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At least 1,171 records · Page 65

Characterization of Human Bone Alkaline Phosphatase in Pichia Pastoris

A soluble form of human bone alkaline phosphatase has been expressed in a recombinant strain of the methylotrophic yeast Pichia pastoris. We constructed a plasmid containing cDNA encoding for human bone alkaline phosphatase, with the hydrophobic carboxyl terminal portion deleted. Alkaline phosphatase was secreted into the medium to a level of 32mg/L when cultured in shake flasks, and enzyme activity was 12U/mg, as measured by a spectrophotometric assay. By conversion to a fermentation system, a yield of 880mg/L has been achieved with an enzyme activity of 968U/mg. By gel electrophoresis analysis, it appears that greater than 50% of the total protein in the fermentation media is alkaline phosphatase. Although purification procedures are not yet completely optimized, they are expected to include filtration, ion exchange and affinity chromatography. Our presentation will focus on the purification and crystallization results up to the time of the conference. Structural data should provide additional information on the role of alkaline phosphatase in normal bone mineralization and in certain bone mineralization anomalies.

Malone, Christine C.↗

Simple Refractometers for Index Measurements by Minimum Deviation Method from Far-ultraviolet to Near Infrared

The focal shift of an optical filter used in non-collimated light depends directly on substrate thickness and index of refraction. The HST Advanced Camera for Surveys (ACS) requires a set of filters whose focal shifts are tightly matched. Knowing the index of refraction for substrate glasses allows precise substrate thicknesses to be specified. Two refractometers have been developed at the Goddard Space Flight Center (GSFC) to determine the indices of refraction of materials from which ACS filters are made. Modem imaging detectors for the near infrared, visible, and far ultraviolet spectral regions make these simple yet sophisticated refractometers possible. A new technology, high accuracy, angular encoder also developed at GSFC makes high precision index measurement possible in the vacuum ultraviolet.

Leviton, Douglas B.↗

Situated Control Rules

The plan net formalism presented in this paper is a language for expressing possible actions independent of any particular plan. Plan nets encode non-deterministic control programs which specify the local conditions under which actions may be taken. A two stage analysis synthesizes situated control rules which act to reduce this non-deterministic choice such that all remaining alternatives for action necessarily satisfy any given goals. Action execution can precede situated control rule synthesis: situated control rules simply help ensure that actions are first steps on the way to eventual goal satisfaction. Situated control rules inform action; they do not define it.

Drummond, Mark↗

Statecharts Via Process Algebra

Statecharts is a visual language for specifying the behavior of reactive systems. The Language extends finite-state machines with concepts of hierarchy, concurrency, and priority. Despite its popularity as a design notation for embedded system, precisely defining its semantics has proved extremely challenging. In this paper, a simple process algebra, called Statecharts Process Language (SPL), is presented, which is expressive enough for encoding Statecharts in a structure-preserving and semantic preserving manner. It is establish that the behavioral relation bisimulation, when applied to SPL, preserves Statecharts semantics

Luttgen, Gerald↗

Position Estimation Verification Testing for the Video Guidance Sensor and Dynamic Overhead Target Simulator

The Video Guidance Sensor, part of the Automated Rendezvous and Capture mechanism, is due to undergo formal qualification testing at Marshall Space Flight Center. Before it undergoes this qualification, a test was needed to verify repeatability of the sensor, and to allow different sensor configurations to be compared. This test was developed at the Flight Robotics Laboratory. The test uses a software script to drive the sensor target to the same position and thus allows sensor runs to be compared. The sensor target is the Dynamic Overhead Target Simulator. The simulator uses encoders as its position indicator. Distance Measuring Device's were used to independently verify the software script, the sensor reading, and the target position. The test area, sensor, and other test equipment are briefly described. The actual data is tabulated and will serve as a baseline for future tests. The software script was found to be adequate for the test. Position repeatability was acceptable for all the equipment. The system test is now ready to be used in formal qualification testing.

