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Sample records for large-aperture membrane active

  1. Large-Aperture Membrane Active Phased-Array Antennas

    NASA Technical Reports Server (NTRS)

    Karasik, Boris; McGrath, William; Leduc, Henry

    2009-01-01

    Large-aperture phased-array microwave antennas supported by membranes are being developed for use in spaceborne interferometric synthetic aperture radar systems. There may also be terrestrial uses for such antennas supported on stationary membranes, large balloons, and blimps. These antennas are expected to have areal mass densities of about 2 kg/sq m, satisfying a need for lightweight alternatives to conventional rigid phased-array antennas, which have typical areal mass densities between 8 and 15 kg/sq m. The differences in areal mass densities translate to substantial differences in total mass in contemplated applications involving aperture areas as large as 400 sq m. A membrane phased-array antenna includes patch antenna elements in a repeating pattern. All previously reported membrane antennas were passive antennas; this is the first active membrane antenna that includes transmitting/receiving (T/R) electronic circuits as integral parts. Other integral parts of the antenna include a network of radio-frequency (RF) feed lines (more specifically, a corporate feed network) and of bias and control lines, all in the form of flexible copper strip conductors on flexible polymeric membranes. Each unit cell of a prototype antenna (see Figure 1) contains a patch antenna element and a compact T/R module that is compatible with flexible membrane circuitry. There are two membrane layers separated by a 12.7-mm air gap. Each membrane layer is made from a commercially available flexible circuit material that, as supplied, comprises a 127-micron-thick polyimide dielectric layer clad on both sides with 17.5-micron-thick copper layers. The copper layers are patterned into RF, bias, and control conductors. The T/R module is located on the back side of the ground plane and is RF-coupled to the patch element via a slot. The T/R module is a hybrid multilayer module assembled and packaged independently and attached to the membrane array. At the time of reporting the information for

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

    NASA Technical Reports Server (NTRS)

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

    2005-01-01

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

  3. Active polishing technology for large aperture aspherical mirror and ultra thin mirror

    NASA Astrophysics Data System (ADS)

    Cui, Xaingqun; Gao, Bilie; Li, Xinnan

    2006-02-01

    Some results on active polishing technology for large aperture aspherical mirrors and ultra thin mirrors, which have been developed in recent years in Nanjing Institute of Astronomical Optics and Technology, CAS, are presented in this paper. There are two polishing methods developed for the large aperture ultra thin mirrors with two different trial mirrors respectively. One is a hexagonal mirror with diagonal size of 1100mm, and thickness of 25mm by no-separate support method specially for polish the sub-mirror of Schmidt corrector of LAMOST, which is a national large scientific project of China. Another is a circular mirror with 1035mm in diameter and 26mm in thickness by active support method. The active stressed polishing technology developed for large aperture aspherical mirror with fast f ratio, and a paraboloidal mirror with a diameter of 910mm and an f ratio 2 as was successfully polished. The computer controlled polishing is also different from the normal way in the system. Some complicated aspects were added. The results showed the final surface accuracy of all these trial mirrors is better than expected requirements for normal application in astronomical telescopes.

  4. Horizon: A Proposal for Large Aperture, Active Optics in Geosynchronous Orbit

    NASA Technical Reports Server (NTRS)

    Chesters, Dennis; Jenstrom, Del

    2000-01-01

    In 1999, NASA's New Millennium Program called for proposals to validate new technology in high-earth orbit for the Earth Observing-3 (NMP EO3) mission to fly in 2003. In response, we proposed to test a large aperture, active optics telescope in geosynchronous orbit. This would flight-qualify new technologies for both Earth and Space science: 1) a future instrument with LANDSAT image resolution and radiometric quality watching continuously from geosynchronous station, and 2) the Next Generation Space Telescope (NGST) for deep space imaging. Six enabling technologies were to be flight-qualified: 1) a 3-meter, lightweight segmented primary mirror, 2) mirror actuators and mechanisms, 3) a deformable mirror, 4) coarse phasing techniques, 5) phase retrieval for wavefront control during stellar viewing, and 6) phase diversity for wavefront control during Earth viewing. Three enhancing technologies were to be flight- validated: 1) mirror deployment and latching mechanisms, 2) an advanced microcontroller, and 3) GPS at GEO. In particular, two wavefront sensing algorithms, phase retrieval by JPL and phase diversity by ERIM International, were to sense optical system alignment and focus errors, and to correct them using high-precision mirror mechanisms. Active corrections based on Earth scenes are challenging because phase diversity images must be collected from extended, dynamically changing scenes. In addition, an Earth-facing telescope in GEO orbit is subject to a powerful diurnal thermal and radiometric cycle not experienced by deep-space astronomy. The Horizon proposal was a bare-bones design for a lightweight large-aperture, active optical system that is a practical blend of science requirements, emerging technologies, budget constraints, launch vehicle considerations, orbital mechanics, optical hardware, phase-determination algorithms, communication strategy, computational burdens, and first-rate cooperation among earth and space scientists, engineers and managers

  5. Research of active panel technology for large aperture millimeter-wave/sub-millimeter-wave telescope

    NASA Astrophysics Data System (ADS)

    Wu, Xuhao; Cui, Xiangqun

    2010-05-01

    As Large Sky Area Multi-Object Fiber Spectroscopic Telescope (LAMOST) project was completed successfully, indicating the key technology of active optics has been mastered by the Chinese astronomical community, experts of Nanjing Institute of Astronomical Optics and Technology (NIAOT), builders of this project, started to consider how to use the technology developed in large optical telescope such as LAMOST to improve the performance of millimeterwave / sub-millimeter-wave telescope. In order to do more research work about active optics of millimeter submillimeter band and improve the performance of Delingha 13.7m millimeter-wave telescope, researchers of NIAOT intend to upgrade the reflect panel accuracy of this telescope. This paper will introduce the preliminary work of the accuracy-upgrading task, numerical simulation of the 13.7m telescope. In this presentation, the primary reflector finite element model (FEM) construction, gravity and thermal deformation, and modal analyze are described. The result shows that the gravity and thermal distortion of the reflector are contributed mostly by the back-structure and the active support for the panels is very necessary to restrain this kind of distortion.

  6. Large aperture diffractive space telescope

    DOEpatents

    Hyde, Roderick A.

    2001-01-01

    A large (10's of meters) aperture space telescope including two separate spacecraft--an optical primary objective lens functioning as a magnifying glass and an optical secondary functioning as an eyepiece. The spacecraft are spaced up to several kilometers apart with the eyepiece directly behind the magnifying glass "aiming" at an intended target with their relative orientation determining the optical axis of the telescope and hence the targets being observed. The objective lens includes a very large-aperture, very-thin-membrane, diffractive lens, e.g., a Fresnel lens, which intercepts incoming light over its full aperture and focuses it towards the eyepiece. The eyepiece has a much smaller, meter-scale aperture and is designed to move along the focal surface of the objective lens, gathering up the incoming light and converting it to high quality images. The positions of the two space craft are controlled both to maintain a good optical focus and to point at desired targets which may be either earth bound or celestial.

  7. Large aperture scanning airborne lidar

    NASA Technical Reports Server (NTRS)

    Smith, J.; Bindschadler, R.; Boers, R.; Bufton, J. L.; Clem, D.; Garvin, J.; Melfi, S. H.

    1988-01-01

    A large aperture scanning airborne lidar facility is being developed to provide important new capabilities for airborne lidar sensor systems. The proposed scanning mechanism allows for a large aperture telescope (25 in. diameter) in front of an elliptical flat (25 x 36 in.) turning mirror positioned at a 45 degree angle with respect to the telescope optical axis. The lidar scanning capability will provide opportunities for acquiring new data sets for atmospheric, earth resources, and oceans communities. This completed facility will also make available the opportunity to acquire simulated EOS lidar data on a near global basis. The design and construction of this unique scanning mechanism presents exciting technological challenges of maintaining the turning mirror optical flatness during scanning while exposed to extreme temperatures, ambient pressures, aircraft vibrations, etc.

  8. the Large Aperture GRB Observatory

    SciTech Connect

    Bertou, Xavier

    2009-04-30

    The Large Aperture GRB Observatory (LAGO) aims at the detection of high energy photons from Gamma Ray Bursts (GRB) using the single particle technique (SPT) in ground based water Cherenkov detectors (WCD). To reach a reasonable sensitivity, high altitude mountain sites have been selected in Mexico (Sierra Negra, 4550 m a.s.l.), Bolivia (Chacaltaya, 5300 m a.s.l.) and Venezuela (Merida, 4765 m a.s.l.). We report on the project progresses and the first operation at high altitude, search for bursts in 6 months of preliminary data, as well as search for signal at ground level when satellites report a burst.

  9. Large aperture Fresnel telescopes/011

    SciTech Connect

    Hyde, R.A., LLNL

    1998-07-16

    At Livermore we`ve spent the last two years examining an alternative approach towards very large aperture (VLA) telescopes, one based upon transmissive Fresnel lenses rather than on mirrors. Fresnel lenses are attractive for VLA telescopes because they are launchable (lightweight, packagable, and deployable) and because they virtually eliminate the traditional, very tight, surface shape requirements faced by reflecting telescopes. Their (potentially severe) optical drawback, a very narrow spectral bandwidth, can be eliminated by use of a second (much smaller) chromatically-correcting Fresnel element. This enables Fresnel VLA telescopes to provide either single band ({Delta}{lambda}/{lambda} {approximately} 0.1), multiple band, or continuous spectral coverage. Building and fielding such large Fresnel lenses will present a significant challenge, but one which appears, with effort, to be solvable.

  10. Very Large Aperture Diffractive Space Telescope

    SciTech Connect

    Hyde, Roderick Allen

    1998-04-20

    A very large (10's of meters) aperture space telescope including two separate spacecraft--an optical primary functioning as a magnifying glass and an optical secondary functioning as an eyepiece. The spacecraft are spaced up to several kilometers apart with the eyepiece directly behind the magnifying glass ''aiming'' at an intended target with their relative orientation determining the optical axis of the telescope and hence the targets being observed. The magnifying glass includes a very large-aperture, very-thin-membrane, diffractive lens, e.g., a Fresnel lens, which intercepts incoming light over its full aperture and focuses it towards the eyepiece. The eyepiece has a much smaller, meter-scale aperture and is designed to move along the focal surface of the magnifying glass, gathering up the incoming light and converting it to high quality images. The positions of the two space craft are controlled both to maintain a good optical focus and to point at desired targets.

  11. A review of large aperture Schlieren photography technique

    NASA Astrophysics Data System (ADS)

    Xu, Song-bo; Xie, Yong-jun; Chen, Lei

    2016-01-01

    Schlieren photography is a visual process to display the flow of fluids of varying density. It is widely used in wind tunnel tests to photograph the flow of air around objects. To achieve schlieren images with high sensitivity and high resolution, and satisfy the requirements of the large-scale wind tunnel tests, it is urgent to develop schlieren photographers with large aperture primary mirrors. However, the application of large aperture primary mirrors may bring many challenges in the design of the schlieren system. First, the surface figure of large aperture primary mirrors is difficult to control so that the support structure may need more strategical design. Second, because the schlieren system works under some severe environments of the wind tunnel test including the air disturbance, wind-induced ground vibration and high ambient pressure, it has to withstand serious instability risks to ensure a good schlieren image quality. In this work, the current status of the development in the large aperture schlieren systems is reviewed. Several advanced methods, for example, active damping control technique, focal spot monitoring technique, 18-points whilffletree support technique, etc.., are introduced to deal with the challenges of the large aperture schlieren system. This work aims at improving the technical development of large aperture schlieren photographer, which may contribute to the acquisition of the high sensitive and high resolution schlieren images and the improvement of the testing capability in wind tunnel experiments.

  12. Eyeglass. 1. Very large aperture diffractive telescopes.

    PubMed

    Hyde, R A

    1999-07-01

    The Eyeglass is a very large aperture (25-100-m) space telescope consisting of two distinct spacecraft, separated in space by several kilometers. A diffractive lens provides the telescope s large aperture, and a separate, much smaller, space telescope serves as its mobile eyepiece. Use of a transmissive diffractive lens solves two basic problems associated with very large aperture space telescopes; it is inherently launchable (lightweight, packagable, and deployable) it and virtually eliminates the traditional, very tight surface shape tolerances faced by reflecting apertures. The potential drawback to use of a diffractive primary (very narrow spectral bandwidth) is eliminated by corrective optics in the telescope s eyepiece; the Eyeglass can provide diffraction-limited imaging with either single-band (Deltalambda/lambda approximately 0.1), multiband, or continuous spectral coverage. PMID:18323902

  13. Large aperture adaptive optics for intense lasers

    NASA Astrophysics Data System (ADS)

    Deneuville, François; Ropert, Laurent; Sauvageot, Paul; Theis, Sébastien

    2015-05-01

    ISP SYSTEM has developed a range of large aperture electro-mechanical deformable mirrors (DM) suitable for ultra short pulsed intense lasers. The design of the MD-AME deformable mirror is based on force application on numerous locations thanks to electromechanical actuators driven by stepper motors. DM design and assembly method have been adapted to large aperture beams and the performances were evaluated on a first application for a beam with a diameter of 250mm at 45° angle of incidence. A Strehl ratio above 0.9 was reached for this application. Simulations were correlated with measurements on optical bench and the design has been validated by calculation for very large aperture (up to Ø550mm). Optical aberrations up to Zernike order 5 can be corrected with a very low residual error as for actual MD-AME mirror. Amplitude can reach up to several hundreds of μm for low order corrections. Hysteresis is lower than 0.1% and linearity better than 99%. Contrary to piezo-electric actuators, the μ-AME actuators avoid print-through effects and they permit to keep the mirror shape stable even unpowered, providing a high resistance to electro-magnetic pulses. The MD-AME mirrors can be adapted to circular, square or elliptical beams and they are compatible with all dielectric or metallic coatings.

  14. Study on test metrology of large aperture optical system wavefront

    NASA Astrophysics Data System (ADS)

    Liu, Zhiying; Fu, Yuegang; Gao, Tianyuan; Wang, Zhijian

    2009-05-01

    Large aperture optical system test has been a key problem for a long time. It could be solved by sub-aperture stitching method after the sub-apertures are tested. Sub-aperture stitching technology is a feasible method for testing large diameter optical system with small diameter interferometer sub-aperture stitching. Auto-collimating component will be needed with interferometer stitching method. Auto-collimating component is defined that the image could be kept stable when the optical component rotates about any axis in space. And the beam could be back along original optical path. By this means, auto collimation could be realized. The auto-collimating component is smaller than the test system. The whole wavefront of large aperture system could be tested through the method that the auto-collimating component moves along the guide rail and rotates about optical axis. A right angle roof prism is chosen as the auto-collimating component due to its character of easier manufacture. The active matrix, characteristic orientation and extreme axial is deduced with dynamic optics. The sub-aperture stitching testing process is simulated by ZEMAX in detail. The test result by stitching method is compared with that by directive test method for large aperture optical system. It is shown that the relative test error is less than 4.3λ 0/00. The sub -aperture stitching test method is verified.

  15. Large Aperture, Scanning, L-Band SAR

    NASA Technical Reports Server (NTRS)

    Moussessian, Alina; Del Castillo, Linda; Bach, Vinh; Grando, Maurio; Quijano, Ubaldo; Smith, Phil; Zawadzki, Mark

    2011-01-01

    We have developed the first L-band membrane-based active phased array. The antenna is a 16x16 element patch array with dimensions of 2.3mx2.6m. The array uses membrane-compatible Transmit/Receive (T/R) modules for electronic beam steering. We will discuss the antenna design, the fabrication of this large array, the T/R module development, the signal distribution approach and the measured results of the array.

  16. A large aperture electro-optic deflector

    NASA Astrophysics Data System (ADS)

    Bosco, A.; Boogert, S. T.; Boorman, G. E.; Blair, G. A.

    2009-05-01

    An electro-optic laser beam deflector with a clear optical aperture of 8.6 mm has been designed, realized, and tested. The electro-optic material used to implement the device was a MgO:LiNbO3 crystal. The exceptionally large aperture makes the device suitable for applications where fast scanning of high power laser beams is needed. The measured deflection angle was 120 μrad/kV for a total length of electro-optic material of 90 mm. A mode quality analysis of the laser beam revealed that the M2 of the laser is affected by less than 4% during scan operation when maximum driving voltage is applied.

  17. Design of large aperture focal plane shutter

    NASA Astrophysics Data System (ADS)

    Hu, Jia-wen; Ma, Wen-li; Huang, Jin-long

    2012-09-01

    To satisfy the requirement of large telescope, a large aperture focal plane shutter with aperture size of φ200mm was researched and designed to realize, which could be started and stopped in a relative short time with precise position, and also the blades could open and close at the same time at any orientation. Timing-belts and stepper motors were adopted as the drive mechanism. Velocity and position of the stepper motors were controlled by the PWM pulse generated by DSP. Exponential curve is applied to control the velocity of the stepper motors to make the shutter start and stop in a short time. The closing/open time of shutter is 0.2s, which meets the performance requirements of large telescope properly.

  18. The COronal Solar Magnetism Observatory (COSMO) Large Aperture Coronagraph

    NASA Astrophysics Data System (ADS)

    Tomczyk, Steve; Gallagher, Dennis; Wu, Zhen; Zhang, Haiying; Nelson, Pete; Burkepile, Joan; Kolinksi, Don; Sutherland, Lee

    2013-04-01

    The COSMO is a facility dedicated to observing coronal and chromospheric magnetic fields. It will be located on a mountaintop in the Hawaiian Islands and will replace the current Mauna Loa Solar Observatory (MLSO). COSMO will provide unique observations of the global coronal magnetic fields and its environment to enhance the value of data collected by other observatories on the ground (e.g. SOLIS, BBO NST, Gregor, ATST, EST, Chinese Giant Solar Telescope, NLST, FASR) and in space (e.g. SDO, Hinode, SOHO, GOES, STEREO, Solar-C, Solar Probe+, Solar Orbiter). COSMO will employ a fleet of instruments to cover many aspects of measuring magnetic fields in the solar atmosphere. The dynamics and energy flow in the corona are dominated by magnetic fields. To understand the formation of CMEs, their relation to other forms of solar activity, and their progression out into the solar wind requires measurements of coronal magnetic fields. The large aperture coronagraph, the Chromospheric and Prominence Magnetometer and the K-Coronagraph form the COSMO instrument suite to measure magnetic fields and the polarization brightness of the low corona used to infer electron density. The large aperture coronagraph will employ a 1.5 meter fuse silica singlet lens, birefringent filters, and a spectropolarimeter to cover fields of view of up to 1 degree. It will observe the corona over a wide range of emission lines from 530.3 nm through 1083.0 nm allowing for magnetic field measurements over a wide range of coronal temperatures (e.g. FeXIV at 530.3 nm, Fe X at 637.4 nm, Fe XIII at 1074.7 and 1079.8 nm. These lines are faint and require the very large aperture. NCAR and NSF have provided funding to bring the large aperture coronagraph to a preliminary design review state by the end of 2013. As with all data from Mauna Loa, the data products from COSMO will be available to the community via the Mauna Loa website: http://mlso.hao.ucar.edu

  19. Design of large aperture, low mass vacuum windows

    SciTech Connect

    Leonhardt, W.J.; Mapes, M.

    1993-01-01

    Large vacuum vessels are employed downstream of fixed targets in High Energy Physics experiments to provide a long path for particles to traverse without interacting with air molecules. These vessels generally have a large aperture opening known as a vacuum window which employs a thin membrane to preserve the vacuum environment yet allows the particles to pass through with a minimal effect on them. Several large windows have been built using a composite of Kevlar/Mylar including circular windows to a diameter of 96.5 cm and rectangular windows up to 193 cm x 86 cm. This paper describes the design, fabrication, testing and operating experience with these windows and relates the actual performance to theoretical predictions.

  20. Design of large aperture, low mass vacuum windows

    SciTech Connect

    Leonhardt, W.J.; Mapes, M.

    1993-07-01

    Large vacuum vessels are employed downstream of fixed targets in High Energy Physics experiments to provide a long path for particles to traverse without interacting with air molecules. These vessels generally have a large aperture opening known as a vacuum window which employs a thin membrane to preserve the vacuum environment yet allows the particles to pass through with a minimal effect on them. Several large windows have been built using a composite of Kevlar/Mylar including circular windows to a diameter of 96.5 cm and rectangular windows up to 193 cm x 86 cm. This paper describes the design, fabrication, testing and operating experience with these windows and relates the actual performance to theoretical predictions.

  1. Development of large aperture composite adaptive optics

    NASA Astrophysics Data System (ADS)

    Kmetik, Viliam; Vitovec, Bohumil; Jiran, Lukas; Nemcova, Sarka; Zicha, Josef; Inneman, Adolf; Mikulickova, Lenka; Pavlica, Richard

    2015-01-01

    Large aperture composite adaptive optics for laser applications is investigated in cooperation of Institute of Plasma Physic, Department of Instrumentation and Control Engineering FME CTU and 5M Ltd. We are exploring opportunity of a large-size high-power-laser deformable-mirror production using a lightweight bimorph actuated structure with a composite core. In order to produce a sufficiently large operational free aperture we are developing new technologies for production of flexible core, bimorph actuator and deformable mirror reflector. Full simulation of a deformable-mirrors structure was prepared and validated by complex testing. A deformable mirror actuation and a response of a complicated structure are investigated for an accurate control of the adaptive optics. An original adaptive optics control system and a bimorph deformable mirror driver were developed. Tests of material samples, components and sub-assemblies were completed. A subscale 120 mm bimorph deformable mirror prototype was designed, fabricated and thoroughly tested. A large-size 300 mm composite-core bimorph deformable mirror was simulated and optimized, fabrication of a prototype is carried on. A measurement and testing facility is modified to accommodate large sizes optics.

  2. Low-Cost Large Aperture Telescopes for Optical Communications

    NASA Technical Reports Server (NTRS)

    Hemmati, Hamid

    2006-01-01

    Low-cost, 0.5-1 meter ground apertures are required for near-Earth laser communications. Low-cost ground apertures with equivalent diameters greater than 10 meters are desired for deep-space communications. This presentation focuses on identifying schemes to lower the cost of constructing networks of large apertures while continuing to meet the requirements for laser communications. The primary emphasis here is on the primary mirror. A slumped glass spherical mirror, along with passive secondary mirror corrector and active adaptive optic corrector show promise as a low-cost alternative to large diameter monolithic apertures. To verify the technical performance and cost estimate, development of a 1.5-meter telescope equipped with gimbal and dome is underway.

  3. A Future Large-Aperture UVOIR Space Observatory: Study Overview

    NASA Astrophysics Data System (ADS)

    Postman, Marc; Thronson, Harley A.; Feinberg, Lee; Redding, David; Stahl, H. Philip

    2015-01-01

    The scientific drivers for very high angular resolution coupled with very high sensitivity and wavefront stability in the UV and optical wavelength regime have been well established. These include characterization of exoplanets in the habitable zones of solar type stars, probing the physical properties of the circumgalactic medium around z < 2 galaxies, and resolving stellar populations across a broad range of galactic environments. The 2010 NRC Decadal Survey and the 2013 NASA Science Mission Directorate 30-Year Roadmap identified a large-aperture UVOIR observatory as a priority future space mission. Our joint NASA GSFC/JPL/MSFC/STScI team has extended several earlier studies of the technology and engineering requirements needed to design and build a single filled aperture 10-meter class space-based telescope that can enable these ambitious scientific observations. We present here an overview of our new technical work including a brief summary of the reference science drivers as well as in-depth investigations of the viable telescope architectures, the requirements on thermal control and active wavefront control systems, and the range of possible launch configurations.

  4. Eyeglass: A Very Large Aperture Diffractive Space Telescope

    SciTech Connect

    Hyde, R; Dixit, S; Weisberg, A; Rushford, M

    2002-07-29

    Eyeglass is a very large aperture (25-100 meter) space telescope consisting of two distinct spacecraft, separated in space by several kilometers. A diffractive lens provides the telescope's large aperture, and a separate, much smaller, space telescope serves as its mobile eyepiece. Use of a transmissive diffractive lens solves two basic problems associated with very large aperture space telescopes; it is inherently fieldable (lightweight and flat, hence packagable and deployable) and virtually eliminates the traditional, very tight, surface shape tolerances faced by reflecting apertures. The potential drawback to use of a diffractive primary (very narrow spectral bandwidth) is eliminated by corrective optics in the telescope's eyepiece. The Eyeglass can provide diffraction-limited imaging with either single-band, multiband, or continuous spectral coverage. Broadband diffractive telescopes have been built at LLNL and have demonstrated diffraction-limited performance over a 40% spectral bandwidth (0.48-0.72 {micro}m). As one approach to package a large aperture for launch, a foldable lens has been built and demonstrated. A 75 cm aperture diffractive lens was constructed from 6 panels of 1 m thick silica; it achieved diffraction-limited performance both before and after folding. This multiple panel, folding lens, approach is currently being scaled-up at LLNL. We are building a 5 meter aperture foldable lens, involving 72 panels of 700 {micro}m thick glass sheets, diffractively patterned to operate as coherent f/50 lens.

  5. Fabrication and applications of large aperture diffractive optics

    SciTech Connect

    Dixit, S; Britten, J B; Hyde, R; Rushford, M; Summers, L; Toeppen, J

    2002-02-19

    Large aperture diffractive optics are needed in high power laser applications to protect against laser damage during operation and in space applications to increase the light gathering power and consequently the signal to noise. We describe the facilities we have built for fabricating meter scale diffractive optics and discuss several examples of these.

  6. Temperature characteristic of 808nm VCSELs with large aperture

    NASA Astrophysics Data System (ADS)

    Feng, Yuan; Feng, Dawei; Hao, Yongqin; Wang, Yong; Yan, Changling; Lu, Peng; Li, Yang

    2015-03-01

    In order to study the output characteristics of 808nm vertical cavity surface emitting laser(VCSEL) with large aperture at different temperature, 808nm VCSEL with 500μm emitting diameter are fabricated with Reticular Electrode Structure(RES). Lasing wavelength, optical power and the threshold current are measured by changing the temperature of heat sink. And an output power of 0.42W is achieved at 1.3A at room temperature under continuous wave operation. The central wavelength is 803.32nm, and the full width at half maximum is 0.16nm, the temperature shift is 0.06nm/°, the thermal resistance is 0.098°/mW. The testing results show that 808nm VCSEL with large aperture is good temperature characteristic.

  7. Mission definition for a large-aperture microwave radiometer spacecraft

    NASA Technical Reports Server (NTRS)

    Keafer, L. S., Jr.

    1981-01-01

    An Earth-observation measurements mission is defined for a large-aperture microwave radiometer spacecraft. This mission is defined without regard to any particular spacecraft design concept. Space data application needs, the measurement selection rationale, and broad spacecraft design requirements and constraints are described. The effects of orbital parameters and image quality requirements on the spacecraft and mission performance are discussed. Over the land the primary measurand is soil moisture; over the coastal zones and the oceans important measurands are salinity, surface temperature, surface winds, oil spill dimensions and ice boundaries; and specific measurement requirements have been selected for each. Near-all-weather operation and good spatial resolution are assured by operating at low microwave frequencies using an extremely large aperture antenna in a low-Earth-orbit contiguous mapping mode.

  8. Zinc selenide-based large aperture photo-controlled deformable mirror.

    PubMed

    Quintavalla, Martino; Bonora, Stefano; Natali, Dario; Bianco, Andrea

    2016-06-01

    Realization of large aperture deformable mirrors with a large density of actuators is important in many applications, and photo-controlled deformable mirrors (PCDMs) represent an innovation. Herein we show that PCDMs are scalable realizing a 2-inch aperture device based on a polycrystalline zinc selenide (ZnSe) as the photoconductive substrate and a thin polymeric reflective membrane. ZnSe is electrically characterized and analyzed through a model that we previously introduced. The PCDM is then optically tested, demonstrating its capabilities in adaptive optics. PMID:27244417

  9. Extracting spatial information from large aperture exposures of diffuse sources

    NASA Technical Reports Server (NTRS)

    Clarke, J. T.; Moos, H. W.

    1981-01-01

    The spatial properties of large aperture exposures of diffuse emission can be used both to investigate spatial variations in the emission and to filter out camera noise in exposures of weak emission sources. Spatial imaging can be accomplished both parallel and perpendicular to dispersion with a resolution of 5-6 arc sec, and a narrow median filter running perpendicular to dispersion across a diffuse image selectively filters out point source features, such as reseaux marks and fast particle hits. Spatial information derived from observations of solar system objects is presented.

  10. Large aperture spatial heterodyne imaging spectrometer: Principle and experimental results

    NASA Astrophysics Data System (ADS)

    Xiangli, Bin; Cai, Qisheng; Du, Shusong

    2015-12-01

    A large aperture spatial heterodyne imaging spectrometer (LASHIS) is proposed. It is a kind of pushbroom Fourier transform ultraspectral imager with no moving parts. This imaging spectrometer, based on a Sagnac lateral shearing interferometer combined with a pair of gratings, has the advantages of high spectral resolution, high throughput and robustness. The principle of LASHIS and its spectral retrieval method are introduced. The processing chain to convert raw images to ultraspectral datacube is also described. Experimental results demonstrate the high resolving power of LASHIS with the emission spectrum of a low pressure sodium lamp.

  11. Operational aspects of the Main Injector large aperture quadrupole (WQB)

    SciTech Connect

    Chou, W.; Bartelson, L.; Brown, B.; Capista, D.; Crisp, J.; DiMarco, J.; Fitzgerald, J.; Glass, H.; Harding, D.; Johnson, D.; Kashikhin, V.; /Fermilab

    2007-06-01

    A two-year Large Aperture Quadrupole (WQB) Project was completed in the summer of 2006 at Fermilab. [1] Nine WQBs were designed, fabricated and bench-tested by the Technical Division. Seven of them were installed in the Main Injector and the other two for spares. They perform well. The aperture increase meets the design goal and the perturbation to the lattice is minimal. The machine acceptance in the injection and extraction regions is increased from 40{pi} to 60{pi} mm-mrad. This paper gives a brief report of the operation and performance of these magnets. Details can be found in Ref [2].

  12. Large-aperture interferometer using local reference beam

    NASA Technical Reports Server (NTRS)

    Howes, W. L.

    1982-01-01

    A large-aperture interferometer was devised by adding a local-reference-beam-generating optical system to a schlieren system. Two versions of the interferometer are demonstrated, one employing 12.7 cm (5 in.) diameter schlieren optics, the other employing 30.48 cm (12 in.) diameter parabolic mirrors in an off-axis system. In the latter configuration a cylindrical lens is introduced near the light source to correct for astigmatism. A zone plate is a satisfactory decollimating element in the reference-beam arm of the interferometer. Attempts to increase the flux and uniformity of irradiance in the reference beam by using a diffuser are discussed.

  13. Silicon Powder Filters for Large-Aperture Cryogenic Receivers

    NASA Astrophysics Data System (ADS)

    Boone, Fletcher; Essinger-Hileman, T.; Bennett, C. L.; Marriage, T.; Xu, Z.

    2014-01-01

    Upcoming experiments probing for the existence of B-mode polarization in the cosmic microwave background (CMB) will require large arrays of background-limited detectors. This will necessitate the use of cryogenic receivers with large-aperture vacuum windows and correspondingly large low-pass infrared-blocking filters to minimize thermal load. Large-diameter filters composed of absorptive dielectrics are difficult to conductively cool adequately, and thus tend to heat up and re-radiate towards the focal plane. Reflective metal-mesh filters are challenging to manufacture at such large apertures and with feature sizes small enough to effectively block 300K thermal radiation. In order to overcome these difficulties, we have developed a novel type of thermal filter that scatters, rather than reflects or absorbs, unwanted infrared radiation. Comprised of ultra-pure silicon powder distributed within a polymethylpentene (PMP) substrate, these filters are not absorptive in the infrared while being transparent to microwaves, and are comparatively straightforward to produce. By adjusting the size of the silicon particles, the frequency cut-off of these low-pass filters is fully tunable. Small scale (70mm diameter, 3mm thickness) prototypes have exhibited <10% transmission throughout the infrared spectrum and <1% transmission at the peak of the 300K blackbody spectrum, while maintaining an estimated 97% transmission in the microwave regime.

  14. Optimal strategy for fabrication of large aperture aspheric surfaces.

    PubMed

    Feng, Yunpeng; Cheng, Haobo; Wang, Tan; Dong, Zhichao; Tam, Hon-Yuen

    2014-01-01

    Aspheric surfaces are widely used because of their desirable characteristics. Such a surface can obtain nearly perfect imaging quality with fewer optical elements and reduce the size and mass of optical systems. Various machine systems have been developed based on modern deterministic polishing technologies for large aperture aspheric surfaces. Several factors affect the final precision of large aperture aspheric surfaces, such as the velocity limit of the machine and the path design. Excess velocity, which will be truncated automatically by the computer numerical control system, may cause the dwell time to deviate from the desired time. When a path designed on a two-dimensional surface map with equidistant pitch is projected onto an aspheric surface, the pitch changes as a result of the varied curvature of the aspheric surface. This may affect the removal map and cause some ripple errors. A multiregion distribution strategy, which includes velocity checking, is proposed in this study to avoid exceeding the velocity limits. The strategy can be used to modify local errors and edge effects. A three-dimensional spiral path generation method is also presented using an iterative method to ensure uniformity in the space length of the adjacent circle of the spiral path. This process can reduce the ripple error caused by the overlapping of tool paths. A polishing experiment was conducted, and the results proved the validity of the proposed strategies. PMID:24514001

  15. Self-Referencing Hartmann Test for Large-Aperture Telescopes

    NASA Technical Reports Server (NTRS)

    Korechoff, Robert P.; Oseas, Jeffrey M.

    2010-01-01

    A method is proposed for end-to-end, full aperture testing of large-aperture telescopes using an innovative variation of a Hartmann mask. This technique is practical for telescopes with primary mirrors tens of meters in diameter and of any design. Furthermore, it is applicable to the entire optical band (near IR, visible, ultraviolet), relatively insensitive to environmental perturbations, and is suitable for ambient laboratory as well as thermal-vacuum environments. The only restriction is that the telescope optical axis must be parallel to the local gravity vector during testing. The standard Hartmann test utilizes an array of pencil beams that are cut out of a well-corrected wavefront using a mask. The pencil beam array is expanded to fill the full aperture of the telescope. The detector plane of the telescope is translated back and forth along the optical axis in the vicinity of the nominal focal plane, and the centroid of each pencil beam image is recorded. Standard analytical techniques are then used to reconstruct the telescope wavefront from the centroid data. The expansion of the array of pencil beams is usually accomplished by double passing the beams through the telescope under test. However, this requires a well-corrected, autocollimation flat, the diameter or which is approximately equal to that of the telescope aperture. Thus, the standard Hartmann method does not scale well because of the difficulty and expense of building and mounting a well-corrected, large aperture flat. The innovation in the testing method proposed here is to replace the large aperture, well-corrected, monolithic autocollimation flat with an array of small-aperture mirrors. In addition to eliminating the need for a large optic, the surface figure requirement for the small mirrors is relaxed compared to that required of the large autocollimation flat. The key point that allows this method to work is that the small mirrors need to operate as a monolithic flat only with regard to

  16. Large-aperture, high-damage-threshold optics for beamlet

    SciTech Connect

    Campbell, J.H.; Atherton, L.J.; DeYoreo, J.J.

    1996-06-01

    Beamlet serves as a test bed for the proposed National Ignition Facility (NIF) laser design and components. Therefore, its optics are similar in size and quality to those proposed for the NIF. In general, the optics in the main laser cavity and transport section of Beamlet are larger and have higher damage thresholds than the optics manufactured for any of the previous laser systems. In addition, the quality of the Beamlet optical materials is higher, leading to better wavefront quality, higher optical transmission, and lower-intensity modulation of the output laser beam than, for example, that typically achieved on Nova. In this article, the authors discuss the properties and characteristics of the large-aperture optics used on Beamlet.

  17. Optical simulation of large aperture spatial heterodyne imaging spectrometer

    NASA Astrophysics Data System (ADS)

    Cai, Qisheng; Xiangli, Bin; Fang, Yu

    2016-05-01

    Large aperture spatial heterodyne imaging spectrometer (LASHIS) is a new pushbroom Fourier transform ultraspectral imager with no moving parts. It is based on a Sagnac interferometer combined with a pair of gratings. In this paper, the basic principle of LASHIS is reviewed and an optical LASHIS model is set up in ZEMAX. Three interference images are presented, one is calculated according to the basic theory, one is simulated using the optical model in ZEMAX, and the other is generated by the experimental device set up in our laboratory. These three interference images show a good agreement with each other that demonstrate the correctness of the optical model. Using this model, we can simulate the interference image quickly. This image gives a visualized evaluation of the system performance, and it will be more convenient for system design or tolerance analysis of LASHIS.

  18. Advances in optical materials for large aperture lasers

    SciTech Connect

    Stokowski, S.E.; Lowdermilk, W.H.; Marchi, F.T.; Swain, J.E.; Wallerstein, E.P.; Wirtenson, G.R.

    1981-12-15

    Lawrence Livermore National Laboratory (LLNL) is using large aperture Nd: glass lasers to investigate the feasibility of inertial confinement fusion. In our experiments high power laser light is focussed onto a small (100 to 500 micron) target containing a deuterium-tritium fuel mixture. During the short (1 to 5 ns) laser pulse the fuel is compressed and heated, resulting in fusion reactions. The generation and control of the powerful laser pulses for these experiments is a challenging scientific and engineering task, which requires the development of new optical materials, fabrication techniques, and coatings. LLNL with the considerable cooperation and support from the optical industry, where most of the research and development and almost all the manufacturing is done, has successfully applied several new developments in these areas.

  19. Large-aperture interferometer with local reference beam

    NASA Technical Reports Server (NTRS)

    Howes, W. L.

    1984-01-01

    A large-aperture interferometer was devised by adding a local-reference-beam-generating optical system to a schlieren system. Two versions of the interferometer are demonstrated, one employing 12.7 cm (5 in.) diameter schlieren optics, the other employing 30.48 cm (12 in.) diameter parabolic mirrors in an off-axis system. In the latter configuration a cylindrical lens is introduced near the light source to correct for astigmatism. A zone plate is a satisfactory decollimating element in the reference-beam arm of the interferometer. Attempts to increase the flux and uniformity of irradiance in the reference beam by using a diffuser are discussed. Previously announced in STAR as N83-13979

  20. A Future Large-Aperture UVOIR Space Observatory: Reference Designs

    NASA Technical Reports Server (NTRS)

    Thronson, Harley; Rioux, Norman; Feinberg, Lee; Stahl, H. Philip; Redding, Dave; Jones, Andrew; Sturm, James; Collins, Christine; Liu, Alice

    2015-01-01

    Our joint NASA GSFC/JPL/MSFC/STScI study team has used community-provided science goals to derive mission needs, requirements, and candidate mission architectures for a future large-aperture, non-cryogenic UVOIR space observatory. We describe the feasibility assessment of system thermal and dynamic stability for supporting coronagraphy. The observatory is in a Sun-Earth L2 orbit providing a stable thermal environment and excellent field of regard. Reference designs include a 36-segment 9.2 m aperture telescope that stows within a five meter diameter launch vehicle fairing. Performance needs developed under the study are traceable to a variety of reference designs including options for a monolithic primary mirror.

  1. A future large-aperture UVOIR space observatory: reference designs

    NASA Astrophysics Data System (ADS)

    Rioux, Norman; Thronson, Harley; Feinberg, Lee; Stahl, H. Philip; Redding, Dave; Jones, Andrew; Sturm, James; Collins, Christine; Liu, Alice

    2015-09-01

    Our joint NASA GSFC/JPL/MSFC/STScI study team has used community-provided science goals to derive mission needs, requirements, and candidate mission architectures for a future large-aperture, non-cryogenic UVOIR space observatory. We describe the feasibility assessment of system thermal and dynamic stability for supporting coronagraphy. The observatory is in a Sun-Earth L2 orbit providing a stable thermal environment and excellent field of regard. Reference designs include a 36-segment 9.2 m aperture telescope that stows within a five meter diameter launch vehicle fairing. Performance needs developed under the study are traceable to a variety of reference designs including options for a monolithic primary mirror.

  2. Factors affecting the performance of large-aperture microphone arrays.

    PubMed

    Silverman, Harvey F; Patterson, William R; Sachar, Joshua

    2002-05-01

    Large arrays of microphones have been proposed and studied as a possible means of acquiring data in offices, conference rooms, and auditoria without requiring close-talking microphones. When such an array essentially surrounds all possible sources, it is said to have a large aperture. Large-aperture arrays have attractive properties of spatial resolution and signal-to-noise enhancement. This paper presents a careful comparison of theoretical and measured performance for an array of 256 microphones using simple delay-and-sum beamforming. This is the largest currently functional, all digital-signal-processing array that we know of. The array is wall-mounted in the moderately adverse environment of a general-purpose laboratory (8 m x 8 m x 3 m). The room has a T60 reverberation time of 550 ms. Reverberation effects in this room severely impact the array's performance. However, the width of the main lobe remains comparable to that of a simplified prediction. Broadband spatial resolution shows a single central peak with 10 dB gain about 0.4 m in diameter at the -3 dB level. Away from that peak, the response is approximately flat over most of the room. Optimal weighting for signal-to-noise enhancement degrades the spatial resolution minimally. Experimentally, we verify that signal-to-noise gain is less than proportional to the square root of the number of microphones probably due to the partial correlation of the noise between channels, to variation of signal intensity with polar angle about the source, and to imperfect correlation of the signal over the array caused by reverberations. We show measurements of the relative importance of each effect in our environment. PMID:12051434

  3. Factors affecting the performance of large-aperture microphone arrays

    NASA Astrophysics Data System (ADS)

    Silverman, Harvey F.; Patterson, William R.; Sachar, Joshua

    2002-05-01

    Large arrays of microphones have been proposed and studied as a possible means of acquiring data in offices, conference rooms, and auditoria without requiring close-talking microphones. When such an array essentially surrounds all possible sources, it is said to have a large aperture. Large-aperture arrays have attractive properties of spatial resolution and signal-to-noise enhancement. This paper presents a careful comparison of theoretical and measured performance for an array of 256 microphones using simple delay-and-sum beamforming. This is the largest currently functional, all digital-signal-processing array that we know of. The array is wall-mounted in the moderately adverse environment of a general-purpose laboratory (8 m×8 m×3 m). The room has a T60 reverberation time of 550 ms. Reverberation effects in this room severely impact the array's performance. However, the width of the main lobe remains comparable to that of a simplified prediction. Broadband spatial resolution shows a single central peak with 10 dB gain about 0.4 m in diameter at the -3 dB level. Away from that peak, the response is approximately flat over most of the room. Optimal weighting for signal-to-noise enhancement degrades the spatial resolution minimally. Experimentally, we verify that signal-to-noise gain is less than proportional to the square root of the number of microphones probably due to the partial correlation of the noise between channels, to variation of signal intensity with polar angle about the source, and to imperfect correlation of the signal over the array caused by reverberations. We show measurements of the relative importance of each effect in our environment.

  4. A feasibility study into the screening and imaging of hand luggage for threat items at 35 GHz using an active large aperture (1.6 m) security screening imager

    NASA Astrophysics Data System (ADS)

    Bowring, Nicholas J.; O'Reilly, Dean; Salmon, Neil A.; Andrews, David A.; Rezgui, Nacer-Ddine; Harmer, Stuart W.

    2013-10-01

    The feasibility of screening hand luggage for concealed threat items such as Person-Borne Improvised Explosive Devices (PBIED's) both metallic and non-metallic, together with handguns and at millimetre wavelengths is investigated. Previous studies by the authors and others indicate that hand baggage material and fabric is much more transmissive and has less scattering at lower millimetre wave frequencies and the ability to use K-band active imaging with high spatial resolution presents an opportunity to image and hence recognise concealed threats. For this feasibility study, a 1.6 m aperture, 35 GHz security screening imaging system with a spatial resolution of 2.5 cm and a depth of field of around 5 cm is employed, using spatially incoherent illuminating panels to enhance image contrast. In this study, realistic scenarios using backpacks containing a realistic range of threat and non-threat items are scanned, both carried and standalone. This range of items contains large vessels suitable for containing simulated home-made PBIED's and handguns. The comprehensive list of non-threat items includes laptops, bottles, clothing and power supplies. For this study, the range at which imaging data at standoff distances can be acquired is confined to that of the particular system in use, although parameters such as illumination and integration time are optimised. However, techniques for extrapolating towards effective standoff distances using aperture synthesis imagers are discussed. The transmission loss through fabrics and clothing that may form, or be contained in baggage, are reported over range of frequencies ranging from 26 to 110 GHz.

  5. Error analysis of large aperture static interference imaging spectrometer

    NASA Astrophysics Data System (ADS)

    Li, Fan; Zhang, Guo

    2015-12-01

    Large Aperture Static Interference Imaging Spectrometer is a new type of spectrometer with light structure, high spectral linearity, high luminous flux and wide spectral range, etc ,which overcomes the contradiction between high flux and high stability so that enables important values in science studies and applications. However, there're different error laws in imaging process of LASIS due to its different imaging style from traditional imaging spectrometers, correspondingly, its data processing is complicated. In order to improve accuracy of spectrum detection and serve for quantitative analysis and monitoring of topographical surface feature, the error law of LASIS imaging is supposed to be learned. In this paper, the LASIS errors are classified as interferogram error, radiometric correction error and spectral inversion error, and each type of error is analyzed and studied. Finally, a case study of Yaogan-14 is proposed, in which the interferogram error of LASIS by time and space combined modulation is mainly experimented and analyzed, as well as the errors from process of radiometric correction and spectral inversion.

  6. BLAST: The Balloon-Borne Large Aperture Submillimeter Telescope

    NASA Technical Reports Server (NTRS)

    Devlin, Mark; Ade, Peter; Bock, Jamie; Dicker, Simon; Griffin, Matt; Gunderson, Josh; Halpern, Mark; Hargrave, Peter; Hughes, David; Klein, Jeff

    2004-01-01

    BLAST is the Balloon-borne Large-Aperture Sub-millimeter Telescope. It will fly from a Long Duration Balloon (LDB) platform from Antarctica. The telescope design incorporates a 2 m primary mirror with large-format bolometer arrays operating at 250, 350 and 500 microns. By providing the first sensitive large-area (10 sq. deg.) sub-mm surveys at these wavelengths, BLAST will address some of the most important galactic and cosmological questions regarding the formation and evolution of stars, galaxies and clusters. Galactic and extragalactic BLAST surveys will: (1) identify large numbers of high-redshift galaxies; (2) measure photometric redshifts, rest-frame FIR luminosities and star formation rates thereby constraining the evolutionary history of the galaxies that produce the FIR and sub-mm background; (3) measure cold pre-stellar sources associated with the earliest stages of star and planet formation; (4) make high-resolution maps of diffuse galactic emission over a wide range of galactic latitudes. In addition to achieving the above scientific goals, the exciting legacy of the BLAST LDB experiment will be a catalogue of 3000-5000 extragalactic sub-mm sources and a 100 sq. deg. sub-mm galactic plane survey. Multi-frequency follow-up observations from SIRTF, ASTRO-F, and Herschel, together with spectroscopic observations and sub-arcsecond imaging from ALMA are essential to understand the physical nature of the BLAST sources.

  7. The Balloon-borne Large Aperture Submillimeter Telescope: BLAST

    NASA Astrophysics Data System (ADS)

    Pascale, E.; Ade, P. A. R.; Bock, J. J.; Chapin, E. L.; Chung, J.; Devlin, M. J.; Dicker, S.; Griffin, M.; Gundersen, J. O.; Halpern, M.; Hargrave, P. C.; Hughes, D. H.; Klein, J.; MacTavish, C. J.; Marsden, G.; Martin, P. G.; Martin, T. G.; Mauskopf, P.; Netterfield, C. B.; Olmi, L.; Patanchon, G.; Rex, M.; Scott, D.; Semisch, C.; Thomas, N.; Truch, M. D. P.; Tucker, C.; Tucker, G. S.; Viero, M. P.; Wiebe, D. V.

    2008-07-01

    The Balloon-borne Large Aperture Submillimeter Telescope (BLAST) is a suborbital surveying experiment designed to study the evolutionary history and processes of star formation in local galaxies (including the Milky Way) and galaxies at cosmological distances. The BLAST continuum camera, which consists of 270 detectors distributed between three arrays, observes simultaneously in broadband (30%) spectral windows at 250, 350, and 500 μm. The optical design is based on a 2 m diameter telescope, providing a diffraction-limited resolution of 30'' at 250 μm. The gondola pointing system enables raster mapping of arbitrary geometry, with a repeatable positional accuracy of ~30''; postflight pointing reconstruction to lesssim5'' rms is achieved. The onboard telescope control software permits autonomous execution of a preselected set of maps, with the option of manual override. In this paper we describe the primary characteristics and measured in-flight performance of BLAST. BLAST performed a test flight in 2003 and has since made two scientifically productive long-duration balloon flights: a 100 hr flight from ESRANGE (Kiruna), Sweden to Victoria Island, northern Canada in 2005 June; and a 250 hr, circumpolar flight from McMurdo Station, Antarctica, in 2006 December.

  8. The Balloon-borne Large Aperture Submillimeter Telescope: BLAST

    NASA Astrophysics Data System (ADS)

    Semisch, Christopher; BLAST Collaboration

    2007-12-01

    The Balloon-borne Large Aperture Submillimeter Telescope (BLAST) is a sub-orbital survey-experiment designed to study the evolutionary history and processes of star formation in local galaxies (including the Milky Way) and galaxies at cosmological distances. The BLAST continuum camera, which consists of 270 detectors distributed between 3 arrays, observes simultaneously in broad-band (30%) spectral-windows at 250µm, 350µm, and 500µm. The optical design is based on a 2m diameter Cassegrain telescope, providing a diffraction-limited resolution of 30" at 250µm. The gondola pointing system enables raster-like maps of arbitrary geometry, with a repeatable positional accuracy of 30"; post-flight pointing reconstruction to < 5" rms is also achieved. The on-board telescope control software permits autonomous execution of a pre-selected set of maps, with the option of manual intervention. Since a test-flight in 2003, BLAST has made two scientifically productive long-duration balloon flights: a 100-hour flight from ESRANGE (Kiruna), Sweden to Victoria Island, northern Canada in June 2005, and a 250-hour, circumpolar-flight from McMurdo Station, Antarctica, in December 2006.

  9. Design considerations for a large aperture high field superconducting dipole

    SciTech Connect

    Harfoush, F.; Ankenbrandt, C.; Harrison, M.; Kerby, J.; Koepke, K.; Mantsch, P.; Nicol, T.; Riddiford, A.; Theilacker, J.

    1989-03-01

    The final phase of the Fermilab upgrade proposal calls for a new ring of superconducting magnets to be placed in the existing Main Accelerator tunnel. The goal of this design study is to specify a high field dipole (HFD) that is capable of supporting fixed target operation (ramping, resonant extraction) at a field of 6.6T (1.5 Tev) and colliding beam physics at 8.0T (1.8 Tev). The magnetic field quality at high field is set by the large amplitude orbits associated with resonant extraction. The field quality must therefore be at least as good as the existing Tevatron magnets which fulfill these criteria. The high fields and large aperture of this magnet result in large forces on the coil and collar assemblies. Therefore, the cold mass design must be able to sustain these forces while providing sufficient cooling to the coils during 4.2 K fixed target operation, and a minimum heat load during 1.8 K collider operation. The design work is still in progress but a cosine-theta, cold-iron dipole with a 70mm inner diameter coil has been tentatively adopted. This report presents details on the conductor and cable parameters, coil cross-section, projected manufacturing tolerances, iron yoke design, and cold mass assembly. 4 refs., 5 figs., 1 tab.

  10. Phase retrieval in situ measurement for large aperture parabolic mirror

    NASA Astrophysics Data System (ADS)

    Ding, Lingyan; Wu, Yulie; Li, Shengyi; Liao, Yang; Shu, Yong

    2010-10-01

    Phase retrieval is a promising method for in-situ metrology and has been applied to spherical mirror surface metrology successfully. To meet the requirement of in-situ measurement in manufacturing large aperture parabolic mirror, a new method using phase retrieval technology is developed. In this method, an approximately parallel beam is used to illuminate the large parabolic mirror. The beam is produced by a point light source far away from the tested mirror. Then, intensity of diffraction patterns near the focus is measured by CCD. The experiment of testing a parabolic mirror with aperture 400mm and radius of curvature at vertex 2789.7mm is described. And some advices of improving the setup are presented. Errors brought by the approximately parallel beam are compensated by an algorithm derived from GS iterative algorithm. Phase retrieval result is consistent with that measured by interferometer sub-aperture stitching in error distribution, PV value and RMS value. The experiment shows that this method features simple optical path, good anti-vibration ability and acceptable accuracy.

  11. Hybrid Electrostatic/Flextensional Mirror for Lightweight, Large-Aperture, and Cryogenic Space Telescopes

    NASA Technical Reports Server (NTRS)

    Patrick, Brian; Moore, James; Hackenberger, Wesley; Jiang, Xiaoning

    2013-01-01

    A lightweight, cryogenically capable, scalable, deformable mirror has been developed for space telescopes. This innovation makes use of polymer-based membrane mirror technology to enable large-aperture mirrors that can be easily launched and deployed. The key component of this innovation is a lightweight, large-stroke, cryogenic actuator array that combines the high degree of mirror figure control needed with a large actuator influence function. The latter aspect of the innovation allows membrane mirror figure correction with a relatively low actuator density, preserving the lightweight attributes of the system. The principal components of this technology are lightweight, low-profile, high-stroke, cryogenic-capable piezoelectric actuators based on PMN-PT (piezoelectric lead magnesium niobate-lead titanate) single-crystal configured in a flextensional actuator format; high-quality, low-thermal-expansion polymer membrane mirror materials developed by NeXolve; and electrostatic coupling between the membrane mirror and the piezoelectric actuator assembly to minimize problems such as actuator print-through.

  12. The Balloon-borne Large Aperture Submillimeter Telescope: BLAST

    NASA Astrophysics Data System (ADS)

    Truch, Matthew D. P.; Ade, P. A. R.; Bock, J. J.; Chapin, E. L.; Chung, J.; Devlin, M. J.; Dicker, S.; Griffin, M.; Gundersen, J. O.; Halpern, M.; Hargrave, P. C.; Hughes, D. H.; Klein, J.; MacTavish, C. J.; Marsden, G.; Martin, P. G.; Martin, T. G.; Mauskopf, P.; Netterfield, C. B.; Olmi, L.; Pascale, E.; Patanchon, G.; Rex, M.; Scott, D.; Semisch, C.; Thomas, N. E.; Tucker, C.; Tucker, G. S.; Viero, M. P.; Wiebe, D. V.

    2009-01-01

    The Balloon-borne Large Aperture Submillimeter Telescope (BLAST) is a suborbital surveying experiment designed to study the evolutionary history and processes of star formation in local galaxies (including the Milky Way) and galaxies at cosmological distances. The BLAST continuum camera, which consists of 270 detectors distributed between three arrays, observes simultaneously in broadband (30%) spectral windows at 250, 350, and 500 microns. The optical design is based on a 2 m diameter telescope, providing a diffraction-limited resolution of 30" at 250 microns. The gondola pointing system enables raster mapping of arbitrary geometry, with a repeatable positional accuracy of 30"; postflight pointing reconstruction to <5" rms is achieved. The onboard telescope control software permits autonomous execution of a preselected set of maps, with the option of manual override. On this poster, we describe the primary characteristics and measured in-flight performance of BLAST. BLAST performed a test flight in 2003 and has since made two scientifically productive long-duration balloon flights: a 100 hour flight from ESRANGE (Kiruna), Sweden to Victoria Island, northern Canada in 2005 June; and a 250 hour, circumpolar flight from McMurdo Station, Antarctica in 2006 December. The BLAST collaboration acknowledges the support of NASA through grants NAG5-12785, NAG5-13301, and NNGO-6GI11G, the Canadian Space Agency (CSA), the Science and Technology Facilities Council (STFC), Canada's Natural Sciences and Engineering Research Council (NSERC), the Canada Foundation for Innovation, the Ontario Innovation Trust, the Puerto Rico Space Grant Consortium, the Fondo Institucional para la Investigacion of the University of Puerto Rico, and the National Science Foundation Office of Polar Programs.

  13. NST: Thermal Modeling for a Large Aperture Solar Telescope

    NASA Astrophysics Data System (ADS)

    Coulter, Roy

    2011-05-01

    Late in the 1990s the Dutch Open Telescope demonstrated that internal seeing in open, large aperture solar telescopes can be controlled by flushing air across the primary mirror and other telescope structures exposed to sunlight. In that system natural wind provides a uniform air temperature throughout the imaging volume, while efficiently sweeping heated air away from the optics and mechanical structure. Big Bear Solar Observatory's New Solar Telescope (NST) was designed to realize that same performance in an enclosed system by using both natural wind through the dome and forced air circulation around the primary mirror to provide the uniform air temperatures required within the telescope volume. The NST is housed in a conventional, ventilated dome with a circular opening, in place of the standard dome slit, that allows sunlight to fall only on an aperture stop and the primary mirror. The primary mirror is housed deep inside a cylindrical cell with only minimal openings in the side at the level of the mirror. To date, the forced air and cooling systems designed for the NST primary mirror have not been implemented, yet the telescope regularly produces solar images indicative of the absence of mirror seeing. Computational Fluid Dynamics (CFD) analysis of the NST primary mirror system along with measurements of air flows within the dome, around the telescope structure, and internal to the mirror cell are used to explain the origin of this seemingly incongruent result. The CFD analysis is also extended to hypothetical systems of various scales. We will discuss the results of these investigations.

  14. Experimental instrumentation system for the Phased Array Mirror Extendible Large Aperture (PAMELA) test program

    NASA Technical Reports Server (NTRS)

    Boykin, William H., Jr.

    1993-01-01

    Adaptive optics are used in telescopes for both viewing objects with minimum distortion and for transmitting laser beams with minimum beam divergence and dance. In order to test concepts on a smaller scale, NASA MSFC is in the process of setting up an adaptive optics test facility with precision (fraction of wavelengths) measurement equipment. The initial system under test is the adaptive optical telescope called PAMELA (Phased Array Mirror Extendible Large Aperture). Goals of this test are: assessment of test hardware specifications for PAMELA application and the determination of the sensitivities of instruments for measuring PAMELA (and other adaptive optical telescopes) imperfections; evaluation of the PAMELA system integration effort and test progress and recommended actions to enhance these activities; and development of concepts and prototypes of experimental apparatuses for PAMELA.

  15. Large aperture tunable-focus liquid lens using shape memory alloy spring.

    PubMed

    Hasan, Nazmul; Kim, Hanseup; Mastrangelo, Carlos H

    2016-06-13

    A tunable-focus large aperture liquid lens is constructed using shape memory alloy (SMA) springs as actuators. The lens mainly consists of a shallow liquid-filled cylindrical cavity bound by a thin compressible annular rim and encapsulated by a flexible circular membrane on the top of the rim and a rigid circular plate at the rim bottom. The lens optical power is adjusted by a controlled compression of the annular rim via actuation of the three shape-memory alloy (SMA) springs. Since the volume of the cavity liquid is constant, the rim compression bulges the flexible membrane outward thus reducing its radius of curvature and the lens focal length. The fabricated tunable lens demonstrated an optical power range of 0-4 diopters utilizing a driving voltage less than 3V. Lens optical wavefront profiling was done using a Shack-Hartmann sensor displaying a RMS wave front error of 0.77 µm and 1.68 µm at 0 D and + 4 D. The aperture diameter and thickness of the fabricated lens are 34 mm and 9 mm, respectively, while weighing 16.7 g. PMID:27410350

  16. MOIRE: ground demonstration of a large aperture diffractive transmissive telescope

    NASA Astrophysics Data System (ADS)

    Atcheson, Paul; Domber, Jeanette; Whiteaker, Kevin; Britten, Jerald A.; Dixit, Shamasundar N.; Farmer, Brandon

    2014-08-01

    The desire to field space-based telescopes with apertures in excess of 10 meter diameter is forcing the development of extreme lightweighted large optomechanical structures. Sparse apertures, shell optics, and membrane optics are a few of the approaches that have been investigated and demonstrated. Membrane optics in particular have been investigated for many years. The MOIRE approach in which the membrane is used as a transmissive diffractive optical element (DOE) offers a significant relaxation in the control requirements on the membrane surface figure, supports extreme lightweighting of the primary collecting optic, and provides a path for rapid low cost production of the primary optical elements. Successful development of a powered meter-scale transmissive membrane DOE was reported in 2012. This paper presents initial imaging results from integrating meter-scale transmissive DOEs into the primary element of a 5- meter diameter telescope architecture. The brassboard telescope successfully demonstrates the ability to collect polychromatic high resolution imagery over a representative object using the transmissive DOE technology. The telescope includes multiple segments of a 5-meter diameter telescope primary with an overall length of 27 meters. The object scene used for the demonstration represents a 1.5 km square complex ground scene. Imaging is accomplished in a standard laboratory environment using a 40 nm spectral bandwidth centered on 650 nm. Theoretical imaging quality for the tested configuration is NIIRS 2.8, with the demonstration achieving NIIRS 2.3 under laboratory seeing conditions. Design characteristics, hardware implementation, laboratory environmental impacts on imagery, image quality metrics, and ongoing developments will be presented.

  17. Intracavity Herriott-cell testbed for large-aperture femtosecond optics

    NASA Astrophysics Data System (ADS)

    Rácz, Péter; Nagy, Benedek J.; Ferencz, Kárpát; Dombi, Péter

    2014-12-01

    We demonstrate a versatile test method of large-aperture femtosecond mirrors inside a long-cavity Ti:sapphire laser oscillator. The Herriott-cell inside the cavity is utilized for the purpose of carrying out full-aperture testing. The method is highly sensitive to the homogeneity of mirror reflectivity and group delay dispersion over the whole mirror surface and it is suitable for testing both high reflector and chirped mirrors starting from 2” size up to arbitrarily large apertures.

  18. Active membrane phased array radar

    NASA Technical Reports Server (NTRS)

    Moussessian, Alina; Del Castillo, Linda; Huang, John; Sadowy, Greg; Hoffman, James; Smith, Phil; Hatake, Toshiro; Derksen, Chuck; Lopez, Bernardo; Caro, Ed

    2005-01-01

    We have developed the first membrane-based active phased array in L-band (1.26GHz). The array uses membrane compatible Transmit/Receive (T/R) modules (membrane T/R) for each antenna element. We use phase shifters within each T/R module for electronic beam steering. We will discuss the T/R module design and integration with the membrane, We will also present transmit and receive beam-steering results for the array.

  19. APPLICATION OF LARGE APERTURE EMATS TO WELD INSPECTION

    SciTech Connect

    Maclauchlan, D. T.; Clark, S. P.; Hancock, J. W.

    2008-02-28

    One of the most significant developments in EMAT operation is the incorporation of phased array techniques. Phased array EMATs enable electronic beam steering and focusing while operating with temporally short pulses for good range resolution. Using phased array EMAT operation, multiple high powered pulsers are combined in the generation of the ultrasonic wave and multiple elements are combined in the reception of the ultrasonic wave, for improved sensitivity. EMATs make it practical to operate with shear horizontal (SH) waves and scan over a metal part's surface. An EMAT generated line force at the surface launches shear horizontal waves with uniform amplitude for beam angles from -90 deg. to 90 deg. Shear horizontal waves also reflect without mode conversion from surfaces that are parallel to the polarization of the shear wave displacements. The combination of these advantages makes phased array EMATs well suited for weld inspection. Recently, BWXT Services has developed a 32 active channel EMAT phased array system for operation up to 5 MHz. In addition, each element can be constructed with several sub-elements, alternating in polarity, to effectively multiply the number of active elements for a restricted range of beam angles. For example by using elements comprised of 4 sub elements, a 128 active element aperture designed for operation with a nominal 60 deg. beam angle provides good beam steering and focusing performance for 45 deg. to 70 deg. beam angles. The large active apertures allow the use of highly focused beams for good defect detection and high resolution imaging of weld defects. Application of this system to weld inspections has verified that good defect detection and imaging is possible. In addition, operation with SH waves has proven to provide improved detection of lack of fusion at the cap and root of the weld for certain weld geometries. The system has also been used to demonstrate the inspection of submerged metal arc welds while welding.

  20. Structural Feasibility Analysis of a Robotically Assembled Very Large Aperture Optical Space Telescope

    NASA Technical Reports Server (NTRS)

    Wilkie, William Keats; Williams, R. Brett; Agnes, Gregory S.; Wilcox, Brian H.

    2007-01-01

    This paper presents a feasibility study of robotically constructing a very large aperture optical space telescope on-orbit. Since the largest engineering challenges are likely to reside in the design and assembly of the 150-m diameter primary reflector, this preliminary study focuses on this component. The same technology developed for construction of the primary would then be readily used for the smaller optical structures (secondary, tertiary, etc.). A reasonable set of ground and on-orbit loading scenarios are compiled from the literature and used to define the structural performance requirements and size the primary reflector. A surface precision analysis shows that active adjustment of the primary structure is required in order to meet stringent optical surface requirements. Two potential actuation strategies are discussed along with potential actuation devices at the current state of the art. The finding of this research effort indicate that successful technology development combined with further analysis will likely enable such a telescope to be built in the future.

  1. Performance of large-aperture optical switches for high-energy inertial-confinement fusion lasers

    SciTech Connect

    Rhodes, M.A.; Woods, B.; DeYoreo, J.J.; Roberts, D.; Atherton, L.J.

    1995-08-20

    We describe the design and performance of large-aperture ({lt}30 cm {times} 30 cm) optical switches that have demonstrated, for the first time to our knowledge, active switching of a high-energy ({lt}5 kJ) optical pulse in an inertial-confinement fusion laser. These optical switches, which consist of a plasma-electrode Pockels cell (PEPC) and a passive polarizer, permit the design of efficient, multipass laser amplifiers. In a PEPC, plasma discharges on the faces of a thin (1-cm) electro-optic crystal (KDP or KD{bold |}P) act as highly conductive and transparent electrodes. These plasma electrodes facilitate rapid ({lt}100 ns) and uniform charging of the crystal to the half-wave voltage and discharging back to 0 V. We discuss the operating principles, design, optical performance, and technical issues of a 32 cm {times} 32 cm prototype PEPC with both KDP and KD{bold |}P crystals, and a 37 cm {times} 37 cm PEPC with a KDP crystal for the Beamlet laser. This PEPC recently switched a 6-kJ, 3-ns pulse in a four-pass cavity.

  2. MICRONERVA: A Novel Approach to Large Aperture Astronomical Spectroscopy

    NASA Astrophysics Data System (ADS)

    Hall, Ryan; Plavchan, Peter; Geneser, Claire; Giddens, Frank; Spangler, Sophia

    2016-06-01

    MICRONERVA (MICRO Novel Exoplanet Radial Velocity Array) is a project to measure precise spectroscopic radial velocities. The cost of telescopes are a strong function of diameter, and light gathering power as opposed to angular resolution is the fundamental driver for telescope design for many spectroscopic science applications. By sacrificing angular resolution, many multiple smaller fiber-fed telescopes can be combined to synthesize the light gathering power of a larger diameter telescope at a lower effective cost. For our MICRONERVA prototype, based upon the larger MINERVA project, we will attempt to demonstrate that an array of four 8-inch CPC Celestron telescopes can be automated with sufficient active guiding precision for robust nightly robotic operations. The light from each telescope is coupled into single mode fibers, which are conveniently matched to the point spread function of 8-inch telescopes, which can be diffraction limited at red wavelengths in typical seeing at good observing sites. Additionally, the output from an array of single mode fibers provides stable output illumination of a spectrograph, which is a critical requirement of future precise radial velocity instrumentation. All of the hardware from the system is automated using Python programs and ASCOM and MaxIm DL software drivers. We will present an overview of the current status of the project and plans for future work. The detection of exoplanets using the techniques of MICRONERVA could potentially enable cost reductions for many types of spectroscopic research.

  3. Enzymatically active ultrathin pepsin membranes.

    PubMed

    Raaijmakers, Michiel J T; Schmidt, Thomas; Barth, Monika; Tutus, Murat; Benes, Nieck E; Wessling, Matthias

    2015-05-11

    Enzymatically active proteins enable efficient and specific cleavage reactions of peptide bonds. Covalent coupling of the enzymes permits immobilization, which in turn reduces autolysis-induced deactivation. Ultrathin pepsin membranes were prepared by facile interfacial polycondensation of pepsin and trimesoyl chloride. The pepsin membrane allows for simultaneous enzymatic conversion and selective removal of digestion products. The large water fluxes through the membrane expedite the transport of large molecules through the pepsin layers. The presented method enables the large-scale production of ultrathin, cross-linked, enzymatically active membranes. PMID:25779668

  4. Off-axis multipass amplifier as a large aperture driver stage for fusion lasers.

    PubMed

    Murray, J E; Downs, D C; Hunt, J T; Hermes, G L; Warren, W E

    1981-03-01

    A multipass amplifier configuration is described which has potential as a large aperture, high gain driver stage for fusion laser systems. We avoid the present limitations of large aperture switches by using an off-angle geometry that does not require an optical switch. The saturated gain characteristics of this multipass amplifier are optimized numerically. Three potential problems are investigated experimentally, self-lasing, output beam quality, and amplified spontaneous emission output. The results indicate comparable cost for comparable performance to a linear chain, with some operational advantage for the multipass driver stage. PMID:20309212

  5. π/K/p identification with a large-aperture ring-imaging cherenkov counter

    NASA Astrophysics Data System (ADS)

    Adams, M.; Bastin, A.; Coutrakon, G.; Glass, H.; Jaffe, D.; Kirz, J.; McCarthy, R.; Hubbard, J. R.; Mangeot, Ph.; Mullie, J.; Peisert, A.; Tichit, J.; Bouclier, R.; Charpak, G.; Santiard, J. C.; Sauli, F.; Crittenden, J.; Hsiung, Y.; Kaplan, D.; Brown, C.; Childress, S.; Finley, D.; Ito, A.; Jonckheere, A.; Jöstlein, H.; Lederman, L.; Orava, R.; Smith, S.; Sugano, K.; Ueno, K.; Maki, A.; Hemmi, Y.; Miyake, K.; Nakamura, T.; Sasao, N.; Sakai, Y.; Gray, R.; Plaag, R.; Rothberg, J.; Rutherfoord, J.; Young, K.

    1983-11-01

    The operating large aperture ring-imaging Cherenkov detector from the FNAL experiment E605 is described. Cherenkov ultraviolet photons are detected with a multi-step avalanche chamber using a He/TEA gas mixture and π/K/p separation is obtained from 50 to 200 GeV/ c.

  6. Imaging the midcontinent rift beneath Lake Superior using large aperture seismic data

    USGS Publications Warehouse

    Trehu, Anne M.; Morel-a-l'Huissier, Patrick; Meyer, R.; Hajnal, Z.; Karl, J.; Mereu, R. F.; Sexton, J.; Shay, J.; Chan, W. K.; Epili, D.; Jefferson, T.; Shih, X. R.; Wendling, S.; Milkereit, B.; Green, A.; Hutchinson, Deborah R.

    1991-01-01

    We present a detailed velocity model across the 1.1 billion year old Midcontinent Rift System (MRS) in central Lake Superior. The model was derived primarily from onshore-offshore large-aperture seismic and gravity data. High velocities obtained within a highly reflective half-graben that was imaged on coincident seismic reflection data demonstrate the dominantly mafic composition of the graben fill and constrain its total thickness to be at least 30km. Strong wide-angle reflections are observed from the lower crust and Moho, indicating that the crust is thickest (55–60km) beneath the axis of the graben. The total crustal thickness decreases rapidly to about 40 km beneath the south shore of the lake and decreases more gradually to the north. Above the Moho is a high-velocity lower crust interpreted to result from syn-rift basaltic intrusion into and/or underplating beneath the Archean lower crust. The lower crust is thickest beneath the axis of the main rift half-graben. A second region of thick lower crust is found approximately 100km north of the axis of the rift beneath a smaller half graben that is interpreted to reflect an earlier stage of rifting. The crustal model presented here resembles recent models of some passive continental margins and is in marked contrast to many models of both active and extinct Phanerozoic continental rift zones. It demonstrates that the Moho is a dynamic feature, since the pre-rift Moho is probably within or above the high-velocity lower crust, whereas the post-rift Moho is defined as the base of this layer. In the absence of major tectonic activity, however, the Moho is very stable, since the large, abrupt variations in crustal thickness beneath the MRS have been preserved for at least a billion years.

  7. Thermal Analysis of the Advanced Technology Large Aperture Space Telescope (ATLAST) 8 Meter Primary Mirror

    NASA Technical Reports Server (NTRS)

    Hornsby, Linda; Stahl, H. Philip; Hopkins, Randall C.

    2010-01-01

    The Advanced Technology Large Aperture Space Telescope (ATLAST) preliminary design concept consists of an 8 meter diameter monolithic primary mirror enclosed in an insulated, optical tube with stray light baffles and a sunshade. ATLAST will be placed in orbit about the Sun-Earth L2 and will experience constant exposure to the sun. The insulation on the optical tube and sunshade serve to cold bias the telescope which helps to minimize thermal gradients. The primary mirror will be maintained at 280K with an active thermal control system. The geometric model of the primary mirror, optical tube, sun baffles, and sunshade was developed using Thermal Desktop(R) SINDA/FLUINT(R) was used for the thermal analysis and the radiation environment was analyzed using RADCAD(R). A XX node model was executed in order to characterize the static performance and thermal stability of the mirror during maneuvers. This is important because long exposure observations, such as extra-solar terrestrial planet finding and characterization, require a very stable observatory wave front. Steady state thermal analyses served to predict mirror temperatures for several different sun angles. Transient analyses were performed in order to predict thermal time constant of the primary mirror for a 20 degree slew or 30 degree roll maneuver. This paper describes the thermal model and provides details of the geometry, thermo-optical properties, and the environment which influences the thermal performance. All assumptions that were used in the analysis are also documented. Parametric analyses are summarized for design parameters including primary mirror coatings and sunshade configuration. Estimates of mirror heater power requirements are reported. The thermal model demonstrates results for the primary mirror heated from the back side and edges using a heater system with multiple independently controlled zones.

  8. The development of large-aperture test system of infrared camera and visible CCD camera

    NASA Astrophysics Data System (ADS)

    Li, Yingwen; Geng, Anbing; Wang, Bo; Wang, Haitao; Wu, Yanying

    2015-10-01

    Infrared camera and CCD camera dual-band imaging system is used in many equipment and application widely. If it is tested using the traditional infrared camera test system and visible CCD test system, 2 times of installation and alignment are needed in the test procedure. The large-aperture test system of infrared camera and visible CCD camera uses the common large-aperture reflection collimator, target wheel, frame-grabber, computer which reduces the cost and the time of installation and alignment. Multiple-frame averaging algorithm is used to reduce the influence of random noise. Athermal optical design is adopted to reduce the change of focal length location change of collimator when the environmental temperature is changing, and the image quality of the collimator of large field of view and test accuracy are also improved. Its performance is the same as that of the exotic congener and is much cheaper. It will have a good market.

  9. Large aperture and polarizer-free liquid crystal lenses for ophthalmic applications

    NASA Astrophysics Data System (ADS)

    Lin, Yi-Hsin; Chen, Hung-Shan; Chang, Chia-Ming; Wang, Yu-Jen

    2014-10-01

    Large aperture and polarizer-free liquid crystal lenses (LC lenses) based on a double-layered structure for ophthalmic applications are demonstrated. The polarizer-free LC lens functions as both of a positive lens and a negative lens with large aperture size of 10mm. The lens power is electrically and continuously tunable ranging from -1.32 Diopter to 1.83 Diopter. To demonstrate the polarization independency, the wavefronts of the LC lenses under different polarized light were measured and discussed. The detail operations of the applied voltage and frequency are also discussed. The imaging performance of the LC lens is also evaluated. This study provide a detail understanding of the polarizer-free LC lenses based on a double-layered structure.

