ALP23 (acetyl-GGALW 5 (LA) 6 LW 19 LAGA-[ethanol]amide) model peptide framework to relocate the Trp residues. Peptide orientation and dynamics were analyzed by means of solid-state nuclear magnetic resonance (NMR) spectroscopy to monitor specific 2 H- and 15 N-labeled residues. GW 5,19 ALP23 adopts a defined, tilted orientation within lipid bilayer membranes with minimal evidence of motional averaging of NMR observables, such as 2 H quadrupolar or 15 N- 1 H dipolar splittings. Here, we examine how peptide dynamics are impacted by relocating the interfacial Trp (W) residues on both ends and opposing faces of the helix, for example by a 100° rotation on the helical wheel for positions 4 and 20. In contrast to GW 5,19 ALP23, the modified GW 4,20 ALP23 helix experiences more extensive motional averaging of the NMR observables in several lipid bilayers of different thickness. Individual and combined Gaussian analyses of the 2 H and 15 N NMR signals confirm that the extent of dynamic averaging, particularly rotational "slippage" about the helix axis, is strongly coupled to the radial distribution of the interfacial Trp residues as well as the bilayer thickness. Additional 2 H labels on alanines A3 and A21 reveal partial fraying of the helix ends. Even within the context of partial unwinding, the locations of particular Trp residues around the helix axis are prominent factors for determining transmembrane helix orientation and dynamics within the lipid membrane environment. Copyright © 2018 Biophysical Society. Published by Elsevier Inc. All rights reserved.

Structure of the tandem PX-PH domains of Bem3 from Saccharomyces cerevisiae.

PubMed

Ali, Imtiaz; Eu, Sungmin; Koch, Daniel; Bleimling, Nathalie; Goody, Roger S; Müller, Matthias P

2018-05-01

The structure of the tandem lipid-binding PX and pleckstrin-homology (PH) domains of the Cdc42 GTPase-activating protein Bem3 from Saccharomyces cerevisiae (strain S288c) has been determined to a resolution of 2.2 Å (R work = 21.1%, R free = 23.4%). It shows that the domains adopt a relative orientation that enables them to simultaneously bind to a membrane and suggests possible cooperativity in membrane binding. open access.

Structure of the tandem PX-PH domains of Bem3 from Saccharomyces cerevisiae

PubMed Central

Ali, Imtiaz; Eu, Sungmin; Bleimling, Nathalie

2018-01-01

The structure of the tandem lipid-binding PX and pleckstrin-homology (PH) domains of the Cdc42 GTPase-activating protein Bem3 from Saccharomyces cerevisiae (strain S288c) has been determined to a resolution of 2.2 Å (R work = 21.1%, R free = 23.4%). It shows that the domains adopt a relative orientation that enables them to simultaneously bind to a membrane and suggests possible cooperativity in membrane binding. PMID:29718000

Mineralization of the Sea Urchin Skeleton

NASA Astrophysics Data System (ADS)

Wilt, F.

2001-12-01

The sea urchin possess a calcareous skeleton composed of over 99% magnesian calcite,an enveloping extracellular matrix, and an occluded protein matrix. The most intensively studied skeletal element is the spicule of the embryo. At the 32 cell stage of development a cohort of 4 cells becomes irrevocably dedicated to spicule formation. At the early gastrula stage the descendants of these founder cells form the primary mesenchyme (PMC). The PMCs fuse to form a multinucleated syncytium connected by cytoplasmic cables, and the calcitic skeleton is formed within these cables. Our primary concern is with the cellular and molecular mechanisms that support the formation of the mineralized spicules. The import of calcium into the PMCs results in appearance of intracellular vesicles containing precipitated calcium, which is neither very stable nor birefringent, and could be amorphous. The precipitated calcium is vectorially secreted into an extracellular space. This space is almost completely enclosed by cytoplasmic strands, and the mineral is encased in an extracellular matrix. Proteins destined for the extracellular matrix, and for inclusion in the spicule, are present in the Golgi membranes and in small intracellular vesicles. These vesicles apparently deliver the matrix proteins to the growing spicule. Our current view is that the matrix molecules are much more than a passive armature, but are actively involved in precipitation, secretion, and organization of the mineral phase.

Modeling of annexin A2-Membrane interactions by molecular dynamics simulations.

PubMed

Hakobyan, Davit; Gerke, Volker; Heuer, Andreas

2017-01-01

The annexins are a family of Ca2+-regulated phospholipid binding proteins that are involved in membrane domain organization and membrane trafficking. Although they are widely studied and crystal structures are available for several soluble annexins their mode of membrane association has never been studied at the molecular level. Here we obtained molecular information on the annexin-membrane interaction that could serve as paradigm for the peripheral membrane association of cytosolic proteins by Molecular Dynamics simulations. We analyzed systems containing the monomeric annexin A2 (AnxA2), a membrane with negatively charged phosphatidylserine (POPS) lipids as well as Ca2+ ions. On the atomic level we identify the AnxA2 orientations and the respective residues which display the strongest interaction with Ca2+ ions and the membrane. The simulation results fully agree with earlier experimental findings concerning the positioning of bound Ca2+ ions. Furthermore, we identify for the first time a significant interaction between lysine residues of the protein and POPS lipids that occurs independently of Ca2+ suggesting that AnxA2-membrane interactions can also occur in a low Ca2+ environment. Finally, by varying Ca2+ concentrations and lipid composition in our simulations we observe a calcium-induced negative curvature of the membrane as well as an AnxA2-induced lipid ordering.

The innovative osmotic membrane bioreactor (OMBR) for reuse of wastewater.

PubMed

Cornelissen, E R; Harmsen, D; Beerendonk, E F; Qin, J J; Oo, H; de Korte, K F; Kappelhof, J W M N

2011-01-01

An innovative osmotic membrane bioreactor (OMBR) is currently under development for the reclamation of wastewater, which combines activated sludge treatment and forward osmosis (FO) membrane separation with a RO post-treatment. The research focus is FO membrane fouling and performance using different activated sludge investigated both at laboratory scale (membrane area of 112cm2) and at on-site bench scale (flat sheet membrane area of 0.1 m2). FO performance on laboratory-scale (i) increased with temperature due to a decrease in viscosity and (ii) was independent of the type of activated sludge. Draw solution leakage increased with temperature and varied for different activated sludge. FO performance on bench-scale (i) increased with osmotic driving force, (ii) depended on the membrane orientation due to internal concentration polarization and (iii) was invariant to feed flow decrease and air injection at the feed and draw side. Draw solution leakage could not be evaluated on bench-scale due to experimental limitation. Membrane fouling was not found on laboratory scale and bench-scale, however, partially reversible fouling was found on laboratory scale for FO membranes facing the draw solution. Economic assessment indicated a minimum flux of 15L.m-2 h-1 at 0.5M NaCl for OMBR-RO to be cost effective, depending on the FO membrane price.

[Computer modeling the hydrostatic pressure characteristics of the membrane potential for polymeric membrane, separated non-homogeneous electrolyte solutions].

PubMed

Slezak, Izabella H; Jasik-Slezak, Jolanta; Rogal, Mirosława; Slezak, Andrzej

2006-01-01

On the basis of model equation depending the membrane potential deltapsis, on mechanical pressure difference (deltaP), concentration polarization coefficient (zetas), concentration Rayleigh number (RC) and ratio concentration of solutions separated by membrane (Ch/Cl), the characteristics deltapsis = f(deltaP)zetas,RC,Ch/Cl for steady values of zetas, RC and Ch/Cl in single-membrane system were calculated. In this system neutral and isotropic polymeric membrane oriented in horizontal plane, the non-homogeneous binary electrolytic solutions of various concentrations were separated. Nonhomogeneity of solutions is results from creations of the concentration boundary layers on both sides of the membrane. Calculations were made for the case where on a one side of the membrane aqueous solution of NaCl at steady concentration 10(-3) mol x l(-1) (Cl) was placed and on the other aqueous solutions of NaCl at concentrations from 10(-3) mol x l(-1) to 2 x 10(-2) mol x l(-1) (Ch). Their densities were greater than NaCl solution's at 10(-3) mol x l(-1). It was shown that membrane potential depends on hydrodynamic state of a complex concentration boundary layer-membrane-concentration boundary layer, what is controlled by deltaP, Ch/Cl, RC and zetas.

Theoretical investigation of Lamb wave characteristics in AlN/3C-SiC composite membranes

NASA Astrophysics Data System (ADS)

Lin, Chih-Ming; Chen, Yung-Yu; Pisano, Albert P.

2010-11-01

Cubic silicon carbide (3C-SiC) layer can provide advantages of high frequency and high quality factor for Lamb wave devices due to the superior properties of high acoustic velocity and low acoustic loss. In this study, Lamb wave propagation characteristics in composite membranes consisting of a c-axis oriented aluminum nitride (AlN) film and an epitaxial 3C-SiC (100) layer are investigated by theoretical calculation. The lowest symmetric mode Lamb wave propagating along the [011] direction exhibits a phase velocity higher than 10 000 m/s and an electromechanical coupling coefficient above 2% in the AlN/3C-SiC multilayered membranes.

A facile method to align carbon nanotubes on polymeric membrane substrate

PubMed Central

Zhao, Haiyang; Zhou, Zhijun; Dong, Hang; Zhang, Lin; Chen, Huanlin; Hou, Lian

2013-01-01

The alignment of carbon nanotubes (CNT) is the fundamental requirement to ensure their excellent functions but seems to be desolated in recent years. A facile method, hot-press combined with peel-off (HPPO), is introduced here, through which CNT can be successfully vertically aligned on the polymeric membrane substrate. Shear force and mechanical stretch are proposed to be the main forces to align the tubes perpendicular to the substrate surface during the peel-off process. The alignment of CNT keeps its orientation in a thin hybrid membrane by dip-coating cellulose acetate dope solution. It is expected that the stable alignment of CNT by HPPO would contribute to the realization of its potential applications. PMID:24326297

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

PubMed

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

2017-12-20

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

Spacer geometry and particle deposition in spiral wound membrane feed channels.

PubMed

Radu, A I; van Steen, M S H; Vrouwenvelder, J S; van Loosdrecht, M C M; Picioreanu, C

2014-11-01

Deposition of microspheres mimicking bacterial cells was studied experimentally and with a numerical model in feed spacer membrane channels, as used in spiral wound nanofiltration (NF) and reverse osmosis (RO) membrane systems. In-situ microscopic observations in membrane fouling simulators revealed formation of specific particle deposition patterns for different diamond and ladder feed spacer orientations. A three-dimensional numerical model combining fluid flow with a Lagrangian approach for particle trajectory calculations could describe very well the in-situ observations on particle deposition in flow cells. Feed spacer geometry, positioning and cross-flow velocity sensitively influenced the particle transport and deposition patterns. The deposition patterns were not influenced by permeate production. This combined experimental-modeling approach could be used for feed spacer geometry optimization studies for reduced (bio)fouling. Copyright © 2014 Elsevier Ltd. All rights reserved.

Studying the spatial organization of membrane proteins by means of tritium stratigraphy: bacteriorhodopsin in purple membrane.

