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At least 523 records · Page 29

Integrin-like proteins are localized to plasma membrane fractions, not plastids, in Arabidopsis

Integrins are a large family of integral membrane proteins that function in signal transduction in animal systems. These proteins are conserved in vertebrates, invertebrates, and fungi. Evidence from previous research suggests that integrin-like proteins may be present in plants as well, and that these proteins may function in signal transduction during gravitropism. In past studies, researchers have used monoclonal and polyclonal antibodies to localize beta 1 integrin-like proteins in plants. However, there is a disparity between data collected from these studies, especially since molecular weights obtained from these investigations range from 55-120 kDa for integrin-like proteins. To date, a complete investigation which employs all three basic immunolabeling procedures, immunoblotting, immunofluorescence microscopy, and immunogold labeling, in addition to extensive fractionation and exhaustive controls, has been lacking. In this paper, we demonstrate that use of a polyclonal antibody against the cytoplasmic domain of avian beta 1-integrin can produce potential artifacts in immunolocalization studies. However, these problems can be eliminated through use of starchless mutants or proper specimen preparation prior to electrophoresis. We also show that this antibody, when applied within the described parameters and with careful controls, identifies a large (100 kDa) integrin-like protein that is localized to plasma membrane fractions in Arabidopsis.

NASA Discipline Plant Biology↗

Immunocytochemical localization of latent transforming growth factor-beta1 activation by stimulated macrophages

Transforming growth factor-beta1 (TGF-beta) is secreted in a latent form consisting of mature TGF-beta noncovalently associated with its amino-terminal propeptide, which is called latency associated peptide (LAP). Biological activity depends upon the release of TGF-beta from the latent complex following extracellular activation, which appears to be the key regulatory mechanism controlling TGF-beta action. We have identified two events associated with latent TGF-beta (LTGF-beta) activation in vivo: increased immunoreactivity of certain antibodies that specifically detect TGF-beta concomitant with decreased immunoreactivity of antibodies to LAP. Macrophages stimulated in vitro with interferon-gamma and lipopolysaccharide reportedly activate LTGF-beta via cell membrane-bound protease activity. We show through dual immunostaining of paraformaldehyde-fixed macrophages that such physiological TGF-beta activation is accompanied by a loss of LAP immunoreactivity with concomitant revelation of TGF-beta epitopes. The induction of TGF-beta immunoreactivity colocalized with immunoreactive betaglycan/RIII in activated macrophages, suggesting that LTGF-beta activation occurs on the cell surface. Confocal microscopy of metabolically active macrophages incubated with antibodies to TGF-beta and betaglycan/RIII prior to fixation supported the localization of activation to the cell surface. The ability to specifically detect and localize LTGF-beta activation provides an important tool for studies of its regulation.

NASA Discipline Radiation Health↗

Localization of the ANG II type 2 receptor in the microcirculation of skeletal muscle

Only functional studies have suggested the presence of the ANG II type 2 (AT2) receptor in the microcirculation. To determine the distribution of this receptor in the rat skeletal muscle microcirculation, a polyclonal rabbit anti-rat antiserum was developed and used for immunohistochemistry and Western blot analysis. The antiserum was prepared against a highly specific and antigenic AT2-receptor synthetic peptide and was validated by competition and sensitivity assays. Western blot analysis demonstrated a prominent, single band at approximately 40 kDa in cremaster and soleus muscle. Immunohistochemical analysis revealed a wide distribution of AT2 receptors throughout the skeletal muscle microcirculation in large and small microvessels. Microanatomic studies displayed an endothelial localization of the AT2 receptor, whereas dual labeling with smooth muscle alpha-actin also showed colocalization of the AT2 receptor with vascular smooth muscle cells. Other cells associated with the microvessels also stained positive for AT2 receptors. Briefly, this study confirms previous functional data and localizes the AT2 receptor to the microcirculation. These studies demonstrate that the AT2 receptor is present on a variety of vascular cell types and that it is situated in a fashion that would allow it to directly oppose ANG II type 1 receptor actions.

