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At least 199 records · Page 11

Eosinophilia of dystrophin-deficient muscle is promoted by perforin-mediated cytotoxicity by T cell effectors

Previous investigations have shown that cytotoxic T lymphocytes (CTLs) contribute to muscle pathology in the dystrophin-null mutant mouse (mdx) model of Duchenne muscular dystrophy through perforin-dependent and perforin-independent mechanisms. We have assessed whether the CTL-mediated pathology includes the promotion of eosinophilia in dystrophic muscle, and thereby provides a secondary mechanism through which CTLs contribute to muscular dystrophy. Quantitative immunohistochemistry confirmed that eosinophilia is a component of the mdx dystrophy. In addition, electron microscopic observations show that eosinophils traverse the basement membrane of mdx muscle fibers and display sites of close apposition of eosinophil and muscle membranes. The close membrane apposition is characterized by impingement of eosinophilic rods of major basic protein into the muscle cell membrane. Transfer of mdx splenocytes and mdx muscle extracts to irradiated C57 mice by intraperitoneal injection resulted in muscle eosinophilia in the recipient mice. Double-mutant mice lacking dystrophin and perforin showed less eosinophilia than was displayed by mdx mice that expressed perforin. Finally, administration of prednisolone, which has been shown previously to reduce the concentration of CTLs in dystrophic muscle, produced a significant reduction in eosinophilia. These findings indicate that eosinophilia is a component of the mdx pathology that is promoted by perforin-dependent cytotoxicity of effector T cells. However, some eosinophilia of mdx muscle is independent of perforin-mediated processes.

NASA Discipline Musculoskeletal↗

Power Distribution for Cryogenic Instruments at 6-40K The James Webb Space Telescope Case

The Integrated Science Instrument Module (ISIM) of the James Webb Space Telescope (JWST) operates its instruments passively cooled at around 40 Kelvin (K), with a warm Instrument Electronic Compartment (IEC) at 300K attached to it. From the warm electronics all secondary signal and power harnesses have to bridge this 300-40K temperature difference and minimize the power dissipation and parasitic heat leak into the cold region. After an introduction of the ISIM with its instruments, the IEC with the electronics, and the harness architecture with a special radiator, this paper elaborates on the cryogenic wire selection and tests performed to establish current de-rating rules for different wire types. Finally failure modes are analyzed for critical instrument interfaces that could inject excessive currents and heat into the harness and cold side, and several solutions for the removal of such failures are presented.

Rumler, Peter↗

Numerical modeling of multidimensional flow in seals and bearings used in rotating machinery

The rotordynamic behavior of turbomachinery is critically dependent on fluid dynamic rotor forces developed by various types of seals and bearings. The occurrence of self-excited vibrations often depends on the rotor speed and load. Misalignment and rotor wobbling motion associated with differential clearance were often attributed to stability problems. In general, the rotative character of the flowfield is a complex three dimensional system with secondary flow patterns that significantly alter the average fluid circumferential velocity. A multidimensional, nonorthogonal, body-fitted-grid fluid flow model is presented that describes the fluid dynamic forces and the secondary flow pattern development in seals and bearings. Several numerical experiments were carried out to demonstrate the characteristics of this complex flowfield. Analyses were performed by solving a conservation form of the three dimensional Navier-Stokes equations transformed to those for a rotating observer and using the general-purpose computer code PHOENICS with the assumptions that the rotor orbit is circular and that static eccentricity is zero. These assumptions have enabled a precise steady-state analysis to be used. Fluid injection from ports near the seal or bearing center increased fluid-film direct dynamic stiffness and, in some cases, significantly increased quadrature dynamic stiffness. Injection angle and velocity could be used for active rotordynamic control; for example, injection, when compared with no injection, increased direct dynamic stiffness, which is an important factor for hydrostatic bearings.