PubMed

Shishkov, A V; Ksenofontov, A L; Bogacheva, E N; Kordyukova, L V; Badun, G A; Alekseevsky, A V; Tsetlin, V I; Baratova, L A

2002-05-15

The topography of bacteriorhodopsin (bR) in situ was earlier studied by using the tritium bombardment approach [Eur. J. Biochem. 178 (1988) 123]. Now, having the X-ray crystallography data of bR at atom resolution [Proc. Natl. Acad. Sci. 95 (1998) 11673], we estimated the influence of membrane environment (lipid and protein) on tritium incorporation into amino acid residues forming transmembrane helices. We have determined the tritium flux attenuation coefficients for residues 10-29 of helix A. They turned out to be low (0.04+/-0.02 A(-1)) for residues adjacent to the lipid matrix, and almost fourfold higher (0.15+/-0.05 A(-1)) for those oriented to the neighboring transmembrane helices. We believe that tritium incorporation data could help modeling transmembrane segment arrangement in the membrane.

Molecular insight into the electrostatic membrane surface potential by 14n/31p MAS NMR spectroscopy: nociceptin-lipid association.

PubMed

Lindström, Fredrick; Williamson, Philip T F; Gröbner, Gerhard

2005-05-11

Exploiting naturally abundant (14)N and (31)P nuclei by high-resolution MAS NMR (magic angle spinning nuclear magnetic resonance) provides a molecular view of the electrostatic potential present at the surface of biological model membranes, the electrostatic charge distribution across the membrane interface, and changes that occur upon peptide association. The spectral resolution in (31)P and (14)N MAS NMR spectra is sufficient to probe directly the negatively charged phosphate and positively charged choline segment of the electrostatic P(-)-O-CH(2)-CH(2)-N(+)(CH(3))(3) headgroup dipole of zwitterionic DMPC (dimyristoylphosphatidylcholine) in mixed-lipid systems. The isotropic shifts report on the size of the potential existing at the phosphate and ammonium group within the lipid headgroup while the chemical shielding anisotropy ((31)P) and anisotropic quadrupolar interaction ((14)N) characterize changes in headgroup orientation in response to surface potential. The (31)P/(14)N isotropic chemical shifts for DMPC show opposing systematic changes in response to changing membrane potential, reflecting the size of the electrostatic potential at opposing ends of the P(-)-N(+) dipole. The orientational response of the DMPC lipid headgroup to electrostatic surface variations is visible in the anisotropic features of (14)N and (31)P NMR spectra. These features are analyzed in terms of a modified "molecular voltmeter" model, with changes in dynamic averaging reflecting the tilt of the C(beta)-N(+)(CH)(3) choline and PO(4)(-) segment. These properties have been exploited to characterize the changes in surface potential upon the binding of nociceptin to negatively charged membranes, a process assumed to proceed its agonistic binding to its opoid G-protein coupled receptor.

Sensitivity enhancement and contrasting information provided by free radicals in oriented-sample NMR of bicelle-reconstituted membrane proteins.

PubMed

Tesch, Deanna M; Nevzorov, Alexander A

2014-02-01

Elucidating structure and topology of membrane proteins (MPs) is essential for unveiling functionality of these important biological constituents. Oriented-sample solid-state NMR (OS-NMR) is capable of providing such information on MPs under nearly physiological conditions. However, two dimensional OS-NMR experiments can take several days to complete due to long longitudinal relaxation times combined with the large number of scans to achieve sufficient signal sensitivity in biological samples. Here, free radicals 5-DOXYL stearic acid, TEMPOL, and CAT-1 were added to uniformly (15)N-labeled Pf1 coat protein reconstituted in DMPC/DHPC bicelles, and their effect on the longitudinal relaxation times (T1Z) was investigated. The dramatically shortened T1Z's allowed for the signal gain per unit time to be used for either: (i) up to a threefold reduction of the total experimental time at 99% magnetization recovery or (ii) obtaining up to 74% signal enhancement between the control and radical samples during constant experimental time at "optimal" relaxation delays. In addition, through OS-NMR and high-field EPR studies, free radicals were able to provide positional constraints in the bicelle system, which provide a description of the location of each residue in Pf1 coat protein within the bicellar membranes. This information can be useful in the determination of oligomerization states and immersion depths of larger membrane proteins. Copyright © 2013 Elsevier Inc. All rights reserved.

A transmission and scanning electron microscopic study of the saccule in five species of catfishes.

PubMed

Jenkins, D B

1979-01-01

The sacculi of five species of catfishes were studied by transmission and scanning electron microscopy. In four species, the sagitta exhibited a multifluted anterior part and a tapered posterior part; in Corydoras aeneus, however, the fluted part was absent, and a vertical component extended dorsally to terminate near the opening of the transverse canal. In all species, the otoliths had a laminar structure. An otolithic membrane was present, and hair cell bundles projected into cavities on the macular surface of the membrane. Attachments of the otolithic membrane to the neuroepithelium included short extensions of the membrane to the tallest components of the hair cell bundles of the peripheral cells and more delicate connections to the kinocilium and taller stereocilia of central cells; in addition, attachments to the microvilli of supporting cells were present. In both hair cells and supporting cells single microtubules and bundles of microtubules were present; the bundles had an orderly arrangement and were associated with cytoplasmic densities surrounding the desmosomes. The hair cells were innervated by both afferent and efferent nerve endings. Studies of the polarization of the hair cells in all species (except C. aeneus) showed that there was a single longitudinal axis that divided dorsally polarized cells from those oriented ventrally. In Doras spinosissimus and Bunocephalus bicolor, an additional line of polarization was evident in a small area in the anterior part of the macula; therefore, in these forms there was a double bipolar orientation.

Subsurface microscopy of interconnect layers of an integrated circuit.

PubMed

Köklü, F Hakan; Unlü, M Selim

2010-01-15

We apply the NA-increasing lens technique to confocal and wide-field backside microscopy of integrated circuits. We demonstrate 325 nm (lambda(0)/4) lateral spatial resolution while imaging metal structures located inside the interconnect layer of an integrated circuit. Vectorial field calculations are presented justifying our findings.

Cryptographic Properties of the Hidden Weighted Bit Function

DTIC Science & Technology

2013-12-23

valid OMB control number. 1. REPORT DATE 23 DEC 2013 2. REPORT TYPE 3. DATES COVERED 00-00-2013 to 00-00-2013 4. TITLE AND SUBTITLE...K. Feng, An Infinite Class of Balanced Vectorial Boolean Functions with Optimum Algebraic Immunity and Good Nonlinearity, in: IWCC 2009, In: LNCS

Imaging trypsin activity through changes in the orientation of liquid crystals coupled to the interactions between a polyelectrolyte and a phospholipid layer.

PubMed

Hu, Qiong-Zheng; Jang, Chang-Hyun

2012-03-01

In this study, we developed a new type of liquid crystal (LC)-based sensor for the real-time and label-free monitoring of enzymatic activity through changes in the orientation of LCs coupled to the interactions between polyelectrolyte and phospholipid. The LCs changed from dark to bright after an aqueous solution of poly-l-lysine (PLL) was transferred onto a self-assembled monolayer of the phospholipid, dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), at the aqueous/LC interface. Interactions between the positively charged PLL and the negatively charged DOPG drove the reorganization of the phospholipid membrane, which induced an orientational transition in the LCs from a homeotropic to planar state. Since the serine endopeptidase trypsin can enzymatically catalyze the hydrolysis of PLL, the dark-to-bright shift in the optical response was not observed after transferring a mixed solution of PLL and trypsin onto the DOPG-decorated LC interface, indicating that no orientational transitions in the LCs occurred. However, the optical response from dark to bright was observed when the mixture in the optical cell was replaced by an aqueous solution of PLL. Control experiments with trypsin or an aqueous mixture of PLL and deactivated trypsin further confirmed the feasibility of this approach. The detection limit of trypsin was determined to be ~1 μg/mL. This approach holds great promise for use in the development of LC-based sensors for the detection of enzymatic reactions in cases where the biological polyelectrolyte substrates of enzymes could disrupt the organization of the membrane and induce orientational transitions of LCs at the aqueous/LC interface. © 2012 American Chemical Society

Dimeric arrangement and structure of the membrane-bound acetylcholine receptor studied by electron microscopy.

PubMed Central

Zingsheim, H P; Neugebauer, D C; Frank, J; Hänicke, W; Barrantes, F J

1982-01-01

The acetylcholine receptor protein (AChR) from the electric organ of Torpedo marmorata is studied in its membrane-bound form by electron microscopy and single-particle image averaging. About half the molecule protrudes from the membrane surface by approximately 5 nm. The low-resolution 3-D structure of this hydrated portion, including its handedness, can be deduced from averaged axial and lateral projections and from freeze-etched membrane surfaces. In native membrane fragments, a dimeric form of the AChR is observed and the relative orientation of the AChR monomers within the dimer is established. The dimers disappear upon disulfide reduction of the membrane preparations, whereas the average axial projections of the AChR monomer remain unaffected. Since the existence of disulfide bonds linking AChR monomers between their respective delta-subunits is well documented, the approximate position of the delta-subunit within the low-resolution structure of the AChR molecule can be deduced from the structure of the dimers. Images Fig. 1. Fig. 2. Fig. 3. PMID:7188351

Detergent Optimized Membrane Protein Reconstitution in Liposomes for Solid State NMR

PubMed Central

2015-01-01

For small helical membrane proteins, their structures are highly sensitive to their environment, and solid state NMR is a structural technique that can characterize these membrane proteins in native-like lipid bilayers and proteoliposomes. To date, a systematic method by which to evaluate the effect of the solubilizing detergent on proteoliposome preparations for solid state NMR of membrane proteins has not been presented in the literature. A set of experiments are presented aimed at determining the conditions most amenable to dialysis mediated reconstitution sample preparation. A membrane protein from M. tuberculosis is used to illustrate the method. The results show that a detergent that stabilizes the most protein is not always ideal and sometimes cannot be removed by dialysis. By focusing on the lipid and protein binding properties of the detergent, proteoliposome preparations can be readily produced, which provide double the signal-to-noise ratios for both the oriented sample and magic angle spinning solid state NMR. The method will allow more membrane protein drug targets to be structurally characterized in lipid bilayer environments. PMID:24665863

The cuttlefish Sepia officinalis (Sepiidae, Cephalopoda) constructs cuttlebone from a liquid-crystal precursor

PubMed Central

Checa, Antonio G.; Cartwright, Julyan H. E.; Sánchez-Almazo, Isabel; Andrade, José P.; Ruiz-Raya, Francisco

2015-01-01

Cuttlebone, the sophisticated buoyancy device of cuttlefish, is made of extensive superposed chambers that have a complex internal arrangement of calcified pillars and organic membranes. It has not been clear how this structure is assembled. We find that the membranes result from a myriad of minor membranes initially filling the whole chamber, made of nanofibres evenly oriented within each membrane and slightly rotated with respect to those of adjacent membranes, producing a helical arrangement. We propose that the organism secretes a chitin–protein complex, which self-organizes layer-by-layer as a cholesteric liquid crystal, whereas the pillars are made by viscous fingering. The liquid crystallization mechanism permits us to homologize the elements of the cuttlebone with those of other coleoids and with the nacreous septa and the shells of nautiloids. These results challenge our view of this ultra-light natural material possessing desirable mechanical, structural and biological properties, suggesting that two self-organizing physical principles suffice to understand its formation. PMID:26086668

MFP1 is a thylakoid-associated, nucleoid-binding protein with a coiled-coil structure

PubMed Central

Jeong, Sun Yong; Rose, Annkatrin; Meier, Iris

2003-01-01

Plastid DNA, like bacterial and mitochondrial DNA, is organized into protein–DNA complexes called nucleoids. Plastid nucleoids are believed to be associated with the inner envelope in developing plastids and the thylakoid membranes in mature chloroplasts, but the mechanism for this re-localization is unknown. Here, we present the further characterization of the coiled-coil DNA-binding protein MFP1 as a protein associated with nucleoids and with the thylakoid membranes in mature chloroplasts. MFP1 is located in plastids in both suspension culture cells and leaves and is attached to the thylakoid membranes with its C-terminal DNA-binding domain oriented towards the stroma. It has a major DNA-binding activity in mature Arabidopsis chloroplasts and binds to all tested chloroplast DNA fragments without detectable sequence specificity. Its expression is tightly correlated with the accumulation of thylakoid membranes. Importantly, it is associated in vivo with nucleoids, suggesting a function for MFP1 at the interface between chloroplast nucleoids and the developing thylakoid membrane system. PMID:12930969

Lateral microheterogeneity of diphenylhexatriene-labeled choline phospholipids in the erythrocyte ghost membrane as determined by time-resolved fluorescence spectroscopy.