  10. Eyeglass Large Aperture, Lightweight Space Optics FY2000 - FY2002 LDRD Strategic Initiative

    SciTech Connect

    Hyde, R

    2003-02-10

    A series of studies by the Air Force, the National Reconnaissance Office and NASA have identified the critical role played by large optics in fulfilling many of the space related missions of these agencies. Whether it is the Next Generation Space Telescope for NASA, high resolution imaging systems for NRO, or beam weaponry for the Air Force, the diameter of the primary optic is central to achieving high resolution (imaging) or a small spot size on target (lethality). While the detailed requirements differ for each application (high resolution imaging over the visible and near-infrared for earth observation, high damage threshold but single-wavelength operation for directed energy), the challenges of a large, lightweight primary optic which is space compatible and operates with high efficiency are the same. The advantage of such large optics to national surveillance applications is that it permits these observations to be carried-out with much greater effectiveness than with smaller optics. For laser weapons, the advantage is that it permits more tightly focused beams which can be leveraged into either greater effective range, reduced laser power, and/or smaller on-target spot-sizes; weapon systems can be made either much more effective or much less expensive. This application requires only single-wavelength capability, but places an emphasis upon robust, rapidly targetable optics. The advantages of large aperture optics to astronomy are that it increases the sensitivity and resolution with which we can view the universe. This can be utilized either for general purpose astronomy, allowing us to examine greater numbers of objects in more detail and at greater range, or it can enable the direct detection and detailed examination of extra-solar planets. This application requires large apertures (for both light-gathering and resolution reasons), with broad-band spectral capability, but does not emphasize either large fields-of-view or pointing agility. Despite

  11. The Advanced Technology Large Aperture Space Telescope (ATLAST): Science Drivers and Technology Developments

    NASA Technical Reports Server (NTRS)

    Postman, Marc; Brown, Tom; Sembach, Kenneth; Giavalisco, Mauro; Traub, Wesley; Stapelfeldt, Karl; Calzetti, Daniela; Oegerle, William; Rich, R. Michael; Stahl, H. Phillip; Tumlinson, Jason; Mountain, Matt; Soummer, Remi; Hyde, Tupper

    2011-01-01

    The Advanced Technology Large-Aperture Space Telescope (ATLAST) is a concept for an 8-meter to 16-meter UVOIR space observatory for launch in the 2025-2030 era. ATLAST will allow astronomers to answer fundamental questions at the forefront of modern astrophysics, including "Is there life elsewhere in the Galaxy?" We present a range of science drivers and the resulting performance requirements for ATLAST (8 to 16 milliarcsecond angular resolution, diffraction limited imaging at 0.5 m wavelength, minimum collecting area of 45 square meters, high sensitivity to light wavelengths from 0.1 m to 2.4 m, high stability in wavefront sensing and control). We also discuss the priorities for technology development needed to enable the construction of ATLAST for a cost that is comparable to current generation observatory-class space missions. Keywords: Advanced Technology Large-Aperture Space Telescope (ATLAST); ultraviolet/optical space telescopes; astrophysics; astrobiology; technology development.

  12. Multidirectional curved integral imaging with large depth by additional use of a large-aperture lens.

    PubMed

    Shin, Dong-Hak; Lee, Byoungho; Kim, Eun-Soo

    2006-10-01

    We propose a curved integral imaging system with large depth achieved by the additional use of a large-aperture lens in a conventional large-depth integral imaging system. The additional large-aperture lens provides a multidirectional curvature effect and improves the viewing angle. The proposed system has a simple structure due to the use of well-fabricated, unmodified flat devices. To calculate the proper elemental images for the proposed system, we explain a modified computer-generated pickup technique based on an ABCD matrix and analyze an effective viewing zone in the proposed system. From experiments, we show that the proposed system has an improved viewing angle of more than 7 degrees compared with conventional integral imaging. PMID:16983427

  13. Testing the large aperture optical components by the sub-aperture stitching interferometer

    NASA Astrophysics Data System (ADS)

    He, Yong; Wang, Zhao-xuan; Wang, Qing; Ji, Bo

    2008-03-01

    Nowadays many large aperture optical components are widely used in the high-tech area, how to test them become more and more important. Here describes a new method to test the large aperture optical components using the small aperture interferometer, deduce how to get the aperture number and the concrete process of the stitching parameter in a systematic way, finally get the best plan to choose the sub-aperture of the square and circular optical plane. To specify the stability of the method we operate an experiment, the result shows that the stitching accuracy can reach λ/10, it meet the need of the inertia constraint fusion etc, that is good enough to be used in the high-tech area.

  14. Initial Technology Assessment for the Large-Aperture UV-Optical-Infrared (LUVOIR) Mission Concept Study

    NASA Technical Reports Server (NTRS)

    Bolcar, Matthew R.; Feinberg, Lee; France, Kevin; Rauscher, Bernard J.; Redding, David; Schiminovich, David

    2016-01-01

    The NASA Astrophysics Division's 30-Year Roadmap prioritized a future large-aperture space telescope operating in the ultra-violet/optical/infrared wavelength regime. The Association of Universities for Research in Astronomy envisioned a similar observatory, the High Definition Space Telescope. And a multi-institution group also studied the Advanced Technology Large Aperture Space Telescope. In all three cases, a broad science case is outlined, combining general astrophysics with the search for biosignatures via direct-imaging and spectroscopic characterization of habitable exoplanets. We present an initial technology assessment that enables such an observatory that is currently being studied for the 2020 Decadal Survey by the Large UV/Optical/Infrared (LUVOIR) surveyor Science and Technology Definition Team. We present here the technology prioritization for the 2016 technology cycle and define the required technology capabilities and current state-of-the-art performance. Current, planned, and recommended technology development efforts are also reported.

  15. High-power narrow-band terahertz generation using large-aperture photoconductors

    SciTech Connect

    Park, S.G.; Weiner, A.M.; Melloch, M.R. . School of Electrical and Computer Engineering); Siders, C.W.; Siders, J.L.W.; Taylor, A.J. )

    1999-08-01

    Large-aperture biased photoconductive emitters which can generate high-power narrow-band terahertz (THz) radiation are developed. These emitters avoid saturation at high fluence excitation and achieve enhanced peak power spectral density by employing a thick layer of short-lifetime low-temperature-grown GaAs (LT-GaAs) photoconductor and multiple-pulse excitation. THz waveforms are calculated from the saturation theory of large-aperture photoconductors, and a comparison is made between theory and measurement. A direct comparison of the multiple-pulse saturation properties of THz emission from semi-insulating GaAs and LT-GaAs emitters reveals a strong dependence on the carrier lifetime. In particular, the data demonstrate that saturation is avoided only when the interpulse spacing is longer than the carrier lifetime.

  16. Performance results for Beamlet: A large aperture multipass Nd glass laser

    SciTech Connect

    Campbell, J.H.; Barker, C.E.; VanWonterghem, B.M.; Speck, D.R.; Behrendt, W.C.; Murray, J.R.; Caird, J.A.; Decker, D.E.; Smith, I.C.

    1995-04-11

    The Beamlet laser is a large aperture, flashlamp pumped Nd: glass laser that is a scientific prototype of an advanced Inertial Fusion laser. Beamlet has achieved third harmonic, conversion efficiency of near 80% with its nominal 35cm {times} 35cm square beam at mean 3{omega} fluences in excess of 8 J/cm{sup 2}(3-ns). Beamlet uses an adaptive optics system to correct for aberrations and achieve less than 2 {times} diffraction limited far field spot size.

  17. Fabrication of large aperture kinoform phase plates in fused silica for smoothing focal plane intensity profiles

    SciTech Connect

    Rushford, M.; Dixit, S.; Thomas, I.; Perry, M.

    1996-04-26

    We have fabricated large aperture (40-cm) kinoform phase plates for producing super-Gaussian focal plane intensity profiles. The continuous phase screen, designed using a new iterative procedure, was fabricated in fused silica as a 16-level, one-wave deep rewrapped phase profile using a lithographic process and wet etching in buffered hydrofluoric acid. The observed far-field contains 94% of the incident energy inside the desired spot.

  18. Partial feedback unstable resonator on small scale supersonic large aperture chemical laser

    NASA Astrophysics Data System (ADS)

    Wang, Hongyan; Wang, Rui; Li, Lei

    2015-05-01

    There is always a challenge on large aperture medium power laser's resonator design, stable resonator would supports significant higher order transverse modes, folded and telescope stable resonator are too complex and not preferred by engineers, unstable resonator need rather large round trip gain to compensate its high geometric out-coupling, which is difficult for this kind of laser since its gain length is limited due to the power level and large aperture. Partial feedback unstable resonator had been proposed to tackle this difficulty since the early days of laser development, however, the debates of its effect never stopped even with those distinguished optical resonator scientists such as Siegman, Anan'ev, and Weber. Recently integrated partial feedback unstable resonator design had been successfully demonstrated on a medium size chemical oxygen iodine laser. In this paper, we carry this resonator configuration on a small scale discharge driven supersonic nozzle array Hydrogen Fluoride chemical laser, a typical large aperture short gain length device. With magnification equals 4/3, we successfully get ten Watts level ring beam output.

  19. Fast inspection of bulk and surface defects of large aperture optics in high power lasers

    NASA Astrophysics Data System (ADS)

    Zhao, Yuan'an; Hu, Guohang; Liu, Shijie; Yi, Kui; Shao, Jianda

    2015-05-01

    Laser induced damage for nanosecond pulse duration is attributed to the existence of defects. The growth and polishing, as well as coating deposition, may induce versatile kinds of defects, including dig, scratch and inclusion. It is special important to get the information of the defects, such as size and location, which is the basis to know the origin of the defects and figures out effective techniques to eliminate it. It is quite easy to get the information of the defects with micron-level resolution, but it is time-consuming and is not suitable for fast inspection of the large aperture (hundreds of millimeters). In this work, on-the-fly image capture technique was employed to realize fast inspection of large aperture optics. A continuous green laser was employed as illumination source to enhance and enlarge the image of bulk defects. So it could detect the submicron-scale defects. A transmission microscopy with white light illumination was employed to detect the surface defect. Its field of view was about 2.8mm×1.6mm. The sample was raster scanned driving by a stepper motor through the stationary illumination laser and digital camera, and the speed to scan the sample was about 10mm/s. The results of large aperture optics proved the functions of this fast inspection technique.

  20. Tracking marine mammals and ships with small and large-aperture hydrophone arrays

    NASA Astrophysics Data System (ADS)

    Gassmann, Martin

    Techniques for passive acoustic tracking in all three spatial dimensions of marine mammals and ships were developed for long-term acoustic datasets recorded continuously over months using custom-designed arrays of underwater microphones (hydrophones) with spacing ranging from meters to kilometers. From the three-dimensional tracks, the acoustical properties of toothed whales and ships, such as sound intensity and directionality, were estimated as they are needed for the passive acoustic abundance estimation of toothed whales and for a quantitative description of the contribution of ships to the underwater soundscape. In addition, the tracks of the toothed whales reveal their underwater movements and demonstrate the potential of the developed tracking techniques to investigate their natural behavior and responses to sound generated by human activity, such as from ships or military SONAR. To track the periodically emitted echolocation sounds of toothed whales in an acoustically refractive environment in the upper ocean, a propagation-model based technique was developed for a hydrophone array consisting of one vertical and two L-shaped subarrays deployed from the floating instrument platform R/P FLIP. The technique is illustrated by tracking a group of five shallow-diving killer whales showing coordinated behavior. The challenge of tracking the highly directional echolocation sounds of deep-diving (< 1 km) toothed whales, in particular Cuvier's beaked whales, was addressed by embedding volumetric small-aperture (≈ 1 m element spacing) arrays into a large-aperture (≈ 1 km element spacing) seafloor array to reduce the minimum number of required receivers from five to two. The capabilities of this technique are illustrated by tracking several groups of up to three individuals over time periods from 10 min to 33 min within an area of 20 km2 in the Southern California Bight. To track and measure the underwater radiated sound of ships, a frequency domain beamformer was

  1. X-ray refractive large aperture rolled prism lenses as condensers for x-ray tubes

    NASA Astrophysics Data System (ADS)

    Vogt, H.; Simon, M.; Last, A.; Marschall, F.; Mohr, J.; Nazmov, V.; Eisenhower, R.; Mettendorf, K. U.

    2011-10-01

    At the Institute of Microstructure Technology (IMT) at Karlsruhe Institute of Technology (KIT), refractive X-ray optics are developed. These optics are proposed to be used as condenser optics in X-Ray spectroscopy and microscopy applications with an X-ray tube as a source. To produce the lenses, a thin structured foil with equidistant fins in triangular form is casted from a structured silicon wafer. The foil is then wound around a glass fibre core. Due to this fabrication method, it is possible to produce large-aperture lenses with low absorption in comparison to other types of refractive X-Ray optics, like X-ray lenses with continuous parabolic shape or prism lenses. The first are limited due to their absorption while the latter are limited due to their mechanical stability of the prism columns. The optimisation of the so called X-Ray rolled prism lenses (RXPL) is underway at the institute and involves several parameters. One important property of the lenses is the correct form of the wound foil layers. This determines the number of necessary refractive elements at a given radius, which in turn determines the refracted slope and focal position of the transmitted beam. The spatial extent of the x-ray source is also being accounted for in the lens design. Another important point is the diameter of the winding core, which should be as small as possible due to the fact that the winding core reduces the active area of the lens. The rolling process itself is also revised to produce lenses with the above-mentioned small diameter winding cores and bend foil layers while sustaining a tight- fitting foil bundle. The lenses are studied at different energies and types of X-Ray tubes, as well as synchrotron sources, to gain additional information of the internal structure of the lens after the winding process. In this paper the current status of the lens development and results at X-Ray tube sources for use in diffractometers is presented.

  2. An Engineering Design Reference Mission for a Future Large-Aperture UVOIR Space Observatory

    NASA Astrophysics Data System (ADS)

    Thronson, Harley A.; Bolcar, Matthew R.; Clampin, Mark; Crooke, Julie A.; Redding, David; Rioux, Norman; Stahl, H. Philip

    2016-01-01

    From the 2010 NRC Decadal Survey and the NASA Thirty-Year Roadmap, Enduring Quests, Daring Visions, to the recent AURA report, From Cosmic Birth to Living Earths, multiple community assessments have recommended development of a large-aperture UVOIR space observatory capable of achieving a broad range of compelling scientific goals. Of these priority science goals, the most technically challenging is the search for spectroscopic biomarkers in the atmospheres of exoplanets in the solar neighborhood. Here we present an engineering design reference mission (EDRM) for the Advanced Technology Large-Aperture Space Telescope (ATLAST), which was conceived from the start as capable of breakthrough science paired with an emphasis on cost control and cost effectiveness. An EDRM allows the engineering design trade space to be explored in depth to determine what are the most demanding requirements and where there are opportunities for margin against requirements. Our joint NASA GSFC/JPL/MSFC/STScI study team has used community-provided science goals to derive mission needs, requirements, and candidate mission architectures for a future large-aperture, non-cryogenic UVOIR space observatory. The ATLAST observatory is designed to operate at a Sun-Earth L2 orbit, which provides a stable thermal environment and excellent field of regard. Our reference designs have emphasized a serviceable 36-segment 9.2 m aperture telescope that stows within a five-meter diameter launch vehicle fairing. As part of our cost-management effort, this particular reference mission builds upon the engineering design for JWST. Moreover, it is scalable to a variety of launch vehicle fairings. Performance needs developed under the study are traceable to a variety of additional reference designs, including options for a monolithic primary mirror.

  3. End-to-End Assessment of a Large Aperture Segmented Ultraviolet Optical Infrared (UVOIR) Telescope Architecture

    NASA Technical Reports Server (NTRS)

    Feinberg, Lee; Bolcar, Matt; Liu, Alice; Guyon, Olivier; Stark,Chris; Arenberg, Jon

    2016-01-01

    Key challenges of a future large aperture, segmented Ultraviolet Optical Infrared (UVOIR) Telescope capable of performing a spectroscopic survey of hundreds of Exoplanets will be sufficient stability to achieve 10-10 contrast measurements and sufficient throughput and sensitivity for high yield Exo-Earth spectroscopic detection. Our team has collectively assessed an optimized end to end architecture including a high throughput coronagraph capable of working with a segmented telescope, a cost-effective and heritage based stable segmented telescope, a control architecture that minimizes the amount of new technologies, and an Exo-Earth yield assessment to evaluate potential performance.

  4. Engineering Specification for Large-aperture UVO Space Telescopes Derived from Science Requirements

    NASA Technical Reports Server (NTRS)

    Stahl, H. Philip; Arnold, William; Bevan, Ryan M.; Smith, W. Scott.; Kirk, Charles S.; Postman, Mark

    2013-01-01

    An advanced large aperture UV/optical UVO space telescope is required for the next generation of astrophysics and exoplanet science. The science requirements of proposed exoplanet and astrophysics missions were used to determine the encircled energy, point spread function stability and thermal environment requirements. These requirements then determine the optical wavefront specification for potential telescope assemblies which can fit inside current and planned launch vehicles. The optical wavefront specification becomes the top level of the error budget that is split into various sources that control the structural, thermal and optical design.

  5. Development of an efficient large-aperture high damage-threshold sol-gel diffraction grating.

    SciTech Connect

    Ashley, Carol S.; Rambo, Patrick K.; Schwarz, Jens; Dunphy, Darren Robert; Branson, Eric D.; Smith, Ian Craig; Johnson, William Arthur; Reed, Scott T.; Cook, Adam W.

    2005-03-01

    In order to develop the next generation of high peak intensity lasers, new grating technology providing higher damage thresholds and large apertures is required. The current assumption is that this technical innovation will be multilayer dielectric gratings, wherein the uppermost layer of a thin film mirror is etched to create the desired binary phase grating. A variant of this is explored with the upper grating layer being a lower density gelatin-based volume phase grating in either sol-gel or dichromated gelatin. One key benefit is the elimination of the etching step.

  6. Spacecraft conceptual design for the 8-meter Advanced Technology Large Aperture Space Telescope (ATLAST)

    NASA Astrophysics Data System (ADS)

    Hopkins, Randall C.; Capizzo, Peter; Fincher, Sharon; Hornsby, Linda S.; Jones, David; Mosier, Gary; Stahl, H. Philip; Thomas, Dan; Thompson, Kevin S.

    2010-07-01

    The Advanced Concepts Office at Marshall Space Flight Center completed a brief spacecraft design study for the 8- meter monolithic Advanced Technology Large Aperture Space Telescope (ATLAST-8m). This spacecraft concept provides all power, communication, telemetry, avionics, guidance and control, and thermal control for the observatory, and inserts the observatory into a halo orbit about the second Sun-Earth Lagrange point. The multidisciplinary design team created a simple spacecraft design that enables component and science instrument servicing, employs articulating solar panels for help with momentum management, and provides precise pointing control while at the same time fast slewing for the observatory.

  7. Electro-optic harmonic conversion switch for large-aperture multipass laser systems

    SciTech Connect

    Henesian, M.A.; Goldhar, J.; Haas, R.A.

    1984-08-01

    The authors have demonstrated electro-optically tuned second-harmonic generation using Type I KDP inside a plasma-electrode discharge cell. An axial voltage of +/- 52 kV is required to switch a 1.064-..mu..m beam by conversion to 0.53 ..mu..m, in agreement with theory. Electro-optically tuned harmonic generation may be combined with a recently developed transparent plasma electrode to produce a large-aperture switch for multipass laser systems. 7 references, 4 figures, 1 table.

  8. Large aperture vibrating wire monitor with two mechanically coupled wires for beam halo measurements

    SciTech Connect

    Arutunian, S. G.; Avetisyan, A. E.; Davtyan, M. M.; Harutyunyan, G. S.; Vasiniuk, I. E.; Chung, M.; Scarpine, V.

    2014-03-01

    Development of a new type of Vibrating Wire Monitor (VWM), which has two mechanically coupled wires (vibrating and target), is presented. The new monitor has a much larger aperture size than the previous model of the VWM, and thus allows us to measure transverse beam halos more effectively. A prototype of such a large aperture VWM with a target wire length of 60 mm was designed, manufactured, and bench-tested. Initial beam measurements have been performed at the Fermilab High Intensity Neutrino Source (HINS) facility, and key results are presented.

  9. Spacecraft Conceptual Design for the 8-Meter Advanced Technology Large Aperture Space Telescope (ATLAST)

    NASA Technical Reports Server (NTRS)

    Hopkins, Randall C.; Capizzo, Peter; Fincher, Sharon; Hornsby, Linda S.; Jones, David

    2010-01-01

    The Advanced Concepts Office at Marshall Space Flight Center completed a brief spacecraft design study for the 8-meter monolithic Advanced Technology Large Aperture Space Telescope (ATLAST-8m). This spacecraft concept provides all power, communication, telemetry, avionics, guidance and control, and thermal control for the observatory, and inserts the observatory into a halo orbit about the second Sun-Earth Lagrange point. The multidisciplinary design team created a simple spacecraft design that enables component and science instrument servicing, employs articulating solar panels for help with momentum management, and provides precise pointing control while at the same time fast slewing for the observatory.

  10. Designs for a large-aperture telescope to map the CMB 10× faster.

    PubMed

    Niemack, Michael D

    2016-03-01

    Current large-aperture cosmic microwave background (CMB) telescopes have nearly maximized the number of detectors that can be illuminated while maintaining diffraction-limited image quality. The polarization-sensitive detector arrays being deployed in these telescopes in the next few years will have roughly 10⁴ detectors. Increasing the mapping speed of future instruments by at least an order of magnitude is important to enable precise probes of the inflationary paradigm in the first fraction of a second after the big bang and provide strong constraints on cosmological parameters. The CMB community has begun planning a next generation "Stage IV" CMB project that will be comprised of multiple telescopes with between 10⁵-10⁶ detectors to pursue these goals. This paper introduces the new crossed Dragone telescope and receiver optics designs that increase the usable diffraction-limited field-of-view, and therefore the mapping speed, by an order of magnitude compared to the upcoming generation of large-aperture instruments. Polarization systematics and engineering considerations are presented, including a preliminary receiver model to demonstrate that these designs will enable high efficiency illumination of >10⁵ detectors in a next generation CMB telescope. PMID:26974631

  11. Recent Enhancements of the Phased Array Mirror Extendible Large Aperture (PAMELA) Telescope Testbed at MSFC

    NASA Technical Reports Server (NTRS)

    Rakoczy, John; Montgomery, Edward E.; Lindner, Jeff

    2000-01-01

    Recent incremental upgrades to the Phased Array Mirror Extendible Large Aperture (PAMELA) telescope testbed have enabled the demonstration of phasing (with a monochromatic source) of clusters of primary mirror segments down to the diffraction limit. PAMELA upgrades include an improved Shack-Hartmann wavefront sensor, passive viscoelastic damping treatments for the voice-coil actuators, mechanical improvement of mirror surface figures, and optical bench baffling. This report summarizes the recent PAMELA upgrades, discusses the lessons learned, and presents a status of this unique testbed for wavefront sensing and control. The Marshall Space Flight Center acquired the Phased Array Mirror Extendible Large Aperture (PAMELA) telescope in 1993 after Kaman Aerospace was unable to complete integration and testing under the limited SDIO and DARPA funding. The PAMELA is a 36-segment, half-meter aperture, adaptive telescope which utilizes a Shack-Hartmann wavefront sensor, inductive coil edge sensors, voice coil actuators, imaging CCD cameras and interferometry for figure alignment, wavefront sensing and control. MSFC originally obtained the PAMELA to supplement its research in the interactions of control systems with flexible structures. In August 1994, complete tip, tilt and piston control was successfully demonstrated using the Shack-Hartmann wavefront sensor and the inductive edge sensors.

  12. Optimization analysis of primary mirror in large aperture telescope based on workbench

    NASA Astrophysics Data System (ADS)

    Feng, Zhengsen; Wang, Guomin

    2015-10-01

    With the diameter increasing for large aperture telescope primary mirror, the gravity caused by the increased of surface size will directly affect the quality of optical imaging, the adjustment of large aperture primary mirror will be frequent according to the requirement of observation. As the angle and the azimuth's transformation of primary mirror influences the surface shape accuracy immediately, the rational design of the primary mirror supporting structure is of crucial importance. Now the general method is to use ANSYS APDL programming, which is inconvenient and complex to fit for the different components, the calculation require much time and the analysis is lack of efficient. Taking the diameter of 1.12 m telescope primary mirror as the research objection, the paper combine the actual design parameters of SONG telescope, respectively using ANSYS WORKBENCH to employ the primary mirror axial and lateral support model in finite element method, the optimal solution is obtained by optimization design and the change rule of mirror surface deformation under inclined condition is studied. The optimization results according with the requirements of the primary mirror comprehensive error proves that the optimization analysis method is available and applicable.

  13. Development of large-aperture electro-optical switch for high power laser at CAEP

    NASA Astrophysics Data System (ADS)

    Zhang, Xiongjun; Wu, Dengsheng; Zhang, Jun; Lin, Donghui; Zheng, Jiangang; Zheng, Kuixing

    2015-02-01

    Large-aperture electro-optical switch based on plasma Pockels cell (PPC) is one of important components for inertial confinement fusion (ICF) laser facility. We have demonstrated a single-pulse driven 4×1 PPC with 400mm×400mm aperture for SGIII laser facility. And four 2×1 PPCs modules with 350mm×350mm aperture have been operated in SGII update laser facility. It is different to the PPC of NIF and LMJ for its simple operation to perform Pockels effect. With optimized operation parameters, the PPCs meet the SGII-U laser requirement of four-pass amplification control. Only driven by one high voltage pulser, the simplified PPC system would be provided with less associated diagnostics, and higher reliability. To farther reduce the insert loss of the PPC, research on the large-aperture PPC based on DKDP crystal driven by one pulse is developed. And several single-pulse driven PPCs with 80mm×80mm DKDP crystal have been manufactured and operated in laser facilities.

  14. Recent Enhancements of the Phased Array Mirror Extendible Large Aperture (PAMELA) Telescope Testbed at MSFC

    NASA Technical Reports Server (NTRS)

    Rakoczy, John; Burdine, Robert (Technical Monitor)

    2001-01-01

    Recent incremental upgrades to the Phased Array Mirror Extendible Large Aperture (PAMELA) telescope testbed have enabled the demonstration of phasing (with a monochromatic source) of clusters of primary mirror segments down to the diffraction limit. PAMELA upgrades include in improved Shack-Hartmann wavefront sensor, passive viscoelastic damping treatments for the voice-coil actuators, mechanical improvement of mirror surface figures, and optical bench baffling. This report summarizes the recent PAMELA upgrades, discusses the lessons learned, and presents a status of this unique testbed for wavefront sensing and control. The Marshall Space Flight Center acquired the Phased Array Mirror Extendible Large Aperture (PAMELA) telescope in 1993 after Kaman Aerospace was unable to complete integration and testing under the limited SDIO and DARPA funding. The PAMELA is a 36-segment, half-meter aperture, adaptive telescope which utilizes a Shack-Hartmann wavefront sensor, inductive coil edge sensors, voice coil actuators, imaging CCD cameras and interferometry for figure alignment, wavefront sensing and control. MSFC originally obtained the PAMELA to supplement its research in the interactions of control systems with flexible structures. In August 1994, complete tip, tilt and piston control was successfully demonstrated using the Shack-Hartmann wavefront sensor and the inductive edge sensors.

  15. Research on the support structure of the primary mirror of large-aperture telescope

    NASA Astrophysics Data System (ADS)

    Wang, Yang; Zhang, Jingxu

    2007-12-01

    Large-aperture telescope can be used in surveying battlefield, researching landform, searching object, real-time monitoring, imaging, detecting and identifying spatial targets and so on. A large-aperture telescope for achieving high resolution power is designed to monitor spatial target and image in real time. Real-time monitoring plays an important role in military conflicts. The orbit parameter of object, quantity, geometrical shape parameter and so on can be obtained by detect spatial target. With the development of optical technology, people require larger aperture in optics-electronic (O-E) system. By increasing optical aperture, the ability of collecting light and resolution power in the system can be enhanced. But the support structure of the primary mirror of large-aperture telescope will be a very difficult problem. With the increase of primary mirror aperture, the weight of the primary mirror will become larger than before. The root mean square (rms) of the primary mirror is affected by many factors, such as deadweight, deformation of heat, environment and so on. Due to the primary mirror of telescope is an important component of telescope system. By reducing the weight of primary mirror, precision of the system is ensured. During the designing phase, one can consider the supporting project of the primary mirror synthetically and analyze it roundly according to technical requirement of optical system and the effect factors. The final structural design can be reasonable. In an astronomical telescope, the surface of reflector is an important part for collecting dark radiation of celestial bodies. Its surface shape will have an effect on collecting efficiency of telescope radiant energy directly. So the rms must be very high. Optical system of large aperture, small wavelength and small focus can receive maximal light intensity. For ground-based optical astronomical telescope, the design proposed in the paper can satisfy the requirement of the possible

  16. How Membrane-Active Peptides Get into Lipid Membranes.

    PubMed

    Sani, Marc-Antoine; Separovic, Frances

    2016-06-21

    The structure-function relationship for a family of antimicrobial peptides (AMPs) from the skin of Australian tree frogs is discussed and compared with that of peptide toxins from bee and Australian scorpion venoms. Although these membrane-active peptides induce a similar cellular fate by disrupting the lipid bilayer integrity, their lytic activity is achieved via different modes of action, which are investigated in relation to amino acid sequence, secondary structure, and membrane lipid composition. In order to better understand what structural features govern the interaction between peptides and lipid membranes, cell-penetrating peptides (CPPs), which translocate through the membrane without compromising its integrity, are also discussed. AMPs possess membrane lytic activities that are naturally designed to target the cellular membrane of pathogens or competitors. They are extremely diverse in amino acid composition and often show specificity against a particular strain of microbe. Since our antibiotic arsenal is declining precariously in the face of the rise in multiantibiotic resistance, AMPs increasingly are seen as a promising alternative. In an effort to understand their molecular mechanism, biophysical studies of a myriad of AMPs have been reported, yet no unifying mechanism has emerged, rendering difficult the rational design of drug leads. Similarly, a wide variety of cytotoxic peptides are found in venoms, the best known being melittin, yet again, predicting their activity based on a particular amino acid composition or secondary structure remains elusive. A common feature of these membrane-active peptides is their preference for the lipid environment. Indeed, they are mainly unstructured in solution and, in the presence of lipid membranes, quickly adsorb onto the surface, change their secondary structure, eventually insert into the hydrophobic core of the membrane bilayer, and finally disrupt the bilayer integrity. These steps define the molecular

  17. End-to-End Assessment of a Large Aperture Segmented Ultraviolet Optical Infrared (UVOIR) Telescope Architecture

    NASA Technical Reports Server (NTRS)

    Feinberg, Lee; Rioux, Norman; Bolcar, Matthew; Liu, Alice; Guyon, Oliver; Stark, Chris; Arenberg, Jon

    2016-01-01

    Key challenges of a future large aperture, segmented Ultraviolet Optical Infrared (UVOIR) Telescope capable of performing a spectroscopic survey of hundreds of Exoplanets will be sufficient stability to achieve 10^-10 contrast measurements and sufficient throughput and sensitivity for high yield Exo-Earth spectroscopic detection. Our team has collectively assessed an optimized end to end architecture including a high throughput coronagraph capable of working with a segmented telescope, a cost-effective and heritage based stable segmented telescope, a control architecture that minimizes the amount of new technologies, and an Exo-Earth yield assessment to evaluate potential performance. These efforts are combined through integrated modeling, coronagraph evaluations, and Exo-Earth yield calculations to assess the potential performance of the selected architecture. In addition, we discusses the scalability of this architecture to larger apertures and the technological tall poles to enabling it.

  18. Advanced Technology Large-Aperture Space Telescope: Science Drivers and Technology Developments

    NASA Technical Reports Server (NTRS)

    Postman, Marc; Brown, Tom; Sembach, Kenneth; Glavallsco, Mauro; Traub, Wesley; Stapelfeldt, Karl; Calzetti, Daniela; Oegerle, William; Rich, R. Michael; Stahl, H. Philip; Tumlinson, Jason; Mountain, Matt; Soummer, Remi; Hyde, Tupper

    2012-01-01

    The Advanced Technology Large-Aperture Space Telescope (ATLAST) is a concept for an 8- to 16-m ultraviolet optical near Infrared space observatory for launch in the 2025 to 2030 era. ATLAST will allow astronomers to answer fundamental questions at the forefront of modern astrophysics, including: Is there life elsewhere in the Galaxy? We present a range of science drivers and the resulting performance requirements for ATLAST (8- to 16-marcsec angular resolution, diffraction limited imaging at 0.5 micron wavelength, minimum collecting area of 45 sq m, high sensitivity to light wavelengths from 0.1 to 2.4 micron, high stability in wavefront sensing and control). We also discuss the priorities for technology development needed to enable the construction of ATLAST for a cost that is comparable to that of current generation observatory-class space missions.

  19. Simulation analysis of on-orbit adjustment and compensation for large aperture optical system

    NASA Astrophysics Data System (ADS)

    Liu, Jianfeng; Li, Bo; Sun, Dewei; Ruan, Ningjuan; Zhou, Feng

    2014-09-01

    With the uprating requirements of space remote sensing, the aperture of the optical remote sensor is getting larger and larger. The influences of both the support of optical elements and gravity deformation on the optical system are difficult to conquer. Therefore, it is necessary to compensate the descending optical performance which is caused by the surface error of primary mirror by means of adjusting the position parameters of the optical elements on-orbit. A large aperture coaxial three-mirror optical system is introduced in the paper. Matlab and MetroPro are used to simulate the surface error of the primary mirror. The surface error of the primary mirror is compensated by adjusting the position freedoms of the secondary mirror. The results show that the adjustment of the position freedoms of the secondary mirror can compensate both the coma and some astigmatism of the primary mirror, but not the spherical aberration.

  20. A Large Aperture, High Energy Laser System for Optics and Optical Component Testing

    SciTech Connect

    Nostrand, M C; Weiland, T L; Luthi, R L; Vickers, J L; Sell, W D; Stanley, J A; Honig, J; Auerbach, J; Hackel, R P; Wegner, P J

    2003-11-01

    A large aperture, kJ-class, multi-wavelength Nd-glass laser system has been constructed at Lawrence Livermore National Lab which has unique capabilities for studying a wide variety of optical phenomena. The master-oscillator, power-amplifier (MOPA) configuration of this ''Optical Sciences Laser'' (OSL) produces 1053 nm radiation with shaped pulse lengths which are variable from 0.1-100 ns. The output can be frequency doubled or tripled with high conversion efficiency with a resultant 100 cm{sup 2} high quality output beam. This facility can accommodate prototype hardware for large-scale inertial confinement fusion lasers allowing for investigation of integrated system issues such as optical lifetime at high fluence, optics contamination, compatibility of non-optical materials, and laser diagnostics.

  1. ATLAST-9.2m: a Large-Aperture Deployable Space Telescope

    NASA Technical Reports Server (NTRS)

    Oergerle, William; Feinberg, Lee D.; Purves, Lloyd R.; Hyde, T. Tupper; Thronson, Harley A.; Townsend, Jacqueline A.; Postman, Marc; Bolear, Matthew R.; Budinoff, Jason G.; Dean, Bruce H.; Clampin, Mark C.; Ebbets, Dennis C.; Gong, Qian; Gull, Theodore R.; Howard, Joseph M.; Jones, Andrew L.; Lyon, Richard G.; Pasquale, Bert A.; Perrygo, Charles; Smith, Jeffrey S.; Thompson, Patrick L.; Woodgate, Bruce E.

    2010-01-01

    We present results of a study of a deployable version of the Advanced Technology Large-Aperture Space Telescope (ATLAST), designed to operate in a Sun-Earth L2 orbit. The primary mirror of the segmented 9.2-meter aperture has 36 hexagonal 1.315 m (flat to flat) glass mirrors. The architecture and folding of the telescope is similar to JWST, allowing it to fit into the 6.5 m fairing of a modest upgrade to the Delta-IV Heavy version of the Evolved Expendable Launch Vehicle (EELV). We discuss the overall observatory design, optical design, instruments, stray light, wavefront sensing and control, pointing and thermal control, and in-space servicing options.

  2. Data correction techniques for the airborne large-aperture static image spectrometer based on image registration

    NASA Astrophysics Data System (ADS)

    Zhang, Geng; Shi, Dalian; Wang, Shuang; Yu, Tao; Hu, Bingliang

    2015-01-01

    We propose an approach to correct the data of the airborne large-aperture static image spectrometer (LASIS). LASIS is a kind of stationary interferometer which compromises flux output and device stability. It acquires a series of interferograms to reconstruct the hyperspectral image cube. Reconstruction precision of the airborne LASIS data suffers from the instability of the plane platform. Usually, changes of plane attitudes, such as yaws, pitches, and rolls, can be precisely measured by the inertial measurement unit. However, the along-track and across-track translation errors are difficult to measure precisely. To solve this problem, we propose a co-optimization approach to compute the translation errors between the interferograms using an image registration technique which helps to correct the interferograms with subpixel precision. To demonstrate the effectiveness of our approach, experiments are run on real airborne LASIS data and our results are compared with those of the state-of-the-art approaches.

  3. A conceptual design of a large aperture microwave radiometer geostationary platform

    NASA Technical Reports Server (NTRS)

    Garn, Paul A.; Garrison, James L.; Jasinski, Rachel

    1992-01-01

    A conceptual design of a Large Aperture Microwave Radiometer (LAMR) Platform has been developed and technology areas essential to the design and on-orbit viability of the platform have been defined. Those technologies that must be developed to the requirement stated here for the LAMR mission to be viable include: advanced radiation resistant solar cells, integrated complex structures, large segmented reflector panels, sub 3 kg/m(exp 2) areal density large antennas, and electric propulsion systems. Technology areas that require further development to enhance the capabilities of the LAMR platform (but are not essential for viability) include: electrical power storage, on-orbit assembly, and on-orbit systems checkout and correction.

  4. Fluid transport by active elastic membranes

    NASA Astrophysics Data System (ADS)

    Evans, Arthur A.; Lauga, Eric

    2011-09-01

    A flexible membrane deforming its shape in time can self-propel in a viscous fluid. Alternatively, if the membrane is anchored, its deformation will lead to fluid transport. Past work in this area focused on situations where the deformation kinematics of the membrane were prescribed. Here we consider models where the deformation of the membrane is not prescribed, but instead the membrane is internally forced. Both the time-varying membrane shape and the resulting fluid motion result then from a balance between prescribed internal active stresses, internal passive resistance, and external viscous stresses. We introduce two specific models for such active internal forcing: one where a distribution of active bending moments is prescribed, and one where active inclusions exert normal stresses on the membrane by pumping fluid through it. In each case, we asymptotically calculate the membrane shape and the fluid transport velocities for small forcing amplitudes, and recover our results using scaling analysis.

  5. Pulse power requirements for large aperture optical switches based on plasma electrode Pockels cells

    SciTech Connect

    Rhodes, M.A.; Taylor, J.

    1992-06-01

    We discuss very large-aperture optical switches (greater than 30 [times] 30 cm) as an enabling technology for inertial confinement fusion drivers based on multipass laser amplifiers. Large-scale laser fusion drivers such as the Nova laser have been based on single-pass amplifier designs in part because of the unavailability of a suitable large-aperture switch. We are developing an optical switch based on a Pockels cell employing plasma-electrodes. A plasma-electrode Pockels cell (PEPC) is a longitudinal-mode Pockels cell in which a plasma discharge is formed on each side of an electro-optic crystal (typically KDP or deuterated KDP, often designated KD*P). The plasmas formed on either side of the crystal act as transparent electrodes for a switching-pulse and are intended to allow uniform charging of the entire crystal. The switching-pulse is a nominally rectangular high-voltage pulse equal to the half-wave voltage V[sub x] ( 8 kV for KD*P or 17 kV for KDP) and is applied across the crystal via the plasma-electrodes. When the crystal is charged to V[sub x], the polarization of an incoming, linearly polarized, laser beam is rotated by 90[degree]. When used in conjunction with an appropriate, passive polarizer, an optical switch is thus realized. A switch with a clear aperture of 37 [times] 37 cm is now in construction for the Beamlet laser which will serve as a test bed for this switch as well as other technologies required for an advanced NOVA laser design. In this paper, we discuss the unique power electronics requirements of PEPC optical switches.

  6. Pulse power requirements for large aperture optical switches based on plasma electrode Pockels cells

    SciTech Connect

    Rhodes, M.A.; Taylor, J.

    1992-06-01

    We discuss very large-aperture optical switches (greater than 30 {times} 30 cm) as an enabling technology for inertial confinement fusion drivers based on multipass laser amplifiers. Large-scale laser fusion drivers such as the Nova laser have been based on single-pass amplifier designs in part because of the unavailability of a suitable large-aperture switch. We are developing an optical switch based on a Pockels cell employing plasma-electrodes. A plasma-electrode Pockels cell (PEPC) is a longitudinal-mode Pockels cell in which a plasma discharge is formed on each side of an electro-optic crystal (typically KDP or deuterated KDP, often designated KD*P). The plasmas formed on either side of the crystal act as transparent electrodes for a switching-pulse and are intended to allow uniform charging of the entire crystal. The switching-pulse is a nominally rectangular high-voltage pulse equal to the half-wave voltage V{sub x} ( 8 kV for KD*P or 17 kV for KDP) and is applied across the crystal via the plasma-electrodes. When the crystal is charged to V{sub x}, the polarization of an incoming, linearly polarized, laser beam is rotated by 90{degree}. When used in conjunction with an appropriate, passive polarizer, an optical switch is thus realized. A switch with a clear aperture of 37 {times} 37 cm is now in construction for the Beamlet laser which will serve as a test bed for this switch as well as other technologies required for an advanced NOVA laser design. In this paper, we discuss the unique power electronics requirements of PEPC optical switches.

  7. Large Aperture "Photon Bucket" Optical Receiver Performance in High Background Environments

    NASA Technical Reports Server (NTRS)

    Vilnrotter, Victor A.; Hoppe, D.

    2011-01-01

    The potential development of large aperture groundbased "photon bucket" optical receivers for deep space communications, with acceptable performance even when pointing close to the sun, is receiving considerable attention. Sunlight scattered by the atmosphere becomes significant at micron wavelengths when pointing to a few degrees from the sun, even with the narrowest bandwidth optical filters. In addition, high quality optical apertures in the 10-30 meter range are costly and difficult to build with accurate surfaces to ensure narrow fields-of-view (FOV). One approach currently under consideration is to polish the aluminum reflector panels of large 34-meter microwave antennas to high reflectance, and accept the relatively large FOV generated by state-of-the-art polished aluminum panels with rms surface accuracies on the order of a few microns, corresponding to several-hundred micro-radian FOV, hence generating centimeter-diameter focused spots at the Cassegrain focus of 34-meter antennas. Assuming pulse-position modulation (PPM) and Poisson-distributed photon-counting detection, a "polished panel" photon-bucket receiver with large FOV will collect hundreds of background photons per PPM slot, along with comparable signal photons due to its large aperture. It is demonstrated that communications performance in terms of PPM symbol-error probability in high-background high-signal environments depends more strongly on signal than on background photons, implying that large increases in background energy can be compensated by a disproportionally small increase in signal energy. This surprising result suggests that large optical apertures with relatively poor surface quality may nevertheless provide acceptable performance for deep-space optical communications, potentially enabling the construction of cost-effective hybrid RF/optical receivers in the future.

  8. Topology optimization-based lightweight primary mirror design of a large-aperture space telescope.

    PubMed

    Liu, Shutian; Hu, Rui; Li, Quhao; Zhou, Ping; Dong, Zhigang; Kang, Renke

    2014-12-10

    For the large-aperture space telescope, the lightweight primary mirror design with a high-quality optical surface is a critical and challenging issue. This work presents a topology optimization-based design procedure for a lightweight primary mirror and a new mirror configuration of a large-aperture space telescope is obtained through the presented design procedure. Inspired by the topology optimization method considering cast constraints, an optimization model for the configuration design of the mirror back is proposed, through which the distribution and the heights of the stiffeners on the mirror back can be optimized simultaneously. For the purpose of minimizing the optical surface deviation due to self-weight and polishing pressure loadings, the objective function is selected as to maximize the mirror structural stiffness, which can be achieved by minimizing the structural compliance. The total mass of the primary mirror is assigned as the constraint. In the application example, results of the optimized design topology for two kinds of mass constraints are presented. Executing the design procedure for specific requirements and postprocessing the topology obtained of the structure, a new mirror configuration with tree-like stiffeners and a multiple-arch back in double directions is proposed. A verification model is constructed to evaluate the design results and the finite element method is used to calculate the displacement of the mirror surface. Then the RMS deviation can be obtained after fitting the deformed surface by Zernike polynomials. The proposed mirror is compared with two classical mirrors in the optical performance, and the comparison results demonstrate the superiority of the new mirror configuration. PMID:25608076

  9. Large-aperture prism-array lens for high-energy X-ray focusing.

    PubMed

    Zhang, Weiwei; Liu, Jing; Chang, Guangcai; Shi, Zhan; Li, Ming; Ren, Yuqi; Zhang, Xiaowei; Yi, Futing; Liu, Peng; Sheng, Weifan

    2016-09-01

    A new prism-array lens for high-energy X-ray focusing has been constructed using an array of different prisms obtained from different parabolic structures by removal of passive parts of material leading to a multiple of 2π phase variation. Under the thin-lens approximation the phase changes caused by this lens for a plane wave are exactly the same as those caused by a parabolic lens without any additional corrections when they have the same focal length, which will provide good focusing; at the same time, the total transmission and effective aperture of this lens are both larger than those of a compound kinoform lens with the same focal length, geometrical aperture and feature size. This geometry can have a large aperture that is not limited by the feature size of the lens. Prototype nickel lenses with an aperture of 1.77 mm and focal length of 3 m were fabricated by LIGA technology, and were tested using CCD camera and knife-edge scan method at the X-ray Imaging and Biomedical Application Beamline BL13W1 at Shanghai Synchrotron Radiation Facility, and provided a focal width of 7.7 µm and a photon flux gain of 14 at an X-ray energy of 50 keV. PMID:27577761

  10. Design of large aperture superferric quadrupole magnets for an in-flight fragment separator

    SciTech Connect

    Zaghloul, Aziz; Kim, Dogyun; Kim, Jangyoul; Kim, Mijung; Kim, Myeongjin; Yun, Chongcheoul; Kim, Jongwon

    2014-01-29

    Superferric quadrupole magnets to be used for in-flight fragment separator have been designed. A quadrupole magnet triplet for beam focusing is placed in a cryostat together with superconducting correction coils. To maximize acceptance of rare isotope beams produced by projectile fragmentation, it is essential to use large-aperture quadrupole magnets. The pole tip radius is 17 cm in the current design, and we tried to enlarge the aperture with 3D analysis on magnetic fields. In the front end of the separator, where a target and beam dump are located, we plan to use two sets of quadrupole triplets made of high-Tc superconductor (HTS) operating at 20-50 K considering high radiation heat load. The HTS magnet will use warm iron poles. Both low-Tc and high-Tc superconductors are acquired for test winding, and two kinds of dewar and cryostat are under construction to perform the coil and magnet tests. The magnetic design of superferric quadrupole is mainly discussed.

  11. Large-aperture wide-bandwidth antireflection-coated silicon lenses for millimeter wavelengths.

    PubMed

    Datta, R; Munson, C D; Niemack, M D; McMahon, J J; Britton, J; Wollack, E J; Beall, J; Devlin, M J; Fowler, J; Gallardo, P; Hubmayr, J; Irwin, K; Newburgh, L; Nibarger, J P; Page, L; Quijada, M A; Schmitt, B L; Staggs, S T; Thornton, R; Zhang, L

    2013-12-20

    The increasing scale of cryogenic detector arrays for submillimeter and millimeter wavelength astrophysics has led to the need for large aperture, high index of refraction, low loss, cryogenic refracting optics. Silicon with n=3.4, low loss, and high thermal conductivity is a nearly optimal material for these purposes but requires an antireflection (AR) coating with broad bandwidth, low loss, low reflectance, and a matched coefficient of thermal expansion. We present an AR coating for curved silicon optics comprised of subwavelength features cut into the lens surface with a custom three-axis silicon dicing saw. These features constitute a metamaterial that behaves as a simple dielectric coating. We have fabricated silicon lenses as large as 33.4 cm in diameter with micromachined layers optimized for use between 125 and 165 GHz. Our design reduces average reflections to a few tenths of a percent for angles of incidence up to 30° with low cross polarization. We describe the design, tolerance, manufacture, and measurements of these coatings and present measurements of the optical properties of silicon at millimeter wavelengths at cryogenic and room temperatures. This coating and lens fabrication approach is applicable from centimeter to submillimeter wavelengths and can be used to fabricate coatings with greater than octave bandwidth. PMID:24513939

  12. The measurement and analysis of wavefront structure from large aperture ICF optics

    SciTech Connect

    Wolfe, C.R.; Lawson, J.K.

    1995-05-30

    This paper discusses the techniques, developed over the past year, for high spatial resolution measurement and analysis of the transmitted and/or reflected wavefront of large aperture ICF optical components. Parts up to 400 mm {times} 750 mm have been measured and include: laser slabs, windows, KDP crystals and lenses. The measurements were performed using state-of-the-art commercial phase shifting interferometers at a wavelength of 633 {mu}m. Both 1 and 2-D Fourier analysis have been used to characterize the wavefront; specifically the Power Spectral Density, (PSD), function was calculated. The PSDs of several precision optical components will be shown. The PSD(V) is proportional to the (amplitude){sup 2} of components of the Fourier frequency spectrum. The PSD describes the scattered intensity and direction as a function of scattering angle in the wavefront. The capability of commercial software is limited to 1-D Fourier analysis only. We are developing our own 2-D analysis capability in support of work to revise specifications for NIF optics. 2-D analysis uses the entire wavefront phase map to construct 2D PSD functions. We have been able to increase the signal-to-noise relative to 1-D and can observe very subtle wavefront structure.

  13. A Future Large-Aperture UVOIR Space Observatory: Key Technologies and Capabilities

    NASA Technical Reports Server (NTRS)

    Bolcar, Matthew Ryan; Stahle, Carl M.; Balasubramaniam, Kunjithapatham; Clampin, Mark; Feinberg, Lee D.; Mosier, Gary E.; Quijada, Manuel A.; Rauscher, Bernard J.; Redding, David C.; Rioux, Norman M.; Shaklan, Stuart B.; Stahl, H. Philip; Thronson, Harley A.

    2015-01-01

    We present the key technologies and capabilities that will enable a future, large-aperture ultravioletopticalinfrared (UVOIR) space observatory. These include starlight suppression systems, vibration isolation and control systems, lightweight mirror segments, detector systems, and mirror coatings. These capabilities will provide major advances over current and near-future observatories for sensitivity, angular resolution, and starlight suppression. The goals adopted in our study for the starlight suppression system are 10-10 contrast with an inner working angle of 20 milliarcsec and broad bandpass. We estimate that a vibration and isolation control system that achieves a total system vibration isolation of 140 dB for a vibration-isolated mass of 5000 kg is required to achieve the high wavefront error stability needed for exoplanet coronagraphy. Technology challenges for lightweight mirror segments include diffraction-limited optical quality and high wavefront error stability as well as low cost, low mass, and rapid fabrication. Key challenges for the detector systems include visible-blind, high quantum efficiency UV arrays, photon counting visible and NIR arrays for coronagraphic spectroscopy and starlight wavefront sensing and control, and detectors with deep full wells with low persistence and radiation tolerance to enable transit imaging and spectroscopy at all wavelengths. Finally, mirror coatings with high reflectivity ( 90), high uniformity ( 1) and low polarization ( 1) that are scalable to large diameter mirror substrates will be essential for ensuring that both high throughput UV observations and high contrast observations can be performed by the same observatory.

  14. Large-Aperture Wide-Bandwidth Anti-Reflection-Coated Silicon Lenses for Millimeter Wavelengths

    NASA Technical Reports Server (NTRS)

    Datta, R.; Munson, C. D.; Niemack, M. D.; McMahon, J. J.; Britton, J.; Wollack, E. J.; Beall, J.; Devlin, M. J.; Fowler, J.; Gallardo, P.; Hubmayr, J.; Irwin, K.; Newburgh, L.; Nibarger, J. P.; Page, L.; Quijada, M. A.; Schmitt, B. L.; Staggs, S. T.; Thornton, R.; Zhang, L.

    2013-01-01

    The increasing scale of cryogenic detector arrays for sub-millimeter and millimeter wavelength astrophysics has led to the need for large aperture, high index of refraction, low loss, cryogenic refracting optics. Silicon with n = 3.4, low loss, and relatively high thermal conductivity is a nearly optimal material for these purposes, but requires an antireflection (AR) coating with broad bandwidth, low loss, low reflectance, and a matched coffecient of thermal expansion. We present an AR coating for curved silicon optics comprised of subwavelength features cut into the lens surface with a custom three axis silicon dicing saw. These features constitute a metamaterial that behaves as a simple dielectric coating. We have fabricated and coated silicon lenses as large as 33.4 cm in diameter with coatings optimized for use between 125-165 GHz. Our design reduces average reflections to a few tenths of a percent for angles of incidence up to 30 deg. with low cross-polarization. We describe the design, tolerance, manufacture, and measurements of these coatings and present measurements of the optical properties of silicon at millimeter wavelengths at cryogenic and room temperatures. This coating and lens fabrication approach is applicable from centimeter to sub-millimeter wavelengths and can be used to fabricate coatings with greater than octave bandwidth.

  15. Large-aperture Wide-bandwidth Antireflection-coated Silicon Lenses for Millimeter Wavelengths

    NASA Technical Reports Server (NTRS)

    Datta, R.; Munson, C. D.; Niemack, M. D.; McMahon, J. J.; Britton, J.; Wollack, Edward J.; Beall, J.; Devlin, M. J.; Fowler, J.; Gallardo, P.; Hubmayr, J.; Irwin, K.; Newburgh, L.; Nibarger, J. P.; Page, L.; Quijada, Manuel A.; Schmitt, B. L.; Staggs, S. T.; Thornton, R.; Zhang, L.

    2013-01-01

    The increasing scale of cryogenic detector arrays for submillimeter and millimeter wavelength astrophysics has led to the need for large aperture, high index of refraction, low loss, cryogenic refracting optics. Silicon with n 3.4, low loss, and high thermal conductivity is a nearly optimal material for these purposes but requires an antireflection (AR) coating with broad bandwidth, low loss, low reflectance, and a matched coefficient of thermal expansion. We present an AR coating for curved silicon optics comprised of subwavelength features cut into the lens surface with a custom three-axis silicon dicing saw. These features constitute a metamaterial that behaves as a simple dielectric coating.We have fabricated silicon lenses as large as 33.4 cm in diameter with micromachined layers optimized for use between 125 and 165 GHz. Our design reduces average reflections to a few tenths of a percent for angles of incidence up to 30deg with low cross polarization.We describe the design, tolerance, manufacture, and measurements of these coatings and present measurements of the optical properties of silicon at millimeter wavelengths at cryogenic and room temperatures. This coating and lens fabrication approach is applicable from centimeter to submillimeter wavelengths and can be used to fabricate coatings with greater than octave bandwidth.

  16. [Modeling and simulation of effect of optical distortion on the large aperture static imaging spectrometer].

    PubMed

    Lü, Qun-bo; Xiangli, Bin; Yao, Tao; Jing, Juan-juan; Wang, Zhong-hou

    2010-01-01

    As a new type Fourier transform imaging spectrometry, large aperture static imaging spectrometry (LASIS) has come forth in recent years, which has many advantages such as simple principle, high stability and so on. However, the requirement for the optical system design of LASIS was very harsh. As one of the optical aberrations, optical distortion degrades the data quality acquired by LASIS, consequently limits its applications. According to the analysis of the data acquisition principle of LASIS, the data model with the effect of optical distortion was presented, which could be used for LASIS performance pre-evaluation. Finally, the computer simlulation of the data model was achieved with supposed parameters. The simulation results indicated that the relative error more than 5% was induced in the recovery spectrum, and approximate 8 nm spectral line deviation was occurred at the long wavelength region. The results show that the 4% optical distortion was inapplicable for LASIS although it is acceptant for common optical imaging system. PMID:20302101

  17. High resolution beamforming on large aperture vertical line arrays: Processing synthetic data

    NASA Astrophysics Data System (ADS)

    Tran, Jean-Marie Q.; Hodgkiss, William S.

    1990-09-01

    This technical memorandum studies the beamforming of large aperture line arrays deployed vertically in the water column. The work concentrates on the use of high resolution techniques. Two processing strategies are envisioned: (1) full aperture coherent processing which offers in theory the best processing gain; and (2) subaperture processing which consists in extracting subapertures from the array and recombining the angular spectra estimated from these subarrays. The conventional beamformer, the minimum variance distortionless response (MVDR) processor, the multiple signal classification (MUSIC) algorithm and the minimum norm method are used in this study. To validate the various processing techniques, the ATLAS normal mode program is used to generate synthetic data which constitute a realistic signals environment. A deep-water, range-independent sound velocity profile environment, characteristic of the North-East Pacific, is being studied for two different 128 sensor arrays: a very long one cut for 30 Hz and operating at 20 Hz; and a shorter one cut for 107 Hz and operating at 100 Hz. The simulated sound source is 5 m deep. The full aperture and subaperture processing are being implemented with curved and plane wavefront replica vectors. The beamforming results are examined and compared to the ray-theory results produced by the generic sonar model.

  18. Improving 351-nm Damage Performance of Large-Aperture Fused Silica and DKDP Optics

    SciTech Connect

    Burnham, A K; Hackel, L; Wegner, P; Parham, T; Hrubesh, L; Penetrante, B; Whitman, P; Demos, S; Menapace, J; Runkel, M; Fluss, M; Feit, M; Key, M; Biesiada, T

    2002-01-07

    A program to identify and eliminate the causes of UV laser-induced damage and growth in fused silica and DKDP has developed methods to extend optics lifetimes for large-aperture, high-peak-power, UV lasers such as the National Ignition Facility (NIF). Issues included polish-related surface damage initiation and growth on fused silica and DKDP, bulk inclusions in fused silica, pinpoint bulk damage in DKDP, and UV-induced surface degradation in fused silica and DKDP in a vacuum. Approaches included an understanding of the mechanism of the damage, incremental improvements to existing fabrication technology, and feasibility studies of non-traditional fabrication technologies. Status and success of these various approaches are reviewed. Improvements were made in reducing surface damage initiation and eliminating growth for fused silica by improved polishing and post-processing steps, and improved analytical techniques are providing insights into mechanisms of DKDP damage. The NIF final optics hardware has been designed to enable easy retrieval, surface-damage mitigation, and recycling of optics.

  19. Large-aperture broadband sapphire windows for common aperture, target acquisition, tracking, and surveillance systems

    NASA Astrophysics Data System (ADS)

    Askinazi, Joel

    1997-06-01

    State of the art optical sensing systems performing target acquisition/tracking and surveillance functions are being designed to incorporate a number of sensors into one package. These include visual and MWIR cameras, FLIRs, and laser range finders. These combined systems are being configured to view through a common aperture window. Typical window diameters are to eleven inches, but some surveillance applications have windows approaching twenty inches in diameter. These sensor windows typically operate in hostile environments including very high pressure differentials, large thermal gradients, and severe rain and sand abrasion. EMI/EMC protection and de-icing capabilities are also commonly required. For airborne applications and to minimize thermal gradients, thinner, lightweight, high strength windows are also necessary. Sapphire is an ideal window material to satisfy these requirements due to its high strength, UV-MWIR bandpass, minimal optical scatter, excellent index of refraction homogeneity and very high scratch/impact resistance. Associated optical fabrication, grid lithography and optical coating processes have been developed at Hughes Danbury for sapphire windows. This paper addresses the development of a family of large aperture, broadband sapphire windows which also provide EMI/EMC protection and de-icing capabilities. The resulting design configuration and performance characteristics are also addressed. Future technology development requirements are also discussed.

  20. 8 Meter Advanced Technology Large-Aperture Space Telescope (ATLAST-8m)

    NASA Technical Reports Server (NTRS)

    Stahl, H. Philip

    2010-01-01

    ATLAST-8m (Advanced Technology Large Aperture Space Telescope) is a proposed 8-meter monolithic UV/optical/NIR space observatory (wavelength range 110 to 2500 nm) to be placed in orbit at Sun-Earth L2 by NASA's planned Ares V heavy lift vehicle. Given its very high angular resolution (15 mas @ 500 nm), sensitivity and performance stability, ATLAST-8m is capable of achieving breakthroughs in a broad range of astrophysics including: Is there life elsewhere in the Galaxy? An 8-meter UVOIR observatory has the performance required to detect habitability (H2O, atmospheric column density) and biosignatures (O2, O3, CH4) in terrestrial exoplanet atmospheres, to reveal the underlying physics that drives star formation, and to trace the complex interactions between dark matter, galaxies, and intergalactic medium. The ATLAST Astrophysics Strategic Mission Concept Study developed a detailed point design for an 8-m monolithic observatory including optical design; structural design/analysis including primary mirror support structure, sun shade and secondary mirror support structure; thermal analysis; spacecraft including structure, propulsion, GN&C, avionics, power systems and reaction wheels; mass and power budgets; and system cost. The results of which were submitted by invitation to NRC's 2010 Astronomy & Astrophysics Decadal Survey.

  1. Origins of high-frequency scattered waves near PKKP from large aperture seismic array data

    USGS Publications Warehouse

    Earle, P.S.

    2002-01-01

    This article identifies the likely origin of 1-Hz scattered waves in the vicinity of PKKP by comparing measurements of slowness and onset time to ray-theoretical predictions. The measurements are obtained from slant stacks of Large Aperture Seismic Array (LASA) data from 36 earthquakes and six explosions in the range 30??-116??. Three types of scattered waves explain the main features seen in the stacks, including: P scattered to PKP near the Earth's surface (P.PKP), PKKP scattered near its core-mantle-boundary (CMB) reflection point (PK.KP), and SKKP scattered near its CMB reflection point (SK.KP). The LASA stacks image the amplitude and slowness variations of the scattered waves with time. They also show where these waves can be detected and where they are free from contaminating arrivals. SK.KP waves rise above the noise approximately 100 sec before the onset time of the main SKKP arrival near 113??. Observations of PK.KP span 30??-100??. However, at distances greater than 50?? they suffer from P.PKP contamination. At distances less than 40?? the PK.KP last for about 280 sec. This is approximately 130 sec longer than the maximum ray-theoretical prediction for waves scattered at the CMB, indicating a possible combination of near-surface scattering and contributions from the overlying mantle.

  2. Intense THz Pulses with large ponderomotive potential generated from large aperture photoconductive antennas.

    PubMed

    Ropagnol, X; Khorasaninejad, M; Raeiszadeh, M; Safavi-Naeini, S; Bouvier, M; Côté, C Y; Laramée, A; Reid, M; Gauthier, M A; Ozaki, T

    2016-05-30

    We report the generation of free space terahertz (THz) pulses with energy up to 8.3 ± 0.2 µJ from an encapsulated interdigitated ZnSe Large Aperture Photo-Conductive Antenna (LAPCA). An aperture of 12.2 cm2 is illuminated using a 400 nm pump laser with multi-mJ energies at 10 Hz repetition rate. The calculated THz peak electric field is 331 ± 4 kV/cm with a spectrum characterized by a median frequency of 0.28 THz. Given its relatively low frequency, this THz field will accelerate charged particles efficiently having very large ponderomotive energy of 15 ± 1 eV for electrons in vacuum. The scaling of the emission is studied with respect to the dimensions of the antenna, and it is observed that the capacitance of the LAPCA leads to a severe decrease in and distortion of the biasing voltage pulse, fundamentally limiting the maximum applied bias field and consequently the maximum energy of the radiated THz pulses. In order to demonstrate the advantages of this source in the strong field regime, an open-aperture Z-scan experiment was performed on n-doped InGaAs, which showed significant absorption bleaching. PMID:27410061

  3. Study on the method to test large-aperture hyperboloid convex mirror

    NASA Astrophysics Data System (ADS)

    Meng, Xiaohui; Dong, Huiwen; Guo, Wen; Wang, Huijun

    2014-08-01

    There are numerous reflecting optical system designs that call for large-aperture convex surfaces, such as secondary mirror in on-axis three mirror anastigmatic (TMA). Several methods to test high accuracy hyperboloid convex surfaces are introduced separately in this paper. A kind of arrangement is chosen to test a surface with diameter of 420mm, radius of 1371mm, and conic K -2.1229. The CGH compensator for testing is designed, which is made up of illumination lens and hologram test plate with designed residual wavefront aberration less than 0.001λ (RMS). The second transmitted method that is equipped with a technical flat surface coating by Ag film in the bottom of surface mirror under test, which form an auto-collimation optical system to eliminate the aberration. The Hindle-Simpson test that requires a larger meniscus lens to compensate the optical aberration, and the designed result of optical test system is less than 0.0016λ. Contrasting the CGH compensator and the second transmitted method, the Hindle-Simpson testing method has the advantage of it is easily to manufacture and adjust; meanwhile the test result is stable and has been less affected by the environment. It has been found that the method is rational and reliable, and it can fulfill the requirement of manufacturing and testing process for hyperboloid convex mirrors.

  4. Development of Large-Aperture, Light-Weight Fresnel Lenses for Gossamer Space Telescopes

    SciTech Connect

    Sham, D; Hyde, R; Weisberg, A; Early, J; Rushford, M; Britten, J

    2002-04-29

    In order to examine more distant astronomical objects, with higher resolution, future space telescopes require objectives with significantly larger aperture than presently available. NASA has identified a progression in size from the 2.4m aperture objective currently used in the HUBBLE space telescope[l,2], to 25m and greater in order to observe, e.g., extra-solar planets. Since weight is a crucial factor for any object sent into space, the relative weight of large optics over a given area must be reduced[3]. The areal mass density of the primary mirror for the Hubble space telescope is {approx}200 kg/m{sup 2}. This is expected to be reduced to around 15 kg/m{sup 2} for the successor to Hubble--the next generation space telescope (NGST)[4]. For future very large aperture telescopes needed for extra-solar planet detection, the areal mass density must be reduced even further. For example, the areal mass density goal for the Gossamer space telescopes is < 1 kg/m{sup 2}. The production of lightweight focusing optics at >10m size is also an enabling technology for many other applications such as Earth observation, power beaming, and optical communications.

  5. The Advanced Technology Large-Aperture Space Telescope (ATLAST) Technology Roadmap

    NASA Technical Reports Server (NTRS)

    Stahle, Carl; Balasubramanian, K.; Bolcar, M.; Clampin, M.; Feinberg, L.; Hartman, K.; Mosier, C.; Quijada, M.; Rauscher, B.; Redding, D.; Shaklan, S.; Stahl, P.; Thronson, H.

    2014-01-01

    We present the key technologies and capabilities that will enable a future, large-aperture ultravioletopticalinfrared (UVOIR) space observatory. These include starlight suppression systems, vibration isolation and control systems, lightweight mirror segments, detector systems, and mirror coatings. These capabilities will provide major advances over current and near-future observatories for sensitivity, angular resolution, and starlight suppression. The goals adopted in our study for the starlight suppression system are 10-10 contrast with an inner working angle of 40 milliarcsec and broad bandpass. We estimate that a vibration and isolation control system that achieves a total system vibration isolation of 140 dB for a vibration-isolated mass of 5000 kg is required to achieve the high wavefront error stability needed for exoplanet coronagraphy. Technology challenges for lightweight mirror segments include diffraction-limited optical quality and high wavefront error stability as well as low cost, low mass, and rapid fabrication. Key challenges for the detector systems include visible-blind, high quantum efficiency UV arrays, photon counting visible and NIR arrays for coronagraphic spectroscopy and starlight wavefront sensing and control, and detectors with deep full wells with low persistence and radiation tolerance to enable transit imaging and spectroscopy at all wavelengths. Finally, mirror coatings with high reflectivity ( 90), high uniformity ( 1) and low polarization ( 1) that are scalable to large diameter mirror substrates will be essential for ensuring that both high throughput UV observations and high contrast observations can be performed by the same observatory.

  6. A conceptual design for a Cassegrain-mounted high-resolution optical spectrograph for large-aperture telescopes

    NASA Astrophysics Data System (ADS)

    Froning, Cynthia S.; Osterman, Steven; Burgh, Eric; Beasley, Matthew; Scowen, Paul; Veach, Todd; Jordan, Steven; Ebbets, Dennis; Lieber, Michael; deCino, James; Castilho, Bruno Vaz; Gneiding, Clemens; César de Oliveira, Antonio

    2013-09-01

    We present a conceptual design for a high-resolution optical spectrograph appropriate for mounting at Cassegrain on a large aperture telescope. The design is based on our work for the Gemini High Resolution Optical Spectrograph (CUGHOS) project. Our design places the spectrograph at Cassegrain focus to maximize throughput and blue wavelength coverage, delivering R=40,000 resolving power over a continuous 320-1050 nm waveband with throughputs twice those of current instruments. The optical design uses a two-arm, cross-dispersed echelle format with each arm optimized to maximize efficiency. A fixed image slicer is used to minimize optics sizes. The principal challenge for the instrument design is to minimize flexure and degradation of the optical image. To ensure image stability, our opto-mechanical design combines a cost-effective, passively stable bench employing a honeycomb aluminum structure with active flexure control. The active flexure compensation consists of hexapod mounts for each focal plane with full 6-axis range of motion capability to correct for focus and beam displacement. We verified instrument performance using an integrated model that couples the optical and mechanical design to image performance. The full end-to-end modeling of the system under gravitational, thermal, and vibrational perturbations shows that deflections of the optical beam at the focal plane are <29 μm per exposure under the worst case scenario (<10 μm for most orientations), with final correction to 5 μm or better using open-loop active control to meet the stability requirement. The design elements and high fidelity modeling process are generally applicable to instruments requiring high stability under a varying gravity vector.

  7. Application of image entropy evaluation function for the leveling of large aperture components in auto defects detecting

    NASA Astrophysics Data System (ADS)

    Cao, Pin; Liu, Dong; Zhao, Peng; Yang, Yong-ying; Wang, Shi-tong

    2012-10-01

    In large aperture component's dark-field scattering defects imaging system, the component's size is large and part with a wedge. When the component is in the completely level position, the surface defects image can be clearly acquired by a high magnification microscope. Otherwise, fuzzy defects image would be gained because of defocusing which makes digital identification can't be able to be done. For the problem of leveling large aperture, wedge component, this paper proposes a method that using image information entropy as focusing evaluation function for leveling large aperture components. Firstly, in three different points of component surface acquiring multi-images by the same continuous steps. Then calculating the images' entropy and fitting a curve to it. Based on minimum image information entropy value criterion, the focal plane can be found and each point's defocusingamount of the fist acquisition position can be gained. Relay on the relation model of acquisition points, adjust points and defocusingamount that has been built, each adjust point's adjustment can be got. The component's level position can be achieved by adjusting the adjust points. In the experiment that using a high magnification (of 16) microscope scans over the whole surface of the component with the size of 430mm×430mm. The image microscope is always in the depth of focus which shows that the leveling precision has achieved 20μm. Until now, this method has been successfully used in large aperture component's dark-field scattering defects imaging system.

  8. Large-aperture, tapered fiber-coupled, 10-kHz particle-image velocimetry.

    PubMed

    Hsu, Paul S; Roy, Sukesh; Jiang, Naibo; Gord, James R

    2013-02-11

    We demonstrate the design and implementation of a fiber-optic beam-delivery system using a large-aperture, tapered step-index fiber for high-speed particle-image velocimetry (PIV) in turbulent combustion flows. The tapered fiber in conjunction with a diffractive-optical-element (DOE) fiber-optic coupler significantly increases the damage threshold of the fiber, enabling fiber-optic beam delivery of sufficient nanosecond, 532-nm, laser pulse energy for high-speed PIV measurements. The fiber successfully transmits 1-kHz and 10-kHz laser pulses with energies of 5.3 mJ and 2 mJ, respectively, for more than 25 min without any indication of damage. It is experimentally demonstrated that the tapered fiber possesses the high coupling efficiency (~80%) and moderate beam quality for PIV. Additionally, the nearly uniform output-beam profile exiting the fiber is ideal for PIV applications. Comparative PIV measurements are made using a conventionally (bulk-optic) delivered light sheet, and a similar order of measurement accuracy is obtained with and without fiber coupling. Effective use of fiber-coupled, 10-kHz PIV is demonstrated for instantaneous 2D velocity-field measurements in turbulent reacting flows. Proof-of-concept measurements show significant promise for the performance of fiber-coupled, high-speed PIV using a tapered optical fiber in harsh laser-diagnostic environments such as those encountered in gas-turbine test beds and the cylinder of a combustion engine. PMID:23481818

  9. [Research of spectrum signal-to-noise ratio of large aperture static imaging spectrometer].

    PubMed

    Wang, Shuang; Li, Li-Bo; Pi, Hai-Feng

    2014-03-01

    The process of acquiring hyperspectral data cube of a Large Aperture Static Imaging Spectrometer (LASIS) includes several vital and essential steps, such as interferometer modulation, rectangular convolution sampling by pixels of detector and spectra retrieving. In this process, how to precisely evaluate the Signal-Noise Ratio (SNR) of spectra and how to wholly establish a related evaluation model were both generally very complicated. After a full consideration of the transmission process, utilizing the theory of rectangular convolution sampling and the spectral retrieving method regarding the computation of real part of the discrete Fourier transform of interferogram, formulas of both spectral signal and spectral noise were deduced theoretically, and then a evaluation model regarding the spectral SNR of LASIS was established. By using this model and other design factors of LASIS involving the wavenumber related optical transmittance, the interferometer beam splitter efficiency, the detector quantum efficiency and the main circuit noise, a simulation of spectral SNR was implemented. The simulation result was compared with the measurement result of the SNR of a LASIS instrument. The SNR lines and trends of the two match each other basically in single spectral band. The average deviation between them is proved to be 3.58%. This comparison result demonstrates the feasibility and effectiveness of the evaluation model. This SNR evaluation model consisting of the main technical aspects of typical LASIS instrument from the input spectral radiation to the output spectrum data is possible to be applied widely in practical design and implement of LASIS, as well as may provide valuable reference on SNR calculation and evaluation for other imaging spectrometers. PMID:25208427

  10. OASIS 1.0: Very Large-Aperture High-Power Lidar for Exploring Geospace

    NASA Astrophysics Data System (ADS)

    Chu, X.; Smith, J. A.; Chen, C.; Zhao, J.; Yu, Z.; Gardner, C. S.

    2015-12-01

    A new initiative, namely OASIS (the Observatory for Atmosphere Space Interaction Studies), has called for a very large-aperture high-power (VLAHP) lidar as its first step forward to acquire the unprecedented measurement capabilities for exploring the space-atmosphere interaction region (SAIR). Currently, there exists a serious observational gap of the Earth's neutral atmosphere above 100 km. Information on neutral winds and temperatures and on the plasma-neutral coupling in the SAIR, especially between 100 and 200 km, is either sparse or nonexistent. Fully exploring the SAIR requires measurements of the neutral atmosphere to complement radar observations of the plasma. Lidar measurements of neutral winds, temperatures and species can enable these explorations. Many of these topics will be addressed with the VLAHP lidar. Discoveries of thermospheric neutral Fe, Na and K layers up to nearly 200 km at McMurdo, Antarctica and other locations on Earth, have opened a new door to observing the neutral thermosphere with ground-based instruments. These neutral metal layers provide the tracers for resonance Doppler lidars to directly measure the neutral temperatures and winds in the thermosphere, thus enabling the VLAHP lidar dream! Because the thermospheric densities of these metal atoms are many times smaller than the layer peak densities near 90 km, high power-aperture product lidars, like the VLAHP lidar, are required to derive scientifically useful measurements. Furthermore, several key technical challenges for VLAHP lidar have been largely resolved in the last a few years through the successful development of Fe and Na Doppler lidars at Boulder. By combining Rayleigh and Raman with resonance lidar techniques and strategically operating the VLAHP lidar next to incoherent scatter radar and other complementary instruments, the VLAHP lidar will enable new cutting-edge exploration of the geospace. These new concepts and progresses will be introduced in this paper.

