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At least 163 records · Page 9

Phosphorylation of the budgerigar fledgling disease virus major capsid protein VP1

The structural proteins of the budgerigar fledgling disease virus, the first known nonmammalian polyomavirus, were analyzed by isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The major capsid protein VP1 was found to be composed of at least five distinct species having isoelectric points ranging from pH 6.45 to 5.85. By analogy with the murine polyomavirus, these species apparently result from different modifications of an initial translation product. Primary chicken embryo cells were infected in the presence of 32Pi to determine whether the virus structural proteins were modified by phosphorylation. SDS-PAGE of the purified virus structural proteins demonstrated that VP1 (along with both minor capsid proteins) was phosphorylated. Two-dimensional analysis of the radiolabeled virus showed phosphorylation of only the two most acidic isoelectric species of VP1, indicating that this posttranslational modification contributes to VP1 species heterogeneity. Phosphoamino acid analysis of 32P-labeled VP1 revealed that phosphoserine is the only phosphoamino acid present in the VP1 protein.

Non-NASA Center↗

Eye-head coordination during optokinetic stimulation in squirrel monkeys

Head and eye movements in the yaw plane were recorded during and after optokinetic stimulation in squirrel monkeys. 1) Phasic or tonic head deviations to the side of the ocular quick phase occurred in 94% of total recordings (n = 50) during the perstimulus period, and in 75% of recordings (n = 49) during the poststimulus period. Magnitude of mean head deviation was significantly different between perstimulus and poststimulus periods. 2) Head nystagmus associated with eye nystagmus was consistently observed in seven of nine squirrel monkeys during optokinetic stimulation. Squirrel monkeys are thereby less prone to display head nystagmus than either guinea pigs, pigeons or chickens. 3) Slow phase speeds of coupled head and eye nystagmus were subjected to statistical analysis. A highly significant negative correlation was found between slow phase head and eye speeds. The correlation coefficient was - 0.81 at 60 degrees / sec stimulus (n = 119) and -0.72 at 100 degrees / sec stimulus (n = 131). The gaze speed, calculated by summing the head and eye speeds, was 59.1 plus or minus 6.8 / sec at 60 degrees / sec and 92.2 plus or minus 11.4 at 100 degrees / sec stimulus. There was no significant difference between the gaze speed in a free head condition and the eye speed when the head was fixed.

Eye Movements↗

Incidence of Latent Virus Shedding during Space Flight

Measurements of immune parameters of both cellular and innate immunity indicate alterations in immune function in astronauts. Immune changes are due to stress and perhaps other factors associated with launch, flight, and landing phases. Medical relevance of observed changes is not known. The reactivation of latent viruses has been identified as an important in vivo indicator of clinically relevant immune changes. The polymerase chain reaction (PCR) was used to detect the presence of specific viral DNA in body fluids. Initial studies demonstrated Epstein-Barr virus (EBV) reactivation during all 3 mission phases. EBV is shed in saliva following reactivation from B-cells. Incidence of EBV in saliva was higher than control subjects during all 3 mission phases. However, quantitative PCR revealed 10-fold higher levels of EBV DNA present in saliva collected during flight than found in pre- and post flight specimens. To determine if other latent viruses showed similar effects, cytomegalovirus (CMV), another herpes virus, shed in urine following reactivation was studied. A very low incidence (less than 2%) of CMV in urine is found in healthy, lowstressed individuals. However, 25-50% of astronauts shed CMV in their urine before, during, or after flight. Our studies are now focused on varicella-zoster virus (VZV), the etiological agent of chicken-pox during childhood and shingles later in life. We demonstrated reactivation of VZV and shedding of the virus during and after spaceflight in saliva of astronauts with no sign of active infection or symptoms. The maximum shedding of VZV occurred during the flight phase and diminishes rapidly during the first five days after landing. We have utilized the same PCR assay for VZV in a clinical study of shingles patients. Generally, shingles patients shed much more VZV in saliva than astronauts. However, the VZV levels in astronauts overlap with the lower range of VZV numbers in shingles patients. Saliva from shingles patients and astronauts were cultured and infectious VZV was recovered from both groups. We have concluded that multiple latent viruses do reactivate before, during, and after spaceflight and serve as very sensitive indicators for diminished cellular immunity. Future plans will be focused on the clinical risks posed by the reactivation of these viruses. Initial efforts will determine the effect of longer missions on the International Space Station on the reactivation patterns of these viruses.