PubMed

Prenner, E; Sommer, A; Maurer, N; Glatter, O; Gorges, R; Paltauf, F; Hermetter, A

2000-04-01

Choline phospholipids are the major constituents of the outer layer of the erythrocyte membrane. To investigate their lateral membrane organization we determined the fluorescence lifetime properties of diphenylhexatriene analogues of phosphatidylcholine, choline plasmalogen, (the respective enolether derivative), and sphingomyelin inserted into the outer layer of hemoglobin-free ghosts. Fluorescence lifetimes were recorded by time-resolved phase and modulation fluorometry and analyzed in terms of Continuous Lorentzian distributions. To assess the influence of membrane proteins on the fluorescence lifetime of the labeled lipids in the biomembrane, lipid vesicles were used as controls. In general, the lifetime distributions in the ghost membranes are broad compared to vesicles. Phosphatidylcholine and sphingomyelin exhibit very similar lifetime distributions in contrast to an increased plasmalogen lifetime heterogeneity in both systems. Orientational effects of side chain mobilities on the observed lifetimes can be excluded. Fluorescence anisotropies revealed identical values for all three labeled phospholipids in the biomembrane.

Freely suspended nanocomposite membranes as highly sensitive sensors

NASA Astrophysics Data System (ADS)

Jiang, Chaoyang; Markutsya, Sergiy; Pikus, Yuri; Tsukruk, Vladimir V.

2004-10-01

Highly sensitive sensor arrays are in high demand for prospective applications in remote sensing and imaging. Measuring microscopic deflections of compliant micromembranes and cantilevers is developing into one of the most versatile approaches for thermal, acoustic and chemical sensing. Here, we report on an innovative fabrication of compliant nanocomposite membranes with nanoscale thickness showing extraordinary sensitivity and dynamic range, which makes them candidates for a new generation of membrane-based sensor arrays. These nanomembranes with a thickness of 25-70 nm, which can be freely suspended over large (hundred micrometres) openings are fabricated with molecular precision by time-efficient, spin-assisted layer-by-layer assembly. They are designed as multilayered molecular composites made of a combination of polymeric monolayers and a metal nanoparticle intralayer. We demonstrate that these nanocomposite membranes possess unparalleled sensitivity and a unique autorecovering ability. The membrane nanostructure that is responsible for these outstanding properties combines multilayered polymer/nanoparticle organization, high polymer-chain orientation, and a pre-stretched state.

[Computer modeling the dependences of the membrane potential for polymeric membrane separated non-homogeneous electrolyte solutions on concentration Rayleigh number].

PubMed

Slezak, Izabella H; Jasik-Slezak, Jolanta; Bilewicz-Wyrozumska, Teresa; Slezak, Andrzej

2006-01-01

On the basis of model equation describing the membrane potential delta psi(s) on concentration Rayleigh number (R(C)), mechanical pressure difference (deltaP), concentration polarization coefficient (zeta s) and ratio concentration of solutions separated by membrane (Ch/Cl), the characteristics delta psi(s) = f(Rc)(delta P, zeta s, Ch/Cl) for steady values of zeta s, R(C) and Ch/Cl in single-membrane system were calculated. In this system neutral and isotropic polymeric membrane oriented in horizontal plane, the non-homogeneous binary electrolytic solutions of various concentrations were separated. Nonhomogeneity of solutions is results from creations of the concentration boundary layers on both sides of the membrane. Calculations were made for the case where on a one side of the membrane aqueous solution of NaCl at steady concentration 10(-3) mol x l(-1) (Cl) was placed and on the other aqueous solutions of NaCl at concentrations from 10(-3) mol x l(-1) to 2 x 10(-2) mol x l(-1) (Ch). Their densities were greater than NaCl solution's at 10(-3) mol x l(-1). It was shown that membrane potential depends on hydrodynamic state of a complex concentration boundary layer-membrane-concentration boundary layer, what is controlled by deltaP, Ch/Cl, Rc and Zeta(s).

Effect of Melatonin and Cholesterol on the Structure of DOPC and DPPC Membranes

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

Drolle, E; Kucerka, Norbert; Hoopes, M I

The cell membrane plays an important role in the molecular mechanism of amyloid toxicity associated with Alzheimer's disease. The membrane's chemical composition and the incorporation of small molecules, such as melatonin and cholesterol, can alter its structure and physical properties, thereby affecting its interaction with amyloid peptides. Both melatonin and cholesterol have been recently linked to amyloid toxicity. Melatonin has been shown to have a protective role against amyloid toxicity. However, the underlying molecular mechanism of this protection is still not well understood, and cholesterol's role remains controversial. We used small-angle neutron diffraction (SAND) from oriented lipid multi-layers, small-angle neutronmore » scattering (SANS) from unilamellar vesicles experiments andMolecular Dynamics (MD) simulations to elucidate non-specific interactions of melatonin and cholesterol with 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) and 1,2-dipalmitoyl-snglycero-3-phosphocholine (DPPC) model membranes. We conclude that melatonin decreases the thickness of both model membranes by disordering the lipid hydrocarbon chains, thus increasing membrane fluidity. This result is in stark contrast to the much accepted ordering effect induced by cholesterol, which causes membranes to thicken.« less

Ras trafficking, localization and compartmentalized signalling

PubMed Central

Prior, Ian A.; Hancock, John F.

2012-01-01

Ras proteins are proto-oncogenes that are frequently mutated in human cancers. Three closely related isoforms, HRAS, KRAS and NRAS, are expressed in all cells and have overlapping but distinctive functions. Recent work has revealed how differences between the Ras isoforms in their trafficking, localization and protein-membrane orientation enable signalling specificity to be determined. We review the various strategies used to characterize compartmentalized Ras localization and signalling. Localization is an important contextual modifier of signalling networks and insights from the Ras system are of widespread relevance for researchers interested in signalling initiated from membranes. PMID:21924373

Amyloplasts and Vacuolar Membrane Dynamics in the Living Graviperceptive Cell of the Arabidopsis Inflorescence StemW⃞

PubMed Central

Saito, Chieko; Morita, Miyo T.; Kato, Takehide; Tasaka, Masao

2005-01-01

We developed an adequate method for the in vivo analysis of organelle dynamics in the gravity-perceptive cell (endodermis) of the Arabidopsis thaliana inflorescence stem, revealing behavior of amyloplasts and vacuolar membranes in those cells. Amyloplasts in the endodermis showed saltatory movements even before gravistimulation by reorientation, and these movements were confirmed as microfilament dependent. From our quantitative analysis in the wild type, the gravity-oriented movement of amyloplasts mainly occurred during 0 to 3 min after gravistimulation by reorientation, supporting findings from our previous physiological study. Even after microfilament disruption, the gravity-oriented movement of amyloplasts remained. By contrast, in zig/sgr4 mutants, where a SNARE molecule functioning in vacuole biogenesis has been disrupted, the movement of amyloplasts in the endodermis is severely restricted both before and after gravistimulation by reorientation. Here, we describe vacuolar membrane behavior in these cells in the wild-type, actin filament–disrupted, and zig/sgr4 mutants and discuss its putatively important features for the perception of gravity. We also discuss the data on the two kinds of movements of amyloplasts that may play an important role in gravitropism: (1) the leading edge amyloplasts and (2) the en mass movement of amyloplasts. PMID:15689424

Electrochemical mechanism of tin membrane electrodeposition under ultrasonic waves.

PubMed

Nan, Tianxiang; Yang, Jianguang; Chen, Bing

2018-04-01

Tin was electrodeposited from chloride solutions using a membrane cell under ultrasonic waves. Cyclic voltammetry (CV), linear sweep voltammetry (LSV), chronoamperometry (CHR), and chronopotentiometry were applied to investigate the electrochemical mechanism of tin electrodeposition under ultrasonic field. Chronoamperometry curves showed that the initial process of tin electrodeposition followed the diffusion controlled three-dimensional nucleation and grain growth mechanism. The analysis of the cyclic voltammetry and linear sweep voltammetry diagrams showed that the application of ultrasound can change the tin membrane electro-deposition reaction from diffusion to electrochemical control, and the optimum parameters for tin electrodeposition were H + concentration 3.5 mol·L -1 , temperature 35 °C and ultrasonic power 100 W. The coupling ultrasonic field played a role in refining the grain in this process. The growth of tin crystals showed no orientation preferential, and the tin deposition showed a tendency to form a regular network structure after ultrasonic coupling. While in the absence of ultrasonic coupling, the growth of tin crystals has a high preferential orientation, and the tin deposition showed a tendency to form tin whiskers. Ultrasonic coupling was more favorable for obtaining a more compact and smoother cathode tin layer. Copyright © 2017 Elsevier B.V. All rights reserved.

Theoretical vibrational sum-frequency generation spectroscopy of water near lipid and surfactant monolayer interfaces.

PubMed

Roy, S; Gruenbaum, S M; Skinner, J L

2014-11-14

Understanding the structure of water near cell membranes is crucial for characterizing water-mediated events such as molecular transport. To obtain structural information of water near a membrane, it is useful to have a surface-selective technique that can probe only interfacial water molecules. One such technique is vibrational sum-frequency generation (VSFG) spectroscopy. As model systems for studying membrane headgroup/water interactions, in this paper we consider lipid and surfactant monolayers on water. We adopt a theoretical approach combining molecular dynamics simulations and phase-sensitive VSFG to investigate water structure near these interfaces. Our simulated spectra are in qualitative agreement with experiments and reveal orientational ordering of interfacial water molecules near cationic, anionic, and zwitterionic interfaces. OH bonds of water molecules point toward an anionic interface leading to a positive VSFG peak, whereas the water hydrogen atoms point away from a cationic interface leading to a negative VSFG peak. Coexistence of these two interfacial water species is observed near interfaces between water and mixtures of cationic and anionic lipids, as indicated by the presence of both negative and positive peaks in their VSFG spectra. In the case of a zwitterionic interface, OH orientation is toward the interface on the average, resulting in a positive VSFG peak.