  11. Tethered Formation Configurations: Meeting the Scientific Objectives of Large Aperture and Interferometric Science

    NASA Technical Reports Server (NTRS)

    Farley, Rodger E.; Quinn, David A.; Brodeur, Stephen J. (Technical Monitor)

    2001-01-01

    With the success of the Hubble Space Telescope, it has become apparent that new frontiers of science and discovery are made every time an improvement in imaging resolution is made. For the HST working primarily in the visible and near-visible spectrum, this meant designing, building, and launching a primary mirror approximately three meters in diameter. Conventional thinking tells us that accomplishing a comparable improvement in resolution at longer wavelengths for Earth and Space Science applications requires a corresponding increase in the size of the primary mirror. For wavelengths in the sub-millimeter range, a very large telescope with an effective aperture in excess of one kilometer in diameter would be needed to obtain high quality angular resolution. Realistically a single aperture this large is practically impossible. Fortunately such large apertures can be constructed synthetically. Possibly as few as three 34 meter diameter mirrors flying in precision formation could be used to collect light at these longer wavelengths permitting not only very large virtual aperture science to be carried out, but high-resolution interferometry as well. To ensure the longest possible mission duration, a system of tethered spacecraft will be needed to mitigate the need for a great deal of propellant. A spin-stabilized, tethered formation will likely meet these requirements. Several configurations have been proposed which possibly meet the needs of the Space Science community. This paper discusses two of them, weighing the relative pros and cons of each concept. The ultimate goal being to settle on a configuration which combines the best features of structure, tethers, and formation flying to meet the ambitious requirements necessary to make future large synthetic aperture and interferometric science missions successful.

  12. Tethered Formation Configurations: Meeting the Scientific Objectives of Large Aperture and Interferometric Science

    NASA Technical Reports Server (NTRS)

    Farley, Rodger E.; Quinn, David A.

    2004-01-01

    With the success of the Hubble Space Telescope, it has become apparent that new frontiers of science and discovery are made every time an improvement in imaging resolution is made. For the HST working primarily in the visible and near-visible spectrum, this meant designing, building and launching a primary mirror approximately three meters in diameter. Conventional thinking tells us that accomplishing a comparable improvement in resolution at longer wavelengths for Earth and Space Science applications requires a corresponding increase in the size of the primary mirror. For wavelengths in the sub-millimeter range, a very large telescope with an effective aperture in excess of one kilometer in diameter would be needed to obtain high quality angular resolution. Realistically a single aperture this large is practically impossible. Fortunately such large apertures can be constructed synthetically. Possibly as few as three 3 - 4 meter diameter mirrors flying in precision formation could be used to collect light at these longer wavelengths permitting not only very large virtual aperture science to be carried out, but high-resolution interferometry as well. To ensure the longest possible mission duration, a system of tethered spacecraft will be needed to mitigate the need for a great deal of propellant. A spin-stabilized, tethered formation will likely meet these requirements. Several configurations have been proposed which possibly meet the needs of the Space Science community. This paper discusses two of them, weighing the relative pros and cons of each concept. The ultimate goal being to settle on a configuration which combines the best features of structure, tethers and formation flying to meet the ambitious requirements necessary to make future large synthetic aperture and interferometric science missions successful.

  13. HI at z 20: The Large Aperture Experiment to Detect the Dark Ages

    NASA Astrophysics Data System (ADS)

    Greenhill, Lincoln J.; Werthimer, D.; Taylor, G.; Ellingson, S.; LEDA Collaboration

    2012-05-01

    When did the first stars form? Did supermassive black holes form at the same time, earlier, or later? One of the great challenges of cosmology today is the study of these first generation objects. The Large Aperture Experiment to Detect the Dark Ages (LEDA) project seeks to detect, in total-power, emission from neutral Hydrogen (21 cm rest wavelength) in the intergalactic medium about 100 million years after the Big Bang (redshifts 20). Detection would deliver the first observational constraints on models of structure formation and the first pockets of star and black holes formation in the Universe. LEDA will develop and integrate by 2013 signal processing instrumentation into the new first station of the Long Wavelength Array (LWA). This comprises a large-N correlator serving all 512 dipole antennas of the LWA1, leveraging a packetized CASPER architecture and combining FPGAs and GPUs for the F and X stages. Iterative calibration and imaging will rely on warped snapshot imaging and be drawn from a GPU-enabled library (cuWARP) that is designed specifically to support wide-field full polarization imaging with fixed dipole arrays. Calibration techniques will include peeling, correction for ionospheric refraction, direction dependent dipole gains, deconvolution via forward modeling, and exploration of pulsar data analysis to improve performance. Accurate calibration and imaging will be crucial requirements for LEDA, necessary to subtract the bright foreground sky and detect the faint neutral Hydrogen signal. From the computational standpoint, LEDA is a O(100) TeraFlop per second challenge that enables a scalable architecture looking toward development of radio arrays requiring power efficient 10 PetaFlop per second performance. Stage two of the Hydrogen Epoch of Reionization Array (HERA2) is one example.

  14. Assessing Inter-Sensor Variability and Sensible Heat Flux Derivation Accuracy for a Large Aperture Scintillometer

    PubMed Central

    Rambikur, Evan H.; Chávez, José L.

    2014-01-01

    The accuracy in determining sensible heat flux (H) of three Kipp and Zonen large aperture scintillometers (LAS) was evaluated with reference to an eddy covariance (EC) system over relatively flat and uniform grassland near Timpas (CO, USA). Other tests have revealed inherent variability between Kipp and Zonen LAS units and bias to overestimate H. Average H fluxes were compared between LAS units and between LAS and EC. Despite good correlation, inter-LAS biases in H were found between 6% and 13% in terms of the linear regression slope. Physical misalignment was observed to result in increased scatter and bias between H solutions of a well-aligned and poorly-aligned LAS unit. Comparison of LAS and EC H showed little bias for one LAS unit, while the other two units overestimated EC H by more than 10%. A detector alignment issue may have caused the inter-LAS variability, supported by the observation in this study of differing power requirements between LAS units. It is possible that the LAS physical misalignment may have caused edge-of-beam signal noise as well as vulnerability to signal noise from wind-induced vibrations, both having an impact on the solution of H. In addition, there were some uncertainties in the solutions of H from the LAS and EC instruments, including lack of energy balance closure with the EC unit. However, the results obtained do not show clear evidence of inherent bias for the Kipp and Zonen LAS to overestimate H as found in other studies. PMID:24473285

  15. Enzyme Activities in Polarized Cell Membranes

    PubMed Central

    Bass, L.; McIlroy, D. K.

    1968-01-01

    The theoretical pH dependence of enzyme activities in membranes of low dielectric constant is estimated. It is shown that in biological membranes some types of enzymes may attain a limiting pH sensitivity such that an increment of only 0.2 pH unit (sufficient to induce action potentials in squid axons) causes a relative activity change of over 25%. The transients of enzyme activity generated by membrane depolarization and by pH increments in the bathing solution are discussed in relation to the transients of nervous excitation. PMID:5641405

  16. Versatile Membrane Deformation Potential of Activated Pacsin

    PubMed Central

    Byrnes, Laura J.; Sondermann, Holger

    2012-01-01

    Endocytosis is a fundamental process in signaling and membrane trafficking. The formation of vesicles at the plasma membrane is mediated by the G protein dynamin that catalyzes the final fission step, the actin cytoskeleton, and proteins that sense or induce membrane curvature. One such protein, the F-BAR domain-containing protein pacsin, contributes to this process and has been shown to induce a spectrum of membrane morphologies, including tubules and tube constrictions in vitro. Full-length pacsin isoform 1 (pacsin-1) has reduced activity compared to its isolated F-BAR domain, implicating an inhibitory role for its C-terminal Src homology 3 (SH3) domain. Here we show that the autoinhibitory, intramolecular interactions in pacsin-1 can be released upon binding to the entire proline-rich domain (PRD) of dynamin-1, resulting in potent membrane deformation activity that is distinct from the isolated F-BAR domain. Most strikingly, we observe the generation of small, homogenous vesicles with the activated protein complex under certain experimental conditions. In addition, liposomes prepared with different methods yield distinct membrane deformation morphologies of BAR domain proteins and apparent activation barriers to pacsin-1's activity. Theoretical free energy calculations suggest bimodality of the protein-membrane system as a possible source for the different outcomes, which could account for the coexistence of energetically equivalent membrane structures induced by BAR domain-containing proteins in vitro. Taken together, our results suggest a versatile role for pacsin-1 in sculpting cellular membranes that is likely dependent both on protein structure and membrane properties. PMID:23236520

  17. Imaging with Concave Large-Aperture Therapeutic Ultrasound Arrays Using Conventional Synthetic-Aperture Beamforming

    PubMed Central

    Wan, Yayun; Ebbini, Emad S.

    2009-01-01

    Several dual-mode ultrasound array (DMUA) systems are being investigated for potential use in image-guided surgery. In therapeutic mode, DMUAs generate pulsed or continuous-wave (CW) high-intensity focused ultrasound (HIFU) beams capable of generating localized therapeutic effects within the focal volume. In imaging mode, pulse-echo data can be collected from the DMUA elements to obtain B-mode images or other forms of feedback on the state of the target tissue before, during, and after the application of the therapeutic HIFU beam. Therapeutic and technological constraints give rise to special characteristics of therapeutic arrays. Specifically, DMUAs have concave apertures with low f-number values and are typically coarsely sampled using directive elements. These characteristics necessitate pre- and post-beamforming signal processing of echo data to improve the spatial and contrast resolution and maximize the image uniformity within the imaging field of view (IxFOV). We have recently developed and experimentally validated beamforming algorithms for concave large-aperture DMUAs with directive elements. Experimental validation was performed using a 1 MHz, 64-element, concave spherical aperture with 100 mm radius of curvature. The aperture was sampled in the lateral direction using elongated elements 1−λ×33.3‒ with 1.333‒−λ center-to-center spacing (λ is the wavelength). This resulted in f-number values of 0.8 and 2 in the azimuth and elevation directions, respectively. In this paper, we present a new DMUA design approach based on different sampling of the shared concave aperture to improve image quality while maintaining therapeutic performance. A pulse-wave (PW) simulation model using a modified version of the Field II program is used in this study. The model is used in generating pulse-echo data for synthetic-aperture (SA) beamforming for forming images of a variety of targets, e.g., wire arrays and speckle-generating cyst phantoms. To provide

  18. Research on 2x1 plasma electrode electro-optical switch with large aperture

    NASA Astrophysics Data System (ADS)

    Zhang, Xiong Jun; Zheng, Kui Xing; Feng, B.; Wu, D. S.; Lu, J. P.; Tian, X. L.; Jin, F.; Sui, Zhan; Wei, Xiaofeng; Zhang, Xiaomin

    2005-01-01

    In conceptual design of the prototype for SG-III facility, a full aperture electro-optical switch was placed between the cavity mirror and the main amplifier to isolate the reflected beams. The beam on the cavity mirror is 240mm×240mm square. Pockells cells of conversional design with coaxial ring electrodes can not scale to such large square aperture. In the 1980s, a plasma electrode Pockels cell (PEPC) concept was invented at LLNL. It uses transparent plasma electrode formed through gas discharge as the electrodes to apply the voltage across switching crystal to rotate the polarization of a transmitted laser beam. And it can be scaled to large aperture with thin crystal. So the switch which would be used in SG-III is based on this technology. The technical integration line as a prototype of SG-III laser is actually a 4×2 beam bundle. And the full aperture optical switch is mechanically designed four apertures as a removable unit, and electrically two 2×1 PEPC putting together. So we built a 2×1 PEPC to develop the technology first. The 2×1 PEPC is a sandwich structure made of an insulating mid plane between a pair of plasma chambers. The frame of both plasma chambers are machining in duralumin. Each chamber is installed with a planar magnetic cathode and four segments spherical anodes made from stainless steel. The cathode and anode are insulated from the housing with a special shell made from plastic, and plasma is insulated from the housing by an 80-μm-thick anodic coating on the duralumin. The two plasma chambers are separated by a mid plane of glass frame with two square holes. The two holes are filled by two electro-optical crystals with a 240-mm square aperture. With the optimized operating pressure and the electrical parameters, a very good homogeneity and low resistivity plasma electrode is obtained. Finally we tested its switching performance to simulate the case that it will be used in the SG-III prototype facility. It works with a quarter wave

  19. Determining suitability of Large Aperture Scintillometer for validating remote sensing based evapotranspiration maps

    NASA Astrophysics Data System (ADS)

    Paul, G.; Gowda, P. H.; Howell, T. A.; Basu, S.; Colaizzi, P. D.; Marek, T.

    2013-12-01

    Scintillation method is a relatively new technique for measuring the sensible heat and water fluxes over land surfaces. Path integrating capabilities of scintillometer over heterogeneous landscapes make it a potential tool for comparing the energy fluxes derived from remote sensing based energy balance algorithms. For this reason, scintillometer-derived evapotranspiration (ET) fluxes are being used to evaluate remote sensing based energy balance algorithms for their ability to estimate ET fluxes. However, LAS' (Large Aperture Scintillometer) ability to derive ET fluxes is not thoroughly tested. The objective of this study was to evaluate LAS- and Surface Energy Balance System (SEBS)-derived fluxes against lysimetric data to determine LAS' suitability for validating remote sensing based evapotranspiration (ET) maps. The study was conducted during the Bushland Evapotranspiration and Agricultural Remote sensing EXperiment - 2008 (BEAREX-08) at the USDA-ARS Conservation and Production Research Laboratory (CPRL), Bushland, Texas. SEBS was coded in a GIS environment to retrieve ET fluxes from the high resolution imageries acquired using airborne multispectral sensors. The CPRL has four large weighing lysimeters (3 m long x 3 m wide x 2.4 m deep), each located in the middle of approximately 5 ha fields, arranged in a block pattern. The two lysimeter fields located on the east (NE and SE) were managed under irrigated conditions, and the other two lysimeters on the west (NW and SW) were under dryland management. Each lysimeter field was equipped with an automated weather station that provided measurements for net radiation (Rn), Ts, soil heat flux (Go), Ta, relative humidity, and wind speed. During BEAREX08, the NE and SE fields were planted to cotton on May 21, and the NW and SW dryland lysimeters fields were planted to cotton on June 5. One LAS each was deployed across two large dryland lysimeter fields (NW and SW) and two large irrigated lysimeter fields (NE and SE). The

  20. [Design of a dual-channel Mach-Zehnder lateral shearing interferometer for the large aperture static imaging spectrometer].

    PubMed

    Fu, Qiang; Xiangli, Bin; Lü, Qun-bo; Jing, Juan-juan

    2012-02-01

    Large aperture static imaging spectrometry (LASIS) is a kind of joint temporally and spatially modulated Fourier transform imaging spectrometry. In such instruments, lateral shearing interferometer is a key element, the most frequently used type of which is the Sagnac interferometer. In this configuration, one half of the light entering the interferometer backtracks and causes a great decrease in energy efficiency. The present paper proposes a modified Mach-Zehnder lateral shearing interferometer structure to tackle this problem. With the ability to produce the same lateral shear, it features the advantage of dual channel output. We present a ray tracing procedure to induce the general expression of the lateral shear as well as analyze the contributions of error sources to the shear accuracy. The results serve as a new idea for the design of large aperture static imaging spectrometers and can be used to instruct the design and optimization of this kind of imaging spectrometer. PMID:22512210

  1. A novel measurement scheme for the radial group delay of large-aperture ultra-short laser pulses

    NASA Astrophysics Data System (ADS)

    Wu, Fenxiang; Xu, Yi; Li, Zhaoyang; Li, Wenkai; Lu, Jun; Wang, Cheng; Li, Yanyan; Liu, Yanqi; Lu, Xiaoming; Peng, Yujie; Wang, Ding; Leng, Yuxin; Li, Ruxin

    2016-05-01

    In femtosecond high-peak-power laser system, the radial group delay (RGD) of the pulse front introduced by conventional lens-based beam expanders can significantly decrease the achievable focal intensity, especially when it is larger than the pulse duration. In order to quantitatively analyze and compensate the RGD, a novel measurement scheme based on self-reference and second-order cross-correlation technology is proposed and applied to measure the RGD of the large-aperture ultra-short laser pulses directly. The measured result of the RGD in a 200 TW Ti:sapphire laser system is in good agreement with the theoretical calculation. To our knowledge, it is the first time to realize the direct RGD measurement of large-aperture ultra-short laser pulses.

  2. Active Nuclear Import of Membrane Proteins Revisited

    PubMed Central

    Laba, Justyna K.; Steen, Anton; Popken, Petra; Chernova, Alina; Poolman, Bert; Veenhoff, Liesbeth M.

    2015-01-01

    It is poorly understood how membrane proteins destined for the inner nuclear membrane pass the crowded environment of the Nuclear Pore Complex (NPC). For the Saccharomyces cerevisiae proteins Src1/Heh1 and Heh2, a transport mechanism was proposed where the transmembrane domains diffuse through the membrane while the extralumenal domains encoding a nuclear localization signal (NLS) and intrinsically disordered linker (L) are accompanied by transport factors and travel through the NPC. Here, we validate the proposed mechanism and explore and discuss alternative interpretations of the data. First, to disprove an interpretation where the membrane proteins become membrane embedded only after nuclear import, we present biochemical and localization data to support that the previously used, as well as newly designed reporter proteins are membrane-embedded irrespective of the presence of the sorting signals, the specific transmembrane domain (multipass or tail anchored), independent of GET, and also under conditions that the proteins are trapped in the NPC. Second, using the recently established size limit for passive diffusion of membrane proteins in yeast, and using an improved assay, we confirm active import of polytopic membrane protein with extralumenal soluble domains larger than those that can pass by diffusion on similar timescales. This reinforces that NLS-L dependent active transport is distinct from passive diffusion. Thirdly, we revisit the proposed route through the center of the NPC and conclude that the previously used trapping assay is, unfortunately, poorly suited to address the route through the NPC, and the route thus remains unresolved. Apart from the uncertainty about the route through the NPC, the data confirm active, transport factor dependent, nuclear transport of membrane-embedded mono- and polytopic membrane proteins in baker’s yeast. PMID:26473931

  3. Active Nuclear Import of Membrane Proteins Revisited.

    PubMed

    Laba, Justyna K; Steen, Anton; Popken, Petra; Chernova, Alina; Poolman, Bert; Veenhoff, Liesbeth M

    2015-01-01

    It is poorly understood how membrane proteins destined for the inner nuclear membrane pass the crowded environment of the Nuclear Pore Complex (NPC). For the Saccharomyces cerevisiae proteins Src1/Heh1 and Heh2, a transport mechanism was proposed where the transmembrane domains diffuse through the membrane while the extralumenal domains encoding a nuclear localization signal (NLS) and intrinsically disordered linker (L) are accompanied by transport factors and travel through the NPC. Here, we validate the proposed mechanism and explore and discuss alternative interpretations of the data. First, to disprove an interpretation where the membrane proteins become membrane embedded only after nuclear import, we present biochemical and localization data to support that the previously used, as well as newly designed reporter proteins are membrane-embedded irrespective of the presence of the sorting signals, the specific transmembrane domain (multipass or tail anchored), independent of GET, and also under conditions that the proteins are trapped in the NPC. Second, using the recently established size limit for passive diffusion of membrane proteins in yeast, and using an improved assay, we confirm active import of polytopic membrane protein with extralumenal soluble domains larger than those that can pass by diffusion on similar timescales. This reinforces that NLS-L dependent active transport is distinct from passive diffusion. Thirdly, we revisit the proposed route through the center of the NPC and conclude that the previously used trapping assay is, unfortunately, poorly suited to address the route through the NPC, and the route thus remains unresolved. Apart from the uncertainty about the route through the NPC, the data confirm active, transport factor dependent, nuclear transport of membrane-embedded mono- and polytopic membrane proteins in baker's yeast. PMID:26473931

  4. Thioredoxin-like activity of thylakoid membranes

    SciTech Connect

    Ashton, A.R.; Brennan, T.; Anderson, L.E.

    1980-10-01

    The inactivation of pea leaf chloroplast glucose-6-phosphate dehydrogenase by dithiothreitol can be catalyzed by thioredoxin-like molecules that are present in chloroplasts. This thioredoxin activity occurs predominantly as a soluble species, but washed thylakoid membranes also exhibit some thioredoxin-like activity. The membrane-associated thioredoxin can be extracted by treatment with the detergent Triton X-100. The solubilized thioredoxing appears to have a molecular size similar to that of the soluble thioredoxin which catalyzes the same reaction. The thylakoid-bound activity constitutes only about 5% of the total chloroplast thioredoxin activity. The thioredoxin occurring in the membrane fraction cannot, however, be ascribed to the trapping of stroma since less than 0.1% of three stromal marker enzymes are found in the same thylakoid extract.

  5. Organelle morphogenesis by active membrane remodeling

    NASA Astrophysics Data System (ADS)

    Ramakrishnan, N.; Ipsen, John H.; Rao, Madan; Kumar, P. B. Sunil

    Intracellular organelles are subject to a steady flux of lipids and proteins through active, energy consuming transport processes. Active fission and fusion are promoted by GTPases, e.g., Arf-Coatamer and the Rab-Snare complexes, which both sense and generate local membrane curvature. Here we investigate through Dynamical Triangulation Monte Carlo simulations, the role that these active processes play in determining the morphology and compositional segregation in closed membranes. Our results suggest that the ramified morphologies of organelles observed in-vivo are a consequence of driven nonequilibrium processes rather than equilibrium forces.

  6. Active membrane cholesterol as a physiological effector.

    PubMed

    Lange, Yvonne; Steck, Theodore L

    2016-09-01

    Sterols associate preferentially with plasma membrane sphingolipids and saturated phospholipids to form stoichiometric complexes. Cholesterol in molar excess of the capacity of these polar bilayer lipids has a high accessibility and fugacity; we call this fraction active cholesterol. This review first considers how active cholesterol serves as an upstream regulator of cellular sterol homeostasis. The mechanism appears to utilize the redistribution of active cholesterol down its diffusional gradient to the endoplasmic reticulum and mitochondria, where it binds multiple effectors and directs their feedback activity. We have also reviewed a broad literature in search of a role for active cholesterol (as opposed to bulk cholesterol or lipid domains such as rafts) in the activity of diverse membrane proteins. Several systems provide such evidence, implicating, in particular, caveolin-1, various kinds of ABC-type cholesterol transporters, solute transporters, receptors and ion channels. We suggest that this larger role for active cholesterol warrants close attention and can be tested easily. PMID:26874289

  7. Reconstitution of Membrane Proteins into Model Membranes: Seeking Better Ways to Retain Protein Activities

    PubMed Central

    Shen, Hsin-Hui; Lithgow, Trevor; Martin, Lisandra L.

    2013-01-01

    The function of any given biological membrane is determined largely by the specific set of integral membrane proteins embedded in it, and the peripheral membrane proteins attached to the membrane surface. The activity of these proteins, in turn, can be modulated by the phospholipid composition of the membrane. The reconstitution of membrane proteins into a model membrane allows investigation of individual features and activities of a given cell membrane component. However, the activity of membrane proteins is often difficult to sustain following reconstitution, since the composition of the model phospholipid bilayer differs from that of the native cell membrane. This review will discuss the reconstitution of membrane protein activities in four different types of model membrane—monolayers, supported lipid bilayers, liposomes and nanodiscs, comparing their advantages in membrane protein reconstitution. Variation in the surrounding model environments for these four different types of membrane layer can affect the three-dimensional structure of reconstituted proteins and may possibly lead to loss of the proteins activity. We also discuss examples where the same membrane proteins have been successfully reconstituted into two or more model membrane systems with comparison of the observed activity in each system. Understanding of the behavioral changes for proteins in model membrane systems after membrane reconstitution is often a prerequisite to protein research. It is essential to find better solutions for retaining membrane protein activities for measurement and characterization in vitro. PMID:23344058

  8. Developing Magnetorheological Finishing (MRF) Technology for the Manufacture of Large-Aperture Optics in Megajoule Class Laser Systems

    SciTech Connect

    Menapace, J A

    2010-10-27

    Over the last eight years we have been developing advanced MRF tools and techniques to manufacture meter-scale optics for use in Megajoule class laser systems. These systems call for optics having unique characteristics that can complicate their fabrication using conventional polishing methods. First, exposure to the high-power nanosecond and sub-nanosecond pulsed laser environment in the infrared (>27 J/cm{sup 2} at 1053 nm), visible (>18 J/cm{sup 2} at 527 nm), and ultraviolet (>10 J/cm{sup 2} at 351 nm) demands ultra-precise control of optical figure and finish to avoid intensity modulation and scatter that can result in damage to the optics chain or system hardware. Second, the optics must be super-polished and virtually free of surface and subsurface flaws that can limit optic lifetime through laser-induced damage initiation and growth at the flaw sites, particularly at 351 nm. Lastly, ultra-precise optics for beam conditioning are required to control laser beam quality. These optics contain customized surface topographical structures that cannot be made using traditional fabrication processes. In this review, we will present the development and implementation of large-aperture MRF tools and techniques specifically designed to meet the demanding optical performance challenges required in large-aperture high-power laser systems. In particular, we will discuss the advances made by using MRF technology to expose and remove surface and subsurface flaws in optics during final polishing to yield optics with improve laser damage resistance, the novel application of MRF deterministic polishing to imprint complex topographical information and wavefront correction patterns onto optical surfaces, and our efforts to advance the technology to manufacture large-aperture damage resistant optics.

  9. In-situ monitoring of surface post-processing in large aperture fused silica optics with Optical Coherence Tomography

    SciTech Connect

    Guss, G M; Bass, I l; Hackel, R P; Mailhiot, C; Demos, S G

    2008-02-08

    Optical Coherence Tomography is explored as a method to image laser-damage sites located on the surface of large aperture fused silica optics during post-processing via CO{sub 2} laser ablation. The signal analysis for image acquisition was adapted to meet the sensitivity requirements for this application. A long-working distance geometry was employed to allow imaging through the opposite surface of the 5-cm thick optic. The experimental results demonstrate the potential of OCT for remote monitoring of transparent material processing applications.

  10. Amplification of femtosecond pulses to above 1 J with large aperture Cr:LiSrAIF{sub 6} amplifiers

    SciTech Connect

    Ditmire, T.; Perry, M.D.

    1995-01-01

    The authors have developed a chirped pulse amplification system capable of producing femtosecond pulses with energy above one joule. This is accomplished by using a large aperture, flashlamp pumped Cr-LiSrAlF{sub 6} (Cr:LiSAF) amplifier. Optimum design of the 19 mm diameter amplifier results in a single pass gain of 5 with good beam quality. This amplifier produces 1.05 J pulses after compression with a width of < 125 fs at a repetition rate of 0.05 Hz.

  11. Characterisation of a large aperture steep concave parabolic mirror using SASI based on auto-collimation theory

    NASA Astrophysics Data System (ADS)

    Liu, Bingcai; Li, Bing; Tian, Ailing; Yang, Haoyu; Gao, Fen; Chen, Lei

    2015-01-01

    To characterise a large aperture steep concave parabolic mirror, a new sub-aperture stitching interferometry measurement technology (SASI) based on auto-collimation is proposed. The principle of the stitching process is analysed, and the sub-aperture partitioning for a full aperture of a paraboloid is discussed. Next, the overlapped sampled points between sub-apertures are rectified through sampled points realigned in mesh grids. Finally, two experiments, the SASI based on auto-collimation and the full aperture test, were implemented for a parabolic mirror. The stitching result exhibits good agreement with the full-aperture result.

  12. A New Type of X-ray Condenser Lenses with Large Apertures Fabricated by Rolling of Structured Films

    SciTech Connect

    Simon, M.; Reznikova, E.; Nazmov, V.; Grund, T.; Last, A.

    2010-04-06

    In order to meet the demand for X-ray lenses with large apertures and, hence, photon flux, a new type of X-ray lenses has been developed: Rolled prismatic X-ray lenses feature a vast number of refracting surfaces to increase transparency and aperture, respectively. Prototypes of such lenses have been fabricated by molding and rolling of a structured polyimide film. In this work, rolled prismatic X-ray lenses are pictured, and results of first tests performed at the ANKA storage ring in Karlsruhe are presented.

  13. Membrane optical activity: some facts and fallacies.

    PubMed

    Wallach, D F; Low, D A; Bertland, A V

    1973-11-01

    The circular dichroism of hypothetical, water-filled, spherical shells, 75-3500 nm in radius, with walls 7.5 nm thick, composed of poly(L-lysine) in various conformational proportions, and suspended in water, were computed from the known optical properties of this polypeptide by classical general light-scattering theory (Mie theory). Comparison of the computed curves of circular dichroism spectra with those of diverse membranes reveals large discrepancies below 215 nm and shows that light scattering does not adequately account for the optical activity of membranes containing appreciable proportions of nonhelical conformation. However, turbidity effects can explain the anomalies of membrane optical rotatory dispersion near 233 nm, if not uniquely so. We conclude that the optical activity of neither most soluble proteins nor membrane proteins can provide accurate conformational information when synthetic polypeptides are used as standards and list the reasons for this argument. We also show that present techniques to "correct" membrane optical activity are likely to produce additional artifact. PMID:4522300

  14. A high-resolution detecting system based on machine vision for defects on large aperture and super-smooth surface

    NASA Astrophysics Data System (ADS)

    Yang, Yongying; Zhao, Limin; Wang, Shitong; Cao, Pin; Liu, Dong; Li, Lu; Yan, Lu; Li, Chen; Xie, Shibing; Li, Yang; Chen, Yangjie

    2015-02-01

    The high-resolution detecting system based on machine vision for defects on large aperture and super-smooth surface uses a novel ring telecentric lighting optical system detecting the defects on the sample all round and without blind spots. The scattering light induced by surface defects enters the adaptive and highly zoom microscopic scattering dark-field imaging system for defect detecting and then forms digital images. Sub-aperture microscopic scanning sampling and fast stitching on the surface is realized by using precise multi-axis shifting guided scanning system and a standard comparison board based upon binary optics is used to implement fast calibration of micron-dimension defects detected actually. The pattern recognition technology of digital image processing which can automatically output digitalized surface defects statements after scaling is established to comprehensively evaluate defects. This system which can reach micron-dimension defect resolution can achieve detections of large aperture components of 850 mm × 500 mm, solve the durable problem of subjective uncertainty brought in by human visual detection of defects and achieve quantitative detection of defects with machine vision.

  15. Coherent polarization stabilization in large-aperture rectangular post bottom-emitting vertical-cavity surface-emitting lasers

    NASA Astrophysics Data System (ADS)

    Wang, W.; Ning, Y. Q.; Tian, Z. H.; Zhang, X.; Shi, J. J.; Wang, Z. F.; Zhang, L. S.; Zhang, Y.; Liu, Y.; Qin, L.; Wang, L. J.

    2011-03-01

    The output characteristics of large-aperture rectangular post bottom-emitting vertical-cavity surface-emitting lasers (VCSELs) were investigated. It was shown that the output power of the rectangle VCSELs can be up to 660 mW at a current of 5 A. Both H-polarization (horizontal) and V-polarization (vertical) demonstrated a coherent stabilization over the entire range of operation current, and coherent spectrum blue-shift of H-polarization light occurred with respect to V-polarization light at three different injected currents. The polarization states of output light were stabilized in the two orthogonal directions and H-polarization was the most principal polarization which was parallel to the longer side of the rectangular aperture. From the relationship between polarization ratio and aspect ratio of the oxidation confinement aperture (OCA), it was found that the highest polarization ratio (about 2:1) took place when the appropriate aspect ratio was 5:3, which meant better polarization stabilization in large-aperture VCSELs.

  16. Electro-Mechanical Simulation of a Large Aperture MOEMS Fabry-Perot Tunable Filter

    NASA Technical Reports Server (NTRS)

    Kuhn, Jonathan L.; Barclay, Richard B.; Greenhouse, Matthew A.; Mott, D. Brent; Satyapal, Shobita; Powers, Edward I. (Technical Monitor)

    2000-01-01

    We are developing a micro-machined electrostatically actuated Fabry-Perot tunable filter with a large clear aperture for application in high through-put wide-field imaging spectroscopy and lidar systems. In the first phase of this effort, we are developing key components based on coupled electro-mechanical simulations. In particular, the movable etalon plate design leverages high coating stresses to yield a flat surface in drum-head tension over a large diameter (12.5 mm). In this approach, the cylindrical silicon movable plate is back etched, resulting in an optically coated membrane that is suspended from a thick silicon support ring. Understanding the interaction between the support ring, suspended membrane, and coating is critical to developing surfaces that are flat to within stringent etalon requirements. In this work, we present the simulations used to develop the movable plate, spring suspension system, and electrostatic actuation mechanism. We also present results from tests of fabricated proof of concept components.

  17. Activation of Membrane Cholesterol by 63 Amphipaths†

    PubMed Central

    Lange, Yvonne; Ye, Jin; Duban, Mark-Eugene; Steck, Theodore L.

    2009-01-01

    A few membrane-intercalating amphipaths have been observed to stimulate the interaction of cholesterol with cholesterol oxidase, saponin and cyclodextrin, presumably by displacing cholesterol laterally from its phospholipid complexes. We now report that this effect, referred to as cholesterol activation, occurs with dozens of other amphipaths, including alkanols, saturated and cis- and trans-unsaturated fatty acids, fatty acid methyl esters, sphingosine derivatives, terpenes, alkyl ethers, ketones, aromatics and cyclic alkyl derivatives. The apparent potency of the agents tested ranged from 3 μM to 7 mM and generally paralleled their octanol/water partition coefficients, except that relative potency declined for compounds with> 10 carbons. Some small amphipaths activated cholesterol at a membrane concentration of ~3 moles per 100 moles bilayer lipids, about equimolar with the cholesterol they displaced. Lysophosphatidylserine countered the effects of all these agents, consistent with its ability to reduce the pool of active membrane cholesterol. Various amphipaths stabilized red cells against the hemolysis elicited by cholesterol depletion, presumably by substituting for the extracted sterol. The number and location of cis and trans fatty acid unsaturations and the absolute stereochemistry of enantiomer pairs had only small effects on amphipath potency. Nevertheless, potency varied ~7-fold within a group of diverse agents with similar partition coefficients. We infer that a wide variety of amphipaths can displace membrane cholesterol by competing stoichiometrically but with only limited specificity for its weak association with phospholipids. Any number of other drugs and experimental agents might do the same. PMID:19655814

  18. Membrane activity of biomimetic facially amphiphilic antibiotics.

    PubMed

    Arnt, Lachelle; Rennie, Jason R; Linser, Sebastian; Willumeit, Regine; Tew, Gregory N

    2006-03-01

    Membranes are a central feature of all biological systems, and their ability to control many cellular processes is critically important. As a result, a better understanding of how molecules bind to and select between biological membranes is an active area of research. Antimicrobial host defense peptides are known to be membrane-active and, in many cases, exhibit discrimination between prokaryotic and eukaryotic cells. The design of synthetic molecules that capture the biological activity of these natural peptides has been shown. In this report, the interaction between our biomimetic structures and different biological membranes is reported using both model vesicle and in vitro bacterial cell experiments. Compound 1 induces 12% leakage at 20 microg/mL against phosphatidylglycerol (PG)-phosphatidylethanolamine (PE) vesicles vs only 3% leakage at 200 microg/mL against phosphatidyl-L-serine (PS)-phosphatidylcholine (PC) vesicles. Similarly, a 40% reduction in fluorescence is measured in lipid movement experiments for PG-PE compared to 10% for PS-PC at 600 s. A 30 degrees C increase in the phase transition of stearoyl-oleoyl-phosphatidylserine is observed in the presence of 1. These results show that lipid composition is more important for selectivity than overall net charge. Additionally, the overall concentration of a given lipid is another important factor. An effort is made to connect model vesicle studies with in vitro data and naturally occurring lipid compositions. PMID:16494408

  19. Large-aperture MOEMS Fabry-Perot interferometer for miniaturized spectral imagers

    NASA Astrophysics Data System (ADS)

    Rissanen, Anna; Langner, Andreas; Viherkanto, Kai; Mannila, Rami

    2015-02-01

    VTT's optical MEMS Fabry-Perot interferometers (FPIs) are tunable optical filters, which enable miniaturization of spectral imagers into small, mass producible hand-held sensors with versatile optical measurement capabilities. FPI technology has also created a basis for various hyperspectral imaging instruments, ranging from nanosatellites, environmental sensing and precision agriculture with UAVs to instruments for skin cancer detection. Until now, these application demonstrations have been mostly realized with piezo-actuated FPIs fabricated by non-monolithical assembly method, suitable for achieving very large optical apertures and with capacity to small-to-medium volumes; however large-volume production of MEMS manufacturing supports the potential for emerging spectral imaging applications also in large-volume applications, such as in consumer/mobile products. Previously reported optical apertures of MEMS FPIs in the visible range have been up to 2 mm in size; this paper presents the design, successful fabrication and characterization of MEMS FPIs for central wavelengths of λ = 500 nm and λ = 650 nm with optical apertures up to 4 mm in diameter. The mirror membranes of the FPI structures consist of ALD (atomic layer deposited) TiO2-Al2O3 λ/4- thin film Bragg reflectors, with the air gap formed by sacrificial polymer etching in O2 plasma. The entire fabrication process is conducted below 150 °C, which makes it possible to monolithically integrate the filter structures on other ICdevices such as detectors. The realized MEMS devices are aimed for nanosatellite space application as breadboard hyperspectral imager demonstrators.

  20. The Balloon-borne Large Aperture Submillimeter Telescope for Polarimetry (BLAST-Pol): Instrument and 2010 Science Campaign

    NASA Astrophysics Data System (ADS)

    Gandilo, Natalie; BLAST-Pol Collaboration

    2012-01-01

    The Balloon-borne Large Aperture Submillimeter Telescope for Polarimetry (BLAST-Pol) is a 1.8-m telescope that observes polarized dust emission with a resolution of 1'. BLAST-Pol images the sky onto a focal plane that consists of 270 feed-horn coupled bolometers at 250, 350, and 500 microns. In January 2011, BLAST-Pol completed a successful 9.5-day flight over Antarctica. Eight science targets were observed, and a second flight is planned for December 2012. I will give an overview of the instrument performance during the first science campaign and present preliminary maps. BLAST-Pol maps will provide an excellent dataset for studying the role of magnetic fields in star formation.

  1. The Advanced Technology Large Aperture Space Telescope (ATLAST): Science Drivers, Technology Developments, and Synergies with Other Future Facilities

    NASA Technical Reports Server (NTRS)

    Postman, Marc; Brown, Tom; Sembach, Kenneth; Giavalisco, Mauro; Traub, Wesley; Stapelfeldt, Karl; Calzetti, Daniela; Oegerle, William; Rich, R. Michael; Stahl, H. Philip; Tumlinson, Jason; Mountain, Matt; Soummer, Remi; Hyde, Tupper

    2011-01-01

    The Advanced Technology Large-Aperture Space Telescope (ATLAST) is a concept for an 8-meter to 16-meter UVOIR space observatory for launch in the 2025-2030 era. ATLAST will allow astronomers to answer fundamental questions at the forefront of modern astrophysics, including "Is there life elsewhere in the Galaxy?" We present a range of science drivers that define the main performance requirements for ATLAST (8 to 16 milliarcsec angular resolution, diffraction limited imaging at 0.5 m wavelength, minimum collecting area of 45 square meters, high sensitivity to light wavelengths from 0.1 m to 2.4 m, high stability in wavefront sensing and control). We will also discuss the synergy between ATLAST and other anticipated future facilities (e.g., TMT, EELT, ALMA) and the priorities for technology development that will enable the construction for a cost that is comparable to current generation observatory-class space missions.

  2. Comparison of Turbulent Sensible Heat Flux Determined by Large-Aperture Scintillometer and Eddy Covariance over Urban and Suburban Areas

    NASA Astrophysics Data System (ADS)

    Zhang, He; Zhang, Hongsheng

    2015-01-01

    Field observations of the atmospheric boundary layer were made over urban and suburban areas in the Yangtze River Delta, China. Sensible heat fluxes were obtained by eddy-covariance (EC) systems and large-aperture scintillometers (LASs). The results indicated that (1) the sensible heat flux obtained by LAS was less noisy and slightly larger than that obtained by EC over both urban and suburban surfaces; (2) the values of were higher when the correlation coefficient of vertical wind speed and temperature () was smaller. Lower values of were due to low-frequency trends. The urban values of were smaller than suburban values at low values; (3) the sensible heat flux determined by LAS was improved by use of the Monin-Obukhov similarity theory of the temperature structure parameter over urban and suburban areas, and the improvement is more significant over urban surface areas.

  3. Design studies of large aperture, high-resolution Earth science microwave radiometers compatible with small launch vehicles

    NASA Technical Reports Server (NTRS)

    Schroeder, Lyle C.; Bailey, M. C.; Harrington, Richard F.; Kendall, Bruce M.; Campbell, Thomas G.

    1994-01-01

    High-spatial-resolution microwave radiometer sensing from space with reasonable swath widths and revisit times favors large aperture systems. However, with traditional precision antenna design, the size and weight requirements for such systems are in conflict with the need to emphasize small launch vehicles. This paper describes tradeoffs between the science requirements, basic operational parameters, and expected sensor performance for selected satellite radiometer concepts utilizing novel lightweight compactly packaged real apertures. Antenna, feed, and radiometer subsystem design and calibration are presented. Preliminary results show that novel lightweight real aperture coupled with state-of-the-art radiometer designs are compatible with small launch systems, and hold promise for high-resolution earth science measurements of sea ice, precipitation, soil moisture, sea surface temperature, and ocean wind speeds.

  4. Potential of a Future Large Aperture UVOIR Space Observatory for Breakthrough Observations of Star and Planet Formation

    NASA Astrophysics Data System (ADS)

    Danchi, William C.; Grady, Carol A.; Padgett, Deborah

    2015-01-01

    A future large aperture space observatory operating from the UV to the near-infrared with a diameter between 10 and 15 meters will provide a unique opportunity for observations of star and planet formation, from nearby moving groups and associations to star formation in galaxies in the local universe. Our newly formed working group will examine the unique opportunities that such a telescope will give observers in a post-JWST/WFIRST-AFTA era that includes extremely large ground-based observatories such as the TMT, E-ELT, ALMA, and the VLTI. Given a potential suite of instruments for this observatory we will discuss some of the key areas of star and planet formation science where breakthroughs might occur.

  5. New technologies for the actuation and controls of large aperture lightweight quality mirrors

    NASA Technical Reports Server (NTRS)

    Lih, S. S.; Yang, E. H.; Gullapalli, S. N.; Flood, R.

    2003-01-01

    This paper presents a set of candidate components: MEMS based large stroke (>100 microns) ultra lightweight (0.01 gm) discrete inch worm actuator technology, and a distributed actuator technology, in the context of a novel lightweight active flexure-hinged substrate concept that uses the nanolaminate face sheet.

  6. Distributed Bragg reflector ring oscillators: A large aperture source of high single-mode optical power

    SciTech Connect

    Dzurko, K.M.; Hardy, A.; Scifres, D.R.; Welch, D.F.; Waarts, R.G.; Lang, R.J. )

    1993-06-01

    Distributed Bragg reflector (DBR) ring oscillators are the first monolithic semiconductor lasers containing broad-area active regions which operate in a single mode to several times their threshold current. Orthogonally oriented diffraction gratings surrounding an unpatterned active region select a single spatial and temporal mode of oscillation. This paper presents both analytic and experimental verification of single mode operation for active dimensions up to 368 [times] 1000 [mu]m. Threshold current densities under 200 A/cm[sup 2] and total differential efficiencies greater than 60% have been measured. DBR ring oscillators have demonstrated over 1 W of single frequency output power, 460 mW of spatially coherent, single frequency output power, and nearly circular diffraction limited output to 4 [times] I[sub th]. The performance potential of these devices is enormous, considering that the output apertures are nearly two orders of magnitude wider than conventional single mode sources which generate up to 0.2 W of coherent output.

  7. Thermo/opto/mechanical analysis of large apertures for exoplanet detection using Cielo

    NASA Astrophysics Data System (ADS)

    Jordan, Elizabeth O.; Chainyk, Mike; Habbal, Feras; Hoff, Claus; Levine, Marie; Moore, Greg

    2009-08-01

    The next generation of space telescopes will be designed to meet increasingly challenging science goals. The operating environment and required precision of these telescopes will make complete verification via ground tests impossible, and will place a greater reliance on numerical simulation. The current state of the art in thermal, mechanical and optical modeling involves three disparate computational models, several analysis codes and tools to transition results between these models. However, the active controls necessary to meet the next generation of requirements for space telescopes will require integrated thermal, structural, optical and controls analysis. To meet these challenges, JPL has developed Cielo, an in-house finite element tool capable of multi-physics simulations using a common finite element model, for thermal, structural and optical aberration analysis. In this paper, we will discuss the use of Cielo for analysis of a coronagraph and an occulter designed to observe Earth-like planets around nearby stars. We will compare thermal and structural results from Cielo with results from commercial off the shelf (COTS) tools to verify the new approach. We will perform variations of key parameters to demonstrate how margins and uncertainties can be quantified using the new approach.

  8. T/R module development for large aperture L-band phased array

    NASA Technical Reports Server (NTRS)

    Chamberlain, Neil; Andricos, Constantine; Kumley, Kendra; Berkun, Andrew; Hodges, Richard; Spitz, Suzanne

    2004-01-01

    This paper describes a transmit / receive (T/R) module for a large L-band space based radar active phased array being developed at JPL. Electrical performance and construction techniques are described, with emphasis on the former. The T/R modules have a bandwidth of more than 80 MHz centered at 1260MHz and support dual, switched polarizations. Phase and amplitude are controlled by a 6-bit phase shifter and a 6-bit attenuator, respectively. The transmitter power amplifier generates 2.4 W into a nominal 50 ohm load with 36% overall efficiency. The receiver noise figure is 4.4 dB including all front-end losses. The module weighs 32 g and has a footprint of 8 cm x 4.5 cm. Fourteen of these T/R modules were fabricated at the JPL Pick-and-Place Facility and were tested using a computer-controlled measurement facility developed at JPL. Calibrated performance of this set of T/R modules is presented and shows good agreement with design predictions.

  9. Quantitative comparison of terahertz emission from (100) InAs surfaces and a GaAs large-aperture photoconductive switch at high fluences.

    PubMed

    Reid, Matthew; Fedosejevs, Robert

    2005-01-01

    InAs has previously been reported to be an efficient emitter of terahertz radiation at low excitation fluences by use of femtosecond laser pulses. The scaling and saturation of terahertz emission from a (100) InAs surface as a function of excitation fluence is measured and quantitatively compared with the emission from a GaAs large-aperture photoconductive switch. We find that, although the instantaneous peak radiated terahertz field from (100) InAs exceeds the peak radiated signals from a GaAs large-aperture photoconductive switch biased at 1.6 kV/cm, the pulse duration is shorter. For the InAs source the total energy radiated is less than can be obtained from a GaAs large-aperture photoconductive switch. PMID:15662896

  10. Designing Mimics of Membrane Active Proteins

    PubMed Central

    Sgolastra, Federica; deRonde, Brittany M.; Sarapas, Joel M.; Som, Abhigyan; Tew, Gregory N.

    2014-01-01

    CONSPECTUS As a semi-permeable barrier that controls the flux of biomolecules in and out the cell, the plasma membrane is critical in cell function and survival. Many proteins interact with the plasma membrane and modulate its physiology. Within this large landscape of membrane-active molecules, researchers have focused significant attention on two specific classes of peptides, antimicrobial peptides (AMPs) and cell penetrating peptides (CPPs) because of their unique properties. In this account, we describe our efforts over the last decade to build and understand synthetic mimics of antimicrobial peptides (SMAMPs). These endeavors represent one specific example of a much larger effort to understand how synthetic molecules interact with and manipulate the plasma membrane. Using both defined molecular weight oligomers and easier to produce, but heterogeneous, polymers, it has been possible to generate scaffolds with biological potency superior to the natural analogs. In one case, a compound has progressed through a phase II clinical trial for pan)staph infections. Modern biophysical assays highlighted the interplay between the synthetic scaffold and lipid composition leading to negative Gaussian curvature, a requirement for both pore formation and endosomal escape. The complexity of this interplay between lipids, bilayer components, and the scaffolds remains to be better resolved, but significant new insight has been provided. It is worthwhile to consider the various aspects of permeation and how these are related to ‘pore formation.’ More recently, our efforts have expanded toward protein transduction domains, or cell penetrating peptide, mimics. The combination of unique molecular scaffolds and guanidinium) rich side chains has produced an array of polymers with robust transduction (and delivery) activity. Being a new area, the fundamental interactions between these new scaffolds and the plasma membrane are just beginning to be understood. Negative Gaussian

  11. Large aperture kinoform phase plates in fused silica for spatial beam smoothing on Nova and the Beamlet Lasers

    SciTech Connect

    Rushford, M.C.; Dixit, S.N.; Thomas, I.M.; Martin, A.M.; Perry, M.D.

    1997-03-01

    It is now widely recognized that spatial beam smoothing (homogenization) is essential in coupling the laser energy to the inertial confinement fusion (ICF) targets. For the indirect drive approach to ICF, it is desirable to distribute the laser energy into a uniformly speckled profile that has a flat-top super-Gaussian envelope (8th power or higher) and contains greater than 95% of the energy inside the super-Gaussian profile. Spatial smoothing is easily achieved by introducing a binary random phase plate (RPP) in the beam. This produces a homogenized far-field pattern which consists of an overall envelope function determined by the RPP element superimposed with a fine scale speckle pattern arising due to the interference among the various RPP elements. Although easy to fabricate and currently in routine use in many fusion laboratories, the binary RPPs do not meet the ICF requirements stated above since the far-field intensity profile is restricted to essentially an Airy function containing only 84% (an upper limit) of the energy inside the central spot. Approaches using lenslet arrays (refractive or diffractive) have limited use since they operate in the quasi-far-field and have a short depth of focus. The limitations of the RPPs can be overcome by relaxing the binary phase constraint. We have recently presented 5 continuously varying phase screens for tailoring the focal plane irradiance profiles. Called kinoform phase plates (KPPs), these phase screens offer complete flexibility in tailoring the focal plane envelope and, at the same time, increasing the energy efficiency inside the focal spot. In this paper we discuss the design and fabrication of such kinoform phase plates in fused silica for spatial beam smoothing on the Nova and the Beamlet lasers. Since the phase plates are used at the end of the laser chain, KPPs on Nova and Beamlet have to be fabricated on large aperture optics (65-cm diameter and 40-cm square substrates respectively). The following

  12. Lupus I Observations from the 2010 Flight of the Balloon-borne Large Aperture Submillimeter Telescope for Polarimetry

    NASA Astrophysics Data System (ADS)

    Matthews, Tristan G.; Ade, Peter A. R.; Angilè, Francesco E.; Benton, Steven J.; Chapin, Edward L.; Chapman, Nicholas L.; Devlin, Mark J.; Fissel, Laura M.; Fukui, Yasuo; Gandilo, Natalie N.; Gundersen, Joshua O.; Hargrave, Peter C.; Klein, Jeffrey; Korotkov, Andrei L.; Moncelsi, Lorenzo; Mroczkowski, Tony K.; Netterfield, Calvin B.; Novak, Giles; Nutter, David; Olmi, Luca; Pascale, Enzo; Poidevin, Frédérick; Savini, Giorgio; Scott, Douglas; Shariff, Jamil A.; Soler, Juan Diego; Tachihara, Kengo; Thomas, Nicholas E.; Truch, Matthew D. P.; Tucker, Carole E.; Tucker, Gregory S.; Ward-Thompson, Derek

    2014-04-01

    The Balloon-borne Large Aperture Submillimeter Telescope for Polarimetry (BLASTPol) was created by adding polarimetric capability to the BLAST experiment that was flown in 2003, 2005, and 2006. BLASTPol inherited BLAST's 1.8 m primary and its Herschel/SPIRE heritage focal plane that allows simultaneous observation at 250, 350, and 500 μm. We flew BLASTPol in 2010 and again in 2012. Both were long duration Antarctic flights. Here we present polarimetry of the nearby filamentary dark cloud Lupus I obtained during the 2010 flight. Despite limitations imposed by the effects of a damaged optical component, we were able to clearly detect submillimeter polarization on degree scales. We compare the resulting BLASTPol magnetic field map with a similar map made via optical polarimetry. (The optical data were published in 1998 by J. Rizzo and collaborators.) The two maps partially overlap and are reasonably consistent with one another. We compare these magnetic field maps to the orientations of filaments in Lupus I, and we find that the dominant filament in the cloud is approximately perpendicular to the large-scale field, while secondary filaments appear to run parallel to the magnetic fields in their vicinities. This is similar to what is observed in Serpens South via near-IR polarimetry, and consistent with what is seen in MHD simulations by F. Nakamura and Z. Li.

  13. Study on the Stressed Mirror Polishing with a Continuous Polishing Machine for Large Aperture Off-axis Aspheric Mirrors

    NASA Astrophysics Data System (ADS)

    Li, Xin-nan; Zhang, Hai-ying; Cui, Xiang-qun; Jiang, Zi-bo; Zheng, Yi; Liu, Xing-tao; Ni, Hou-kun

    2012-10-01

    A special stressed annular polishing technique is proposed to mill the off-axis aspheric sub-mirrors of a large segmented mirror with an annular polishing machine. Based on the basic principle of stressed annular polishing technique, a set of special stressing mechanisms are designed to convert milling the aspheric surfaces of sub-mirrors with different off-axis distances into milling the spherical surfaces with identical radii of curvature, so that they can be pol- ished simultaneously on a continuous polishing machine. It took about contin- uous 40 hours to polish a scaled-down mirror of the planning Chinese Future Giant Telescope (CFGT) using this technique. This mirror has the 330 mm di- ameter, 3.6 m off-axis distance, and the 21.6 m radius of curvature, and its max- imum asphericity is 16 micron. The experiment shows that this method has a high effciency, suits batch manufacturing, especially the batch manufacturing of aspheric sub-mirrors of the segmented primary mirror of an extremely large aperture telescope.

  14. Spaceborne Microwave Instrument for High Resolution Remote Sensing of the Earth's Surface Using a Large-Aperture Mesh Antenna

    NASA Technical Reports Server (NTRS)

    Njoku, E.; Wilson, W.; Yueh, S.; Freeland, R.; Helms, R.; Edelstein, W.; Sadowy, G.; Farra, D.; West, R.; Oxnevad, K.

    2001-01-01

    This report describes a two-year study of a large-aperture, lightweight, deployable mesh antenna system for radiometer and radar remote sensing of the Earth from space. The study focused specifically on an instrument to measure ocean salinity and Soil moisture. Measurements of ocean salinity and soil moisture are of critical . importance in improving knowledge and prediction of key ocean and land surface processes, but are not currently obtainable from space. A mission using this instrument would be the first demonstration of deployable mesh antenna technology for remote sensing and could lead to potential applications in other remote sensing disciplines that require high spatial resolution measurements. The study concept features a rotating 6-m-diameter deployable mesh antenna, with radiometer and radar sensors, to measure microwave emission and backscatter from the Earth's surface. The sensors operate at L and S bands, with multiple polarizations and a constant look angle, scanning across a wide swath. The study included detailed analyses of science requirements, reflector and feedhorn design and performance, microwave emissivity measurements of mesh samples, design and test of lightweight radar electronic., launch vehicle accommodations, rotational dynamics simulations, and an analysis of attitude control issues associated with the antenna and spacecraft, The goal of the study was to advance the technology readiness of the overall concept to a level appropriate for an Earth science emission.

  15. Cavity-excited Huygens' metasurface antennas for near-unity aperture illumination efficiency from arbitrarily large apertures.

    PubMed

    Epstein, Ariel; Wong, Joseph P S; Eleftheriades, George V

    2016-01-01

    One of the long-standing problems in antenna engineering is the realization of highly directive beams using low-profile devices. In this paper, we provide a solution to this problem by means of Huygens' metasurfaces (HMSs), based on the equivalence principle. This principle states that a given excitation can be transformed to a desirable aperture field by inducing suitable electric and (equivalent) magnetic surface currents. Building on this concept, we propose and demonstrate cavity-excited HMS antennas, where the single-source-fed cavity is designed to optimize aperture illumination, while the HMS facilitates the current distribution that ensures phase purity of aperture fields. The HMS breaks the coupling between the excitation and radiation spectra typical to standard partially reflecting surfaces, allowing tailoring of the aperture properties to produce a desirable radiation pattern, without incurring edge-taper losses. The proposed low-profile design yields near-unity aperture illumination efficiencies from arbitrarily large apertures, offering new capabilities for microwave, terahertz and optical radiators. PMID:26790605

  16. Cavity-excited Huygens' metasurface antennas for near-unity aperture illumination efficiency from arbitrarily large apertures

    NASA Astrophysics Data System (ADS)

    Epstein, Ariel; Wong, Joseph P. S.; Eleftheriades, George V.

    2016-01-01

    One of the long-standing problems in antenna engineering is the realization of highly directive beams using low-profile devices. In this paper, we provide a solution to this problem by means of Huygens' metasurfaces (HMSs), based on the equivalence principle. This principle states that a given excitation can be transformed to a desirable aperture field by inducing suitable electric and (equivalent) magnetic surface currents. Building on this concept, we propose and demonstrate cavity-excited HMS antennas, where the single-source-fed cavity is designed to optimize aperture illumination, while the HMS facilitates the current distribution that ensures phase purity of aperture fields. The HMS breaks the coupling between the excitation and radiation spectra typical to standard partially reflecting surfaces, allowing tailoring of the aperture properties to produce a desirable radiation pattern, without incurring edge-taper losses. The proposed low-profile design yields near-unity aperture illumination efficiencies from arbitrarily large apertures, offering new capabilities for microwave, terahertz and optical radiators.

  17. Performance of large aperture tapered fiber phase conjugate mirror with high pulse energy and 1-kHz repetition rate.

    PubMed

    Zhao, Zhigang; Dong, Yantao; Pan, Sunqiang; Liu, Chong; Chen, Jun; Tong, Lixin; Gao, Qingsong; Tang, Chun

    2012-01-16

    A large aperture fused silica tapered fiber phase conjugate mirror is presented with a maximum 70% stimulated Brillouin scattering (SBS) reflectivity, which is obtained with 1 kHz repetition rate, 15 ns pulse width and 38 mJ input pulse energy. To the best of our knowledge, this is the highest SBS reflectivity ever reported by using optical fiber as a phase conjugate mirror for such high pulse repetition rate (1 kHz) and several tens of millijoule (mJ) input pulse energy. The influences of fiber end surface quality and pump pulse widths on SBS reflectivity are investigated experimentally. The results show that finer fiber end surface quality and longer input pulse widths are preferred for obtaining higher SBS reflectivity with higher input pulse energy. Double passing amplification experiments are also performed. 52 mJ pulse energy is achieved at 1 kHz repetition rate, with a reflected SBS pulse width of 1.5 ns and a M(2) factor of 2.3. The corresponding peak power reaches 34.6 MW. Obvious beam quality improvement is observed. PMID:22274534

  18. Cavity-excited Huygens' metasurface antennas for near-unity aperture illumination efficiency from arbitrarily large apertures

    PubMed Central

    Epstein, Ariel; Wong, Joseph P. S.; Eleftheriades, George V.

    2016-01-01

    One of the long-standing problems in antenna engineering is the realization of highly directive beams using low-profile devices. In this paper, we provide a solution to this problem by means of Huygens' metasurfaces (HMSs), based on the equivalence principle. This principle states that a given excitation can be transformed to a desirable aperture field by inducing suitable electric and (equivalent) magnetic surface currents. Building on this concept, we propose and demonstrate cavity-excited HMS antennas, where the single-source-fed cavity is designed to optimize aperture illumination, while the HMS facilitates the current distribution that ensures phase purity of aperture fields. The HMS breaks the coupling between the excitation and radiation spectra typical to standard partially reflecting surfaces, allowing tailoring of the aperture properties to produce a desirable radiation pattern, without incurring edge-taper losses. The proposed low-profile design yields near-unity aperture illumination efficiencies from arbitrarily large apertures, offering new capabilities for microwave, terahertz and optical radiators. PMID:26790605

  19. Low stress ion-assisted coatings on fused silica substrates for large aperture laser pulse compression gratings

    NASA Astrophysics Data System (ADS)

    Smith, Douglas J.; McCullough, Mike; Smith, Claire; Mikami, Takuya; Jitsuno, Takahisa

    2008-10-01

    Large aperture laser pulse compressor designs use several diffraction gratings in series and sometimes tiled together to compress an amplified 1 to 10 ns pulse to 0.1 to 10 ps. The wavefront of the compressed pulse must be well controlled to allow focusing to a small spot on a target. Traditionally, multilayer dielectric gratings (MLDG) have been fabricated onto high thermal expansion substrates such as BK7 glass to prevent crazing and excessive bending due to tensile coating stress when operated in high vacuum. However, the high CTE of the BK7 can cause wavefront distortion and changes in the period of the grating. This work uses ion-assisted deposition of HfO2/SiO2 films to increase the compressive stress in MLD layers to allow use of silica substrates in the compressor vacuum environment. Stress, coating uniformity, and damage results are reported. The process was scaled to full size (91cm × 42cm) MLD gratings for use in the Osaka University LFEX laser system. Diffracted wavefront results from the full scale gratings is presented.

  20. Seasonal variability of turbulent fluxes over a vegetated subtropical coastal wetland measured by large aperture scintillometry and eddy covariance

    NASA Astrophysics Data System (ADS)

    Guyot, Adrien; Gray, Michael; Riesenkamp, Michiel; Lockington, David; McGowan, Hamish

    2016-04-01

    Subtropical coastal wetlands are particularly susceptible to the impacts of climate variability: their recharge rates strongly depend on rainfall, and the occurrence of prolonged droughts or wet periods have direct consequences for wetland health and bio-diversity. There is therefore a need to close the water budget of these ecosystems and this requires the quantification of rates of evaporation/evapotranspiration. However, few studies have documented land-atmosphere exchanges over wetlands for which water level varies considerably during a typical annual cycle. Here, we present a year of turbulent flux observations over a wetland on the subtropical coast of eastern Australia. Large Aperture Scintillometry and Eddy Covariance are used to derive sensible heat fluxes. Latent heat fluxes are also derived through an energy balance for both instruments' observations and also directly through Eddy Covariance. Careful sensitivity analysis of the instrumental footprints, seasonal variations of land surface parameters such as roughness length and displacement height are examined and subsequent uncertainties in the derived turbulent fluxes are discussed. Finally we show how these observations can also help better understand hydrological processes at the catchment scale.

  1. LUPUS I observations from the 2010 flight of the Balloon-borne large aperture submillimeter telescope for polarimetry

    SciTech Connect

    Matthews, Tristan G.; Chapman, Nicholas L.; Novak, Giles; Ade, Peter A. R.; Hargrave, Peter C.; Nutter, David; Angilè, Francesco E.; Devlin, Mark J.; Klein, Jeffrey; Benton, Steven J.; Fissel, Laura M.; Gandilo, Natalie N.; Netterfield, Calvin B.; Chapin, Edward L.; Fukui, Yasuo; Gundersen, Joshua O.; Korotkov, Andrei L.; Moncelsi, Lorenzo; Mroczkowski, Tony K.; Olmi, Luca; and others

    2014-04-01

    The Balloon-borne Large Aperture Submillimeter Telescope for Polarimetry (BLASTPol) was created by adding polarimetric capability to the BLAST experiment that was flown in 2003, 2005, and 2006. BLASTPol inherited BLAST's 1.8 m primary and its Herschel/SPIRE heritage focal plane that allows simultaneous observation at 250, 350, and 500 μm. We flew BLASTPol in 2010 and again in 2012. Both were long duration Antarctic flights. Here we present polarimetry of the nearby filamentary dark cloud Lupus I obtained during the 2010 flight. Despite limitations imposed by the effects of a damaged optical component, we were able to clearly detect submillimeter polarization on degree scales. We compare the resulting BLASTPol magnetic field map with a similar map made via optical polarimetry. (The optical data were published in 1998 by J. Rizzo and collaborators.) The two maps partially overlap and are reasonably consistent with one another. We compare these magnetic field maps to the orientations of filaments in Lupus I, and we find that the dominant filament in the cloud is approximately perpendicular to the large-scale field, while secondary filaments appear to run parallel to the magnetic fields in their vicinities. This is similar to what is observed in Serpens South via near-IR polarimetry, and consistent with what is seen in MHD simulations by F. Nakamura and Z. Li.

  2. A fundamental mode Nd:GdVO4 laser pumped by a large aperture 808 nm VCSEL

    NASA Astrophysics Data System (ADS)

    Hao, Y. Q.; Ma, J. L.; Yan, C. L.; Liu, G. J.; Ma, X. H.; Gong, J. F.; Feng, Y.; Wei, Z. P.; Wang, Y. X.; Zhao, Y. J.

    2013-05-01

    A fundamental mode Nd:GdVO4 laser pumped by a vertical cavity surface emitting laser (VCSEL) is experimentally demonstrated. The VCSEL has a circular output-beam which makes it easier for it to be directly coupled to a Nd:GdVO4 microcrystal. In our research, a large aperture 808 nm VCSEL, with a multi-ring-shaped aperture (MRSA) and an almost Gaussian-shaped far-field profile, is used as the pumping source. Experimental results for the Nd:GdVO4 laser pumped by the VCSEL are presented. The maximum output peak power of 0.754 W is obtained under a pump peak power of 1.3 W, and the corresponding opto-optic conversion efficiency is 58.1%. The average slope efficiency is 65.8% from the threshold pump power of 0.2 W to the pump power of 1.3 W. The laser beam quality factors are measured to be {M}x2=1.2 0 and {M}y2=1.1 5.

  3. MRF Applications: On the Road to Making Large-Aperture Ultraviolet Laser Resistant Continuous Phase Plates for High-Power Lasers

    SciTech Connect

    Menapace, J A; Davis, P J; Steele, W A; Hachkowski, M R; Nelson, A; Xin, K

    2006-10-26

    Over the past two years we have developed MRF tools and procedures to manufacture large-aperture (430 X 430 mm) continuous phase plates (CPPs) that are capable of operating in the infrared portion (1053 nm) of high-power laser systems. This is accomplished by polishing prescribed patterns of continuously varying topographical features onto finished plano optics using MRF imprinting techniques. We have been successful in making, testing, and using large-aperture CPPs whose topography possesses spatial periods as low as 4 mm and surface peak-to-valleys as high as 8.6 {micro}m. Combining this application of MRF technology with advanced MRF finishing techniques that focus on ultraviolet laser damage resistance makes it potentially feasible to manufacture large-aperture CPPs that can operate in the ultraviolet (351 nm) without sustaining laser-induced damage. In this paper, we will discuss the CPP manufacturing process and the results of 351-nm/3-nsec equivalent laser performance experiments conducted on large-aperture CPPs manufactured using advanced MRF protocols.

  4. Analysis of fratricide effect observed with GeMS and its relevance for large aperture astronomical telescopes

    NASA Astrophysics Data System (ADS)

    Otarola, Angel; Neichel, Benoit; Wang, Lianqi; Boyer, Corinne; Ellerbroek, Brent; Rigaut, François

    2013-12-01

    Large aperture ground-based telescopes require Adaptive Optics (AO) to correct for the distortions induced by atmospheric turbulence and achieve diffraction limited imaging quality. These AO systems rely on Natural and Laser Guide Stars (NGS and LGS) to provide the information required to measure the wavefront from the astronomical sources under observation. In particular one such LGS method consists in creating an artificial star by means of fluorescence of the sodium atoms at the altitude of the Earth's mesosphere. This is achieved by propagating one or more lasers, at the wavelength of the Na D2a resonance, from the telescope up to the mesosphere. Lasers can be launched from either behind the secondary mirror or from the perimeter of the main aperture. The so-called central- and side-launch systems, respectively. The central-launch system, while helpful to reduce the LGS spot elongation, introduces the so-called "fratricide" effect. This consists of an increase in the photon-noise in the AO Wave Front Sensors (WFS) sub-apertures, with photons that are the result of laser photons back-scattering from atmospheric molecules (Rayleigh scattering) and atmospheric aerosols (dust and/or cirrus clouds ice particles). This affects the performance of the algorithms intended to compute the LGS centroids and subsequently compute and correct the turbulence-induced wavefront distortions. In the frame of the Thirty Meter Telescope (TMT) project and using actual LGS WFS data obtained with the Gemini Multi-Conjugate Adaptive Optics System (Gemini MCAO a.k.a. GeMS), we show results from an analysis of the temporal variability of the observed fratricide effect, as well as comparison of the absolute magnitude of fratricide photon-flux level with simulations using models that account for molecular (Rayleigh) scattering and photons backscattered from cirrus clouds.

  5. Modeling the global micrometeor input function in the upper atmosphere observed by high power and large aperture radars

    NASA Astrophysics Data System (ADS)

    Janches, Diego; Heinselman, Craig J.; Chau, Jorge L.; Chandran, Amal; Woodman, Ronald

    2006-07-01

    We report initial results of an effort to model the diurnal and seasonal variability of the meteor rate detected by high power and large aperture (HPLA) radars. The model uses Monte Carlo simulation techniques and at present assumes that most of the detected particles originate from three radiant distributions with the most dominant concentrated around the Earth's apex. The other two sources are centered 80° in ecliptic longitude to each side of the apex and are commonly known as helion and antihelion. To reproduce the measurements, the apex source flux was set to provide ˜70% of the total number of particles while the other ˜30% is provided by the combined contribution of the two remaining sources. The results of the model are in excellent agreement with observed diurnal curves obtained at different seasons and locations using the 430 MHz Arecibo radar in Puerto Rico, the 50 MHz Jicamarca radar in Perú, and the 1.29 GHz Sondrestrom radar in Greenland. To obtain agreement with the observed diurnal and seasonal variability of the meteor rate, an empirical atmospheric filtering effect was introduced in the simulation which prevents meteors with low-elevation radiants (≤20°) from being detected by the radars at mesospheric altitudes. The filtering effect is probably produced by a combination of factors related to the interaction of the meteor with the air molecules such as electron production and/or the ablation at higher altitudes. On the basis of these results we calculate the micrometeor global, diurnal, and seasonal input in the upper atmosphere.

  6. Estimation of turbulent sensible heat and momentum fluxes over a heterogeneous urban area using a large aperture scintillometer

    NASA Astrophysics Data System (ADS)

    Lee, Sang-Hyun; Lee, Jun-Ho; Kim, Bo-Young

    2015-08-01

    The accurate determination of surface-layer turbulent fluxes over urban areas is critical to understanding urban boundary layer (UBL) evolution. In this study, a remote-sensing technique using a large aperture scintillometer (LAS) was investigated to estimate surface-layer turbulent fluxes over a highly heterogeneous urban area. The LAS system, with an optical path length of 2.1 km, was deployed in an urban area characterized by a complicated land-use mix (residential houses, water body, bare ground, etc.). The turbulent sensible heat ( Q H) and momentum fluxes (τ) were estimated from the scintillation measurements obtained from the LAS system during the cold season. Three-dimensional LAS footprint modeling was introduced to identify the source areas ("footprint") of the estimated turbulent fluxes. The analysis results showed that the LAS-derived turbulent fluxes for the highly heterogeneous urban area revealed reasonable temporal variation during daytime on clear days, in comparison to the land-surface process-resolving numerical modeling. A series of sensitivity tests indicated that the overall uncertainty in the LAS-derived daytime Q H was within 20%-30% in terms of the influence of input parameters and the nondimensional similarity function for the temperature structure function parameter, while the estimation errors in τ were less sensitive to the factors of influence, except aerodynamic roughness length. The 3D LAS footprint modeling characterized the source areas of the LAS-derived turbulent fluxes in the heterogeneous urban area, revealing that the representative spatial scales of the LAS system deployed with the 2.1 km optical path distance ranged from 0.2 to 2 km2 (a "micro- a scale"), depending on local meteorological conditions.

  7. Determining meteoroid bulk densities using a plasma scattering model with high-power large-aperture radar data

    NASA Astrophysics Data System (ADS)

    Close, Sigrid; Volz, Ryan; Loveland, Rohan; Macdonell, Alex; Colestock, Patrick; Linscott, Ivan; Oppenheim, Meers

    2012-09-01

    We present an improved technique for calculating bulk densities of low-mass (<1 g) meteoroids using a scattering model applied to the high-density plasma formed around the meteoroid as it enters Earth’s atmosphere. These plasmas, referred to as head echoes, travel at or near the speed of the meteoroid, thereby allowing the determination of the ballistic coefficient (mass divided by physical cross-section), which depends upon speed and deceleration. Concurrently, we apply a scattering model to the returned signal strength of the head echo in order to correlate radar-cross-section (RCS) to plasma density and meteoroid mass. In this way, we can uniquely solve for the meteoroid mass, radius and bulk density independently. We have applied this new technique to head echo data collected in 2007 and 2008 simultaneously at VHF (160 MHz) and UHF (422 MHz) at ALTAIR, which is a high-power large-aperture radar located on the Kwajalein Atoll. These data include approximately 20,000 detections with dual-frequency, dual-polarization, and monopulse (i.e. angle) returns. From 2000 detections with the smallest monopulse errors, we find a mean meteoroid bulk density of 0.9 g/cm3 with observations spanning almost three orders of magnitude from 0.01 g/cm3 to 8 g/cm3. Our results show a clear dependence between meteoroid bulk density and altitude of head echo formation, as well as dependence between meteoroid bulk density and 3D speed. The highest bulk densities are detected at the lowest altitudes and lowest speeds. Additionally, we stipulate that the approximations used to derive the ballistic parameter, in addition to neglecting fragmentation, suggest that the traditional ballistic parameter must be used with caution when determining meteoroid parameters.

  8. Large-Aperture [O I] 6300 A Photometry of Comet Hale-Bopp: Implications for the Photochemistry of OH

    NASA Technical Reports Server (NTRS)

    Morgenthaler, Jeffrey P.; Harris, Walter M.; Scherb, Frank; Anderson, Christopher M.; Oliversen, Ronald J.; Doane, Nathaniel E.; Combi, Michael R.; Marconi, Maximus L.; Smyth, William H.

    2001-01-01

    Large-aperture photometric observations of comet Hale-Bopp (C/1995 O1) in the forbidden red line of neutral oxygen ([O I] 6300 angstroms) with the 150 mm dual-etalon Fabry-Perot spectrometer that comprises the Wisconsin H-alpha Mapper and a 50 mm dual-etalon Fabry-Perot spectrometer at the McMath-Pierce main telescope from 1997 late February to mid April yield a total metastable O((sup 1)D) production rate of (2.3-5.9) x 10(exp 30)/s. Applying the standard H2O and OH photodissociation branching ratios, we derive a water production rate, Q(H2O), of (2.6-6.1) x 10(exp 31)/s, which disagrees with Q(H2O = 1x10(exp 31)/s determined by independent H2O, OH, and H measurements. Furthermore, our own [O I] 6300 observations of the inner coma (< 30,000 km) using the 3.5 m Wisconsin-Indiana-Yale-NOAO telescope Hydra and Densepak multi-object spectrographs yield Q(H2O) = 1 x 10(exp 31)/s. Using our [O I] 6300 data, which cover spatial scales ranging from 2,000 to 1x10(exp 6) km, and a complementary set of wide-field ground-based OH images, we can constrain the sources of the apparent excess O((sup 1)D) emission to the outer coma, where photodissociation of OH is assumed to be the dominant O((sup 1)D) production mechanism. From production rates of other oxygen-bearing volatiles (e.g., CO and CO2), we can account for at most 30% of the observed excess O((sup 1)D) emission. Since even less O((sup 1)D) should be coming from other sources (e.g., electron excitation of neutral O and distributed nonnuclear sources of H2O), we hypothesize that the bulk of the excess O((sup 1)D) is likely coming from photodissociating OH. Using the experimental OH photo-dissociation cross section of Nee and Lee at Ly-alpha as a guide in modifying the theoretical OH cross sections of van Dishoeck and Dalgarno, we can account for approximately 60% of the observed O((sup 1)D) excess without requiring major modifications to the other OH branching ratios or the total OH photodissociation lifetime.

  9. Water permeation through Nafion membranes: the role of water activity.

    PubMed

    Majsztrik, Paul; Bocarsly, Andrew; Benziger, Jay

    2008-12-25

    The permeation of water through 1100 equivalent weight Nation membranes has been measured for film thicknesses of 51-254 microm, temperatures of 30-80 degrees C, and water activities (a(w)) from 0.3 to 1 (liquid water). Water permeation coefficients increased with water content in Nafion. For feed side water activity in the range 0 < a(w) < 0.8, permeation coefficients increased linearly with water activity and scaled inversely with membrane thickness. The permeation coefficients were independent of membrane thickness when the feed side of the membrane was in contact with liquid water (a(w) = 1). The permeation coefficient for a 127 microm thick membrane increased by a factor of 10 between contacting the feed side of the membrane to water vapor (a(w) = 0.9) compared to liquid water (a(w) = 1). Water permeation couples interfacial transport across the fluid membrane interface with water transport through the hydrophilic phase of Nafion. At low water activity the hydrophilic volume fraction is small and permeation is limited by water diffusion. The volume fraction of the hydrophilic phase increases with water activity, increasing water transport. As a(w) --> 1, the effective transport rate increased by almost an order of magnitude, resulting in a change of the limiting transport resistance from water permeation across the membrane to interfacial mass transport at the gas/membrane interface. PMID:19053672

  10. The Structural Basis of Cholesterol Activity in Membranes

    SciTech Connect

    Olsen, Brett N.; Bielska, Agata; Lee, Tiffany; Daily, Michael D.; Covey, Douglas F.; Schlesinger, Paul H.; Baker, Nathan A.; Ory, Daniel S.