Hendricks, R. C.↗

Wavefront-Error Performance Characterization for the James Webb Space Telescope (JWST) Integrated Science Instrument Module (ISIM) Science Instruments

The science instruments (SIs) comprising the James Webb Space Telescope (JWST) Integrated Science Instrument Module (ISIM) were tested in three cryogenic-vacuum test campaigns in the NASA Goddard Space Flight Center (GSFC)'s Space Environment Simulator (SES) test chamber. In this paper, we describe the results of optical wavefront-error performance characterization of the SIs. The wavefront error is determined using image-based wavefront sensing, and the primary data used by this process are focus sweeps, a series of images recorded by the instrument under test in its as-used configuration, in which the focal plane is systematically changed from one image to the next. High-precision determination of the wavefront error also requires several sources of secondary data, including 1) spectrum, apodization, and wavefront-error characterization of the optical ground-support equipment (OGSE) illumination module, called the OTE Simulator (OSIM), 2) F-number and pupil-distortion measurements made using a pseudo-nonredundant mask (PNRM), and 3) pupil geometry predictions as a function of SI and field point, which are complicated because of a tricontagon-shaped outer perimeter and small holes that appear in the exit pupil due to the way that different light sources are injected into the optical path by the OGSE. One set of wavefront-error tests, for the coronagraphic channel of the Near-Infrared Camera (NIRCam) Longwave instruments, was performed using data from transverse translation diversity sweeps instead of focus sweeps, in which a sub-aperture is translated and/or rotated across the exit pupil of the system. Several optical-performance requirements that were verified during this ISIM-level testing are levied on the uncertainties of various wavefront-error-related quantities rather than on the wavefront errors themselves. This paper also describes the methodology, based on Monte Carlo simulations of the wavefront-sensing analysis of focus-sweep data, used to establish the uncertainties of the wavefront-error maps.

image analysis↗

Artemis I: Test Flight Buys Down Risk for Humanity's Return to the Moon

The Artemis program successfully demonstrated its foundational deep space human transportation system during the Artemis I test flight in late 2022. This bold mission put the Artemis program on a course to accomplish increasingly complex missions to return humans to the Moon and to prepare for Mars and other destinations. The Artemis I uncrewed mission tested the Space Launch System (SLS) rocket and the Orion spacecraft using the ground processing, launch and recovery services provided by the Exploration Ground Systems (EGS) program at the Kennedy Space Center (KSC). The Artemis team accomplished key mission priorities that deliberately bought down risk for upcoming crewed missions as part of a build-up flight test strategy. The Artemis I mission priorities were 1) demonstrate Orion heatshield at lunar re-entry conditions; 2) operate systems in flight environment; and 3) retrieve the spacecraft. The successful performance and demonstration of these primary mission objectives from lift-off thru orbital insertion, translunar injection, outbound transit towards the Moon, entry into lunar orbit, coast in the lunar Distant Retrograde Orbit (DRO), exit from lunar orbit, transit back to Earth, atmospheric re-entry, descent, splashdown and recovery successfully showed that NASA has a foundational deep space transportation capability in place for human class missions. In addition to the primary mission objectives, a set of scientific payloads, approved Flight Test Objectives (FTOs) and Development Flight Test Objectives (DFTOs) were incorporated into the nominal mission plan and were designated as secondary, or priority 4, and were considered “bonus objectives” in terms of crewed flight risk buydown. During the mission, the team found itself in a position where it was comfortable achieving even more content than originally planned. As a result, DFTOs were added during the mission to further buy down risk to later crewed test flights. This paper presents an overview of the Artemis I test flight, including mission priorities and key test objectives, the launch campaign and as-flown mission profile, test objectives accomplished over the course of the 25.5- day mission, and implications to the crewed Artemis II and later test flight missions.