Gaines, Joseph↗

"Interactive Classification Technology"

The investigators are upgrading a knowledge representation language called SL (Symbolic Language) and an automated reasoning system called SMS (Symbolic Manipulation System) to enable the technologies to be used in automated reasoning and interactive classification systems. The overall goals of the project are: a) the enhancement of the representation language SL to accommodate multiple perspectives and a wider range of meaning; b) the development of a sufficient set of operators to enable the interpreter of SL to handle representations of basic cognitive acts; and c) the development of a default inference scheme to operate over SL notation as it is encoded. As to particular goals the first-year work plan focused on inferencing and.representation issues, including: 1) the development of higher level cognitive/ classification functions and conceptual models for use in inferencing and decision making; 2) the specification of a more detailed scheme of defaults and the enrichment of SL notation to accommodate the scheme; and 3) the adoption of additional perspectives for inferencing.

deBessonet, Cary↗

Myosin Heavy Chain Gene Expression in Developing Neonatal Skeletal Muscle: Involvement of the Nerve, Gravity, and Thyroid State

The myosin heavy chain (MHC) gene family encodes at least six MHC proteins (herein designated as neonatal, embryonic, slow type I (beta), and fast IIa, IIx, and IIb) that are expressed in skeletal muscle in a muscle-specific and developmentally-regulated fashion. At birth, both antigravity (e.g. soleus) and locomotor (e.g., plantaris) skeletal muscles are undifferentiated relative to the adult MHC phenotype such that the neonatal and embryonic MHC isoforms account for 80 - 90% of the MHC pool in a fast locomotor muscle; whereas, the embryonic and slow, type I isoforms account for approx. 90% of the pool in a typical antigravity muscle. The goal of this study was to investigate the role of an intact nerve, gravity and thyroid hormone (T3), as well as certain interactions of these interventions, on MHC gene expression in developing neonatal skeletal muscles of rodents.