    2013-10-15

    Although the majority of free cellular cholesterol is present in the plasma membrane, cholesterol homeostasis is principally regulated through sterol-sensing proteins that reside in the cholesterol-poor endoplasmic reticulum (ER). In response to acute cholesterol loading or depletion, there is rapid equilibration between the ER and plasma membrane cholesterol pools, suggesting a biophysical model in which the availability of plasma membrane cholesterol for trafficking to internal membranes modulates ER membrane behavior. Previous studies have predominantly examined cholesterol availability in terms of binding to extramembrane acceptors, but have provided limited insight into the structural changes underlying cholesterol activation. In this study, we use both molecular dynamics simulations and experimental membrane systems to examine the behavior of cholesterol in membrane bilayers. We find that cholesterol depth within the bilayer provides a reasonable structural metric for cholesterol availability and that this is correlated with cholesterol-acceptor binding. Further, the distribution of cholesterol availability in our simulations is continuous rather than divided into distinct available and unavailable pools. This data provide support for a revised cholesterol activation model in which activation is driven not by saturation of membrane-cholesterol interactions but rather by bulk membrane remodeling that reduces membrane-cholesterol affinity.

  11. Development of active-transport membrane devices

    SciTech Connect

    Laciak, D.V.

    1994-07-01

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

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

    DOEpatents

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

    2012-09-24

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

  13. Nonequilibrium fluctuations, traveling waves, and instabilities in active membranes.

    PubMed

    Ramaswamy, S; Toner, J; Prost, J

    2000-04-10

    The stability of a flexible fluid membrane containing a distribution of mobile, active proteins (e.g., proton pumps) is shown to depend on the structure and functional asymmetry of the proteins. A stable active membrane is in a nonequilibrium steady state with height fluctuations whose statistical properties are governed by the protein activity. Disturbances are predicted to travel as waves at sufficiently long wavelength, with speed set by the normal velocity of the pumps. The unstable case involves a spontaneous, pump-driven undulation of the membrane, with clumping of the proteins in regions of high activity. PMID:11019123

  14. Membrane stretching triggers mechanosensitive Ca2+ channel activation in Chara.

    PubMed

    Kaneko, Toshiyuki; Takahashi, Naoya; Kikuyama, Munehiro

    2009-03-01

    In order to confirm that mechanosensitive Ca(2+) channels are activated by membrane stretching, we stretched or compressed the plasma membrane of Chara by applying osmotic shrinkage or swelling of the cell by varying the osmotic potential of the bathing medium. Aequorin studies revealed that treatments causing membrane stretching induced a transient but large increase in cytoplasmic concentration of Ca(2+) (Delta[Ca(2+)](c)). However, the observed Delta[Ca(2+)](c) decreased during the treatments, resulting in membrane compression. A second experiment was carried out to study the relationship between changes in membrane potential (DeltaE(m)) and stretching or compression of the plasma membrane. Significant DeltaE(m) values, often accompanied by an action potential, were observed during the initial exchange of the bathing medium from a hypotonic medium to a hypertonic one (plasmolysis). DeltaE(m) appears to be triggered by a partial stretching of the membrane as it was peeled from the cell wall. After plasmolysis, other exchanges from hypertonic to hypotonic media, with their accompanying membrane stretching, always induced large DeltaE(m) values and were often accompanied by an action potential. By contrast, action potentials were scarcely observed during other exchanges from hypotonic to hypertonic solutions (=membrane compression). Thus, we concluded that activation of the mechanosensitive channels is triggered by membrane stretching in Chara. PMID:19234734

  15. Functional Implications of Plasma Membrane Condensation for T Cell Activation

    PubMed Central

    Quinn, Carmel M.; Engelhardt, Karin; Williamson, David; Grewal, Thomas; Jessup, Wendy; Harder, Thomas; Gaus, Katharina

    2008-01-01

    The T lymphocyte plasma membrane condenses at the site of activation but the functional significance of this receptor-mediated membrane reorganization is not yet known. Here we demonstrate that membrane condensation at the T cell activation sites can be inhibited by incorporation of the oxysterol 7-ketocholesterol (7KC), which is known to prevent the formation of raft-like liquid-ordered domains in model membranes. We enriched T cells with 7KC, or cholesterol as control, to assess the importance of membrane condensation for T cell activation. Upon 7KC treatment, T cell antigen receptor (TCR) triggered calcium fluxes and early tyrosine phosphorylation events appear unaltered. However, signaling complexes form less efficiently on the cell surface, fewer phosphorylated signaling proteins are retained in the plasma membrane and actin restructuring at activation sites is impaired in 7KC-enriched cells resulting in compromised downstream activation responses. Our data emphasizes lipids as an important medium for the organization at T cell activation sites and strongly indicates that membrane condensation is an important element of the T cell activation process. PMID:18509459

  16. A common landscape for membrane-active peptides

    PubMed Central

    Last, Nicholas B; Schlamadinger, Diana E; Miranker, Andrew D

    2013-01-01

    Three families of membrane-active peptides are commonly found in nature and are classified according to their initial apparent activity. Antimicrobial peptides are ancient components of the innate immune system and typically act by disruption of microbial membranes leading to cell death. Amyloid peptides contribute to the pathology of diverse diseases from Alzheimer's to type II diabetes. Preamyloid states of these peptides can act as toxins by binding to and permeabilizing cellular membranes. Cell-penetrating peptides are natural or engineered short sequences that can spontaneously translocate across a membrane. Despite these differences in classification, many similarities in sequence, structure, and activity suggest that peptides from all three classes act through a small, common set of physical principles. Namely, these peptides alter the Brownian properties of phospholipid bilayers, enhancing the sampling of intrinsic fluctuations that include membrane defects. A complete energy landscape for such systems can be described by the innate membrane properties, differential partition, and the associated kinetics of peptides dividing between surface and defect regions of the bilayer. The goal of this review is to argue that the activities of these membrane-active families of peptides simply represent different facets of what is a shared energy landscape. PMID:23649542

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

    PubMed

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

    2016-05-01

    This paper focuses on automatic fuzzy clustering problem and proposes a novel automatic fuzzy clustering method that employs an extended membrane system with active membranes that has been designed as its computing framework. The extended membrane system has a dynamic membrane structure; since membranes can evolve, it is particularly suitable for processing the automatic fuzzy clustering problem. A modification of a differential evolution (DE) mechanism was developed as evolution rules for objects according to membrane structure and object communication mechanisms. Under the control of both the object's evolution-communication mechanism and the membrane evolution mechanism, the extended membrane system can effectively determine the most appropriate number of clusters as well as the corresponding optimal cluster centers. The proposed method was evaluated over 13 benchmark problems and was compared with four state-of-the-art automatic clustering methods, two recently developed clustering methods and six classification techniques. The comparison results demonstrate the superiority of the proposed method in terms of effectiveness and robustness. PMID:26790484

  18. Structural Model of Active Bax at the Membrane

    PubMed Central

    Bleicken, Stephanie; Jeschke, Gunnar; Stegmueller, Carolin; Salvador-Gallego, Raquel; García-Sáez, Ana J.; Bordignon, Enrica

    2016-01-01

    Bax plays a central role in the mitochondrial pathway of apoptosis. Upon activation, cytosolic Bax monomers oligomerize on the surface of mitochondria and change conformation concertedly to punch holes into the outer membrane. The subsequent release of cytochrome c initiates cell death. However, the structure of membrane-inserted Bax and its mechanism of action remain largely unknown. Here, we propose a 3D model of active Bax at the membrane based on double electron-electron resonance (DEER) spectroscopy in liposomes and isolated mitochondria. We show that active Bax is organized at the membrane as assemblies of dimers. In addition to a stable dimerization domain, each monomer contains a more flexible piercing domain involved in interdimer interactions and pore formation. The most important structural change during Bax activation is the opening of the hairpin formed by helices 5 and 6, which adopts a clamp-like conformation central to the mechanism of mitochondrial permeabilization. PMID:25458844

  19. Technology development for the Advanced Technology Large Aperture Space Telescope (ATLAST) as a candidate large UV-Optical-Infrared (LUVOIR) surveyor

    NASA Astrophysics Data System (ADS)

    Bolcar, Matthew R.; Balasubramanian, Kunjithapatham; Clampin, Mark; Crooke, Julie; Feinberg, Lee; Postman, Marc; Quijada, Manuel; Rauscher, Bernard; Redding, David; Rioux, Norman; Shaklan, Stuart; Stahl, H. Philip; Stahle, Carl; Thronson, Harley

    2015-09-01

    The Advanced Technology Large Aperture Space Telescope (ATLAST) team has identified five key technologies to enable candidate architectures for the future large-aperture ultraviolet/optical/infrared (LUVOIR) space observatory envisioned by the NASA Astrophysics 30-year roadmap, Enduring Quests, Daring Visions. The science goals of ATLAST address a broad range of astrophysical questions from early galaxy and star formation to the processes that contributed to the formation of life on Earth, combining general astrophysics with direct-imaging and spectroscopy of habitable exoplanets. The key technologies are: internal coronagraphs, starshades (or external occulters), ultra-stable large-aperture telescopes, detectors, and mirror coatings. Selected technology performance goals include: 1x10-10 raw contrast at an inner working angle of 35 milli-arcseconds, wavefront error stability on the order of 10 pm RMS per wavefront control step, autonomous on-board sensing and control, and zero-read-noise single-photon detectors spanning the exoplanet science bandpass between 400 nm and 1.8 μm. Development of these technologies will provide significant advances over current and planned observatories in terms of sensitivity, angular resolution, stability, and high-contrast imaging. The science goals of ATLAST are presented and flowed down to top-level telescope and instrument performance requirements in the context of a reference architecture: a 10-meter-class, segmented aperture telescope operating at room temperature (~290 K) at the sun-Earth Lagrange-2 point. For each technology area, we define best estimates of required capabilities, current state-of-the-art performance, and current Technology Readiness Level (TRL) - thus identifying the current technology gap. We report on current, planned, or recommended efforts to develop each technology to TRL 5.

  20. Technology Development for the Advanced Technology Large Aperture Space Telescope (ATLAST) as a Candidate Large UV-Optical-Infrared (LUVOIR) Surveyor

    NASA Technical Reports Server (NTRS)

    Bolcar, Matthew R.; Balasubramanian, Kunjithapatha; Clampin, Mark; Crooke, Julie; Feinberg, Lee; Postman, Marc; Quijada, Manuel; Rauscher, Bernard; Redding, David; Rioux, Norman; Shaklan, Stuart; Stahl, H. Philip; Stahle, Carl; Thronson, Harley

    2015-01-01

    The Advanced Technology Large Aperture Space Telescope (ATLAST) team has identified five key technologies to enable candidate architectures for the future large-aperture ultraviolet/optical/infrared (LUVOIR) space observatory envisioned by the NASA Astrophysics 30-year roadmap, Enduring Quests, Daring Visions. The science goals of ATLAST address a broad range of astrophysical questions from early galaxy and star formation to the processes that contributed to the formation of life on Earth, combining general astrophysics with direct-imaging and spectroscopy of habitable exoplanets. The key technologies are: internal coronagraphs, starshades (or external occulters), ultra-stable large-aperture telescopes, detectors, and mirror coatings. Selected technology performance goals include: 1x10?10 raw contrast at an inner working angle of 35 milli-arcseconds, wavefront error stability on the order of 10 pm RMS per wavefront control step, autonomous on-board sensing & control, and zero-read-noise single-photon detectors spanning the exoplanet science bandpass between 400 nm and 1.8 µm. Development of these technologies will provide significant advances over current and planned observatories in terms of sensitivity, angular resolution, stability, and high-contrast imaging. The science goals of ATLAST are presented and flowed down to top-level telescope and instrument performance requirements in the context of a reference architecture: a 10-meter-class, segmented aperture telescope operating at room temperature (290 K) at the sun-Earth Lagrange-2 point. For each technology area, we define best estimates of required capabilities, current state-of-the-art performance, and current Technology Readiness Level (TRL) - thus identifying the current technology gap. We report on current, planned, or recommended efforts to develop each technology to TRL 5.

  1. Comparison of CNES spherical and NASA hemispherical large aperture integrating sources. I - Using a laboratory transfer spectroradiometer. II - Using the SPOT-2 satellite instruments

    NASA Technical Reports Server (NTRS)

    Guenther, B.; Mclean, J.; Leroy, M.; Henry, P.

    1990-01-01

    CNES spherical and NASA hemispherical large aperture calibration sources are examined using a laboratory transfer spectroradiometer and SPOT-2 instruments. The sources, collected at Matra in France during October 1987, are compared in terms of absolute calibration, linearity, and uniformity. The laboratory transfer spectroradiometer data reveal that the calibration results correspond to within about 7 percent absolute accuracy level and the linearity of the CNES source with lamp level is good. It is observed using the satellite data that both sources have an excellent uniformity over a 4 deg field of view.

  2. Plane-polar Fresnel and far-field computations using the Fresnel-Wilcox and Jacobi-Bessel expansions. [for large aperture antennas

    NASA Technical Reports Server (NTRS)

    Rahmat-Samii, Y.; Galindo-Israel, V.

    1981-01-01

    It is pointed out that the computation of the Fresnel fields for large aperture antennas is significant for many applications. The present investigation is concerned with an approach for the effective utilization of the coefficients of the Jacobi-Bessel series for the far-field to obtain an analytically continuous representation of the antenna field which is valid from the Fresnel region into the far field. Attention is given to exact formulations and closed form solutions, Fresnel and Fresnel small angle approximations, aspects of field expansion, the accuracy of the Fresnel and Fresnel small angle approximations, and the Jacobi-Bessel expansion applied to the Fresnel small angle approximation.

  3. Ammonia-Activated Mesoporous Carbon Membranes for Gas Separations

    SciTech Connect

    Mahurin, Shannon Mark; Lee, Jeseung; Wang, Xiqing; Dai, Sheng

    2011-01-01

    Porous carbon membranes, which generally show improved chemical and thermal stability compared to polymer membranes, have been used in gas separations for many years. In this work, we show that the post-synthesis ammonia treatment of porous carbon at elevated temperature can improve the permeance and selectivity of these membranes for the separation of carbon dioxide and hydrocarbons from permanent gases. Hierarchically structured porous carbon membranes were exposed to ammonia gas at temperatures ranging from 850 C to 950 C for up to 10 min and the N{sub 2}, CO{sub 2}, and C{sub 3}H{sub 6} permeances were measured for these different membranes. Higher treatment temperatures and longer exposure times resulted in higher gas permeance values. In addition, CO{sub 2}/N{sub 2} and C{sub 3}H{sub 6}/N{sub 2} selectivities increased by a factor of 2 as the treatment temperature and time increased up to a temperature and time of 900 C, 10 min. Higher temperatures showed increased permeance but decreased selectivity indicating excess pore activation. Nitrogen adsorption measurements show that the ammonia treatment increased the porosity of the membrane while elemental analysis revealed the presence of nitrogen-containing surface functionalities in the treated carbon membranes. Thus, ammonia treatment at high temperature provides a controlled method to introduce both added microporosity and surface functionality to enhance gas separations performance of porous carbon membranes.

  4. Bilayer Membrane Modulation of Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) Structure and Proteolytic Activity.

    PubMed

    Cerofolini, Linda; Amar, Sabrina; Lauer, Janelle L; Martelli, Tommaso; Fragai, Marco; Luchinat, Claudio; Fields, Gregg B

    2016-01-01

    Cell surface proteolysis is an integral yet poorly understood physiological process. The present study has examined how the pericellular collagenase membrane-type 1 matrix metalloproteinase (MT1-MMP) and membrane-mimicking environments interplay in substrate binding and processing. NMR derived structural models indicate that MT1-MMP transiently associates with bicelles and cells through distinct residues in blades III and IV of its hemopexin-like domain, while binding of collagen-like triple-helices occurs within blades I and II of this domain. Examination of simultaneous membrane interaction and triple-helix binding revealed a possible regulation of proteolysis due to steric effects of the membrane. At bicelle concentrations of 1%, enzymatic activity towards triple-helices was increased 1.5-fold. A single mutation in the putative membrane interaction region of MT1-MMP (Ser466Pro) resulted in lower enzyme activation by bicelles. An initial structural framework has thus been developed to define the role(s) of cell membranes in modulating proteolysis. PMID:27405411

  5. Bilayer Membrane Modulation of Membrane Type 1 Matrix Metalloproteinase (MT1-MMP) Structure and Proteolytic Activity

    PubMed Central

    Cerofolini, Linda; Amar, Sabrina; Lauer, Janelle L.; Martelli, Tommaso; Fragai, Marco; Luchinat, Claudio; Fields, Gregg B.

    2016-01-01

    Cell surface proteolysis is an integral yet poorly understood physiological process. The present study has examined how the pericellular collagenase membrane-type 1 matrix metalloproteinase (MT1-MMP) and membrane-mimicking environments interplay in substrate binding and processing. NMR derived structural models indicate that MT1-MMP transiently associates with bicelles and cells through distinct residues in blades III and IV of its hemopexin-like domain, while binding of collagen-like triple-helices occurs within blades I and II of this domain. Examination of simultaneous membrane interaction and triple-helix binding revealed a possible regulation of proteolysis due to steric effects of the membrane. At bicelle concentrations of 1%, enzymatic activity towards triple-helices was increased 1.5-fold. A single mutation in the putative membrane interaction region of MT1-MMP (Ser466Pro) resulted in lower enzyme activation by bicelles. An initial structural framework has thus been developed to define the role(s) of cell membranes in modulating proteolysis. PMID:27405411

  6. Modeling and vibration control of an active membrane mirror

    NASA Astrophysics Data System (ADS)

    Ruggiero, Eric J.; Inman, Daniel J.

    2009-09-01

    The future of space satellite technology lies in ultra-large mirrors and radar apertures for significant improvements in imaging and communication bandwidths. The availability of optical-quality membranes drives a parallel effort for structural models that can capture the dominant dynamics of large, ultra-flexible satellite payloads. Unfortunately, the inherent flexibility of membrane mirrors wreaks havoc with the payload's on-orbit stability and maneuverability. One possible means of controlling these undesirable dynamics is by embedding active piezoelectric ceramics near the boundary of the membrane mirror. In doing so, active feedback control can be used to eliminate detrimental vibration, perform static shape control, and evaluate the health of the structure. The overall motivation of the present work is to design a control system using distributed bimorph actuators to eliminate any detrimental vibration of the membrane mirror. As a basis for this study, a piezoceramic wafer was attached in a bimorph configuration near the boundary of a tensioned rectangular membrane sample. A finite element model of the system was developed to capture the relevant system dynamics from 0 to 300 Hz. The finite element model was compared against experimental results, and fair agreement found. Using the validated finite element models, structural control using linear quadratic regulator control techniques was then used to numerically demonstrate effective vibration control. Typical results show that less than 12 V of actuation voltage is required to eliminate detrimental vibration of the membrane samples in less than 15 ms. The functional gains of the active system are also derived and presented. These spatially descriptive control terms dictate favorable regions within the membrane domain for placing sensors and can be used as a design guideline for structural control applications. The results of the present work demonstrate that thin plate theory is an appropriate modeling

  7. Membrane Changes Associated with Platelet Activation

    PubMed Central

    George, James N.; Lyons, Roger M.; Morgan, Rebecca K.

    1980-01-01

    The effect of aggregation and secretion on membrane proteins was studied in washed human platelets. Reversible aggregation without secretion was stimulated by ADP and secretion without aggregation was stimulated by thrombin in the presence of EDTA. No loss of platelet surface glycoproteins occurred during reversible ADP-induced platelet aggregation, as measured by quantitative polyacrylamide gel electrophoresis analysis of platelets that were labeled with 125I-diazotized diiodosulfanilic acid (DD125ISA) before ADP stimulation. Also, no new proteins became exposed on the platelet surface after ADP aggregation, as determined by DD125ISA labeling after stimulation. Thrombin-induced platelet secretion also caused no loss of platelet surface glycoproteins. However, after platelet secretion two new proteins were labeled by DD125ISA: (a) actin and (b) the 149,000-mol wt glycoprotein (termed GP-G), which is contained in platelet granules and secreted in response to thrombin. The identity of DD125ISA-labeled actin was confirmed by four criteria: (a) comigration with actin in three different sodium dodecyl sulfate-polyacrylamide gel electrophoresis systems, (b) elution from a particulate fraction in low ionic strength buffer, (c) co-migration with actin in isoelectric focusing, and (d) binding to DNase I. The identity of actin and its appearance on the platelet surface after thrombin-induced secretion was also demonstrated by the greater binding of an anti-actin antibody to thrombin-treated platelets, measured with 125I-staphylococcal protein A. Therefore, major platelet membrane changes occur after secretion but not after reversible aggregation. The platelet surface changes occurring with secretion may be important in the formation of irreversible platelet aggregates and in the final retraction of the blood clot. Images PMID:6772667

  8. Active membranes studied by X-ray scattering.

    PubMed

    Giahi, A; El Alaoui Faris, M; Bassereau, P; Salditt, T

    2007-08-01

    In view of recent theories of "active" membranes, we have studied multilamellar phospholipid membrane stacks with reconstituted transmembrane protein bacteriorhodopsin (BR) under different illumination conditions by X-ray scattering. The light-active protein is considered as an active constituent which drives the system out of equilibrium and is predicted to change the collective fluctuation properties of the membranes. Using X-ray reflectivity, X-ray non-specular (diffuse) scattering, and grazing incidence scattering, we find no detectable change in the scattering curves when changing the illumination condition. In particular the intermembrane spacing d remains constant, after eliminating hydration-related artifacts by design of a suitable sample environment. The absence of any observable non-equilibrium effects in the experimental window is discussed in view of the relevant parameters and recent theories. PMID:17712523

  9. Cholesterol modulates alkaline phosphatase activity of rat intestinal microvillus membranes.

    PubMed

    Brasitus, T A; Dahiya, R; Dudeja, P K; Bissonnette, B M

    1988-06-25

    Experiments were conducted, using a nonspecific lipid transfer protein, to vary the cholesterol/phospholipid molar ratio of rat proximal small intestinal microvillus membranes in order to assess the possible role of cholesterol in modulating enzymatic activities of this plasma membrane. Cholesterol/phospholipid molar ratios from 0.71 to 1.30 were produced from a normal value of 1.05 by incubation with the transfer protein and an excess of either phosphatidylcholine or cholesterol/phosphatidylcholine liposomes for 60 min at 37 degrees C. Cholesterol loading or depletion of the membranes was accompanied by a decrease or increase, respectively, in their lipid fluidity, as assessed by steady-state fluorescence polarization techniques using the lipid-soluble fluorophore 1,6-diphenyl-1,3,5-hexatriene. Increasing the cholesterol/phospholipid molar ratio also decreased alkaline phosphatase specific activity by approximately 20-30%, whereas decreasing this ratio increased this enzymatic activity by 20-30%. Sucrase, maltase, and lactase specific activities were not affected in these same preparations. Since the changes in alkaline phosphatase activity could be secondary to alterations in fluidity, cholesterol, or both, additional experiments were performed using benzyl alcohol, a known fluidizer. Benzyl alcohol (25 mM) restored the fluidity of cholesterol-enriched preparations to control levels, did not change the cholesterol/phospholipid molar ratio, and failed to alter alkaline phosphatase activity. These findings, therefore, indicate that alterations in the cholesterol content and cholesterol/phospholipid molar ratio of microvillus membranes can modulate alkaline phosphatase but not sucrase, maltase, or lactase activities. Moreover, membrane fluidity does not appear to be an important physiological regulator of these enzymatic activities. PMID:3379034

  10. Membrane Thinning and Thickening Induced by Membrane-Active Amphipathic Peptides.

    PubMed

    Grage, Stephan L; Afonin, Sergii; Kara, Sezgin; Buth, Gernot; Ulrich, Anne S

    2016-01-01

    Membrane thinning has been discussed as a fundamental mechanism by which antimicrobial peptides can perturb cellular membranes. To understand which factors play a role in this process, we compared several amphipathic peptides with different structures, sizes and functions in their influence on the lipid bilayer thickness. PGLa and magainin 2 from X. laevis were studied as typical representatives of antimicrobial cationic amphipathic α-helices. A 1:1 mixture of these peptides, which is known to possess synergistically enhanced activity, allowed us to evaluate whether and how this synergistic interaction correlates with changes in membrane thickness. Other systems investigated here include the α-helical stress-response peptide TisB from E. coli (which forms membrane-spanning dimers), as well as gramicidin S from A. migulanus (a natural antibiotic), and BP100 (designer-made antimicrobial and cell penetrating peptide). The latter two are very short, with a circular β-pleated and a compact α-helical structure, respectively. Solid-state (2)H-NMR and grazing incidence small angle X-ray scattering (GISAXS) on oriented phospholipid bilayers were used as complementary techniques to access the hydrophobic thickness as well as the bilayer-bilayer repeat distance including the water layer in between. This way, we found that magainin 2, gramicidin S, and BP100 induced membrane thinning, as expected for amphiphilic peptides residing in the polar/apolar interface of the bilayer. PGLa, on the other hand, decreased the hydrophobic thickness only at very high peptide:lipid ratios, and did not change the bilayer-bilayer repeat distance. TisB even caused an increase in the hydrophobic thickness and repeat distance. When reconstituted as a mixture, PGLa and magainin 2 showed a moderate thinning effect which was less than that of magainin 2 alone, hence their synergistically enhanced activity does not seem to correlate with a modulation of membrane thickness. Overall, the absence of

  11. Spatially distinct and metabolically active membrane domain in mycobacteria.

    PubMed

    Hayashi, Jennifer M; Luo, Chu-Yuan; Mayfield, Jacob A; Hsu, Tsungda; Fukuda, Takeshi; Walfield, Andrew L; Giffen, Samantha R; Leszyk, John D; Baer, Christina E; Bennion, Owen T; Madduri, Ashoka; Shaffer, Scott A; Aldridge, Bree B; Sassetti, Christopher M; Sandler, Steven J; Kinoshita, Taroh; Moody, D Branch; Morita, Yasu S

    2016-05-10

    Protected from host immune attack and antibiotic penetration by their unique cell envelope, mycobacterial pathogens cause devastating human diseases such as tuberculosis. Seamless coordination of cell growth with cell envelope elongation at the pole maintains this barrier. Unraveling this spatiotemporal regulation is a potential strategy for controlling mycobacterial infections. Our biochemical analysis previously revealed two functionally distinct membrane fractions in Mycobacterium smegmatis cell lysates: plasma membrane tightly associated with the cell wall (PM-CW) and a distinct fraction of pure membrane free of cell wall components (PMf). To provide further insight into the functions of these membrane fractions, we took the approach of comparative proteomics and identified more than 300 proteins specifically associated with the PMf, including essential enzymes involved in cell envelope synthesis such as a mannosyltransferase, Ppm1, and a galactosyltransferase, GlfT2. Furthermore, comparative lipidomics revealed the distinct lipid composition of the PMf, with specific association of key cell envelope biosynthetic precursors. Live-imaging fluorescence microscopy visualized the PMf as patches of membrane spatially distinct from the PM-CW and notably enriched in the pole of the growing cells. Taken together, our study provides the basis for assigning the PMf as a spatiotemporally distinct and metabolically active membrane domain involved in cell envelope biogenesis. PMID:27114527

  12. Experimental and numerical investigation of ADP square crystal with large aperture in the new Final Optics Assembly under the non-critical phase matching

    NASA Astrophysics Data System (ADS)

    Sun, Fuzhong; Zhang, Peng; Bai, Qingshun; Lu, Lihua; Xiang, Yong

    2016-04-01

    This paper presented a new Final Optics Assembly (FOA) of ammonium dihydrogen phosphate (ADP) square crystal with large aperture under the non-critical phase matching (NCPM), which controlled by the constant temperature water, and the temperature distribution was analyzed by simulation and experiment. Firstly, thermal analysis was carried out, as well as the temperature distribution of the cavity only heated under different velocities was analyzed. Then, the temperature distributions of ADP square crystal in the cavity were achieved using the Finite Volume Method (FVM), and this prediction was validated by the experiment results when the velocity is 0.1 m/s and 0.5 m/s. Finally, the optimal FHG conversion efficiency was obtained and the comparison of different heating methods was also highlighted.

  13. The effect of air flow on the temperature distribution and the harmonic conversion efficiency of the ADP crystal with large aperture in the temperature control scheme

    NASA Astrophysics Data System (ADS)

    Sun, Fuzhong; Zhang, Peng; Lu, Lihua; Xiang, Yong; Bai, Qingshun

    2016-03-01

    This paper presented a temperature control scheme for ammonium dihydrogen phosphate (ADP) crystal of Ф80 mm in diameter, and the influence of the air flow was also studied. This research aims to obtain the high energy, high frequency laser with large aperture under the non-critical phase matching (NCPM). Firstly, thermal analysis was carried out to investigate the air flow property in the cavity, as well as the effect of ambient temperature was analyzed. Secondly, the temperature distributions of air flow were achieved using the Finite Volume Method (FVM), and this prediction was validated by the experiment results. Finally, the effect of air flow in the cavity was obtained from the heating method, and the variation of harmonic conversion efficiency caused by the ambient temperature was also highlighted.

  14. Research on the Problem of High-Precision Deployment for Large-Aperture Space-Based Science Instruments

    NASA Technical Reports Server (NTRS)

    Lake, Mark S.; Peterson, Lee D.; Hachkowski, M. Roman; Hinkle, Jason D.; Hardaway, Lisa R.

    1998-01-01

    The present paper summarizes results from an ongoing research program conducted jointly by the University of Colorado and NASA Langley Research Center since 1994. This program has resulted in general guidelines for the design of high-precision deployment mechanisms, and tests of prototype deployable structures incorporating these mechanisms have shown microdynamically stable behavior (i.e., dimensional stability to parts per million). These advancements have resulted from the identification of numerous heretofore unknown microdynamic and micromechanical response phenomena, and the development of new test techniques and instrumentation systems to interrogate these phenomena. In addition, recent tests have begun to interrogate nanomechanical response of materials and joints and have been used to develop an understanding of nonlinear nanodynamic behavior in microdynamically stable structures. The ultimate goal of these efforts is to enable nano-precision active control of micro-precision deployable structures (i.e., active control to a resolution of parts per billion).

  15. Measuring Cysteine Cathepsin Activity to Detect Lysosomal Membrane Permeabilization.

    PubMed

    Repnik, Urška; Česen, Maruša Hafner; Turk, Boris

    2016-01-01

    During lysosomal membrane permeabilization (LMP), lysosomal lumenal contents can be released into the cytosol. Small molecules are more likely to be released, and cysteine cathepsins, with mature forms possessing a mass of 25-30 kDa, are among the smallest lumenal lysosomal enzymes. In addition, specific substrates for cysteine cathepsins are available to investigators, and therefore the measurement of the cathepsin activity as a hallmark of LMP works well. Here, we present a protocol for measuring the activity of these enzymes after selective plasma membrane permeabilization with a low concentration of digitonin and after total cell membrane lysis with a high concentration of digitonin. A fluorogenic substrate can be added either directly to the well with lysed cells to show LMP or to the cell-free extract to show that the lysosomal membrane has been sufficiently destabilized to allow the translocation of lysosomal enzymes. Although the content of lysosomal cysteine cathepsins differs between cell lines, this method has general applicability, is sensitive, and has high throughput. The presented protocol shows how to measure cysteine cathepsin activity in the presence of lysed cells and also in cell-free extracts. Depending on the aim of the study, one or both types of measurements can be performed. PMID:27140915

  16. Membrane phase characteristics control NA-CATH activity.

    PubMed

    Samuel, Robin; Gillmor, Susan

    2016-09-01

    Our studies presented in this report focus on the behavior of NA-CATH, an α-helical cathelicidin antimicrobial peptide, originally discovered in the Naja atra snake. It has demonstrated high potency against gram-positive and gram-negative bacteria with minimal hemolysis. Here we examine the kinetics, behaviors and potential mechanisms of the peptide in the presence of membrane liposome, modeling Escherichia coli, whose membrane exhibits distinct lipid phases. To understand NA-CATH interactions, the role of lipid phases is critical. We test three different lipid compositions to detangle the effect of phase on NA-CATH's activity using a series of leakage experiments. From these studies, we observe that NA-CATH changes from membrane disruption to pore-based lysing, depending on phases and lipid composition. This behavior also plays a major role in its kinetics. PMID:27216315

  17. Design and prototype tests of a large-aperture 37-53 MHz ferrite-tuned booster synchrotron cavity

    SciTech Connect

    Mark S. Champion et al.

    2001-07-12

    The Booster synchrotron at Fermilab employs eighteen 37-53 MHz ferrite-tuned double-gap coaxial radiofrequency cavities for acceleration of protons from 400 MeV to 8 GeV. The cavities have an aperture of 2.25 inches and operate at 55 kV per cavity. Future high duty factor operation of the Booster will be problematic due to unavoidable beam loss at the cavities resulting in excessive activation. The power amplifiers, high maintenance items, are mounted directly to the cavities in the tunnel. A proposed replacement for the Booster, the Proton Driver, will utilize the Booster radiofrequency cavities and requires not only a larger aperture, but also higher voltage. A research and development program is underway at Fermilab to modify the Booster cavities to provide a 5-inch aperture and a 20% voltage increase. A prototype has been constructed and high power tests have bee completed. The cavity design and test results is presented.

  18. Investigation of membrane active properties and antiradical activity of gossypol and its derivatives

    Technology Transfer Automated Retrieval System (TEKTRAN)

    New asymmetrical derivatives of gossypol were synthesized. The antioxidant activity of gossypol and these derivatives was studied. The interaction of these compounds with modeled lipid membranes was also studied. It was found that the antioxidant effects and ability to interact with membranes was...

  19. Membrane-Active Properties and Antiradical Activity of Gossypol and Its Derivatives

    Technology Transfer Automated Retrieval System (TEKTRAN)

    New asymmetrical derivatives of gossypol were synthesized. The antioxidant activity of gossypol and these derivatives was studied. The interaction of these compounds with modeled lipid membranes was also studied. It was found that the antioxidant effects and ability to interact with membranes was...

  20. HI-CLASS on AEOS: a large-aperture laser radar for space surveillance/situational awareness investigations

    NASA Astrophysics Data System (ADS)

    Kovacs, Mark A.; Dryden, Gordon L.; Pohle, Richard H.; Ayers, Kirstie; Carreras, Richard A.; Crawford, Linda L.; Taft, Russell

    2001-12-01

    The Air Force Research Laboratory/Directed Energy Directorate (AFRL/DE) via the ALVA (Applications of Lidars for Vehicles with Analysis) program installed in late 2000 a wideband, 12 J 15 Hz CO2 laser radar (ladar) on the 3.67 meter aperture AEOS (Advanced Electro-Optics System) telescope. This system is part of the Maui Space Surveillance System (MSSS), on the summit of Haleakala, Maui, HI. This ladar adopts the technology successfully demonstrated by the first generation HI-CLASS (High Performance CO2) Ladar Surveillance Sensor) operating on the nearby 0.6 meter aperture Laser Beam Director (LBD) and developed under the Field Ladar Demonstration program, jointly sponsored by AFRL/DE and the Army's Space and Missile Defense Command. The moderate power (approximately 180 watts) HI-CLASS/AEOS system generates multiple, coherent waveforms for precision satellite tracking and characterization of space objects for 1 m2 targets at ranges out to 10,000 km. This system also will be used to track space objects smaller than30 cm at ranges to 2,000 km. A third application of this system is to provide data for developing satellite identification, characterization, health and status techniques. This paper will discuss the operating characteristics and innovative features of the new system. The paper will also review recent results in support of AF needs, demonstrations, experiments, as well as planned activities that directly support applications in the DoD, scientific, and commercial arenas.

  1. Large Aperture Electrostatic Dust Detector

    SciTech Connect

    C.H. Skinner, R. Hensley, and A.L Roquemore

    2007-10-09

    Diagnosis and management of dust inventories generated in next-step magnetic fusion devices is necessary for their safe operation. A novel electrostatic dust detector, based on a fine grid of interlocking circuit traces biased to 30 or 50 ν has been developed for the detection of dust particles on remote surfaces in air and vacuum environments. Impinging dust particles create a temporary short circuit and the resulting current pulse is recorded by counting electronics. Up to 90% of the particles are ejected from the grid or vaporized suggesting the device may be useful for controlling dust inventories. We report measurements of the sensitivity of a large area (5x5 cm) detector to microgram quantities of dust particles and review its applications to contemporary tokamaks and ITER.

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

    PubMed

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

    2016-05-01

    Plant plasma membrane H(+)-ATPase acts as a primary transporter via proton pumping and regulates diverse physiological responses by controlling secondary solute transport, pH homeostasis, and membrane potential. Phosphorylation of the penultimate threonine and the subsequent binding of 14-3-3 proteins in the carboxyl terminus of the enzyme are required for H(+)-ATPase activation. We showed previously that photosynthesis induces phosphorylation of the penultimate threonine in the nonvascular bryophyte Marchantia polymorpha However, (1) whether this response is conserved in vascular plants and (2) the process by which photosynthesis regulates H(+)-ATPase phosphorylation at the plasma membrane remain unresolved issues. Here, we report that photosynthesis induced the phosphorylation and activation of H(+)-ATPase in Arabidopsis (Arabidopsis thaliana) leaves via sugar accumulation. Light reversibly phosphorylated leaf H(+)-ATPase, and this process was inhibited by pharmacological and genetic suppression of photosynthesis. Immunohistochemical and biochemical analyses indicated that light-induced phosphorylation of H(+)-ATPase occurred autonomously in mesophyll cells. We also show that the phosphorylation status of H(+)-ATPase and photosynthetic sugar accumulation in leaves were positively correlated and that sugar treatment promoted phosphorylation. Furthermore, light-induced phosphorylation of H(+)-ATPase was strongly suppressed in a double mutant defective in ADP-glucose pyrophosphorylase and triose phosphate/phosphate translocator (adg1-1 tpt-2); these mutations strongly inhibited endogenous sugar accumulation. Overall, we show that photosynthesis activated H(+)-ATPase via sugar production in the mesophyll cells of vascular plants. Our work provides new insight into signaling from chloroplasts to the plasma membrane ion transport mechanism. PMID:27016447

  3. Cyclohexane triones, novel membrane-active antibacterial agents.

    PubMed Central

    Lloyd, W J; Broadhurst, A V; Hall, M J; Andrews, K J; Barber, W E; Wong-Kai-In, P

    1988-01-01

    The cyclohexane triones are a novel group of synthetic antibacterial agents that are active against gram-positive bacteria, Haemophilus influenzae, and Mycobacterium smegmatis. In general, these compounds behaved in a manner similar to that of hexachlorophene, inhibiting the transport of low-molecular-weight hydrophilic substances into bacteria. Unlike cationic detergents, such as chlorhexidine, they did not cause disruption of the bacterial cytoplasmic membrane over a short time period. The most potent antibacterial cyclohexane trione studied had a reduced ability to inhibit solute transport in comparison with certain less active analogs. Cyclohexane triones may express more than a single type of antibacterial effect. PMID:3137860

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

    NASA Technical Reports Server (NTRS)

    Caldwell, C.

    1983-01-01

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

  5. Chemical Modulation of the Biological Activity of Reutericyclin: a Membrane-Active Antibiotic from Lactobacillus reuteri

    PubMed Central

    Cherian, Philip T.; Wu, Xiaoqian; Maddox, Marcus M.; Singh, Aman P.; Lee, Richard E.; Hurdle, Julian G.

    2014-01-01

    Whilst the development of membrane-active antibiotics is now an attractive therapeutic concept, progress in this area is disadvantaged by poor knowledge of the structure-activity relationship (SAR) required for optimizing molecules to selectively target bacteria. This prompted us to explore the SAR of the Lactobacillus reuteri membrane-active antibiotic reutericyclin, modifying three key positions about its tetramic acid core. The SAR revealed that lipophilic analogs were generally more active against Gram-positive pathogens, but introduction of polar and charged substituents diminished their activity. This was confirmed by cytometric assays showing that inactive compounds failed to dissipate the membrane potential. Radiolabeled substrate assays indicated that dissipation of the membrane potential by active reutericyclins correlated with inhibition of macromolecular synthesis in cells. However, compounds with good antibacterial activities also showed cytotoxicity against Vero cells and hemolytic activity. Although this study highlights the challenge of optimizing membrane-active antibiotics, it shows that by increasing antibacterial potency the selectivity index could be widened, allowing use of lower non-cytotoxic doses. PMID:24739957

  6. Large aperture Fizeau interferometer commissioning and preliminary measurements of a long x-ray mirror at European X-ray Free Electron Laser

    NASA Astrophysics Data System (ADS)

    Vannoni, M.; Freijo Martín, I.

    2016-05-01

    The European XFEL (X-ray Free Electron Laser) is a large facility under construction in Hamburg, Germany. It will provide a transversally fully coherent x-ray radiation with outstanding characteristics: high repetition rate (up to 2700 pulses with a 0.6 ms long pulse train at 10 Hz), short wavelength (down to 0.05 nm), short pulse (in the femtoseconds scale), and high average brilliance (1.6 ṡ 1025 (photons s-1 mm-2 mrad-2)/0.1% bandwidth). The beam has very high pulse energy; therefore, it has to be spread out on a relatively long mirror (about 1 m). Due to the very short wavelength, the mirrors need to have a high quality surface on their entire length, and this is considered very challenging even with the most advanced polishing methods. In order to measure the mirrors and to characterize their interaction with the mechanical mount, we equipped a metrology laboratory with a large aperture Fizeau interferometer. The system is a classical 100 mm diameter commercial Fizeau, with an additional expander providing a 300 mm diameter beam. Despite the commercial nature of the system, special care has been taken in the polishing of the reference flats and in the expander quality. We report the first commissioning of the instrument, its calibration, and performance characterization, together with some preliminary results with the measurement of a 950 mm silicon substrate. The intended application is to characterize the final XFEL mirrors with nanometer accuracy.

  7. Long-Term Evaluation of the Scintec Boundary-Layer Scintillometer and the Wageningen Large-Aperture Scintillometer: Implications for Scintillometer Users

    NASA Astrophysics Data System (ADS)

    Van Kesteren, B.; Beyrich, F.; Hartogensis, O. K.; Braam, M.

    2015-08-01

    We compare the structure parameter of the refractive index, , measured simultaneously with two large-aperture scintillometers: the WagLAS (Wageningen University, Wageningen, the Netherlands) and the BLS900 (Scintec, Rottenburg, Germany). A 3.5-year dataset shows a bias in of about 17 % between the instruments. Analysis of these data reveals firstly that the logarithmic amplifiers in the WagLAS exhibit a strong dependence on temperature, resulting in an overestimation of of up to 35 % for temperatures 0 . Secondly, high-pass filtering of the WagLAS and BLS900 intensity data artificially reduces for crosswinds 2 (error 25 and 5 % respectively). Thirdly, the BLS900 increasingly underestimates (up to 10-15 %) with increasing signal saturation. We demonstrate that Scintec's data processing relies too heavily on the assumption that the intensity data obey a log-normal distribution, which they do not in the case of saturation. Fourthly, both instruments ignore the dissipation range of the refractive-index spectrum, which leads to an overestimation of of up to 30 % for friction velocity 0.2 . Implications of these findings are discussed and placed into perspective for other scintillometer users. Furthermore, we present a tool for revealing saturation and other violations of Rytov theory for any given scintillometer type, including microwave scintillometers.

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

    NASA Astrophysics Data System (ADS)

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

    2012-02-01

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

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

    PubMed Central

    Singh, Prabhakar; Kesharwani, Rajesh Kumar; Misra, Krishna; Rizvi, Syed Ibrahim

    2016-01-01

    Plasma membrane redox system (PMRS) is an electron transport chain system ubiquitously present throughout all cell types. It transfers electron from intracellular substrates to extracellular acceptors for regulation of redox status. Curcumin, isolated from Curcuma longa, has modulatory effects on cellular physiology due to its membrane interaction ability and antioxidant potential. The present study investigates the effect of curcumin on PMRS activity of erythrocytes isolated from Wistar rats in vitro and in vivo and validated through an in silico docking simulation study using Molegro Virtual Docker (MVD). Effects of curcumin were also evaluated on level of glutathione (GSH) and the oxidant potential of plasma measured in terms of plasma ferric equivalent oxidative potentials (PFEOP). Results show that curcumin significantly (p < 0.01) downregulated the PMRS activity in a dose-dependent manner. Molecular docking results suggest that curcumin interacts with amino acids at the active site cavity of cytochrome b5 reductase, a key constituent of PMRS. Curcumin also increased the GSH level in erythrocytes and plasma while simultaneously decreasing the oxidant potential (PFEOP) of plasma. Altered PMRS activity and redox status are associated with the pathophysiology of several health complications including aging and diabetes; hence, the above finding may explain part of the role of curcumin in health beneficial effects. PMID:26904287

  10. Detecting Extracellular Carbonic Anhydrase Activity Using Membrane Inlet Mass Spectrometry

    PubMed Central

    Delacruz, Joannalyn; Mikulski, Rose; Tu, Chingkuang; Li, Ying; Wang, Hai; Shiverick, Kathleen T.; Frost, Susan C.; Horenstein, Nicole A.; Silverman, David N.

    2010-01-01

    Current research into the function of carbonic anhydrases in cell physiology emphasizes the role of membrane-bound carbonic anhydrases, such as carbonic anhydrase IX that has been identified in malignant tumors and is associated with extracellular acidification as a response to hypoxia. We present here a mass spectrometric method to determine the extent to which total carbonic anhydrase activity is due to extracellular carbonic anhydrase in whole cell preparations. The method is based on the biphasic rate of depletion of 18O from CO2 measured by membrane inlet mass spectrometry. The slopes of the biphasic depletion are a sensitive measure of the presence of carbonic anhydrase outside and inside of the cells. This property is demonstrated here using suspensions of human red cells in which external carbonic anhydrase was added to the suspending solution. It is also applied to breast and prostate cancer cells which both express exofacial carbonic anhydrase IX. Inhibition of external carbonic anhydrase is achieved by use of a membrane impermeant inhibitor that was synthesized for this purpose, p-aminomethylbenzenesulfonamide attached to a polyethyleneglycol polymer. PMID:20417171

  11. Membrane-Active Peptides and the Clustering of Anionic Lipids

    PubMed Central

    Wadhwani, P.; Epand, R.F.; Heidenreich, N.; Bürck, J.; Ulrich, A.S.; Epand, R.M.

    2012-01-01

    There is some overlap in the biological activities of cell-penetrating peptides (CPPs) and antimicrobial peptides (AMPs). We compared nine AMPs, seven CPPs, and a fusion peptide with regard to their ability to cluster anionic lipids in a mixture mimicking the cytoplasmic membrane of Gram-negative bacteria, as measured by differential scanning calorimetry. We also studied their bacteriostatic effect on several bacterial strains, and examined their conformational changes upon membrane binding using circular dichroism. A remarkable correlation was found between the net positive charge of the peptides and their capacity to induce anionic lipid clustering, which was independent of their secondary structure. Among the peptides studied, six AMPs and four CPPs were found to have strong anionic lipid clustering activity. These peptides also had bacteriostatic activity against several strains (particularly Gram-negative Escherichia coli) that are sensitive to lipid clustering agents. AMPs and CPPs that did not cluster anionic lipids were not toxic to E. coli. As shown previously for several types of AMPs, anionic lipid clustering likely contributes to the mechanism of antibacterial action of highly cationic CPPs. The same mechanism could explain the escape of CPPs from intracellular endosomes that are enriched with anionic lipids. PMID:22853904

  12. Hydrodynamic collective effects of active proteins in biological membranes.

    PubMed

    Koyano, Yuki; Kitahata, Hiroyuki; Mikhailov, Alexander S

    2016-08-01

    Lipid bilayers forming biological membranes are known to behave as viscous two-dimensional fluids on submicrometer scales; usually they contain a large number of active protein inclusions. Recently, it was shown [A. S. Mikhailov and R. Kapral, Proc. Natl. Acad. Sci. USA 112, E3639 (2015)PNASA60027-842410.1073/pnas.1506825112] that such active proteins should induce nonthermal fluctuating lipid flows leading to diffusion enhancement and chemotaxislike drift for passive inclusions in biomembranes. Here, a detailed analytical and numerical investigation of such effects is performed. The attention is focused on the situations when proteins are concentrated within lipid rafts. We demonstrate that passive particles tend to become attracted by active rafts and are accumulated inside them. PMID:27627343

  13. Effect of clofibrate on the enzyme activity of rat liver plasma membranes.

    PubMed

    Renaud, G; Foliot, A; Marais, J; Infante, R

    1980-03-15

    The activity of 3 plasma membranes marker enzymes (5'-nucleotidase, Mg++-ATPase and alkaline phosphodiesterase-I) was determined in plasma membranes isolated from liver of control and of clofibrate-treated rats. A complete indentity of plasma membranes enzyme activity in the 2 groups of experimental animals was observed for the 3 enzymes studied. PMID:6102923

  14. OpTIIX: An ISS-Based Testbed Paving the Roadmap Toward a Next Generation Large Aperture UV/Optical Space Telescope

    NASA Technical Reports Server (NTRS)

    Carpenter, Kenneth G.; Etemad, Shar; Seery, Bernard D.; Thronson, Harley; Burdick, Gary M.; Coulter, Dan; Goullioud, Renaud; Green, Joseph J.; Liu, Fengchuan; Ess, Kim; Postman, Marc; Sparks, Williams

    2012-01-01

    The next generation large aperture UV/Optical space telescope will need a diameter substantially larger than even that of JWST in order to address some of the most compelling unanswered scientific quests. These quests include understanding the earliest phases of the Universe and detecting life on exo-planets by studying spectra of their atmospheres. Such 8-16 meter telescopes face severe challenges in terms of cost and complexity and are unlikely to be affordable unless a new paradigm is adopted for their design and construction. The conventional approach is to use monolithic or preassembled segmented mirrors requiring complicated and risky deployments and relying on future heavy-lift vehicles, large fairings and complex geometry. The new paradigm is to launch component modules on relatively small vehicles and then perform in-orbit robotic assembly of those modules. The Optical Testbed and Integration on ISS eXperiment (OpTIIX) is designed to demonstrate, at low cost by leveraging the infrastructure provided by ISS, telescope assembly technologies and end-to-end optical system technologies. The use of ISS as a testbed permits the concentration of resources on reducing the technical risks associated with robotically integrating the components. These include laser metrology and wavefront sensing and control (WFS&C) systems, an imaging instrument, lightweight, low-cost deformable primary mirror segments and the secondary mirror. These elements are then aligned to a diffraction-limited optical system in space. The capability to assemble the optical system and remove and replace components via the existing ISS robotic systems like the Special Purpose Dexterous Manipulator (SPDM), or by the ISS flight crew, allows for future experimentation, as well as repair.

  15. Source locations of teleseismic P, SV, and SH waves observed in microseisms recorded by a large aperture seismic array in China

    NASA Astrophysics Data System (ADS)

    Liu, Qiaoxia; Koper, Keith D.; Burlacu, Relu; Ni, Sidao; Wang, Fuyun; Zou, Changqiao; Wei, Yunhao; Gal, Martin; Reading, Anya M.

    2016-09-01

    Transversely polarized seismic waves are routinely observed in ambient seismic energy across a wide range of periods, however their origin is poorly understood because the corresponding source regions are either undefined or weakly constrained, and nearly all models of microseism generation incorporate a vertically oriented single force as the excitation mechanism. To better understand the origin of transversely polarized energy in the ambient seismic wavefield we make the first systematic attempt to locate the source regions of teleseismic SH waves observed in microseismic (2.5-20 s) noise. We focus on body waves instead of surface waves because the source regions can be constrained in both azimuth and distance using conventional array techniques. To locate microseismic sources of SH waves (as well as SV and P waves) we continuously backproject the vertical, radial, and transverse components of the ambient seismic wavefield recorded by a large-aperture array deployed in China during 2013-2014. As expected, persistent P wave sources are observed in the North Atlantic, North Pacific, and Indian Oceans, mainly at periods of 2.5-10 s, in regions with the strong ocean wave interactions needed to produce secondary microseisms. SV waves are commonly observed to originate from locations indistinguishable from the P wave sources, but with smaller signal-to-noise ratios. We also observe SH waves with about half or less the signal-to-noise ratio of SV waves. SH source regions are definitively located in deep water portions of the Pacific, away from the sloping continental shelves that are thought to be important for the generation of microseismic Love waves, but nearby regions that routinely generate teleseismic P waves. The excitation mechanism for the observed SH waves may therefore be related to the interaction of P waves with small-wavelength bathymetric features, such as seamounts and basins, through some sort of scattering process.

  16. Lachesana tarabaevi, an expert in membrane-active toxins.

    PubMed

    Kuzmenkov, Alexey I; Sachkova, Maria Y; Kovalchuk, Sergey I; Grishin, Eugene V; Vassilevski, Alexander A

    2016-08-15

    In the present study, we show that venom of the ant spider Lachesana tarabaevi is unique in terms of molecular composition and toxicity. Whereas venom of most spiders studied is rich in disulfide-containing neurotoxic peptides, L. tarabaevi relies on the production of linear (no disulfide bridges) cytolytic polypeptides. We performed full-scale peptidomic examination of L. tarabaevi venom supported by cDNA library analysis. As a result, we identified several dozen components, and a majority (∼80% of total venom protein) exhibited membrane-active properties. In total, 33 membrane-interacting polypeptides (length of 18-79 amino acid residues) comprise five major groups: repetitive polypeptide elements (Rpe), latarcins (Ltc), met-lysines (MLys), cyto-insectotoxins (CIT) and latartoxins (LtTx). Rpe are short (18 residues) amphiphilic molecules that are encoded by the same genes as antimicrobial peptides Ltc 4a and 4b. Isolation of Rpe confirms the validity of the iPQM (inverted processing quadruplet motif) proposed to mark the cleavage sites in spider toxin precursors that are processed into several mature chains. MLys (51 residues) present 'idealized' amphiphilicity when modelled in a helical wheel projection with sharply demarcated sectors of hydrophobic, cationic and anionic residues. Four families of CIT (61-79 residues) are the primary weapon of the spider, accounting for its venom toxicity. Toxins from the CIT 1 and 2 families have a modular structure consisting of two shorter Ltc-like peptides. We demonstrate that in CIT 1a, these two parts act in synergy when they are covalently linked. This finding supports the assumption that CIT have evolved through the joining of two shorter membrane-active peptides into one larger molecule. PMID:27287558

  17. Basement membrane stiffening promotes retinal endothelial activation associated with diabetes.

    PubMed

    Yang, Xiao; Scott, Harry A; Monickaraj, Finny; Xu, Jun; Ardekani, Soroush; Nitta, Carolina F; Cabrera, Andrea; McGuire, Paul G; Mohideen, Umar; Das, Arup; Ghosh, Kaustabh

    2016-02-01

    Endothelial activation is a hallmark of the high-glucose (HG)-induced retinal inflammation associated with diabetic retinopathy (DR). However, precisely how HG induces retinal endothelial activation is not fully understood. We hypothesized that HG-induced up-regulation of lysyl oxidase (LOX), a collagen-cross-linking enzyme, in retinal capillary endothelial cells (ECs) enhances subendothelial basement membrane (BM) stiffness, which, in turn, promotes retinal EC activation. Diabetic C57BL/6 mice exhibiting a 70 and 50% increase in retinal intercellular adhesion molecule (ICAM)-1 expression and leukocyte accumulation, respectively, demonstrated a 2-fold increase in the levels of BM collagen IV and LOX, key determinants of capillary BM stiffness. Using atomic force microscopy, we confirmed that HG significantly enhances LOX-dependent subendothelial matrix stiffness in vitro, which correlated with an ∼2.5-fold increase in endothelial ICAM-1 expression, a 4-fold greater monocyte-EC adhesion, and an ∼2-fold alteration in endothelial NO (decrease) and NF-κB activation (increase). Inhibition of LOX-dependent subendothelial matrix stiffening alone suppressed HG-induced retinal EC activation. Finally, using synthetic matrices of tunable stiffness, we demonstrated that subendothelial matrix stiffening is necessary and sufficient to promote EC activation. These findings implicate BM stiffening as a critical determinant of HG-induced retinal EC activation and provide a rationale for examining BM stiffness and underlying mechanotransduction pathways as therapeutic targets for diabetic retinopathy. PMID:26443820

  18. Influence of decavanadate on rat synaptic plasma membrane ATPases activity.

    PubMed

    Krstić, Danijela; Colović, Mirjana; Bosnjaković-Pavlović, Nada; Spasojević-De Bire, Anne; Vasić, Vesna

    2009-09-01

    The in vitro influence of decameric vanadate species on Na+/K+-ATPase, plasma membrane Ca2+-ATPase (PMCA)-calcium pump and ecto-ATPase activity, using rat synaptic plasma membrane (SPM) as model system was investigated, whereas the commercial porcine cerebral cortex Na+/K+-ATPase served as a reference. The thermal behaviour of the synthesized decavanadate (V10) has been studied by differential scanning calorimetry and thermogravimetric analysis, while the type of polyvanadate anion was identified using the IR spectroscopy. The concentration-dependent responses to V10 of all enzymes were obtained. The half-maximum inhibitory concentration (IC50) of the enzyme activity was achieved at (4.74 +/- 1.15) x 10(-7) mol/l for SPM Na+/K+-ATPase, (1.30 +/- 0.10) x 10(-6) mol/l for commercial Na+/K+-ATPase and (3.13 +/- 1.70) x 10(-8) mol/l for Ca2+-ATPase, while ecto-ATPase is significantly less sensitive toward V10 (IC50 = (1.05 +/- 0.10) x 10(-4) mol/l) than investigated P-type ATPases. Kinetic analysis showed that V10 inhibited Na+/K+-ATPase by reducing the maximum enzymatic velocity and apparent affinity for ATP (increasing K(m) value), implying a mixed mode of interaction between V10 and P-type ATPases. PMID:20037196

  19. Conformational activation of visual rhodopsin in native disc membranes.

    PubMed

    Malmerberg, Erik; M Bovee-Geurts, Petra H; Katona, Gergely; Deupi, Xavier; Arnlund, David; Wickstrand, Cecilia; Johansson, Linda C; Westenhoff, Sebastian; Nazarenko, Elena; Schertler, Gebhard F X; Menzel, Andreas; de Grip, Willem J; Neutze, Richard

    2015-03-10

    Rhodopsin is the G protein-coupled receptor (GPCR) that serves as a dim-light receptor for vision in vertebrates. We probed light-induced conformational changes in rhodopsin in its native membrane environment at room temperature using time-resolved wide-angle x-ray scattering. We observed a rapid conformational transition that is consistent with an outward tilt of the cytoplasmic portion of transmembrane helix 6 concomitant with an inward movement of the cytoplasmic portion of transmembrane helix 5. These movements were considerably larger than those reported from the basis of crystal structures of activated rhodopsin, implying that light activation of rhodopsin involves a more extended conformational change than was previously suggested. PMID:25759477

  20. Membrane stretch and cytoplasmic Ca2+ independently modulate stretch-activated BK channel activity.

    PubMed

    Zhao, Hu-Cheng; Agula, Hasi; Zhang, Wei; Wang, Fa; Sokabe, Masahiro; Li, Lu-Ming

    2010-11-16

    Large conductance Ca(2+)-activated K(+) (BK) channels are responsible for changes in chemical and physical signals such as Ca(2+), Mg(2+) and membrane potentials. Previously, we reported that a BK channel cloned from chick heart (SAKCaC) is activated by membrane stretch. Molecular cloning and subsequent functional characterization of SAKCaC have shown that both the membrane stretch and intracellular Ca(2+) signal allosterically regulate the channel activity via the linker of the gating ring complex. Here we investigate how these two gating principles interact with each other. We found that stretch force activated SAKCaC in the absence of cytoplasmic Ca(2+). Lack of Ca(2+) bowl (a calcium binding motif) in SAKCaC diminished the Ca(2+)-dependent activation, but the mechanosensitivity of channel was intact. We also found that the abrogation of STREX (a proposed mechanosensing apparatus) in SAKCaC abolished the mechanosensitivity without altering the Ca(2+) sensitivity of channels. These observations indicate that membrane stretch and intracellular Ca(2+) could independently modulate SAKCaC activity. PMID:20673577

  1. Enzymatic activation of cellulose acetate membrane for reducing of protein fouling.

    PubMed

    Koseoglu-Imer, Derya Y; Dizge, Nadir; Koyuncu, Ismail

    2012-04-01

    In this study, the surface of cellulose acetate (CA) ultrafiltration membrane was activated with serine protease (Savinase) enzyme to reduce protein fouling. Enzyme molecules were covalently immobilized with glutaraldehyde (cross-linking agent) onto the surface of CA membranes. The membrane activation was verified using filtration experiments and morphological analysis. Scanning electron microscopy (SEM) images and attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy of the activated membrane when compared with raw membrane were confirmed that the enzyme was immobilized onto the membrane surface. The immobilization efficiencies changed from 13.2 to 41.2% according to the enzyme ratios from 2.5 to 10.0 mg/mL. However, the permeability values decreased from 232±6 to 121±4 L/m(2) h bar with increasing enzyme concentration from 2.5 to 10.0 mg/mL. In fouling experiments, bovine serum albumin (BSA) was used as the protein model solution and activated sludge was used as the model biological sludge. Enzyme-activated membranes exhibited good filtration performances and protein rejection efficiencies were compared with raw CA membrane. Also the relative flux reduction (RFR) ratios of membranes were calculated as 97% and 88% for raw CA and enzyme-activated membranes (5 mg/mL savinase), respectively. The membrane activated with Savinase enzyme could be proposed as a surface treatment method before filtration to mitigate protein fouling. PMID:22218336

  2. Cassava root membrane proteome reveals activities during storage root maturation.

    PubMed

    Naconsie, Maliwan; Lertpanyasampatha, Manassawe; Viboonjun, Unchera; Netrphan, Supatcharee; Kuwano, Masayoshi; Ogasawara, Naotake; Narangajavana, Jarunya

    2016-01-01

    Cassava (Manihot esculenta Crantz) is one of the most important crops of Thailand. Its storage roots are used as food, feed, starch production, and be the important source for biofuel and biodegradable plastic production. Despite the importance of cassava storage roots, little is known about the mechanisms involved in their formation. This present study has focused on comparison of the expression profiles of cassava root proteome at various developmental stages using two-dimensional gel electrophoresis and LC-MS/MS. Based on an anatomical study using Toluidine Blue, the secondary growth was confirmed to be essential during the development of cassava storage root. To investigate biochemical processes occurring during storage root maturation, soluble and membrane proteins were isolated from storage roots harvested from 3-, 6-, 9-, and 12-month-old cassava plants. The proteins with differential expression pattern were analysed and identified to be associated with 8 functional groups: protein folding and degradation, energy, metabolism, secondary metabolism, stress response, transport facilitation, cytoskeleton, and unclassified function. The expression profiling of membrane proteins revealed the proteins involved in protein folding and degradation, energy, and cell structure were highly expressed during early stages of development. Integration of these data along with the information available in genome and transcriptome databases is critical to expand knowledge obtained solely from the field of proteomics. Possible role of identified proteins were discussed in relation with the activities during storage root maturation in cassava. PMID:26547558

  3. An analysis on the influence of spatial scales on sensible heat fluxes in the north Tibetan Plateau based on Eddy covariance and large aperture scintillometer data

    NASA Astrophysics Data System (ADS)

    Sun, Genhou; Hu, Zeyong; Sun, Fanglin; Wang, Jiemin; Xie, Zhipeng; Lin, Yun; Huang, Fangfang

    2016-05-01

    The influence of spatial scales on surface fluxes is an interesting but not fully investigated question. This paper presents an analysis on the influence of spatial scales on surface fluxes in the north Tibetan Plateau based on eddy covariance (EC) and large aperture scintillometer (LAS) data at site Nagqu/BJ, combined with the land surface temperature (LST) and normalized difference vegetation index (NDVI) of moderate-resolution imaging spectroradiometer (MODIS). The analysis shows that sensible heat fluxes calculated with LAS data (H_LAS) agree reasonably well with sensible heat fluxes calculated with EC data (H_EC) in the rain and dry seasons. The difference in their footprints due to the wind direction is an important reason for the differences in H_EC and H_LAS. The H_LAS are statistically more consistent with H_EC when their footprints overlap than when their footprints do not. A detailed analysis on H_EC and H_LAS changes with net radiation and wind direction in rain and dry season indicates that the spatial heterogeneity in net radiation created by clouds contributes greatly to the differences in H_EC and H_LAS in short-term variations. A significant relationship between the difference in footprint-weighted averages of LST and difference in H_EC and H_LAS suggests that the spatial heterogeneity in LST at two spatial scales is a reason for the differences in H_EC and H_LAS and that LST has a positive correlation with the differences in H_EC and H_LAS. A significant relationship between the footprint-weighted averages of NDVI and the ratio of sensible heat fluxes at two spatial scales to net radiation (H/Rn) in the rain season supports the analysis that the spatial heterogeneity in canopy at two spatial scales is another reason for differences in H_EC and H_LAS and that canopy has a negative correlation with (H/Rn). An analysis on the influence of the difference in aerodynamic roughness lengths at two spatial scales on sensible heat fluxes shows that the

  4. A comparison of detection sensitivity between ALTAIR and Arecibo meteor observations: Can high power and large aperture radars detect low velocity meteor head-echoes

    NASA Astrophysics Data System (ADS)

    Janches, Diego; Close, Sigrid; Fentzke, Jonathan T.

    2008-01-01

    Meteor head-echo observations using High Power and Large Aperture (HPLA) radars have been routinely used for micrometeor studies for over a decade. The head-echo is a signal from the radar-reflective plasma region traveling with the meteoroid and its detection allows for very precise determination of instantaneous meteor altitude, velocity and deceleration. Unlike specular meteor radars (SMR), HPLA radars are diverse instruments when compared one to another. The operating frequencies range from 46 MHz to 1.29 GHz while the antenna configurations changes from 18,000 dipoles in a 300 m×300 m square array, phase arrays of dipoles to single spherical or parabolic dishes of various dimensions. Hunt et al. [Hunt, S.M., Oppenheim, M., Close, S., Brown, P.G., McKeen, F., Minardi, M., 2004. Icarus 168, 34-42] and Close et al. [Close, S., Brown, P., Campbell-Brown, M., Oppenheim, M., Colestock, P., 2007. Icarus, doi: 10.1016/j.icarus.2006.09.07] recently showed, by utilizing a head-echo plasma-based model, the presence of instrumental biases in the ALTAIR VHF radar system against detecting meteors produced by very small particles (<1 μg) moving at slow (˜20 km/s) velocities due to the low head echo radar cross-section (RCS) associated with these particles. In this paper we apply the same methodology to the Arecibo 430 MHz radar and compare the results with those presented by Close et al. [Close, S., Brown, P., Campbell-Brown, M., Oppenheim, M., Colestock, P., 2007. Icarus, doi: 10.1016/j.icarus.2006.09.07]. We show that, if the methodology applied by Hunt et al. [Hunt, S.M., Oppenheim, M., Close, S., Brown, P.G., McKeen, F., Minardi, M., 2004. Icarus 168, 34-42] and Close et al. [Close, S., Brown, P., Campbell-Brown, M., Oppenheim, M., Colestock, P., 2007. Icarus, doi: 10.1016/j.icarus.2006.09.07] is accurate, for particles at least 1 μg or heavier, while the bias may exist for the ALTAIR measurements, it does not exist in the Arecibo data due to its greater sensitivity.

  5. Functional, photochemically active, and chemically asymmetric membranes by interfacial polymerization of derivatized multifunctional prepolymers

    DOEpatents

    Lonsdale, H.K.; Wamser, C.C.

    1990-04-17

    The preparation of a novel class of thin film membranes by interfacial polymerization is disclosed, said membranes incorporating as part of their polymeric structure the functionality of monomeric or oligomeric precursors. Specific embodiments include porphyrin and phthalocyanine derivatives that are photochemically or electrochemically active, as well as chemically asymmetric membranes.

  6. Functional, photochemically active, and chemically asymmetric membranes by interfacial polymerization of derivatized multifunctional prepolymers

    DOEpatents

    Lonsdale, Harold K.; Wamser, Carl C.

    1990-01-01

    The preparation of a novel class of thin film membranes by interfacial polymerization is disclosed, said membranes incorporating as part of their polymeric structure the functionality of monomeric or oligomeric precursors. Specific embodiments include porphyrin and phthalocyanine derivatives that are photochemically or electrochemically active, as well as chemically asymmetric membranes.

  7. Fusicoccin Binding to Its Plasma Membrane Receptor and the Activation of the Plasma Membrane H+-ATPase

    PubMed Central

    De Michelis, Maria Ida; Pugliarello, Maria Chiara; Rasi-Caldogno, Franca

    1989-01-01

    The characteristics of fusicoccin binding were investigated in microsomes from 24-h-old radish (Raphanus sativus L.) seedlings. The time course of fusicoccin binding depended on fusicoccin concentration: equilibrium was reached much faster at 10 nanomolar fusicoccin than at 0.3 nanomolar fusicoccin. Scatchard analysis of equilibrium binding as a function of fusicoccin concentration indicated a single class of receptor sites with a Kd of 1.8 nanomolar and a site density of 6.3 picomoles per milligram protein. Similar values (Kd 1.7 nanomolar and site density 7 picomoles per milligram protein) were obtained from the analysis of the dependence of equilibrium binding on membrane concentration at fixed fusicoccin concentrations. Fusicoccin binding comigrated with the plasma membrane H+-ATPase in an equilibrium sucrose density gradient: both activities formed a sharp peak (1.18 grams per milliliter) clearly distinct from that of markers of other membranes which all peaked at lower densities. The saturation profiles of fusicoccin binding and of fusicoccin-induced activation of the plasma membrane H+-ATPase, measured under identical conditions, were similar, supporting the view that fusicoccin-induced activation of the plasma membrane H+-ATPase is mediated by fusicoccin binding to its plasma membrane receptor. PMID:16666723

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

    PubMed

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

    2016-05-01

    In the present study, we report that polyphenols present in red wine obtained by a controlled microvinification process are able to protect human erythrocytes from oxidative stress and to activate Plasma Membrane Redox System (PMRS). Human plasma obtained from healthy subjects was incubated in the presence of whole red wine at a concentration corresponding to 9.13-73 μg/ml gallic acid equivalents to verify the capacity to protect against hypochlorous acid (HOCl)-induced plasma oxidation and to minimize chloramine formation. Red wine reduced hemolysis and chloramine formation induced by HOCl of 40 and 35%, respectively. PMRS present on human erythrocytes transfers electrons from intracellular molecules to extracellular electron acceptors. We demonstrated that whole red wine activated PMRS activity in human erythrocytes isolated from donors in a dose-dependent manner with a maximum at about 70-100 μg/ml gallic acid equivalents. We also showed that red wine increased glutathione (GSH) levels and erythrocytic antioxidant capacity, measured by 2,2-diphenyl-1-picrylhydrazyl (DPPH) quenching assay. Furthermore, we reported that GSH played a crucial role in regulating PMRS activity in erythrocytes. In fact, the effect of iodoacetamide, an alkylating agent that induces depletion of intracellular GSH, was completely counteracted by red wine. Bioactive compounds present in red wine, such as gallic acid, resveratrol, catechin, and quercetin were unable to activate PMRS when tested at the concentrations normally present in aged red wines. On the contrary, the increase of PMRS activity was associated with the anthocyanin fraction, suggesting the capacity of this class of compounds to positively modulate PMRS enzymatic activity. PMID:26866566

  9. The Lipopeptide Antibiotic Paenibacterin Binds to the Bacterial Outer Membrane and Exerts Bactericidal Activity through Cytoplasmic Membrane Damage

    PubMed Central

    Huang, En

    2014-01-01

    Paenibacterin is a broad-spectrum lipopeptide antimicrobial agent produced by Paenibacillus thiaminolyticus OSY-SE. The compound consists of a cyclic 13-residue peptide and an N-terminal C15 fatty acyl chain. The mechanism of action of paenibacterin against Escherichia coli and Staphylococcus aureus was investigated in this study. The cationic lipopeptide paenibacterin showed a strong affinity for the negatively charged lipopolysaccharides (LPS) from the outer membrane of Gram-negative bacteria. Addition of LPS (100 μg/ml) completely eliminated the antimicrobial activity of paenibacterin against E. coli. The electrostatic interaction between paenibacterin and LPS may have displaced the divalent cations on the LPS network and thus facilitated the uptake of antibiotic into Gram-negative cells. Paenibacterin also damaged the bacterial cytoplasmic membrane, as evidenced by the depolarization of membrane potential and leakage of intracellular potassium ions from cells of E. coli and S. aureus. Therefore, the bactericidal activity of paenibacterin is attributed to disruption of the outer membrane of Gram-negative bacteria and damage of the cytoplasmic membrane of both Gram-negative and Gram-positive bacteria. Despite the evidence of membrane damage, this study does not rule out additional bactericidal mechanisms potentially exerted by paenibacterin. PMID:24561581

  10. NPA binding activity is peripheral to the plasma membrane and is associated with the cytoskeleton.

    PubMed Central

    Cox, D N; Muday, G K

    1994-01-01

    N-1-Naphthylphthalamic acid (NPA) binding activity is released into the supernatant when plasma membranes are subjected to high-salt treatment, indicating that this activity is peripherally associated with the membrane. Extraction of plasma membrane vesicles with Triton X-100 resulted in retention of NPA binding activity in the detergent-insoluble cytoskeletal pellet. Treatment of this pellet with KI released NPA binding activity, actin, and alpha-tubulin. Dialysis to remove KI led to the repolymerization of cytoskeletal elements and movement of NPA binding activity into an insoluble cytoskeletal pellet. NPA binding activity partitioned into the detergent-insoluble cytoskeletal pellet obtained from both zucchini and maize membranes and was released from these pellets by KI treatment. Treatment of a cytoskeletal pellet with cytochalasin B doubled NPA binding activity in the resulting supernatant. Together, these experiments indicate that NPA binding activity is peripherally associated with the plasma membrane and interacts with the cytoskeleton in vitro. PMID:11536654

  11. Voltage-induced membrane displacement in patch pipettes activates mechanosensitive channels

    PubMed Central

    Gil, Ziv; Silberberg, Shai D.; Magleby, Karl L.

    1999-01-01

    The patch-clamp technique allows currents to be recorded through single ion channels in patches of cell membrane in the tips of glass pipettes. When recording, voltage is typically applied across the membrane patch to drive ions through open channels and to probe the voltage-sensitivity of channel activity. In this study, we used video microscopy and single-channel recording to show that prolonged depolarization of a membrane patch in borosilicate pipettes results in delayed slow displacement of the membrane into the pipette and that this displacement is associated with the activation of mechanosensitive (MS) channels in the same patch. The membrane displacement, ≈1 μm with each prolonged depolarization, occurs after variable delays ranging from tens of milliseconds to many seconds and is correlated in time with activation of MS channels. Increasing the voltage step shortens both the delay to membrane displacement and the delay to activation. Preventing depolarization-induced membrane displacement by applying positive pressure to the shank of the pipette or by coating the tips of the borosilicate pipettes with soft glass prevents the depolarization-induced activation of MS channels. The correlation between depolarization-induced membrane displacement and activation of MS channels indicates that the membrane displacement is associated with sufficient membrane tension to activate MS channels. Because membrane tension can modulate the activity of various ligand and voltage-activated ion channels as well as some transporters, an apparent voltage dependence of a channel or transporter in a membrane patch in a borosilicate pipette may result from voltage-induced tension rather than from direct modulation by voltage. PMID:10588750

  12. Enzymatically active high-flux selectively gas-permeable membranes

    DOEpatents

    Jiang, Ying-Bing; Cecchi, Joseph L.; Rempe, Susan; FU, Yaqin; Brinker, C. Jeffrey

    2016-01-26

    An ultra-thin, catalyzed liquid transport medium-based membrane structure fabricated with a porous supporting substrate may be used for separating an object species such as a carbon dioxide object species. Carbon dioxide flux through this membrane structures may be several orders of magnitude higher than traditional polymer membranes with a high selectivity to carbon dioxide. Other gases such as molecular oxygen, molecular hydrogen, and other species including non-gaseous species, for example ionic materials, may be separated using variations to the membrane discussed.

