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At least 55 records · Page 3

Fluorescence Studies of Protein Crystallization Interactions

We are investigating protein-protein interactions in under- and over-saturated crystallization solution conditions using fluorescence methods. The use of fluorescence requires fluorescent derivatives where the probe does not markedly affect the crystal packing. A number of chicken egg white lysozyme (CEWL) derivatives have been prepared, with the probes covalently attached to one of two different sites on the protein molecule; the side chain carboxyl of ASP 101, within the active site cleft, and the N-terminal amine. The ASP 101 derivatives crystallize while the N-terminal amine derivatives do not. However, the N-terminal amine is part of the contact region between adjacent 43 helix chains, and blocking this site does would not interfere with formation of these structures in solution. Preliminary FRET data have been obtained at pH 4.6, 0.1M NaAc buffer, at 5 and 7% NaCl, 4 C, using the N-terminal bound pyrene acetic acid (PAA, Ex 340 nm, Em 376 nm) and ASP 101 bound Lucifer Yellow (LY, Ex 425 nm, Em 525 nm) probe combination. The corresponding Csat values are 0.471 and 0.362 mg/ml (approximately 3.3 and approximately 2.5 x 10 (exp 5) M respectively), and all experiments were carried out at approximately Csat or lower total protein concentration. The data at both salt concentrations show a consistent trend of decreasing fluorescence yield of the donor species (PAA) with increasing total protein concentration. This decrease is apparently more pronounced at 7% NaCl, consistent with the expected increased intermolecular interactions at higher salt concentrations (reflected in the lower solubility). The estimated average distance between protein molecules at 5 x 10 (exp 6) M is approximately 70 nm, well beyond the range where any FRET can be expected. The calculated RO, where 50% of the donor energy is transferred to the acceptor, for the PAA-CEWL * LY-CEWL system is 3.28 nm, based upon a PAA-CEWL quantum efficiency of 0.41.

Pusey, Marc L.↗

Preparation and Preliminary Characterization of Crystallizing Fluorescent Derivatives of Chicken Egg White Lysozyme

Fluorescence is one of the most versatile and powerful tools for the study of macromolecules. While most proteins are intrinsically fluorescent, working at crystallization concentrations require the use of covalently prepared derivatives added as tracers. This approach requires derivatives that do not markedly affect the crystal packing. We have prepared fluorescent derivatives of chicken egg white lysozyme with probes bound to one of two different sites on the protein molecule. Lucifer yellow and 5-(2-aminoethyl)aminonapthalene-1-sulfonic acid (EDANS) have been attached to the side chain carboxyl of Asp(sup 101) using a carbodiimide coupling procedure. Asp(sup 101) lies within the active site cleft, and it is believed that the probes are "buried" within that cleft. Lucifer yellow and MANS probes with iodoacetamide reactive groups have been bound to His(sup 15), located on the "back side" of the molecule relative to the active site. All the derivatives fluoresce in the solution and the crystalline states. Fluorescence characterization has focused on determination of binding effects on the probe quantum yield, lifetime, absorption and emission spectra, and quenching by added solutes. Quenching studies show that, as postulated, the Asp(sup 101)-bound probes are partially sheltered from the bulk solution by their location within the active site cleft. Probes bound to His(sup 15) have quenching constants about equal to those for the free probes, indicating that this site is highly exposed to the bulk solution.

Sumida, John↗

Preparation and Preliminary Characterization of Crystallizing Fluorescent Derivatives of Chicken Egg White Lysozyme