Baldwin, K. M.↗

Molecular Mechanisms Regulating Muscle Fiber Composition Under Microgravity

The overall goal of this project is to reveal the molecular mechanisms underlying the selective and debilitating atrophy of specific skeletal muscle fiber types that accompanies sustained conditions of microgravity. Since little is currently known about the regulation of fiber-specific gene expression programs in mammalian muscle, elucidation of the basic mechanisms of fiber diversification is a necessary prerequisite to the generation of therapeutic strategies for attenuation of muscle atrophy on earth or in space. Vertebrate skeletal muscle development involves the fusion of undifferentiated mononucleated myoblasts to form multinucleated myofibers, with a concomitant activation of muscle-specific genes encoding proteins that form the force-generating contractile apparatus. The regulatory circuitry controlling skeletal muscle gene expression has been well studied in a number of vertebrate animal systems. The goal of this project has been to achieve a similar level of understanding of the mechanisms underlying the further specification of muscles into different fiber types, and the role played by innervation and physical activity in the maintenance and adaptation of different fiber phenotypes into adulthood. Our recent research on the genetic basis of fiber specificity has focused on the emergence of mature fiber types and have implicated a group of transcriptional regulatory proteins, known as E proteins, in the control of fiber specificity. The restriction of E proteins to selected muscle fiber types is an attractive hypothetical mechanism for the generation of muscle fiber-specific patterns of gene expression. To date our results support a model wherein different E proteins are selectively expressed in muscle cells to determine fiber-restricted gene expression. These studies are a first step to define the molecular mechanisms responsible for the shifts in fiber type under conditions of microgravity, and to determine the potential importance of E proteins as upstream targets for the effects of weightlessness. In the past year we have determined that the expression of E Proteins is restricted to specific fiber types by post-transcriptional mechanisms. By far, the most prevalent mechanism of cellular control for achieving post-transcriptional regulation of gene expression is selective proteolysis -through the ubiquitin -proteasome pathway. Steady-state levels of HEB message are similar in all fast and slow skeletal muscle fiber types, yet the protein is restricted to Type IIX fibers. HEB appears to be a nodal point for regulating fiber-specific transcription, as expression of the transcription factor is regulated at the post-transcriptional level. It is not clear at present whether the regulation is at the level of protein synthesis or degradation. We are now poised to evaluate the biological role of ubiquitination in fiber specific-gene expression by controlling the post-transcriptional expression of E Proteins. The use of metabolic labelling and pharmacological inhibitors of the ubiquitin pathway will be used to identify the mode of regulation of the Type IIX expression pattern. The potential role of specific kinases in effecting the restriction of HEB expression will be examined by using both inhibitors and activators. The results of these studies will provide the necessary information to evaluate the biological role of E proteins in controlling fiber type transitions, and in potentially attenuating the atrophic effects of microgravity conditions. We have also recently shown that ectopic expression of the HEB protein transactivates the Type IIX-specific skeletal a-actin reporter. The 218 bp skeletal a-actin promoter drives transgene expression solely in mature Type IIX fibers. A mouse also carrying the transgene MLCI/HEB (which ectopically expresses the E Protein HEB in Type IIB fibers) forces expression of the skeletal a-actin reporter gene in Type IIB fibers. We can now dissect the composition of this fiber-specific cis-element. The skeletal a-actin promoter is quite compact and has been extensively characterized in vitro for activity and binding factors. The single E box may act as a binding target of myogenic factor/HEB heterodimer to allow for IIX expression. The HEB transcription factor may recognize either the precise flanking sequences of the E Box, or perhaps interacting with other proteins bound nearby, and activating expression in Type IIX fibers. This E box will be both ablated, and alternatively, as ablation may well destroy any muscle-specific transcriptional activity, flanking sequences substituted with those surrounding the E box (El) of the myogenin promoter. Modification of fiber-specific transgene expression will be tested in transgenic mice. The results of these studies will provide basic information on the regulatory circuitry underlying fiber specificity, and will form the basis for building appropriate transgenic regulatory cassettes to effect fiber transitions in subsequent experimental manipulations on unweighted muscles.

Rosenthal, Nadia A.↗

Improved Speech Coding Based on Open-Loop Parameter Estimation

A nonlinear optimization algorithm for linear predictive speech coding was developed early that not only optimizes the linear model coefficients for the open loop predictor, but does the optimization including the effects of quantization of the transmitted residual. It also simultaneously optimizes the quantization levels used for each speech segment. In this paper, we present an improved method for initialization of this nonlinear algorithm, and demonstrate substantial improvements in performance. In addition, the new procedure produces monotonically improving speech quality with increasing numbers of bits used in the transmitted error residual. Examples of speech encoding and decoding are given for 8 speech segments and signal to noise levels as high as 47 dB are produced. As in typical linear predictive coding, the optimization is done on the open loop speech analysis model. Here we demonstrate that minimizing the error of the closed loop speech reconstruction, instead of the simpler open loop optimization, is likely to produce negligible improvement in speech quality. The examples suggest that the algorithm here is close to giving the best performance obtainable from a linear model, for the chosen order with the chosen number of bits for the codebook.

Juang, Jer-Nan↗

Iterative Repair Planning for Spacecraft Operations Using the Aspen System

This paper describes the Automated Scheduling and Planning Environment (ASPEN). ASPEN encodes complex spacecraft knowledge of operability constraints, flight rules, spacecraft hardware, science experiments and operations procedures to allow for automated generation of low level spacecraft sequences. Using a technique called iterative repair, ASPEN classifies constraint violations (i.e., conflicts) and attempts to repair each by performing a planning or scheduling operation. It must reason about which conflict to resolve first and what repair method to try for the given conflict. ASPEN is currently being utilized in the development of automated planner/scheduler systems for several spacecraft, including the UFO-1 naval communications satellite and the Citizen Explorer (CX1) satellite, as well as for planetary rover operations and antenna ground systems automation. This paper focuses on the algorithm and search strategies employed by ASPEN to resolve spacecraft operations constraints, as well as the data structures for representing these constraints.