  13. How Lipid Membranes Affect Pore Forming Toxin Activity.

    PubMed

    Rojko, Nejc; Anderluh, Gregor

    2015-12-15

    Pore forming toxins (PFTs) evolved to permeate the plasma membrane of target cells. This is achieved in a multistep mechanism that usually involves binding of soluble protein monomer to the lipid membrane, oligomerization at the plane of the membrane, and insertion of part of the polypeptide chain across the lipid membrane to form a conductive channel. Introduced pores allow uncontrolled transport of solutes across the membrane, inflicting damage to the target cell. PFTs are usually studied from the perspective of structure-function relationships, often neglecting the important role of the bulk membrane properties on the PFT mechanism of action. In this Account, we discuss how membrane lateral heterogeneity, thickness, and fluidity influence the pore forming process of PFTs. In general, lipid molecules are more accessible for binding in fluid membranes due to steric reasons. When PFT specifically binds ordered domains, it usually recognizes a specific lipid distribution pattern, like sphingomyelin (SM) clusters or SM/cholesterol complexes, and not individual lipid species. Lipid domains were also suggested to act as an additional concentration platform facilitating PFT oligomerization, but this is yet to be shown. The last stage in PFT action is the insertion of the transmembrane segment across the membranes to build the transmembrane pore walls. Conformational changes are a spontaneous process, and sufficient free energy has to be available for efficient membrane penetration. Therefore, fluid bilayers are permeabilized more readily in comparison to highly ordered and thicker liquid ordered lipid phase (Lo). Energetically more costly insertion into the Lo phase can be driven by the hydrophobic mismatch between the thinner liquid disordered phase (Ld) and large protein complexes, which are unable to tilt like single transmembrane segments. In the case of proteolipid pores, membrane properties can directly modulate pore size, stability, and even selectivity. Finally

  14. Activation of a heat-stable cytolytic protein associated with the surface membrane of Naegleria fowleri.

    PubMed Central

    Lowrey, D M; McLaughlin, J

    1985-01-01

    Surface membrane-enriched fractions of Naegleria fowleri obtained after isopycnic centrifugation experiments contain a potent cytolytic activity as determined by hemolysis and 51Cr release assays. This surface membrane cytolysin was unaffected by a treatment at 75 degrees C for 30 min and accounted for 70 to 90% of cytolysis by whole-cell lysates of amoebae. This heat resistance as well as intimate membrane association distinguished the surface membrane cytolytic activity from a second heat-labile cytolytic activity which appears to be latent within lysosomes. The surface membrane cytolysin was found to be specifically activated by diluted samples of lysosomal fractions. The possible role of this surface membrane cytotoxin in the pathogenicity of N. fowleri is discussed. PMID:4055029

  15. Alterations in the activities of hepatic plasma-membrane and microsomal enzymes during liver regeneration.

    PubMed Central

    Deliconstantinos, G; Ramantanis, G

    1983-01-01

    A marked increase in the activities of rat liver plasma-membrane (Na+ + K+)-stimulated ATPase and microsomal Ca2+-stimulated ATPase was observed 18h after partial hepatectomy. Lipid analyses for both membrane preparations reveal that in partially hepatectomized rats the cholesterol and sphingomyelin content are decreased with a subsequent decrease in the cholesterol/phospholipid molar ratio compared with those of sham-operated animals. Changes in the allosteric properties of plasma-membrane (Na+ + K+)-stimulated ATPase by F- (as reflected by changes in the Hill coefficient) indicated a fluidization of the lipid bilayer of both membrane preparations in 18 h-regenerating liver. The amphipathic dodecyl glucoside incorporated into the hepatic plasma membranes evoked a marked increase in the (Na+ + K+)-stimulated ATPase and 5'-nucleotidase activities. The lack of effect of the glucoside on the Lubrol-PX-solubilized 5'-nucleotidase indicates that changes in the activities of the membrane-bound enzymes caused by the glucoside are due to modulation of the membrane fluidity. Dodecyl glucoside appears to increase the membrane fluidity, evaluated through changes in the Hill coefficient for plasma-membrane (Na+ + K+)-stimulated ATPase. The biological significance of these data is discussed in terms of the differences and changes in the interaction of membrane-bound enzymes with membrane lipids during liver regeneration. PMID:6309144

  16. ULTRASTRUCTURE OF VEILLONELLA AND MORPHOLOGICAL CORRELATION OF AN OUTER MEMBRANE WITH PARTICLES ASSOCIATED WITH ENDOTOXIC ACTIVITY

    PubMed Central

    Bladen, Howard A.; Mergenhagen, Stephan E.

    1964-01-01

    Bladen, Howard A. (National Institute of Dental Research, Bethesda, Md.), and Stephan E. Mergenhagen. Ultrastructure of Veillonella and morphological correlation of an outer membrane with particles associated with endotoxic activity. J. Bacteriol. 88:1482–1492. 1964.—Normal, phenol-water extracted, and lysozyme-treated Veillonella cells were embedded in Vestopal W, sectioned, and examined by electron microscopy. Normal cells as well as the phenol-water extract (endotoxin) were examined by negative and positive contrast techniques. In thin sections of normal cells, three separate structural entities were observed surrounding the protoplasm, and were referred to as the outer membrane, the solid membrane, and the plasma membrane. The outer membrane was a membrane composed of two dense layers (30 A) separated by a less-dense layer (20 A), and followed a convoluted and continuous path around the cell. The solid membrane appeared as a taut, dense structure 100 to 500 A wide, and was separated from the outer membrane by up to several hundred Ångstroms. The plasma membrane was a unit-type membrane. After cells were treated with phenol-water, the outer membrane was absent, but the cells remained intact owing to the solid membrane. Observation of the phenol-water extract (endotoxin) revealed predominantly circular particles or discs which had approximately the same dimensions in height as the outer membrane had in width. Negatively stained whole cells showed similar structures on their surface. Lysozyme treatment of the cells did not affect the outer membrane; however, the solid membrane became diffuse and often disappeared, suggesting that the outer membrane and the solid membrane were separate structures. Images PMID:14234809

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

    SciTech Connect

    Cunningham, K.; Wickner, W.T. )

    1989-11-01

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

  18. Cholinesterase activity per unit surface area of conducting membranes.

    PubMed

    Brzin, M; Dettbarn, W D; Rosenberg, P; Nachmansohn, D

    1965-08-01

    According to theory, the action of acetylcholine (ACh) and ACh-esterase is essential for the permeability changes of excitable membranes during activity. It is, therefore, pertinent to know the activity of ACh-esterase per unit axonal surface area instead of per gram nerve, as it has been measured in the past. Such information has now been obtained with the newly developed microgasometric technique using a magnetic diver. (1) The cholinesterase (Ch-esterase) activity per mm(2) surface of sensory axons of the walking leg of lobster is 1.2 x 10(-3) microM/hr. (sigma = +/- 0.3 x 10(-3); SE = 0.17 x 10(-3)); the corresponding value for the motor axons isslightly higher: 1.93 x 10(-3) microM/hr. (sigma = +/- 0.41 x 10(-3); SE = +/- 0.14 x 10(-3)). Referred to gram nerve, the Ch-esterase activity of the sensory axons is much higher than that of the motor axons: 741 microM/hr. (sigma = +/- 73.5; SE = +/- 32.6) versus 111.6 microM/hr. (sigma = +/- 28.3; SE = +/- 10). (2) The enzyme activity in the small fibers of the stellar nerve of squid is 3.2 x 10(-4) microM/mm(2)/hr. (sigma = +/- 0.96 x 10(-4); SE = +/- 0.4 x 10(-4)). (3) The Ch-esterase activity per mm(2) surface of squid giant axon is 9.5 x 10(-5) microM/hr. (sigma = +/- 1.55 x 10(-5); SE = +/- 0.38 x 10(-5)). The value was obtained with small pieces of carefully cleaned axons after removal of the axoplasm and exposure to sonic disintegration. Without the latter treatment the figurewas 3.85 x 10(-5) microM/mm(2)/hr. (sigma = +/- 3.24 x 10(-5); SE = +/- 0.93 x 10(-5)). The experiments indicate the existence of permeability barriers in the cell wall surrounding part of the enzyme, since the substrate cannot reach all the enzyme even when small fragments of the cell wall are used without disintegration. (4) On the basis of the data obtained, some tentative approximations are made of the ratio of ACh released to Na ions entering the squid giant axon per cm(2) per impulse. PMID:5865929

  19. Pneumolysin Activates Macrophage Lysosomal Membrane Permeabilization and Executes Apoptosis by Distinct Mechanisms without Membrane Pore Formation

    PubMed Central

    Bewley, Martin A.; Naughton, Michael; Preston, Julie; Mitchell, Andrea; Holmes, Ashleigh; Marriott, Helen M.; Read, Robert C.; Mitchell, Timothy J.; Whyte, Moira K. B.

    2014-01-01

    ABSTRACT Intracellular killing of Streptococcus pneumoniae is complemented by induction of macrophage apoptosis. Here, we show that the toxin pneumolysin (PLY) contributes both to lysosomal/phagolysosomal membrane permeabilization (LMP), an upstream event programing susceptibility to apoptosis, and to apoptosis execution via a mitochondrial pathway, through distinct mechanisms. PLY is necessary but not sufficient for the maximal induction of LMP and apoptosis. PLY’s ability to induce both LMP and apoptosis is independent of its ability to form cytolytic pores and requires only the first three domains of PLY. LMP involves TLR (Toll-like receptor) but not NLRP3/ASC (nucleotide-binding oligomerization domain [Nod]-like receptor family, pyrin domain-containing protein 3/apoptosis-associated speck-like protein containing a caspase recruitment domain) signaling and is part of a PLY-dependent but phagocytosis-independent host response that includes the production of cytokines, including interleukin-1 beta (IL-1β). LMP involves progressive and selective permeability to 40-kDa but not to 250-kDa fluorescein isothiocyanate (FITC)-labeled dextran, as PLY accumulates in the cytoplasm. In contrast, the PLY-dependent execution of apoptosis requires phagocytosis and is part of a host response to intracellular bacteria that also includes NO generation. In cells challenged with PLY-deficient bacteria, reconstitution of LMP using the lysomotrophic detergent LeuLeuOMe favored cell necrosis whereas PLY reconstituted apoptosis. The results suggest that PLY contributes to macrophage activation and cytokine production but also engages LMP. Following bacterial phagocytosis, PLY triggers apoptosis and prevents macrophage necrosis as a component of a broad-based antimicrobial strategy. This illustrates how a key virulence factor can become the focus of a multilayered and coordinated innate response by macrophages, optimizing pathogen clearance and limiting inflammation. PMID:25293758

  20. Salt stress in a membrane bioreactor: dynamics of sludge properties, membrane fouling and remediation through powdered activated carbon dosing.

    PubMed

    De Temmerman, L; Maere, T; Temmink, H; Zwijnenburg, A; Nopens, I

    2014-10-15

    Membrane bioreactors are a well-established technology for wastewater treatment. However, their efficiency is adversely impacted by membrane fouling, primarily inciting very conservative operations of installations that makes them less appealing from an economic perspective. This fouling propensity of the activated sludge is closely related to system disturbances. Therefore, improved insight into the impact of fouling is crucial towards increased membrane performance. In this work, the disturbance of a salt shock was investigated with respect to sludge composition and filterability in two parallel lab-scale membrane bioreactors. Several key sludge parameters (soluble microbial products, sludge-bound extracellular polymeric substances, supramicron particle size distributions (PSD), submicron particle concentrations) were intensively monitored prior to, during, and after a disturbance to investigate its impact as well as the potential governing mechanism. Upon salt addition, the supramicron PSD immediately shifted to smaller floc sizes, and the total fouling rate increased. Following a certain delay, an increase in submicron particles, supernatant proteins, and polysaccharides was observed as well as an increase in the irreversible membrane fouling rate. Recovery from the disturbance was evidenced with a simultaneous decrease in the above mentioned quantities. A similar experiment introducing powdered activated carbon (PAC) addition used for remediation resulted in either no or less significant changes in the above mentioned quantities, signifying its potential as a mitigation strategy. PMID:24999116

  1. Circulating polymerase chain reaction chips utilizing multiple-membrane activation

    NASA Astrophysics Data System (ADS)

    Wang, Chih-Hao; Chen, Yi-Yu; Liao, Chia-Sheng; Hsieh, Tsung-Min; Luo, Ching-Hsing; Wu, Jiunn-Jong; Lee, Huei-Huang; Lee, Gwo-Bin

    2007-02-01

    This paper reports a new micromachined, circulating, polymerase chain reaction (PCR) chip for nucleic acid amplification. The PCR chip is comprised of a microthermal control module and a polydimethylsiloxane (PDMS)-based microfluidic control module. The microthermal control modules are formed with three individual heating and temperature-sensing sections, each modulating a specific set temperature for denaturation, annealing and extension processes, respectively. Micro-pneumatic valves and multiple-membrane activations are used to form the microfluidic control module to transport sample fluids through three reaction regions. Compared with other PCR chips, the new chip is more compact in size, requires less time for heating and cooling processes, and has the capability to randomly adjust time ratios and cycle numbers depending on the PCR process. Experimental results showed that detection genes for two pathogens, Streptococcus pyogenes (S. pyogenes, 777 bps) and Streptococcus pneumoniae (S. pneumoniae, 273 bps), can be successfully amplified using the new circulating PCR chip. The minimum number of thermal cycles to amplify the DNA-based S. pyogenes for slab gel electrophoresis is 20 cycles with an initial concentration of 42.5 pg µl-1. Experimental data also revealed that a high reproducibility up to 98% could be achieved if the initial template concentration of the S. pyogenes was higher than 4 pg µl-1. The preliminary results of the current paper were presented at the 19th IEEE International Conference on Micro Electro Mechanical Systems (IEEE MEMS 2006), Istanbul, Turkey, 22-26 January, 2006.

  2. Thylakoid Membrane Maturation and PSII Activation Are Linked in Greening Synechocystis sp. PCC 6803 Cells1

    PubMed Central

    Barthel, Sandra; Bernát, Gábor; Seidel, Tobias; Rupprecht, Eva; Kahmann, Uwe; Schneider, Dirk

    2013-01-01

    Thylakoid membranes are typical and essential features of both chloroplasts and cyanobacteria. While they are crucial for phototrophic growth of cyanobacterial cells, biogenesis of thylakoid membranes is not well understood yet. Dark-grown Synechocystis sp. PCC 6803 cells contain only rudimentary thylakoid membranes but still a relatively high amount of phycobilisomes, inactive photosystem II and active photosystem I centers. After shifting dark-grown Synechocystis sp. PCC 6803 cells into the light, “greening” of Synechocystis sp. PCC 6803 cells, i.e. thylakoid membrane formation and recovery of photosynthetic electron transport reactions, was monitored. Complete restoration of a typical thylakoid membrane system was observed within 24 hours after an initial lag phase of 6 to 8 hours. Furthermore, activation of photosystem II complexes and restoration of a functional photosynthetic electron transport chain appears to be linked to the biogenesis of organized thylakoid membrane pairs. PMID:23922268

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

    PubMed

    Purdy, P H; Barbosa, E A; Praamsma, C J; Schisler, G J

    2016-08-01

    We investigated the effects of two trout sperm activation solutions on sperm physiology and membrane organization prior to and following cryopreservation using flow cytometry and investigated their impact on in vitro fertility. Overall, frozen-thawed samples had greater phospholipid disorder when compared with fresh samples (high plasma membrane fluidity; P < 0.0001) and sperm activated with water also had high plasma membrane fluidity when compared to sperm activated with Lahnsteiner solution (LAS; P < 0.0001). Following cryopreservation water activated samples had membranes with greater membrane protein disorganization compared with LAS but the membrane protein organization of LAS samples was similar to samples prior to freezing (P < 0.0001). Post-thaw water activation resulted in significant increases in intracellular calcium compared to LAS (P < 0.002). In vitro fertility trials with frozen-thawed milt and LAS activation resulted in greater fertility (45%) compared to water activated samples (10%; P < 0.0001). Higher fertility rates correlated with lower intracellular calcium with water (R(2) = -0.9; P = 0.01) and LAS (R(2) = -0.85; P = 0.03) activation. Greater plasma membrane phospholipid (R(2) = -0.89; P = 0.02) and protein (R(2) = -0.84; P = 0.04) disorder correlated with lower water activation fertility rates. These membrane organization characteristics only approached significance with LAS activation in vitro fertility (P = 0.09, P = 0.06, respectively). Potentially the understanding of sperm membrane reorganizations and the physiology associated with activation following cryopreservation may enable users in a repository or hatchery setting to estimate the fertilizing potential of a sample and determine its value. PMID:27234987

  4. Actomyosin dynamics drive local membrane component organization in an in vitro active composite layer.

    PubMed

    Köster, Darius Vasco; Husain, Kabir; Iljazi, Elda; Bhat, Abrar; Bieling, Peter; Mullins, R Dyche; Rao, Madan; Mayor, Satyajit

    2016-03-22

    The surface of a living cell provides a platform for receptor signaling, protein sorting, transport, and endocytosis, whose regulation requires the local control of membrane organization. Previous work has revealed a role for dynamic actomyosin in membrane protein and lipid organization, suggesting that the cell surface behaves as an active composite composed of a fluid bilayer and a thin film of active actomyosin. We reconstitute an analogous system in vitro that consists of a fluid lipid bilayer coupled via membrane-associated actin-binding proteins to dynamic actin filaments and myosin motors. Upon complete consumption of ATP, this system settles into distinct phases of actin organization, namely bundled filaments, linked apolar asters, and a lattice of polar asters. These depend on actin concentration, filament length, and actin/myosin ratio. During formation of the polar aster phase, advection of the self-organizing actomyosin network drives transient clustering of actin-associated membrane components. Regeneration of ATP supports a constitutively remodeling actomyosin state, which in turn drives active fluctuations of coupled membrane components, resembling those observed at the cell surface. In a multicomponent membrane bilayer, this remodeling actomyosin layer contributes to changes in the extent and dynamics of phase-segregating domains. These results show how local membrane composition can be driven by active processes arising from actomyosin, highlighting the fundamental basis of the active composite model of the cell surface, and indicate its relevance to the study of membrane organization. PMID:26929326

  5. Comparison of microbial communities of activated sludge and membrane biofilm in 10 full-scale membrane bioreactors.

    PubMed

    Jo, Sung Jun; Kwon, Hyeokpil; Jeong, So-Yeon; Lee, Chung-Hak; Kim, Tae Gwan

    2016-09-15

    Operation of membrane bioreactors (MBRs) for wastewater treatment is hampered by the membrane biofouling resulting from microbial activities. However, the knowledge of the microbial ecology of both biofilm and activated sludge in MBRs has not been sufficient. In this study, we scrutinized microbial communities of biofilm and activated sludge from 10 full-scale MBR plants. Overall, Flavobacterium, Dechloromonas and Nitrospira were abundant in order of abundance in biofilm, whereas Dechloromonas, Flavobacterium and Haliscomenobacter in activated sludge. Community structure was analyzed in either biofilm or activated sludge. Among MBRs, as expected, not only diversity of microbial community but also its composition was different from one another (p < 0.05). Between the biofilm and activated sludge, community composition made significant difference, but its diversity measures (i.e., alpha diversity, e.g., richness, diversity and evenness) did not (p > 0.05). Effects of ten environmental factors on community change were investigated using Spearman correlation. MLSS, HRT, F/M ratio and SADm explained the variation of microbial composition in the biofilm, whereas only MLSS did in the activated sludge. Microbial networks were constructed with the 10 environmental factors. The network results revealed that there were different topological characteristics between the biofilm and activated sludge networks, in which each of the 4 factors had different associations with microbial nodes. These results indicated that the different microbial associations were responsible for the variation of community composition between the biofilm and activated sludge. PMID:27262549

  6. Spatial distribution and activity of Na(+)/K(+)-ATPase in lipid bilayer membranes with phase boundaries.

    PubMed

    Bhatia, Tripta; Cornelius, Flemming; Brewer, Jonathan; Bagatolli, Luis A; Simonsen, Adam C; Ipsen, John H; Mouritsen, Ole G

    2016-06-01

    We have reconstituted functional Na(+)/K(+)-ATPase (NKA) into giant unilamellar vesicles (GUVs) of well-defined binary and ternary lipid composition including cholesterol. The activity of the membrane system can be turned on and off by ATP. The hydrolytic activity of NKA is found to depend on membrane phase, and the water relaxation in the membrane on the presence of NKA. By collapsing and fixating the GUVs onto a solid support and using high-resolution atomic-force microscopy (AFM) imaging we determine the protein orientation and spatial distribution at the single-molecule level and find that NKA is preferentially located at lo/ld interfaces in two-phase GUVs and homogeneously distributed in single-phase GUVs. When turned active, the membrane is found to unbind from the support suggesting that the protein function leads to softening of the membrane. PMID:26994932

  7. New insights into membrane-active action in plasma membrane of fungal hyphae by the lipopeptide antibiotic bacillomycin L.

    PubMed

    Zhang, Bao; Dong, Chunjuan; Shang, Qingmao; Han, Yuzhu; Li, Pinglan

    2013-09-01

    Bacillomycin L, a natural iturinic lipopeptide produced by Bacillus amyloliquefaciens, is characterized by strong antifungal activities against a variety of agronomically important filamentous fungi including Rhizoctonia solani Kühn. Prior to this study, the role of membrane permeabilization in the antimicrobial activity of bacillomycin L against plant pathogenic fungi had not been investigated. To shed light on the mechanism of this antifungal activity, the permeabilization of R. solani hyphae by bacillomycin L was investigated and compared with that by amphotericin B, a polyene antibiotic which is thought to act primarily through membrane disruption. Our results derived from electron microscopy, various fluorescent techniques and gel retardation experiments revealed that the antifungal activity of bacillomycin L may be not solely a consequence of fungal membrane permeabilization, but related to the interaction of it with intracellular targets. Our findings provide more insights into the mode of action of bacillomycin L and other iturins, which could in turn help to develop new or improved antifungal formulations or result in novel strategies to prevent fungal spoilage. PMID:23756779

  8. Influence of activating hormones on human platelet membrane Ca/sup 2 +/-ATPase activity

    SciTech Connect

    Resink, T.J.; Dimitrov, D.; Stucki, S.; Buehler, F.R.

    1986-07-16

    Intact platelets were pretreated with hormones and thereafter membranes were prepared and Ca/sup 2 +/-ATPase activity determined. Thrombin decreased the V/sub max/ of Ca/sup 2 +/-ATPase after pretreatment of intact platelets. Platelet activating factor, vasopressin and ADP also decreased Ca/sup 2 +/-ATPase activity. 12-O-tetradecanoylphorbol-13-acetate (TPA) or A23187 or ionomycin alone had no effect, while the simultaneous pretreatment with TPA and Ca/sup 2 +/-ionophore decreased Ca/sup 2 +/-ATPase activity. cAMP elevating agents prostaglandin E/sub 1/ (PGE/sub 1/) and forskolin had no influence per se on Ca/sup 2 +/-ATPase, but antagonized the inhibitory effect of thrombin. The data suggest a close connection between phosphoinositide metabolism and the Ca/sup 2 +/-ATPase system.

  9. Functional, photochemically active, and chemically asymmetric membranes by interfacial polymerization of derivatized multifunctional prepolymers

    DOEpatents

    Lonsdale, Harold K.; Wamser, Carl C.

    1988-01-01

    The preparation of a novel class of thin film membranes by interfacial polymerization is disclosed, said membanes incorporating as part of their polymeric structure the functionality of monomeric or oligomeric precursors. Specific embodiments include porphyrin and phthalocyanime derivatives that are photochemically or electrochemically active, as well as chemically asymmetric membranes.

  10. One-way membrane trafficking of SOS in receptor-triggered Ras activation.

    PubMed

    Christensen, Sune M; Tu, Hsiung-Lin; Jun, Jesse E; Alvarez, Steven; Triplet, Meredith G; Iwig, Jeffrey S; Yadav, Kamlesh K; Bar-Sagi, Dafna; Roose, Jeroen P; Groves, Jay T

    2016-09-01

    SOS is a key activator of the small GTPase Ras. In cells, SOS-Ras signaling is thought to be initiated predominantly by membrane recruitment of SOS via the adaptor Grb2 and balanced by rapidly reversible Grb2-SOS binding kinetics. However, SOS has multiple protein and lipid interactions that provide linkage to the membrane. In reconstituted-membrane experiments, these Grb2-independent interactions were sufficient to retain human SOS on the membrane for many minutes, during which a single SOS molecule could processively activate thousands of Ras molecules. These observations raised questions concerning how receptors maintain control of SOS in cells and how membrane-recruited SOS is ultimately released. We addressed these questions in quantitative assays of reconstituted SOS-deficient chicken B-cell signaling systems combined with single-molecule measurements in supported membranes. These studies revealed an essentially one-way trafficking process in which membrane-recruited SOS remains trapped on the membrane and continuously activates Ras until being actively removed via endocytosis. PMID:27501536

  11. Active membrane transport and receptor proteins from bacteria.

    PubMed

    Saidijam, M; Bettaney, K E; Szakonyi, G; Psakis, G; Shibayama, K; Suzuki, S; Clough, J L; Blessie, V; Abu-Bakr, A; Baumberg, S; Meuller, J; Hoyle, C K; Palmer, S L; Butaye, P; Walravens, K; Patching, S G; O'reilly, J; Rutherford, N G; Bill, R M; Roper, D I; Phillips-Jones, M K; Henderson, P J F

    2005-08-01

    A general strategy for the expression of bacterial membrane transport and receptor genes in Escherichia coli is described. Expression is amplified so that the encoded proteins comprise 5-35% of E. coli inner membrane protein. Depending upon their topology, proteins are produced with RGSH6 or a Strep tag at the C-terminus. These enable purification in mg quantities for crystallization and NMR studies. Examples of one nutrient uptake and one multidrug extrusion protein from Helicobacter pylori are described. This strategy is successful for membrane proteins from H. pylori, E. coli, Enterococcus faecalis, Bacillus subtilis, Staphylococcus aureus, Microbacterium liquefaciens, Brucella abortus, Brucella melitensis, Campylobacter jejuni, Neisseria meningitides, Streptomyces coelicolor and Rhodobacter sphaeroides. PMID:16042616

  12. Lipid composition and sensitivity of Prototheca wickerhamii to membrane-active antimicrobial agents.

    PubMed Central

    Sud, I J; Feingold, D S

    1979-01-01

    The lipid composition of Prototheca wickerhamii ATCC 16529 is presented and discussed in relation to the unique susceptibility of the organism to drugs of three membrane-active antimicrobial classes: the polyenes, the polymyxins, and the imidazoles. The presence of ergosterol in the neutral lipid fraction of the membrane is likely responsible for the exquisite susceptibility to amphotericin B. The presence of a large quantity of free fatty acids in the membrane appears responsible for imidazole susceptibility. The membrane determinants of polymyxin B susceptibility are less well defined. PMID:518077

  13. Sar1 GTPase Activity Is Regulated by Membrane Curvature*♦

    PubMed Central

    Hanna, Michael G.; Mela, Ioanna; Wang, Lei; Henderson, Robert M.; Chapman, Edwin R.; Edwardson, J. Michael; Audhya, Anjon

    2016-01-01

    The majority of biosynthetic secretory proteins initiate their journey through the endomembrane system from specific subdomains of the endoplasmic reticulum. At these locations, coated transport carriers are generated, with the Sar1 GTPase playing a critical role in membrane bending, recruitment of coat components, and nascent vesicle formation. How these events are appropriately coordinated remains poorly understood. Here, we demonstrate that Sar1 acts as the curvature-sensing component of the COPII coat complex and highlight the ability of Sar1 to bind more avidly to membranes of high curvature. Additionally, using an atomic force microscopy-based approach, we further show that the intrinsic GTPase activity of Sar1 is necessary for remodeling lipid bilayers. Consistent with this idea, Sar1-mediated membrane remodeling is dramatically accelerated in the presence of its guanine nucleotide-activating protein (GAP), Sec23-Sec24, and blocked upon addition of guanosine-5′-[(β,γ)-imido]triphosphate, a poorly hydrolysable analog of GTP. Our results also indicate that Sar1 GTPase activity is stimulated by membranes that exhibit elevated curvature, potentially enabling Sar1 membrane scission activity to be spatially restricted to highly bent membranes that are characteristic of a bud neck. Taken together, our data support a stepwise model in which the amino-terminal amphipathic helix of GTP-bound Sar1 stably penetrates the endoplasmic reticulum membrane, promoting local membrane deformation. As membrane bending increases, Sar1 membrane binding is elevated, ultimately culminating in GTP hydrolysis, which may destabilize the bilayer sufficiently to facilitate membrane fission. PMID:26546679

  14. Active endocannabinoids are secreted on extracellular membrane vesicles.

    PubMed

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

    2015-02-01

    Endocannabinoids primarily influence neuronal synaptic communication within the nervous system. To exert their function, endocannabinoids need to travel across the intercellular space. However, how hydrophobic endocannabinoids cross cell membranes and move extracellularly remains an unresolved problem. Here, we show that endocannabinoids are secreted through extracellular membrane vesicles produced by microglial cells. We demonstrate that microglial extracellular vesicles carry on their surface N-arachidonoylethanolamine (AEA), which is able to stimulate type-1 cannabinoid receptors (CB1), and inhibit presynaptic transmission, in target GABAergic neurons. This is the first demonstration of a functional role of extracellular vesicular transport of endocannabinoids. PMID:25568329

  15. Molecular Details of Membrane Fluidity Changes during Apoptosis and Relationship to Phospholipase A2 Activity

    PubMed Central

    Gibbons, Elizabeth; Pickett, Katalyn R.; Streeter, Michael C.; Warcup, Ashley O.; Nelson, Jennifer; Judd, Allan M.; Bell, John D.

    2012-01-01

    Summary Secretory phospholipase A2 exhibits much greater activity toward apoptotic versus healthy cells. Various plasma membrane changes responsible for this phenomenon have been proposed, including biophysical alterations described as “membrane fluidity” and “order.” Understanding of these membrane perturbations was refined by applying studies with model membranes to fluorescence measurements during thapsigargin-induced apoptosis of S49 cells using probes specific for the plasma membrane: Patman and trimethylammonium-diphenylhexatriene. Alterations in emission properties of these probes corresponded with enhanced susceptibility of the cells to hydrolysis by secretory phospholipase A2. By applying a quantitative model, additional information was extracted from the kinetics of Patman equilibration with the membrane. Taken together, these data suggested that the phospholipids of apoptotic membranes display greater spacing between adjacent headgroups, reduced interactions between neighboring lipid tails, and increased penetration of water among the heads. The phase transition of artificial bilayers was used to calibrate quantitatively the relationship between probe fluorescence and the energy of interlipid interactions. This analysis was applied to results from apoptotic cells to estimate the frequency with which phospholipids protrude sufficiently at the membrane surface to enter the enzyme’s active site. The data suggested that this frequency increases 50–100-fold as membranes become susceptible to hydrolysis during apoptosis. PMID:22967861

  16. Uncoupling the Structure-Activity Relationships of β2 Adrenergic Receptor Ligands from Membrane Binding.

    PubMed

    Dickson, Callum J; Hornak, Viktor; Velez-Vega, Camilo; McKay, Daniel J J; Reilly, John; Sandham, David A; Shaw, Duncan; Fairhurst, Robin A; Charlton, Steven J; Sykes, David A; Pearlstein, Robert A; Duca, Jose S

    2016-06-23

    Ligand binding to membrane proteins may be significantly influenced by the interaction of ligands with the membrane. In particular, the microscopic ligand concentration within the membrane surface solvation layer may exceed that in bulk solvent, resulting in overestimation of the intrinsic protein-ligand binding contribution to the apparent/measured affinity. Using published binding data for a set of small molecules with the β2 adrenergic receptor, we demonstrate that deconvolution of membrane and protein binding contributions allows for improved structure-activity relationship analysis and structure-based drug design. Molecular dynamics simulations of ligand bound membrane protein complexes were used to validate binding poses, allowing analysis of key interactions and binding site solvation to develop structure-activity relationships of β2 ligand binding. The resulting relationships are consistent with intrinsic binding affinity (corrected for membrane interaction). The successful structure-based design of ligands targeting membrane proteins may require an assessment of membrane affinity to uncouple protein binding from membrane interactions. PMID:27239696

  17. Neomycin inhibits the phosphatidylinositol monophosphate and phosphatidylinositol bisphosphate stimulation of plasma membrane ATPase activity

    SciTech Connect

    Chen, Qiuyun; Boss, W.F. )

    1991-05-01

    The inositol phospholipids, phosphatidylinositol monophosphate (PIP) and phosphatidylinositol bisphosphate (PIP{sub 2}), have been shown to increase the vanadate-sensitive ATPase activity of plant plasma membranes. In this paper, the authors show the effect of various concentrations of phosphatidyinositol, PIP, and PIP{sub 2} on the plasma membrane vanadate-sensitive ATPase activity. PIP and PIP{sub 2} at concentrations at 10 nanomoles per 30 microgram membrane protein per milliliter of reaction mixture caused a twofold and 1.8-fold increase in the ATPase activity, respectively. The effect of these negatively charged phospholipids on the ATPase activity was inhibited by adding the positively charged aminoglycoside, neomycin. Neomycin did not affect the endogenous plasma membrane ATPase activity in the absence of exogenous lipids.

  18. Membrane lipids regulate ganglioside GM2 catabolism and GM2 activator protein activity.

    PubMed

    Anheuser, Susi; Breiden, Bernadette; Schwarzmann, Günter; Sandhoff, Konrad

    2015-09-01

    Ganglioside GM2 is the major lysosomal storage compound of Tay-Sachs disease. It also accumulates in Niemann-Pick disease types A and B with primary storage of SM and with cholesterol in type C. Reconstitution of GM2 catabolism with β-hexosaminidase A and GM2 activator protein (GM2AP) at uncharged liposomal surfaces carrying GM2 as substrate generated only a physiologically irrelevant catabolic rate, even at pH 4.2. However, incorporation of anionic phospholipids into the GM2 carrying liposomes stimulated GM2 hydrolysis more than 10-fold, while the incorporation of plasma membrane stabilizing lipids (SM and cholesterol) generated a strong inhibition of GM2 hydrolysis, even in the presence of anionic phospholipids. Mobilization of membrane lipids by GM2AP was also inhibited in the presence of cholesterol or SM, as revealed by surface plasmon resonance studies. These lipids also reduced the interliposomal transfer rate of 2-NBD-GM1 by GM2AP, as observed in assays using Förster resonance energy transfer. Our data raise major concerns about the usage of recombinant His-tagged GM2AP compared with untagged protein. The former binds more strongly to anionic GM2-carrying liposomal surfaces, increases GM2 hydrolysis, and accelerates intermembrane transfer of 2-NBD-GM1, but does not mobilize membrane lipids. PMID:26175473

  19. Role of liquid membrane phenomenon in the anti-bacterial activity of Cefuroxime Sodium

    PubMed Central

    Nagesh, C.; Shankaraiah, M. M.; Venkatesh, J. S.; Setty, S. Ramachandra

    2010-01-01

    The role of liquid membrane phenomenon has been studied in the anti bacterial activity of cephalosporins i.e. Cefuroxime sodium. In our earlier publication [1] it was reported that hydraulic permeability data obtained to demonstrate the existence of liquid membrane in series with supporting membrane generated by Cefuroxime sodium. Transport of selected permeants (glucose, PABA, glycine, and ions like Mg++, NH4+, PO4-, Ca++, Na+, K+ and Cl-) through liquid membrane generated by Cefuroxime sodium in series with supporting membrane has been studied. The results indicated that the liquid membrane generated by Cefuroxime sodium inhibit the transport of various essential bio-molecules and permeants in to the cell. This modification in permeability of different permeants in the presence of the liquid membranes is likely to play significant role in the biological actions of Cefuroxime sodium. The anti-bacterial activity of Cefuroxime sodium further confirmed that the generation of liquid membrane by Cefuroxime sodium is also contributing for the antibacterial activity of them. PMID:24825969

  20. Correlation of Daptomycin Bactericidal Activity and Membrane Depolarization in Staphylococcus aureus

    PubMed Central

    Silverman, Jared A.; Perlmutter, Nancy G.; Shapiro, Howard M.

    2003-01-01

    The objective of this study was to further elucidate the role of membrane potential in the mechanism of action of daptomycin, a novel lipopeptide antibiotic. Membrane depolarization was measured by both fluorimetric and flow cytometric assays. Adding daptomycin (5 μg/ml) to Staphylococcus aureus gradually dissipated membrane potential. In both assays, cell viability was reduced by >99% and membrane potential was reduced by >90% within 30 min of adding daptomycin. Cell viability decreased in parallel with changes in membrane potential, demonstrating a temporal correlation between bactericidal activity and membrane depolarization. Decreases in viability and potential also showed a dose-dependent correlation. Depolarization is indicative of ion movement across the cytoplasmic membrane. Fluorescent probes were used to demonstrate Ca2+-dependent, daptomycin-triggered potassium release from S. aureus. Potassium release was also correlated with bactericidal activity. This study demonstrates a clear correlation between dissipation of membrane potential and the bactericidal activity of daptomycin. A multistep model for daptomycin's mechanism of action is proposed. PMID:12878516

  1. Plasma Membrane ATPase Activity following Reversible and Irreversible Freezing Injury 1

    PubMed Central

    Iswari, S.; Palta, Jiwan P.

    1989-01-01

    Plasma membrane ATPase has been proposed as a site of functional alteration during early stages of freezing injury. To test this, plasma membrane was purified from Solanum leaflets by a single step partitioning of microsomes in a dextran-polyethylene glycol two phase system. Addition of lysolecithin in the ATPase assay produced up to 10-fold increase in ATPase activity. ATPase activity was specific for ATP with a Km around 0.4 millimolar. Presence of the ATPase enzyme was identified by immunoblotting with oat ATPase antibodies. Using the phase partitioning method, plasma membrane was isolated from Solanum commersonii leaflets which had four different degrees of freezing damage, namely, slight (reversible), partial (partially reversible), substantial and total (irreversible). With slight (reversible) damage the plasma membrane ATPase specific activity increased 1.5- to 2-fold and its Km was decreased by about 3-fold, whereas the specific activity of cytochrome c reductase and cytochrome c oxidase in the microsomes were not different from the control. However, with substantial (lethal, irreversible) damage, there was a loss of membrane protein, decrease in plasma membrane ATPase specific activity and decrease in Km, while cytochrome c oxidase and cytochrome c reductase were unaffected. These results support the hypothesis that plasma membrane ATPase is altered by slight freeze-thaw stress. Images Figure 1 Figure 2 PMID:16666856

  2. Tetraspanins regulate the protrusive activities of cell membrane

    SciTech Connect

    Bari, Rafijul; Guo, Qiusha; Zhongnan Hospital, Wuhan University, Wuhan ; Xia, Bing; Zhang, Yanhui H.; Giesert, Eldon E.; Levy, Shoshana; Zheng, Jie J.; Zhang, Xin A.

    2011-12-02

    Highlights: Black-Right-Pointing-Pointer Tetraspanins regulate microvillus formation. Black-Right-Pointing-Pointer Tetraspanin CD81 promotes microvillus formation. Black-Right-Pointing-Pointer Tetraspanin CD82 inhibits microvillus formation. Black-Right-Pointing-Pointer Based on this study, we extrapolated a general cellular mechanism for tetraspanins. Black-Right-Pointing-Pointer Tetraspanins engage various functions by regulating membrane protrusion morphogenesis. -- Abstract: Tetraspanins have gained increased attention due to their functional versatility. But the universal cellular mechanism that governs such versatility remains unknown. Herein we present the evidence that tetraspanins CD81 and CD82 regulate the formation and/or development of cell membrane protrusions. We analyzed the ultrastructure of the cells in which a tetraspanin is either overexpressed or ablated using transmission electron microscopy. The numbers of microvilli on the cell surface were counted, and the radii of microvillar tips and the lengths of microvilli were measured. We found that tetraspanin CD81 promotes the microvillus formation and/or extension while tetraspanin CD82 inhibits these events. In addition, CD81 enhances the outward bending of the plasma membrane while CD82 inhibits it. We also found that CD81 and CD82 proteins are localized at microvilli using immunofluorescence. CD82 regulates microvillus morphogenesis likely by altering the plasma membrane curvature and/or the cortical actin cytoskeletal organization. We predict that membrane protrusions embody a common morphological phenotype and cellular mechanism for, at least some if not all, tetraspanins. The differential effects of tetraspanins on microvilli likely lead to the functional diversification of tetraspanins and appear to correlate with their functional propensity.

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

    PubMed Central

    Wilmes, Miriam; Stockem, Marina; Bierbaum, Gabriele; Schlag, Martin; Götz, Friedrich; Tran, Dat Q.; Schaal, Justin B.; Ouellette, André J.; Selsted, Michael E.; Sahl, Hans-Georg

    2014-01-01

    θ-Defensins are cyclic antimicrobial peptides expressed in leukocytes of Old world monkeys. To get insight into their antibacterial mode of action, we studied the activity of RTDs (rhesus macaque θ-defensins) against staphylococci. We found that in contrast to other defensins, RTDs do not interfere with peptidoglycan biosynthesis, but rather induce bacterial lysis in staphylococci by interaction with the bacterial membrane and/or release of cell wall lytic enzymes. Potassium efflux experiments and membrane potential measurements revealed that the membrane impairment by RTDs strongly depends on the energization of the membrane. In addition, RTD treatment caused the release of Atl-derived cell wall lytic enzymes probably by interaction with membrane-bound lipoteichoic acid. Thus, the premature and uncontrolled activity of these enzymes contributes strongly to the overall killing by θ-defensins. Interestingly, a similar mode of action has been described for Pep5, an antimicrobial peptide of bacterial origin. PMID:25632351

  4. Immobilization and activity assay of cytochrome P450 on patterned lipid membranes

    SciTech Connect

    Ueda, Yoshihiro; Morigaki, Kenichi . E-mail: morigaki-kenichi@aist.go.jp; Tatsu, Yoshiro; Yumoto, Noboru; Imaishi, Hiromasa . E-mail: himaish@kobe-u.ac.jp

    2007-04-20

    We report on a methodology for immobilizing cytochrome P450 on the surface of micropatterned lipid bilayer membranes and measuring the enzymatic activity. The patterned bilayer comprised a matrix of polymeric lipid bilayers and embedded fluid lipid bilayers. The polymeric lipid bilayer domains act as a barrier to confine fluid lipid bilayers in defined areas and as a framework to stabilize embedded membranes. The fluid bilayer domains, on the other hand, can contain lipid compositions that facilitate the fusion between lipid membranes, and are intended to be used as the binding agent of microsomes containing rat CYP1A1. By optimizing the membrane compositions of the fluid bilayers, we could selectively immobilize microsomal membranes on these domains. The enzymatic activity was significantly higher on lipid bilayer substrates compared with direct adsorption on glass. Furthermore, competitive assay experiment between two fluorogenic substrates demonstrated the feasibility of bioassays based on immobilized P450s.

  5. Immobilization and activity assay of cytochrome P450 on patterned lipid membranes.

    PubMed

    Ueda, Yoshihiro; Morigaki, Kenichi; Tatsu, Yoshiro; Yumoto, Noboru; Imaishi, Hiromasa

    2007-04-20

    We report on a methodology for immobilizing cytochrome P450 on the surface of micropatterned lipid bilayer membranes and measuring the enzymatic activity. The patterned bilayer comprised a matrix of polymeric lipid bilayers and embedded fluid lipid bilayers. The polymeric lipid bilayer domains act as a barrier to confine fluid lipid bilayers in defined areas and as a framework to stabilize embedded membranes. The fluid bilayer domains, on the other hand, can contain lipid compositions that facilitate the fusion between lipid membranes, and are intended to be used as the binding agent of microsomes containing rat CYP1A1. By optimizing the membrane compositions of the fluid bilayers, we could selectively immobilize microsomal membranes on these domains. The enzymatic activity was significantly higher on lipid bilayer substrates compared with direct adsorption on glass. Furthermore, competitive assay experiment between two fluorogenic substrates demonstrated the feasibility of bioassays based on immobilized P450s. PMID:17335776

  6. Activated pathways for the directed insertion of patterned nanoparticles into polymer membranes.

    PubMed

    Ting, Christina L; Frischknecht, Amalie L

    2013-10-28

    We combine the string method with self-consistent field theory to compute the most probable transition pathway, i.e. the minimum free energy path, for the insertion of Janus and protein-like nanoparticles into a polymer membrane bilayer. The method makes no assumptions in the reaction coordinate and overcomes the long timescales challenge associated with simulating rare events. Our study suggests that one approach to building functional polymer–nanoparticle composite membranes with oriented nanoparticles is through electrostatic interactions. In particular, hydrophobic Janus nanoparticles with an asymmetric charge distribution can be made to directionally insert into charged membranes. This process is kinetically driven, and involves overcoming a thermally surmountable activation barrier, which requires favorable interactions between the nanoparticle and the hydrophilic block of the membrane. In contrast, the insertion of protein-like nanoparticles with alternating hydrophilic–hydrophobic–hydrophilic domains into polymer membranes does not occur as a thermally activated event. PMID:26029770

  7. Preparation of polysaccharide loaded collagen membrane with anti-oxidative activity.

    PubMed

    Shu, Zibin; Ding, Shengli; He, Xiaohong; Dai, Xuemei; Xiao, Qian; Yang, Min; Leng, Xue; Ma, Yanshun; Yang, Hua

    2015-01-01

    The scavenging activity of polysaccharides from Lycium barbarum, Lentinus edodes and Ganoderma Lucidum Karst to DPPH free radicals was investigated. It was found that among the three polysaccharides, Lycium barbarum polysaccharides (LBP) exhibits the best scavenging activity. Polysaccharide loaded collagen membranes were prepared by mixing LBP with collagen, starch, glycerol, sodium carboxymethyl cellulose and glutaraldehyde. In vitro drug release from membranes was evaluated. With increasing the immersion time, the release rate first increases and then slows down. Meanwhile, the scavenging activity to DPPH radicals exhibits similar variation, in agreement with a good release effect of the membrane. The optimal formulation of collagen membrane and preparation parameters were obtained considering the overall properties and the scavenging activity to radicals. PMID:26406078

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  9. Gβ1γ2 activates phospholipase A2-dependent Golgi membrane tubule formation

    PubMed Central

    Bechler, Marie E.; Brown, William J.

    2014-01-01

    Heterotrimeric G proteins transduce the ligand binding of transmembrane G protein coupled receptors into a variety of intracellular signaling pathways. Recently, heterotrimeric Gβγ subunit signaling at the Golgi complex has been shown to regulate the formation of vesicular transport carriers that deliver cargo from the Golgi to the plasma membrane. In addition to vesicles, membrane tubules have also been shown to mediate export from the Golgi complex, which requires the activity of cytoplasmic phospholipase A2 (PLA2) enzyme activity. Through the use of an in vitro reconstitution assay with isolated Golgi complexes, we provide evidence that Gβ1γ2 signaling also stimulates Golgi membrane tubule formation. In addition, we show that an inhibitor of Gβγ activation of PLA2 enzymes inhibits in vitro Golgi membrane tubule formation. Additionally, purified Gβγ protein stimulates membrane tubules in the presence of low (sub-threshold) cytosol concentrations. Importantly, this Gβγ stimulation of Golgi membrane tubule formation was inhibited by treatment with the PLA2 antagonist ONO-RS-082. These studies indicate that Gβ1γ2 signaling activates PLA2 enzymes required for Golgi membrane tubule formation, thus establishing a new layer of regulation for this process. PMID:25019068

  10. cBid, Bax and Bcl-xL exhibit opposite membrane remodeling activities

    PubMed Central

    Bleicken, S; Hofhaus, G; Ugarte-Uribe, B; Schröder, R; García-Sáez, A J

    2016-01-01

    The proteins of the Bcl-2 family have a crucial role in mitochondrial outer membrane permeabilization during apoptosis and in the regulation of mitochondrial dynamics. Current models consider that Bax forms toroidal pores at mitochondria that are responsible for the release of cytochrome c, whereas Bcl-xL inhibits pore formation. However, how Bcl-2 proteins regulate mitochondrial fission and fusion remains poorly understood. By using a systematic analysis at the single vesicle level, we found that cBid, Bax and Bcl-xL are able to remodel membranes in different ways. cBid and Bax induced a reduction in vesicle size likely related to membrane tethering, budding and fission, besides membrane permeabilization. Moreover, they are preferentially located at highly curved membranes. In contrast, Bcl-xL not only counterbalanced pore formation but also membrane budding and fission. Our findings support a mechanism of action by which cBid and Bax induce or stabilize highly curved membranes including non-lamellar structures. This molecular activity reduces the energy for membrane remodeling, which is a necessary step in toroidal pore formation, as well as membrane fission and fusion, and provides a common mechanism that links the two main functions of Bcl-2 proteins. PMID:26913610

  11. cBid, Bax and Bcl-xL exhibit opposite membrane remodeling activities.

    PubMed

    Bleicken, S; Hofhaus, G; Ugarte-Uribe, B; Schröder, R; García-Sáez, A J

    2016-01-01

    The proteins of the Bcl-2 family have a crucial role in mitochondrial outer membrane permeabilization during apoptosis and in the regulation of mitochondrial dynamics. Current models consider that Bax forms toroidal pores at mitochondria that are responsible for the release of cytochrome c, whereas Bcl-xL inhibits pore formation. However, how Bcl-2 proteins regulate mitochondrial fission and fusion remains poorly understood. By using a systematic analysis at the single vesicle level, we found that cBid, Bax and Bcl-xL are able to remodel membranes in different ways. cBid and Bax induced a reduction in vesicle size likely related to membrane tethering, budding and fission, besides membrane permeabilization. Moreover, they are preferentially located at highly curved membranes. In contrast, Bcl-xL not only counterbalanced pore formation but also membrane budding and fission. Our findings support a mechanism of action by which cBid and Bax induce or stabilize highly curved membranes including non-lamellar structures. This molecular activity reduces the energy for membrane remodeling, which is a necessary step in toroidal pore formation, as well as membrane fission and fusion, and provides a common mechanism that links the two main functions of Bcl-2 proteins. PMID:26913610

  12. Daptomycin exerts rapid bactericidal activity against Bacillus anthracis without disrupting membrane integrity

    PubMed Central

    Xing, Yu-hua; Wang, Wei; Dai, Su-qin; Liu, Ti-yan; Tan, Jun-jie; Qu, Guo-long; Li, Yu-xia; Ling, Yan; Liu, Gang; Fu, Xue-qi; Chen, Hui-peng

    2014-01-01

    Aim: To examine whether the novel cyclic lipopeptide antibiotic daptomycin could be used to treat anthrax and to study the mechanisms underlying its bactericidal action against Bacillus anthracis. Methods: Spore-forming B anthracis AP422 was tested. MIC values of antibiotics were determined. Cell membrane potential was measured using flow cytometric assays with membrane potential-sensitive fluorescent dyes. Cell membrane integrity was detected using To-Pro-3 iodide staining and transmission electron microscopy. K+ efflux and Na+ influx were measured using the fluorescent probes PBFI and SBFI-AM, respectively. Results: Daptomycin exhibited rapid bactericidal activity against vegetative B anthracis with a MIC value of 0.78 μg/mL, which was comparable to those of ciprofloxacin and penicillin G. Furthermore, daptomycin prevented the germinated spores from growing into vegetative bacteria. Daptomycin concentration-dependently dissipated the membrane potential of B anthracis and caused K+ efflux and Na+ influx without disrupting membrane integrity. In contrast, both ciprofloxacin and penicillin G did not change the membrane potential of vegetative bacteria or spores. Penicillin G disrupted membrane integrity of B anthracis, whereas ciprofloxacin had no such effect. Conclusion: Daptomycin exerts rapid bactericidal action against B anthracis via reducing membrane potential without disrupting membrane integrity. This antibiotic can be used as an alternate therapy for B anthracis infections. PMID:24362329

  13. Dissociation of membrane binding and lytic activities of the lymphocyte pore-forming protein (perforin).

    PubMed

    Young, J D; Damiano, A; DiNome, M A; Leong, L G; Cohn, Z A

    1987-05-01

    Granules isolated from CTL and NK cells contain a cytolytic pore-forming protein (PFP/perforin). At low temperatures (on ice), PFP binds to erythrocyte membranes without producing hemolysis. Hemolysis occurs when the PFP-bound erythrocytes are warmed up to 37 degrees C, which defines a temperature-dependent, lytic (pore-formation) step distinct from the membrane-binding event. Ca2+ and neutral pH are required for both membrane binding and pore formation by PFP. Serum, LDL, HDL, and heparin inhibit the hemolytic activity of PFP by blocking its binding to lipid membranes. Lysis by PFP that has bound to erythrocyte membranes is no longer susceptible to the effect of these inhibitors. The hemolytic activities associated with intact granules and solubilized PFP show different requirements for Ca2+ and pH, indicating that cytolysis produced by isolated granules may involve an additional step, possibly fusion of granules with membranes. It is suggested that three distinct Ca2+- and pH-dependent events may be involved during cell killing by CTL and NK cells: fusion of cytoplasmic granules of effector cells with their plasma membrane, releasing PFP from cells; binding of the released PFP to target membranes; and insertion of monomers and the subsequent formation of lytic pores in the target membrane. The serum-mediated inhibition of membrane binding by PFP could prevent the accidental injury of bystander cells by cell-released PFP, but would allow cytolysis to proceed to completion once PFP has bound to the target membrane. PMID:3494808

  14. High-pressure stainless steel active membrane microvalves

    NASA Astrophysics Data System (ADS)

    Sharma, G.; Svensson, S.; Ogden, S.; Klintberg, L.; Hjort, K.

    2011-07-01

    In this work, high-pressure membrane microvalves have been designed, manufactured and evaluated. The valves were able to withstand back-pressures of 200 bar with a response time of less than 0.6 s. These stainless steel valves, manufactured with back-end batch production, utilize the large volume expansion coupled to the solid-liquid phase transition in paraffin wax. When membrane materials were evaluated, parylene coated stainless steel was found to be the best choice as compared to polydimethylsiloxane and polyimide. Also, the influence of the orifice placement and diameter is included in this work. If the orifice is placed too close to the rim of the membrane, the valve can stay sealed even after turning the power off, and the valve will not open until the pressure in the system is released. The developed steel valves, evaluated for both water and air, provide excellent properties in terms of mechanical stability, ease of fabrication, and low cost. Possible applications include sampling at high pressures, chemical microreactors, high performance liquid chromatography, pneumatics, and hydraulics.

  15. Large-aperture fast multilevel Fresnel zone lenses in glass and ultrathin polymer films for visible and near-infrared imaging applications.

    PubMed

    Britten, Jerald A; Dixit, Shamusundar N; DeBruyckere, Michael; Steadfast, Daniel; Hackett, James; Farmer, Brandon; Poe, Garrett; Patrick, Brian; Atcheson, Paul D; Domber, Jeanette L; Seltzer, Aaron

    2014-04-10

    The ability to fabricate 4-level diffractive structures with 1 µm critical dimensions has been demonstrated for the creation of fast (∼f/3.1 at 633 nm) Fresnel zone lenses (FZLs) with >60% diffraction efficiency into the -1 focusing order and nearly complete suppression of 0 and +1 orders. This is done using tooling capable of producing optics with 800 mm apertures. A 4-level grating fabricated in glass at 300 mm aperture is shown to have <15  nm rms holographic phase error. Glass FZLs have also been used as mandrels for casting zero-thermal-expansion, 20 µm thick polymer films created with the 4-level structure as a route to mass replication of efficient diffractive membranes for ultralight segmented space-based telescope applications. PMID:24787399

  16. Estimating the magnitude of near-membrane PDE4 activity in living cells.

    PubMed

    Xin, Wenkuan; Feinstein, Wei P; Britain, Andrea L; Ochoa, Cristhiaan D; Zhu, Bing; Richter, Wito; Leavesley, Silas J; Rich, Thomas C

    2015-09-15

    Recent studies have demonstrated that functionally discrete pools of phosphodiesterase (PDE) activity regulate distinct cellular functions. While the importance of localized pools of enzyme activity has become apparent, few studies have estimated enzyme activity within discrete subcellular compartments. Here we present an approach to estimate near-membrane PDE activity. First, total PDE activity is measured using traditional PDE activity assays. Second, known cAMP concentrations are dialyzed into single cells and the spatial spread of cAMP is monitored using cyclic nucleotide-gated channels. Third, mathematical models are used to estimate the spatial distribution of PDE activity within cells. Using this three-tiered approach, we observed two pharmacologically distinct pools of PDE activity, a rolipram-sensitive pool and an 8-methoxymethyl IBMX (8MM-IBMX)-sensitive pool. We observed that the rolipram-sensitive PDE (PDE4) was primarily responsible for cAMP hydrolysis near the plasma membrane. Finally, we observed that PDE4 was capable of blunting cAMP levels near the plasma membrane even when 100 μM cAMP were introduced into the cell via a patch pipette. Two compartment models predict that PDE activity near the plasma membrane, near cyclic nucleotide-gated channels, was significantly lower than total cellular PDE activity and that a slow spatial spread of cAMP allowed PDE activity to effectively hydrolyze near-membrane cAMP. These results imply that cAMP levels near the plasma membrane are distinct from those in other subcellular compartments; PDE activity is not uniform within cells; and localized pools of AC and PDE activities are responsible for controlling cAMP levels within distinct subcellular compartments. PMID:26201952

  17. Opposite effect of membrane raft perturbation on transport activity of KCC2 and NKCC1.

    PubMed

    Hartmann, Anna-Maria; Blaesse, Peter; Kranz, Thorsten; Wenz, Meike; Schindler, Jens; Kaila, Kai; Friauf, Eckhard; Nothwang, Hans Gerd

    2009-10-01

    In the majority of neurons, the intracellular Cl(-) concentration is set by the activity of the Na(+)-K(+)-2Cl(-) cotransporter (NKCC1) and the K(+)-Cl(-) cotransporter (KCC2). Here, we investigated the cotransporters' functional dependence on membrane rafts. In the mature rat brain, NKCC1 was mainly insoluble in Brij 58 and co-distributed with the membrane raft marker flotillin-1 in sucrose density flotation experiments. In contrast, KCC2 was found in the insoluble fraction as well as in the soluble fraction, where it co-distributed with the non-raft marker transferrin receptor. Both KCC2 populations displayed a mature glycosylation pattern. Disrupting membrane rafts with methyl-beta-cyclodextrin (MbetaCD) increased the solubility of KCC2, yet had no effect on NKCC1. In human embryonic kidney-293 cells, KCC2 was strongly activated by a combined treatment with MbetaCD and sphingomyelinase, while NKCC1 was inhibited. These data indicate that membrane rafts render KCC2 inactive and NKCC1 active. In agreement with this, inactive KCC2 of the perinatal rat brainstem largely partitioned into membrane rafts. In addition, the exposure of the transporters to MbetaCD and sphingomyelinase showed that the two transporters differentially interact with the membrane rafts. Taken together, membrane raft association appears to represent a mechanism for co-ordinated regulation of chloride transporter function. PMID:19686239

  18. Fusion-Triggered Switching of Enzymatic Activity on an Artificial Cell Membrane

    PubMed Central

    Mukai, Masaru; Sasaki, Yoshihiro; Kikuchi, Jun-ichi

    2012-01-01

    A nanosensory membrane device was constructed for detecting liposome fusion through changes in an enzymatic activity. Inspired by a biological signal transduction system, the device design involved functionalized liposomal membranes prepared by self-assembly of the following molecular components: a synthetic peptide lipid and a phospholipid as matrix membrane components, a Schiff's base of pyridoxal 5′-phosphate with phosphatidylethanolamine as a thermo-responsive artificial receptor, NADH-dependent L-lactate dehydrogenase as a signal amplifier, and Cu2+ ion as a signal mediator between the receptor and enzyme. The enzymatic activity of the membrane device was adjustable by changing the matrix lipid composition, reflecting the thermotropic phase transition behavior of the lipid membranes, which in turn controlled receptor binding affinity toward the enzyme-inhibiting mediator species. When an effective fusogen anionic polymer was added to these cationic liposomes, membrane fusion occurred, and the functionalized liposomal membranes responded with changes in enzymatic activity, thus serving as an effective nanosensory device for liposome fusion detection. PMID:22778625

  19. Characterization of a secretase activity which releases angiotensin-converting enzyme from the membrane.

    PubMed Central

    Oppong, S Y; Hooper, N M

    1993-01-01

    Angiotensin-converting enzyme (ACE; EC 3.4.1.15.1) exists in both membrane-bound and soluble forms. Phase separation in Triton X-114 and a competitive e.l.i.s.a. have been employed to characterize the activity which post-translationally converts the amphipathic, membrane-bound form of ACE in pig kidney microvilli into a hydrophilic, soluble form. This secretase activity was enriched to a similar extent as other microvillar membrane proteins, was tightly membrane-associated, being resistant to extensive washing of the microvillar membranes with 0.5 M NaCl, and displayed a pH optimum of 8.4. The ACE secretase was not affected by inhibitors of serine-, thiol- or aspartic-proteases, nor by reducing agents or alpha 2-macroglobulin. The metal chelators, EDTA and 1,10-phenanthroline, inhibited the secretase activity, with, in the case of EDTA, an inhibitor concentration of 2.5 mM causing 50% inhibition. In contrast, EGTA inhibited the secretase by a maximum of 15% at a concentration of 10 mM. The inhibition of EDTA was reactivated substantially (83%) by Mg2+ ions, and partially (34% and 29%) by Zn2+ and Mn2+ ions respectively. This EDTA-sensitive secretase activity was also present in microsomal membranes prepared from pig lung and testis, and from human lung and placenta, but was absent from human kidney and human and pig intestinal brush-border membranes. The form of ACE released from the microvillar membrane by the secretase co-migrated on SDS/PAGE with ACE purified from pig plasma, thus the action and location of the secretase would be consistent with it possibly having a role in the post-translational proteolytic cleavage of membrane-bound ACE to generate the soluble form found in blood, amniotic fluid, seminal plasma and other body fluids. Images Figure 4 PMID:8389141

  20. Membrane activity of the phospholipase C-δ1 pleckstrin homology (PH) domain

    PubMed Central

    2005-01-01

    PH-PLCδ1 [the PH domain (pleckstrin homology domain) of PLCδ1 (phospholipase C-δ1)] is among the best-characterized phosphoinositide-binding domains. PH-PLCδ1 binds with high specificity to the headgroup of PtdIns(4,5)P2, but little is known about its interfacial properties. In the present study, we show that PH-PLCδ1 is also membrane-active and can insert significantly into PtdIns(4,5)P2-containing monolayers at physiological (bilayer-equivalent) surface pressures. However, this membrane activity appears to involve interactions distinct from those that target PH-PLCδ1 to the PtdIns(4,5)P2 headgroup. Whereas the majority of PtdIns(4,5)P2-bound PH-PLCδ1 can be displaced by adding excess of soluble headgroup [Ins(1,4,5)P3], membrane activity of PH-PLCδ1 cannot. PH-PLCδ1 differs from other phosphoinositide-binding domains in that its membrane insertion does not require that the phosphoinositide-binding site be occupied. Significant monolayer insertion remains when the phosphoinositide-binding site is mutated, and PH-PLCδ1 can insert into monolayers that contain no PtdIns(4,5)P2 at all. Our results suggest a model in which reversible membrane binding of PH-PLCδ1, mediated by PtdIns(4,5)P2 or other acidic phospholipids, occurs without membrane insertion. Accumulation of the PH domain at the membrane surface enhances the efficiency of insertion, but does not significantly affect its extent, whereas the presence of phosphatidylethanolamine and cholesterol in the lipid mixture promotes the extent of insertion. This is the first report of membrane activity in an isolated PH domain and has implications for understanding the membrane targeting by this common type of domain. PMID:15755258

  1. Neuronal Activation by GPI-Linked Neuroligin-1 Displayed in Synthetic Lipid Bilayer Membranes

    PubMed Central

    Baksh, Michael M.; Groves, Jay T.; Dean, Camin; Pautot, Sophie; DeMaria, Shannon; Isacoff, Ehud

    2006-01-01

    We have characterized, in vitro, interactions between hippocampal neuronal cells and silica microbeads coated with synthetic, fluid, lipid bilayer membranes containing the glycosylphosphatidyl inositol (GPI)-linked extracellular domain of the postsynaptic membrane protein neuroligin-1. These bilayer–neuroligin-1 beads activated neuronal cells to form presynaptic nerve terminals at the point of contact in a manner similar to that observed for live PC12 cells, ectopically expressing the full length neuroligin-1. The synthetic membranes exhibited biological activity at neuroligin-1 densities of ∼1 to 6 proteins/μm2. Polyolycarbonate beads with neuroligin-1 covalently attached to the surface failed to activate neurons despite the fact that neuroligin-1 binding activity is preserved. This implies that a lipid membrane environment is likely to be essential for neuroligin-1 activity. This technique allows the study of isolated proteins in an environment that has physical properties resembling those of a cell surface; proteins can diffuse freely within the membrane, retain their in vivo orientations, and are in a nondenatured state. In addition, the synthetic membrane environment affords control over both lipid and protein composition. This technology is easily implemented and can be applied to a wide variety of cellular studies. PMID:16262338

  2. Sodium pump molecular activity and membrane lipid composition in two disparate ectotherms, and comparison with endotherms.

    PubMed

    Turner, Nigel; Hulbert, A J; Else, Paul L

    2005-02-01

    Previous research has shown that the lower sodium pump molecular activity observed in tissues of ectotherms compared to endotherms, is largely related to the lower levels of polyunsaturates and higher levels of monounsaturates found in the cell membranes of ectotherms. Marine-based ectotherms, however, have very polyunsaturated membranes, and in the current study, we measured molecular activity and membrane lipid composition in tissues of two disparate ectothermic species, the octopus (Octopus vulgaris) and the bearded dragon lizard (Pogona vitticeps), to determine whether the high level of membrane polyunsaturation generally observed in marine-based ectotherms is associated with an increased sodium pump molecular activity relative to other ectotherms. Phospholipids from all tissues of the octopus were highly polyunsaturated and contained high concentrations of the omega-3 polyunsaturate, docosahexaenoic acid (22:6 (n-3)). In contrast, phospholipids from bearded dragon tissues contained higher proportions of monounsaturates and lower proportions of polyunsaturates. Sodium pump molecular activity was only moderately elevated in tissues of the octopus compared to the bearded dragon, despite the much greater level of polyunsaturation in octopus membranes. When the current data were combined with data for the ectothermic cane toad, a significant (P = 0.003) correlation was observed between sodium pump molecular activity and the content of 22:6 (n-3) in the surrounding membrane. These results are discussed in relation to recent work which shows a similar relationship in endotherms. PMID:15726386

  3. Investigation of the pore structure and morphology of cellulose acetate membranes using small-angle neutron scattering. 1: Cellulose acetate active layer membranes

    SciTech Connect

    Kulkarni, S.; Krause, S. ); Wignall, G.D. . Solid State Div.); Hammouda, B. . Center for High Resolution Neutron Scattering)

    1994-11-07

    The structure of ultrathin cellulose acetate membranes, known as active layer membranes, has been investigated using small-angle neutron scattering. These membranes are known to have structural and functional similarity to the surface or skin layer in commercial reverse-osmosis (RO) membranes and hence are useful model systems for understanding the structure of the RO membrane skin layer. Active layer membranes were studied after swelling them with either D[sub 2]O or CD[sub 3]OD. The results in both cases clearly indicated the presence of very small (10--20 [angstrom]) porous structures in the membrane. The presence of such pores has been a subject of long-standing controversy in this area. The data were analyzed using a modified Debye-Bueche analysis and the resultant membrane structure was seen to agree well with structural information from electron microscopic studies. Finally, a possible explanation for the differences in scattering observed between the D[sub 2]O swollen membranes and the CD[sub 3]OD swollen membranes has been presented.

  4. Membrane permeability and the loss of germination factor from Neurospora crassa at low water activities

    NASA Technical Reports Server (NTRS)

    Charlang, G.; Horowitz, N. H.

    1974-01-01

    Neurospora crassa conidia incubating in buffer at low water activities release a germination-essential component as well as 260-nm absorbing and ninhydrin-positive materials, regardless of whether an electrolyte or nonelectrolyte is used to reduce water activity. Chloroform and antibiotics known to increase cell-membrane permeability have a similar effect. This suggests that membrane damage occurs in media of low water activity and that an increase in permeability is responsible for the release of cellular components. The damage caused in media of low water activity is nonlethal in most cases, and the conidia recover when transferred to nutrient medium.

  5. The antifungal activity and membrane-disruptive action of dioscin extracted from Dioscorea nipponica.

    PubMed

    Cho, Jaeyong; Choi, Hyemin; Lee, Juneyoung; Kim, Mi-Sun; Sohn, Ho-Yong; Lee, Dong Gun

    2013-03-01

    Dioscin is a kind of steroidal saponin isolated from the root bark of wild yam Dioscorea nipponica. We investigated the antifungal effect of dioscin against different fungal strains and its antifungal mechanism(s) in Candida albicans cells. Using the propidium iodide assay and calcein-leakage measurement, we confirmed that dioscin caused fungal membrane damage. Furthermore, we evaluated the ability of dioscin to disrupt the plasma membrane potential, using 3,3'-dipropylthiadicarbocyanine iodide [DiSC(3)(5)] and bis-(1,3-dibarbituric acid)-trimethine oxanol [DiBAC(4)(3)]. Cells stained with the dyes had a significant increase in fluorescent intensity after exposure to dioscin, indicating that dioscin has an effect on the membrane potential. To visualize the effect of dioscin on the cell membrane, we synthesized rhodamine-labeled giant unilamellar vesicles (GUVs) mimicking the outer leaflet of the plasma membrane of C. albicans. As seen in the result, the membrane disruptive action of dioscin caused morphological change and rhodamine leakage of the GUVs. In three-dimensional contour-plot analysis using flow cytometry, we observed a decrease in cell size, which is in agreement with our result from the GUV assay. These results suggest that dioscin exerts a considerable antifungal activity by disrupting the structure in membrane after invading into the fungal membrane, resulting in fungal cell death. PMID:23262192

  6. Powder Activated Carbon Pretreatment of a Microfiltration Membrane for the Treatment of Surface Water

    PubMed Central

    Song, Yali; Dong, Bingzhi; Gao, Naiyun; Ma, Xiaoyan

    2015-01-01

    This study focused on the effect of powder activated carbon (PAC) adsorption on microfiltration (MF) membrane performance. The results showed that PAC pretreatment offered high organic matter removal rates for both dissolved organic carbon (DOC) and ultraviolet absorbance at 254 nm (UV254) during 10–200 mg/L PAC dosage. The removal efficiencies of organic matter by MF membrane filtration decreased with the increase of organic matter removal rate by PAC adsorption. PAC mainly removed organic matter of about 3 kDa molecular weight (MW). MF membrane maintained more than 5 kDa MW organic matter on the membrane after PAC adsorption. The results of membrane filtration indicated that PAC pretreatment slightly promoted membrane flux, regardless of PAC dosage. It seems that the organic matter fouling membrane was concentrated in more than 3 kDa MW. PAC removed markedly less than 3 kDa MW organic matter and had less effect on more than 3 kDa organic matter. Thus, PAC cannot reduce membrane fouling. PMID:26378552

  7. Characterization of antibacterial polyethersulfone membranes using the Respiration Activity Monitoring System (RAMOS).

    PubMed

    Kochan, Jozef; Scheidle, Marco; van Erkel, Joost; Bikel, Matías; Büchs, Jochen; Wong, John Erik; Melin, Thomas; Wessling, Matthias

    2012-10-15

    Membranes with antibacterial properties were developed using surface modification of polyethersulfone ultrafiltration membranes. Three different modification strategies using polyelectrolyte layer-by-layer (LbL) technique are described. The first strategy relying on the intrinsic antibacterial properties of poly(diallyldimethylammonium chloride) (PDADMAC) and poly(ethylenimine) (PEI) exhibits only little antibacterial effects. The other two strategies contain silver in both ionic (Ag(+)) and metallic (Ag(0)) form. Ag(+) embedded into negatively charged poly(sodium 4-styrene sulfonate) (PSS) layers totally inhibits bacterial growth. Ag(0) nanoparticles were introduced to the membrane surface by LbL deposition of chitosan- and poly(methacrylic acid) - sodium salt (PMA)-capped silver nanoparticles and subsequent UV or heat treatment. Antibacterial properties of the modified membranes were quantified by a new method based on the Respiration Activity Monitoring System (RAMOS), whereby the oxygen transfer rates (OTR) of E. coli K12 cultures on the membranes were monitored online. As opposed to colony forming counting method RAMOS yields more quantitative and reliable data on the antibacterial effect of membrane modification. Ag-imprinted polyelectrolyte film composed of chitosan (Ag(0))/PMA(Ag(0))/chitosan(Ag(0)) was found to be the most promising among the tested membranes. Further investigation revealed that the concentration and equal distribution of silver in the membrane surface plays an important role in bacterial growth inhibition. PMID:22884245

  8. Membrane-Active Sequences within gp41 Membrane Proximal External Region (MPER) Modulate MPER-Containing Peptidyl Fusion Inhibitor Activity and the Biosynthesis of HIV-1 Structural Proteins

    PubMed Central

    Zhang, Si Min; Jejcic, Alenka; Tam, James P.; Vahlne, Anders

    2015-01-01

    The membrane proximal external region (MPER) is a highly conserved membrane-active region located at the juxtamembrane positions within class I viral fusion glycoproteins and essential for membrane fusion events during viral entry. The MPER in the human immunodeficiency virus type I (HIV-1) envelope protein (Env) interacts with the lipid bilayers through a cluster of tryptophan (Trp) residues and a C-terminal cholesterol-interacting motif. The inclusion of the MPER N-terminal sequence contributes to the membrane reactivity and anti-viral efficacy of the first two anti-HIV peptidyl fusion inhibitors T20 and T1249. As a type I transmembrane protein, Env also interacts with the cellular membranes during its biosynthesis and trafficking. Here we investigated the roles of MPER membrane-active sequences during both viral entry and assembly, specifically, their roles in the design of peptidyl fusion inhibitors and the biosynthesis of viral structural proteins. We found that elimination of the membrane-active elements in MPER peptides, namely, penta Trp→alanine (Ala) substitutions and the disruption of the C-terminal cholesterol-interacting motif through deletion inhibited the anti-viral effect against the pseudotyped HIV-1. Furthermore, as compared to C-terminal dimerization, N-terminal dimerization of MPER peptides and N-terminal extension with five helix-forming residues enhanced their anti-viral efficacy substantially. The secondary structure study revealed that the penta-Trp→Ala substitutions also increased the helical content in the MPER sequence, which prompted us to study the biological relevance of such mutations in pre-fusion Env. We observed that Ala mutations of Trp664, Trp668 and Trp670 in MPER moderately lowered the intracellular and intraviral contents of Env while significantly elevating the content of another viral structural protein, p55/Gag and its derivative p24/capsid. The data suggest a role of the gp41 MPER in the membrane-reactive events during

  9. The role of spontaneous lipid curvature in the interaction of interfacially active peptides with membranes.

    PubMed

    Koller, Daniel; Lohner, Karl

    2014-09-01

    Research on antimicrobial peptides is in part driven by urgent medical needs such as the steady increase in pathogens being resistant to antibiotics. Despite the wealth of information compelling structure-function relationships are still scarce and thus the interfacial activity model has been proposed to bridge this gap. This model also applies to other interfacially active (membrane active) peptides such as cytolytic, cell penetrating or antitumor peptides. One parameter that is strongly linked to interfacial activity is the spontaneous lipid curvature, which is experimentally directly accessible. We discuss different parameters such as H-bonding, electrostatic repulsion, changes in monolayer surface area and lateral pressure that affect induction of membrane curvature, but also vice versa how membrane curvature triggers peptide response. In addition, the impact of membrane lipid composition on the formation of curved membrane structures and its relevance for diverse mode of action of interfacially active peptides and in turn biological activity are described. This article is part of a Special Issue entitled: Interfacially Active Peptides and Proteins. Guest Editors: William C. Wimley and Kalina Hristova. PMID:24853655

  10. Phosphatidylinositol kinase is activated in membranes derived from cells treated with epidermal growth factor.

    PubMed Central

    Walker, D H; Pike, L J

    1987-01-01

    The ability of epidermal growth factor (EGF) to stimulate phosphatidylinositol (PtdIns) kinase activity in A431 cells was examined. The incorporation of 32P from [gamma-32P]ATP into PtdIns by A431 membranes was increased in membranes prepared from cells that had been pretreated with EGF. Demonstration of a stimulation of the PtdIns kinase activity by EGF required the use of subconfluent cultures and was dependent on the inclusion of protease inhibitors in the buffers used to prepare the membranes. Stimulation of the PtdIns kinase activity was rapid. The activation peaked 2 min after the addition of EGF and declined slowly thereafter. Half-maximal stimulation of the PtdIns kinase occurred at 7 nM EGF. Kinetic analyses of the reaction indicated that treatment of the cells with EGF resulted in a decrease in the Km for PtdIns with no change in the Vmax. The kinetic parameters for the utilization of ATP were unchanged in the EGF-treated membranes compared to the control membranes. Pretreatment of the cells with the phorbol ester phorbol 12-myristate 13-acetate blocked the ability of EGF to stimulate PtdIns kinase activity. These findings demonstrate that a PtdIns kinase activity in A431 cells is regulated by EGF and provide a good system for examining the mechanism by which EGF stimulates the activity of this intracellular enzyme. PMID:2823265

  11. Regulation of membrane associated protein kinase C activity by guanine nucleotide in rabbit peritoneal neutrophils

    SciTech Connect

    Huang, C.K.; Devanney, J.F.