Actin-based gravity-sensing mechanisms in unicellular plant model systems

NASA Astrophysics Data System (ADS)

Braun, Markus; Limbach, Christoph

2005-08-01

Considerable progress has been made in the understanding of the molecular and cellular mechanisms underlying gravity sensing and gravity-oriented polarized growth in single-celled rhizoids and protonemata of the characean algae. It is well known that the actin cytoskeleton plays a key role in these processes. Numerous actin-binding proteins control apical actin polymerization and the dynamic remodeling of the actin arrangement. An actomyosin-based system mediates the delivery and incorporation of secretory vesicles at the growing tip and coordinates the tip-high gradient of cytoplasmic free calcium which is required for local exocytosis. Additionally, the actomyosin system precisely controls the position of statoliths and, upon a change in orientation relative to the gravity vector, directs sedimenting statoliths to the confined graviperception sites of the plasma membrane where gravitropic signalling is initiated. The upward growth response of protonemata is preceded by an actin-dependent relocalization of the Ca2+-gradient to the upper flank. The downward growth response of rhizoids, however, is caused by differential growth of the opposite flankes due to a local reduction of cytoplasmic free calcium limited to the plasma membrane area where statoliths are sedimented. Thus, constant actin polymerization in the growing tip and the spatiotemporal control of actin remodeling are essential for gravity sensing and gravity-oriented polarized growth of characean rhizoids and protonemata.

The assembly of cellulose microfibrils in Valonia macrophysa Kütz.

PubMed

Itoh, T; Brown, R M

1984-03-01

The assembly of cellulose microfibrils was investigated in artificially induced protoplasts of the alga, Valonia macrophysa (Siphonocladales). Primary-wall microfibrills, formed within 72 h of protoplast induction, are randomly oriented. Secondary-wall lamellae, which are produced within 96 h after protoplast induction, have more than three orientations of highly ordered microfibrils. The innermost, recently deposited micofibrils are not parallel with the cortical microtubules, thus indicating a more indirect role of microtubules in the orientation of microfibrils. Fine filamentous structures with a periodicity of 5.0-5.5 nm and the dimensions of actin were observed adjacent to the plasma membrane. Linear cellulose-terminal synthesizing complexes (TCs) consisting of three rows, each with 30-40 particles, were observed not only on the E fracture (EF) but also on P fracture (PF) faces of the plasma membrane. The TC appears to span both faces of the bimolecular leaflet. The average length of the TC is 350 nm, and the number of TCs per unit area during primary-wall synthesis is 1 per μm(2). Neither paired TCs nor granule bands characteristic of Oocystis were observed. Changes in TC structure and distribution during the conversion from primary- to secondary-wall formation have been described. Cellulose microfibril assembly in Valonia is discussed in relation to the process among other eukaryotic systems.

Generic instabilities in a fluid membrane coupled to a thin layer of ordered active polar fluid.

PubMed

Sarkar, Niladri; Basu, Abhik

2013-08-01

We develop an effective two-dimensional coarse-grained description for the coupled system of a planar fluid membrane anchored to a thin layer of polar ordered active fluid below. The macroscopic orientation of the active fluid layer is assumed to be perpendicular to the attached membrane. We demonstrate that activity or nonequilibrium drive of the active fluid makes such a system generically linearly unstable for either signature of a model parameter [Formula: see text] [Formula: see text] that characterises the strength of activity. Depending upon boundary conditions and within a range of the model parameters, underdamped propagating waves may be present in our model. We discuss the phenomenological significance of our results.

Hydrophobically stabilized open state for the lateral gate of the Sec translocon

PubMed Central

Zhang, Bin; Miller, Thomas F.

2010-01-01

The Sec translocon is a central component of cellular pathways for protein translocation and membrane integration. Using both atomistic and coarse-grained molecular simulations, we investigate the conformational landscape of the translocon and explore the role of peptide substrates in the regulation of the translocation and integration pathways. Inclusion of a hydrophobic peptide substrate in the translocon stabilizes the opening of the lateral gate for membrane integration, whereas a hydrophilic peptide substrate favors the closed lateral gate conformation. The relative orientation of the plug moiety and a peptide substrate within the translocon channel is similarly dependent on whether the substrate is hydrophobic or hydrophilic in character, and the energetics of the translocon lateral gate opening in the presence of a peptide substrate is governed by the energetics of the peptide interface with the membrane. Implications of these results for the regulation of Sec-mediated pathways for protein translocation vs. membrane integration are discussed. PMID:20203009

Thin Polymer Films with Continuous Vertically Aligned 1 nm Pores Fabricated by Soft Confinement

DOE PAGES

Feng, Xunda; Nejati, Siamak; Cowan, Matthew G.; ...

2015-12-03

Membrane separations are critically important in areas ranging from health care and analytical chemistry to bioprocessing and water purification. An ideal nanoporous membrane would consist of a thin film with physically continuous and vertically aligned nanopores and would display a narrow distribution of pore sizes. However, the current state of the art departs considerably from this ideal and is beset by intrinsic trade-offs between permeability and selectivity. We demonstrate an effective and scalable method to fabricate polymer films with ideal membrane morphologies consisting of submicron thickness films with physically continuous and vertically aligned 1 nm pores. The approach is basedmore » on soft confinement to control the orientation of a cross-linkable mesophase in which the pores are produced by self-assembly. The scalability, exceptional ease of fabrication, and potential to create a new class of nanofiltration membranes stand out as compelling aspects.« less

Peptide-Lipid Interactions: Experiments and Applications

PubMed Central

Galdiero, Stefania; Falanga, Annarita; Cantisani, Marco; Vitiello, Mariateresa; Morelli, Giancarlo; Galdiero, Massimiliano

2013-01-01

The interactions between peptides and lipids are of fundamental importance in the functioning of numerous membrane-mediated cellular processes including antimicrobial peptide action, hormone-receptor interactions, drug bioavailability across the blood-brain barrier and viral fusion processes. Moreover, a major goal of modern biotechnology is obtaining new potent pharmaceutical agents whose biological action is dependent on the binding of peptides to lipid-bilayers. Several issues need to be addressed such as secondary structure, orientation, oligomerization and localization inside the membrane. At the same time, the structural effects which the peptides cause on the lipid bilayer are important for the interactions and need to be elucidated. The structural characterization of membrane active peptides in membranes is a harsh experimental challenge. It is in fact accepted that no single experimental technique can give a complete structural picture of the interaction, but rather a combination of different techniques is necessary. PMID:24036440

Differential Membrane Dipolar Orientation Induced by Acute and Chronic Cholesterol Depletion.

PubMed

Sarkar, Parijat; Chakraborty, Hirak; Chattopadhyay, Amitabha

2017-06-30

Cholesterol plays a crucial role in cell membrane organization, dynamics and function. Depletion of cholesterol represents a popular approach to explore cholesterol-sensitivity of membrane proteins. An emerging body of literature shows that the consequence of membrane cholesterol depletion often depends on the actual process (acute or chronic), although the molecular mechanism underlying the difference is not clear. Acute depletion, using cyclodextrin-type carriers, is faster relative to chronic depletion, in which inhibitors of cholesterol biosynthesis are used. With the overall goal of addressing molecular differences underlying these processes, we monitored membrane dipole potential under conditions of acute and chronic cholesterol depletion in CHO-K1 cells, using a voltage-sensitive fluorescent dye in dual wavelength ratiometric mode. Our results show that the observed membrane dipole potential exhibits difference under acute and chronic cholesterol depletion conditions, even when cholesterol content was identical. To the best of our knowledge, these results provide, for the first time, molecular insight highlighting differences in dipolar reorganization in these processes. A comprehensive understanding of processes in which membrane cholesterol gets modulated would provide novel insight in its interaction with membrane proteins and receptors, thereby allowing us to understand the role of cholesterol in cellular physiology associated with health and disease.

New Insights from Sum Frequency Generation Vibrational Spectroscopy into the Interactions of Islet Amyloid Polypeptides with Lipid Membranes

PubMed Central

Wang, Zhuguang; Batista, Victor S.; Yan, Elsa C. Y.

2016-01-01

Studies of amyloid polypeptides on membrane surfaces have gained increasing attention in recent years. Several studies have revealed that membranes can catalyze protein aggregation and that the early products of amyloid aggregation can disrupt membrane integrity, increasing water permeability and inducing ion cytotoxicity. Nonetheless, probing aggregation of amyloid proteins on membrane surfaces is challenging. Surface-specific methods are required to discriminate contributions of aggregates at the membrane interface from those in the bulk phase and to characterize protein secondary structures in situ and in real time without the use of perturbing spectroscopic labels. Here, we review the most recent applications of sum frequency generation (SFG) vibrational spectroscopy applied in conjunction with computational modeling techniques, a joint experimental and computational methodology that has provided valuable insights into the aggregation of islet amyloid polypeptide (IAPP) on membrane surfaces. These applications show that SFG can provide detailed information about structures, kinetics, and orientation of IAPP during interfacial aggregation, relevant to the molecular mechanisms of type II diabetes. These recent advances demonstrate the promise of SFG as a new approach for studying amyloid diseases at the molecular level and for the rational drug design targeting early aggregation products on membrane surfaces. PMID:26697504

Molecular organization, localization and orientation of antifungal antibiotic amphotericin B in a single lipid bilayer

PubMed Central

Grudzinski, Wojciech; Sagan, Joanna; Welc, Renata; Luchowski, Rafal; Gruszecki, Wieslaw I.

2016-01-01

Amphotericin B is a popular antifungal antibiotic, a gold standard in treatment of systemic mycotic infections, due to its high effectiveness. On the other hand, applicability of the drug is limited by its considerable toxicity to patients. Biomembranes are a primary target of physiological activity of amphotericin B and both the pharmacologically desired and toxic side effects of the drug relay on its molecular organization in the lipid phase. In the present work, molecular organization, localization and orientation of amphotericin B, in a single lipid bilayer system, was analysed simultaneously, thanks to application of a confocal fluorescence lifetime imaging microscopy of giant unilamellar vesicles. The results show that the presence of sterols, in the lipid phase, promotes formation of supramolecular structures of amphotericin B and their penetration into the membrane hydrophobic core. The fact that such an effect is substantially less pronounced in the case of cholesterol than ergosterol, the sterol of fungal membranes, provides molecular insight into the selectivity of the drug. PMID:27620838

The influence of physiological status on age prediction of Anopheles arabiensis using near infra-red spectroscopy

USDA-ARS?s Scientific Manuscript database

Determining the age of malaria vectors is essential for evaluating the impact of interventions that reduce the survival of wild mosquito populations and for estimating changes in vectorial capacity. Near infra-red spectroscopy (NIRS) is a simple and non-destructive method that has been used to deter...

Concatenations of the Hidden Weighted Bit Function and Their Cryptographic Properties

DTIC Science & Technology

2014-01-01

a collection of information if it does not display a currently valid OMB control number. 1. REPORT DATE 2014 2. REPORT TYPE 3. DATES COVERED 00...nonlinearity, in Advances in Cryptology – ASIACRYPT 2008 , Springer-Verlag, 2008, 425–440. [6] C. Carlet and K. Feng, An infinite class of balanced vectorial

A Construction of Boolean Functions with Good Cryptographic Properties

DTIC Science & Technology

2014-01-01

be subject to a penalty for failing to comply with a collection of information if it does not display a currently valid OMB control number. 1. REPORT...2008, LNCS 5350, Springer–Verlag, 2008, pp. 425–440. [10] C. Carlet and K. Feng, “An Infinite Class of Balanced Vectorial Boolean Functions with Optimum

New Vectorial Propulsion System and Trajectory Control Designs for Improved AUV Mission Autonomy.