Mehta, Satish K.↗

History of the Animal Care Program at Johnson Space Center

NASA has a rich history of scientific research that has been conducted throughout our numerous manned spaceflight programs. This scientific research has included animal test subjects participating in various spaceflight missions, including most recently, Space Shuttle mission STS-131. The Animal Care Program at Johnson Space Center (JSC) in Houston, Texas is multi-faceted and unique in scope compared to other centers within the agency. The animal care program at JSC has evolved from strictly research to include a Longhorn facility and the Houston Zoo's Attwater Prairie Chicken refuge, which is used to help repopulate this endangered species. JSC is home to more than 300 species of animals including home of hundreds of white-tailed deer that roam freely throughout the center which pose unique issues in regards to population control and safety of NASA workers, visitors and tourists. We will give a broad overview of our day to day operations, animal research, community outreach and protection of animals at NASA Johnson Space Center.

Khan-Mayberry, Noreen↗

Rapid Detection of the Varicella Zoster Virus in Saliva

Varicella zoster virus (VZV) causes chicken pox on first exposure (usually in children), and reactivates from latency causing shingles (usually in adults). Shingles can be extremely painful, causing nerve damage, organ damage, and blindness in some cases. The virus can be life-threatening in immune-compromised individuals. The virus is very difficult to culture for diagnosis, requiring a week or longer. This invention is a rapid test for VZV from a saliva sample and can be performed in a doctor s office. The kit is small, compact, and lightweight. Detec tion is sensitive, specific, and noninvasive (no needles); only a saliva sample is required. The test provides results in minutes. The entire test is performed in a closed system, with no exposure to infectious materials. The components are made mostly of inexpensive plastic injection molded parts, many of which can be purchased off the shelf and merely assembled. All biological waste is contained for fast, efficient disposal. This innovation was made possible because of discovery of a NASA scientists flight experiment showing the presence of VZV in saliva during high stress periods and disease. This finding enables clinicians to quickly screen patients for VZV and treat the ones that show positive results with antiviral medicines. This promotes a rapid recovery, easing of pain and symptoms, and reduces chances of complications from zoster. Screening of high-risk patients could be incorporated as part of a regular physical exam. These patients include the elderly, pregnant women, and immune-compromised individuals. In these patients, VZV can be a life-threatening disease. In both high- and low-risk patients, early detection and treatment with antiviral drugs can dramatically decrease or even eliminate the clinical manifestation of disease.

Pierson, Duane L.↗

Swarm Verification

Reportedly, supercomputer designer Seymour Cray once said that he would sooner use two strong oxen to plow a field than a thousand chickens. Although this is undoubtedly wise when it comes to plowing a field, it is not so clear for other types of tasks. Model checking problems are of the proverbial "search the needle in a haystack" type. Such problems can often be parallelized easily. Alas, none of the usual divide and conquer methods can be used to parallelize the working of a model checker. Given that it has become easier than ever to gain access to large numbers of computers to perform even routine tasks it is becoming more and more attractive to find alternate ways to use these resources to speed up model checking tasks. This paper describes one such method, called swarm verification.

grid computing↗

Design of Space Systems to Enable In-space Assembly and Servicing

For several decades, NASA has employed in-space systems to enhance the performance and extend the useful life of operational orbital assets. In at least one case, an operational mission was not only enhanced, but enabled – the International Space Station was made possible by crewed and robotic in-space assembly, and continues to support installation and operation of new science and technology payloads. In several cases (Hubble Space Telescope, Intelsat 401, Westar and Palapa), major operational assets were rescued or repaired soon after launch when otherwise mission-ending anomalies occurred or were detected. In addition to the original rescue, Hubble was upgraded four times, enabling high-demand, world class science over four decades. More recently, two Northrop Grumman Mission Extension Vehicles have captured two Intelsat spacecraft near the end of their life and fuel capacity, to take over maneuvering duties. In spite of these recent operational achievements, and with the exception of large human exploration vehicles and large space telescopes, space architects rarely consider in-orbit servicing and assembly capabilities in their future planning. Technologies such as multi-launch mission architectures (and rendezvous and proximity operations systems), docking systems, external robotics, advanced tools, modular systems and structures, and fluid transfer systems are available today to support these missions. In-space manufacturing will soon be operational to enable resilient missions that recover from on-orbit failures, and expand the utilization of space. We envision a future that includes these capabilities, and discuss the cultural, engineering, and technological challenges to achieving this vision. We discuss the vision, the proverbial chicken and the egg (which came first, the serviceable spacecraft or the servicer?), the cost, risk, and perceptions thereof of in-space operations, a “spectrum” of cooperative servicing design considerations, and the current status of the space industry’s slow but steady march to widespread operational use of on-orbit servicing, assembly, and manufacturing.