    1986-03-05

    Addition of phorbol myristate acetate (PMA) (0.1 ..mu..g/ml) or guanosine-5'-0-(3-thiotriphosphate) (GTP..gamma..S) (10..mu..M) to the membrane fraction from rabbit peritoneal neutrophils results in an increase of phosphorylation of several membrane proteins. To test whether membrane associated protein kinase C is involved in the activation, histone is added to the membrane as a substrate for protein kinase C. Phosphorylation of histone is determined by counting the gel pieces containing histone IIIS after separation from other membrane components by SDS-gel electrophoresis. In the presence of CaC12 (20 ..mu..M), GTP..gamma..S (10 ..mu..M) or PMA (0.1 ..mu..g/ml) stimulates the phosphorylation of histone IIIS (40% to 70% increase). To achieve this effect calcium is required for GTP..gamma..S but not for PMA. The effect of GTP..gamma..S but not PMA is inhibited in membranes obtained from cells pretreated with pertussis toxin. Membrane protein kinase C is solubilized with Triton X-100 (1%) and then applied to a DEAE-52 cellulose column chromatography. Two peaks of protein kinase C activity are observed. Peak one is eluted at 40 mM NaCl, peak two is eluted at 140 mM NaCl. The activity of peak one is stimulated with phosphatidylserine (PS) and PMA but not with PS and calcium. The activity of peak two is stimulated with either PS and PMA or PS and calcium. The results suggest that GTP binding protein is involved in the activation of membrane associated protein kinase C and the kinase may exist in two forms, calcium sensitive and calcium insensitive.

  12. Activity of immobilised rat hepatic microsomal CYP2E1 using alumina membrane as a support.

    PubMed

    Tanvir, Shazia; Morandat, Sandrine; Frederic, Nadaud; Adenier, Hervé; Pulvin, Sylviane

    2009-11-30

    Porous alumina membranes are attractive materials for the construction of biosensors and also have utility for the production of immobilised enzyme bioreactors. Microsomes from rat liver were adsorbed onto alumina membrane activated by silane. Microsomal membranes were pumped through the channels where they became immobilised by binding to amine groups on the surface of the alumina membrane. In an effort to gain a quantitative understanding of the effects of microsomal film growth on enzyme activity, we compared the para-nitrophenol (pNP) hydroxylase activity of the microsomes by varying the amount of microsomes fixed in alumina microchannels. The alumina membrane was placed in a fluidic device at a fast flow that afforded short residence time (seconds) to obtain transformation of pNP to 4-nitrocatechol (pNC), which was detected by LC-MS/MS. This enabled the use of this bioreactor where CYP2E1 activity is low and tissue sources are limiting. The microsomes, successfully immobilised on the alumina membranes, were used to produce stable biocatalytic reactors that can be used repeatedly over a period of 2 months. PMID:19703600

  13. G-1-activated membrane estrogen receptors mediate increased contractility of the human myometrium.

    PubMed

    Maiti, K; Paul, J W; Read, M; Chan, E C; Riley, S C; Nahar, P; Smith, R

    2011-06-01

    Estrogens are key mediators of increased uterine contractility at labor. We sought to determine whether membrane-associated estrogen receptors, such as the recently described seven-transmembrane receptor G protein-coupled receptor 30 (GPR30), mediated some of this effect. Using human myometrium obtained at term cesarean section before or after the onset of labor, we demonstrated the presence of GPR30 mRNA and protein using quantitative RT-PCR and Western blotting. GPR30 receptor was localized to the cell membrane and often colocalized with calveolin-1. Using the specific estrogen membrane receptor agonist G-1 and myometrial explants, we showed that membrane receptor activation led to phosphorylation of MAPK and the actin-modifying small heat shock protein 27. Using myometrial strips incubated with G-1 or vehicle we demonstrated that estrogen membrane receptor activation increased the myometrial contractile response to oxytocin. These data suggest that activation of the plasma membrane estrogen receptor GPR30 likely participates in the physiology of the human myometrium during pregnancy and identifies it as a potential target to modify uterine activity. PMID:21427217

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

    PubMed Central

    Tran, Jason; Magenau, Astrid; Rodriguez, Macarena; Rentero, Carles; Royo, Teresa; Enrich, Carlos; Thomas, Shane R.; Grewal, Thomas; Gaus, Katharina

    2016-01-01

    Endothelial cells respond to a large range of stimuli including circulating lipoproteins, growth factors and changes in haemodynamic mechanical forces to regulate the activity of endothelial nitric oxide synthase (eNOS) and maintain blood pressure. While many signalling pathways have been mapped, the identities of membrane domains through which these signals are transmitted are less well characterized. Here, we manipulated bovine aortic endothelial cells (BAEC) with cholesterol and the oxysterol 7-ketocholesterol (7KC). Using a range of microscopy techniques including confocal, 2-photon, super-resolution and electron microscopy, we found that sterol enrichment had differential effects on eNOS and caveolin-1 (Cav1) colocalisation, membrane order of the plasma membrane, caveolae numbers and Cav1 clustering. We found a correlation between cholesterol-induced condensation of the plasma membrane and enhanced high density lipoprotein (HDL)-induced eNOS activity and phosphorylation suggesting that cholesterol domains, but not individual caveolae, mediate HDL stimulation of eNOS. Vascular endothelial growth factor (VEGF)-induced and shear stress-induced eNOS activity was relatively independent of membrane order and may be predominantly controlled by the number of caveolae on the cell surface. Taken together, our data suggest that signals that activate and phosphorylate eNOS are transmitted through distinct membrane domains in endothelial cells. PMID:26977592

  15. Structural basis for activation, assembly and membrane binding of ESCRT-III Snf7 filaments

    PubMed Central

    Tang, Shaogeng; Henne, W Mike; Borbat, Peter P; Buchkovich, Nicholas J; Freed, Jack H; Mao, Yuxin; Fromme, J Christopher; Emr, Scott D

    2015-01-01

    The endosomal sorting complexes required for transport (ESCRTs) constitute hetero-oligomeric machines that catalyze multiple topologically similar membrane-remodeling processes. Although ESCRT-III subunits polymerize into spirals, how individual ESCRT-III subunits are activated and assembled together into a membrane-deforming filament remains unknown. Here, we determine X-ray crystal structures of the most abundant ESCRT-III subunit Snf7 in its active conformation. Using pulsed dipolar electron spin resonance spectroscopy (PDS), we show that Snf7 activation requires a prominent conformational rearrangement to expose protein-membrane and protein-protein interfaces. This promotes the assembly of Snf7 arrays with ~30 Å periodicity into a membrane-sculpting filament. Using a combination of biochemical and genetic approaches, both in vitro and in vivo, we demonstrate that mutations on these protein interfaces halt Snf7 assembly and block ESCRT function. The architecture of the activated and membrane-bound Snf7 polymer provides crucial insights into the spatially unique ESCRT-III-mediated membrane remodeling. DOI: http://dx.doi.org/10.7554/eLife.12548.001 PMID:26670543

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

    PubMed Central

    Fang, Jie; Palanisami, Akilan; Rajapakshe, Kimal; Widger, William R.; Miller, John H.

    2011-01-01

    Nonlinear dielectric spectroscopy (NDS) is a non-invasive probe of cellular metabolic activity with potential application in the development of whole-cell biosensors. However, the mechanism of NDS interaction with metabolic membrane proteins is poorly understood, partly due to the inherent complexity of single cell organisms. Here we use the light-activated electron transport chain of spinach thylakoid membrane as a model system to study how NDS interacts with metabolic activity. We find protein modification, as opposed to membrane pump activity, to be the dominant source of NDS signal change in this system. Potential mechanisms for such protein modifications include reactive oxygen species generation and light-activated phosphorylation. PMID:25586698

  17. Membrane processes for removal of pharmaceutically active compounds (PhACs) from water and wastewaters.

    PubMed

    Taheran, Mehrdad; Brar, Satinder K; Verma, M; Surampalli, R Y; Zhang, T C; Valero, J R

    2016-03-15

    Pharmaceutically active compounds (PhACs), which find their way easily into the water sources, are emerging as a major concern for drinking water quality and aquatic species. Therefore, their removal from water sources is a priority from environmental point of view. During the past decade, different methods including membrane separation, adsorption systems and chemical transformation have been evaluated for removal of these compounds. This paper reviews different aspects of PhAC removal by using membrane separation processes, as they have been conventionally known to show high potential in the production of superior quality drinking and industrial water. In brief, osmosis membranes can efficiently remove almost all PhACs though its operational cost is relatively high and nanofiltration (NF) membranes are highly influenced by electrostatic and hydrophobic interaction. Moreover, the efficiency of membrane bioreactors (MBRs) is difficult to predict due to the complex interaction of compounds with microorganisms. To improve the performance and robustness of membrane technology, it is suggested to combine membranes with other systems, such as activated carbon and enzymatic degradation. PMID:26789358

  18. Controlled release and antibacterial activity of tetracycline hydrochloride-loaded bacterial cellulose composite membranes.

    PubMed

    Shao, Wei; Liu, Hui; Wang, Shuxia; Wu, Jimin; Huang, Min; Min, Huihua; Liu, Xiufeng

    2016-07-10

    Bacterial cellulose (BC) is widely used in biomedical applications. In this study, we prepared an antibiotic drug tetracycline hydrochloride (TCH)-loaded bacterial cellulose (BC) composite membranes, and evaluated the drug release, antibacterial activity and biocompatibility. The structure and morphology of the fabricated BC-TCH composite membranes were characterized using scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR). The TCH release results show that the incorporation of BC matrix to load TCH is able to control the release. In vitro antibacterial assay demonstrate that the developed BC-TCH composites displayed excellent antibacterial activity solely associated with the loaded TCH drug. More importantly, the BC-TCH composite membranes display good biocompatibility. These characteristics of BC-TCH composite membranes indicate that they may successfully serve as wound dressings and other medical biomaterials. PMID:27106158

  19. Luminol electrochemiluminescence for the analysis of active cholesterol at the plasma membrane in single mammalian cells.

    PubMed

    Ma, Guangzhong; Zhou, Junyu; Tian, Chunxiu; Jiang, Dechen; Fang, Danjun; Chen, Hongyuan

    2013-04-16

    A luminol electrochemiluminescence assay was reported to analyze active cholesterol at the plasma membrane in single mammalian cells. The cellular membrane cholesterol was activated by the exposure of the cells to low ionic strength buffer or the inhibition of intracellular acyl-coA/cholesterol acyltransferase (ACAT). The active membrane cholesterol was reacted with cholesterol oxidase in the solution to generate a peak concentration of hydrogen peroxide on the electrode surface, which induced a measurable luminol electrochemiluminescence. Further treatment of the active cells with mevastatin decreased the active membrane cholesterol resulting in a drop in luminance. No change in the intracellular calcium was observed in the presence of luminol and voltage, which indicated that our analysis process might not interrupt the intracellular cholesterol trafficking. Single cell analysis was performed by placing a pinhole below the electrode so that only one cell was exposed to the photomultiplier tube (PMT). Twelve single cells were analyzed individually, and a large deviation on luminance ratio observed exhibited the cell heterogeneity on the active membrane cholesterol. The smaller deviation on ACAT/HMGCoA inhibited cells than ACAT inhibited cells suggested different inhibition efficiency for sandoz 58035 and mevastatin. The new information obtained from single cell analysis might provide a new insight on the study of intracellular cholesterol trafficking. PMID:23527944

  20. Fast serial analysis of active cholesterol at the plasma membrane in single cells.

    PubMed

    Tian, Chunxiu; Zhou, Junyu; Wu, Zeng-Qiang; Fang, Danjun; Jiang, Dechen

    2014-01-01

    Previously, our group has utilized the luminol electrochemiluminescence to analyze the active cholesterol at the plasma membrane in single cells by the exposure of one cell to a photomultiplier tube (PMT) through a pinhole. In this paper, fast analysis of active cholesterol at the plasma membrane in single cells was achieved by a multimicroelectrode array without the pinhole. Single cells were directly located on the microelectrodes using cell-sized microwell traps. A cycle of voltage was applied on the microelectrodes sequentially to induce a peak of luminescence from each microelectrode for the serial measurement of active membrane cholesterol. A minimal time of 1.60 s was determined for the analysis of one cell. The simulation and the experimental data exhibited a semisteady-state distribution of hydrogen peroxide on the microelectrode after the reaction of cholesterol oxidase with the membrane cholesterol, which supported the relative accuracy of the serial analysis. An eight-microelectrode array was demonstrated to analyze eight single cells in 22 s serially, including the channel switching time. The results from 64 single cells either activated by low ion strength buffer or the inhibition of intracellular acyl-coA/cholesterol acyltransferase (ACAT) revealed that most of the cells analyzed had the similar active membrane cholesterol, while few cells had more active cholesterol resulting in the cellular heterogeneity. The fast single-cell analysis platform developed will be potentially useful for the analysis of more molecules in single cells using proper oxidases. PMID:24328095

  1. Specificity and mechanism of action of alpha-helical membrane-active peptides interacting with model and biological membranes by single-molecule force spectroscopy

    PubMed Central

    Sun, Shiyu; Zhao, Guangxu; Huang, Yibing; Cai, Mingjun; Shan, Yuping; Wang, Hongda; Chen, Yuxin

    2016-01-01

    In this study, to systematically investigate the targeting specificity of membrane-active peptides on different types of cell membranes, we evaluated the effects of peptides on different large unilamellar vesicles mimicking prokaryotic, normal eukaryotic, and cancer cell membranes by single-molecule force spectroscopy and spectrum technology. We revealed that cationic membrane-active peptides can exclusively target negatively charged prokaryotic and cancer cell model membranes rather than normal eukaryotic cell model membranes. Using Acholeplasma laidlawii, 3T3-L1, and HeLa cells to represent prokaryotic cells, normal eukaryotic cells, and cancer cells in atomic force microscopy experiments, respectively, we further studied that the single-molecule targeting interaction between peptides and biological membranes. Antimicrobial and anticancer activities of peptides exhibited strong correlations with the interaction probability determined by single-molecule force spectroscopy, which illustrates strong correlations of peptide biological activities and peptide hydrophobicity and charge. Peptide specificity significantly depends on the lipid compositions of different cell membranes, which validates the de novo design of peptide therapeutics against bacteria and cancers. PMID:27363513

  2. Specificity and mechanism of action of alpha-helical membrane-active peptides interacting with model and biological membranes by single-molecule force spectroscopy.

    PubMed

    Sun, Shiyu; Zhao, Guangxu; Huang, Yibing; Cai, Mingjun; Shan, Yuping; Wang, Hongda; Chen, Yuxin

    2016-01-01

    In this study, to systematically investigate the targeting specificity of membrane-active peptides on different types of cell membranes, we evaluated the effects of peptides on different large unilamellar vesicles mimicking prokaryotic, normal eukaryotic, and cancer cell membranes by single-molecule force spectroscopy and spectrum technology. We revealed that cationic membrane-active peptides can exclusively target negatively charged prokaryotic and cancer cell model membranes rather than normal eukaryotic cell model membranes. Using Acholeplasma laidlawii, 3T3-L1, and HeLa cells to represent prokaryotic cells, normal eukaryotic cells, and cancer cells in atomic force microscopy experiments, respectively, we further studied that the single-molecule targeting interaction between peptides and biological membranes. Antimicrobial and anticancer activities of peptides exhibited strong correlations with the interaction probability determined by single-molecule force spectroscopy, which illustrates strong correlations of peptide biological activities and peptide hydrophobicity and charge. Peptide specificity significantly depends on the lipid compositions of different cell membranes, which validates the de novo design of peptide therapeutics against bacteria and cancers. PMID:27363513

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

    PubMed

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

    2015-09-15

    Polysulfone membranes (PSf) containing silver nanoparticles were prepared by the wet phase-inversion process. Silver nanoparticles (AgNP) were dispersed into the polymer matrix using two different methodologies. In the first one, the AgNP were synthesized and further dispersed into the polymer solution (ex situ process). In the second method, the formation of the AgNP was performed in situ. The AgNP crystalline structure in the PSf membranes was confirmed by X-ray diffraction. Field emission scanning electron microscopy images showed that the addition of AgNP in PSf membranes caused no significant changes to the finger-like morphology. When the ex situ methodology was applied, 45 nm average size AgNP were uniformly distributed in the internal pores of the membranes. However, when the AgNP were formed through the in situ process, the AgNP were uniformly and preferentially distributed on the top and bottom surfaces of the membrane. In the last case, the AgNP showed cubic morphology when present in the bottom and top surfaces, however, when inside the membrane their morphology was spherical. The cubic-like nanoparticles displayed a 38 nm average edge length. The silver ion released from the membrane during water filtration was measured using inductively coupled plasma mass spectrometry, which showed a silver leaching of approximately 2 μg L(-1). The nanocomposite membranes prepared by the in situ method exhibited a better antibacterial activity, in comparison to those prepared by ex situ, and also a decrease in 90% Escherichia coli adhered cells compared to the pristine PSf membranes. In conclusion, the in situ procedure can be considered a feasible, simple, and reproducible methodology to prepare anti-biofouling polysulfone membranes containing AgNP. PMID:26099831

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

    PubMed Central

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

    2016-01-01

    Membrane active peptides are of large interest for development of drug delivery vehicles and therapeutics for treatment of multiple drug resistant infections. Lack of specificity can be detrimental and finding routes to tune specificity and activity of membrane active peptides is vital for improving their therapeutic efficacy and minimize harmful side effects. We describe a de novo designed membrane active peptide that partition into lipid membranes only when specifically and covalently anchored to the membrane, resulting in pore-formation. Dimerization with a complementary peptide efficiently inhibits formation of pores. The effect can be regulated by proteolytic digestion of the inhibitory peptide by the matrix metalloproteinase MMP-7, an enzyme upregulated in many malignant tumors. This system thus provides a precise and specific route for tuning the permeability of lipid membranes and a novel strategy for development of recognition based membrane active peptides and indirect enzymatically controlled release of liposomal cargo. PMID:26892926

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

    PubMed

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

    2016-01-01

    Membrane active peptides are of large interest for development of drug delivery vehicles and therapeutics for treatment of multiple drug resistant infections. Lack of specificity can be detrimental and finding routes to tune specificity and activity of membrane active peptides is vital for improving their therapeutic efficacy and minimize harmful side effects. We describe a de novo designed membrane active peptide that partition into lipid membranes only when specifically and covalently anchored to the membrane, resulting in pore-formation. Dimerization with a complementary peptide efficiently inhibits formation of pores. The effect can be regulated by proteolytic digestion of the inhibitory peptide by the matrix metalloproteinase MMP-7, an enzyme upregulated in many malignant tumors. This system thus provides a precise and specific route for tuning the permeability of lipid membranes and a novel strategy for development of recognition based membrane active peptides and indirect enzymatically controlled release of liposomal cargo. PMID:26892926

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

    NASA Astrophysics Data System (ADS)

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

    2016-02-01

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

  7. Biologically Inspired Photocatalytically Active Membranes for Water Treatment

    NASA Astrophysics Data System (ADS)

    Kinsinger, Nichola M.

    There is an alarming increase of a variety of new chemicals that are now being discharged into the wastewater system causing increased concern for public health and safety because many are not removed by typical wastewater treatment practices. Titanium Dioxide (TiO2) is a heterogeneous photocatalytic material that rapidly and completely mineralizing organics without harmful byproducts. TiO2 is synthesized by various methods, which lack the necessary control of crystal size, phase, and morphological features that yield optimized semiconductor materials. Mineralizing organisms demonstrate how nature can produce elegant structures at room temperature through controlled organic-mineral interactions. Here, we utilize biologically-inspired scaffolds to template the nucleation and growth of inorganic materials such as TiO2, which aid in controlling the size and phase of these particles and ultimately, their properties. Nanosized rutile and anatase particles were synthesized under solution conditions at relatively low temperatures and mild pH conditions. The effects of reaction conditions on phase and grain size were investigated and discussed from coordination chemistry and coarsening mechanisms. Photocatalytic characterization of TiO2 phase mixtures was performed to investigate their synergistic effect. The suspension conditions of these catalytic nanomaterials were modulated to optimize the degradation rate of organic analytes. Through the addition of an organic scaffold during the synthesis reaction, a mechanically robust (elastic) composite material containing TiO2 nanoparticles was produced. This composite was subsequently heat-treated to produce a porous, high surface area TiO2 nanoparticulate membrane. Processing conditions were investigated to characterize the growth and phase transformation of TiO2, which ultimately impacts photocatalytic performance. These bulk porous TiO2 structures can be fabricated and tailored to act as stand-alone photocatalytic membranes

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

    PubMed Central

    Cyboran, Sylwia; Żyłka, Romuald; Oszmiański, Jan; Kleszczyńska, Halina

    2014-01-01

    Compounds contained in fruits and leaves of blackcurrant (Ribes nigrum L.) are known as agents acting preventively and therapeutically on the organism. The HPLC analysis showed they are rich in polyphenol anthocyanins in fruits and flavonoids in leaves, that have antioxidant activity and are beneficial for health. The aim of the research was to determine the effect of blackcurrant fruit and leaf extracts on the physical properties of the erythrocyte membranes and assess their antioxidant properties. The effect of the extracts on osmotic resistance, shape of erythrocytes and hemolytic and antioxidant activity of the extracts were examined with spectrophotometric methods. The FTIR investigation showed that extracts modify the erythrocyte membrane and protect it against free radicals induced by UV radiation. The results show that the extracts do not induce hemolysis and even protect erythrocytes against the harmful action of UVC radiation, while slightly strengthening the membrane and inducing echinocytes. The compounds contained in the extracts do not penetrate into the hydrophobic region, but bind to the membrane surface inducing small changes in the packing arrangement of the polar head groups of membrane lipids. The extracts have a high antioxidant activity. Their presence on the surface of the erythrocyte membrane entails protection against free radicals. PMID:24527456

  9. Sphingomyelinase D Activity in Model Membranes: Structural Effects of in situ Generation of Ceramide-1-Phosphate

    PubMed Central

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

    2012-01-01

    The toxicity of Loxosceles spider venom has been attributed to a rare enzyme, sphingomyelinase D, which transforms sphingomyelin to ceramide-1-phosphate. The bases of its inflammatory and dermonecrotic activity, however, remain unclear. In this work the effects of ceramide-1-phosphate on model membranes were studied both by in situ generation of this lipid using a recombinant sphingomyelinase D from the spider Loxosceles laeta and by pre-mixing it with sphingomyelin and cholesterol. The systems of choice were large unilamellar vesicles for bulk studies (enzyme kinetics, fluorescence spectroscopy and dynamic light scattering) and giant unilamellar vesicles for fluorescence microscopy examination using a variety of fluorescent probes. The influence of membrane lateral structure on the kinetics of enzyme activity and the consequences of enzyme activity on the structure of target membranes containing sphingomyelin were examined. The findings indicate that: 1) ceramide-1-phosphate (particularly lauroyl ceramide-1-phosphate) can be incorporated into sphingomyelin bilayers in a concentration-dependent manner and generates coexistence of liquid disordered/solid ordered domains, 2) the activity of sphingomyelinase D is clearly influenced by the supramolecular organization of its substrate in membranes and, 3) in situ ceramide-1-phosphate generation by enzymatic activity profoundly alters the lateral structure and morphology of the target membranes. PMID:22558302

  10. Aryl-alkyl-lysines: Membrane-Active Small Molecules Active against Murine Model of Burn Infection.

    PubMed

    Ghosh, Chandradhish; Manjunath, Goutham B; Konai, Mohini M; Uppu, Divakara S S M; Paramanandham, Krishnamoorthy; Shome, Bibek R; Ravikumar, Raju; Haldar, Jayanta

    2016-02-12

    Infections caused by drug-resistant Gram-negative pathogens continue to be significant contributors to human morbidity. The recent advent of New Delhi metallo-β-lactamase-1 (blaNDM-1) producing pathogens, against which few drugs remain active, has aggravated the problem even further. This paper shows that aryl-alkyl-lysines, membrane-active small molecules, are effective in treating infections caused by Gram-negative pathogens. One of the compounds of the study was effective in killing planktonic cells as well as dispersing biofilms of Gram-negative pathogens. The compound was extremely effective in disrupting preformed biofilms and did not select resistant bacteria in multiple passages. The compound retained activity in different physiological conditions and did not induce any toxic effect in female Balb/c mice until concentrations of 17.5 mg/kg. In a murine model of Acinetobacter baumannii burn infection, the compound was able to bring the bacterial burden down significantly upon topical application for 7 days. PMID:27624962

  11. Multiple sources of carbonic anhydrase activity in pea thylakoids: soluble and membrane-bound forms.

    PubMed

    Rudenko, Natalia N; Ignatova, Lyudmila K; Ivanov, Boris N

    2007-01-01

    Carbonic anhydrase (CA) activity of pea thylakoids, thylakoid membranes enriched with photosystem I (PSI-membranes), or photosystem II (PSII-membranes) as well as both supernatant and pellet after precipitation of thylakoids treated with detergent Triton X-100 were studied. CA activity of thylakoids in the presence of varying concentrations of Triton X-100 had two maxima, at Triton/chlorophyll (triton/Chl) ratios of 0.3 and 1.0. CA activities of PSI-membranes and PSII-membranes had only one maximum each, at Triton/Chl ratio 0.3 or 1.0, respectively. Two CAs with characteristics of the membrane-bound proteins and one CA with characteristics of the soluble proteins were found in the medium after thylakoids were incubated with Triton. One of the first two CAs had mobility in PAAG after native electrophoresis the same as that of CA residing in PSI-membranes, and the other CA had mobility the same as the mobility of CA residing in PSII-membranes, but the latter was different from CA situated in PSII core-complex (Ignatova et al. 2006 Biochemistry (Moscow) 71:525-532). The properties of the "soluble" CA removed from thylakoids were different from the properties of the known soluble CAs of plant cell: apparent molecular mass was about 262 kD and it was three orders more sensitive to the specific CA inhibitor, ethoxyzolamide, than soluble stromal CA. The data are discussed as indicating the presence of, at least, four CAs in pea thylakoids. PMID:17347907

  12. Cognition Is Cool: Can Hemispheric Activation Be Assessed by Tympanic Membrane Thermometry?

    ERIC Educational Resources Information Center

    Cherbuin, Nicolas; Brinkman, Cobie

    2004-01-01

    Hemispheric activation during cognitive tasks using functional magnetic resonance imaging (fMRI) can be difficult to interpret, uncomfortable, and is not widely available. This study investigated whether tympanic membrane thermometry could be used as a broad measure of hemispheric activation. Infrared probes measured ear temperature continuously…

  13. [Effect of powdered activated carbon on the sludge mixed liquor characteristics and membrane fouling of MBR].

    PubMed

    Li, Shao-Feng; Gao, Yuan

    2011-02-01

    Effect of dosing powder activated carbon (PAC) on the characteristics of the sludge mixed liquor in membrane bioreactor (MBR) was investigated by parallel tests. And the reason that PAC mitigated membrane fouling was also explored. The results showed that PAC could decrease mixture viscosity and increase sludge particle size, which led to less trans-membrane pressure developing. Extracellular polymer substances (EPS) content, sludge specific resistance and cake layer resistance (R(c)) had a good correlation. Adding PAC could decrease EPS concentration, sludge specific resistance and then slow down the increase of R(c), which mitigated membrane fouling. Membrane pore blocking resistance (R(p)) increased exponentially with increasing of the soluble microbial products (SMP) concentration in the supernatant. Dosing PAC reduced the SMP concentration and slowed down the growth rate of R(p), which was helpful to mitigating membrane fouling. R(c) and R(p) increased along with the operation of MBRs and R(c)/R(f) (26.32% -63.16%) was always greater than R(p)/R(f) (7.89% -35.32%) which suggested the R(c) was the main factor in membrane fouling. Moreover, it was also found that controlling of dosing PAC on R(c) was better than it on R(p). PMID:21528575

  14. Comparing graphene, carbon nanotubes, and superfine powdered activated carbon as adsorptive coating materials for microfiltration membranes.

    PubMed

    Ellerie, Jaclyn R; Apul, Onur G; Karanfil, Tanju; Ladner, David A

    2013-10-15

    Multi-walled carbon nanotubes (MWCNTs), nano-graphene platelets (NGPs), and superfine powdered activated carbon (S-PAC) were comparatively evaluated for their applicability as adsorptive coatings on microfiltration membranes. The objective was to determine which materials were capable of contaminant removal while causing minimal flux reduction. Methylene blue and atrazine were the model contaminants. When applied as membrane coatings, MWCNTs had minimal retention capabilities for the model contaminants, and S-PAC had the fastest removal. The membrane coating approach was also compared with a stirred vessel configuration, in which the adsorbent was added to a stirred flask preceding the membrane cell. Direct application of the adsorbent to the membrane constituted a greater initial reduction in permeate concentrations of the model contaminants than with the stirred flask setup. All adsorbents except S-PAC showed flux reductions less than 5% after application as thin-layer membrane coatings, and flux recovery after membrane backwashing was greater than 90% for all materials and masses tested. PMID:23911830

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

    NASA Astrophysics Data System (ADS)

    Pujiastuti, Sri; Onggo, Holia

    2016-02-01

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

  16. Nonbonded interactions in membrane active cyclic biopolymers. IV - Cation dependence

    NASA Technical Reports Server (NTRS)

    Radhakrishnan, R.; Srinivasan, S.; Prasad, C. V.; Brinda, S. R.; Macelroy, R. D.; Sundaram, K.

    1980-01-01

    Interactions of valinomycin and form of its analogs in several conformations with the central ions Li(+), Na(+), K(+), Rb(+) and Cs(+) are investigated as part of a study of the specific preference of valinomycin for potassium and the mechanisms of carrier-mediated ion transport across membranes. Ion binding energies and conformational potential energies are calculated taking into account polarization energy formulas and repulsive energy between the central ion and the ligand atoms for conformations representing various stages in ion capture and release for each of the two ring chiralities of valinomycin and its analogs. Results allow the prediction of the chirality and conformation most likely to be observed for a given analog, and may be used to synthesize analogs with a desired rigidity or flexibility. The binding energies with the alkali metal cations are found to decrease with increasing ion size, and to be smaller than the corresponding ion hydration energies. It is pointed out that the observed potassium preference may be explainable in terms of differences between binding and hydration energies. Binding energies are also noted to depend on ligand conformation.

  17. Protein kinase and phosphatase activities of thylakoid membranes

    SciTech Connect

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

    1987-01-01

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

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

    SciTech Connect

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

    1986-05-01

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

  19. Instant membrane resealing in nlrp3 inflammmasome activation of endothelial cells.

    PubMed

    Chen, Yang; Yuan, Ming; Xia, Min; Wang, Lei; Zhang, Yang; Li, Pin-Lan

    2016-01-01

    The present study explored the molecular mechanisms by which instant cell membrane resealing (CMR) controls the activation of NOD-like receptor pyrin domain containing 3 (Nlrp3) inflammasomes. Using wavelength-switching fluorescent microscopy with PI and fura-2 as indicators, we monitored instant CMR simultaneously with (Ca(2+))i in mouse microvascular endothelial cell (MVECs). LCWE or saponin wad found to produce membrane injury, which was resealed in a Ca(2+)-dependent manner, but abolished by FasL, a membrane raft (MR) clustering stimulator. Even in the presence of Ca(2+), FasL prolonged the CMR time as shown by an earlier onset of PI influx (48±12 sec vs. 17±3 min. of control). These effects of FasL were substantially blocked by an MR disruptor, methyl-beta-cyclodextrin (MCD). The failure of CMR upon FasL activated Nlrp3 inflammasomes, which was blocked by MCD, a membrane resealing compound, VA64 or siRNA of an MR-facilitating enzyme, acid sphingomyelinase. This inflammasome activation was due to increased lysosomal permeability and cathepsin B release. It is concluded that an MR-associated CMR protects ECs from Nlrp3 inflammasome activation induced by membrane injury. PMID:26709796

  20. Requirement for membrane potential in active transport of glutamine by Escherichia coli.

    PubMed Central

    Plate, C A

    1979-01-01

    The effect of reducing the membrane potential on glutamine transport in cells of Escherichia coli has been investigated. Addition of valinomycin to tris(hydroxymethyl)aminomethane-ethylenediaminetetraacetic acid-treated E. coli cells in the presence of 20 mM exogenous potassium reduced the membrane potential, as measured by the uptake of the lipophilic cation triphenylmethylphosphonium, and caused a complete inhibition of glutamine transport. Valinomycin plus potassium also caused a rapid decrease in the intracellular levels of ATP of normal E. coli cells, but had little if any effect on the ATP levels of two mutants of E. coli carrying lesions in the energy-transducing ATP complex (unc mutants). Yet both the membrane potential and the capacity to transport glutamine were depressed in the unc mutants by valinomycin and potassium. These findings are consistent with the hypothesis that both ATP and a membrane potential are essential to the active transport of glutamine by E. coli cells. PMID:153897

  1. [Effect of alkylresorcin on biological membranes during activation of lipid peroxidation].

    PubMed

    Erin, A N; Davitashvili, N G; Prilipko, L L; Boldyrev, A A; Lushchak, V I

    1987-07-01

    The effect of alkyl resorcin isolated from the cells of Azotobacter chroococcum and of its structural analog devoid of the alkyl chain (resorcin) on liver microsomes and brain synaptosomes of the rat as well as on rabbit skeletal muscle sarcoplasmic reticulum fragments during activation of lipid peroxidation was studied. Alkyl resorcin was shown to produce a much more potent antioxidant effect as compared with resorcin, since it inhibited lipid peroxidation in all the three types of membranes under study at much lower concentrations. Both alkyl resorcin and resorcin which inhibit lipid peroxidation prevented lipid peroxidation-induced structural-functional damages of synaptosomal and sarcoplasmic reticulum fragment membranes. Unlike resorcin, alkyl resorcin exerted an additional effect on brain synaptosomal membranes which consisted in the stabilization of barrier functions of membranes during incomplete inhibition of lipid peroxidation. The cumulative data suggest that stabilization necessitates the presence of both resorcin radical and alkyl chain in the alkyl resorcin molecule. PMID:3663757

  2. Importance of Membrane Structural Integrity for RPE65 Retinoid Isomerization Activity*

    PubMed Central

    Golczak, Marcin; Kiser, Philip D.; Lodowski, David T.; Maeda, Akiko; Palczewski, Krzysztof

    2010-01-01

    Regeneration of visual chromophore in the vertebrate visual cycle involves the retinal pigment epithelium-specific protein RPE65, the key enzyme catalyzing the cleavage and isomerization of all-trans-retinyl fatty acid esters to 11-cis-retinol. Although RPE65 has no predicted membrane spanning domains, this protein predominantly associates with microsomal fractions isolated from bovine retinal pigment epithelium (RPE). We have re-examined the nature of RPE65 interactions with native microsomal membranes by using extraction and phase separation experiments. We observe that hydrophobic interactions are the dominant forces that promote RPE65 association with these membranes. These results are consistent with the crystallographic model of RPE65, which features a large lipophilic surface that surrounds the entrance to the catalytic site of this enzyme and likely interacts with the hydrophobic core of the endoplasmic reticulum membrane. Moreover, we report a critical role for phospholipid membranes in preserving the retinoid isomerization activity and physical properties of RPE65. Isomerase activity measured in bovine RPE was highly sensitive to phospholipase A2 treatment, but the observed decline in 11-cis-retinol production did not directly reflect inhibition by products of lipid hydrolysis. Instead, a direct correlation between the kinetics of phospholipid hydrolysis and retinoid isomerization suggests that the lipid membrane structure is critical for RPE65 enzymatic activity. We also provide evidence that RPE65 operates in a multiprotein complex with retinol dehydrogenase 5 and retinal G protein-coupled receptor in RPE microsomes. Modifications in the phospholipid environment affecting interactions with these protein components may be responsible for the alterations in retinoid metabolism observed in phospholipid-depleted RPE microsomes. Thus, our results indicate that the enzymatic activity of native RPE65 strongly depends on its membrane binding and phospholipid

  3. Ultrathin and stable active layer of dense composite membrane enabled by poly(dopamine).

    PubMed

    Li, Ben; Liu, Wanpeng; Jiang, Zhongyi; Dong, Xiao; Wang, Baoyi; Zhong, Yurong

    2009-07-01

    We demonstrate that dopamine is able to self-polymerize and adhere firmly onto the substrate, which can create a hierarchical structure comprising an ultrathin active layer and a porous support layer. Such an approach opens a novel way to fabricating highly efficient and stable composite materials including composite membranes. More specifically, in this study the composite membranes are fabricated by simply dipping microporous substrate in aqueous dopamine solution under mild conditions. Nanoindentation measurement reveals the tight adhesion of dopamine onto microporous substrate, which is ascribed to numerous pi-pi and hydrogen-bonding interactions. The chemical composition of the active layer is analyzed by XPS, which demonstrates the self-polymerization of dopamine. The water contact angle of the dopamine coated membranes is reduced remarkably compared with that of the uncoated counterpart. Stylus profiler measurements display that the poly(dopamine) thickness increases as the coating time increases. FESEM images of the membranes' cross section show that an active layer (<100 nm) is deposited on the porous polysulfone (PS) substrate. Positron annihilation spectroscopy (PAS) is introduced to probe the fractional free volume properties throughout the cross section of the composite membranes and reveal that after dopamine double-coating the active layer becomes thicker and more compact. Moreover, pH and concentration of the dopamine solution exert notable influence on the fractional free volume of the composite membranes. The as-prepared membranes are tentatively employed for pervaporative desulfurization and exhibits satisfying separation performance as well as durability. This facile, versatile, and efficient approach enables a promising prospect for the wide applications of such novel kinds of ultrathin composite materials. PMID:19366196

  4. Na+/H+ Exchange Activity in the Plasma Membrane of Arabidopsis1

    PubMed Central

    Qiu, Quan-Sheng; Barkla, Bronwyn J.; Vera-Estrella, Rosario; Zhu, Jian-Kang; Schumaker, Karen S.

    2003-01-01

    In plants, Na+/H+ exchangers in the plasma membrane are critical for growth in high levels of salt, removing toxic Na+ from the cytoplasm by transport out of the cell. The molecular identity of a plasma membrane Na+/H+ exchanger in Arabidopsis (SOS1) has recently been determined. In this study, immunological analysis provided evidence that SOS1 localizes to the plasma membrane of leaves and roots. To characterize the transport activity of this protein, purified plasma membrane vesicles were isolated from leaves of Arabidopsis. Na+/H+ exchange activity, monitored as the ability of Na to dissipate an established pH gradient, was absent in plants grown without salt. However, exchange activity was induced when plants were grown in 250 mm NaCl and increased with prolonged salt exposure up to 8 d. H+-coupled exchange was specific for Na, because chloride salts of other monovalent cations did not dissipate the pH gradient. Na+/H+ exchange activity was dependent on Na (substrate) concentration, and kinetic analysis indicated that the affinity (apparent Km) of the transporter for Na+ is 22.8 mm. Data from two experimental approaches supports electroneutral exchange (one Na+ exchanged for one proton): (a) no change in membrane potential was measured during the exchange reaction, and (b) Na+/H+ exchange was unaffected by the presence or absence of a membrane potential. Results from this research provide a framework for future studies into the regulation of the plant plasma membrane Na+/H+ exchanger and its relative contribution to the maintenance of cellular Na+ homeostasis during plant growth in salt. PMID:12805632

  5. Importance of Membrane Structural Integrity for RPE65 Retinoid Isomerization Activity

    SciTech Connect

    Golczak, Marcin; Kiser, Philip D.; Lodowski, David T.; Maeda, Akiko; Palczewski, Krzysztof

    2010-04-05

    Regeneration of visual chromophore in the vertebrate visual cycle involves the retinal pigment epithelium-specific protein RPE65, the key enzyme catalyzing the cleavage and isomerization of all-trans-retinyl fatty acid esters to 11-cis-retinol. Although RPE65 has no predicted membrane spanning domains, this protein predominantly associates with microsomal fractions isolated from bovine retinal pigment epithelium (RPE). We have re-examined the nature of RPE65 interactions with native microsomal membranes by using extraction and phase separation experiments. We observe that hydrophobic interactions are the dominant forces that promote RPE65 association with these membranes. These results are consistent with the crystallographic model of RPE65, which features a large lipophilic surface that surrounds the entrance to the catalytic site of this enzyme and likely interacts with the hydrophobic core of the endoplasmic reticulum membrane. Moreover, we report a critical role for phospholipid membranes in preserving the retinoid isomerization activity and physical properties of RPE65. Isomerase activity measured in bovine RPE was highly sensitive to phospholipase A{sup 2} treatment, but the observed decline in 11-cis-retinol production did not directly reflect inhibition by products of lipid hydrolysis. Instead, a direct correlation between the kinetics of phospholipid hydrolysis and retinoid isomerization suggests that the lipid membrane structure is critical for RPE65 enzymatic activity. We also provide evidence that RPE65 operates in a multiprotein complex with retinol dehydrogenase 5 and retinal G protein-coupled receptor in RPE microsomes. Modifications in the phospholipid environment affecting interactions with these protein components may be responsible for the alterations in retinoid metabolism observed in phospholipid-depleted RPE microsomes. Thus, our results indicate that the enzymatic activity of native RPE65 strongly depends on its membrane binding and

  6. Membrane composition influences the activity of in vitro refolded human vitamin K epoxide reductase.

    PubMed

    Jaenecke, Frank; Friedrich-Epler, Beatrice; Parthier, Christoph; Stubbs, Milton T

    2015-10-27

    Human vitamin K epoxide reductase (hVKOR) is an integral membrane protein responsible for the maintenance of reduced vitamin K pools, a prerequisite for the action of γ-glutamyl carboxylase and hence for hemostasis. Here we describe the recombinant expression of hVKOR as an insoluble fusion protein in Escherichia coli, followed by purification and chemical cleavage under denaturing conditions. In vitro renaturation and reconstitution of purified solubilized hVKOR in phospholipids could be established to yield active protein. Crucially, the renatured enzyme is inhibited by the powerful coumarin anticoagulant warfarin, and we demonstrate that enzyme activity depends on lipid composition. The completely synthetic system for protein production allows a rational investigation of the multiple variables in membrane protein folding and paves the way for the provision of pure, active membrane protein for structural studies. PMID:26435421

  7. Sirtuin Activation: A Role for Plasma Membrane in the Cell Growth Puzzle

    PubMed Central

    2013-01-01

    For more than 20 years, the observation that impermeable oxidants can stimulate cell growth has not been satisfactorily explained. The discovery of sirtuins provides a logical answer to the puzzle. The NADH-dependent transplasma membrane electron transport system, which is stimulated by growth factors and interventions such as calorie restriction, can transfer electrons to external acceptors and protect against stress-induced apoptosis. We hypothesize that the activation of plasma membrane electron transport contributes to the cytosolic NAD+ pool required for sirtuin to activate transcription factors necessary for cell growth and survival. PMID:23033342

  8. delta-Opioid receptors exhibit high efficiency when activating trimeric G proteins in membrane domains.

    PubMed

    Bourova, Lenka; Kostrnova, Alexandra; Hejnova, Lucie; Moravcova, Zuzana; Moon, Hyo-Eun; Novotny, Jiri; Milligan, Graeme; Svoboda, Petr

    2003-04-01

    Low-density membrane fragments (domains) were separated from the bulk of plasma membranes of human embryonic kidney (HEK)293 cells expressing a delta-opioid (DOP) receptor-Gi1alpha fusion protein by drastic homogenization and flotation on equilibrium sucrose density gradients. The functional activity of trimeric G proteins and capacity of the DOP receptor to stimulate both the fusion protein-linked Gi1alpha and endogenous pertussis-toxin sensitive G proteins was measured as d-Ala2, d-Leu5-enkephalin stimulated high-affinity GTPase or guanosine-5'-[gamma-35S]triphosphate ([35S]GTPgammaS) binding. The maximum d-Ala2-d-Leu5 enkephalin (DADLE)-stimulated GTPase was two times higher in low-density membrane fragments than in bulk of plasma membranes; 58 and 27 pmol/mg/min, respectively. The same difference was obtained for [35S]GTPgammaS binding. Contrarily, the low-density domains contained no more than half the DOP receptor binding sites (Bmax = 6.6 pmol/mg versus 13.6 pmol/mg). Thus, when corrected for expression levels of the receptor, low-density domains exhibited four times higher agonist-stimulated GTPase and [35S]GTPgammaS binding than the bulk plasma membranes. The regulator of G protein signaling RGS1, enhanced further the G protein functional activity but did not remove the difference between domain-bound and plasma membrane pools of G protein. The potency of the agonist in functional studies and the affinity of specific [3H]DADLE binding to the receptor were, however, the same in both types of membranes - EC50 = 4.5 +/- 0.1 x 10(-8) and 3.2 +/- 1.4 x 10(-8) m for GTPase; Kd = 1.2 +/- 0.1 and 1.3 +/- 0.1 nm for [3H]DADLE radioligand binding assay. Similar results were obtained when sodium bicarbonate was used for alkaline isolation of membrane domains. By contrast, detergent-insensitive membrane domains isolated following treatment of cells with Triton X100 exhibited no DADLE-stimulated GTPase or GTPgammaS binding. Functional coupling between the DOP receptor

  9. Finding Large Aperture Fractures in Geothermal Resource Areas Using a Three-Component Long-Offset Surface Seismic Survey, PSInSAR and Kinematic Structural Analysis

    SciTech Connect

    Teplow, William J.; Warren, Ian

    2015-08-12

    The DOE cost-share program applied innovative and cutting edge seismic surveying and processing, permanent scatter interferometry-synthetic aperture radar (PSInSAR) and structural kinematics to the exploration problem of locating and mapping largeaperture fractures (LAFs) for the purpose of targeting geothermal production wells. The San Emidio geothermal resource area, which is under lease to USG, contains production wells that have encountered and currently produce from LAFs in the southern half of the resource area (Figure 2). The USG lease block, incorporating the northern extension of the San Emidio geothermal resource, extends 3 miles north of the operating wellfield. The northern lease block was known to contain shallow thermal waters but was previously unexplored by deep drilling. Results of the Phase 1 exploration program are described in detail in the Phase 1 Final Report (Teplow et al., 2011). The DOE cost shared program was completed as planned on September 30, 2014. This report summarizes results from all of Phase 1 and 2 activities.

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

    PubMed

    Uppu, Divakara S S M; Manjunath, Goutham B; Yarlagadda, Venkateswarlu; Kaviyil, Jyothi E; Ravikumar, Raju; Paramanandham, Krishnamoorthy; Shome, Bibek R; Haldar, Jayanta

    2015-01-01

    Gram-negative 'superbugs' such as New Delhi metallo-beta-lactamase-1 (blaNDM-1) producing pathogens have become world's major public health threats. Development of molecular strategies that can rehabilitate the 'old antibiotics' and halt the antibiotic resistance is a promising approach to target them. We report membrane-active macromolecules (MAMs) that restore the antibacterial efficacy (enhancement by >80-1250 fold) of tetracycline antibiotics towards blaNDM-1 Klebsiella pneumonia and blaNDM-1 Escherichia coli clinical isolates. Organismic studies showed that bacteria had an increased and faster uptake of tetracycline in the presence of MAMs which is attributed to the mechanism of re-sensitization. Moreover, bacteria did not develop resistance to MAMs and MAMs stalled the development of bacterial resistance to tetracycline. MAMs displayed membrane-active properties such as dissipation of membrane potential and membrane-permeabilization that enabled higher uptake of tetracycline in bacteria. In-vivo toxicity studies displayed good safety profiles and preliminary in-vivo antibacterial efficacy studies showed that mice treated with MAMs in combination with antibiotics had significantly decreased bacterial burden compared to the untreated mice. This report of re-instating the efficacy of the antibiotics towards blaNDM-1 pathogens using membrane-active molecules advocates their potential for synergistic co-delivery of antibiotics to combat Gram-negative superbugs. PMID:25789871

  11. Evidence of steroid hormone activity in the chorioallantoic membrane of a Turtle (Pseudemys nelsoni).

    PubMed

    Cruze, Lori; Hamlin, Heather J; Kohno, Satomi; McCoy, Michael W; Guillette, Louis J

    2013-06-01

    Endocrine properties of extraembryonic membranes have traditionally been viewed as a characteristic of placental amniotes. However, our laboratory recently demonstrated that this ability extends to the extraembryonic membranes of two oviparous amniotes (chicken and alligator) indicating that endocrine extraembryonic membranes are not an innovation of placental amniotes and suggesting that this could be a shared amniote characteristic. In this study, we test our hypothesis that the chorioallantoic membrane (CAM) obtained from non-archosaurian obligate oviparous amniotes such as turtles, have the potential for steroid hormone activity. To investigate synthesis of a major placental hormone, we performed explant culture and found that the turtle CAM synthesizes progesterone in vitro in the presence of a steroid precursor. In addition, to examine whether the CAM has the ability to respond to steroid signaling, we quantified mRNA expression of the progesterone, androgen, and two estrogen receptors. Finally, to determine if steroid receptor mRNA is translated to protein, we performed immunolocalization of the progesterone receptor. Our data demonstrate that the turtle CAM exhibits steroid synthesis and has steroid hormone signaling capabilities. To that end, steroid hormone activity has now been demonstrated in the CAMs of three oviparous species that represent three independent lineages within oviparous Reptilia that have never exhibited viviparity; thus these data support our hypothesis that endocrine activity of extraembryonic membranes is a conserved trait of Amniota. PMID:23458289

  12. Membrane-Active Macromolecules Resensitize NDM-1 Gram-Negative Clinical Isolates to Tetracycline Antibiotics

    PubMed Central

    Uppu, Divakara S. S. M.; Manjunath, Goutham B.; Yarlagadda, Venkateswarlu; Kaviyil, Jyothi E.; Ravikumar, Raju; Paramanandham, Krishnamoorthy; Shome, Bibek R.; Haldar, Jayanta

    2015-01-01

    Gram-negative ‘superbugs’ such as New Delhi metallo-beta-lactamase-1 (blaNDM-1) producing pathogens have become world’s major public health threats. Development of molecular strategies that can rehabilitate the ‘old antibiotics’ and halt the antibiotic resistance is a promising approach to target them. We report membrane-active macromolecules (MAMs) that restore the antibacterial efficacy (enhancement by >80-1250 fold) of tetracycline antibiotics towards blaNDM-1 Klebsiella pneumonia and blaNDM-1 Escherichia coli clinical isolates. Organismic studies showed that bacteria had an increased and faster uptake of tetracycline in the presence of MAMs which is attributed to the mechanism of re-sensitization. Moreover, bacteria did not develop resistance to MAMs and MAMs stalled the development of bacterial resistance to tetracycline. MAMs displayed membrane-active properties such as dissipation of membrane potential and membrane-permeabilization that enabled higher uptake of tetracycline in bacteria. In-vivo toxicity studies displayed good safety profiles and preliminary in-vivo antibacterial efficacy studies showed that mice treated with MAMs in combination with antibiotics had significantly decreased bacterial burden compared to the untreated mice. This report of re-instating the efficacy of the antibiotics towards blaNDM-1 pathogens using membrane-active molecules advocates their potential for synergistic co-delivery of antibiotics to combat Gram-negative superbugs. PMID:25789871

  13. Activation and routing of membrane-tethered prohormone convertases 1 and 2.

    PubMed

    Bruzzaniti, A; Marx, R; Mains, R E

    1999-08-27

    Many peptide hormones and neuropeptides are processed by members of the subtilisin-like family of prohormone convertases (PCs), which are either soluble or integral membrane proteins. PC1 and PC2 are soluble PCs that are primarily localized to large dense core vesicles in neurons and endocrine cells. We examined whether PC1 and PC2 were active when expressed as membrane-tethered proteins, and how tethering to membranes alters the biosynthesis, enzymatic activity, and intracellular routing of these PCs. PC1 and PC2 chimeras were constructed using the transmembrane domain and cytoplasmic domain of the amidating enzyme, peptidylglycine alpha-amidating monooxygenase (PAM). The membrane-tethered PCs were rerouted from large dense core vesicles to the Golgi region. In addition, the chimeras were transiently expressed at the cell surface and rapidly internalized to the Golgi region in a fashion similar to PAM. Membrane-tethered PC1 and PC2 exhibited changes in pro-domain maturation rates, N-glycosylation, and in the pH and calcium optima required for maximal enzymatic activity against a fluorogenic substrate. In addition, the PC chimeras efficiently cleaved endogenous pro-opiomelanocortin to the correct bioactive peptides. The PAM transmembrane domain/cytoplasmic domain also prevented stimulated secretion of pro-opiomelanocortin products in AtT-20 cells. PMID:10455138

  14. Effects of Gangliosides on the Activity of the Plasma Membrane Ca2+-ATPase

    PubMed Central

    Jiang, Lei; Bechtel, Misty D.; Bean, Jennifer L.; Winefield, Robert; Williams, Todd D.; Zaidi, Asma; Michaelis, Elias K.; Michaelis, Mary L.

    2014-01-01

    Control of intracellular calcium concentrations ([Ca2+]i) is essential for neuronal function, and the plasma membrane Ca2+-ATPase (PMCA) is crucial for the maintenance of low [Ca2+]i. We previously reported on loss of PMCA activity in brain synaptic membranes during aging. Gangliosides are known to modulate Ca2+ homeostasis and signal transduction in neurons. In the present study, we observed age-related changes in the ganglioside composition of synaptic plasma membranes. This led us to hypothesize that alterations in ganglioside species might contribute to the age-associated loss of PMCA activity. To probe the relationship between changes in endogenous ganglioside content or composition and PMCA activity in membranes of cortical neurons, we induced depletion of gangliosides by treating neurons with D-threo-1-phenyl-2-decanoylamino-3-morpholino-1-propanol (D-PDMP). This caused a marked decrease in the activity of PMCA, which suggested a direct correlation between ganglioside content and PMCA activity. Neurons treated with neuraminidase exhibited an increase in GM1 content, a loss in poly-sialoganglioside content, and a decrease in PMCA activity that was greater than that produced by D-PDMP treatment. Thus, it appeared that poly-sialogangliosides had a stimulatory effect whereas mono-sialogangliosides had the opposite effect. Our observations add support to previous reports of PMCA regulation by gangliosides by demonstrating that manipulations of endogenous ganglioside content and species affect the activity of PMCA in neuronal membranes. Furthermore, our studies suggest that age-associated loss in PMCA activity may result in part from changes in the lipid environment of this Ca2+ transporter. PMID:24434060

  15. Ionophore-Based Voltammetric Ion Activity Sensing with Thin Layer Membranes.

    PubMed

    Cuartero, Maria; Crespo, Gaston A; Bakker, Eric

    2016-02-01

    As shown in recent work, thin layer ion-selective multi-ionophore membranes can be interrogated by cyclic voltammetry to detect the ion activity of multiple species simultaneously and selectively. Additional fundamental evidence is put forward on ion discrimination with thin multi-ionophore-based membranes with thicknesses of 200 ± 25 nm and backside contacted with poly-3-octylthiophene (POT). An anodic potential scan partially oxidizes the POT film (to POT(+)), thereby initiating the release of hydrophilic cations from the membrane phase to the sample solution at a characteristic potential. Varying concentration of added cation-exchanger demonstrates that it limits the ion transfer charge and not the deposited POT film. Voltammograms with multiple peaks are observed with each associated with the transfer of one type of ion (lithium, potassium, and sodium). Experimental conditions (thickness and composition of the membrane and concentration of the sample) are chosen that allow one to describe the system by a thermodynamic rather than kinetic model. As a consequence, apparent stability constants for sodium, potassium, and lithium (assuming 1:1 stoichiometry) with their respective ionophores are calculated and agree well with the values obtained by the potentiometric sandwich membrane technique. As an analytical application, a membrane containing three ionophores was used to determine lithium, sodium, and potassium in artificial samples at the same location and within a single voltammetric scan. Lithium and potassium were also determined in undiluted human plasma in the therapeutic concentration range. PMID:26712342

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

    PubMed

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

    2016-05-18

    In this study, the bambara protein isolate (BPI) was digested with three proteases (alcalase, trypsin and pepsin), to produce bambara protein hydrolysates (BPHs). These hydrolysates were passed through ultrafiltration membranes to obtain peptide fractions of different sizes (<1, 1-3, 3-5 and 5-10 kDa). The hydrolysates and their peptide fractions were investigated for antioxidant activities. The membrane fractions showed that peptides with sizes <3 kDa had significantly (p < 0.05) reduced surface hydrophobicity when compared with peptides >3 kDa. This is in agreement with the result obtained for the ferric reducing power, metal chelating and hydroxyl radical scavenging activities where higher molecular weight peptides exhibited better activity (p < 0.05) when compared to low molecular weight peptide fractions. However, for all the hydrolysates, the low molecular weight peptides were more effective diphenyl-1-picrylhydrazyl (DPPH) radical scavengers but not superoxide radicals when compared to the bigger peptides. In comparison with glutathione (GSH), BPHs and their membrane fractions had better (p < 0.05) reducing power and ability to chelate metal ions except for the pepsin hydrolysate and its membrane fractions that did not show any metal chelating activity. However, the 5-10 kDa pepsin hydrolysate peptide fractions had greater (88%) hydroxyl scavenging activity than GSH, alcalase and trypsin hydrolysates (82%). These findings show the potential use of BPHs and their peptide fraction as antioxidants in reducing food spoilage or management of oxidative stress-related metabolic disorders. PMID:27156453

  17. Modulation of plasma membrane H+-ATPase activity differentially activates wound and pathogen defense responses in tomato plants.

    PubMed Central

    Schaller, A; Oecking, C

    1999-01-01

    Systemin is an important mediator of wound-induced defense gene activation in tomato plants, and it elicits a rapid alkalinization of the growth medium of cultured Lycopersicon peruvianum cells. A possible mechanistic link between proton fluxes across the plasma membrane and the induction of defense genes was investigated by modulating plasma membrane H+-ATPase activity. Inhibitors of H+-ATPase (erythrosin B, diethyl stilbestrol, and vanadate) were found to alkalinize the growth medium of L. peruvianum cell cultures and to induce wound response genes in whole tomato plants. Conversely, an activator of the H+-ATPase (fusicoccin) acidified the growth medium of L. peruvianum cell cultures and suppressed systemin-induced medium alkalinization. Likewise, in fusicoccin-treated tomato plants, the wound- and systemin-triggered accumulation of wound-responsive mRNAs was found to be suppressed. However, fusicoccin treatment of tomato plants led to the accumulation of salicylic acid and the expression of pathogenesis-related genes. Apparently, the wound and pathogen defense signaling pathways are differentially regulated by changes in the proton electrochemical gradient across the plasma membrane. In addition, alkalinization of the L. peruvianum cell culture medium was found to depend on the influx of Ca2+ and the activity of a protein kinase. Reversible protein phosphorylation was also shown to be involved in the induction of wound response genes. The plasma membrane H+-ATPase as a possible target of a Ca2+-activated protein kinase and its role in defense signaling are discussed. PMID:9927643

  18. Enzymatic activity of soluble and membrane tethered peptide pro-hormone convertase 1.

    PubMed

    Bruzzaniti, Angela; Mains, Richard E

    2002-05-01

    Pro-hormone convertases PC1 and PC2 perform endoproteolytic cleavages of precursors in peptide-containing secretory granules. PC1 and PC2 are soluble, secreted with bioactive peptides. Evolutionarily related PCs have membrane tethers, not secreted. We tethered PC1 to the transmembrane-cytoplasmic domains (CD) of a granule enzyme (peptidylglycine-alpha-amidating monooxygenase; PAM) and Golgi-localized PC8. The tethered PC1 is far more stable to elevated temperature and denaturants than soluble PC1, and more active. Both tethers allow PC1 to visit the cell surface transiently, cleaving soluble molecules outside the cell. Both membrane-bound PC1 chimeras cleave membrane PAM into soluble active fragments when PAM is expressed on adjacent cells. PMID:12084516

  19. Transportation and Accumulation of Redox Active Species at the Buried Interfaces of Plasticized Membrane Electrodes.

    PubMed

    Sohail, Manzar; De Marco, Roland; Jarolímová, Zdeňka; Pawlak, Marcin; Bakker, Eric; He, Ning; Latonen, Rose-Marie; Lindfors, Tom; Bobacka, Johan

    2015-09-29

    The transportation and accumulation of redox active species at the buried interface between glassy carbon electrodes and plasticized polymeric membranes have been studied using synchrotron radiation X-ray photoelectron spectroscopy (SR-XPS), near edge X-ray absorption fine structure (NEXAFS), in situ electrochemical Fourier transform infrared-attenuated total reflectance (FTIR-ATR) spectroscopy, cyclic voltammetry (CV), chronoamperometry (CA), and electrochemical impedance spectroscopy (EIS). Ferrocene tagged poly(vinyl chloride) [FcPVC], ferrocene (Fc), and its derivatives together with tetracyanoquinodimethane (TCNQ) doped plasticized polymeric membrane electrodes have been investigated, so as to extend the study of the mechanism of this reaction chemistry to different time scales (both small and large molecules with variable diffusion coefficients) using a range of complementary electrochemical and surface analysis techniques. This study also provides direct spectroscopic evidence for the transportation and electrochemical reactivity of redox active species, regardless of the size of the electrochemically reactive molecule, at the buried interface of the substrate electrode. With all redox dopants, when CA electrolysis was performed, redox active species were undetectable (<1 wt % of signature elements or below the detection limit of SR-XPS and NEXAFS) in the outermost surface layers of the membrane, while a high concentration of redox species was located at the electrode substrate as a consequence of the deposition of the reaction product (Fc(+)-anion complex) at the buried interface between the electrode and the membrane. This reaction chemistry for redox active species within plasticized polymeric membranes may be useful in the fashioning of multilayered polymeric devices (e.g., chemical sensors, organic electronic devices, protective laminates, etc.) based on an electrochemical tunable deposition of redox molecules at the buried substrate electrode beneath

  20. Trans-activity of plasma membrane-associated ganglioside sialyltransferase in mammalian cells.

    PubMed

    Vilcaes, Aldo A; Demichelis, Vanina Torres; Daniotti, Jose L

    2011-09-01

    Gangliosides are acidic glycosphingolipids that contain sialic acid residues and are expressed in nearly all vertebrate cells. They are synthesized at the Golgi complex by a combination of glycosyltransferase activities followed by vesicular delivery to the plasma membrane, where they participate in a variety of physiological as well as pathological processes. Recently, a number of enzymes of ganglioside anabolism and catabolism have been shown to be associated with the plasma membrane. In particular, it was observed that CMP-NeuAc:GM3 sialyltransferase (Sial-T2) is able to sialylate GM3 at the plasma membrane (cis-catalytic activity). In this work, we demonstrated that plasma membrane-integrated ecto-Sial-T2 also displays a trans-catalytic activity at the cell surface of epithelial and melanoma cells. By using a highly sensitive enzyme-linked immunosorbent assay combined with confocal fluorescence microscopy, we observed that ecto-Sial-T2 was able to sialylate hydrophobically or covalently immobilized GM3 onto a solid surface. More interestingly, we observed that ecto-Sial-T2 was able to sialylate GM3 exposed on the membrane of neighboring cells by using both the exogenous and endogenous donor substrate (CMP-N-acetylneuraminic acid) available at the extracellular milieu. In addition, the trans-activity of ecto-Sial-T2 was considerably reduced when the expression of the acceptor substrate was inhibited by using a specific inhibitor of biosynthesis of glycolipids, indicating the lipidic nature of the acceptor. Our findings provide the first direct evidence that an ecto-sialyltransferase is able to trans-sialylate substrates exposed in the plasma membrane from mammalian cells, which represents a novel insight into the molecular events that regulate the local glycosphingolipid composition. PMID:21768099

  1. Trans-activity of Plasma Membrane-associated Ganglioside Sialyltransferase in Mammalian Cells*

    PubMed Central

    Vilcaes, Aldo A.; Demichelis, Vanina Torres; Daniotti, Jose L.

    2011-01-01

    Gangliosides are acidic glycosphingolipids that contain sialic acid residues and are expressed in nearly all vertebrate cells. They are synthesized at the Golgi complex by a combination of glycosyltransferase activities followed by vesicular delivery to the plasma membrane, where they participate in a variety of physiological as well as pathological processes. Recently, a number of enzymes of ganglioside anabolism and catabolism have been shown to be associated with the plasma membrane. In particular, it was observed that CMP-NeuAc:GM3 sialyltransferase (Sial-T2) is able to sialylate GM3 at the plasma membrane (cis-catalytic activity). In this work, we demonstrated that plasma membrane-integrated ecto-Sial-T2 also displays a trans-catalytic activity at the cell surface of epithelial and melanoma cells. By using a highly sensitive enzyme-linked immunosorbent assay combined with confocal fluorescence microscopy, we observed that ecto-Sial-T2 was able to sialylate hydrophobically or covalently immobilized GM3 onto a solid surface. More interestingly, we observed that ecto-Sial-T2 was able to sialylate GM3 exposed on the membrane of neighboring cells by using both the exogenous and endogenous donor substrate (CMP-N-acetylneuraminic acid) available at the extracellular milieu. In addition, the trans-activity of ecto-Sial-T2 was considerably reduced when the expression of the acceptor substrate was inhibited by using a specific inhibitor of biosynthesis of glycolipids, indicating the lipidic nature of the acceptor. Our findings provide the first direct evidence that an ecto-sialyltransferase is able to trans-sialylate substrates exposed in the plasma membrane from mammalian cells, which represents a novel insight into the molecular events that regulate the local glycosphingolipid composition. PMID:21768099

  2. Rotenone Activates Phagocyte NADPH Oxidase through Binding to Its Membrane Subunit gp91phox

    PubMed Central

    Zhou, Hui; Zhang, Feng; Chen, Shih-heng; Zhang, Dan; Wilson, Belinda; Hong, Jau-shyong; Gao, Hui-Ming

    2011-01-01

    Rotenone, a widely used pesticide, reproduces Parkinsonism in rodents and associates with increased risk for Parkinson’s disease. We previously reported rotenone increased superoxide production through stimulating microglial phagocyte NADPH oxidase (PHOX). The present study identified a novel mechanism by which rotenone activates PHOX. Ligand-binding assay revealed that rotenone directly bound to membrane gp91phox, the catalytic subunit of PHOX; such binding was inhibited by diphenyleneiodonium, a PHOX inhibitor with a binding site on gp91phox. Functional studies showed both membrane and cytosolic subunits were required for rotenone-induced superoxide production in cell-free systems, intact phagocytes, and COS7 cells transfected with membrane subunits (gp91phox/p22phox) and cytosolic subunits (p67phox and p47phox). Rotenone-elicited extracellular superoxide release in p47phox-deficient macrophages suggested rotenone enabled to activate PHOX through a p47phox-independent mechanism. Increased membrane translocation of p67phox, elevated binding of p67phox to rotenone-treated membrane fractions, and co-immunoprecipitation of p67phox and gp91phox in rotenone-treated wild-type and p47phox-deficient macrophages indicated p67phox played a critical role in rotenone-induced PHOX activation via its direct interaction with gp91phox. Rac1, a Rho-like small GTPase, enhanced p67phox-gp91phox interaction; Rac1 inhibition decreased rotenone-elicited superoxide release. In conclusion, rotenone directly interacted with gp91phox; such an interaction triggered membrane translocation of p67phox, leading to PHOX activation and superoxide production. PMID:22094225

  3. Active Trans-Plasma Membrane Water Cycling in Yeast Is Revealed by NMR

    PubMed Central

    Zhang, Yajie; Poirier-Quinot, Marie; Springer, Charles S.; Balschi, James A.

    2011-01-01

    Plasma membrane water transport is a crucial cellular phenomenon. Net water movement in response to an osmotic gradient changes cell volume. Steady-state exchange of water molecules, with no net flux or volume change, occurs by passive diffusion through the phospholipid bilayer and passage through membrane proteins. The hypothesis is tested that plasma membrane water exchange also correlates with ATP-driven membrane transport activity in yeast (Saccharomyces cerevisiae). Longitudinal 1H2O NMR relaxation time constant (T1) values were measured in yeast suspensions containing extracellular relaxation reagent. Two-site-exchange analysis quantified the reversible exchange kinetics as the mean intracellular water lifetime (τi), where τi−1 is the pseudo-first-order rate constant for water efflux. To modulate cellular ATP, yeast suspensions were bubbled with 95%O2/5%CO2 (O2) or 95%N2/5%CO2 (N2). ATP was high during O2, and τi−1 was 3.1 s−1 at 25°C. After changing to N2, ATP decreased and τi−1 was 1.8 s−1. The principal active yeast ion transport protein is the plasma membrane H+-ATPase. Studies using the H+-ATPase inhibitor ebselen or a yeast genetic strain with reduced H+-ATPase found reduced τi−1, notwithstanding high ATP. Steady-state water exchange correlates with H+-ATPase activity. At volume steady state, water is cycling across the plasma membrane in response to metabolic transport activity. PMID:22261073

  4. Membrane anchoring of diacylglycerol-lactones substituted with rigid hydrophobic acyl domains correlates with biological activities

    PubMed Central

    Raifman, Or; Kolusheva, Sofiya; Comin, Maria J.; Kedei, Noemi; Lewin, Nancy E.; Blumberg, Peter M.; Marquez, Victor E.; Jelinek, Raz

    2009-01-01

    Summary Synthetic diacylglycerol lactones (DAG-lactones) are effective modulators of critical cellular signaling pathways, downstream of the lipophilic second messenger diacylglycerol, that activate a host of protein kinase C (PKC) isozymes as well as other non-kinase proteins that share with PKC similar C1 membrane-targeting domains. A fundamental determinant of the biological activity of these amphiphilic molecules is the nature of their interactions with cellular membranes. This study characterizes the membrane interactions and bilayer anchoring of a series of DAG-lactones in which the hydrophobic moiety is a “molecular rod”, namely a rigid 4-[2-(R-phenyl)ethynyl]benzoate moiety in the acyl position. Application of assays employing chromatic biomimetic vesicles and biophysical techniques reveals that the mode of membrane anchoring of the DAG-lactone derivatives was markedly affected by the presence of the hydrophobic diphenyl rod and by the size of the functional unit displayed at the terminus of the rod. Two primary mechanisms of interaction were observed: surface binding of the DAG-lactones at the lipid/water interface and deep insertion of the ligands into the alkyl core of the lipid bilayer. These membrane-insertion properties could explain the different patterns of PKC translocation from cytosol to membranes induced by the molecular-rod DAG-lactones. This investigation emphasizes that the side-residues of DAG-lactones, rather than simply conferring hydrophobicity, profoundly influence membrane interactions and in that fashion may further contribute to the diversity of biological actions of these synthetic biomimetic ligands. PMID:19961537

  5. tBid Undergoes Multiple Conformational Changes at the Membrane Required for Bax Activation*

    PubMed Central

    Shamas-Din, Aisha; Bindner, Scott; Zhu, Weijia; Zaltsman, Yehudit; Campbell, Clinton; Gross, Atan; Leber, Brian; Andrews, David W.; Fradin, Cécile

    2013-01-01

    Bid is a Bcl-2 family protein that promotes apoptosis by activating Bax and eliciting mitochondrial outer membrane permeabilization (MOMP). Full-length Bid is cleaved in response to apoptotic stimuli into two fragments, p7 and tBid (p15), that are held together by strong hydrophobic interactions until the complex binds to membranes. The detailed mechanism(s) of fragment separation including tBid binding to membranes and release of the p7 fragment to the cytoplasm remain unclear. Using liposomes or isolated mitochondria with fluorescently labeled proteins at physiological concentrations as in vitro models, we report that the two components of the complex quickly separate upon interaction with a membrane. Once tBid binds to the membrane, it undergoes slow structural rearrangements that result in an equilibrium between two major tBid conformations on the membrane. The conformational change of tBid is a prerequisite for interaction with Bax and is, therefore, a novel step that can be modulated to promote or inhibit MOMP. Using automated high-throughput image analysis in cells, we show that down-regulation of Mtch2 causes a significant delay between tBid and Bax relocalization in cells. We propose that by promoting insertion of tBid via a conformational change at the mitochondrial outer membrane, Mtch2 accelerates tBid-mediated Bax activation and MOMP. Thus the interaction of Mtch2 and tBid is a potential target for therapeutic control of Bid initiated cell death. PMID:23744079

  6. Plasma membrane restricted RhoGEF activity is sufficient for RhoA-mediated actin polymerization

    PubMed Central

    van Unen, Jakobus; Reinhard, Nathalie R.; Yin, Taofei; Wu, Yi I.; Postma, Marten; Gadella, Theodorus W.J.; Goedhart, Joachim

    2015-01-01

    The small GTPase RhoA is involved in cell morphology and migration. RhoA activity is tightly regulated in time and space and depends on guanine exchange factors (GEFs). However, the kinetics and subcellular localization of GEF activity towards RhoA are poorly defined. To study the mechanism underlying the spatiotemporal control of RhoA activity by GEFs, we performed single cell imaging with an improved FRET sensor reporting on the nucleotide loading state of RhoA. By employing the FRET sensor we show that a plasma membrane located RhoGEF, p63RhoGEF, can rapidly activate RhoA through endogenous GPCRs and that localized RhoA activity at the cell periphery correlates with actin polymerization. Moreover, synthetic recruitment of the catalytic domain derived from p63RhoGEF to the plasma membrane, but not to the Golgi apparatus, is sufficient to activate RhoA. The synthetic system enables local activation of endogenous RhoA and effectively induces actin polymerization and changes in cellular morphology. Together, our data demonstrate that GEF activity at the plasma membrane is sufficient for actin polymerization via local RhoA signaling. PMID:26435194

  7. Regulation of Rac1 translocation and activation by membrane domains and their boundaries

    PubMed Central

    Moissoglu, Konstadinos; Kiessling, Volker; Wan, Chen; Hoffman, Brenton D.; Norambuena, Andres; Tamm, Lukas K.; Schwartz, Martin Alexander

    2014-01-01

    ABSTRACT The activation of Rac1 and related Rho GTPases involves dissociation from Rho GDP-dissociation inhibitor proteins and translocation to membranes, where they bind effectors. Previous studies have suggested that the binding of Rac1 to membranes requires, and colocalizes with, cholesterol-rich liquid-ordered (lo) membrane domains (lipid rafts). Here, we have developed a fluorescence resonance energy transfer (FRET) assay that robustly detects Rac1 membrane targeting in living cells. Surprisingly, FRET with acceptor constructs that were targeted to either raft or non-raft areas indicated that Rac1 was present in both regions. Functional studies showed that Rac1 localization to non-raft regions decreased GTP loading as a result of inactivation by GTPase-activating proteins. In vitro, Rac1 translocation to supported lipid bilayers also required lo domains, yet Rac1 was concentrated in the liquid-disordered (ld) phase. Single-molecule analysis demonstrated that translocation occurred preferentially at lo–ld boundaries. These results, therefore, suggest that Rac1 translocates to the membrane at domain boundaries, then diffuses into raft and non-raft domains, which controls interactions. These findings resolve discrepancies in our understanding of Rac biology and identify novel mechanisms by which lipid rafts modulate Rho GTPase signaling. PMID:24695858

  8. Agonist-activated Ca2+ influx occurs at stable plasma membrane and endoplasmic reticulum junctions

    PubMed Central

    Treves, Susan; Vukcevic, Mirko; Griesser, Johanna; Armstrong, Clara-Franzini; Zhu, Michael X.; Zorzato, Fancesco

    2010-01-01

    Junctate is a 33 kDa integral protein of sarco(endo)plasmic reticulum membranes that forms a macromolecular complex with inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] receptors and TRPC3 channels. TIRF microscopy shows that junctate enhances the number of fluorescent puncta on the plasma membrane. The size and distribution of these puncta are not affected by the addition of agonists that mobilize Ca2+ from Ins(1,4,5)P3-sensitive stores. Puncta are associated with a significantly larger number of peripheral junctions between endoplasmic reticulum and plasma membrane, which are further enhanced upon stable co-expression of junctate and TRPC3. The gap between the membranes of peripheral junctions is bridged by regularly spaced electron-dense structures of 10 nm. Ins(1,4,5)P3 inhibits the interaction of the cytoplasmic N-terminus of junctate with the ligand-binding domain of the Ins(1,4,5)P3 receptor. Furthermore, Ca2+ influx evoked by activation of Ins(1,4,5)P3 receptors is increased where puncta are located. We conclude that stable peripheral junctions between the plasma membrane and endoplasmic reticulum are the anatomical sites of agonist-activated Ca2+ entry. PMID:21062895

  9. A hybrid anaerobic membrane bioreactor coupled with online ultrasonic equipment for digestion of waste activated sludge.