Artemis↗

Flow visualization study of a vortex-wing interaction

A flow visualization study in water was completed on the interaction of a streamwise vortex with a laminar boundary layer on a two-dimensional wing. The vortex was generated at the tip of a finite wing at incidence, mounted perpendicular to the main wing, and having the same chord as the main wing. The Reynolds number based on wing chord was about 5000. Two different visualization techniques were used. One involved the injection of two different colored dyes into the vortex and the boundary layer. The other technique utilized hydrogen bubbles as an indicator. The position of the vortex was varied in a directional normal to the wing. The angle of attack of the main wing was varied from -5 to +12.5 deg. The vortex induced noticeable cross flows in the wing boundary layer from a distance equivalent to 0.75 chords. When very close to the wing, the vortex entrained boundary layer fluid and caused a cross flow separation which resulted in a secondary vortex.

Mehta, R. D.↗

SWIRL as a means of liquid management in low gravity

Swirling of a liquid in a container may prove to be a more desirable method of managing liquids in low gravity (space) environments than by rotating the entire container. By injecting a relatively high velocity liquid tangentially into the body of the fluid, swirl can best be started rapidly, however an estimate of the quantity and velocity of jetflow, or mechanical power of a pump impeller required to maintain a given radial acceleration (G force) is needed to assess the feasibility of such a method. While the key aspect of the problem is determining the rate of rotational energy dissipation by wall friction in the container, there are other considerations, and the present study investigates the possible additional effects of axial variation of tangential velocity and secondary (radial and axial) flow components within the rotating fluid.

Steward, W. Gene↗

Autoignition of hydrogen injected transverse to a supersonic airstream

A parametric study of autoignition characteristics of transverse fuel injector patterns for a hydrogen-fueled, supersonic-combustion ramjet was conducted in direct-connect tests over a range of simulated hypersonic flight conditions. Autoignition limits, defined directly by visual observations and indirectly by measured wall pressure and temperature, are compared with a semi-empirical ignition limit model which accounts for first order effects on autoignition. It is shown that to initiate combustion in the immediate region of the fuel injector, ignition must occur in the separation region ahead of the fuel jet. Autoignition within this separation zone exhibits strong dependence on flow pressure and temperature, injector diameter, local boundary layer energy thickness, wall temperature, and on fluid dynamic interactions with adjacent downstream facing steps (such as inlet isolation steps). Secondary effects are also discussed. The empirical model is used to define limits of test temperature, pressure, and scale required for autoignition in a current subscale engine test program.

Mcclinton, C. R.↗

Throttling Characteristics of the RL10 Derivative Common Extensible Cryogenic Engine -- Demo 1.6 and 1.7 Test Results

The Common Extensible Cryogenic Engine (CECE) is a deep throttling cryogenic Lunar Module Descent Engine (LMDE) technology development demonstrator based on the Pratt & Whitney Rocketdyne (PWR) RL10 engine. Previous testing on this engine occurred during 2006 as Demo 1.0, then Demo 1.5 in 2007, Demo 1.6 in 2008 and finally Demo 1.7 in 2010. A review of Demo 1.0, 1.5, and preliminary results of 1.6 were reported in previous JANNAF papers. Demo 1.6 was tested at the PWR E-6 test facility in November 2008. The primary goal of this series was to mitigate low frequency combustion instability observed at low power levels. To mitigate the chug, the Demo 1.6 injector was modified from the previous configurations to include an approximately 0.050 inch thick teflon-type spray-on insulation to reduce heat transfer to the LOX manifold, which was believed to be a significant contributor to the low power instability. In addition, gaseous helium injection into the LOX manifold was used as a means to stabilize the system. Also explored in this test series was mitigation for a low power 1 Hz fuel system oscillation caused by sub-critical hydrogen boiling in the chamber cooling jacket. Reduced area gas venturis were utilized to avoid the 1 Hz fuel-size oscillation by keeping the cooling jacket supercritical down to lower engine power levels. Demo 1.7 began testing in March 2010. Its primary objectives were to demonstrate closed loop control capability on mixture ratio and chamber pressure, start to lower power levels and increased throttling ramp rates. Secondary test objectives that are discussed include multiple engine starts, higher mixture ratio excursions and additional time at low power level. While the complete test series and data reduction is not yet complete for Demo 1.7, an overview and up-to-date status is provided.