NASA Discipline Cardiopulmonary↗

Relation of systemic and local muscle exercise capacity to skeletal muscle characteristics in men with congestive heart failure

OBJECTIVES. The present study was undertaken to further characterize changes in skeletal muscle morphology and histochemistry in congestive heart failure and to determine the relation of these changes to abnormalities of systemic and local muscle exercise capacity. BACKGROUND. Abnormalities of skeletal muscle appear to play a role in the limitation of exercise capacity in congestive heart failure, but information on the changes in muscle morphology and biochemistry and their relation to alterations in muscle function is limited. METHODS. Eighteen men with predominantly mild to moderate congestive heart failure (mean +/- SEM New York Heart Association functional class 2.6 +/- 0.2, ejection fraction 24 +/- 2%) and eight age- and gender-matched sedentary control subjects underwent measurements of peak systemic oxygen consumption (VO2) during cycle ergometry, resistance to fatigue of the quadriceps femoris muscle group and biopsy of the vastus lateralis muscle. RESULTS. Peak VO2 and resistance to fatigue were lower in the patients with heart failure than in control subjects (15.7 +/- 1.2 vs. 25.1 +/- 1.5 ml/min-kg and 63 +/- 2% vs. 85 +/- 3%, respectively, both p < 0.001). Patients had a lower proportion of slow twitch, type I fibers than did control subjects (36 +/- 3% vs. 46 +/- 5%, p = 0.048) and a higher proportion of fast twitch, type IIab fibers (18 +/- 3% vs. 7 +/- 2%, p = 0.004). Fiber cross-sectional area was smaller, and single-fiber succinate dehydrogenase activity, a mitochondrial oxidative marker, was lower in patients (both p < or = 0.034). Likewise, the ratio of average fast twitch to slow twitch fiber cross-sectional area was lower in patients (0.780 +/- 0.06 vs. 1.05 +/- 0.08, p = 0.019). Peak VO2 was strongly related to integrated succinate dehydrogenase activity in patients (r = 0.896, p = 0.001). Peak VO2, resistance to fatigue and strength also correlated significantly with several measures of fiber size, especially of fast twitch fibers, in patients. None of the skeletal muscle characteristics examined correlated with exercise capacity in control subjects. CONCLUSIONS. These results indicate that congestive heart failure is associated with changes in the characteristics of skeletal muscle and local as well as systemic exercise performance. There are fewer slow twitch fibers, smaller fast twitch fibers and lower succinate dehydrogenase activity. The latter finding suggests that mitochondrial content of muscle is reduced in heart failure and that impaired aerobic-oxidative capacity may play a role in the limitation of systemic exercise capacity.

Non-NASA Center↗

Localization of the chaperone binding site

The hypothesis derived from models of the multi-oligomeric chaperone complex suggests that partially denatured proteins bind in a central cavity in the aggregate. To test this hypothesis, the molecular chaperone, alpha crystallin, was bound to partially denatured forms of gamma crystallin, and the binding site was visualized by immunogold localization. In an alternative approach, gold particles were directly complexed with gamma crystallin, followed by binding to the alpha crystallin aggregate. In both cases, binding was localized to the central region of the aggregate, confirming for the first time that partially denatured proteins do indeed bind to a central region of the molecular chaperone aggregate.

Non-NASA Center↗

AMP deaminase histochemical activity and immunofluorescent isozyme localization in rat skeletal muscle

The cellular distribution of AMP deaminase (AMPda) isozymes was documented for rat soleus and plantaris muscles, utilizing immunofluorescence microscopy and immunoprecipitation methods. AMPda is a ubiquitous enzyme existing as three distinct isozymes, A, B and C, which were initially purified from skeletal muscle, liver (and kidney), and heart, respectively. AMPda-A is primarily concentrated subsarcolemmally and intermyofibrillarly within muscle cells, while isozymes B and C are concentrated within non-myofiber elements of muscle tissue. AMPda-B is principally associated with connective tissues surrounding neural elements and the muscle spindle capsule, and AMPda-C is predominantly associated with circulatory elements, such as arterial and venous walls, capillary endothelium, and red blood cells. These specific localizations, combined with documented differences in kinetic properties, suggest multiple functional roles for the AMPda isozymes or temporal segregation of similar AMPda functions. Linkage of the AMPda substrate with adenosine production pathways at the AMP level and the localization of isozyme-C in vascular tissue suggest a regulatory role in the microcirculation.

Non-NASA Center↗

On the Formation of Shock Waves in Subsonic Flows With Local Supersonic Velocities

In the flow about a body with large subsonic velocity if the velocity of the approaching flow is sufficiently large, regions of local supersonic velocities are formed about the body. It is known from experiment that these regions downstream of the flow are always bounded by shock waves; a continuous transition of the supersonic velocity to the subsonic under the conditions indicated has never been observed. A similar phenomenon occurs in pipes. If at two cross sections of the pipe the velocity is subsonic and between these sections regions of local supersonic velocity are formed without completely occupying a single cross section, these regions are always bounded by shock waves.