PubMed

Masmitja, Ivan; Gonzalez, Julian; Galarza, Cesar; Gomariz, Spartacus; Aguzzi, Jacopo; Del Rio, Joaquin

2018-04-17

Autonomous Underwater Vehicles (AUV) are proving to be a promising platform design for multidisciplinary autonomous operability with a wide range of applications in marine ecology and geoscience. Here, two novel contributions towards increasing the autonomous navigation capability of a new AUV prototype (the Guanay II) as a mix between a propelled vehicle and a glider are presented. Firstly, a vectorial propulsion system has been designed to provide full vehicle maneuverability in both horizontal and vertical planes. Furthermore, two controllers have been designed, based on fuzzy controls, to provide the vehicle with autonomous navigation capabilities. Due to the decoupled system propriety, the controllers in the horizontal plane have been designed separately from the vertical plane. This class of non-linear controllers has been used to interpret linguistic laws into different zones of functionality. This method provided good performance, used as interpolation between different rules or linear controls. Both improvements have been validated through simulations and field tests, displaying good performance results. Finally, the conclusion of this work is that the Guanay II AUV has a solid controller to perform autonomous navigation and carry out vertical immersions.

Indoor-breeding of Aedes albopictus in northern peninsular Malaysia and its potential epidemiological implications.

PubMed

Dieng, Hamady; Saifur, Rahman G M; Hassan, Ahmad Abu; Salmah, M R Che; Boots, Michael; Satho, Tomomitsu; Jaal, Zairi; AbuBakar, Sazaly

2010-07-27

The mosquito Ae. albopictus is usually adapted to the peri-domestic environment and typically breeds outdoors. However, we observed its larvae in most containers within homes in northern peninsular Malaysia. To anticipate the epidemiological implications of this indoor-breeding, we assessed some fitness traits affecting vectorial capacity during colonization process. Specifically, we examined whether Ae. albopictus exhibits increased survival, gonotrophic activity and fecundity due to the potential increase in blood feeding opportunities. In a series of experiments involving outdoors and indoors breeding populations, we found that Ae. albopictus lives longer in the indoor environment. We also observed increased nighttime biting activity and lifetime fecundity in indoor/domestic adapted females, although they were similar to recently colonized females in body size. Taken together these data suggest that accommodation of Ae. albopictus to indoor/domestic environment may increase its lifespan, blood feeding success, nuisance and thus vectorial capacity (both in terms of increased vector-host contacts and vector population density). These changes in the breeding behavior of Ae. albopictus, a potential vector of several human pathogens including dengue viruses, require special attention.

A Spectral-SAR Model for the Anionic-Cationic Interaction in Ionic Liquids: Application to Vibrio fischeri Ecotoxicity

PubMed Central

Lacrămă, Ana-Maria; Putz, Mihai V.; Ostafe, Vasile

2007-01-01

Within the recently launched the spectral-structure activity relationship (S-SAR) analysis, the vectorial anionic-cationic model of a generic ionic liquid is proposed, along with the associated algebraic correlation factor in terms of the measured and predicted activity norms. The reliability of the present scheme is tested by assessing the Hansch factors, i.e. lipophylicity, polarizability and total energy, to predict the ecotoxicity endpoints of wide types of ionic liquids with ammonium, pyridinium, phosphonium, choline and imidazolium cations on the aquatic bacteria Vibrio fischeri. The results, while confirming the cationic dominant influence when only lipophylicity is considered, demonstrate that the anionic effect dominates all other more specific interactions. It was also proved that the S-SAR vectorial model predicts considerably higher activity for the ionic liquids than for its anionic and cationic subsystems separately, in all considered cases. Moreover, through applying the least norm-correlation path principle, the complete toxicological hierarchies are presented, unfolding the ecological rules of combined cationic and anionic influences in ionic liquid toxicity.

Invasiveness of Aedes aegypti and Aedes albopictus and Vectorial Capacity for Chikungunya Virus.

PubMed

Lounibos, Leon Philip; Kramer, Laura D

2016-12-15

In this review, we highlight biological characteristics of Aedes aegypti and Aedes albopictus, 2 invasive mosquito species and primary vectors of chikungunya virus (CHIKV), that set the tone of these species' invasiveness, vector competence, and vectorial capacity (VC). The invasiveness of both species, as well as their public health threats as vectors, is enhanced by preference for human blood. Vector competence, characterized by the efficiency of an ingested arbovirus to replicate and become infectious in the mosquito, depends largely on vector and virus genetics, and most A. aegypti and A. albopictus populations thus far tested confer vector competence for CHIKV. VC, an entomological analog of the pathogen's basic reproductive rate (R 0 ), is epidemiologically more important than vector competence but less frequently measured, owing to challenges in obtaining valid estimates of parameters such as vector survivorship and host feeding rates. Understanding the complexities of these factors will be pivotal in curbing CHIKV transmission. © The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.

Electromagnetic frozen waves with radial, azimuthal, linear, circular, and elliptical polarizations

NASA Astrophysics Data System (ADS)

Corato-Zanarella, Mateus; Zamboni-Rached, Michel

2016-11-01

Frozen waves (FWs) are a class of diffraction- and attenuation-resistant beams whose intensity pattern along the direction of propagation can be chosen arbitrarily, thus making them relevant for engineering the spatial configuration of optical fields. To date, analyses of such beams have been done essentially for the scalar case, with the vectorial nature of the electromagnetic fields often neglected. Although it is expected that the field components keep the fundamental properties of the scalar FWs, a deeper understanding of their electromagnetic counterparts is mandatory in order to exploit their different possible polarization states. The purpose of this paper is to study the properties of electromagnetic FWs with radial, azimuthal, linear, circular, and elliptical polarizations under paraxial and nonparaxial regimes in nonabsorbing media. An intensity pattern is chosen for a scalar FW, and the vectorial solutions are built after it via the use of Maxwell's equations. The results show that the field components and the longitudinal component of the time-averaged Poynting vector closely follow the pattern chosen even under highly nonparaxial conditions, showing the robustness of the FW structure to parameters variations.

Vectorial Command of Induction Motor Pumping System Supplied by a Photovoltaic Generator

NASA Astrophysics Data System (ADS)

Makhlouf, Messaoud; Messai, Feyrouz; Benalla, Hocine

2011-01-01

With the continuous decrease of the cost of solar cells, there is an increasing interest and needs in photovoltaic (PV) system applications following standard of living improvements. Water pumping system powered by solar-cell generators are one of the most important applications. The fluctuation of solar energy on one hand, and the necessity to optimise available solar energy on the other, it is useful to develop new efficient and flexible modes to control motors that entrain the pump. A vectorial control of an asynchronous motor fed by a photovoltaic system is proposed. This paper investigates a photovoltaic-electro mechanic chain, composed of a PV generator, DC-AC converter, a vector controlled induction motor and centrifugal pump. The PV generator is forced to operate at its maximum power point by using an appropriate search algorithm integrated in the vector control. The optimization is realized without need to adding a DC-DC converter to the chain. The motor supply is also ensured in all insolation conditions. Simulation results show the effectiveness and feasibility of such an approach.

Wall shear stress fixed points in cardiovascular fluid mechanics.

PubMed

Arzani, Amirhossein; Shadden, Shawn C

2018-05-17

Complex blood flow in large arteries creates rich wall shear stress (WSS) vectorial features. WSS acts as a link between blood flow dynamics and the biology of various cardiovascular diseases. WSS has been of great interest in a wide range of studies and has been the most popular measure to correlate blood flow to cardiovascular disease. Recent studies have emphasized different vectorial features of WSS. However, fixed points in the WSS vector field have not received much attention. A WSS fixed point is a point on the vessel wall where the WSS vector vanishes. In this article, WSS fixed points are classified and the aspects by which they could influence cardiovascular disease are reviewed. First, the connection between WSS fixed points and the flow topology away from the vessel wall is discussed. Second, the potential role of time-averaged WSS fixed points in biochemical mass transport is demonstrated using the recent concept of Lagrangian WSS structures. Finally, simple measures are proposed to quantify the exposure of the endothelial cells to WSS fixed points. Examples from various arterial flow applications are demonstrated. Copyright © 2018 Elsevier Ltd. All rights reserved.

Gravity dependency of the gramicidin A channel conductivity. A model for gravity perception on the cellular level.

PubMed

Schatz, A; Linke-Hommes, A; Neubert, J

1996-01-01

Theoretical investigations involving the membrane-solution interface have revealed that the density of the solution varies appreciably within interfacial layers adjacent to charged membrane surfaces. The hypothesis that gravity interacts with this configuration and modifies transport rates across horizontal and vertical membranes differently was supported by initial experiments with gramicidin A channels in phosphatidylserine (PS) membranes in 0.1 M KCl. Channel conductivity was found to be about 1.6 times higher in horizontal membranes than in vertical membranes. Here we present the results of further experiments with gramicidin A channels (incorporated into charged PS- and uncharged phosphatidylcholine (PC) membranes in KCl- and CsCl-solutions) to demonstrate that the hypothesis is more generally applicable. Again, channel conductivity was found to be higher in horizontal PS membranes by a factor of between 1.20 and 1.75 in 0.1 M CsCl. No difference in channel conductivity was found for uncharged PC membranes in 0.1 M KCl and in 0.1 M CsCl. However, for PC membranes in 0.05 M KCl the channel conductivity was significantly higher in horizontal membranes by a factor of between 1.07 and 1.14. These results are consistent with the results of our model calculations of layer density and extension, which showed that the layer formation is enhanced by increasing membrane surface charge and decreasing electrolyte ion concentration. The mechanism of gravity interaction with membrane transport processes via interface reactions might be utilized by biological systems for orientational behaviour in the gravity field, which has been observed even for cellular systems.

Structure and Dynamics of the Membrane-Bound Cytochrome P450 2C9

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

Cojocaru, Vlad; Balali-Mood, Kia; Sansom, Mark S.