Fluorescence is one of the most versatile and powerful tools for the study of macromolecules. While most proteins are intrinsically fluorescent, working at crystallization concentrations require the use of covalently prepared derivatives added as tracers. This approach requires derivatives that do not markedly affect the crystal packing. We have prepared fluorescent derivatives of chicken egg white lysozyme with probes bound to one of two different sites on the protein molecule. Lucifer yellow and 5-(2-aminoethyl)aminonapthalene-i-sulfonic acid (EDANS) have been attached to the side chain carboxyl of Asp(sup 101) using a carbodiimide coupling procedure. Asp(sup 101) lies within the active site cleft, and it is believed that the probes are 'buried' within that cleft. Lucifer yellow and MANS probes with iodoacetamide reactive five groups have been bound to His(sup 15), located on the 'back side' of the molecule relative to the active site. All the derivatives fluoresce in the solution and the crystalline states. Fluorescence characterization has focused on determination of binding effects on the probe quantum yield, lifetime, absorption and emission spectra, and quenching by added solutes. Quenching studies show that, as postulated, the Asp(sup 101)-bound probes are partially sheltered from the bulk solution by their location within the active site cleft. Probes bound to His(sup 15) have quenching constants about equal to those for the free probes, indicating that this site is highly exposed to the bulk solution.

Sumida, John P.↗

Mutations in the putative calcium-binding domain of polyomavirus VP1 affect capsid assembly

Calcium ions appear to play a major role in maintaining the structural integrity of the polyomavirus and are likely involved in the processes of viral uncoating and assembly. Previous studies demonstrated that a VP1 fragment extending from Pro-232 to Asp-364 has calcium-binding capabilities. This fragment contains an amino acid stretch from Asp-266 to Glu-277 which is quite similar in sequence to the amino acids that make up the calcium-binding EF hand structures found in many proteins. To assess the contribution of this domain to polyomavirus structural integrity, the effects of mutations in this region were examined by transfecting mutated viral DNA into susceptible cells. Immunofluorescence studies indicated that although viral protein synthesis occurred normally, infective viral progeny were not produced in cells transfected with polyomavirus genomes encoding either a VP1 molecule lacking amino acids Thr-262 through Gly-276 or a VP1 molecule containing a mutation of Asp-266 to Ala. VP1 molecules containing the deletion mutation were unable to bind 45Ca in an in vitro assay. Upon expression in Escherichia coli and purification by immunoaffinity chromatography, wild-type VP1 was isolated as pentameric, capsomere-like structures which could be induced to form capsid-like structures upon addition of CaCl2, consistent with previous studies. However, although VP1 containing the point mutation was isolated as pentamers which were indistinguishable from wild-type VP1 pentamers, addition of CaCl2 did not result in their assembly into capsid-like structures. Immunogold labeling and electron microscopy studies of transfected mammalian cells provided in vivo evidence that a mutation in this region affects the process of viral assembly.

Non-NASA Center↗

Earth Observations to Assess Impact of Hurricane Katrina on John C. Stennis Space Center

The peril from hurricanes to Space Operations Centers is real and is forecast to continue; Katrina, Rita, and Wilma of 2005 and Charley, Frances, Ivan, and Jeanne of 2004 are sufficient motivation for NASA to develop a multi-Center plan for preparedness and response. As was demonstrated at SSC (Stennis Space Center) in response to Hurricane Katrina, NASA Centers are efficiently activated as local command centers, playing host to Federal and State agencies and first responders to coordinate and provide evacuation, relocation, response, and recovery activities. Remote sensing decision support provides critical insight for managing NASA infrastructure and for assisting Center decision makers. Managers require geospatial information to manage the federal city. Immediately following Katrina, SSC s power and network connections were disabled, hardware was inoperative, technical staff was displaced and/or out of contact, and graphical decision support tools were non-existent or less than fully effective. Despite this circumstance, SSC EOC (Emergency Operations Center) implemented response operations to assess damage and to activate recovery plans. To assist Center Managers, the NASA ASP (Applied Sciences Program) made its archive of high-resolution data over the site available. In the weeks and months after the immediate crisis, NASA supplemented this data with high-resolution, post-Katrina imagery over SSC and much of the affected coastal areas. Much of the high-resolution imagery was made available through the Department of Defense Clear View contract and was distributed through U.S. Geological Survey Center for Earth Resources Observation and Science "Hurricane Katrina Disaster Response" Web site. By integrating multiple image data types with other information sources, ASP applied an all-source solutions approach to develop decision support tools that enabled managers to respond to critical issues, such as expedient access to infrastructure and deployment of resources, provision of temporary shelter, logistical control of critical supplies, and the mobilization and coordination of assets from ground crews to aircraft/airspace management. Furthermore, ASP developed information products that illustrate risks to SSC's infrastructure from surge, inundation, and flood. Current plans include developing wind-risk prototype products for refinement and adoption into EOC plans.