    PubMed

    Xu, Meilan; Wen, Xianghua; Yu, Zhiyong; Li, Yushan; Huang, Xia

    2011-05-01

    Anaerobic membrane bioreactor and online ultrasonic equipment used to enhance membrane filtration were coupled to form a hybrid system (US-AnMBR) designed for long-term digestion of waste activated sludge. The US-AnMBR was operated under volatile solids loading rates of 1.1-3.7 gVS/L·d. After comprehensive studies on digestion performance and membrane fouling control in the US-AnMBR, the final loading rate was determined to be 2.7 gVS/L·d with 51.3% volatile solids destruction. In the US-AnMBR, the improved digestion was due to enhanced sludge disintegration, as indicated by soluble matter comparison in the supernatant and particle size distribution in the digested sludge. Maximum specific methanogenic activity revealed that ultrasound application had no negative effect on anaerobic microorganisms. Furthermore, implementing ultrasound effectively controlled membrane fouling and successfully facilitated membrane bioreactor operation. This lab-scale study demonstrates the potential feasibility and effectiveness of setting up a US-AnMBR system for sludge digestion. PMID:21421308

  10. Active Curved Polymers Form Vortex Patterns on Membranes

    NASA Astrophysics Data System (ADS)

    Denk, Jonas; Huber, Lorenz; Reithmann, Emanuel; Frey, Erwin

    2016-04-01

    Recent in vitro experiments with FtsZ polymers show self-organization into different dynamic patterns, including structures reminiscent of the bacterial Z ring. We model FtsZ polymers as active particles moving along chiral, circular paths by Brownian dynamics simulations and a Boltzmann approach. Our two conceptually different methods point to a generic phase behavior. At intermediate particle densities, we find self-organization into vortex structures including closed rings. Moreover, we show that the dynamics at the onset of pattern formation is described by a generalized complex Ginzburg-Landau equation.

  11. Active Curved Polymers Form Vortex Patterns on Membranes.

    PubMed

    Denk, Jonas; Huber, Lorenz; Reithmann, Emanuel; Frey, Erwin

    2016-04-29

    Recent in vitro experiments with FtsZ polymers show self-organization into different dynamic patterns, including structures reminiscent of the bacterial Z ring. We model FtsZ polymers as active particles moving along chiral, circular paths by Brownian dynamics simulations and a Boltzmann approach. Our two conceptually different methods point to a generic phase behavior. At intermediate particle densities, we find self-organization into vortex structures including closed rings. Moreover, we show that the dynamics at the onset of pattern formation is described by a generalized complex Ginzburg-Landau equation. PMID:27176542

  12. Neuropeptide-degrading endopeptidase activity of locust (Schistocerca gregaria) synaptic membranes.

    PubMed

    Isaac, R E

    1988-11-01

    Locust adipokinetic hormone (AKH, pGlu-Leu-Asn-Phe-Thr-Pro-Asn-Trp-Gly-Thr-NH2) was used as the substrate to measure neuropeptide-degrading endopeptidase activity in neutral membranes from ganglia of the locust Schistocerca gregaria. Initial hydrolysis of AKH at neural pH by peptidases of washed neural membranes generated pGlu-Leu-Asn and Phe-Thr-Pro-Asn-Trp-Gly-Thr-NH2 as primary metabolites, demonstrating that degradation was initiated by cleavage of the Asn-Phe bond. Amastatin protected the C-terminal fragment from further metabolism by aminopeptidase activity without inhibiting AKH degradation. The same fragments were generated on incubation of AKH with purified pig kidney endopeptidase 24.11, and enzyme known to cleave peptide bonds that involve the amino group of hydrophobic amino acids. Phosphoramidon (10 microM), a selective inhibitor of mammalian endopeptidase 24.11, partially inhibited the endopeptidase activity of locust neural membranes. This phosphoramidon-sensitive activity was shown to enriched in a synaptic membrane preparation with around 80% of the activity being inhibited by 10 microM-phosphoramidon (IC50 = 0.2 microM). The synaptic endopeptidase was also inhibited by 1 mM-EDTA, 1 mM-1,10-phenanthroline and 1 microM-thiorphan, and the activity was maximal between pH 7.3 and 8.0. Localization of the phosphoramidon-sensitive enzyme in synaptic membranes is consistent with a physiological role for this endopeptidase in the metabolism of insect peptides at the synapse. PMID:3063256

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

    PubMed

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

    2015-06-01

    The membrane-active glycopeptide antibiotic YV11455 is a lipophilic cationic vancomycin analogue that demonstrates rapid and concentration-dependent killing of clinically relevant multidrug-resistant (MDR) Gram-positive bacteria in vitro. YV11455 was 2-fold and 54-270-fold more effective than vancomycin against clinical isolates of vancomycin-sensitive and vancomycin-resistant bacteria, respectively. In this study, the in vivo efficacy, pharmacodynamics, pharmacokinetics and acute toxicology of YV11455 were investigated. In vivo activity and pharmacodynamics were determined in the neutropenic mouse thigh infection model against meticillin-resistant Staphylococcus aureus (MRSA). YV11455 produced dose-dependent reductions in MRSA titres in thigh muscle. When administered intravenously, the 50% effective dose (ED(50)) for YV11455 against MRSA was found to be 3.3 mg/kg body weight, and titres were reduced by up to ca. 3log(10)CFU/g from pre-treatment values at a dosage of 12 mg/kg with single treatment. Single-dose pharmacokinetic studies demonstrated linear kinetics and a prolonged half-life, with an increase in drug exposure (area under the concentration-time curve) compared with vancomycin. The peak plasma concentration following an intravenous dose of 12 mg/kg was 543.5 μg/mL. Acute toxicology studies revealed that YV11455 did not cause any significant alterations in biochemical parameters or histological pictures related to major organs such as the liver and kidney at its pharmacodynamic endpoint (ED(3-log kill)). These findings collectively suggest that YV11455 could be used clinically for the treatment of infections caused by MDR Gram-positive bacteria. PMID:25900818

  14. Combining phosphate and bacteria removal on chemically active filter membranes allows prolonged storage of drinking water.

    PubMed

    Rotzetter, A C C; Kellenberger, C R; Schumacher, C M; Mora, C; Grass, R N; Loepfe, M; Luechinger, N A; Stark, W J

    2013-11-13

    A chemically active filtration membrane with incorporated lanthanum oxide nanoparticles enables the removal of bacteria and phosphate at the same time and thus provides a simple device for preparation of drinking water and subsequent safe storage without using any kind of disinfectants. PMID:23913409

  15. A novel fragment based strategy for membrane active antimicrobials against MRSA.

    PubMed

    Li, Jianguo; Liu, Shouping; Koh, Jun-Jie; Zou, Hanxun; Lakshminarayanan, Rajamani; Bai, Yang; Pervushin, Konstantin; Zhou, Lei; Verma, Chandra; Beuerman, Roger W

    2015-04-01

    Membrane active antimicrobials are a promising new generation of antibiotics that hold the potential to avert antibiotic resistance. However, poor understanding of the action mechanism and the lack of general design principles have impeded their development. Here we extend the concept of fragment based drug design and propose a pharmacophore model based on first principles for the design of membrane active antimicrobials against Gram positive pathogens. Elaborating on a natural xanthone-based hydrophobic scaffold, two derivatives of the pharmacophore model are proposed, and these demonstrate excellent antimicrobial activity. Rigorous molecular dynamics simulations combined with biophysical experiments suggest a three-step mechanism of action (absorption-translocation-disruption) which allows us to identify key factors for the practical optimization of each fragment of the pharmacophore. Moreover, the model matches the structures of several membrane active antimicrobials which are currently in clinical trials. Our model provides a novel and rational approach for the design of bactericidal molecules that target the bacterial membrane. PMID:25582665

  16. Antibacterial Activity of Shikimic Acid from Pine Needles of Cedrus deodara against Staphylococcus aureus through Damage to Cell Membrane

    PubMed Central

    Bai, Jinrong; Wu, Yanping; Liu, Xiaoyan; Zhong, Kai; Huang, Yina; Gao, Hong

    2015-01-01

    Shikimic acid (SA) has been reported to possess antibacterial activity against Staphylococcus aureus, whereas the mode of action of SA is still elusive. In this study, the antibacterial activity and mechanism of SA toward S. aureus by cell membrane damage was investigated. After SA treatment, massive K+ and nucleotide leakage from S. aureus, and a significant change in the membrane potential was observed, suggesting SA may act on the membrane by destroying the cell membrane permeability. Through transmission electron microscopic observations we further confirmed that SA can disrupt the cell membrane and membrane integrity. Meanwhile, SA was found to be capable of reducing the membrane fluidity of the S. aureus cell. Moreover, the fluorescence experiments indicated that SA could quench fluorescence of Phe residues of the membrane proteins, thus demonstrating that SA can bind to S. aureus membrane proteins. Therefore, these results showed the antibacterial activity of SA against S. aureus could be caused by the interactions of SA with S. aureus membrane proteins and lipids, resulting in causing cell membrane dysfunction and bacterial damage or even death. This study reveals the potential use of SA as an antibacterial agent. PMID:26580596

  17. T Lymphocyte Activation Threshold and Membrane Reorganization Perturbations in Unique Culture Model

    NASA Technical Reports Server (NTRS)

    Adams, C. L.; Sams, C. F.

    2000-01-01

    Quantitative activation thresholds and cellular membrane reorganization are mechanisms by which resting T cells modulate their response to activating stimuli. Here we demonstrate perturbations of these cellular processes in a unique culture system that non-invasively inhibits T lymphocyte activation. During clinorotation, the T cell activation threshold is increased 5-fold. This increased threshold involves a mechanism independent of TCR triggering. Recruitment of lipid rafts to the activation site is impaired during clinorotation but does occur with increased stimulation. This study describes a situation in which an individual cell senses a change in its physical environment and alters its cell biological behavior.

  18. Formulation and process optimization of multiparticulate pulsatile system delivered by osmotic pressure-activated rupturable membrane.

    PubMed

    Hung, Sheng-Feng; Hsieh, Chien-Ming; Chen, Ying-Chen; Lin, Cheng-Mao; Ho, Hsiu-O; Sheu, Ming-Thau

    2015-03-01

    In this study, a multiparticulate pulsatile drug delivery system activated by a rupturable controlled-release membrane (Eudragit(®) RS) via osmotic pressure (with NaCl as the osmogent) was developed and characterized for omeprazole, omeprazole sodium, and propranolol HCl which have different water solubilities. Multiparticulates in pellet form for incorporation with or without the osmogent were manufactured by three methods and then used to coat a polymeric membrane. Results demonstrated that drug/osmogent-containing pellets manufactured by the extrusion/spheronization method with incorporation of the osmogent were optimal. The lag time (tL) to initiate pulsatile release is regulated by tL=l(2)/(6×D), which is dependent on the coating levels (l(2)) and plasticizer content (D). The pulsatile release pattern was found to be dependent on the osmotic pressure (osmogent), drug solubility, and mechanical properties of the polymeric membrane (elasticity and toughness). Omeprazole with lower water solubility could not generate sufficient osmotic pressure to create a crack in the membrane to activate pulsatile release, whereas the two other model drugs with higher solubilities could. But adsorption of omeprazole sodium on Eudragit(®) RS via charge-charge interactions led the its incomplete release. Finally, with 4% osmogent of NaCl added, a lag time in a range from 0 to 12h proportionally regulated by varying both the membrane thickness and plasticizer level initiated the complete pulsatile release of propranolol HCl. In conclusion, a multiparticulate pulsatile drug delivery system activated by a rupturable controlled-release membrane via osmotic pressure was successfully developed, and clinical applications of chronotherapy with drugs like propranolol HCl are expected. PMID:25575473

  19. Polyamines cause plasma membrane depolarization, activate Ca2+-, and modulate H+-ATPase pump activity in pea roots.

    PubMed

    Pottosin, Igor; Velarde-Buendía, Ana María; Bose, Jayakumar; Fuglsang, Anja T; Shabala, Sergey

    2014-06-01

    Polyamines regulate a variety of cation and K(+) channels, but their potential effects on cation-transporting ATPases are underexplored. In this work, noninvasive microelectrode ion flux estimation and conventional microelectrode techniques were applied to study the effects of polyamines on Ca(2+) and H(+) transport and membrane potential in pea roots. Externally applied spermine or putrescine (1mM) equally activated eosin yellow (EY)-sensitive Ca(2+) pumping across the root epidermis and caused net H(+) influx or efflux. Proton influx induced by spermine was suppressed by EY, supporting the mechanism in which Ca(2+) pump imports 2 H(+) per each exported Ca(2+). Suppression of the Ca(2+) pump by EY diminished putrescine-induced net H(+) efflux instead of increasing it. Thus, activities of Ca(2+) and H(+) pumps were coupled, likely due to the H(+)-pump inhibition by intracellular Ca(2+). Additionally, spermine but not putrescine caused a direct inhibition of H(+) pumping in isolated plasma membrane vesicles. Spermine, spermidine, and putrescine (1mM) induced membrane depolarization by 70, 50, and 35 mV, respectively. Spermine-induced depolarization was abolished by cation transport blocker Gd(3+), was insensitive to anion channels' blocker niflumate, and was dependent on external Ca(2+). Further analysis showed that uptake of polyamines but not polyamine-induced cationic (K(+)+Ca(2+)+H(+)) fluxes were a main cause of membrane depolarization. Polyamine increase is a common component of plant stress responses. Activation of Ca(2+) efflux by polyamines and contrasting effects of polyamines on net H(+) fluxes and membrane potential can contribute to Ca(2+) signalling and modulate a variety of transport processes across the plasma membrane under stress. PMID:24723394

  20. Large aperture ac interferometer for optical testing.

    PubMed

    Moore, D T; Murray, R; Neves, F B

    1978-12-15

    A 20-cm clear aperture modified Twyman-Green interferometer is described. The system measures phase with an AC technique called phase-lock interferometry while scanning the aperture with a dual galvanometer scanning system. Position information and phase are stored in a minicomputer with disk storage. This information is manipulated with associated software, and the wavefront deformation due to a test component is graphically displayed in perspective and contour on a CRT terminal. PMID:20208642

  1. New LGS for large aperture telescope

    NASA Astrophysics Data System (ADS)

    Lukin, Vladimir; Bolbasova, Lidia

    2007-10-01

    The image quality is analyzed of an extraterrestrial object formed by astronomical optical system through the turbulent atmosphere. Relative increase the Strehl parameter is calculated under adaptive correction based on the laser guide star technique. The efficiency of adaptive correction of distortions for different type of the guide sources is compared. A special wave front sensor is applied, which operates using the broad laser beam as a reference wave. The calculations are performed for different models of the vertical variations of the structural parameter of the refractive index of the turbulent atmosphere. The wave front sensor was used, which enables to reconstruct the continuous phase of the reference wave. As the estimates show, the parameters of the formed field are quite close to that plane wave. So the higher correction and big increase of the Strehl parameter are obtained, that is indirect evidence of the good correction of the higher mode components, which are badly corrected using the traditional techniques for formation of LGS by means of a focused laser beam. As comparative calculations for different models of vertical variations of the structural parameter of the refractive index have shown, there are serious differences in the behaviors of the correlation radii for the plane and spherical waves.

  2. Large Aperture Multiplexed Diffractive Lidar Optics

    NASA Technical Reports Server (NTRS)

    Rallison, Richard D.; Schwemmer, Geary K. (Technical Monitor)

    1999-01-01

    We have delivered only 2 or 3 UV Holographic Optical Elements (HOEs) thus far and have fallen short of the intended goal in size and in dual wavelength function. Looking back, it has been fortuitous that we even made anything work in the UV region. It was our good fortune to discover that the material we work with daily was adequate for use at 355 nm, if well rinsed during processing. If we had stuck to our original plan of etching in small pieces of fused silica, we would still be trying to make the first small section in our ion mill, which is not yet operational. The original plan was far too ambitious and would take another 2 years to complete beginning where we left off this time. In order to make a HOE for the IR as well as the UV we will likely have to learn to sensitize some film to the 1064 line and we have obtained sensitizer that is reported to work in that region already. That work would also take an additional year to complete.

  3. Large Aperture Systems: 2000-2004

    NASA Technical Reports Server (NTRS)

    2004-01-01

    This custom bibliography from the NASA Scientific and Technical Information Program lists a sampling of records found in the NASA Aeronautics and Space Database. The scope of this topic includes technologies for next generation astronomical telescopes and detectors. This area of focus is one of the enabling technologies as defined by NASA s Report of the President s Commission on Implementation of United States Space Exploration Policy, published in June 2004.

  4. Dielectric elastomer laminates for active membrane pump applications

    NASA Astrophysics Data System (ADS)

    Pope, Kimberly; Tews, Alyson; Frecker, Mary I.; Mockensturm, Eric; Goulbourne, Nakhiah C.; Snyder, Alan J.

    2004-07-01

    Previous research has demonstrated promise for the use of dielectric elastomer (DE) films in diaphragm pump applications. Because the films tend to be quite thin, single layers operate at very low pressures. To make this technology suitable for practical applications, the films may be organized into laminates which will operate at increased pressures. Radially stretched circular diaphragms of two materials were tested: 3M VHB 4905 polyacrylate and spin-cast Nusil CF19-2186 silicone. The diaphragms were stacked, each layer sharing an electrode with the adjacent layer. The stack was mounted on a sealed chamber and energized at varied electric fields while regulated pressure was applied to the interior chamber, displacing the diaphragm. The pressure-volume properties of the stacks were recorded for each activation state.

  5. Active Motion of Hair Bundles Coupled to the Otolithic Membrane in the Frog Sacculus

    NASA Astrophysics Data System (ADS)

    Strimbu, C. Elliott; Fredrickson-Hemsing, Lea; Bozovic, Dolores

    2011-11-01

    Active hair bundle motility has been proposed to provide the basis for the active process in the auditory organs of non-mammalian vertibrates, and has been extensively studied in mechanically decoupled or free-standing hair bundles from in vitro preparations of the frog sacculus. A number of studies have, however, suggested that cooperativity between hair cells plays an important role in the response of an intact organ. We use a semi-intact in vitro saccular preparation in which the hair cells are coupled and loaded by the otolithic membrane. While the hair bundles do not spontaneously oscillate beneath the membrane, they exhibit active movements in response to transient stimuli, demonstrating that the active process remains operant under these conditions. The coupled system however displays a striking decrease in frequency selectivity compared to freely oscillating bundles.

  6. Effect of low dosages of powdered activated carbon on membrane bioreactor performance.

    PubMed

    Remy, Maxime; Temmink, Hardy; Rulkens, Wim

    2012-01-01

    Previous research has demonstrated that powdered activated carbon (PAC), when applied at very low dosages and long SRTs, reduces membrane fouling in membrane bioreactors (MBRs). This effect was related to the formation of stronger sludge flocs, which are less sensitive to shear. In this contribution the long-term effect of PAC addition was studied by running two parallel MBRs on sewage. To one of these, PAC was dosed and a lower fouling tendency of the sludge was verified, with a 70% longer sustainable filtration time. Low PAC dosages showed additional advantages with regard to oxygen transfer and dewaterability, which may provide savings on operational costs. PMID:22339033

  7. Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores.

    PubMed

    Liu, Xing; Zhang, Zhibin; Ruan, Jianbin; Pan, Youdong; Magupalli, Venkat Giri; Wu, Hao; Lieberman, Judy

    2016-07-01

    Inflammatory caspases (caspases 1, 4, 5 and 11) are activated in response to microbial infection and danger signals. When activated, they cleave mouse and human gasdermin D (GSDMD) after Asp276 and Asp275, respectively, to generate an N-terminal cleavage product (GSDMD-NT) that triggers inflammatory death (pyroptosis) and release of inflammatory cytokines such as interleukin-1β. Cleavage removes the C-terminal fragment (GSDMD-CT), which is thought to fold back on GSDMD-NT to inhibit its activation. However, how GSDMD-NT causes cell death is unknown. Here we show that GSDMD-NT oligomerizes in membranes to form pores that are visible by electron microscopy. GSDMD-NT binds to phosphatidylinositol phosphates and phosphatidylserine (restricted to the cell membrane inner leaflet) and cardiolipin (present in the inner and outer leaflets of bacterial membranes). Mutation of four evolutionarily conserved basic residues blocks GSDMD-NT oligomerization, membrane binding, pore formation and pyroptosis. Because of its lipid-binding preferences, GSDMD-NT kills from within the cell, but does not harm neighbouring mammalian cells when it is released during pyroptosis. GSDMD-NT also kills cell-free bacteria in vitro and may have a direct bactericidal effect within the cytosol of host cells, but the importance of direct bacterial killing in controlling in vivo infection remains to be determined. PMID:27383986

  8. Active urea transport and an unusual basolateral membrane composition in the gills of a marine elasmobranch.

    PubMed

    Fines, G A; Ballantyne, J S; Wright, P A

    2001-01-01

    In elasmobranch fishes, urea occurs at high concentrations (350-600 mM) in the body fluids and tissues, where it plays an important role in osmoregulation. Retention of urea by the gill against this huge blood-to-water diffusion gradient requires specialized adaptations to the epithelial cell membranes. Experiments were performed to determine the mechanisms and structural features that facilitate urea retention by the gill of the spiny dogfish Squalus acanthias. Analysis of urea uptake by gill basolateral membrane vesicles revealed the presence of a phloretin-sensitive (half inhibition 0.09 mM), sodium-coupled, secondary active urea transporter (Michaelis constant = 10.1 mM, maximal velocity = 0.34 micromol. h(-1). mg protein(-1)). We propose that this system actively transports urea out of the gill epithelial cells back into the blood against the urea concentration gradient. Lipid analyses of the basolateral membrane revealed high levels of cholesterol contributing to the highest reported cholesterol-to-phospholipid molar ratio (3.68). This unique combination of active urea transport and modification of the phospholipid bilayer membrane is responsible for decreasing the gill permeability to urea and facilitating urea retention by the gill of Squalus acanthias. PMID:11124129

  9. Activation of Membrane NADPH Oxidase Associated with Lysosome-Targeted Acid Sphingomyelinase in Coronary Endothelial Cells

    PubMed Central

    Bao, Jun-Xiang; Jin, Si; Zhang, Fan; Wang, Zheng-Chao; Li, Ningjun

    2010-01-01

    Abstract This study explored the mechanism mediating the aggregation of membrane NADPH oxidase (NOX) subunits and subsequent activation of this enzyme in bovine coronary arterial endothelial cells (CAECs). With confocal microscopy, we found that FasL stimulated lipid rafts (LRs) clustering with NOX subunit aggregation and acid sphingomyelinase (ASM) gathering, which was blocked by the siRNA of sortilin, an intracellular protein responsible for the binding and targeting of ASM to lysosomes. Correspondingly, FasL-induced O2·− production through NOX in LRs fractions was abolished by sortilin siRNA. Further, with flow-cytometry and fluorescence resonance energy transfer (FRET) analysis, we surprisingly demonstrated that after FasL stimulation, sortilin was exposed to cell membranes from lysosomes together with Lamp-1 and ASM, and these lysosomal components were aggregated and form a signaling complex in cell membranes. With co-immunoprecipitation, lysosomal sortilin and ASM were found to interact more strongly when CAECs were stimulated by FasL. Functionally, inhibition of either sortilin expression, lysosome function, LRs clustering, or NOX activity significantly attenuated FasL-induced decrease in nitric oxide (NO) levels. It is concluded that lysosome-targeted ASM, through sortilin, is able to traffic to and expose to cell-membrane surface, which may lead to LRs clustering and NOX activation in CAECs. Antioxid. Redox Signal. 12, 703–712. PMID:19761405

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

    PubMed

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

    2011-01-01

    In this study, a novel continuous reactor has been developed to produce high quality methyl esters (biodiesel) from palm oil. A microporous TiO2/Al2O3 membrane was packed with potassium hydroxide catalyst supported on palm shell activated carbon. The central composite design (CCD) of response surface methodology (RSM) was employed to investigate the effects of reaction temperature, catalyst amount and cross flow circulation velocity on the production of biodiesel in the packed bed membrane reactor. The highest conversion of palm oil to biodiesel in the reactor was obtained at 70 °C employing 157.04 g catalyst per unit volume of the reactor and 0.21 cm/s cross flow circulation velocity. The physical and chemical properties of the produced biodiesel were determined and compared with the standard specifications. High quality palm oil biodiesel was produced by combination of heterogeneous alkali transesterification and separation processes in the packed bed membrane reactor. PMID:20888219

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

    SciTech Connect

    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 M 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.

  12. Treatment of coal gasification wastewater by membrane bioreactor hybrid powdered activated carbon (MBR–PAC) system.

    PubMed

    Jia, Shengyong; Han, Hongjun; Hou, Baolin; Zhuang, Haifeng; Fang, Fang; Zhao, Qian

    2014-12-01

    A laboratory-scale membrane bioreactor hybrid powdered activated carbon (MBR–PAC) system was developed to treat coal gasification wastewater to enhance the COD, total phenols (TPh), NH4+ removals and migrate the membrane fouling. Since the MBR–PAC system operated with PAC dosage of 4 g L−1, the maximum removal efficiencies of COD, TPh and NH4+ reached 93%, 99% and 63%, respectively with the corresponding influent concentrations of 2.27 g L−1, 497 mg L−1 and 164 mg N L−1; the PAC extraction efficiencies of COD, TPh and NH4+ were 6%, 3% and 13%, respectively; the transmembrane pressure decreased 34% with PAC after 50 d operation. The results demonstrate that PAC played a key role in the enhancement of biodegradability and mitigation of membrane fouling. PMID:25461944

  13. PVDF-HFP/ether-modified polysiloxane membranes obtained via airbrush spraying as active separators for application in lithium ion batteries.

    PubMed

    Seidel, S M; Jeschke, S; Vettikuzha, P; Wiemhöfer, H-D

    2015-08-01

    Improved hybrid polymer electrolyte membranes are introduced based on ether-modified polysiloxanes and poly(vinylidene fluoride-co-hexafluoropropylene) yielding a safe separator membrane, which is able to be sprayed directly onto lithium ion battery active materials, with an active role for enhanced ion transport. PMID:26121633

  14. Gene Recruitment of the Activated INO1 Locus to the Nuclear Membrane

    PubMed Central

    2004-01-01

    The spatial arrangement of chromatin within the nucleus can affect reactions that occur on the DNA and is likely to be regulated. Here we show that activation of INO1 occurs at the nuclear membrane and requires the integral membrane protein Scs2. Scs2 antagonizes the action of the transcriptional repressor Opi1 under conditions that induce the unfolded protein response (UPR) and, in turn, activate INO1. Whereas repressed INO1 localizes throughout the nucleoplasm, the gene is recruited to the nuclear periphery upon transcriptional activation. Recruitment requires the transcriptional activator Hac1, which is produced upon induction of the UPR, and is constitutive in a strain lacking Opi1. Artificial recruitment of INO1 to the nuclear membrane permits activation in the absence of Scs2, indicating that the intranuclear localization of a gene can profoundly influence its mechanism of activation. Gene recruitment to the nuclear periphery, therefore, is a dynamic process and appears to play an important regulatory role. PMID:15455074

  15. Antibacterial and leishmanicidal activities of temporin-SHd, a 17-residue long membrane-damaging peptide.

    PubMed

    Abbassi, Feten; Raja, Zahid; Oury, Bruno; Gazanion, Elodie; Piesse, Christophe; Sereno, Denis; Nicolas, Pierre; Foulon, Thierry; Ladram, Ali

    2013-02-01

    Temporins are a family of short antimicrobial peptides (8-17 residues) that mostly show potent activity against Gram-positive bacteria. Herein, we demonstrate that temporin-SHd, a 17-residue peptide with a net charge of +2 (FLPAALAGIGGILGKLF(amide)), expressed a broad spectrum of antimicrobial activity. This peptide displayed potent antibacterial activities against Gram-negative and Gram-positive bacteria, including multi-drug resistant Staphylococcus aureus strains, as well as antiparasitic activity against promastigote and the intracellular stage (amastigote) of Leishmania infantum, at concentration not toxic for the macrophages. Temporin-SHd that is structured in a non-amphipathic α-helix in anionic membrane-mimetic environments, strongly and selectively perturbs anionic bilayer membranes by interacting with the polar head groups and acyl region of the phospholipids, with formation of regions of two coexisting phases: one phase rich in peptide and the other lipid-rich. The disruption of lipid packing within the bilayer may lead to the formation of transient pores and membrane permeation/disruption once a threshold peptide accumulation is reached. To our knowledge, Temporin-SHd represents the first known 17-residue long temporin expressing such broad spectrum of antimicrobial activity including members of the trypanosomatidae family. Additionally, since only a few shorter members (13 residues) of the temporin family are known to display antileishmanial activity (temporins-TA, -TB and -SHa), SHd is an interesting tool to analyze the antiparasitic mechanism of action of temporins. PMID:23116712

  16. Membrane Curvature-sensing and Curvature-inducing Activity of Islet Amyloid Polypeptide and Its Implications for Membrane Disruption.

    PubMed

    Kegulian, Natalie C; Sankhagowit, Shalene; Apostolidou, Melania; Jayasinghe, Sajith A; Malmstadt, Noah; Butler, Peter C; Langen, Ralf

    2015-10-23

    Islet amyloid polypeptide (IAPP) is a 37-amino acid amyloid protein intimately associated with pancreatic islet β-cell dysfunction and death in type II diabetes. In this study, we combine spectroscopic methods and microscopy to investigate α-helical IAPP-membrane interactions. Using light scattering and fluorescence microscopy, we observe that larger vesicles become smaller upon treatment with human or rat IAPP. Electron microscopy shows the formation of various highly curved structures such as tubules or smaller vesicles in a membrane-remodeling process, and spectrofluorometric detection of vesicle leakage shows disruption of membrane integrity. This effect is stronger for human IAPP than for the less toxic rat IAPP. From CD spectra in the presence of different-sized vesicles, we also uncover the membrane curvature-sensing ability of IAPP and find that it transitions from inducing to sensing membrane curvature when lipid negative charge is decreased. Our in vivo EM images of immunogold-labeled rat IAPP and human IAPP show both forms to localize to mitochondrial cristae, which contain not only locally curved membranes but also phosphatidylethanolamine and cardiolipin, lipids with high spontaneous negative curvature. Disruption of membrane integrity by induction of membrane curvature could apply more broadly to other amyloid proteins and be responsible for membrane damage observed in other amyloid diseases as well. PMID:26283787

  17. Scolopendin 2, a cationic antimicrobial peptide from centipede, and its membrane-active mechanism.

    PubMed

    Lee, Heejeong; Hwang, Jae-Sam; Lee, Jaeho; Kim, Jae Il; Lee, Dong Gun

    2015-02-01

    Scolopendin 2 is a 16-mer peptide (AGLQFPVGRIGRLLRK) derived from the centipede Scolopendra subspinipes mutilans. We observed that this peptide exhibited antimicrobial activity in a salt-dependent manner against various fungal and bacterial pathogens and showed no hemolytic effect in the range of 1.6 μM to 100 μM. Circular dichroism analysis showed that the peptide has an α-helical properties. Furthermore, we determined the mechanism(s) of action using flow cytometry and by investigating the release of intracellular potassium. The results showed that the peptide permeabilized the membranes of Escherichia coli O157 and Candida albicans, resulting in loss of intracellular potassium ions. Additionally, bis-(1,3-dibutylbarbituric acid) trimethine oxonol and 3,3'-dipropylthiacarbocyanine iodide assays showed that the peptide caused membrane depolarization. Using giant unilamellar vesicles encapsulating calcein and large unilamellar vesicles containing fluorescein isothiocyanate-dextran, which were similar in composition to typical E. coli O157 and C. albicans membranes, we demonstrated that scolopendin 2 disrupts membranes, resulting in a pore size between 4.8 nm and 5.0 nm. Thus, we have demonstrated that a cationic antimicrobial peptide, scolopendin 2, exerts its broad-spectrum antimicrobial effects by forming pores in the cell membrane. PMID:25462167

  18. Effect of organic solvents on nervous cell membrane as measured by changes in the (Ca2+/Mg2+) ATPase activity and fluidity of synaptosomal membrane.

    PubMed

    Edelfors, S; Ravn-Jonsen, A

    1992-03-01

    The effect of various solvents on the central nervous system was studied by using rat brain synaptosomal membranes as an in vitro model. The activity of (Ca2+/Mg2+) ATPase and the membrane fluidity was determined. The alteration of the ATPase activity depended on the physio-chemical characteristics of the solvent in question. Incubation with aliphatic alkanes caused a stimulation of the ATPase activity whereas mixed hydrocarbons as kerosene, white spirit and gasoline inhibited the enzyme. Incubation with chlorinated hydrocarbons caused a biphasic response dependent on the concentration. Oxygen-containing hydrocarbons exhibited various effects as found after incubation with hydrocarbons. The different effects of the solvents on the ATPase activity suggest that the lipophilicity of the solvents is one of more parameters affecting the membrane. Furthermore, the biphasic response following the incubation with chlorinated hydrocarbons indicates that more mechanisms are involved in the enzyme effect. The membrane fluidity is increased with higher concentrations of the solvents. From the results it is concluded that the ATPase activity depends not only on the membrane fluidity and volume, but also on the hydrophilic vicinity of the enzyme molecule. PMID:1533717

  19. Age characteristics of changes in invertase activity of the mucous membrane of the small intestine

    NASA Technical Reports Server (NTRS)

    Rakhimov, K. R.; Aleksandrova, N. V.

    1980-01-01

    Rats of varying ages were subjected to stress from heat, cold, and hydrocortisone injection. Invertase activity in homogenates of small intestine mucous membranes was studied following sacrifice. Invertase activity was low in young animals, but increased sharply in 30 day old ones, remaining at a relatively constant level until old age. The study concludes that the stress hormone (corticosteroids, etc.) levels in the blood, which affects the formation of enteric enzyme levels and activities, and that age related peculiarities in invertase activity are a consequence of altered hormone status and epitheliocyte sensitivity.

  20. Cytotoxic bile acids, but not cytoprotective species, inhibit the ordering effect of cholesterol in model membranes at physiologically active concentrations.

    PubMed

    Mello-Vieira, João; Sousa, Tânia; Coutinho, Ana; Fedorov, Aleksander; Lucas, Susana D; Moreira, Rui; Castro, Rui E; Rodrigues, Cecília M P; Prieto, Manuel; Fernandes, Fábio

    2013-09-01

    Submillimolar concentrations of cytotoxic bile acids (BAs) induce cell death via apoptosis. On the other hand, several cytoprotective BAs were shown to prevent apoptosis in the same concentration range. Still, the mechanisms by which BAs trigger these opposite signaling effects remain unclear. This study was aimed to determine if cytotoxic and cytoprotective BAs, at physiologically active concentrations, are able to modulate the biophysical properties of lipid membranes, potentially translating into changes in the apoptotic threshold of cells. Binding of BAs to membranes was assessed through the variation of fluorescence parameters of suitable derivatized BAs. These derivatives partitioned with higher affinity to liquid disordered than to the cholesterol-enriched liquid ordered domains. Unlabeled BAs were also shown to have a superficial location upon interaction with the lipid membrane. Additionally, the interaction of cytotoxic BAs with membranes resulted in membrane expansion, as concluded from FRET data. Moreover, it was shown that cytotoxic BAs were able to significantly disrupt the ordering of the membrane by cholesterol at physiologically active concentrations of the BA, an effect not associated with cholesterol removal. On the other hand, cytoprotective bile acids had no effect on membrane properties. It was concluded that, given the observed effects on membrane rigidity, the apoptotic activity of cytotoxic BAs could be potentially associated with changes in plasma membrane organization (e.g. modulation of lipid domains) or with an increase in mitochondrial membrane affinity for apoptotic proteins. PMID:23747364

  1. Phospholipid base exchange activity in the leukocyte membranes of patients with inflammatory disorders.

    PubMed Central

    Niwa, Y.; Sakane, T.; Ozaki, Y.; Kanoh, T.; Taniguchi, S.

    1987-01-01

    Phospholipid base exchange and cholinephosphotransferase (CPT) and ethanolaminephosphotransferase (EPT) activities were assessed in the membranes of neutrophils or lymphocytes from patients with various inflammatory disorders. Ethanolamine exchange activity was significantly enhanced in both neutrophils and lymphocytes from patients with active Behçet's disease, active systemic lupus erythematosus (SLE), and severe bacterial infections and slightly enhanced in those from patients with active rheumatoid arthritis (RA), compared with healthy controls. No abnormal findings were found in CPT, EPT, or serine or choline base exchange activities in the leukocytes from any of the diseased groups tested or in the ethanolamine exchange activity of patients with severe viral infections and inactive SLE, RA, and Behçet's disease. The authors have recently demonstrated the enhancement of transmethylation and phospholipase A2 activity in human leukocyte membranes at the height of inflammatory disease states, as well as the activation of leukocyte ethanolamine exchange by bioactive stimulants. These data postulate that phosphatidylethanolamine synthesis by the base exchange reaction may be the precursor of transmethylation and its subsequent activation of phospholipase A2, leading to the induction of arachidonic acid cascade. PMID:3034067

  2. Measuring the activity of heterotrophic microorganism in membrane bioreactor for drinking water treatment.

    PubMed

    Han, Zheng-Shuang; Tian, Jia-Yu; Liang, Heng; Ma, Jun; Yu, Hua-Rong; Li, Kai; Ding, An; Li, Gui-Bai

    2013-02-01

    In order to quantify the activity of heterotrophic microorganism in membrane bioreactor (MBR) for drinking water treatment, biomass respiration potential (BRP) test and 2,3,5-triphenyl tetrazolium chloride-dehydrogenase activity (TTC-DHA) test were introduced and modified. A sludge concentration ratio of 5:1, incubation time of 2h, an incubation temperature that was close to the real operational temperature, and using a mixture of main AOC components as the substrate were adopted as the optimum parameters for determination of DHA in drinking water MBR. A remarkable consistency among BDOC removal, BRP and DHA for assessing biological performance in different MBRs was achieved. Moreover, a significant correlation between the BRP and DHA results of different MBRs was obtained. However, the TTC-DHA test was expected to be inaccurate for quantifying the biomass activity in membrane adsorption bioreactor (MABR), while the BRP test turned out to be still feasible in that case. PMID:23306121

  3. New protein kinase from plasma membrane of Ehrlich ascites tumor cells activated by natural polypeptides.

    PubMed Central

    Racker, E; Abdel-Ghany, M; Sherrill, K; Riegler, C; Blair, E A

    1984-01-01

    A polypeptide-dependent protein kinase was purified about 80-fold from an extract of plasma membranes of Ehrlich ascites tumor cells. The membranes were extracted with Nonidet P-40, and the extract was purified by ammonium sulfate fractionation and hydroxylapatite and affinity chromatography. The activity was stimulated 10-fold or more by polypeptide preparations from a variety of tissues, including placenta and hypothalamus. Polypeptide-dependent protein kinase had a pH optimum of about 7.5 and required Mg2+ for activity. Mn2+ at low concentrations (200 microM) stimulated enzyme activity somewhat but inhibited activity strongly at higher concentrations. The best available substrate for polypeptide-dependent protein kinase was beta-casein, and little or no phosphorylation was observed with alpha-casein, kappa-casein, phosvitin, alpha-lactalbumin, alpha-lactoglobulin, and histone. However, several endogenous substrates from plasma membranes of Ehrlich ascites tumor cells were phosphorylated. Polypeptide-dependent protein kinase activity was not inhibited by 10 mM N-ethylmaleimide, and this resistance was useful in differentiating this protein kinase from other protein kinases that were present in crude fractions and sensitive to the inhibitor. Images PMID:6589591

  4. Clathrin and Membrane Microdomains Cooperatively Regulate RbohD Dynamics and Activity in Arabidopsis.

    PubMed

    Hao, Huaiqing; Fan, Lusheng; Chen, Tong; Li, Ruili; Li, Xiaojuan; He, Qihua; Botella, Miguel A; Lin, Jinxing

    2014-04-22

    Arabidopsis thaliana respiratory burst oxidase homolog D (RbohD) functions as an essential regulator of reactive oxygen species (ROS). However, our understanding of the regulation of RbohD remains limited. By variable-angle total internal reflection fluorescence microscopy, we demonstrate that green fluorescent protein (GFP)-RbohD organizes into dynamic spots at the plasma membrane. These RbohD spots have heterogeneous diffusion coefficients and oligomerization states, as measured by photobleaching techniques. Stimulation with ionomycin and calyculin A, which activate the ROS-producing enzymatic activity of RbohD, increases the diffusion and oligomerization of RbohD. Abscisic acid and flg22 treatments also increase the diffusion coefficient and clustering of GFP-RbohD. Single-particle analysis in clathrin heavy chain2 mutants and a Flotillin1 artificial microRNA line demonstrated that clathrin- and microdomain-dependent endocytic pathways cooperatively regulate RbohD dynamics. Under salt stress, GFP-RbohD assembles into clusters and then internalizes into the cytoplasm. Dual-color fluorescence cross-correlation spectroscopy analysis further showed that salt stress stimulates RbohD endocytosis via membrane microdomains. We demonstrate that microdomain-associated RbohD spots diffuse at the membrane with high heterogeneity, and these dynamics closely relate to RbohD activity. Our results provide insight into the regulation of RbohD activity by clustering and endocytosis, which facilitate the activation of redox signaling pathways. PMID:24755455

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

    SciTech Connect

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

    2014-11-15

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

  6. Calcium release through P2X4 activates calmodulin to promote endolysosomal membrane fusion

    PubMed Central

    Cao, Qi; Zhong, Xi Zoë; Zou, Yuanjie; Murrell-Lagnado, Ruth; Zhu, Michael X.

    2015-01-01

    Intra-endolysosomal Ca2+ release is required for endolysosomal membrane fusion with intracellular organelles. However, the molecular mechanisms for intra-endolysosomal Ca2+ release and the downstream Ca2+ targets involved in the fusion remain elusive. Previously, we demonstrated that endolysosomal P2X4 forms channels activated by luminal adenosine triphosphate in a pH-dependent manner. In this paper, we show that overexpression of P2X4, as well as increasing endolysosomal P2X4 activity by alkalinization of endolysosome lumen, promoted vacuole enlargement in cells and endolysosome fusion in a cell-free assay. These effects were prevented by inhibiting P2X4, expressing a dominant-negative P2X4 mutant, and disrupting the P2X4 gene. We further show that P2X4 and calmodulin (CaM) form a complex at endolysosomal membrane where P2X4 activation recruits CaM to promote fusion and vacuolation in a Ca2+-dependent fashion. Moreover, P2X4 activation-triggered fusion and vacuolation were suppressed by inhibiting CaM. Our data thus suggest a new molecular mechanism for endolysosomal membrane fusion involving P2X4-mediated endolysosomal Ca2+ release and subsequent CaM activation. PMID:26101220

  7. [Study of antioxidant and membrane activity of rosmarinic acid using different model systems].

    PubMed

    Popov, A M; Osipov, A N; Korepanova, E A; Krivoshapko, O N; Artiukov, A A

    2013-01-01

    Rosmarinic acid is found in many species of different families of higher plants and its chemical structure is phenol propanoid with various biological activity. In this paper, we conducted a comparative study of antioxidant (radical-scavenging) properties of rosmarinic acid in systems of 2,2'-azo-bis(2-methylpropionamidin)dihydrochloride-luminol and hemoglobin-hydrogen peroxide-lu- minol, determined its protective potential in preventing peroxidation of linoleic acid, and evaluated the effect on the permeability of planar bilayer lipid membranes. Linoleic acid peroxidation was assessed by iron-thiocyanate method. In these studies, trolox was used as a reference antioxidant, and ascorbic acid, and dihydroquercetin were taken as standards. Rosmarinic acid is significantly superior to trolox, ascorbic acid and dihydroquercetin in the tests for antioxidant activity in the systems studied, as well as in inhibition of linoleic acid peroxidation. According to their activity the investigated substances can be arranged in the following order: rosmarinic acid > dihydroquercetin trolox > ascorbic acid. Rosmarinic acid does not cause significant changes in the permeability of planar bilayer membranes in a dose range of 0.5 to 10 mkg/mL. Antioxidant activity of rosmarinic acid is due to the neutralization of reactive oxygen species and/or luminol radicals generated in model systems. The observed features of the antioxidant and membrane activity of rosmarinic acid, which may underlie the previously mentioned pharmacological effects are discussed. PMID:25481945

  8. [Study of antioxidant and membrane activity of rosmarinic acid using different model systems].

    PubMed

    2013-01-01

    Rosmarinic acid is found in many species of different families of higher plants and its chemical structure is phenol propanoid with various biological activity. In this paper, we conducted a comparative study of antioxidant (radical-scavenging) properties of rosmarinic acid in systems of 2,2'-azo-bis(2-methylpropionamidin)dihydrochloride-luminol and hemoglobin-hydrogen peroxide-lu- minol, determined its protective potential in preventing peroxidation of linoleic acid, and evaluated the effect on the permeability of planar bilayer lipid membranes. Linoleic acid peroxidation was assessed by iron-thiocyanate method. In these studies, trolox was used as a reference antioxidant, and ascorbic acid, and dihydroquercetin were taken as standards. Rosmarinic acid is significantly superior to trolox, ascorbic acid and dihydroquercetin in the tests for antioxidant activity in the systems studied, as well as in inhibition of linoleic acid peroxidation. According to their activity the investigated substances can be arranged in the following order: rosmarinic acid > dihydroquercetin trolox > ascorbic acid. Rosmarinic acid does not cause significant changes in the permeability of planar bilayer membranes in a dose range of 0.5 to 10 mkg/mL. Antioxidant activity of rosmarinic acid is due to the neutralization of reactive oxygen species and/or luminol radicals generated in model systems. The observed features of the antioxidant and membrane activity of rosmarinic acid, which may underlie the previously mentioned pharmacological effects are discussed. PMID:25508797

  9. Membrane-Targeting DCAP Analogues with Broad-Spectrum Antibiotic Activity against Pathogenic Bacteria.

    PubMed

    Hurley, Katherine A; Heinrich, Victoria A; Hershfield, Jeremy R; Demons, Samandra T; Weibel, Douglas B

    2015-04-01

    We performed a structure-activity relationship study of 2-((3-(3,6-dichloro-9H-carbazol-9-yl)-2-hydroxypropyl)amino)-2-(hydroxymethyl)propane-1,3-diol (DCAP), which is an antibacterial agent that disrupts the membrane potential and permeability of bacteria. The stereochemistry of DCAP had no effect on the biological activity of DCAP. The aromaticity and electronegativity of the chlorine-substituted carbazole was required for activity, suggesting that its planar and dipolar characteristics orient DCAP in membranes. Increasing the hydrophobicity of the tail region of DCAP enhanced its antibiotic activity. Two DCAP analogues displayed promising antibacterial activity against the BSL-3 pathogens Bacillus anthracis and Francisella tularensis. Codosing DCAP analogues with ampicillin or kanamycin increased their potency. These studies demonstrate that DCAP and its analogues may be a promising scaffold for developing chemotherapeutic agents that bind to bacterial membranes and kill strains of slow-growing or dormant bacteria that cause persistent infections. PMID:25941556

  10. Membrane-Targeting DCAP Analogues with Broad-Spectrum Antibiotic Activity against Pathogenic Bacteria

    PubMed Central

    2015-01-01

    We performed a structure–activity relationship study of 2-((3-(3,6-dichloro-9H-carbazol-9-yl)-2-hydroxypropyl)amino)-2-(hydroxymethyl)propane-1,3-diol (DCAP), which is an antibacterial agent that disrupts the membrane potential and permeability of bacteria. The stereochemistry of DCAP had no effect on the biological activity of DCAP. The aromaticity and electronegativity of the chlorine-substituted carbazole was required for activity, suggesting that its planar and dipolar characteristics orient DCAP in membranes. Increasing the hydrophobicity of the tail region of DCAP enhanced its antibiotic activity. Two DCAP analogues displayed promising antibacterial activity against the BSL-3 pathogens Bacillus anthracis and Francisella tularensis. Codosing DCAP analogues with ampicillin or kanamycin increased their potency. These studies demonstrate that DCAP and its analogues may be a promising scaffold for developing chemotherapeutic agents that bind to bacterial membranes and kill strains of slow-growing or dormant bacteria that cause persistent infections. PMID:25941556

  11. Methanogenic activities in anaerobic membrane bioreactors (AnMBR) treating synthetic municipal wastewater.

    PubMed

    Ho, Jaeho; Sung, Shihwu

    2010-04-01

    Two laboratory-scale anaerobic membrane bioreactors, AnMBR 1 and AnMBR 2, were run in parallel at 25 and 15 degrees C, respectively. Total chemical oxygen demand (COD) removal efficiency was more than 95% and 85% for AnMBR 1 and 2, respectively. The COD removal of AnMBR 1 was mostly carried out biologically. However, the physical removal on the membrane surface compensated for the decreased biological removal rate in AnMBR 2. The membrane in AnMBR systems is likely not only to retain all biomass in the reactor, but also complement decreased biological removal efficiency at low temperature by rejecting soluble organics. Specific methanogenic activity (SMA) test was used to investigate the methanogenic activity profiles of suspended and attached sludge in AnMBRs treating synthetic municipal wastewater at 25 and 15 degrees C. The methanogenic activity was 51.8 ml CH(4)/g VSSd on day 1 and eventually increased 27% and reached 65.7 ml CH(4)/g VSSd on day 75 for AnMBR 1. However, the methanogenic activity of AnMBR 2 sludge was lower than that of AnMBR 1. The microbial activity of suspended sludge continuously increased, while that of attached sludge gradually decreased in this study. The methanogenic activity of attached sludge was far lower than that of suspended sludge. The role of attached sludge on the membrane in AnMBRs as a biofilm for biological organic removal was minimal compared to suspended sludge. PMID:20022745

  12. Arrayed lipid bilayer chambers allow single-molecule analysis of membrane transporter activity

    PubMed Central

    Watanabe, Rikiya; Soga, Naoki; Fujita, Daishi; Tabata, Kazuhito V.; Yamauchi, Lisa; Hyeon Kim, Soo; Asanuma, Daisuke; Kamiya, Mako; Urano, Yasuteru; Suga, Hiroaki; Noji, Hiroyuki

    2014-01-01

    Nano- to micron-size reaction chamber arrays (femtolitre chamber arrays) have facilitated the development of sensitive and quantitative biological assays, such as single-molecule enzymatic assays, digital PCR and digital ELISA. However, the versatility of femtolitre chamber arrays is limited to reactions that occur in aqueous solutions. Here we report an arrayed lipid bilayer chamber system (ALBiC) that contains sub-million femtolitre chambers, each sealed with a stable 4-μm-diameter lipid bilayer membrane. When reconstituted with a limiting amount of the membrane transporter proteins α-hemolysin or F0F1-ATP synthase, the chambers within the ALBiC exhibit stochastic and quantized transporting activities. This demonstrates that the single-molecule analysis of passive and active membrane transport is achievable with the ALBiC system. This new platform broadens the versatility of femtolitre chamber arrays and paves the way for novel applications aimed at furthering our mechanistic understanding of membrane proteins’ function. PMID:25058452

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

    SciTech Connect

    Kinsella, J.; Sacktor, B.

    1985-06-01

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

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

    PubMed

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

    2014-03-01

    Photodynamic tumor-destroying activity of the boronated chlorin e6 derivative BACE (chlorin e6 13(1)-N-{2-[N-(1-carba-closo-dodecaboran-1-yl)methyl]aminoethyl}amide-15(2), 17(3)-dimethyl ester), previously described in Moisenovich et al. (2010) PLoS ONE 5(9) e12717, was shown here to be enormously higher than that of unsubstituted chlorin e6, being supported by the data on much higher photocytotoxicity of BACE in M-1 sarcoma cell culture. To validate membrane damaging effect as the basis of the enhanced tumoricidal activity, BACE was compared with unsubstituted chlorin e6 in the potency to photosensitize dye leakage from liposomes, transbilayer lipid flip-flop, inactivation of gramicidin A ionic channels in planar lipid membranes and erythrocyte hemolysis. In all the models comprising artificial and cellular membranes, the photodynamic effect of BACE exceeded that of chlorin e6. BACE substantially differed from chlorin e6 in the affinity to liposomes and erythrocytes, as monitored by fluorescence spectroscopy, flow cytometry and centrifugation. The results support the key role of membrane binding in the photodynamic effect of the boronated chlorin e6 amide. PMID:24287152

  15. pH regulation of amphotericin B channels activity in the bilayer lipid membrane

    PubMed Central

    Shahmoradi, Tahereh; Sepehry, Hamid; Ashrafpour, Manuchehr

    2016-01-01

    Background: Amphotericin B (AmB) is a polyene antibiotic frequently applied in the treatment of systemic fungal infections in spite of its secondary effects. The pH plays a crucial role in modulating biophysical features of ion channels in the bilayer lipid membranes. Aim: In this study, the role of pH in the regulation of AmB channel was assessed by single channel recording of ion channel incorporated in the artificial membrane. Materials and Methods: Bilayer lipid membrane was formed by phosphatidylcholine in a 350 μm diameter aperture between two chambers, cis and trans contained 200/50 mMKCl solutions, respectively; then AmB was incorporated into the bilayer lipid membrane. Single channel recordings were used to indicate the effects of pH changes on AmB channels activity. The records were analyzed by Clamp fit 10 software. Results: A kinetic analysis of single channel currents indicated a cation ion channel with 500 pS conductance and voltage-dependence of the open probability of the AmB channel (Po). A reduction of cis pH to 6 decreased Po and conductance. This effect was also voltage-dependent, being greater at a more positive above −40. The pH changes in the range of 6-8 had no effect on the reversal potential and ion selectivity. Conclusion: Our data indicated that extracellular acidity can reduce AmB activity. PMID:27003977

  16. Comparative Transport Activity of Intact Cells, Membrane Vesicles, and Mesosomes of Bacillus licheniformis

    PubMed Central

    MacLeod, Robert A.; Thurman, Paul; Rogers, H. J.

    1973-01-01

    Sodium ion was shown to stimulate strongly the transport of l-glutamic acid into cells of Bacillus licheniformis 6346 His−. Lithium ion had a slight capacity to replace Na+ in this capacity, but K+ was without effect. Three of five amino acids tested. l-glutamic acid, l-aspartic acid, and l-alanine, were concentrated against a gradient in the cells. Intracellular pools of these amino acids were extractable with 5% trichloroacetic acid. Pools of l-histidine and l-lysine could not be detected. No evidence of active transport of lysine into cells could be detected, and histidine was taken up in the absence of chloramphenicol but not in its presence. The uptake of glutamic acid by membrane vesicle preparations was strongly stimulated by reduced nicotinamide adenine dinucleotide (NADH) and to a lesser extent by succinate. The presence of phenazine methosulfate increased uptake in the presence of succinate. Either l- or d-lactate and adenosine triphosphate were without effect. None of these compounds stimulated the uptake of glutamic acid by mesosomes, although some mesosome preparations contained separable membrane which was very active. NADH strongly stimulated the uptake of aspartic acid and alanine by membrane vesicles but had only a slight effect on the uptake of histidine and lysine. No evidence of active transport of any of the amino acids into mesosomes could be detected either in the presence or absence of NADH. NADH stimulation of the uptake of glutamic acid by membrane vesicles was destroyed by exposure to light of 360 nm; this inactivation was reversible by vitamin K2(5) or K2(10). Sodium ion stimulated transport of glutamic acid by membrane vesicles. PMID:4347247

  17. Infrared Laser Activation of Soluble and Membrane Protein Assemblies in the Gas Phase.

    PubMed

    Mikhailov, Victor A; Liko, Idlir; Mize, Todd H; Bush, Matthew F; Benesch, Justin L P; Robinson, Carol V

    2016-07-19

    Collision-induced dissociation (CID) is the dominant method for probing intact macromolecular complexes in the gas phase by means of mass spectrometry (MS). The energy obtained from collisional activation is dependent on the charge state of the ion and the pressures and potentials within the instrument: these factors limit CID capability. Activation by infrared (IR) laser radiation offers an attractive alternative as the radiation energy absorbed by the ions is charge-state-independent and the intensity and time scale of activation is controlled by a laser source external to the mass spectrometer. Here we implement and apply IR activation, in different irradiation regimes, to study both soluble and membrane protein assemblies. We show that IR activation using high-intensity pulsed lasers is faster than collisional and radiative cooling and requires much lower energy than continuous IR irradiation. We demonstrate that IR activation is an effective means for studying membrane protein assemblies, and liberate an intact V-type ATPase complex from detergent micelles, a result that cannot be achieved by means of CID using standard collision energies. Notably, we find that IR activation can be sufficiently soft to retain specific lipids bound to the complex. We further demonstrate that, by applying a combination of collisional activation, mass selection, and IR activation of the liberated complex, we can elucidate subunit stoichiometry and the masses of specifically bound lipids in a single MS experiment. PMID:27328020

  18. Preparation of the superhydrophobic nano-hybrid membrane containing carbon nanotube based on chitosan and its antibacterial activity.

    PubMed

    Song, Kaili; Gao, Aiqin; Cheng, Xi; Xie, Kongliang

    2015-10-01

    The functional nano-hybrid surface containing multi-walled carbon nanotubes (MWCNT) on chitosan incorporated with the cationic chitosan (C-CS), MWCNTs and silicon couple agent (KH-560) was designed and prepared. The nano-hybrid membranes (NHM) containing MWCNTs were modified by perfluorooctanesulfonyl fluoride (PFOSF). The superhydrophobic multi-functional membranes with biological activity and superhydrophobic surface were obtained. The incorporated MWCNTs improved the roughness of the nano-hybrid membranes. The perfluorinated end groups of the nano-hybrid membrane surface provided low energy surface. The antibacterial activity, surface superhydrophobicity and mechanical property of the perfluorinated nano-hybrid membranes (PFNM) were discussed. Their morphological structures and surface ingredients were characterized by energy dispersive X-ray spectrometer (SEM-EDX). The PFNMs had excellent antibacterial property and superhydrophobicity. The novel nano-hybrid membranes with excellent antibacterial, superhydrophbic, and mechanical properties have potential applications in the food engineering, bioengineering fields and medical materials. PMID:26076639

  19. Cationic Membrane Peptides: Atomic-Level Insight of Structure-Activity Relationships from Solid-State NMR

    PubMed Central

    Su, Yongchao; Li, Shenhui; Hong, Mei

    2012-01-01

    Many membrane-active peptides, such as cationic cell-penetrating peptides (CPPs) and antimicrobial peptides (AMPs), conduct their biological functions by interacting with the cell membrane. The interactions of charged residues with lipids and water facilitate membrane insertion, translocation or disruption of these highly hydrophobic species. In this mini-review we will summarize high-resolution structural and dynamic findings towards the understanding of the structure-activity relationship of lipid membrane-bound CPPs and AMPs, as examples of the current development of solid-state NMR (SSNMR) techniques for studying membrane peptides. We will present the most recent atomic-resolution structure of the guanidinium-phosphate complex, as constrained from experimentally measured site-specific distances. These SSNMR results will be valuable specifically for understanding the intracellular translocation pathway of CPPs and antimicrobial mechanism of AMPs, and more generally broaden our insight into how cationic macromolecules interact with and cross the lipid membrane. PMID:23108593

  20. Activity of key enzymes in microsomal and mitochondrial membranes depends on the redox reactions involving lipid radicals.

    PubMed

    Dmitriev, L F

    2001-07-01

    The work reviews membrane processes, such as monooxygenase reaction and oxidative phosphorylation with special reference to hydroxylation of a xenobiotic benzo(a)pyrene and the effects of the radical scavenger propyl gallate and radical generator Fe2+ ions on the reaction kinetics. A possibility is discussed that tocopherol provides for the activity of the lipid-radical cycles involving cytochrome b5. The lipid-radical cycles protect membrane lipids from oxidation and control the kinetics of membrane processes. The NADPH oxidation energy is transformed into the energy of lipid pulsations and this energy is used for activation of membrane enzymes. To account for the role of lipid pulsations in membrane processes, a new parameter is introduced - the internal temperature. It is supposed that there should be the equilibrium between the pro- and antioxidant factors in the membranes, and the presence of exogenous antioxidants (propyl gallate etc.) should be considered as a negative factor. PMID:11699868

  1. The plasma membrane shuttling of CAPRI is related to regulation of mast cell activation

    SciTech Connect

    Nakamura, Rika; Furuno, Tadahide; Nakanishi, Mamoru . E-mail: mamoru@dpc.agu.ac.jp

    2006-08-18

    The Ca{sup 2+}-promoted Ras inactivator (CAPRI), a Ras GTPase-activating protein, is involved in the inactivation of mitogen-activated protein kinase pathway. However, a precise role of CAPRI in immune responses is still unknown. Here we showed that overexpression of CAPRI suppresses antigen-induced degranulation and cytokine production in mast cells (RBL cells). Antigen elicited the translocation of CAPRI to the plasma membrane from the cytoplasm, which was concomitant with the increase in the intracellular Ca{sup 2+} concentration. The nuclear import of extracellular signal-regulated kinase 2 (ERK2) occurred after the re-localization of CAPRI to the cytoplasm in the mast cells, suggesting that the early phase of ERK2 activation is eliminated. A mutant of GAP-related domain, CAPRI(R472S), showed a feeble translocation to the plasma membrane but did not affect the degranulation, ERK2 activation, and cytokine production. The results suggested that the translocation of CAPRI to the plasma membranes regulates crucially cellular responses in mast cells.

  2. Plasma Membrane Intrinsic Proteins from Maize Cluster in Two Sequence Subgroups with Differential Aquaporin Activity1

    PubMed Central

    Chaumont, François; Barrieu, François; Jung, Rudolf; Chrispeels, Maarten J.

    2000-01-01

    The transport of water through membranes is regulated in part by aquaporins or water channel proteins. These proteins are members of the larger family of major intrinsic proteins (MIPs). Plant aquaporins are categorized as either tonoplast intrinsic proteins (TIPs) or plasma membrane intrinsic proteins (PIPs). Sequence analysis shows that PIPs form several subclasses. We report on the characterization of three maize (Zea mays) PIPs belonging to the PIP1 and PIP2 subfamilies (ZmPIP1a, ZmPIP1b, and ZmPIP2a). The ZmPIP2a clone has normal aquaporin activity in Xenopus laevis oocytes. ZmPIP1a and ZmPIP1b have no activity, and a review of the literature shows that most PIP1 proteins identified in other plants have no or very low activity in oocytes. Arabidopsis PIP1 proteins are the only exception. Control experiments show that this lack of activity of maize PIP1 proteins is not caused by their failure to arrive at the plasma membrane of the oocytes. ZmPIP1b also does not appear to facilitate the transport of any of the small solutes tried (glycerol, choline, ethanol, urea, and amino acids). These results are discussed in relationship to the function and regulation of the PIP family of aquaporins. PMID:10759498

  3. Ethylene activates a plasma membrane Ca(2+)-permeable channel in tobacco suspension cells.

    PubMed

    Zhao, Min-Gui; Tian, Qiu-Ying; Zhang, Wen-Hao

    2007-01-01

    Here, the effects of the ethylene-releasing compound, ethephon, and the ethylene precursor, 1-aminocyclopropane-1-carboxylic acid (ACC), on ionic currents across plasma membranes and on the cytosolic Ca(2+) activity ([Ca(2+)](c)) of tobacco (Nicotiana tabacum) suspension cells were characterized using a patch-clamp technique and confocal laser scanning microscopy. Exposure of tobacco protoplasts to ethephon and ACC led to activation of a plasma membrane cation channel that was permeable to Ba(2+), Mg(2+) and Ca(2+), and inhibited by La(3+), Gd(3+) and Al(3+). The ethephon- and ACC-induced Ca(2+)-permeable channel was abolished by the antagonist of ethylene perception (1-metycyclopropene) and by the inhibitor of ACC synthase (aminovinylglycin), indicating that activation of the Ca(2+)-permeable channels results from ethylene. Ethephon elicited an increase in the [Ca(2+)](c) of tobacco suspension cells, as visualized by the Ca(2+)-sensitive probe Fluo-3 and confocal microscopy. The ethephon-induced elevation of [Ca(2+)](c) was markedly inhibited by Gd(3+) and BAPTA, suggesting that an influx of Ca(2+) underlies the elevation of [Ca(2+)](c). These results indicate that an elevation of [Ca(2+)](c), resulting from activation of the plasma membrane Ca(2+)-permeable channels by ethylene, is an essential component in ethylene signaling in plants. PMID:17447907

  4. Impact of humic acid fouling on membrane performance and transport of pharmaceutically active compounds in forward osmosis.

    PubMed

    Xie, Ming; Nghiem, Long D; Price, William E; Elimelech, Menachem

    2013-09-01

    The impact of humic acid fouling on the membrane transport of two pharmaceutically active compounds (PhACs) - namely carbamazepine and sulfamethoxazole - in forward osmosis (FO) was investigated. Deposition of humic acid onto the membrane surface was promoted by the complexation with calcium ions in the feed solution and the increase in ionic strength at the membrane surface due to the reverse transport of NaCl draw solute. The increase in the humic acid deposition on the membrane surface led to a substantial decrease in the membrane salt (NaCl) permeability coefficient but did not result in a significant decrease in the membrane pure water permeability coefficient. As the deposition of humic acid increased, the permeation of carbamazepine and sulfamethoxazole decreased, which correlated well with the decrease in the membrane salt (NaCl) permeability coefficient. It is hypothesized that the hydrated humic acid fouling layer hindered solute diffusion through the membrane pore and enhanced solute rejection by steric hindrance, but not the permeation of water molecules. The membrane water and salt (NaCl) permeability coefficients were fully restored by physical cleaning of the membrane, suggesting that humic acid did not penetrate into the membrane pores. PMID:23764606

  5. 5-Lipoxygenase-activating protein rescues activity of 5-lipoxygenase mutations that delay nuclear membrane association and disrupt product formation.

    PubMed

    Gerstmeier, Jana; Newcomer, Marcia E; Dennhardt, Sophie; Romp, Erik; Fischer, Jana; Werz, Oliver; Garscha, Ulrike

    2016-05-01

    Leukotrienes (LTs) are proinflammatory lipid mediators formed from arachidonic acid in a 2-step reaction catalyzed by 5-lipoxygenase (5-LOX) requiring the formation of 5-HPETE [5(S)-hydroperoxy-6-trans-8,11,14-cis-eicosatetraenoic acid] and its subsequent transformation to LTA4 5-LOX is thought to receive arachidonic acid from the nuclear membrane-embedded 5-LOX-activating protein (FLAP). The crystal structure of 5-LOX revealed an active site concealed by F177 and Y181 (FY cork). We examined the influence of the FY cork on 5-LOX activity and membrane binding in HEK293 cells in the absence and presence of FLAP. Uncapping the 5-LOX active site by mutation of F177 and/or Y181 to alanine (5-LOX-F177A, 5-LOX-Y181A, 5-LOX-F177/Y181A) resulted in delayed and diminished 5-LOX membrane association in A23187-stimulated cells. For 5-LOX-F177A and 5-LOX-F177/Y181A, formation of 5-LOX products was dramatically reduced relative to 5-LOX-wild type (wt). Strikingly, coexpression of FLAP in A23187-activated HEK293 cells effectively restored formation of 5-H(p)ETE (5-hydroxy- and 5-peroxy-6-trans-8,11,14-cis-eicosatetraenoic acid) by these same 5-LOX mutants (≈60-70% 5-LOX-wt levels) but not of LTA4 hydrolysis products. Yet 5-LOX-Y181A generated 5-H(p)ETE at levels comparable to 5-LOX-wt but reduced LTA4 hydrolysis products. Coexpression of FLAP partially restored LTA4 hydrolysis product formation by 5-LOX-Y181A. Together, the data suggest that the concealed FY cork impacts membrane association and that FLAP may help shield an uncapped active site.-Gerstmeier, J., Newcomer, M. E., Dennhardt, S., Romp, E., Fischer, J., Werz, O., Garscha, U. 5-Lipoxygenase-activating protein rescues activity of 5-lipoxygenase mutations that delay nuclear membrane association and disrupt product formation. PMID:26842853

  6. Ethacrynic acid inhibitable Ca2+ and Mg2+-activated membrane adenosine triphosphatase in rat mast cells.

    PubMed Central

    Magro, A M

    1977-01-01

    A crude plasma membrane fraction from the homogenate of purified rat mast cells demonstrates a high degree of Ca2+-dependent and Mg2+-dependent adenosine triphosphatase (ATPase) activity. The microsomal and mitochondrial fractions show negligible amounts of the Ca2+ and Mg2+-activated ATPases. The broad ATPase inhibitor, ethacrynic acid, effectively blocks the mast cell ATPase activity while ouabain demonstrates little inhibitory effect. Correspondingly, ethacrynic acid inhibits histamine release from antigen-challenged mast cells while ouabain does not. Both ATPase inhibition and histamine release inhibition by ethacrynic acid require the presence of the olefinic bond in the ethacrynic acid molecule. PMID:75076

  7. [X-ray microanalysis of the activity of immobilized urease on chitosan membrane].

    PubMed

    Ma, Xiao-li; Yao, Zi-hua

    2005-03-01

    The localization of the activity of immobilized urease on chitosan membrane was studied by X-ray microanalysis. BaCl2 and urea were selected as the capture and substrate respectively. The substrate was hydrolyzed by immobilized urease to produce NH3 and CO2 in Tris-HCl buffer (pH 7.0), and the latter was captured by BaCl2 to form precipitate. The precipite was deposited on the active site of immobilized urease. It is shown that the method is practicable and reliable. The optimum condition for the localization of activity of immobilized urease was studied. PMID:16013332

  8. Effect of serum lipoproteins on the adenylate cyclase activity of rat liver plasma membranes.

    PubMed Central

    Ghiselli, G; Sirtori, C R; Nicosia, S

    1981-01-01

    Four rat lipoprotein classes [lymph chylomicrons, VLD (very-low-density), LD (low-density) and HD (high-density) lipoproteins] were tested for their ability to affect basal adenylate cyclase (EC 4.6.1.1) activity of rat liver plasma membranes. All the lipoproteins, with the exception of lymph chylomicrons, effectively increase the enzyme activity. VLD lipoproteins are the most active class (67% maximal increase), followed by HD lipoproteins (33%) and LD lipoproteins (23%). The effect of VLD lipoproteins is additive to that elicited by GTP or GTP plus glucagon (at least within a certain concentration range). VLD lipoproteins affect only the Vmax. of the enzyme, not the Km. PMID:7317023

  9. Gαi3-Dependent Inhibition of JNK Activity on Intracellular Membranes

    PubMed Central

    Bastin, Guillaume; Yang, Jin Ye; Heximer, Scott P.

    2015-01-01

    Heterotrimeric G-protein signaling has been shown to modulate a wide variety of intracellular signaling pathways, including the mitogen-activated protein kinase (MAPK) family. The activity of one MAPK family class, c-Jun N-terminal kinases (JNKs), has been traditionally linked to the activation of G-protein coupled receptors (GPCRs) at the plasma membrane. Using a unique set of G-protein signaling tools developed in our laboratory, we show that subcellular domain-specific JNK activity is inhibited by the activation of Gαi3, the Gαi isoform found predominantly within intracellular membranes, such as the endoplasmic reticulum (ER)–Golgi interface, and their associated vesicle pools. Regulators of intracellular Gαi3, including activator of G-protein signaling 3 (AGS3) and the regulator of G-protein signaling protein 4 (RGS4), have a marked impact on the regulation of JNK activity. Together, these data support the existence of unique intracellular signaling complexes that control JNK activity deep within the cell. This work highlights some of the cellular pathways that are regulated by these intracellular complexes and identifies potential strategies for their regulation in mammalian cells. PMID:26389115

  10. Membrane disruptive antimicrobial activities of human β-defensin-3 analogs.

    PubMed

    Sudheendra, U S; Dhople, Vishnu; Datta, Aritreyee; Kar, Rajiv K; Shelburne, Charles E; Bhunia, Anirban; Ramamoorthy, Ayyalusamy

    2015-02-16

    Human beta defensin-3 (HβD-3) is a host-defense protein exhibiting antibacterial activity towards both Gram-negative and Gram-positive bacteria. There is considerable interest in the function of this protein due to its increased salt tolerance and activity against Gram-positive Staphylococcus aureus. In this study, analogs of HβD-3 devoid of N and C terminal regions are investigated to determine the influence of specific structural motif on antimicrobial activity and selectivity between Gram-positive and Gram-negative bacteria. Circular dichroism, fluorescence and solid-state NMR experiments have been used to investigate the conformation and mode of action of HβD3 analogs with various model membranes to mimic bacterial inner and outer membranes and also mammalian membranes. Our studies specifically focused on determining four major characteristics: (i) interaction of HβD3 analogs with phospholipid vesicles composed of zwitterionic PC or anionic PE:PG vesicles and LPS; (ii) conformation of HβD3-peptide analogs in the presence of PC or PE:PG vesicles; (iii) ability of HβD3 analogs to permeate phospholipid vesicles composed of PC or PE:PG; and (iv) activities on bacteria cells and erythrocytes. Our results infer that the linear peptide L25P and its cyclic form C25P are more active than L21P and C21P analogs. However, they are less active than the parent peptide, thus pointing towards the importance of the N terminal domain in its biological activity. The variation in the activities of L21P/C21P and L25P/C25P also suggest the importance of the positively charged residues at the C terminus in providing selectivity particularly to Gram-negative bacteria. PMID:25112689

  11. Membrane Disruptive Antimicrobial Activities of Human β-Defensin-3 Analogs

    PubMed Central

    Sudheendra, U. S.; Dhople, Vishnu; Datta, Aritreyee; Kar, Rajiv K.; Shelburne, Charles E.; Bhunia, Anirban; Ramamoorthy, Ayyalusamy

    2014-01-01

    Human beta defensin-3 (HβD-3) is a host-defense protein exhibiting antibacterial activity towards both Gram-negative and Gram-positive bacteria. There is considerable interest in the function of this protein due to its increased salt tolerance and activity against Gram-positive S. aureus. In this study, analogs of HβD-3 devoid of N and C terminal regions are investigated to determine the influence of specific structural motif on antimicrobial activity and selectivity between Gram-positive and Gram-negative bacteria. Circular dichroism, fluorescence and solid-state NMR experiments have been used to investigate the conformation and mode of action of HβD3 analogs with various model membranes to mimic bacterial inner and outer membranes and also mammalian membranes. Our studies specifically focused on determining four major characteristics: (i) interaction of HβD3 analogs with phospholipid vesicles composed of zwitterionic PC or anionic PE:PG vesicles and LPS; (ii) conformation of HβD3-peptide analogs in the presence of PC or PE:PG vesicles; (iii) ability of HβD3 analogs to permeate phospholipid vesicles composed of PC or PE:PG; and (iv) activities on bacteria cells and erythrocytes. Our results infer that the linear peptide L25P and its cyclic form C25P are more active than L21P and C21P analogs. However, they are less active than the parent peptide, thus pointing towards the importance of the N terminal domain in its biological activity. The variation in the activities of L21P/C21P and L25P/C25P also suggest the importance of the positively charged residues at the C terminus in providing selectivity particularly to Gram-negative bacteria. PMID:25112689

  12. Beyond apoptosis: the mechanism and function of phosphatidylserine asymmetry in the membrane of activating mast cells.