Devine, Matthew K.↗

Active Fail-Safe Micro-Array Flow Control for Advanced Embedded Propulsion Systems

The primary objective of this research effort was to develop and analytically demonstrate enhanced first generation active "fail-safe" hybrid flow-control techniques to simultaneously manage the boundary layer on the vehicle fore-body and to control the secondary flow generated within modern serpentine or embedded inlet S-duct configurations. The enhanced first-generation technique focused on both micro-vanes and micro-ramps highly-integrated with micro -jets to provide nonlinear augmentation for the "strength' or effectiveness of highly-integrated flow control systems. The study focused on the micro -jet mass flow ratio (Wjet/Waip) range from 0.10 to 0.30 percent and jet total pressure ratios (Pjet/Po) from 1.0 to 3.0. The engine bleed airflow range under study represents about a 10 fold decrease in micro -jet airflow than previously required. Therefore, by pre-conditioning, or injecting a very small amount of high-pressure jet flow into the vortex generated by the micro-vane and/or micro-ramp, active flow control is achieved and substantial augmentation of the controlling flow is realized.

Anderson, Bernhard H.↗

Efferent projections of the ectostriatum in the pigeon (Columba livia)

The ectostriatum is a major visual component of the avian telencephalon. The core region of the ectostriatum (Ec) receives visual input from the optic tectum through thalamic nuclei. In the present study, the efferent projections of the ectostriatum were investigated by using the anterograde tracers Phaseolus vulgaris leucoagglutinin and biotinylated dextran amine. Projection patterns resulting from these tracers were confirmed by the retrograde tracer cholera toxin subunit B. When anterograde tracers were injected in Ec, primary projections were seen traveling dorsolaterally to the belt region of the ectostriatum (Ep) and the neostriatal area immediately surrounding Ep (Ep2). Neurons in Ep sent projections primarily to the overlying Ep2. The efferents of Ep2 traveled dorsolaterally to terminate in three telencephalic regions, from anterior to posterior: (1) neostriatum frontale, pars lateralis (NFL), (2) area temporo-parieto-occipitalis (TPO), and (3) neostriatum intermedium, pars lateralis (NIL). A part of the archistriatum intermedium and the lateral part of the neostriatum caudale also received somewhat minor projections. In addition, some neurons in Ec were also the source of direct, but minor, projections to the NFL, TPO, NIL, and archistriatum intermedium. The topographical relationship among the primary (Ec), secondary (Ep and Ep2), and tertiary (NFL, TPO, NIL) areas indicate that the neural populations for visual processing are organized along the rostral-caudal axis. Thus, the anterior Ec sent efferents to the anterior Ep, which in turn sent projections to anterior Ep2. Neurons in the anterior Ep2 sent projections to NFL and the anterior TPO. Similarly, the intermediate and posterior Ec sent projections to corresponding parts of Ep, whose efferents projected to intermediate and posterior Ep2, respectively. The intermediate Ep2 gave rise to major projections to TPO, whereas posterior Ep2 neurons sent efferents primarily to NIL. The organization of this neural circuit is compared with those of other sensory circuits in the avian telencephalon, as well as the laminar arrangement of the mammalian isocortex.

Non-NASA Center↗

Combustion Stability Characteristics of the Project Morpheus Liquid Oxygen/Liquid Methane Main Engine