Graham, William D.↗

Airspace Systems Program: Next Generation Air Transportation System, NextGen Systems Analysis, Integration and Evaluation Project: Project Plan - Version 1.0

The key objectives of the NASA ASP are to: Improve mobility, capacity efficiency and access of the airspace system. Improve collaboration, predictability, and flexibility for the airspace users. Enable accurate modeling and simulation of air transportation systems. Accommodate operations of all classes of aircraft. Maintain system safety and environmental protection. In support of these program objectives, the major goal of the NextGen-SAIE Project is to enable the transition of key capacity and efficiency improvements to the NAS. Since many aspects of the NAS are unique to specific airport or airspace environments, demand on various parts of the NAS is not expected to increase equally as system demand grows. SAIE will provide systems level analysis of the NAS characteristics, constraints, and demands such that a suite of capacity-increasing concepts and technologies for system solutions are enabled and facilitated. The technical objectives in support of this goal are the following: Integration, evaluation, and transition of more mature concepts and technologies in an environment that faithfully emulates real-world complexities. Interoperability research and analysis of ASP technologies across ATM functions is performed to facilitate integration and take ASP concepts and technologies to higher Technology Readiness Level (TRL). Analyses are conducted on the program s concepts to identify the system benefits or impacts. System level analysis is conducted to increase understanding of the characteristics and constraints of airspace system and its domains.

Quon, Leighton↗

In Situ Analysis of Organics with a Portable Mass Spectrometer

The search for extra-terrestrial life starts at home. In order to find life on other planets, we start by examining life processes we understand on the earth. Though it may not be possible to see the life in the form of macroscopic organisms, telltale signs of life can exist in the form of small organic molecules such as peptides and amino acids. Our overall goal is to test a portable mass spectrometer (MS) system, the Mini 10.5, for astrobiological applications including in situ hydrocarbon analysis and sediments analysis using an additional automated sample processing system (ASPS). The collaborative research focuses on two current projects in the field of astrobiology. Both projects are geared towards examining organics distributed in extreme environments. One portion of study attempts to qualitatively analyze the effect of volatile organic compounds (VOC) produced by diesel exhaust on lichens growing in the desert. This requires measurements to be taken by bringing the instrument to the Mojave Desert and monitoring atmospheric composition of VOCs in situ. The second project is to evaluate the miniature MS system as a detector for the ASPS extraction system. A major obstacle of any chemometric in situ analysis is the suppression of analyte signal by concomitant signal from the surrounding environment. The ASPS extraction device has been developed at JPL to extract amino acids from sediment samples and elute them in solution. The solution is eluted at a high pH and needs to be conditioned to a more neutral pH so that dissolved amino acids can be readily protonated and subsequently analyzed by electrospray MS.

astrobiology↗

Thermal Design Challenges Posed by the Four Bed CO2 Scrubber COTS Air-Save Pump

The Four Bed Carbon Dioxide (4BCO2) scrubber Air-Save Pump (ASP) operates as part of the adsorbent bed regeneration cycle. The ASP removes residual air from the bed for return to the cabin prior to heat and vacuum exposure which drives out the CO2 regenerating the bed. 4BCO2 employs a Commercial Off-the-Shelf (COTS) scroll type air pump, repackaged in an acoustically insulated enclosure to reduce noise and mounted to a cold plate. The International Space Station (ISS) Low Temperature Loop (LTL) flow (operating between 38°F and 50°F) first cools the process air, and then flows through the cold plate, cooling the pump. This results in competing ASP thermal design goals: (1) to keep the pump and motor sufficiently cool and (2) to avoid forming condensation due to over-cooling. Surfaces below 60°F typically warrant careful consideration of condensation. A test-calibrated thermal model demonstrates such a balanced design is feasible with temperatures above 60°F. A separate, coupled fluid model predicts the potential for condensation formation, allowing risk assessment of flying with the unmodified design.