    PubMed

    Rysavy, Noel M; Shimoda, Lori M N; Dixon, Alyssa M; Speck, Mark; Stokes, Alexander J; Turner, Helen; Umemoto, Eric Y

    2014-01-01

    Loss of plasma membrane asymmetry is a hallmark of apoptosis, but lipid bilayer asymmetry and loss of asymmetry can contribute to numerous cellular functions and responses that are independent of programmed cell death. Exofacial exposure of phosphatidylserine occurs in lymphocytes and mast cells after antigenic stimulation and in the absence of apoptosis, suggesting that there is a functional requirement for phosphatidylserine exposure in immunocytes. In this review we examine current ideas as to the nature of this functional role in mast cell activation. Mechanistically, there is controversy as to the candidate proteins responsible for phosphatidylserine translocation from the internal to external leaflet, and here we review the candidacies of mast cell PLSCR1 and TMEM16F. Finally we examine the potential relationship between functionally important mast cell membrane perturbations and phosphatidylserine exposure during activation. PMID:25759911

  13. Beyond apoptosis: The mechanism and function of phosphatidylserine asymmetry in the membrane of activating mast cells

    PubMed Central

    Rysavy, Noel M.; Shimoda, Lori M. N.; Dixon, Alyssa M.; Speck, Mark; Stokes, Alexander J.; Turner, Helen; Umemoto, Eric Y.

    2014-01-01

    Loss of plasma membrane asymmetry is a hallmark of apoptosis, but lipid bilayer asymmetry and loss of asymmetry can contribute to numerous cellular functions and responses that are independent of programmed cell death. Exofacial exposure of phosphatidylserine occurs in lymphocytes and mast cells after antigenic stimulation and in the absence of apoptosis, suggesting that there is a functional requirement for phosphatidylserine exposure in immunocytes. In this review we examine current ideas as to the nature of this functional role in mast cell activation. Mechanistically, there is controversy as to the candidate proteins responsible for phosphatidylserine translocation from the internal to external leaflet, and here we review the candidacies of mast cell PLSCR1 and TMEM16F. Finally we examine the potential relationship between functionally important mast cell membrane perturbations and phosphatidylserine exposure during activation. PMID:25759911

  14. A Rapid and Quantitative Flow Cytometry Method for the Analysis of Membrane Disruptive Antimicrobial Activity

    PubMed Central

    O’Brien-Simpson, Neil M.; Pantarat, Namfon; Attard, Troy J.; Walsh, Katrina A.; Reynolds, Eric C.

    2016-01-01

    We describe a microbial flow cytometry method that quantifies within 3 hours antimicrobial peptide (AMP) activity, termed Minimum Membrane Disruptive Concentration (MDC). Increasing peptide concentration positively correlates with the extent of bacterial membrane disruption and the calculated MDC is equivalent to its MBC. The activity of AMPs representing three different membranolytic modes of action could be determined for a range of Gram positive and negative bacteria, including the ESKAPE pathogens, E. coli and MRSA. By using the MDC50 concentration of the parent AMP, the method provides high-throughput, quantitative screening of AMP analogues. A unique feature of the MDC assay is that it directly measures peptide/bacteria interactions and lysed cell numbers rather than bacteria survival as with MIC and MBC assays. With the threat of multi-drug resistant bacteria, this high-throughput MDC assay has the potential to aid in the development of novel antimicrobials that target bacteria with improved efficacy. PMID:26986223

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

    NASA Technical Reports Server (NTRS)

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

    2003-01-01

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

  16. Low temperature alters plasma membrane lipid composition and ATPase activity of pineapple fruit during blackheart development.

    PubMed

    Zhou, Yuchan; Pan, Xiaoping; Qu, Hongxia; Underhill, Steven J R

    2014-02-01

    Plasma membrane (PM) plays central role in triggering primary responses to chilling injury and sustaining cellular homeostasis. Characterising response of membrane lipids to low temperature can provide important information for identifying early causal factors contributing to chilling injury. To this end, PM lipid composition and ATPase activity were assessed in pineapple fruit (Ananas comosus) in relation to the effect of low temperature on the development of blackheart, a form of chilling injury. Chilling temperature at 10 °C induced blackheart development in concurrence with increase in electrolyte leakage. PM ATPase activity was decreased after 1 week at low temperature, followed by a further decrease after 2 weeks. The enzyme activity was not changed during 25 °C storage. Loss of total PM phospholipids was found during postharvest senescence, but more reduction was shown from storage at 10 °C. Phosphatidylcholine and phosphatidylethanolamine were the predominant PM phospholipid species. Low temperature increased the level of phosphatidic acid but decreased the level of phosphatidylinositol. Both phospholipid species were not changed during storage at 25 °C. Postharvest storage at both temperatures decreased the levels of C18:3 and C16:1, and increased level of C18:1. Low temperature decreased the level of C18:2 and increased the level of C14:0. Exogenous application of phosphatidic acid was found to inhibit the PM ATPase activity of pineapple fruit in vitro. Modification of membrane lipid composition and its effect on the functional property of plasma membrane at low temperature were discussed in correlation with their roles in blackheart development of pineapple fruit. PMID:24390546

  17. Sensor-actuator coupled device for active tracheal tube using solid polymer electrolyte membrane

    NASA Astrophysics Data System (ADS)

    Ihara, Tadashi; Nakamura, Taro; Mukai, Toshiharu; Asaka, Kinji

    2007-04-01

    A sensor-actuator coupled device was developed using solid polymer electrolyte membrane (SPM) as an active tracheal tube for ventilator. Active tracheal tube is a novel type of tube for ventilator that removes patient's phlegm automatically upon sensing the narrowing of trachea by phlegm. This type of active tube is extremely useful in clinical settings as currently the sole measure to remove phlegm from patient's tube is to do it manually by a nurse every few hours. As SPM works both as a sensor and an actuator, an effective compact device was developed. SPM based sensor-actuator coupled device was fabricated with modified gold plating method. Prepared SPM was fixed as an array on a plastic pipe of diameter 22 mm and was connected to a ventilator circuit and driven by a ventilator with a volume control ventilation (VCV) mode. SPM was connected both to a sensing unit and an actuation unit. Generated voltage developed by the membrane with the setting of the maximum pressure from 5 cmH IIO to 20 cmH IIO was in order of several hundred μV. SPM sensor demonstrated a biphasic response to the ventilator flow. The sensor data showed nearly linearly proportional voltage development to the intra-tracheal pressure. The sensed signal was filtered and digitized with an A/D converting unit on a PC board. A real time operating program was used to detect the sensed signal that indicates the narrowing of trachea. The program then activated a driving signal to control the actuation of the membrane. The signal was sent to a D/A converting unit. The output of the D/A unit was sent to an amplifier and the galvanostat unit which drives the membrane with constant current regardless of the change in the load. It was demonstrated that the sensor-actuator unit detects the narrowing of trachea within several hundreds milli-seconds and responds by actuating the same membrane with the driving voltage of 3-4 V and driving current of several hundred milli-ampere for each membrane. SPM array

  18. Branched phospholipids render lipid vesicles more susceptible to membrane-active peptides.

    PubMed

    Mitchell, Natalie J; Seaton, Pamela; Pokorny, Antje

    2016-05-01

    Iso- and anteiso-branched lipids are abundant in the cytoplasmic membranes of bacteria. Their function is assumed to be similar to that of unsaturated lipids in other organisms - to maintain the membrane in a fluid state. However, the presence of terminally branched membrane lipids is likely to impact other membrane properties as well. For instance, lipid acyl chain structure has been shown to influence the activity of antimicrobial peptides. Moreover, the development of resistance to antimicrobial agents in Staphylococcus aureus is accompanied by a shift in the fatty acid composition toward a higher fraction of anteiso-branched lipids. Little is known about how branched lipids and the location of the branch point affect the activity of membrane-active peptides. We hypothesized that bilayers containing lipids with low phase transition temperatures would tend to exclude peptides and be less susceptible to peptide-induced perturbation than those made from higher temperature melting lipids. To test this hypothesis, we synthesized a series of asymmetric phospholipids that only differ in the type of fatty acid esterified at the sn-2 position of the lipid glycerol backbone. We tested the influence of acyl chain structure on peptide activity by measuring the kinetics of release from dye-encapsulated lipid vesicles made from these synthetic lipids. The results were compared to those obtained using vesicles made from S. aureus and Staphylococcus sciuri membrane lipid extracts. Anteiso-branched phospholipids, which melt at very low temperatures, produced lipid vesicles that were only slightly less susceptible to peptide-induced dye release than those made from the iso-branched isomer. However, liposomes made from bacterial phospholipid extracts were generally much more resistant to peptide-induced perturbation than those made from any of the synthetic lipids. The results suggest that the increase in the fraction of anteiso-branched fatty acids in antibiotic-resistant strains

  19. Model Membrane and Cell Studies of Antimicrobial Activity of Melittin Analogues.

    PubMed

    Jamasbi, Elaheh; Mularski, Anna; Separovic, Frances

    2016-01-01

    Melittin is a 26 residue peptide and the major component of bee (Apis mellifera) venom. Although melittin has both anticancer and antimicrobial properties, utilization has been limited due to its high lytic activity against eukaryotic cells. The mechanism of this lytic activity remains unclear but several mechanisms have been proposed, including pore formation or a detergent like mechanism, which result in lysis of cell membranes. Several analogues of melittin have been synthesized to further understand the role of specific residues in its antimicrobial and lytic activity. Melittin analogues that have a proline residue substituted for an alanine, lysine or cysteine have been studied with both model membrane systems and living cells. These studies have revealed that the proline residue plays a critical role in antimicrobial activity and cytotoxicity. Analogues lacking the proline residue and dimers of these analogues displayed decreased cytotoxicity and minimum inhibition concentrations. Several mutant studies have shown that, when key substitutions are made, the resultant peptides have more activity in terms of pore formation than the native melittin. Designing analogues that retain antimicrobial and anticancer activity while minimizing haemolytic activity will be a promising way to utilize melittin as a potential therapeutic agent. PMID:26139117

  20. Pretreatment with alum or powdered activated carbon reduces bacterial predation-associated irreversible fouling of membranes.

    PubMed

    Kim, Eun-Ho; Dwidar, Mohammed; Kwon, Young-Nam; Mitchell, Robert J

    2014-01-01

    This study evaluated the co-application of bacterial predation by Bdellovibrio bacteriovorus and either alum coagulation or powdered activated carbon adsorption to reduce fouling caused by Escherichia coli rich feed solutions in dead-end microfiltration tests. The flux increased when the samples were predated upon or treated with 100 ppm alum or PAC, but co-treatment with alum and predation gave the best flux results. The total membrane resistance caused by the predated sample was reduced six-fold when treated with 100 ppm PAC, from 11.8 to 1.98 × 10(11) m(-1), while irreversible fouling (Rp) was 2.7-fold lower. Treatment with 100 ppm alum reduced the total resistance 14.9-fold (11.8 to 0.79 × 10(11) m(-1)) while the Rp decreased 4.25-fold. SEM imaging confirmed this, with less obvious fouling of the membrane after the combined process. This study illustrates that the combination of bacterial predation and the subsequent removal of debris using coagulation or adsorption mitigates membrane biofouling and improves membrane performance. PMID:25410737

  1. Effective lipid-detergent system for study of membrane active peptides in fluid liposomes.

    PubMed

    Sychev, Sergei V; Sukhanov, Stanislav V; Telezhinskaya, Irina N; Ovchinnikova, Tatiana V

    2016-02-01

    The structure of peptide antibiotic gramicidin A (gA) was studied in phosphatidylcholin liposomes modified by nonionic detergent Triton X-100. First, the detergent : lipid ratio at which the saturation of lipid membrane by Triton X-100 occurs (Re (sat)), was determined by light scattering. Measurements of steady-state fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene at sublytic concentrations of detergent showed that after saturation of the membrane by Triton X-100 microviscosity of lipid bilayer is reduced by 20%. The equilibrium conformational state of gA in phosphatidylcholine liposomes at Re (sat) was studied by CD spectroscopy. It was found that the conformational state of this channel-forming peptide changed crucially when Triton X-100 induced transition to more fluid membranes. The gA single-channel measurements were made with Triton X-100 containing bilayers. Tentative assignment of the channel type and gA structures was made by correlation of CD data with conductance histograms. Lipid-detergent system with variable viscosity developed in this work can be used to study the structure and folding of other membrane-active peptides. PMID:26751806

  2. Bovine serum albumin with glycated carboxyl groups shows membrane-perturbing activities.

    PubMed

    Yang, Shin-Yi; Chen, Ying-Jung; Kao, Pei-Hsiu; Chang, Long-Sen

    2014-12-15

    The aim of the present study aimed to investigate whether glycated bovine serum albumin (BSA) showed novel activities on the lipid-water interface. Mannosylated BSA (Man-BSA) was prepared by modification of the carboxyl groups with p-aminophenyl α-d-mannopyranoside. In contrast to BSA, Man-BSA notably induced membrane permeability of egg yolk phosphatidylcholine (EYPC)/egg yolk sphingomyelin (EYSM)/cholesterol (Chol) and EYPC/EYSM vesicles. Noticeably, Man-BSA induced the fusion of EYPC/EYSM/Chol vesicles, but not of EYPC/EYSM vesicles. Although BSA and Man-BSA showed similar binding affinity for lipid vesicles, the lipid-bound conformation of Man-BSA was distinct from that of BSA. Moreover, Man-BSA adopted distinct structure upon binding with the EYPC/EYSM/Chol and EYPC/EYSM vesicles. Man-BSA could induce the fusion of EYPC/EYSM/Chol vesicles with K562 and MCF-7 cells, while Man-BSA greatly induced the leakage of Chol-depleted K562 and MCF-7 cells. The modified BSA prepared by conjugating carboxyl groups with p-aminophenyl α-d-glucopyranoside also showed membrane-perturbing activities. Collectively, our data indicate that conjugation of carboxyl groups with monosaccharide generates functional BSA with membrane-perturbing activities on the lipid-water interface. PMID:25449061

  3. Porin activity of the native and recombinant outer membrane protein Oms28 of Borrelia burgdorferi.

    PubMed Central

    Skare, J T; Champion, C I; Mirzabekov, T A; Shang, E S; Blanco, D R; Erdjument-Bromage, H; Tempst, P; Kagan, B L; Miller, J N; Lovett, M A

    1996-01-01

    The outer membrane-spanning (Oms) proteins of Borrelia burgdorferi have been visualized by freeze-fracture analysis but, until recently, not further characterized. We developed a method for the isolation of B. burgdorferi outer membrane vesicles and described porin activities with single-channel conductances of 0.6 and 12.6 nS in 1 M KCI. By using both nondenaturing isoelectric focusing gel electrophoresis and fast-performance liquid chromatography separation after detergent solubilization, we found that the 0.6-nS porin activity resided in a 28-kDa protein, designated Oms28. The oms28 gene was cloned, and its nucleotide sequence was determined. The deduced amino acid sequence of Oms28 predicted a 257-amino-acid precursor protein with a putative 24-amino-acid leader peptidase I signal sequence. Processed Oms28 yielded a mature protein with a predicted molecular mass of 25,363 Da. When overproduced in Escherichia coli, the Oms28 porin fractionated in part to the outer membrane. Sodium dodecyl sulfate-polyacrylamide gel-purified recombinant Oms28 from E. coli retained functional activity as demonstrated by an average single-channel conductance of 1.1 nS in the planar lipid bilayer assay. These findings confirmed that Oms28 is a B. burgdorferi porin, the first to be described. As such, it is potential relevance to the pathogenesis of Lyme borreliosis and to the physiology of the spirochete. PMID:8759855

  4. 3,6-O-[N-(2-Aminoethyl)-acetamide-yl]-chitosan exerts antibacterial activity by a membrane damage mechanism.

    PubMed

    Yan, Feilong; Dang, Qifeng; Liu, Chengsheng; Yan, Jingquan; Wang, Teng; Fan, Bing; Cha, Dongsu; Li, Xiaoli; Liang, Shengnan; Zhang, Zhenzhen

    2016-09-20

    A novel chitosan derivative, 3,6-O-[N-(2-aminoethyl)-acetamide-yl]-chitosan (AACS), was successfully prepared to improve water solubility and antibacterial activity of chitosan. AACS had good antibacterial activity, with minimum inhibitory concentrations of 0.25mg/mL, against Escherichia coli and Staphylococcus aureus. Cell membrane integrity, electric conductivity and NPN uptake tests showed that AACS caused quickly increasing the release of intracellular nucleic acids, the uptake of NPN, and the electric conductivity by damaging membrane integrity. On the other hand, hydrophobicity, cell viability and SDS-PAGE experiments indicated that AACS was able to reduce the surface hydrophobicity, the cell viability and the intracellular proteins through increasing membrane permeability. SEM observation further confirmed that AACS could kill bacteria via disrupting their membranes. All results above verified that AACS mainly exerted antibacterial activity by a membrane damage mechanism, and it was expected to be a new food preservative. PMID:27261735

  5. Photosynthesis Activates Plasma Membrane H+-ATPase via Sugar Accumulation1[OPEN

    PubMed Central

    Okumura, Masaki; Inoue, Shin-ichiro; Kuwata, Keiko

    2016-01-01

    Plant plasma membrane H+-ATPase acts as a primary transporter via proton pumping and regulates diverse physiological responses by controlling secondary solute transport, pH homeostasis, and membrane potential. Phosphorylation of the penultimate threonine and the subsequent binding of 14-3-3 proteins in the carboxyl terminus of the enzyme are required for H+-ATPase activation. We showed previously that photosynthesis induces phosphorylation of the penultimate threonine in the nonvascular bryophyte Marchantia polymorpha. However, (1) whether this response is conserved in vascular plants and (2) the process by which photosynthesis regulates H+-ATPase phosphorylation at the plasma membrane remain unresolved issues. Here, we report that photosynthesis induced the phosphorylation and activation of H+-ATPase in Arabidopsis (Arabidopsis thaliana) leaves via sugar accumulation. Light reversibly phosphorylated leaf H+-ATPase, and this process was inhibited by pharmacological and genetic suppression of photosynthesis. Immunohistochemical and biochemical analyses indicated that light-induced phosphorylation of H+-ATPase occurred autonomously in mesophyll cells. We also show that the phosphorylation status of H+-ATPase and photosynthetic sugar accumulation in leaves were positively correlated and that sugar treatment promoted phosphorylation. Furthermore, light-induced phosphorylation of H+-ATPase was strongly suppressed in a double mutant defective in ADP-glucose pyrophosphorylase and triose phosphate/phosphate translocator (adg1-1 tpt-2); these mutations strongly inhibited endogenous sugar accumulation. Overall, we show that photosynthesis activated H+-ATPase via sugar production in the mesophyll cells of vascular plants. Our work provides new insight into signaling from chloroplasts to the plasma membrane ion transport mechanism. PMID:27016447

  6. A pore-forming toxin requires a specific residue for its activity in membranes with particular physicochemical properties.

    PubMed

    Morante, Koldo; Caaveiro, Jose M M; Tanaka, Koji; González-Mañas, Juan Manuel; Tsumoto, Kouhei

    2015-04-24

    The physicochemical landscape of the bilayer modulates membrane protein function. Actinoporins are a family of potent hemolytic proteins from sea anemones acting at the membrane level. This family of cytolysins preferentially binds to target membranes containing sphingomyelin, where they form lytic pores giving rise to cell death. Although the cytolytic activity of the actinoporin fragaceatoxin C (FraC) is sensitive to vesicles made of various lipid compositions, it is far from clear how this toxin adjusts its mechanism of action to a broad range of physiochemical landscapes. Herein, we show that the conserved residue Phe-16 of FraC is critical for pore formation in cholesterol-rich membranes such as those of red blood cells. The interaction of a panel of muteins of Phe-16 with model membranes composed of raft-like lipid domains is inactivated in cholesterol-rich membranes but not in cholesterol-depleted membranes. These results indicate that actinoporins recognize different membrane environments, resulting in a wider repertoire of susceptible target membranes (and preys) for sea anemones. In addition, this study has unveiled promising candidates for the development of protein-based biosensors highly sensitive to the concentration of cholesterol within the membrane. PMID:25759390

  7. A Pore-Forming Toxin Requires a Specific Residue for Its Activity in Membranes with Particular Physicochemical Properties*

    PubMed Central

    Morante, Koldo; Caaveiro, Jose M. M.; Tanaka, Koji; González-Mañas, Juan Manuel; Tsumoto, Kouhei

    2015-01-01

    The physicochemical landscape of the bilayer modulates membrane protein function. Actinoporins are a family of potent hemolytic proteins from sea anemones acting at the membrane level. This family of cytolysins preferentially binds to target membranes containing sphingomyelin, where they form lytic pores giving rise to cell death. Although the cytolytic activity of the actinoporin fragaceatoxin C (FraC) is sensitive to vesicles made of various lipid compositions, it is far from clear how this toxin adjusts its mechanism of action to a broad range of physiochemical landscapes. Herein, we show that the conserved residue Phe-16 of FraC is critical for pore formation in cholesterol-rich membranes such as those of red blood cells. The interaction of a panel of muteins of Phe-16 with model membranes composed of raft-like lipid domains is inactivated in cholesterol-rich membranes but not in cholesterol-depleted membranes. These results indicate that actinoporins recognize different membrane environments, resulting in a wider repertoire of susceptible target membranes (and preys) for sea anemones. In addition, this study has unveiled promising candidates for the development of protein-based biosensors highly sensitive to the concentration of cholesterol within the membrane. PMID:25759390

  8. Regulation of membrane-type 1 matrix metalloproteinase activity by vacuolar H+-ATPases.

    PubMed Central

    Maquoi, Erik; Peyrollier, Karine; Noël, Agnès; Foidart, Jean-Michel; Frankenne, Francis

    2003-01-01

    Membrane-type 1 matrix metalloproteinase (MT1-MMP) is a key enzyme in normal development and malignant processes. The regulation of MT1-MMP activity on the cell surface is a complex process involving autocatalytic processing, tissue inhibitor of MMPs (TIMP) binding and constitutive internalization. However, the fate of internalized MT1-MMP is not known. Acidification of intracellular vacuolar compartments is essential for membrane trafficking, protein sorting and degradation. This acidification is controlled by vacuolar H(+)-ATPases, which can be selectively inhibited by bafilomycin-A(1). Here, we treated human tumour cell lines expressing MT1-MMP with bafilomycin-A(1), and analysed its effects on MT1-MMP activity, internalization and processing. We show that the activity of MT1-MMP on the cell surface is constitutively down-regulated through a vacuolar H(+)-ATPase-dependent degradation process. Blockade of this degradation caused the accumulation of TIMP-free active MT1-MMP molecules on the cell surface, although internalization was not affected. As a consequence of this impaired degradation, pro-MMP-2 activation was strongly enhanced. This study demonstrates that the catalytic activity of MT1-MMP on the cell surface is regulated through a vacuolar H(+)-ATPase-dependent degradation process. PMID:12667140

  9. Mg2+ is an essential activator of hydrolytic activity of membrane-bound pyrophosphatase of Rhodospirillum rubrum.

    PubMed Central

    Sosa, A; Ordaz, H; Romero, I; Celis, H

    1992-01-01

    The substrate for the hydrolytic activity of membrane-bound pyrophosphatase is the PP(i)-Mg2+ complex. The enzyme has no activity when the free Mg2+ concentration is lower than 10 microM (at 0.5 mM-PP(i)-Mg2+), and therefore free Mg2+ is an essential activator of the hydrolytic activity. The Km for the substrate changes in response to variation in free Mg2+ concentration, from 10.25 to 0.6 mM when free Mg2+ is increased from 0.03 to 1.0 mM respectively. The Km for Mg2+ depends on the substrate concentration: the Km decreases from 0.52 to 0.14 mM from 0.25 to 0.75 mM-PP(i)-Mg2+ respectively. The extrapolated Km for Mg2+ in the absence of the substrate is 0.73 mM. Imidodiphosphate-Mg2+ and free Ca2+ were used as competitive inhibitors of substrate and activator respectively. The equilibrium binding kinetics suggest an ordered mechanism for the activator and the substrate: Mg2+ ions bind the enzyme before PP(i)-Mg2+ in the formation of the catalytic complex, membrane-bound pyrophosphatase-(Mg2+)-(PP(i)-Mg2+). PMID:1315519

  10. Listeriolysin O Membrane Damaging Activity Involves Arc Formation and Lineaction -- Implication for Listeria monocytogenes Escape from Phagocytic Vacuole

    PubMed Central

    Ruan, Yi; Rezelj, Saša; Bedina Zavec, Apolonija; Anderluh, Gregor; Scheuring, Simon

    2016-01-01

    Listeriolysin-O (LLO) plays a crucial role during infection by Listeria monocytogenes. It enables escape of bacteria from phagocytic vacuole, which is the basis for its spread to other cells and tissues. It is not clear how LLO acts at phagosomal membranes to allow bacterial escape. The mechanism of action of LLO remains poorly understood, probably due to unavailability of suitable experimental tools that could monitor LLO membrane disruptive activity in real time. Here, we used high-speed atomic force microscopy (HS-AFM) featuring high spatio-temporal resolution on model membranes and optical microscopy on giant unilamellar vesicles (GUVs) to investigate LLO activity. We analyze the assembly kinetics of toxin oligomers, the prepore-to-pore transition dynamics and the membrane disruption in real time. We reveal that LLO toxin efficiency and mode of action as a membrane-disrupting agent varies strongly depending on the membrane cholesterol concentration and the environmental pH. We discovered that LLO is able to form arc pores as well as damage lipid membranes as a lineactant, and this leads to large-scale membrane defects. These results altogether provide a mechanistic basis of how large-scale membrane disruption leads to release of Listeria from the phagocytic vacuole in the cellular context. PMID:27104344

  11. Listeriolysin O Membrane Damaging Activity Involves Arc Formation and Lineaction -- Implication for Listeria monocytogenes Escape from Phagocytic Vacuole.

    PubMed

    Ruan, Yi; Rezelj, Saša; Bedina Zavec, Apolonija; Anderluh, Gregor; Scheuring, Simon

    2016-04-01

    Listeriolysin-O (LLO) plays a crucial role during infection by Listeria monocytogenes. It enables escape of bacteria from phagocytic vacuole, which is the basis for its spread to other cells and tissues. It is not clear how LLO acts at phagosomal membranes to allow bacterial escape. The mechanism of action of LLO remains poorly understood, probably due to unavailability of suitable experimental tools that could monitor LLO membrane disruptive activity in real time. Here, we used high-speed atomic force microscopy (HS-AFM) featuring high spatio-temporal resolution on model membranes and optical microscopy on giant unilamellar vesicles (GUVs) to investigate LLO activity. We analyze the assembly kinetics of toxin oligomers, the prepore-to-pore transition dynamics and the membrane disruption in real time. We reveal that LLO toxin efficiency and mode of action as a membrane-disrupting agent varies strongly depending on the membrane cholesterol concentration and the environmental pH. We discovered that LLO is able to form arc pores as well as damage lipid membranes as a lineactant, and this leads to large-scale membrane defects. These results altogether provide a mechanistic basis of how large-scale membrane disruption leads to release of Listeria from the phagocytic vacuole in the cellular context. PMID:27104344

  12. Vesicular stomatitis virus glycoprotein mutations that affect membrane fusion activity and abolish virus infectivity.

    PubMed Central

    Fredericksen, B L; Whitt, M A

    1995-01-01

    We have introduced amino acid substitutions into two regions of the extracellular domain of the vesicular stomatitis virus (VSV) glycoprotein (G protein) and examined the effect of these mutations on protein transport, low-pH-induced stability of G protein oligomers, and membrane fusion activity. We suggested previously that the region between amino acids 118 and 139 may be important for the membrane fusion activity of G protein, on the basis of the characterization of a fusion-defective G protein mutant (M. A. Whitt, P. Zagouras, B. Crise, and J. K. Rose, J. Virol. 64:4907-4913, 1990). It has also been postulated by others that this region as well as the region between amino acids 181 and 212 may constitute putative internal fusion domains of VSV G protein. In this report, we show that three different amino acids substitutions between residues 118 and 139 (G-124-->E, P-127-->D, and A-133-->K) either altered or abolished low-pH-dependent membrane fusion activity. In contrast, substitutions between residues 192 and 212 resulted either in G proteins that had wild-type fusion activity or in mutant proteins in which the mutation prevented transport of G protein to the cell surface. Two of the substitutions between residues 118 and 139 (G-124-->E and P-127-->D) resulted in G proteins that were fusion defective at pH 5.7, although syncytia were observed after cells were treated with fusion buffer at pH 5.5, albeit at levels significantly less than that induced by wild-type G protein. Interestingly, when either G-124-->E or P-127-->D was incorporated into tsO45 virions, the resulting particles were not infectious, presumably because the viral envelope was not able to fuse with the proper intracellular membrane. These results support the hypothesis that the region between amino acids 118 and 139 is important for the membrane fusion activity of VSV G protein and may constitute an internal fusion domain. PMID:7853475

  13. Alkyl Galactofuranosides Strongly Interact with Leishmania donovani Membrane and Provide Antileishmanial Activity

    PubMed Central

    Suleman, Muhammad; Gangneux, Jean-Pierre; Legentil, Laurent; Belaz, Sorya; Cabezas, Yari; Manuel, Christelle; Dureau, Rémy; Sergent, Odile; Burel, Agnès; Daligault, Franck; Ferrières, Vincent

    2014-01-01

    We investigated the in vitro effects of four alkyl-galactofuranoside derivatives, i.e., octyl-β-d-galactofuranoside (compound 1), 6-amino-β-d-galactofuranoside (compound 2), 6-N-acetamido-β-d-galactofuranoside (compound 3), and 6-azido-β-d-galactofuranoside (compound 4), on Leishmania donovani. Their mechanism of action was explored using electron paramagnetic resonance spectroscopy (EPR) and nuclear magnetic resonance (NMR), and ultrastructural alterations were analyzed by transmission electron microscopy (TEM). Compound 1 showed the most promising effects by inhibiting promastigote growth at a 50% inhibitory concentration (IC50) of 8.96 ± 2.5 μM. All compounds exhibit low toxicity toward human macrophages. Compound 1 had a higher selectivity index than the molecule used for comparison, i.e., miltefosine (159.7 versus 37.9, respectively). EPR showed that compound 1 significantly reduced membrane fluidity compared to control promastigotes and to compound 3. The furanose ring was shown to support this effect, since the isomer galactopyranose had no effect on parasite membrane fluidity or growth. NMR showed a direct interaction of all compounds (greatest with compound 1, followed by compounds 2, 3, and 4, in descending order) with the promastigote membrane and with octyl-galactopyranose and octanol, providing evidence that the n-octyl chain was primarily involved in anchoring with the parasite membrane, followed by the putative crucial role of the furanose ring in the antileishmanial activity. A morphological analysis of compound 1-treated promastigotes by TEM revealed profound alterations in the parasite membrane and organelles, but this was not the case with compound 3. Quantification of annexin V binding by flow cytometry confirmed that compound 1 induced apoptosis in >90% of promastigotes. The effect of compound 1 was also assessed on intramacrophagic amastigotes and showed a reduction in amastigote growth associated with an increase of reactive oxygen

  14. Fatty acid composition of plasma lipids and erythrocyte membranes during simulated extravehicular activity

    NASA Astrophysics Data System (ADS)

    Skedina, M. A.; Katuntsev, V. P.; Buravkova, L. B.; Naidina, V. P.

    Ten subjects (from 27 to 41 years) have been participated in 32 experiments. They were decompressed from ground level to 40-35 kPa in altitude chamber when breathed 100% oxygen by mask and performed repeated cycles of exercises (3.0 Kcal/min). The intervals between decompressions were 3-5 days. Plasma lipid and erythrocyte membrane fatty acid composition was evaluated in the fasting venous blood before and immediately after hypobaric exposure. There were 7 cases decompression sickness (DCS). Venous gas bubbles (GB) were detected in 27 cases (84.4%). Any significant changes in the fatty acid composition of erythrocyte membranes and plasma didn't practically induce after the first decompression. However, by the beginning of the second decompression the total lipid level in erythrocyte membranes decreased from 54.6 mg% to 40.4 mg% in group with DCS symptoms and from 51.2 mg% to 35.2 mg% (p < 0.05) without DCS symptoms. In group with DCS symptoms a tendency to increased level of saturated fatty acids in erythrocyte membranes (16:0, 18:0), the level of the polyunsaturated linoleic fatty acid (18:2) and arachidonic acid (20:4) tended to be decreased by the beginning of the second decompression. Insignificant changes in blood plasma fatty acid composition was observed in both groups. The obtained biochemical data that indicated the simulated extravehicular activity (EVA) condition is accompanied by the certain changes in the blood lipid metabolism, structural and functional state of erythrocyte membranes, which are reversible. The most pronounced changes are found in subjects with DCS symptoms.

  15. Different metabolic activity in placental and reflected regions of the human amniotic membrane.

    PubMed

    Banerjee, Asmita; Weidinger, Adelheid; Hofer, Martin; Steinborn, Ralf; Lindenmair, Andrea; Hennerbichler-Lugscheider, Simone; Eibl, Johann; Redl, Heinz; Kozlov, Andrey V; Wolbank, Susanne

    2015-11-01

    Cells of the human amniotic membrane (hAM) have stem cell characteristics with low immunogenicity and anti-inflammatory properties. While hAM is an excellent source for tissue engineering, so far, its sub-regions have not been taken into account. We show that placental and reflected hAM differ distinctly in morphology and functional activity, as the placental region has significantly higher mitochondrial activity, however significantly less reactive oxygen species. Since mitochondria may participate in processes such as cell rescue, we speculate that amniotic sub-regions may have different potential for tissue regeneration, which may be crucial for clinical applications. PMID:26386652

  16. The membrane remodeling protein Pex11p activates the GTPase Dnm1p during peroxisomal fission

    PubMed Central

    Opalinski, Lukasz; Landgraf, Christiane; Costello, Joseph; Schrader, Michael; Krikken, Arjen M.; Knoops, Kèvin; Kram, Anita M.; Volkmer, Rudolf; van der Klei, Ida J.

    2015-01-01

    The initial phase of peroxisomal fission requires the peroxisomal membrane protein Peroxin 11 (Pex11p), which remodels the membrane, resulting in organelle elongation. Here, we identify an additional function for Pex11p, demonstrating that Pex11p also plays a crucial role in the final step of peroxisomal fission: dynamin-like protein (DLP)-mediated membrane scission. First, we demonstrate that yeast Pex11p is necessary for the function of the GTPase Dynamin-related 1 (Dnm1p) in vivo. In addition, our data indicate that Pex11p physically interacts with Dnm1p and that inhibiting this interaction compromises peroxisomal fission. Finally, we demonstrate that Pex11p functions as a GTPase activating protein (GAP) for Dnm1p in vitro. Similar observations were made for mammalian Pex11β and the corresponding DLP Drp1, indicating that DLP activation by Pex11p is conserved. Our work identifies a previously unknown requirement for a GAP in DLP function. PMID:25941407

  17. Human proximal tubule epithelial cells cultured on hollow fibers: living membranes that actively transport organic cations.

    PubMed

    Jansen, J; De Napoli, I E; Fedecostante, M; Schophuizen, C M S; Chevtchik, N V; Wilmer, M J; van Asbeck, A H; Croes, H J; Pertijs, J C; Wetzels, J F M; Hilbrands, L B; van den Heuvel, L P; Hoenderop, J G; Stamatialis, D; Masereeuw, R

    2015-01-01

    The bioartificial kidney (BAK) aims at improving dialysis by developing 'living membranes' for cells-aided removal of uremic metabolites. Here, unique human conditionally immortalized proximal tubule epithelial cell (ciPTEC) monolayers were cultured on biofunctionalized MicroPES (polyethersulfone) hollow fiber membranes (HFM) and functionally tested using microfluidics. Tight monolayer formation was demonstrated by abundant zonula occludens-1 (ZO-1) protein expression along the tight junctions of matured ciPTEC on HFM. A clear barrier function of the monolayer was confirmed by limited diffusion of FITC-inulin. The activity of the organic cation transporter 2 (OCT2) in ciPTEC was evaluated in real-time using a perfusion system by confocal microscopy using 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide (ASP(+)) as a fluorescent substrate. Initial ASP(+) uptake was inhibited by a cationic uremic metabolites mixture and by the histamine H2-receptor antagonist, cimetidine. In conclusion, a 'living membrane' of renal epithelial cells on MicroPES HFM with demonstrated active organic cation transport was successfully established as a first step in BAK engineering. PMID:26567716

  18. Interactions of membrane potential and cations in regulation of ciliary activity in Paramecium.

    PubMed

    Machemer, H

    1976-10-01

    Ciliary activity in Paramecium was investigated in different external solutions using techniques of voltage clamp and high frequency cinematography. An increase in the external concentration of K, Ca or Mg ions decreased the resting potential. It had no effect on ciliary activity. When the membrane potential was fixed, an increase in external Ca or Mg and, to a lesser extent, an increase in K concentration, raised the frequency of normal beating or decreased the frequency of reversed beating of the cilia. Similar effects resulted from membrane hyperpolarization with constant ionic conditions. Increase in concentration of Ca, but not of Mg or K, enhanced hyperpolarization-induced augmentation of ciliary frequency. Increase in Ca concentration also specifically augmented the delayed increase in inward current during rapid hyperpolarizing clamp. The results support the view that [Ca]i regulates the frequency and direction of ciliary beating. It is suggested that the insensitivity of the ciliary motor system to elevations of the external concentrations of ions results from compensation of their effects on [Ca]i. Depolarization itself appears to increase [Ca]i while elevation of the external ion concentrations at a fixed membrane potential appears to decrease [Ca]i. PMID:1003088

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

    PubMed

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

    2016-12-01

    The biological functions of cell membranes strongly influence the binding and transport of molecular species. We developed STC-1 cell line stably expressing the sweet taste receptor (T1R2/T1R3), and explored the possible correlation between sweeteners and membrane dipole potential of STC-1 cells. In this study, sweetener-induced dipole potential activation was elucidated using a fluorescence-based measurement technique, by monitoring the voltage sensitive probe Di-8-ANEPPS using a dual wavelength ratiometric approach. It indicated that the presence of sweeteners resulted in cell membrane dipole potential change, and interaction of artificial sweeteners with taste cells resulted in a greater reduction in potential compared with natural sweeteners. Our work presents a newly developed approach using a fluorescence-based measurement technique to study sweetener-induced dipole potential activation of STC-1 cells. This new approach could be used as a complementary tool to study the function of sweet taste receptors or other GPCRs and helps to understand the basis sweetness mechanism. PMID:27374594

  20. Antibacterial activity and in vitro evaluation of the biocompatibility of chitosan-based polysaccharide/polyester membranes.

    PubMed

    Wu, Chin-San; Hsu, Yi-Chiang; Liao, Hsin-Tzu; Cai, Yu-Xuan

    2015-12-10

    The antibacterial activity and biocompatibility of membranes of poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and chitosan (CS) (PHBV)/CS) were evaluated in this study. Maleic anhydride (MA)-grafted polyhydroxyalkanoate (PHBV-g-MA) was evaluated as an alternative to PHBV. Mouse tail skin fibroblasts (FBs) were seeded on two series of these films to assess cytocompatibility. Collagen and cell proliferation analyses indicated that PHBV, PHBV-g-MA and their composite membranes were biocompatible with respect to FB proliferation. However, FB proliferation, collagen production and the percentage of normal cells growing on PHBV/CS membranes were greater than those for PHBV-g-MA/CS membranes. Cell-cycle and apoptosis assays by FBs on the PHBV-series membrane samples were not affected by DNA content related to damage; i.e. rapid apoptosis/necrosis was not observed, demonstrating the potential of PHBV/CS or PHBV-g-MA/CS membranes for biomedical material applications. Moreover, CS-based polysaccharide enhanced the Escherichia coli (BCRC 10239) antibacterial activity of the membranes. Membranes of PHBV-g-MA or PHBV containing CS-based polysaccharide had better antibacterial activity. PMID:26428145

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

    NASA Astrophysics Data System (ADS)

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

    2015-03-01

    General adoption of advanced treatment protocols such as chronotherapy will hinge on progress in drug delivery technologies that provide precise temporal control of therapeutic release. Such innovation is also crucial to future medicine approaches such as telemedicine. Here we present a nanofluidic membrane technology capable of achieving active and tunable control of molecular transport through nanofluidic channels. Control was achieved through application of an electric field between two platinum electrodes positioned on either surface of a 5.7 nm nanochannel membrane designed for zero-order drug delivery. Two electrode configurations were tested: laser-cut foils and electron beam deposited thin-films, configurations capable of operating at low voltage (<=1.5 V), and power (100 nW). Temporal, reproducible tuning and interruption of dendritic fullerene 1 (DF-1) transport was demonstrated over multi-day release experiments. Conductance tests showed limiting currents in the low applied potential range, implying ionic concentration polarization (ICP) at the interface between the membrane's micro- and nanochannels, even in concentrated solutions (<=1 M NaCl). The ability of this nanotechnology platform to facilitate controlled delivery of molecules and particles has broad applicability to next-generation therapeutics for numerous pathologies, including autoimmune diseases, circadian dysfunction, pain, and stress, among others.General adoption of advanced treatment protocols such as chronotherapy will hinge on progress in drug delivery technologies that provide precise temporal control of therapeutic release. Such innovation is also crucial to future medicine approaches such as telemedicine. Here we present a nanofluidic membrane technology capable of achieving active and tunable control of molecular transport through nanofluidic channels. Control was achieved through application of an electric field between two platinum electrodes positioned on either surface of a 5

  2. The Immunosuppressive Activity of Amniotic Membrane Mesenchymal Stem Cells on T Lymphocytes

    PubMed Central

    Alikarami, Fatemeh; Yari, Fatemeh; Amirizadeh, Naser; Nikougoftar, Mahin; Jalili, Mohammad Ali

    2015-01-01

    Background: Mesenchymal Stem Cells (MSCs) are isolated from different sources like placenta. The placenta and its membranes like Amniotic Membrane (AM) are readily available and easy to work with. There is only limited knowledge on the immunomodulatory properties of human Amniotic Membrane-derived Mesenchymal Stem Cells (hAM-MSCs). The aim of this study was to survey the suppressive activity of hAM-MSCs on T lymphocytes in vitro. Methods: Human AMs were obtained after caesarean section births from healthy women. After enzymatic digestion, cells were cultured and hAM-MSCs were obtained. In addition, human T lymphocytes were isolated and co-cultured with hAM-MSCs for 72 hr in the presence or absence of phytohemagglutinin (PHA). Subsequently, proliferation of T cells was analyzed using BrdU and subsequently flow cytometry technique. Besides, the production of IL-4 and IFN-γ was examined by ELISA method. Additionally, the expression of activation markers (CD38, HLA-DR) was studied on T lymphocytes by flow cytometry technique. Results: It was revealed that hAM-MSCs could significantly suppress the proliferation of T lymphocytes (p≤0.01) and significantly decrease the production of IFN-γ by T cells (p<0.05). hAM-MSCs also down regulated the expression of activation markers on the surface of T lymphocytes, CD38 and HLA-DR. The difference was significant between the case and control samples (p<0.05). All the comparisons were carried out between the case (Tcell+PHA+hAM-MSCs) and control (Tcell+PHA) groups. Conclusion: In conclusion, hAM-MSCs could inhibit the (mitogen-activated) T cells even in the absence of blood monocytes. Besides, hAM-MSCs-mediated inhibition of T lymphocytes was combined with down regulation of activation markers. PMID:26306147

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

    SciTech Connect

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

    2008-12-15

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

  4. Antimicrobial properties of membrane-active dodecapeptides derived from MSI-78.

    PubMed

    Monteiro, Claudia; Fernandes, Mariana; Pinheiro, Marina; Maia, Sílvia; Seabra, Catarina L; Ferreira-da-Silva, Frederico; Costa, Fabíola; Reis, Salette; Gomes, Paula; Martins, M Cristina L

    2015-05-01

    Antimicrobial peptides (AMPs) are a class of broad-spectrum antibiotics known by their ability to disrupt bacterial membranes and their low tendency to induce bacterial resistance, arising as excellent candidates to fight bacterial infections. In this study we aimed at designing short 12-mer AMPs, derived from a highly effective and broad spectrum synthetic AMP, MSI-78 (22 residues), by truncating this peptide at the N- and/or C-termini while spanning its entire sequence with 1 amino acid (aa) shifts. These designed peptides were evaluated regarding antimicrobial activity against selected gram-positive Staphylococcus strains and the gram-negative Pseudomonas aeruginosa (P. aeruginosa). The short 12-mer peptide CEM1 (GIGKFLKKAKKF) was identified as an excellent candidate to fight P. aeruginosa infections as it displays antimicrobial activity against this strain and selectivity, with negligible toxicity to mammalian cells even at high concentrations. However, in general most of the short 12-mer peptides tested showed a reduction in antimicrobial activity, an effect that was more pronounced for gram-positive Staphylococcus strains. Interestingly, CEM1 and a highly similar peptide differing by only one aa-shift (CEM2: IGKFLKKAKKFG), showed a remarkably contrasting AMP activity. These two peptides were chosen for a more detailed study regarding their mechanism of action, using several biophysical assays and simple membrane models that mimic the mammalian and bacterial lipid composition. We confirmed the correlation between peptide helicity and antimicrobial activity and propose a mechanism of action based on the disruption of the bacterial membrane permeability barrier. PMID:25680229

  5. Study of FBAR response with variation in active area of membrane

    NASA Astrophysics Data System (ADS)

    Gill, Gurpreet Singh; Singh, Tarandip; Prasad, Mahanth

    2016-04-01

    In most of the communication devices such as filter, duplexer and oscillator, the need of acoustic resonator is the key part because of their small size and high performances. The design of a resonator based on three layers: (1) Bottom metal electrode such as Pt, Mo, Al and Au etc. (2) Piezoelectric layer such as ZnO, AlN and PZT etc. and (3) Top metal electrode. In this paper, the effects of active area on resonance frequency and impedance response of FBAR device have been studied. The FBAR devices having different membrane sizes, 150×150 µm2, 300×300 µm2, 450×450 µm2 and 600×600 µm2 were designed and simulated using COMSOL software Tool. The variation in resonance frequencies are found to be 2.62-2.65 GHz. Based on simulation results, one of the membrane having size, 300×300 µm2 has been fabricated for FBAR device.

  6. Broad spectrum antibacterial and antifungal polymeric paint materials: synthesis, structure-activity relationship, and membrane-active mode of action.

    PubMed

    Hoque, Jiaul; Akkapeddi, Padma; Yadav, Vikas; Manjunath, Goutham B; Uppu, Divakara S S M; Konai, Mohini M; Yarlagadda, Venkateswarlu; Sanyal, Kaustuv; Haldar, Jayanta

    2015-01-28

    Microbial attachment and subsequent colonization onto surfaces lead to the spread of deadly community-acquired and hospital-acquired (nosocomial) infections. Noncovalent immobilization of water insoluble and organo-soluble cationic polymers onto a surface is a facile approach to prevent microbial contamination. In the present study, we described the synthesis of water insoluble and organo-soluble polymeric materials and demonstrated their structure-activity relationship against various human pathogenic bacteria including drug-resistant strains such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant enterococci (VRE), and beta lactam-resistant Klebsiella pneumoniae as well as pathogenic fungi such as Candida spp. and Cryptococcus spp. The polymer coated surfaces completely inactivated both bacteria and fungi upon contact (5 log reduction with respect to control). Linear polymers were more active and found to have a higher killing rate than the branched polymers. The polymer coated surfaces also exhibited significant activity in various complex mammalian fluids such as serum, plasma, and blood and showed negligible hemolysis at an amount much higher than minimum inhibitory amounts (MIAs). These polymers were found to have excellent compatibility with other medically relevant polymers (polylactic acid, PLA) and commercial paint. The cationic hydrophobic polymer coatings disrupted the lipid membrane of both bacteria and fungi and thus showed a membrane-active mode of action. Further, bacteria did not develop resistance against these membrane-active polymers in sharp contrast to conventional antibiotics and lipopeptides, thus the polymers hold great promise to be used as coating materials for developing permanent antimicrobial paint. PMID:25541751

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

    SciTech Connect

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

    2010-03-26

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

  8. Membrane IL1α Inhibits the Development of Hepatocellular Carcinoma via Promoting T- and NK-cell Activation.

    PubMed

    Lin, Dandan; Lei, Lei; Liu, Yonghao; Zhang, Yinsheng; Hu, Bo; Bao, Guangming; Song, Yuan; Jin, Ziqi; Liu, Chunliang; Mei, Yu; Sandikin, Dedy; Wu, Yan; Zhao, Lixiang; Yu, Xiao; Liu, Haiyan

    2016-06-01

    Hepatocellular carcinoma is a worldwide health problem with limited treatment options and poor prognosis. Inflammation associated with liver injury and hepatocyte regeneration can lead to fibrosis, cirrhosis, and eventually, hepatocellular carcinoma. IL1α is one of the most important inflammatory cytokines involved in inflammation and tumor development. IL1α presents as multiple forms in vivo, including precursor, propiece, membrane, and secreted forms, and their functions have been thought to be different. The role of membrane IL1α in hepatocellular carcinoma tumorigenesis is still not clear. Here, we examined the functions of membrane IL1α in murine hepatocellular carcinoma models. We found that membrane IL1α potently inhibited hepatocellular carcinoma tumor growth. Further studies showed that membrane IL1α promoted T- and natural killer (NK)-cell activation in vivo IFNγ production by CD8(+) T and NK cells was also increased as a result of membrane IL1α expression. Moreover, the cytotoxicity of the CTL and NK cells was also enhanced by membrane IL1α expression. Furthermore, in vitro studies demonstrated that membrane IL1α could directly activate T cells and NK cells in a cell contact-dependent manner. Conversely, depletion of both CD8(+) T and NK cells suppressed the antitumor activity of membrane IL1α. Our studies demonstrated that membrane IL1α could promote antitumor immune responses through activation of T and NK cells. Thus, our findings provide new insights of IL1α functions during hepatocellular carcinoma development. Cancer Res; 76(11); 3179-88. ©2016 AACR. PMID:27206848

  9. A Placental Polypeptide Activator of a Membranous Protein Kinase and Its Relation to Histone 1

    NASA Astrophysics Data System (ADS)

    Abdel-Ghany, M.; Riegler, C.; Racker, E.

    1984-12-01

    Crude transforming growth factor preparations of placenta contain a polypeptide that is required for the activity of a protein kinase that has been purified from plasma membrane preparations of Ehrlich ascites tumor cells. The kinase activator has been separated from transforming growth factor β by reversed-phase HPLC and affinity chromatography. Like the transforming growth factor, it is heat stable and trypsin labile, but it is not inactivated by dithiothreitol. In sodium dodecyl sulfate/polyacrylamide gel electrophoresis the purified preparation shows a major double band at about 31,000 daltons. Comparisons of electrophoretic mobility, protein kinase stimulatory activity, and cross-reactivity with an antibody against histone 1 suggest that the placental activator is identical with histone 1.

  10. Temporin L: antimicrobial, haemolytic and cytotoxic activities, and effects on membrane permeabilization in lipid vesicles.

    PubMed Central

    Rinaldi, Andrea C; Mangoni, Maria Luisa; Rufo, Anna; Luzi, Carla; Barra, Donatella; Zhao, Hongxia; Kinnunen, Paavo K J; Bozzi, Argante; Di Giulio, Antonio; Simmaco, Maurizio

    2002-01-01

    The temporins are a family of small, linear antibiotic peptides with intriguing biological properties. We investigated the antibacterial, haemolytic and cytotoxic activities of temporin L (FVQWFSKFLGRIL-NH2), isolated from the skin of the European red frog Rana temporaria. The peptide displayed the highest activity of temporins studied to date, against both human erythrocytes and bacterial and fungal strains. At variance with other known temporins, which are mainly active against Gram-positive bacteria, temporin L was also active against Gram-negative strains such as Pseudomonas aeruginosa A.T.C.C. 15692 and Escherichia coli D21 at concentrations comparable with those that are microbiocidal to Gram-positive bacteria. In addition, temporin L was cytotoxic to three different human tumour cell lines (Hut-78, K-562 and U-937), causing a necrosis-like cell death, although sensitivity to the peptide varied markedly with the specific cell line tested. A study of the interaction of temporin L with liposomes of different lipid compositions revealed that the peptide causes perturbation of bilayer integrity of both neutral and negatively charged membranes, as revealed by the release of a vesicle-encapsulated fluorescent marker, and that the action of the peptide is modulated to some extent by membrane lipid composition. In particular, the presence of negatively charged lipids in the model bilayer inhibits the lytic power of temporin L. We also show that the release of fluorescent markers caused by temporin L is size-dependent and that the peptide does not have a detergent-like effect on the membrane, suggesting that perturbation of bilayer organization takes place on a local scale, i.e. through the formation of pore-like openings. PMID:12133008

  11. Membrane Binding Events in the Initiation and Propagation Phases of Tissue Factor-initiated Zymogen Activation under Flow*

    PubMed Central

    Haynes, Laura M.; Dubief, Yves C.; Mann, Kenneth G.

    2012-01-01

    This study investigates the dynamics of zymogen activation when both extrinsic tenase and prothrombinase are assembled on an appropriate membrane. Although the activation of prothrombin by surface-localized prothrombinase is clearly mediated by flow-induced dilutional effects, we find that when factor X is activated in isolation by surface-localized extrinsic tenase, it exhibits characteristics of diffusion-mediated activation in which diffusion of substrate to the catalytically active region is rate-limiting. When prothrombin and factor X are activated coincident with each other, competition for available membrane binding sites masks the diffusion-limiting effects of factor X activation. To verify the role of membrane binding in the activation of factor X by extrinsic tenase under flow conditions, we demonstrate that bovine lactadherin competes for both factor X and Xa binding sites, limiting factor X activation and forcing the release of bound factor Xa from the membrane at a venous shear rate (100 s−1). Finally, we present steady-state models of prothrombin and factor X activation under flow showing that zymogen and enzyme membrane binding events further regulate the coagulation process in an open system representative of the vasculature geometry. PMID:22187432

  12. Generation, modulation and maintenance of the plasma membrane asymmetric phospholipid composition in yeast cells during growth: their relation to surface potential and membrane protein activity.

    PubMed

    Cerbón, J; Calderón, V

    1995-04-12

    During growth a cyclic exposure of anionic phospholipids to the external surface of the plasma membrane was found. The surface charge density (sigma) increased gradually reaching a maximum in the first 5 h of growth and returned gradually to their initial value at the end of the logarithmic phase of growth (10-12 h). Phosphatidylinositol, that determines to a large extent the magnitude of the sigma, increased 83% in the yeast cells during the first 4 h of growth and returned gradually to their initial level at 10-12 h. During the stationary phase (12-24 h), both sigma and the anionic/zwitterionic phospholipid ratio, remained without any significant variation. The high-affinity H-linked glutamate transport system that behaves as a sensor of the changes in the membrane surface potential (phi) increased its activity in the first 5 h and then decreased it, following with great accuracy the sigma variations and remained without changes during the stationary phase of growth. The phosphatidylserine (PS) relative concentration in the cells (9.0%) did not significantly change during the whole growth curve, but their asymmetric distribution varied, contributing to the changes in sigma. PS facing the outer membrane surface increased 2.45-times during the first 5 h of growth and then returned to their original value at the end of the log phase (12 h). Phosphatidylcholine (PC) remained constant during the whole growth curve (50%), while phosphatidylethanolamine (PE) decreased 3-fold in the first 4 h and then increased to its original value at 10 h. Interestingly, PE at the outer membrane surface remained constant (3% of the total phospholipids) during the whole growth curve. During growth yeast cells change their phospholipid composition originating altered patterns of the plasma membrane phospholipid composition and IN-OUT distribution. This dynamic asymmetry is involved in the regulation of the surface potential and membrane protein activity. PMID:7718598

  13. Study on the effects of nylon-chitosan-blended membranes on the spheroid-forming activity of human melanocytes.

    PubMed

    Lin, Sung-Jan; Hsiao, Wen-Chu; Jee, Shiou-Hwa; Yu, Hsin-Su; Tsai, Tsen-Fang; Lai, Juin-Yih; Young, Tai-Horng

    2006-10-01

    Though reported limitedly in tissue engineering, modification of cellular functions can be achieved by culturing them into multicellular spheroids. We have shown melanocytes form spheroids on chitosan surface. However, how biomaterials promote spheroid formation has never been systemically investigated. In this work, nylon, which inhibits melanocyte spheroid formation, and chitosan, which promotes melanocyte spheroid formation, are used to prepare nylon/chitosan-blended membranes. Membranes composed of pure nylon, pure chitosan and various ratios of nylon and chitosan are employed to examine their effects on spheroid formation. Melanocytes show better adhesion to nylon membranes than that to chitosan membranes. In blended membranes, as more nylon is incorporated, cell adhesion increases and the trend for spheroid formation decreases. Melanocytes can only form spheroids on membranes with poorer cell adhesion. Examining the surface of the blended membranes shows phase separation of nylon and chitosan. As nylon content increases, the nylon phase on the membrane surface increases and thereby enhances cell adhesion. The opposite trend for cell adhesion and spheroid formation substantiates our hypothesis of spheroid formation on biomaterials: a balance between cell-substrate interaction and cell-cell interaction. The decrease in cell-substrate interaction tilts the balance to a state more favorable for spheroid formation. Our work can serve as a model to investigate the relative strengths of cell-cell and cell-substrate interactions and also pave way to design blended membranes with desired physical properties while preserving the spheroid-forming activity. PMID:16777216

  14. Net Increase of platelet membrane tyrosine specific-protein kinase activity by phorbol myristate acetate

    SciTech Connect

    Ishihara, Noriko; Sakamoto, Hikaru; Iwama, Minako; Kobayashi, Bonro )

    1990-01-01

    Tyrosine protein kinase (TPK) activity in rabbit platelets after stimulation by phorbol myristate acetate (PMA) or thrombin was directly estimated by {sup 32}P incorporation from ({gamma}-{sup 32})ATP into synthetic peptide angiotensin II. By PMA-treatment a net increase of TPK activity was obtained, while thrombin acted on the TPK quickly but stimulation was limited within the range attained by the control after lengthy incubation. The responsive TPK to these stimulators was localized mainly in membrane but much less in cytosol. The specific activity of the particulate TPK was low in the sonicate of control ice cold platelets but increased about 6-fold when the platelets were incubated at 37{degree}C. On a brief contact of platelets with PMA at 37{degrees}C the TPK was fully activated and reached a maximum value about 130% of the control. Determination of phosphotyrosine phosphatase in the stimulated platelet sonicate revealed that its participation in the above described increase of {sup 32}P-incorporation was meagre. The quick response suggested a possible role of TPK in the signal transduction through the platelet cell membrane.

  15. Acetylcholinesterase activity of synaptic plasma membranes during ageing: effect of L-acetylcarnitine.

    PubMed

    Gorini, A; Ghigini, B; Villa, R F

    1996-01-01

    A physiopathological role for acetylcholine (ACh) was hypothesized during ageing and related neurodegenerative diseases, e.g. dementia. This research was aimed to study acetylcholinesterase (AChE) activity during development and ageing of the frontal cerebral cortex of 4-, 8-, 12-, 16-, 20- and 24-month-old rats. This study was performed on synaptic plasma membranes, the specific subcellular compartment where the enzyme is located in vivo both in control animals and after in vivo acute treatment with L-acetylcarnitine. Maximum AChE activity was unaffected by age, and L-acetylcarnitine treatment increased enzyme activity in synaptic plasma membranes of 8-month-old rats. A comprehensive analysis of these results suggests: (a) the observed alterations in protein can substantially affect neurochemical data if results are presented as specific activities per unit protein; (b) energy metabolism plays the major role in the disturbed ACh metabolism during ageing and (c) the understanding of the mode of action of L-acetylcarnitine in treatment of dementia. PMID:8740629

  16. CARBONIC ANHYDRASE ACTIVITY OF INTEGRAL-FUNCTIONAL COMPLEXES OF THYLAKOID MEMBRANES OF SPINACH CHLOROPLASTS.

    PubMed

    Semenihin, A V; Zolotareva, O K

    2015-01-01

    Isolated thylakoid membranes were disrupted by treatment with nonionic detergents digitonin or dodecyl maltoside. Solubilized polypeptide complexes were separated by native gel charge shift electrophoresis. The position of ATP-synthase complex and its isolated catalytic part (CF1) within gel was determined using the color reaction for ATPase activity. Due to the presence of cytochromes, the red band in unstained gels corresponded to the cytochrome b6f complex. Localization of the cytochrome b6f complex, ATP synthase and coupling CF1 in the native gel was confirmed by their subunit composition determined after SDS-electrophoretic analysis. Carbonic anhydrase (CA) activity in polypeptide zones of PS II, cytochrome b6f complex, and ATP-synthase CF1 was identified in native gels using indicator bromothymol blue. CA activity of isolated CF1 in solution was determined by infrared gas analysis as the rate of bicarbonate dehydration. The water-soluble acetazolamide, an inhibitor of CA, unlike lipophilic ethoxyzolamide inhibited CA activity of CF1 Thus, it was shown for the first time that ATP-synthase has a component which is capable of catalyzing the interconversion of forms of carbonic acid associated with proton exchange. The data obtained suggest the presence of multiple forms of carbonic anhydrase in the thylakoid membranes of spinach chloroplasts and confirm their involvement in the proton transfer to the ATP synthase. PMID:26502699

  17. Activation of Plant Plasma Membrane Ca2+-Permeable Channels by Race-Specific Fungal Elicitors.

    PubMed Central

    Gelli, A.; Higgins, V. J.; Blumwald, E.

    1997-01-01

    The response of plant cells to invading pathogens is regulated by fluctuations in cytosolic Ca2+ levels that are mediated by Ca2+-permeable channels located at the plasma membrane of the host cell. The mechanisms by which fungal elicitors can induce Ca2+ uptake by the host cell were examined by the application of conventional patch-clamp techniques. Whole-cell and single-channel experiments on tomato (Lycopersicon esculentum L.) protoplasts revealed a race-specific fungal elicitor-induced activation of a plasma membrane Ca2+-permeable channel. The presence of the fungal elicitor resulted in a greater probability of channel opening. Guanosine 5[prime]-[[beta]-thio]diphosphate, a GDP analog that locks heterotrimeric G-proteins into their inactivated state, abolished the channel activation induced by the fungal elicitor, whereas guanosine 5[prime][[gamma]-thio]triphosphate, a nonhydrolyzable GTP analog that locks heterotrimeric G-proteins into their activated state, produced an effect similar to that observed with the fungal elicitor. Mastoparan, which stimulates GTPase activity, mimicked the effect of GTP[[gamma

  18. Isoprenoid addition to Ras protein is the critical modification for its membrane association and transforming activity.

    PubMed Central

    Kato, K; Cox, A D; Hisaka, M M; Graham, S M; Buss, J E; Der, C J

    1992-01-01

    We have introduced a variety of amino acid substitutions into carboxyl-terminal CA1A2X sequence (C = cysteine; A = aliphatic; X = any amino acid) of the oncogenic [Val12]Ki-Ras4B protein to identify the amino acids that permit Ras processing (isoprenylation, proteolysis, and carboxyl methylation), membrane association, and transformation in cultured mammalian cells. While all substitutions were tolerated at the A1 position, substitutions at A2 and X reduced transforming activity. The A2 residue was important for both isoprenylation and AAX proteolysis, whereas the X residue dictated the extent and specificity of isoprenoid modification only. Differences were observed between Ras processing in living cells and farnesylation efficiency in a cell-free system. Finally, one farnesylated mutant did not undergo either proteolysis or carboxyl methylation but still displayed efficient membrane association (approximately 50%) and transforming activity, indicating that farnesylation alone can support Ras transforming activity. Since both farnesylation and carboxyl methylation are critical for yeast a-factor biological activity, the three CAAX-signaled modifications may have different contributions to the function of different CAAX-containing proteins. Images PMID:1631135

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

    PubMed

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

    2015-07-23

    Treating bacterial biofilms with conventional antibiotics is limited due to ineffectiveness of the drugs and higher propensity to develop bacterial resistance. Development of new classes of antibacterial therapeutics with alternative mechanisms of action has become imperative. Herein, we report the design, synthesis, and biological evaluations of novel membrane-active small molecules featuring two positive charges, four nonpeptidic amide groups, and variable hydrophobic/hydrophilic (amphiphilic) character. The biocides synthesized via a facile methodology not only displayed good antibacterial activity against wild-type bacteria but also showed high activity against various drug-resistant bacteria such as methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecium (VRE), and β-lactam-resistant Klebsiella pneumoniae. Further, these biocides not only inhibited the formation of biofilms but also disrupted the established S. aureus and E. coli biofilms. The membrane-active biocides hindered the propensity to develop bacterial resistance. Moreover, the biocides showed negligible toxicity against mammalian cells and thus bear potential to be used as therapeutic agents. PMID:26102297

  20. Para-aminobenzamidine linked regenerated cellulose membranes for plasminogen activator purification: Effect of spacer arm length and ligand density

    PubMed Central

    Fasoli, Ezio; Reyes, Yiaslin Ruiz; Guzman, Osiris Martinez; Rosado, Alexandra; Cruz, Vivian Rodriguez; Borges, Amaris; Martinez, Edmarie; Bansal, Vibha

    2013-01-01

    Despite membrane-based separations offering superior alternative to packed bed chromatographic processes, there has been a substantial lacuna in their actual application to separation processes. One of the major reasons behind this is the lack of availability of appropriately modified or end-group modifiable membranes. In this paper, an affinity membrane was developed using a commercially available serine protease inhibitor, para-aminobenzamidine (pABA). The membrane modification was optimized for protein binding capacity by varying: i) the length of the spacer arm (SA; 5-atoms, 7-atoms, and 14-atoms) linking the ligand to membrane surface; ii) the affinity ligand (pABA) density on membrane surface (5–25 nmoles per cm2). Resulting membranes were tested for their ability to bind plasminogen activators (PAs) from mono- and multi- component systems in batch mode. The membrane containing pABA linked through 7-atoms SA but similar ligand density as in the case of 5- or 14- atoms long SA was found to bind up to 1.6-times higher amounts of PA per nmole of immobilized ligand from conditioned HeLa cell culture media. However, membranes with similar ligand densities but different lengths of SA, showed comparable binding capacities in monocomponent system. In addition, the length of SA did not affect the selectivity of the ligand for PA. A clear inverse linear correlation was observed between ligand density and binding capacity until the point of PA binding optima was reached (11±1.0 nmoles per cm2) in mono- and multi- component systems for 7- as well as 14- atoms SA. Up to 200-fold purification was achieved in a single step separation of PA from HeLa conditioned media using these affinity membranes. The issues of ligand leaching and reuse of the membranes were also investigated. An extensive regeneration procedure allowed the preservation of approximately 95% of the PA binding capacity of the membranes even after five cycles of use. PMID:23703544

  1. Oriented Circular Dichroism: A Method to Characterize Membrane-Active Peptides in Oriented Lipid Bilayers.

    PubMed

    Bürck, Jochen; Wadhwani, Parvesh; Fanghänel, Susanne; Ulrich, Anne S

    2016-02-16

    The structures of membrane-bound polypeptides are intimately related to their functions and may change dramatically with the lipid environment. Circular dichroism (CD) is a rapid analytical method that requires relatively low amounts of material and no labeling. Conventional CD is routinely used to monitor the secondary structure of peptides and proteins in solution, for example, in the presence of ligands and other binding partners. In the case of membrane-active peptides and transmembrane proteins, these measurements can be applied to, and remain limited to, samples containing detergent micelles or small sonicated lipid vesicles. Such traditional CD analysis reveals only secondary structures. With the help of an oriented circular dichroism (OCD) setup, however, based on the preparation of macroscopically oriented lipid bilayers, it is possible to address the membrane alignment of a peptide in addition to its conformation. This approach has been mostly used for α-helical peptides so far, but other structural elements are conceivable as well. OCD analysis relies on Moffitt's theory, which predicts that the electronic transition dipole moments of the backbone amide bonds in helical polypeptides are polarized either parallel or perpendicular to the helix axis. The interaction of the electric field vector of the circularly polarized light with these transitions results in an OCD spectrum of a membrane-bound α-helical peptide, which exhibits a characteristic line shape and reflects the angle between the helix axis and the bilayer normal. For parallel alignment of a peptide helix with respect to the membrane surface (S-state), the corresponding "fingerprint" CD band around 208 nm will exhibit maximum negative amplitude. If the helix changes its alignment via an obliquely tilted (T-state) to a fully inserted transmembrane orientation (I-state), the ellipticity at 208 nm decreases and the value approaches zero due to the decreased interactions between the field and the

  2. Retaining activity of enzymes after capture and extraction within a single-drop of biological fluid using immunoaffinity membranes.

    PubMed

    Shimazaki, Youji; Sato, Yuki

    2016-05-15

    The purpose of this study was the measurement of enzyme activity within a single-drop of biological fluid after micropurification. Esterase and lactate dehydrogenase (LDH) retained their enzymatic activities after being captured by membrane-immobilized antibodies, which were prepared by non-denaturing two-dimensional electrophoresis, transferred to polyvinylidene difluoride and then stained by Ponceau S. The activities of both enzymes were also measured after being captured by antibodies and biotinylated antibodies bound to membrane-immobilized protein A or avidin, respectively. After esterase and LDH were captured from biological samples by membrane-immobilized protein A or avidin, their activities were semi-quantitatively measured on the surface of the membrane using fluorescence determination. More than 51% of enzyme activities were retained even after the enzymes were captured by biotinylated antibody bound to membrane-immobilized avidin and eluted by rinsing with 5μL of 1% Triton X-100, compared with the activities of the enzyme on the immunoaffinity membrane. PMID:26776499

  3. Polyurethane Ionophore-Based Thin Layer Membranes for Voltammetric Ion Activity Sensing.

    PubMed

    Cuartero, Maria; Crespo, Gaston A; Bakker, Eric

    2016-06-01

    We report on a plasticized polyurethane ionophore-based thin film material (of hundreds of nanometer thickness) for simultaneous voltammetric multianalyte ion activity detection triggered by the oxidation/reduction of an underlying poly(3-octylthiophene) film. This material provides excellent mechanical, physical, and chemical robustness compared to other polymers. Polyurethane films did not exhibit leaching of lipophilic additives after rinsing with a direct water jet and exhibited resistance to detachment from the underlying electrode surface, resulting in a voltammetric current response with less than <1.5% RSD variation (n = 50). In contrast, plasticized poly(vinyl chloride), polystyrene, and poly(acrylate) ionophore-based membranes of the same thickness and composition exhibited a significant deterioration of the signal after identical treatment. While previously reported works emphasized fundamental advancement of multi-ion detection with multi-ionophore-based thin films, polyurethane thin membranes allow one to achieve real world measurements without sacrificing analytical performance. Indeed, polyurethane membranes are demonstrated to be useful for the simultaneous determination of potassium and lithium in undiluted human serum and blood with attractive precision. PMID:27187779

  4. Regulating the Membrane Transport Activity and Death of Cells via Electroosmotic Manipulation.

    PubMed

    Hui, Tsz Hin; Kwan, Kin Wah; Chun Yip, Timothy Tak; Fong, Hong Wai; Ngan, Kai Cheong; Yu, Miao; Yao, Shuhuai; Wan Ngan, Alfonso Hin; Lin, Yuan

    2016-06-21

    Although the volume of living cells has been known to heavily influence their behavior and fate, a method allowing us to control the cell size in a programmable manner is still lacking. Here, we develop a technique in which precise changes in the cellular volume can be conveniently introduced by varying the voltage applied across a Nafion membrane that separates the culture medium from a reservoir. It is found that, unlike sudden osmotic shocks, active ion transport across the membrane of leukemia K562 cells will not be triggered by a gradual change in the extracellular osmolarity. Furthermore, when subjected to the same applied voltage, different lung and nasopharyngeal epithelial cancer cells will undergo larger volumetric changes and have a 5-10% higher death rate compared to their normal counterparts. We show that such distinct response is largely caused by the overexpression of aquaporin-4 in tumor cells, with knockout of this water channel protein resulting in a markedly reduced change in the cellular volume. Finally, by taking into account the exchange of water/ion molecules across the Nafion film and the cell membrane, a theoretical model is also proposed to describe the voltage-induced size changes of cells, which explain our experimental observations very well. PMID:27332135

  5. Cytolytic activity in T cell clones derived from human synovial rheumatoid membrane: inhibition by synovial fluid.

    PubMed Central

    Miltenburg, A M; Van Laar, J M; De Kuiper, P; Daha, M R; Breedveld, F C

    1990-01-01

    A panel of T cell clones was derived from the synovial membrane of a patient with rheumatoid arthritis (RA). We investigated whether T cell clones with cytolytic properties were present and whether T cell cytotoxicity was influenced by the presence of synovial fluid. These issues were studied using anti-CD3 and lectin-induced cytotoxicity assays. The majority of the T cell clones derived from the synovial membrane showed cytotoxic properties although non-cytotoxic clones were also found. Three clones (N11, N6 and N15) showed strong cytotoxicity (more than 40% lysis at an effector-to-target cell ratio of 10:1) whereas three clones (N16, N4 and N14) were non-cytotoxic (less than 20% lysis at an effector-to-target cell ratio of 10:1). The induction of cytotoxicity in the anti-CD3-driven system was shown to be dependent on the dose of anti-CD3 present. When synovial fluid was added to these assays a strong inhibition of cytotoxicity was found. This inhibition of cytotoxicity was found with synovial fluid samples of RA patients, as well as with non-RA synovial fluids. Both anti-CD3 and lectin-dependent cytotoxicity assays were strongly inhibited. In conclusion, T cell clones with cytotoxic activity can be isolated from rheumatoid synovial membrane. In the presence of synovial fluid these cytotoxic cells are inhibited to exert their cytotoxic function. PMID:2148285

  6. Barrier properties of poly(vinyl alcohol) membranes containing carbon nanotubes or activated carbon.

    PubMed

    Surdo, Erin M; Khan, Iftheker A; Choudhury, Atif A; Saleh, Navid B; Arnold, William A

    2011-04-15

    Carbon nanotube addition has been shown to improve the mechanical properties of some polymers. Because of their unique adsorptive properties, carbon nanotubes may also improve the barrier performance of polymers used in contaminant containment. This study compares the barrier performance of poly(vinyl alcohol) (PVA) membranes containing single-walled carbon nanotubes (SWCNTs) to that for PVA containing powdered activated carbon (PAC). Raw and surface-functionalized versions of each sorbent were tested for their abilities to adsorb 1,2,4-trichlorobenzene and Cu(2+), representing the important hydrophobic organic and heavy metal contaminant classes, as they diffused across the PVA. In both cases, PAC (for 1,2,4-trichlorobenzene) and functionalized PAC (for Cu(2+)) outperformed SWCNTs on a per mass basis by trapping more of the contaminants within the barrier membrane. Kinetics of sorption are important in evaluating barrier properties, and poor performance of SWCNT-containing membranes as 1,2,4-TCB barriers is attributed to kinetic limitations. PMID:21349636

  7. Effect of powdered activated carbon on integrated submerged membrane bioreactor-nanofiltration process for wastewater reclamation.

    PubMed

    Woo, Yun Chul; Lee, Jeong Jun; Shim, Wang-Geun; Shon, Ho Kyong; Tijing, Leonard D; Yao, Minwei; Kim, Han-Seung

    2016-06-01

    The aim of this study was to determine the effect of powdered activated carbon (PAC) on the overall performance of a submerged membrane bioreactor (SMBR) system integrated with nanofiltration (NF) for wastewater reclamation. It was found that the trans-membrane pressure of SMBR increased continuously while that of the SMBR with PAC was more stable, mainly because water could still pass through the PACs and membrane even though foulants adhered on the PAC surface. The presence of PAC was able to mitigate fouling in SMBR as well as in NF. SMBR-NF with PAC obtained a higher flux of 8.1 LMH compared to that without PAC (6.6 LMH). In addition, better permeate quality was obtained with S