The Project Morpheus liquid oxygen (LOX) / liquid methane rocket engines demonstrated acousticcoupled combustion instabilities during sea‐level ground‐based testing at the NASA Johnson Space Center (JSC) and Stennis Space Center (SSC). High‐amplitude, 1T, 1R, 1T1R (and higher order) modes appear to be triggered by injector conditions. The instability occurred during the Morpheus‐specific engine ignition/start sequence, and did demonstrate the capability to propagate into mainstage. However, the instability was never observed to initiate during mainstage, even at low power levels. The Morpheus main engine is a JSC‐designed ~5,000 lbf‐thrust, 4:1 throttling, pressure‐fed cryogenic engine using an impinging element injector design. Two different engine designs, named HD4 and HD5, and two different builds of the HD4 engine all demonstrated similar instability characteristics. Through the analysis of more than 200 hot fire tests on the Morpheus vehicle and SSC test stand, a relationship between ignition stability and injector/chamber pressure was developed. The instability has the distinct characteristic of initiating at high relative injection pressure drop (dP) at low chamber pressure (Pc); i.e., instabilities initiated at high dP/Pc at low Pc during the start sequence. The high dP/Pc during start results during the injector /chamber chill‐in, and is enhanced by hydraulic flip in the injector orifice elements. Because of the fixed mixture ratio of the existing engine design (the main valves share a common actuator), it is not currently possible to determine if LOX or methane injector dP/Pc were individual contributors (i.e., LOX and methane dP/Pc typically trend in the same direction within a given test). The instability demonstrated initiation characteristic of starting at or shortly after methane injector chillin. Colder methane (e.g., sub‐cooled) at the injector inlet prior to engine start was much more likely to result in an instability. A secondary effect of LOX sub‐cooling was also possibly observed; greater LOX sub‐ cooling improved stability. Some tests demonstrated a low‐amplitude 1L‐1T instability prior to LOX injector chill‐in. The Morpheus main engine also demonstrated chug instabilities during some engine shutdown sequences on the flight vehicle and SSC test stand. The chug instability was also infrequently observed during the startup sequence. The chug instabilities predictably initiated at low dP/Pc at low Pc. The chug instabilities were always self‐limiting; startup chug instabilities terminated during throttle‐up and shutdown chug instabilities decayed by shutdown termination.

Melcher, J. C.↗

Prompt Emission of GRB 121217A from Gamma-Rays to the Near-Infrared

The mechanism that causes the prompt-emission episode of gamma-ray bursts (GRBs) is still widely debated despite there being thousands of prompt detections. The favoured internal shock model relates this emission to synchrotron radiation. However, it does not always explain the spectral indices of the shape of the spectrum, which is often fit with empirical functions, such as the Band function. Multi-wavelength observations are therefore required to help investigate the possible underlying mechanisms that causes the prompt emission. We present GRB 121217A, for which we were able to observe its near-infrared (NIR) emission during a secondary prompt-emission episode with the Gamma-Ray burst Optical Near-infrared Detector (GROND) in combination with the Swift and Fermi satellites, which cover an energy range of 5 orders of magnitude (10(exp −3) keV to 100 keV). We determine a photometric redshift of z = 3.1 +/- 0.1 with a line-of-sight with little or no extinction (AV approx. 0 mag) utilising the optical/NIR SED. From the afterglow, we determine a bulk Lorentz factor of Gamma(sub 0) approx. 250 and an emission radius of R < 1018 cm. The prompt-emission broadband spectral energy distribution is well fit with a broken power law with beta1 = −0.3 +/- 0.1 and beta2 = 0.6 +/- 0.1 that has a break at E = 6.6 +/- 0.9 keV, which can be interpreted as the maximum injection frequency. Self-absorption by the electron population below energies of Ea < 6 keV suggest a magnetic field strength of B approx. 10(exp 5) G. However, all the best fit models underpredict the flux observed in the NIR wavelengths, which also only rebrightens by a factor of approx. 2 during the second prompt emission episode, in stark contrast to the X-ray emission, which rebrightens by a factor of approx. 100. This suggests an afterglow component is dominating the emission. We present GRB 121217A, one of the few GRBs that has multi-wavelength observations of the prompt-emission period and shows that it can be understood with a synchrotron radiation model. However, due to the complexity of the GRB's emission, other mechanisms that result in Band-like spectra cannot be ruled out.