Daniel P Popok↗

Assessing the Needs of NASA's Near Real-Time Earth Observation Products

"The 2017-2027 Decadal Survey for Earth Science and Applications from Space stated that NASA's Earth Science with planned implementation of applications provides sustained earth observations for societal benefits [1]. The Decadal Survey indicated that data latency is invaluable for time-sensitive applications including disaster risk reduction, wildland fire carbon emissions quantification, real-time measurements of the state of the hydrologic systems and many more. Data latency refers to the time between earth observation and data products available to users. During the past 13 years, NASA's Land, Atmosphere Near Real-Time Capability for Earth Observing Systems (LANCE) continues to provide free access to earth observation products that are made available much quicker than routine processing allows. The latency of most LANCE data products is Near Real-time (NRT) which is defined as less than three hours from satellite observations [2]. LANCE is managed by the Earth Science Data and Information System (ESDIS) Project at NASA Goddard Space Flight Center [3], and a User Working Group (UWG) is responsible for providing guidance to LANCE. LANCE data are used by direct users and brokers who add value to the data [4]. NASA Earth Applied Sciences Program (ASP) is one of the primary users of LANCE, which collaborates with partner organizations and provides support to scientists to solve problems in applications of earth observations. ASP promotes the use of LANCE NRT data products to demonstrate applications in decision making, facilitates end-user feedback to the science team to improve data products, and provides information on future demands for research. LANCE supports applications that need a rapid response including detecting wildland fires and volcanic eruptions, tracking smoke, ash and dust plumes, monitoring air quality and tracking extreme weather events such as hurricanes, landslides, and floods. To gather feedback regarding the availability, accessibility and actionability of NASA's NRT data products for societal benefit, three surveys and a few discussions with experts involved in the topic within ASP were conducted from the perspective of users. Feedback has been collected from users who are interested in using low latency NASA data within application communities of agriculture, disasters, water resources, health and air quality, ecological conservation, wildland fires and capacity building. Analysis-ready NRT data products in a variety of formats have been mentioned many times in the collected feedback, especially for applied users with little to no experience using research-grade earth observation products. Users prefer to have products that can be easily integrated into their existing workflows and take their analysis to the data. HDF5 is a commonly used data format for research, but typically requires some conversion to a more friendly format for applications and regular use in decision-making. Users prefer the GeoTIFF data format that can be directly ingested into a GIS mapping software and platform for data analysis and visualization. For example, LANCE’s fire, flood, SO2 and Black Marble Nighttime Blue/Yellow Composite data products have been integrated into NASA Disasters Mapping Portal, which is an GIS-based open data portal, for users in the disaster management community. There are 291 LANCE NRT layers available through GIBS and Worldview, where users can download a snapshot in GeoTIFF format. Operational users expect data to be processed as close to the user as possible. The collected feedback indicates that LANCE fire products within 3 hours latency would meet the needs of the wildland fire community. The ideal latency for volcanic application is 10-15 minutes. Users in Volcanic Ash Advisory Centers (VAAC) reported that the first forecast volcanic product should be issued within 75 minutes from the volcano eruption [5]. Overall, for disaster applications, data latency within 3 hours is useful while latency greater than 12 hours is not timely enough for operational use. Capacity building and training are critical for users to be able to access, interpret and use data products and tools for their decision making, especially for applied users with limited experience using earth observation products. LANCE data products have been used in a number of capacity building projects domestically and internationally [6]. As LANCE continues to bring new products into the system, users request training to utilize LANCE new and upcoming data products and capabilities in their applications. Due to the limitation of bandwidth and downstream flow paths, users in some developing countries need tools to select and download data for a specific area of interest instead of bulk downloads. The collected feedback also shows the lack of available SAR satellite low latency data products. The advantages of SAR to monitor conditions and changes on the ground through darkness, clouds, volcanic ash, and other atmospheric conditions, are appealing to low latency users. For example, terabytes of low latency but cloudy optical images are not helpful in rapidly identifying the extent of flood or fire impacts. LANCE could be complemented with low latency measurements via the upcoming NASA-ISRO Synthetic Aperture Radar (NISAR) mission [7]. Requests for higher spatial resolution products are expressed. A user from the wildland fire management community reported that products with 30-m spatial resolution could be used to detect small fires. The 30-m Landsat OLI fire data is now part of NASA’s Fire Information for Resource Management System (FIRMS) US/Canada [8]. Within the open and free NASA resources, LANCE disseminates NRT data products in a manner that allows them to be accessible and understandable to both scientific and applied users. In many application areas, latency plays an important or even decisive role where low latency earth observations help people to observe areas of interest, detect and track changes in the environment and make timely decisions. NASA’s Earth Applied Sciences Program promotes the use of LANCE NRT products and builds a bridge between application users and research teams. The collected feedback indicates data latency within 3 hours is useful for most of the applications, and shows the needs of user-friendly, analysis-ready products, and requests training on LANCE’s new and upcoming data products. User feedback has been provided to LANCE UWG for guidance and recommendations, and for translating findings into something actionable.