Rabideau, G.↗

First-Generation Jet Propulsion Laboratory "Hockey-Puck" Free-Flying Magnetometers for Distributed In-Situ Multiprobe Measurement of Current Density Filamentation in the Northern Auroral Zone: Enstrophy Mission

The sub-orbital rocket mission was a collaborative project between the University of New Hampshire, Cornell University, and the Jet Propulsion Laboratory (JPL) to study filamentation phenomena in the northern Auroral zone. The Enstrophy mission test flies the JPL Free-Flying Magnetometer (FFM) concept. The FFM technology development task has been funded by NASA develop miniaturized, low-power, integrated "sensorcrafts". JPL's role was to design, integrate, test, and deliver four FFMs for deployment from the sounding rocket, allowing a unique determination of curl-B. This provides a direct measurement of magnetic-field-aligned current density along the rocket trajectory. A miniaturized three-axis fluxgate magnetometer was integrated with a 4-channel 22-bit sigma-delta Analog to Digital Converter (ADC), four temperature sensors, digital control electronics, seven (Li-SOCl2) batteries, two (4 deg x 170 deg field of view) sun-sensors, a fan-shaped-beam laser diode beacon, a (16 MHz) stable Temperature Compensated Crystal Oscillator (TCXO) clock, Radio Frequency (RF) communication subsystem, and an antenna for approximately 15 minutes of operation where data was collected continuously and transmitted in three (3) bursts (approximately 26 seconds each) to ground station antennas at Poker Flat, Alaska. FFMs were stowed within two trays onboard the rocket during the rocket launch and were released simultaneously using the spinning action of the rocket at approximately 300 km altitude (approximately 100 sec. into the flight). FFMs were deployed with spin rate of approximately 17 Hz and approximately 3 m/sec linear velocity with respect to the rocket. For testing purposes while the rocket was in the launch pad and during flight prior to release of FFMs from the rocket, commands (such as "power on", "test", "flight", "power off', and clock "Reset" signal) were transmitted via a infrared Light Emitting Diode to an infrared detector in the FFM. Special attention was paid to low magnetic signature electronic design and choice of materials in packaging. The miniaturized fluxgate magnetometers had a range of 1-60000 nT with 0.1% full-scale linearity. The frequency range of interest for magnetic measurement was 10 mHz - 50 Hz. Digital data from the magnetometer's three axes were placed in a 4MB Static Random Access Memory (SRAM) in data packages (frames) formatted together with time tags and frame ID. After a specified time was elapsed, the data were Viterbi encoded and transmitted at a rate of 100 kbps (BPSK). Each of the four FFMs transmitted at different frequency. These carrier frequencies were in the range of 2200-2300 MHz. The antenna was a single patch on a high dielectric constant substrate covering one end-plate of the hockey-puck-sized unit. The local clocks aboard the FFMs were reset at the start of the mission and stayed synchronized within 3 msec during the mission. Position of each FFM with respect to the rocket is calculated by the knowledge of its release velocity (measured at exit point of the FFM launcher tract) providing an accuracy of 1 m over the maximum range of 3 km. Spatial and temporal nature of observants can be separated to within 3 m in space or 3 msec time interval.

Javadi, H.↗

A Real-Time High Performance Data Compression Technique For Space Applications

A high performance lossy data compression technique is currently being developed for space science applications under the requirement of high-speed push-broom scanning. The technique is also error-resilient in that error propagation is contained within a few scan lines. The algorithm is based on block-transform combined with bit-plane encoding; this combination results in an embedded bit string with exactly the desirable compression rate. The lossy coder is described. The compression scheme performs well on a suite of test images typical of images from spacecraft instruments. Hardware implementations are in development; a functional chip set is expected by the end of 2001.