prompt↗

Spray combustion under oscillatory pressure conditions

The performance and stability of liquid rocket engines is often argued to be significantly impacted by atomization and droplet vaporization processes. In particular, combustion instability phenomena may result from the interactions between the oscillating pressure field present in the rocket combustor and the fuel and oxidizer injection process. Few studies have been conducted to examine the effects of oscillating pressure fields on spray formation and its evolution under rocket engine conditions. The pressure study is intended to address the need for such studies. In particular, two potentially important phenomena are addressed in the present effort. The first involves the enhancement of the atomization process for a liquid jet subjected to an oscillating pressure field of known frequency and amplitude. The objective of this part of the study is to examine the coupling between the pressure field and or the resulting periodically perturbed velocity field on the breakup of the liquid jet. In particular, transverse mode oscillations are of interest since such modes are considered of primary importance in combustion instability phenomena. The second aspect of the project involves the effects of an oscillating pressure on droplet coagulation and secondary atomization. The objective of this study is to examine the conditions under which phenomena following the atomization process are affected by perturbations to the pressure or velocity fields. Both coagulation and represent a coupling mechanism between the pressure field and the energy release process in rocket combustors. It is precisely this coupling which drives combustion instability phenomena. Consequently, the present effort is intended to provide the fundamental insights needed to evaluate these processes as important mechanisms in liquid rocket instability phenomena.

Jacobs, H. R.↗

Effect of vane twist on the performance of dome swirlers for gas turbine airblast atomizers

For advanced gas turbine engines, two combustor systems, the lean premixed/prevaporized (LPP) and the rich burn/quick quench/lean burn (RQL) offer great potential for reducing NO(x) emissions. An important consideration for either concept is the development of an advanced fuel injection system that will provide a stable, efficient, and very uniform combustion system over a wide operating range. High-shear airblast fuel injectors for gas turbine combustors have exhibited superior atomization and mixing compared with pressure-atomizing fuel injectors. This improved mixing has lowered NO(x) emissions and the pattern factor, and has enabled combustors to alternate fuels while maintaining a stable, efficient combustion system. The performance of high-shear airblast fuel injectors is highly dependent on the design of the dome swirl vanes. The type of swirl vanes most widely used in gas turbine combustors are usually flat for ease of manufacture, but vanes with curvature will, in general, give superior aerodynamic performance. The design and performance of high-turning, low-loss curved dome swirl vanes with twist along the span are investigated. The twist induces a secondary vortex flow pattern which will improve the atomization of the fuel, thereby producing a more uniform fuel-air distribution. This uniform distribution will increase combustion efficiency while lowering NO(x) emissions. A systematic swirl vane design system is presented based on one-, two-, and three-dimensional flowfield calculations, with variations in vane-turning angle, rate of turning, vane solidity, and vane twist as design parameters.

Micklow, Gerald J.↗

Effect of vane twist on the performance of dome swirlers for gas turbine airblast atomizers

For advanced gas turbine engines, two combustor systems, the lean premixed/prevaporized (LPP) and the rich burn/quick quench/lean burn (RQL) offer great potential for reducing NO(x) emissions. An important consideration for either concept is the development of an advanced fuel injection system that will provide a stable, efficient, and very uniform combustion system over a wide operating range. High-shear airblast fuel injectors for gas turbine combustors have exhibited superior atomization and mixing compared with pressure-atomizing fuel injectors. This improved mixing has lowered NO(x) emissions and the pattern factor, and has enabled combustors to alternate fuels while maintaining a stable, efficient combustion system. The performance of high-shear airblast fuel injectors is highly dependent on the design of the dome swirl vanes. The type of swirl vanes most widely used in gas turbine combustors are usually flat for ease of manufacture, but vanes with curvature will, in general, give superior aerodynamic performance. The design and performance of high-turning, low-loss curved dome swirl vanes with twist along the span are investigated. The twist induces a secondary vortex flow pattern which will improve the atomization of the fuel, thereby producing a more uniform fuel-air distribution. This uniform distribution will increase combustion efficiency while lowering NO(x) emissions. A systematic swirl vane design system is presented based on one-, two-, and three-dimensional flowfield calculations, with variations in vane-turning angle, rate of turning, vane solidity, and vane twist as design parameters.