The microsomal, membrane-bound, human cytochrome P450 (CYP) 2C9 is a liver-specific monooxygenase essential for drug metabolism. CYPs require electron transfer from the membrane-bound CYP reductase (CPR) for catalysis. The structural details and functional relevance of the CYP-membrane interaction are not understood. From multiple coarse grained molecular simulations started with arbitrary configurations of protein-membrane complexes, we found two predominant orientations of CYP2C9 in the membrane, both consistent with experiments and conserved in atomic-resolution simulations. The dynamics of membrane-bound and soluble CYP2C9 revealed correlations between opening and closing of different tunnels from the enzyme’s buried active site. The membrane facilitated the openingmore » of a tunnel leading into it by stabilizing the open state of an internal aromatic gate. Other tunnels opened selectively in the simulations of product-bound CYP2C9. We propose that the membrane promotes binding of liposoluble substrates by stabilizing protein conformations with an open access tunnel and provide evidence for selective substrate access and product release routes in mammalian CYPs. The models derived here are suitable for extension to incorporate other CYPs for oligomerization studies or the CYP reductase for studies of the electron transfer mechanism, whereas the modeling procedure is generally applicable to study proteins anchored in the bilayer by a single transmembrane helix.« less

Effect of phloretin on the permeability of thin lipid membranes

PubMed Central

1976-01-01

Phloretin dramatically increases cation conductances and decreases anion conductances of membranes treated with ion carriers (nonactin, valinomycin, carbonyl-cyanide-m-chlorophenylhydrazone [CCCP], and Hg(C6F5)2) or lipophilic ions (tetraphenylarsonium [tphAs+] and tetraphenylborate [TPhB-]). For example, on phosphatidylethanolamine membranes, 10(-4) M phloretin increases K+ -nonactin and TPhAs+ conductances and decreases CCCP- and TPhB- conductances 10(3)-fold; on lecithin: cholesterol membranes, it increases K+-nonactin conductance 10(5)-fold and decreases CCCP- conductance 10(3)-fold. Similar effects are obtained with p- and m-nitrophenol at 10(-2) M. These effects are produced by the un-ionized form of phloretin and the nitrophenols. We believe that phloretin, which possesses a large dipole moment, adsorbs and orients at the membrane surface to introduce a dipole potential of opposite polarity to the preexisting positive one, thus increasing the partition coefficient of cations into the membrane interior and decreasing the partition coefficient of anions. (Phloretin may also increase the fluidity of cholesterol-containing membranes; this is manifested by its two- to three-fold increase in nonelectrolyte permeability and its asymmetrical effect on cation and anion conductances in cholesterol-containing membranes.) It is possible that pholoretin's inhibition of chloride, urea, and glucose transport in biological membranes results from the effects of these intense intrafacial dipole fields on the translocator(s) of these molecules. PMID:946975

Differential polarization laser scanning microscopy: biological applications

NASA Astrophysics Data System (ADS)

Steinbach, G.; Besson, F.; Pomozi, I.; Garab, G.

2005-09-01

With the aid of a differential polarization (DP) apparatus, developed in our laboratory and attached to our laser scanning confocal microscope, we can measure the magnitude and spatial distribution of 8 different DP quantities: linear and circular dichroism (LD&CD), linear and circular anisotropy of the emission (R and CPL, confocal), fluorescence detected dichroisms (FDLD&FDCD, confocal), linear birefringence (LB), and the degree of polarization of fluorescence emission (P, confocal). The attachment uses high frequency modulation and subsequent demodulation, via lock-in amplifier, of the detected intensity values, and records and displays pixel-by-pixel the measured DP quantity. These microscopic DP data carry important physical information on the molecular architecture of anisotropically organized samples. Microscopic DP measurements are thought to be of particular importance in biology. In most biological samples anisotropy is difficult to determine with conventional, macroscopic DP measurements and microscopic variations are of special significance. In this paper, we describe the method of LB imaging. Using magnetically oriented isolated chloroplasts trapped in polyacrylamide gel, we demonstrate that LB can be determined with high sensitivity and good spatial resolution. Granal thylakoid membranes in edge-aligned orientation exhibited strong LB, with large variations in its sign and magnitude. In face-aligned position LB was considerably weaker, and tended to vanish when averaged for the whole image. The strong local variations are attributed to the inherent heterogeneity of the membranes, i.e. to their internal differentiation into multilamellar, stacked membranes (grana), and single thylakoids (stroma membranes). Further details and applications of our DP-LSM will be published elsewhere.

Integrated Analysis of Environment, Cattle and Human Serological Data: Risks and Mechanisms of Transmission of Rift Valley Fever in Madagascar

PubMed Central

Olive, Marie-Marie; Chevalier, Véronique; Grosbois, Vladimir; Tran, Annelise; Andriamandimby, Soa-Fy; Durand, Benoit; Ravalohery, Jean-Pierre; Andriamamonjy, Seta; Rakotomanana, Fanjasoa; Rogier, Christophe; Heraud, Jean-Michel

2016-01-01

Background Rift Valley fever (RVF) is a vector-borne disease affecting ruminants and humans. Madagascar was heavily affected by RVF in 2008–2009, with evidence of a large and heterogeneous spread of the disease. The identification of at-risk environments is essential to optimize the available resources by targeting RVF surveillance in Madagascar. Herein, the objectives of our study were: (i) to identify the environmental factors and areas favorable to RVF transmission to both cattle and human and (ii) to identify human behaviors favoring human infections in Malagasy contexts. Methodology/Principal Findings First, we characterized the environments of Malagasy communes using a Multiple Factor Analysis (MFA). Then, we analyzed cattle and human serological data collected at national level using Generalized Linear Mixed Models, with the individual serological status (cattle or human) as the response, and MFA factors, as well as other potential risk factors (cattle density, human behavior) as explanatory variables. Cattle and human seroprevalence rates were positively associated to humid environments (p<0.001). Areas with high cattle density were at risk (p<0.01; OR = 2.6). Furthermore, our analysis showed that frequent contact with raw milk contributed to explain human infection (OR = 1.6). Finally, our study highlighted the eastern-coast, western and north-western parts as high-risk areas for RVF transmission in cattle. Conclusions/Significance Our integrated approach analyzing environmental, cattle and human datasets allow us to bring new insight on RVF transmission patterns in Madagascar. The association between cattle seroprevalence, humid environments and high cattle density suggests that concomitant vectorial and direct transmissions are critical to maintain RVF enzootic transmission. Additionally, in the at-risk humid environment of the western, north-western and the eastern-coast areas, suitable to Culex and Anopheles mosquitoes, vectorial transmission probably occurs in both cattle and human. The relative contribution of vectorial or direct transmissions could be further assessed by mathematic modelling. PMID:27415438

Integrated Analysis of Environment, Cattle and Human Serological Data: Risks and Mechanisms of Transmission of Rift Valley Fever in Madagascar.

PubMed

Olive, Marie-Marie; Chevalier, Véronique; Grosbois, Vladimir; Tran, Annelise; Andriamandimby, Soa-Fy; Durand, Benoit; Ravalohery, Jean-Pierre; Andriamamonjy, Seta; Rakotomanana, Fanjasoa; Rogier, Christophe; Heraud, Jean-Michel

2016-07-01

Rift Valley fever (RVF) is a vector-borne disease affecting ruminants and humans. Madagascar was heavily affected by RVF in 2008-2009, with evidence of a large and heterogeneous spread of the disease. The identification of at-risk environments is essential to optimize the available resources by targeting RVF surveillance in Madagascar. Herein, the objectives of our study were: (i) to identify the environmental factors and areas favorable to RVF transmission to both cattle and human and (ii) to identify human behaviors favoring human infections in Malagasy contexts. First, we characterized the environments of Malagasy communes using a Multiple Factor Analysis (MFA). Then, we analyzed cattle and human serological data collected at national level using Generalized Linear Mixed Models, with the individual serological status (cattle or human) as the response, and MFA factors, as well as other potential risk factors (cattle density, human behavior) as explanatory variables. Cattle and human seroprevalence rates were positively associated to humid environments (p<0.001). Areas with high cattle density were at risk (p<0.01; OR = 2.6). Furthermore, our analysis showed that frequent contact with raw milk contributed to explain human infection (OR = 1.6). Finally, our study highlighted the eastern-coast, western and north-western parts as high-risk areas for RVF transmission in cattle. Our integrated approach analyzing environmental, cattle and human datasets allow us to bring new insight on RVF transmission patterns in Madagascar. The association between cattle seroprevalence, humid environments and high cattle density suggests that concomitant vectorial and direct transmissions are critical to maintain RVF enzootic transmission. Additionally, in the at-risk humid environment of the western, north-western and the eastern-coast areas, suitable to Culex and Anopheles mosquitoes, vectorial transmission probably occurs in both cattle and human. The relative contribution of vectorial or direct transmissions could be further assessed by mathematic modelling.

[Effect of the number of health meals before an infectious meal on the vectorial competence of Glossina morsitans morsitans infected by Trypanosoma congolense IL 1180].

PubMed

Kazadi, J M; Kageruka, P; Losson, B

1999-01-01

The purpose of this work was to assess the influence of several healthy meals (0, 1 and 2) prior to the infectious one on the vectorial competence of Glossina morsitans morsitans (Mall). The teneral flies (< 32 h old) of this line were divided into three groups. The tsetse flies of group A received no meal. The ones of group B received one healthy meal on day 1, whereas those from group C were given two consecutive healthy meals on days 1 and 2. All the flies were experimentally infected with Trypanosoma congolense IL 1180 when the maximum age reached 32 h for flies with no meal, 56 h for those with one healthy meal and 80 h for those who received two healthy meals. When both sexes were considered, the meso-procyclic and metacyclic indexes as well as the vectorial competence (VC) of the flies receiving no meal were 0.99 +/- 0.01, 0.96 +/- 0.02 and 0.95 +/- 0.03. Considering the flies which were fed one healthy meal, the respective values were 0.42 +/- 0.13, 0.50 +/- 0.01 and 0.21 +/- 0.06, whereas the values for the flies receiving two healthy meals were 0.45 +/- 0.11, 0.29 +/- 0.19 and 0.13 +/- 0.05. The meso-procyclic and metacyclic indexes as well as the VC in both sexes were more important in the flies which received no meal than those fed with one or two healthy meals. The meso-procyclic and metacyclic indexes and VC did not show any significant differences between the flies fed one or two healthy meals, whereas the metacyclic index of male flies which received one healthy meal was significantly higher than those fed two healthy meals. These results indicate that the number of non-infected (healthy) meals prior to an infected meal reduces the interaction between G. m. morsitans infected and T. congolense.

Are the London Declaration's 2020 goals sufficient to control Chagas disease?: Modeling scenarios for the Yucatan Peninsula.

PubMed

Lee, Bruce Y; Bartsch, Sarah M; Skrip, Laura; Hertenstein, Daniel L; Avelis, Cameron M; Ndeffo-Mbah, Martial; Tilchin, Carla; Dumonteil, Eric O; Galvani, Alison

2018-03-01

The 2020 Sustainable Development goals call for 100% certified interruption or control of the three main forms of Chagas disease transmission in Latin America. However, how much will achieving these goals to varying degrees control Chagas disease; what is the potential impact of missing these goals and if they are achieved, what may be left? We developed a compartmental simulation model that represents the triatomine, human host, and non-human host populations and vector-borne, congenital, and transfusional T. cruzi transmission between them in the domestic and peridomestic settings to evaluate the impact of limiting transmission in a 2,000 person virtual village in Yucatan, Mexico. Interruption of domestic vectorial transmission had the largest impact on T. cruzi transmission and prevalence in all populations. Most of the gains were achieved within the first few years. Controlling vectorial transmission resulted in a 46.1-83.0% relative reduction in the number of new acute Chagas cases for a 50-100% interruption in domestic vector-host contact. Only controlling congenital transmission led to a 2.4-8.1% (30-100% interruption) relative reduction in the total number of new acute cases and reducing only transfusional transmission led to a 0.1-0.3% (30-100% reduction). Stopping all three forms of transmission resulted in 0.5 total transmission events over five years (compared to 5.0 with no interruption); interrupting all forms by 30% resulted in 3.4 events over five years per 2,000 persons. While reducing domestic vectorial, congenital, and transfusional transmission can successfully reduce transmission to humans (up to 82% in one year), achieving the 2020 goals would still result in 0.5 new acute cases per 2,000 over five years. Even if the goals are missed, major gains can be achieved within the first few years. Interrupting transmission should be combined with other efforts such as a vaccine or improved access to care, especially for the population of already infected individuals.