Tian Yao↗

The study and development of an active scattering particle size spectrometer for space environments

The work is reported on laser operation in a vacuum, and the theoretical response of active scattering particle spectrometers (ASPS). The theoretical considerations connected with the thermal, interferometric and electrical problems that influence laser operation under vacuum conditions are discussed along with the scattering signal pulses with high frequency ripple content which were observed while operating the ASPS.

Knollenberg, R. G.↗

Digital controller design: Analysis of the annular suspension pointing system

The annular suspension and pointing system (ASPS) a payload auxiliary pointing device of the space shuttle is briefly described along with the function of the digital controller. The equations of motion of a simplified plan planar model of the ASPS are derived. Results of computer simulations are discussed.

Source record↗

The 1.1 micrometer and visible emission semiconductor diode lasers

In (AlGa)As, the first of three alloy systems studied, Continuous Wave (CW) operation was obtained at room temperature at a wavelength as low as 7260 A. Reliability in this system was studied in the incoherent mode. Zinc doped devices had significant degradation, whereas Ge or Ge plus Zi doped devices had none. The Al2O3 facet coatings were shown to significantly reduce facet deterioration in all types of lasers, longer wavelength units of that type having accumulated (at the time of writing) 22,000 hours with little if any degradation. A CL study of thin (AlGa)As layers revealed micro fluctuation in composition. A macro-scale fluctuation was observed by electroreflectance. An experimental and theoretical study of the effect of stripe width on the threshold current was carried out. Emission below 7000 A was obtained in VPE grown Ga(AsP) (In,Ga)P with CW operation at 10 C. Lasers and LED's were made by LPE in (InGa) (AsP). Laser thresholds of 5 kA/cm2 were obtained, while LED efficiences were on the order of 2%. Incoherent life test over 6000 hours showed no degradation.

Ladany, I.↗

The Annular Suspension and Pointing System /ASPS/

The Annular Suspension and Pointing System (ASPS) may be attached to a carrier vehicle for orientation, mechanical isolation, and fine pointing purposes applicable to space experiments. It has subassemblies for both coarse and vernier pointing. A fourteen-degree-of-freedom simulation of the ASPS mounted on a Space Shuttle has yielded initial performance data. The simulation describes: the magnetic actuators, payload sensors, coarse gimbal assemblies, control algorithms, rigid body dynamic models of the payload and Shuttle, and a control system firing model.