Frankl, F. I.↗

Local and Total Density Measurements in Ice Shapes

Preliminary measurements of local and total densities inside ice shapes were obtained from ice shapes grown in the NASA Glenn Research Tunnel for a range of glaze ice, rime ice, and mixed phase ice conditions on a NACA 0012 airfoil at 0 angle of attack. The ice shapes were removed from the airfoil and a slice of ice 3 mm thick was obtained using a microtome. The resulting samples were then x-rayed to obtain a micro-radiography, the film was digitized, and image processing techniques were used to extract the local and total density values.

Vargas, Mario↗

NASA Structural Analysis Report on the American Airlines Flight 587 Accident - Local Analysis of the Right Rear Lug

A detailed finite element analysis of the right rear lug of the American Airlines Flight 587 - Airbus A300-600R was performed as part of the National Transportation Safety Board s failure investigation of the accident that occurred on November 12, 2001. The loads experienced by the right rear lug are evaluated using global models of the vertical tail, local models near the right rear lug, and a global-local analysis procedure. The right rear lug was analyzed using two modeling approaches. In the first approach, solid-shell type modeling is used, and in the second approach, layered-shell type modeling is used. The solid-shell and the layered-shell modeling approaches were used in progressive failure analyses (PFA) to determine the load, mode, and location of failure in the right rear lug under loading representative of an Airbus certification test conducted in 1985 (the 1985-certification test). Both analyses were in excellent agreement with each other on the predicted failure loads, failure mode, and location of failure. The solid-shell type modeling was then used to analyze both a subcomponent test conducted by Airbus in 2003 (the 2003-subcomponent test) and the accident condition. Excellent agreement was observed between the analyses and the observed failures in both cases. From the analyses conducted and presented in this paper, the following conclusions were drawn. The moment, Mx (moment about the fuselage longitudinal axis), has significant effect on the failure load of the lugs. Higher absolute values of Mx give lower failure loads. The predicted load, mode, and location of the failure of the 1985-certification test, 2003-subcomponent test, and the accident condition are in very good agreement. This agreement suggests that the 1985-certification and 2003- subcomponent tests represent the accident condition accurately. The failure mode of the right rear lug for the 1985-certification test, 2003-subcomponent test, and the accident load case is identified as a cleavage-type failure. For the accident case, the predicted failure load for the right rear lug from the PFA is greater than 1.98 times the limit load of the lugs. I.

Raju, Ivatury S↗

The cold land processes experiment (CLPX) local scale observatin site (LSOS)

The Local Scale Observation Site (LSOS) is the smallest study site of the Cold LandProcesses Experiment (CLPX) and is located within the Fraser Meso-cell Study Area (MSA), near the Fraser Experimental Forest Headquarters Facility, in Fraser, CO USA.The 100-m x 100-m site consists of a small open field, a managed dense canopy and an open, mixed age canopy. Unlike the other components of the experiment, which focus on spatial distributions at relatively brief snapshots in time, measurements at the local scale site focused on the temporal domain.

cold land processes ground-based remote sensing va↗

The Milky Way, the Local Group & the IR Tully-Fisher Diagram

Using the near infrared fluxes of local group galaxies derived from Cosmic Background Explorer/Diffuse Infrared Background Experiment band maps and published Cepheid distances, we construct Tully-Fisher diagrams for the Local Group.

Milky Way↗

Initiation of the Slow-Rise and Fast-Rise Phases of an Erupting Solar Filament by Localized Emerging Magnetic Field via Microflaring

EUV data from EIT show that a filament of 2001 February 28 underwent a slow-rise phase lasting about 6 hrs, before rapidly erupting in a fast-rise phase. Concurrent images in soft X-rays (SXRs) from Yohkoh/SXT show that a series of three microflares, prominent in SXT images but weak in EIT approx.195 Ang EUV images, occurred near one end of the filament. The first and last microflares occurred respectively in conjunction with the start of the slow-rise phase and the start of the fast-rise phase, and the second microflare corresponded to a kink in the filament trajectory. Beginning within 10 hours of the start of the slow rise, new magnetic flux emerged at the location of the microflaring. This localized new flux emergence and the resulting microflares, consistent with reconnection between the emerging field and the sheared sigmoid core magnetic field holding the filament, apparently caused the slow rise of this field and the transition to explosive eruption. For the first time in such detail, the observations show this direct action of localized emerging flux in the progressive destabilization of a sheared core field in the onset of a coronal mass ejection (CME). Similar processes may have occurred in other recently-studied events, NASA supported this work through NASA SR&T and SEC GI grants.