Fiber-chip edge coupler with large mode size for silicon photonic wire waveguides.

PubMed

Papes, Martin; Cheben, Pavel; Benedikovic, Daniel; Schmid, Jens H; Pond, James; Halir, Robert; Ortega-Moñux, Alejandro; Wangüemert-Pérez, Gonzalo; Ye, Winnie N; Xu, Dan-Xia; Janz, Siegfried; Dado, Milan; Vašinek, Vladimír

2016-03-07

Fiber-chip edge couplers are extensively used in integrated optics for coupling of light between planar waveguide circuits and optical fibers. In this work, we report on a new fiber-chip edge coupler concept with large mode size for silicon photonic wire waveguides. The coupler allows direct coupling with conventional cleaved optical fibers with large mode size while circumventing the need for lensed fibers. The coupler is designed for 220 nm silicon-on-insulator (SOI) platform. It exhibits an overall coupling efficiency exceeding 90%, as independently confirmed by 3D Finite-Difference Time-Domain (FDTD) and fully vectorial 3D Eigenmode Expansion (EME) calculations. We present two specific coupler designs, namely for a high numerical aperture single mode optical fiber with 6 µm mode field diameter (MFD) and a standard SMF-28 fiber with 10.4 µm MFD. An important advantage of our coupler concept is the ability to expand the mode at the chip edge without leading to high substrate leakage losses through buried oxide (BOX), which in our design is set to 3 µm. This remarkable feature is achieved by implementing in the SiO 2 upper cladding thin high-index Si 3 N 4 layers. The Si 3 N 4 layers increase the effective refractive index of the upper cladding near the facet. The index is controlled along the taper by subwavelength refractive index engineering to facilitate adiabatic mode transformation to the silicon wire waveguide while the Si-wire waveguide is inversely tapered along the coupler. The mode overlap optimization at the chip facet is carried out with a full vectorial mode solver. The mode transformation along the coupler is studied using 3D-FDTD simulations and with fully-vectorial 3D-EME calculations. The couplers are optimized for operating with transverse electric (TE) polarization and the operating wavelength is centered at 1.55 µm.

A Novel HMM Distributed Classifier for the Detection of Gait Phases by Means of a Wearable Inertial Sensor Network

PubMed Central

Taborri, Juri; Rossi, Stefano; Palermo, Eduardo; Patanè, Fabrizio; Cappa, Paolo

2014-01-01

In this work, we decided to apply a hierarchical weighted decision, proposed and used in other research fields, for the recognition of gait phases. The developed and validated novel distributed classifier is based on hierarchical weighted decision from outputs of scalar Hidden Markov Models (HMM) applied to angular velocities of foot, shank, and thigh. The angular velocities of ten healthy subjects were acquired via three uni-axial gyroscopes embedded in inertial measurement units (IMUs) during one walking task, repeated three times, on a treadmill. After validating the novel distributed classifier and scalar and vectorial classifiers-already proposed in the literature, with a cross-validation, classifiers were compared for sensitivity, specificity, and computational load for all combinations of the three targeted anatomical segments. Moreover, the performance of the novel distributed classifier in the estimation of gait variability in terms of mean time and coefficient of variation was evaluated. The highest values of specificity and sensitivity (>0.98) for the three classifiers examined here were obtained when the angular velocity of the foot was processed. Distributed and vectorial classifiers reached acceptable values (>0.95) when the angular velocity of shank and thigh were analyzed. Distributed and scalar classifiers showed values of computational load about 100 times lower than the one obtained with the vectorial classifier. In addition, distributed classifiers showed an excellent reliability for the evaluation of mean time and a good/excellent reliability for the coefficient of variation. In conclusion, due to the better performance and the small value of computational load, the here proposed novel distributed classifier can be implemented in the real-time application of gait phases recognition, such as to evaluate gait variability in patients or to control active orthoses for the recovery of mobility of lower limb joints. PMID:25184488

Molecular order and T1-relaxation, cross-relaxation in nitroxide spin labels

NASA Astrophysics Data System (ADS)

Marsh, Derek

2018-05-01

Interpretation of saturation-recovery EPR experiments on nitroxide spin labels whose angular rotation is restricted by the orienting potential of the environment (e.g., membranes) currently concentrates on the influence of rotational rates and not of molecular order. Here, I consider the dependence on molecular ordering of contributions to the rates of electron spin-lattice relaxation and cross relaxation from modulation of N-hyperfine and Zeeman anisotropies. These are determined by the averages and , where θ is the angle between the nitroxide z-axis and the static magnetic field, which in turn depends on the angles that these two directions make with the director of uniaxial ordering. For saturation-recovery EPR at 9 GHz, the recovery rate constant is predicted to decrease with increasing order for the magnetic field oriented parallel to the director, and to increase slightly for the perpendicular field orientation. The latter situation corresponds to the usual experimental protocol and is consistent with the dependence on chain-labelling position in lipid bilayer membranes. An altered dependence on order parameter is predicted for saturation-recovery EPR at high field (94 GHz) that is not entirely consistent with observation. Comparisons with experiment are complicated by contributions from slow-motional components, and an unexplained background recovery rate that most probably is independent of order parameter. In general, this analysis supports the interpretation that recovery rates are determined principally by rotational diffusion rates, but experiments at other spectral positions/field orientations could increase the sensitivity to order parameter.

Ligand structure and mechanical properties of single-nanoparticle-thick membranes.

PubMed

Salerno, K Michael; Bolintineanu, Dan S; Lane, J Matthew D; Grest, Gary S

2015-06-01

The high mechanical stiffness of single-nanoparticle-thick membranes is believed to result from the local structure of ligand coatings that mediate interactions between nanoparticles. These ligand structures are not directly observable experimentally. We use molecular dynamics simulations to observe variations in ligand structure and simultaneously measure variations in membrane mechanical properties. We have shown previously that ligand end group has a large impact on ligand structure and membrane mechanical properties. Here we introduce and apply quantitative molecular structure measures to these membranes and extend analysis to multiple nanoparticle core sizes and ligand lengths. Simulations of nanoparticle membranes with a nanoparticle core diameter of 4 or 6 nm, a ligand length of 11 or 17 methylenes, and either carboxyl (COOH) or methyl (CH(3)) ligand end groups are presented. In carboxyl-terminated ligand systems, structure and interactions are dominated by an end-to-end orientation of ligands. In methyl-terminated ligand systems large ordered ligand structures form, but nanoparticle interactions are dominated by disordered, partially interdigitated ligands. Core size and ligand length also affect both ligand arrangement within the membrane and the membrane's macroscopic mechanical response, but are secondary to the role of the ligand end group. Moreover, the particular end group (COOH or CH(3)) alters the nature of how ligand length, in turn, affects the membrane properties. The effect of core size does not depend on the ligand end group, with larger cores always leading to stiffer membranes. Asymmetry in the stress and ligand density is observed in membranes during preparation at a water-vapor interface, with the stress asymmetry persisting in all membranes after drying.

The transport along membrane nanotubes driven by the spontaneous curvature of membrane components.

PubMed

Kabaso, Doron; Bobrovska, Nataliya; Góźdź, Wojciech; Gongadze, Ekaterina; Kralj-Iglič, Veronika; Zorec, Robert; Iglič, Aleš

2012-10-01

Intercellular membrane nanotubes (ICNs) serve as a very specific transport system between neighboring cells. The underlying mechanisms responsible for the transport of membrane components and vesicular dilations along the ICNs are not clearly understood. The present study investigated the spatial distribution of anisotropic membrane components of tubular shapes and isotropic membrane components of spherical shapes. Experimental results revealed the preferential distribution of CTB (cholera toxin B)-GM1 complexes mainly on the spherical cell membrane, and cholesterol-sphingomyelin at the membrane leading edge and ICNs. In agreement with previous studies, we here propose that the spatial distribution of CTB-GM1 complexes and cholesterol-sphingomyelin rafts were due to their isotropic and anisotropic shapes, respectively. To elucidate the relationship between a membrane component shape and its spatial distribution, a two-component computational model was constructed. The minimization of the membrane bending (free) energy revealed the enrichment of the anisotropic component along the ICN and the isotropic component in the parent cell membrane, which was due to the curvature mismatch between the ICN curvature and the spontaneous curvature of the isotropic component. The equations of motion, derived from the differentiation of the membrane free energy, revealed a curvature-dependent flux of the isotropic component and a curvature-dependent force exerted on a vesicular dilation along the ICN. Finally, the effects of possible changes in the orientational ordering of the anisotropic component attendant to the transport of the vesicular dilation were discussed with connection to the propagation of electrical and chemical signals. Copyright © 2012 Elsevier B.V. All rights reserved.

Assembly of purple membranes on polyelectrolyte films.

PubMed

Saab, Marie-belle; Estephan, Elias; Cloitre, Thierry; Legros, René; Cuisinier, Frédéric J G; Zimányi, László; Gergely, Csilla

2009-05-05

The membrane protein bacteriorhodopsin in its native membrane bound form (purple membrane) was adsorbed and incorporated into polyelectrolyte multilayered films, and adsorption was in situ monitored by optical waveguide light-mode spectroscopy. The formation of a single layer or a double layer of purple membranes was observed when adsorbed on negatively or positively charged surfaces, respectively. The purple membrane patches adsorbed on the polyelectrolyte multilayers were also evidenced by atomic force microscopy images. The driving forces of the adsorption process were evaluated by varying the ionic strength of the solution as well as the purple membrane concentration. At high purple membrane concentration, interpenetrating polyelectrolyte loops might provide new binding sites for the adsorption of a second layer of purple membranes, whereas at lower concentrations only a single layer is formed. Negative surfaces do not promote a second protein layer adsorption. Driving forces other than just electrostatic ones, such as hydrophobic forces, should play a role in the polyelectrolyte/purple membrane layering. The subtle interplay of all these factors determines the formation of the polyelectrolyte/purple membrane matrix with a presumably high degree of orientation for the incorporated purple membranes, with their cytoplasmic, or extracellular side toward the bulk on negatively or positively charged polyelectrolyte, respectively. The structural stability of bacteriorhodopsin during adsorption onto the surface and incorporation into the polyelectrolyte multilayers was investigated by Fourier transform infrared spectroscopy in attenuated total reflection mode. Adsorption and incorporation of purple membranes within polyelectrolyte multilayers does not disturb the conformational majority of membrane-embedded alpha-helix structures of the protein, but may slightly alter the structure of the extramembraneous segments or their interaction with the environment. This high stability is different from the lower stability of the predominantly beta-sheet structures of numerous globular proteins when adsorbed onto surfaces.

Two-dimensional protein crystals (S-layers): fundamentals and applications.