Anderson, W. W.↗

The Annular Suspension and Pointing System

The Annular Suspension and Pointing System (ASPS) is a general purpose mount designed to provide orientation, mechanical isolation, and fine pointing for space experiments. The ASPS consists of two assemblies, the first being a set of two gimbals attached to a carrier spacecraft (e.g., Space Shuttle) and providing coarse pointing, and the second a magnetic vernier pointing and isolation assembly attached to the inner gimbal of the first assembly and providing fine pointing. Discussion of the evolution of this concept, required related technology, and data from analyses and simulations predicting pointing accuracies that allowed the specification of hardware design requirements is presented.

Anderson, W. W.↗

Digital controller design: Analysis of the annular suspension pointing system

The Annular Suspension and Pointing System (ASPS) is a payload auxiliary pointing device of the Space Shuttle. The ASPS is comprised of two major subassemblies, a vernier and a coarse pointing subsystem. The experiment is attached to a mounting plate/rim combination which is suspended on magnetic bearing/actuators (MBA) strategically located about the rim. Fine pointing is achieved by gimballing the plate/rim within the MBA gaps. Control about the experiment line-of-sight is obtained through the use of a non-contacting rim drive and positioning torquer. All sensors used to close the servo loops on the vernier system are noncontacting elements. Therefore, the experiment is a free-flyer constrained only by the magnetic forces generated by the control loops.

Kuo, B. C.↗

An evaluation of the Interactive Software Invocation System (ISIS) for software development applications

The Interactive Software Invocation System (ISIS), which allows a user to build, modify, control, and process a total flight software system without direct communications with the host computer, is described. This interactive data management system provides the user with a file manager, text editor, a tool invoker, and an Interactive Programming Language (IPL). The basic file design of ISIS is a five level hierarchical structure. The file manager controls this hierarchical file structure and permits the user to create, to save, to access, and to purge pages of information. The text editor is used to manipulate pages of text to be modified and the tool invoker allows the user to communicate with the host computer through a RUN file created by the user. The IPL is based on PASCAL and contains most of the statements found in a high-level programming language. In order to evaluate the effectiveness of the system as applied to a flight project, the collection of software components required to support the Annular Suspension and Pointing System (ASPS) flight project were integrated using ISIS. The ASPS software system and its integration into ISIS is described.

Noland, M. S.↗

Annular suspension and pointing system with controlled DC electromagnets

The Annular Suspension and Pointing System (ASPS) developed by the Flight System division of Sperry Corporation is a six-degree of freedom payload pointing system designed for use with the space shuttle. This magnetic suspension and pointing system provides precise controlled pointing in six-degrees of freedom, isolation of payload-carrier disturbances, and end mount controlled pointing. Those are great advantages over the traditional mechanical joints for space applications. In this design, we first analyzed the assumed model of the single degree ASPS bearing actuator and obtained the plant dynamics equations. By linearizing the plant dynamics equations, we designed the cascade and feedback compensators such that a stable and satisfied result was obtained. The specified feedback compensator was computer simulated with the nonlinearized plant dynamics equations. The results indicated that an unstable output occurred. In other words, the designed feedback compensator failed. The failure of the design is due to the Taylor's series expansion not converging.

Vu, Josephine Lynn↗

Design and Implementation of a Digital Controller for a Vibration Isolation and Vernier Pointing System

This paper discusses the recommissioning of the Annular Suspension and Pointing System (ASPS), originally developed in the mid 1970's for pointing and vibration isolation of space experiments. The hardware was developed for NASA Langley Research Center by Sperry Flight Systems (now Honeywell Satellite Systems), was delivered to NASA in 1983. Recently, the hardware was loaned to Old Dominion University (ODU). The ASPS includes coarse gimbal assemblies and a Vernier Pointing Assembly (VPA) that utilize magnetic suspension to provide noncontacting vibration isolation and vernier pointing of the payload. The VPA is the main focus of this research. At ODU, the system has been modified such that it can now be operated in a l-g environment without a gravity offload. Suspension of the annular iron rotor in five degrees-of-freedom has been achieved with the use of modern switching power amplifiers and a digital controller implemented on a 486-class PC.

Neff, Daniel J.↗