Sterling, A. C.↗

Local Neutral Density and Plasma Parameter Measurements in a Hollow Cathode Plume

In order to understand the cathode and keeper wear observed during the Extended Life Test (ELT) of the DS1 flight spare NSTAR thruster and provide benchmarking data for a 2D cathode/cathode-plume model, a basic understanding of the plasma and neutral gas parameters in the cathode orifice and keeper region of the cathode plume must be obtained. The JPL cathode facility is instrumented with an array of Langmuir probe diagnostics along with an optical diagnostic to measure line intensity of xenon neutrals. In order to make direct comparisons with the present model, a flat plate anode arrangement was installed for these tests. Neutral density is deduced from the scanning probe data of the plasma parameters and the measured xenon line intensity in the optical regime. The Langmuir probes are scanned both axially, out to 7.0 cm downstream of the keeper, and radially to obtain 2D profile of the plasma parameters. The optical fiber is housed in a collimating stainless steel tube, and is scanned to view across the cathode plume along cuts in front of the keeper with a resolution of 1.5 mm. The radial intensities are unfolded using the Abel inversion technique that produces radial profiles of local neutral density. In this paper, detailed measurements of the plasma parameters and the local neutral densities will be presented in the cathode/keeper plume region for a 1.5 cm diameter NEXIS cathode at 25A of discharge current at several different strengths of applied magnetic field.

Plasma density↗

Local Elastic Constants for Epoxy-Nanotube Composites from Molecular Dynamics Simulation

A method from molecular dynamics simulation is developed for determining local elastic constants of an epoxy/nanotube composite. The local values of C11, C33, K12, and K13 elastic constants are calculated for an epoxy/nanotube composite as a function of radial distance from the nanotube. While the results possess a significant amount of statistical uncertainty resulting from both the numerical analysis and the molecular fluctuations during the simulation, the following observations can be made. If the size of the region around the nanotube is increased from shells of 1 to 6 in thickness, then the scatter in the data reduces enough to observe trends. All the elastic constants determined are at a minimum 20 from the center of the nanotube. The C11, C33, and K12 follow similar trends as a function of radial distance from the nanotube. The K13 decreases greater distances from the nanotube and becomes negative which may be a symptom of the statistical averaging.

Frankland, S. J. V.↗

Method of remotely estimating a rest or best lock frequency of a local station receiver using telemetry

A system includes a remote station and a local station having a receiver. The receiver operates in an unlocked state corresponding to its best lock frequency (BLF). The local station derives data indicative of a ratio of the BLF to a reference frequency of the receiver, and telemeters the data to the remote station. The remote station estimates the BLF based on (i) the telemetered data, and (ii) a predetermined estimate of the reference frequency.

Fielhauer, Karl B.↗

An Analysis of Spacecraft Localization from Descent Image Data for Pinpoint Landing on Mars and other Cratered Bodies Data Acquisition

A pinpoint landing capability will be a critical component for many planned NASA missions to Mars and beyond. Implicit in the requirement is the ability to accurately localize the spacecraft with respect to the terrain during descent. In this paper, we present evidence that a vision-based solution using craters as landmarks is both practical and will meet the requirements of next generation missions. Our emphasis in this paper is on the feasibility of such a system in terms of (a) localization accuracy and (b) applicability to Martian terrain. We show that accuracy of well under 100 meters can be expected under suitable conditions. We also present a sensitivity analysis that makes an explicit connection between input data and robustness of our pose estimate. In addition, we present an analysis of the susceptibility of our technique to inherently ambiguous configurations of craters. We show that probability of failure due to such ambiguity is becoming increasingly small.

landforms↗

The Effects of Modeled Microgravity on Nucleocytoplasmic Localization of Human Apurinic/Apyrimidinic

Exposure to space radiation and microgravity occurs to humans during space flight. In order to have accurate risk estimations, answering questions to whether increased DNA damage seen during space flight in modified by microgravity are important. Several studies have examined whether intercellular repair of radiation-induced DNA lesions are modified by microgravity. Results from these studies show no modification of the repair processes due to microgravity. However, it is known that in studies not involving radiation that microgravity interferes with normal development. Interestingly, there is no data that attempts to analyze the possible effects of microgravity on the trafficking of DNA repair proteins. In this study, we analyze the effects of modeled microgravity on nucleocytoplasmic shuttling of the human DNA repair enzyme apurinic/apyrimidinic endonuclease 1 (APE1/Ref1) which is involved in base excision repair. We examined nuclear translocation of APE1 using enhanced green fluorescent protein (EGFP) fused to APE1 as a reporter. While APE1 under normal gravity showed normal nuclear localization, APE1 nuclear localization under modeled microgravity was decreased. These results suggest that nucleocytoplasmic translocation of APE1 is modified under modeled microgravity.

Gonda, Steve↗