Micklow, Gerald J.↗

Three-dimensional Structure of Straight and Curved Plane Wakes

Although the plane wake is marked by the formation of strong spanwise vortices, the initially two-dimensional Karman-like vortices soon develop a three-dimensional structure in the form of secondary streamwise vortices. So far, this streamwise vortex structure has been studied mostly through flow visualization and at relatively low Reynolds numbers. The primary objective of the present program was to investigate the origin and evolution of the three-dimensional structure of straight and curved plane wakes at relatively high Reynolds numbers (Re(sub b) = 28,000) through detailed measurements of the mean and turbulent properties at several streamwise locations. The experiments were conducted in three phases. In the first phase, the development of a straight plane wake was investigated. In the second phase, the effects of imposed streamwise curvature on the wake development were examined. The streamwise curvature was of constant radius and very mild in terms of the curvature ratio (b/(square root of R) is less than 2 percent). In both the first and second phases, the role of initial conditions was examined in wakes generated from both untripped (laminar) and tripped (turbulent) initial boundany layers. In the third phase, the effects of injecting streamwise vorticity and the effects of increased Reynolds number on the tripped wake structure and development were investigated.

Weygandt, James H.↗

A provisional regulatory gene network for specification of endomesoderm in the sea urchin embryo

We present the current form of a provisional DNA sequence-based regulatory gene network that explains in outline how endomesodermal specification in the sea urchin embryo is controlled. The model of the network is in a continuous process of revision and growth as new genes are added and new experimental results become available; see http://www.its.caltech.edu/~mirsky/endomeso.htm (End-mes Gene Network Update) for the latest version. The network contains over 40 genes at present, many newly uncovered in the course of this work, and most encoding DNA-binding transcriptional regulatory factors. The architecture of the network was approached initially by construction of a logic model that integrated the extensive experimental evidence now available on endomesoderm specification. The internal linkages between genes in the network have been determined functionally, by measurement of the effects of regulatory perturbations on the expression of all relevant genes in the network. Five kinds of perturbation have been applied: (1) use of morpholino antisense oligonucleotides targeted to many of the key regulatory genes in the network; (2) transformation of other regulatory factors into dominant repressors by construction of Engrailed repressor domain fusions; (3) ectopic expression of given regulatory factors, from genetic expression constructs and from injected mRNAs; (4) blockade of the beta-catenin/Tcf pathway by introduction of mRNA encoding the intracellular domain of cadherin; and (5) blockade of the Notch signaling pathway by introduction of mRNA encoding the extracellular domain of the Notch receptor. The network model predicts the cis-regulatory inputs that link each gene into the network. Therefore, its architecture is testable by cis-regulatory analysis. Strongylocentrotus purpuratus and Lytechinus variegatus genomic BAC recombinants that include a large number of the genes in the network have been sequenced and annotated. Tests of the cis-regulatory predictions of the model are greatly facilitated by interspecific computational sequence comparison, which affords a rapid identification of likely cis-regulatory elements in advance of experimental analysis. The network specifies genomically encoded regulatory processes between early cleavage and gastrula stages. These control the specification of the micromere lineage and of the initial veg(2) endomesodermal domain; the blastula-stage separation of the central veg(2) mesodermal domain (i.e., the secondary mesenchyme progenitor field) from the peripheral veg(2) endodermal domain; the stabilization of specification state within these domains; and activation of some downstream differentiation genes. Each of the temporal-spatial phases of specification is represented in a subelement of the network model, that treats regulatory events within the relevant embryonic nuclei at particular stages. (c) 2002 Elsevier Science (USA).

Non-NASA Center↗