Yeh, Pen-Shu↗

Information-Theoretic Assessment of Sample Imaging Systems

By rigorously extending modern communication theory to the assessment of sampled imaging systems, we develop the formulations that are required to optimize the performance of these systems within the critical constraints of image gathering, data transmission, and image display. The goal of this optimization is to produce images with the best possible visual quality for the wide range of statistical properties of the radiance field of natural scenes that one normally encounters. Extensive computational results are presented to assess the performance of sampled imaging systems in terms of information rate, theoretical minimum data rate, and fidelity. Comparisons of this assessment with perceptual and measurable performance demonstrate that (1) the information rate that a sampled imaging system conveys from the captured radiance field to the observer is closely correlated with the fidelity, sharpness and clarity with which the observed images can be restored and (2) the associated theoretical minimum data rate is closely correlated with the lowest data rate with which the acquired signal can be encoded for efficient transmission.

Huck, Friedrich O.↗

Free-Flying Magnetometer Data System

The Free-Flying Magnetometer (FFM) is an autonomous "sensorcraft" developed at the Jet Propulsion Laboratory (JPL) for the Enstrophy sounding rocket mission. This mission was a collaborative project between the University of New Hampshire, Cornell University and JPL. The science goal of the mission was the study of current filamentation phenomena in the northern auroral region through multipoint measurements of magnetic field. The technical objective of the mission was the proof of concept of the JPL FFM design and the demonstration of an in-situ multipoint measurement technique employing many free-flying spacecraft. Four FFMs were successfully deployed from a sounding rocket launched from Poker Flats, Alaska on February 11, 1999. These hockey-puck-sized (80 mm diameter, 38 mm. height, 250 gram mass) free flyers each carry a miniature 3-axis flux-gate magnetometer that output +/- 2 V signals corresponding to a +/- 60,000 nT measurement range for each axis. The FFM uses a synchronized four-channel Sigma(Delta) Analog-to-Digital Converter (ADC) having a dynamic range of +/- 2.5V and converting at a rate of 279 samples/second/channel. Three channels are used to digitize the magnetometer signals to 17-bit (1.144 nT/bit) resolution. The fourth ADC channel is multiplexed for system monitoring of four temperature sensors and two battery voltages. The FFM also contains two sun sensors, a laser diode which emits a fan-shaped beam, a miniature S-band transmitter for direct communication to the ground station antennas, an ultra-stable Temperature Compensated Crystal Oscillator (TCXO) clock, an integrated data subsystem implemented in a Field-Programmable Gate Array (FPGA), a 4 Mbit Static Random Access Memory (SRAM) for data storage and Lithium Thionyl Chloride batteries for power. Communicating commands to the FFM prior to deployment is achieved with an infrared (IR) link. The FFM IR receiver responds to 9-bit pulse coded signals that are generated by an IR Light Emitting Diode (LED) in the payload for turning FFM power on or off and placing the FFM in a test mode or flight mode. The IR links are also used to synchronize (zero) the clocks onboard all the FFMs through a reset pulse originating from the payload GPS receiver that is issued when the FFMs are in flight mode. The FPGA based data subsystem manages continuous data collection from the four ADC channels and sun sensors, formatting and storing the data to SRAM, and controlling downlink transmission. The transmitter is powered only after a 2547 frame SRAM buffer has been filled (approx. 5 minutes of data). The data is Viterbi encoded and sent to the S-band transmitter via a First-In-First-Out (FIFO) buffer who's output is clocked at 100 bits/second. After the 26-second transmission, the transmitter is turned off to reduce noise coupling to the sensitive magnetometer. The data subsystem control consists of a master state machine that performs data flow management and is interfaced through a prioritized interrupt scheme to state machines that service the ADC, sun sensors and transmitter FIFO. Continuous data collection prevents the missing of data during transmission and provides implicit time tagging of the data acquired by the ADC because of synchronization with the TCXO clock.