PubMed

Sleytr, U B; Sára, M; Messner, P; Pum, D

1994-10-01

Two-dimensional crystalline surface layers (S-layers) composed of protein or glycoprotein subunits are one of the most commonly observed prokaryotic cell envelope structures. Isolated S-layer subunits are endowed with the ability to assemble into monomolecular arrays in suspension, on surfaces or interfaces by an entropy-driven process. S-layer lattices are isoporous structures with functional groups located on the surface in an identical position and orientation. These characteristic features have already led to applications of S-layers as (1) ultrafiltration membranes with well-defined molecular weight cut-offs and excellent antifouling characteristics, (2) immobilization matrices for functional molecules as required for affinity and enzyme membranes, affinity microcarriers and biosensors, (3) conjugate vaccines, (4) carriers for Langmuir-Blodgett films and reconstituted biological membranes, and (5) patterning elements in molecular nanotechnology.

Morphological features (defects) in fuel cell membrane electrode assemblies

NASA Astrophysics Data System (ADS)

Kundu, S.; Fowler, M. W.; Simon, L. C.; Grot, S.

Reliability and durability issues in fuel cells are becoming more important as the technology and the industry matures. Although research in this area has increased, systematic failure analysis, such as a failure modes and effects analysis (FMEA), are very limited in the literature. This paper presents a categorization scheme of causes, modes, and effects related to fuel cell degradation and failure, with particular focus on the role of component quality, that can be used in FMEAs for polymer electrolyte membrane (PEM) fuel cells. The work also identifies component defects imparted on catalyst-coated membranes (CCM) by manufacturing and proposes mechanisms by which they can influence overall degradation and reliability. Six major defects have been identified on fresh CCM materials, i.e., cracks, orientation, delamination, electrolyte clusters, platinum clusters, and thickness variations.

Miniaturized ceramic platform for metal oxide gas sensors array

NASA Astrophysics Data System (ADS)

Samotaev, N. N.

2016-10-01

In work is developing an ultra-fast, low cost and technology flexible process for production array of ceramic MEMS microhotplates for using in semiconductor gas sensors orientated to small series applications, where is sufficient to produce 10-100 samples with a different layout of heaters and membrane per day.

Membrane transporters studied by EPR spectroscopy: structure determination and elucidation of functional dynamics.

PubMed

Mullen, Anna; Hall, Jenny; Diegel, Janika; Hassan, Isa; Fey, Adam; MacMillan, Fraser

2016-06-15

During their mechanistic cycles membrane transporters often undergo extensive conformational changes, sampling a range of orientations, in order to complete their function. Such membrane transporters present somewhat of a challenge to conventional structural studies; indeed, crystallization of membrane-associated proteins sometimes require conditions that vary vastly from their native environments. Moreover, this technique currently only allows for visualization of single selected conformations during any one experiment. EPR spectroscopy is a magnetic resonance technique that offers a unique opportunity to study structural, environmental and dynamic properties of such proteins in their native membrane environments, as well as readily sampling their substrate-binding-induced dynamic conformational changes especially through complementary computational analyses. Here we present a review of recent studies that utilize a variety of EPR techniques in order to investigate both the structure and dynamics of a range of membrane transporters and associated proteins, focusing on both primary (ABC-type transporters) and secondary active transporters which were key interest areas of the late Professor Stephen Baldwin to whom this review is dedicated. © 2016 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Polarizing intestinal epithelial cells electrically through Ror2

PubMed Central

Cao, Lin; McCaig, Colin D.; Scott, Roderick H.; Zhao, Siwei; Milne, Gillian; Clevers, Hans; Zhao, Min; Pu, Jin

2014-01-01

ABSTRACT The apicobasal polarity of enterocytes determines where the brush border membrane (apical membrane) will form, but how this apical membrane faces the lumen is not well understood. The electrical signal across the epithelium could serve as a coordinating cue, orienting and polarizing enterocytes. Here, we show that applying a physiological electric field to intestinal epithelial cells, to mimic the natural electric field created by the transepithelial potential difference, polarized phosphorylation of the actin-binding protein ezrin, increased expression of intestinal alkaline phosphatase (ALPI, a differentiation marker) and remodeled the actin cytoskeleton selectively on the cathode side. In addition, an applied electric field also activated ERK1/2 and LKB1 (also known as STK11), key molecules in apical membrane formation. Disruption of the tyrosine protein kinase transmembrane receptor Ror2 suppressed activation of ERK1/2 and LKB1 significantly, and subsequently inhibited apical membrane formation in enterocytes. Our findings indicate that the endogenous electric field created by the transepithelial potential difference might act as an essential coordinating signal for apical membrane formation at a tissue level, through activation of LKB1 mediated by Ror2–ERK signaling. PMID:24928904

Structurally detailed coarse-grained model for Sec-facilitated co-translational protein translocation and membrane integration

PubMed Central

Miller, Thomas F.

2017-01-01

We present a coarse-grained simulation model that is capable of simulating the minute-timescale dynamics of protein translocation and membrane integration via the Sec translocon, while retaining sufficient chemical and structural detail to capture many of the sequence-specific interactions that drive these processes. The model includes accurate geometric representations of the ribosome and Sec translocon, obtained directly from experimental structures, and interactions parameterized from nearly 200 μs of residue-based coarse-grained molecular dynamics simulations. A protocol for mapping amino-acid sequences to coarse-grained beads enables the direct simulation of trajectories for the co-translational insertion of arbitrary polypeptide sequences into the Sec translocon. The model reproduces experimentally observed features of membrane protein integration, including the efficiency with which polypeptide domains integrate into the membrane, the variation in integration efficiency upon single amino-acid mutations, and the orientation of transmembrane domains. The central advantage of the model is that it connects sequence-level protein features to biological observables and timescales, enabling direct simulation for the mechanistic analysis of co-translational integration and for the engineering of membrane proteins with enhanced membrane integration efficiency. PMID:28328943

Transport of N-acetyl-S-(1,2-dichlorovinyl)-L-cysteine, a metabolite of trichloroethylene, by mouse multidrug resistance associated protein 2 (Mrp2)

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

Tsirulnikov, Kirill; Abuladze, Natalia; Koag, Myong-Chul

2010-04-15

N-acetyl-S-(1,2-dichlorovinyl)-L-cysteine (Ac-DCVC) and S-(1,2-dichlorovinyl)-L-cysteine (DCVC) are the glutathione conjugation pathway metabolites of a common industrial contaminant and potent nephrotoxicant trichloroethylene (TCE). Ac-DCVC and DCVC are accumulated in the renal proximal tubule where they may be secreted into the urine by an unknown apical transporter(s). In this study, we explored the hypothesis that the apical transport of Ac-DCVC and/or DCVC may be mediated by the multidrug resistance associated protein 2 (Mrp2, ABCC2), which is known to mediate proximal tubular apical ATP-dependent transport of glutathione and numerous xenobiotics and endogenous substances conjugated with glutathione. Transport experiments using membrane vesicles prepared from mousemore » proximal tubule derived cells expressing mouse Mrp2 utilizing ATPase assay and direct measurements of Ac-DCVC/DCVC using liquid chromatography/tandem mass-spectrometry (LC/MS/MS) demonstrated that mouse Mrp2 mediates ATP-dependent transport of Ac-DCVC. Expression of mouse Mrp2 antisense mRNA significantly inhibited the vectorial basolateral to apical transport of Ac-DCVC but not DCVC in mouse proximal tubule derived cells endogenously expressing mouse Mrp2. The results suggest that Mrp2 may be involved in the renal secretion of Ac-DCVC.« less

Self-Repair and Patterning of 2D Membrane-Like Peptoid Materials

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

Jiao, Fang; Chen, Yulin; Jin, Haibao

2016-08-31

Two-dimensional materials are of increasing interest for use in filtration, sensing, nanoelectronics, and biomedical devices. Peptoids are a class of biomimetic sequence-defined polymers for which certain amphiphillic sequences self-assemble into 2D crystalline materials with properties that mimic those of cell membranes. Using in situ AFM to both dissect these membrane-like materials and image their subsequent behavior, we explore their ability to self-repair on a range of solid substrates. We show that, in a suitable range of pH, self-repair occurs on both negatively and positively charged substrates and can even occur in the absence of an underlying surface. Following dissection ofmore » pre-assembled peptoid membranes and upon introduction of a peptoid monomer solution, peptoids repair the damage by assembling at the newly created edges. The speed of the advancing edge depends on the edge orientation, reflecting the two-fold symmetry of the underlying peptoid lattice. Moreover, because the membranes are stabilized by hydrophobic interactions, if the solution contains peptoids possessing an identical sequence in the hydrophobic block but a distinct hydrophilic block, filling of the defects creates membranes that are patterned at the nanoscale. Consequently, we can utilize this ability to create nm-sized patterns of distinct functional groups within a single coherent membrane.« less

The role of eggshell and underlying vitelline membrane for normal pattern formation in the early C. elegans embryo.

PubMed

Schierenberg, Einhard; Junkersdorf, Bernd

1992-12-01

The embryo of the nematode Caenorhabditis elegans is surrounded by an inconspicuous inner vitelline membrane and a prominent outer chitinous eggshell proper. We demonstrate that the complete removal of the chitinous eggshell does not interfere with successful development to yield a normal worm. The same result can be obtained when the vitelline membrane is penetrated with laser microbeam irradiation of only the eggshell proper, gently enough to permit its resealing after a while. However, when large holes are made into the eggshell the concomitantly penetrated vitelline membrane does not reseal. While early development is quite normal under these conditions, gastrulation is defective in that gut precursor cells do not migrate in properly, eventually leading to embryonic arrest. This suggests a crucial role for pattern formation of the "micro-environment" around the embryo preserved by the intact vitelline membrane. Removing both eggshell and vitelline membrane results in a string-like arrangement of founder cells and subsequent grossly abnormal cell patterns. Our experiments support the idea that the prominent eggshell proper just functions as a mechanical protection while the thin vitelline membrane directly or indirectly serves as a necessary control element affecting the positions of cells which to begin with are determined by the orientation of the cleavage spindle.

Process-Oriented Review of Bacterial Quorum Quenching for Membrane Biofouling Mitigation in Membrane Bioreactors (MBRs)

PubMed Central

Bouayed, Naila; Dietrich, Nicolas; Lafforgue, Christine; Lee, Chung-Hak; Guigui, Christelle

2016-01-01

Quorum Quenching (QQ) has been developed over the last few years to overcome practical issues related to membrane biofouling, which is currently the major difficulty thwarting the extensive development of membrane bioreactors (MBRs). QQ is the disruption of Quorum Sensing (QS), cell-to-cell communication enabling the bacteria to harmonize their behavior. The production of biofilm, which is recognized as a major part of the biocake formed on a membrane surface, and which leads to biofouling, has been found to be one of the bacterial behaviors controlled by QS. Since the enzymatic disruption of QS was reported to be efficient as a membrane biofouling mitigation technique in MBRs, the application of QQ to lab-scale MBRs has been the subject of much research using different approaches under different operating conditions. This paper gives an overview of the effectiveness of QQ in mitigating membrane biofouling in MBRs. It is based on the results of previous studies, using two microbial strains, Rhodococcus sp. BH4 and Pseudomonas sp. 1A1. The effect of bacterial QQ on the physical phenomena of the MBR process is analyzed, adopting an original multi-scale approach. Finally, the potential influence of the MBR operating conditions on QQ effectiveness is discussed. PMID:27983578