Blaes, B.↗

A Wideband Satcom Based Avionics Network with CDMA Uplink and TDM Downlink

The purpose of this paper is to describe some key technical ideas behind our vision of a future satcom based digital communication network for avionics applications The key features of our design are as follows: (a) Packetized transmission to permit efficient use of system resources for multimedia traffic; (b) A time division multiplexed (TDM) satellite downlink whose physical layer is designed to operate the satellite link at maximum power efficiency. We show how powerful turbo codes (invented originally for linear modulation) can be used with nonlinear constant envelope modulation, thus permitting the satellite amplifier to operate in a power efficient nonlinear regime; (c) A code division multiple access (CDMA) satellite uplink, which permits efficient access to the satellite from multiple asynchronous users. Closed loop power control is difficult for bursty packetized traffic, especially given the large round trip delay to the satellite. We show how adaptive interference suppression techniques can be used to deal with the ensuing near-far problem; (d) Joint source-channel coding techniques are required both at the physical and the data transport layer to optimize the end-to-end performance. We describe a novel approach to multiple description image encoding at the data transport layer in this paper.

Agrawal, D.↗

Visually Lossless Data Compression for Real-Time Frame/Pushbroom Space Science Imagers

A visually lossless data compression technique is currently being developed for space science applications under the requirement of high-speed push-broom scanning. The technique is also applicable to frame based imaging and is error-resilient in that error propagation is contained within a few scan lines. The algorithm is based on a block transform of a hybrid of modulated lapped transform (MLT) and discrete cosine transform (DCT), or a 2-dimensional lapped transform, followed by bit-plane encoding; this combination results in an embedded bit string with exactly the desirable compression rate as desired by the user. The approach requires no unique table to maximize its performance. The compression scheme performs well on a suite of test images typical of images from spacecraft instruments. Flight qualified hardware implementations are in development; a functional chip set is expected by the end of 2001. The chip set is being designed to compress data in excess of 20 Msamples/sec and support quantizations from 2 to 16 bits.

Yeh, Pen-Shu↗

Uniform Data Access Using GXD

This paper gives an overview of GXD, a framework facilitating publication and use of data from diverse data sources. GXD defines an object-oriented data model designed to represent a wide range of things including data, its metadata, resources and query results. GXD also defines a data transport language. a dialect of XML, for representing instances of the data model. This language allows for a wide range of data source implementations by supporting both the direct incorporation of data and the specification of data by various rules. The GXD software library, proto-typed in Java, includes client and server runtimes. The server runtime facilitates the generation of entities containing data encoded in the GXD transport language. The GXD client runtime interprets these entities (potentially from many data sources) to create an illusion of a globally interconnected data space, one that is independent of data source location and implementation.

Vanderbilt, Peter↗

Overexpression of Human Bone Alkaline Phosphatase in Pichia Pastoris

The Pichiapastoris expression system was utilized to produce functionally active human bone alkaline phosphatase in gram quantities. Bone alkaline phosphatase is a key enzyme in bone formation and biomineralization, yet important questions about its structural chemistry and interactions with other cellular enzymes in mineralizing tissues remain unanswered. A soluble form of human bone alkaline phosphatase was constructed by deletion of the 25 amino acid hydrophobic C-terminal region of the encoding cDNA and inserted into the X-33 Pichiapastoris strain. An overexpression system was developed in shake flasks and converted to large-scale fermentation. Alkaline phosphatase was secreted into the medium to a level of 32mgAL when cultured in shake flasks. Enzyme activity was 12U/mg measured by a spectrophotometric assay. Fermentation yielded 880mgAL with enzymatic activity of 968U/mg. Gel electrophoresis analysis indicates that greater than 50% of the total protein in the fermentation is alkaline phosphatase. A purification scheme has been developed using ammonium sulfate precipitation followed by hydrophobic interaction chromatography. We are currently screening crystallization conditions of the purified recombinant protein for subsequent X-ray diffraction analyses. Structural data should provide additional information on the role of alkaline phosphatase in normal bone mineralization and in certain bone mineralization anomalies.

Karr, Laurel↗