Bo Naasz↗

Impact Forces From Artificial and Real Birds on a Large Diameter Hopkinson Bar

Forces were measured from normal impacts with two different artificial bird recipes and two species of real birds on a very rigid flat surface. The tests were conducted by launching the soft body projectiles axially into a Hopkinson bar in accordance with the SAE AS6940 test standard at three different nominal impact conditions: (a) a 1 kg projectile at an impact velocity of 49 m/sec; (b) a 1.8 kg projectile at an impact velocity of 110 m/sec; and (c) a 1.8 kg projectile at an impact velocity of 310 m/sec. At each condition two artificial birds and one real bird projectile were tested with at least three test repeats. Simulations were conducted to assess the effects of projectile orientation and velocity on predicted forces. The Hopkinson bar consisted of an Aluminum 6061 solid cylindrical bar with a diameter of 305 mm and a length of 7.31 m, made up of two 3.66 m long sections that were in axial contact with each other. The bar was instrumented for strain measurement at two locations, 457.2 mm and 609.6 mm (1.5 and 2 diameters) from the impacted face. Forces were calculated from the measured strain. In addition, digital image correlation (DIC) was used to measure the velocity of the free end of the bar. The real bird projectiles were 1 kg Mallard ducks and 1.8 kg Golden Comet chickens that were prepared per ASTM F330-21. Prior to testing, the real birds were placed in muslin bags to minimize tumbling during flight. Two artificial bird formulations were tested, one produced by the University of Dayton Research Institute (UDRI) and the other by the German Aerospace Center, DLR. The UDRI projectile was a homogeneous, relatively soft solid, formulated with a mixture of water, gelatin and phenolic micro-balloons to achieve a nominal density of 0.95 g/cc. The DLR projectile consisted of a printed plastic shell with internal plastic ribs and filled with a gel material. The projectiles were designed such that the overall average density was the desired value of 0.95 g/cc. Initial testing showed that contact between the two Hopkinson bar segments was not sufficient to allow full transmission of the waves through the interface. For the 49 m/sec and 110 m/sec tests this resulted in reflections returning to the strain measurement site prior to the end of the impact. For these tests only a portion of the force pulse could be accurately measured. For the highest velocity tests at 310 m/sec the force pulse was complete by the time reflected waves interfered with the response. Despite this limitation, useful force data were obtained for all velocities. Impact forces were very sensitive to the impact orientation and velocity. The force history generally consisted of an initial region with varying amplitude followed by a relatively constant amplitude response. The steady state region was similar among all projectiles and similar to what would be predicted for an inviscid fluid with a density of 0.95 g/cc. The initial part of the force response exhibited varying degrees of test-to-test repeatability associated with each projectile type. Forces calculated from DIC velocity measurements of the free end of the bar compared favorably those from the strain measurements. In general, the artificial projectile impact forces were quite repeatable from test to test, with some exceptions which could be explained by issues associated with either the projectile itself or with how the projectile exited the gun barrel. For all impact velocities the initial portion of the impact force from the bird projectiles exhibited large test-to-test variability. In this paper, impact forces for three types of projectiles at three impact velocities will be presented with an emphasis on the test-to-test repeatability of the results. Based on test and simulation results a proposed method for normalizing the impact force to minimize effects of differences in projectile orientation, impact velocity and density will also be discussed.

Bird strike↗

Functional recovery of anterior semicircular canal afferents following hair cell regeneration in birds

Streptomycin sulfate (1.2 g/kg i.m.) was administered for 5 consecutive days to 5-7-day-old white Leghorn chicks; this causes damage to semicircular canal hair cells that ultimately regenerate to reform the sensory epithelium. During the recovery period, electrophysiological recordings were taken sequentially from anterior semicircular canal primary afferents using an indentation stimulus of the canal that has been shown to mimic rotational stimulation. Chicks were assigned to an early (14-18 days; n = 8), intermediate (28-34 days; n = 5), and late (38-58 days; n = 4) period based on days after treatment. Seven untreated chicks, 15-67 days old, provided control data. An absence of background and indent-induced discharge was the prominent feature of afferents in the early period: only "silent" afferents were encountered in 5/8 experiments. In several of these chicks, fascicles of afferent fibers were seen extending up to the epithelium that was void of hair cells, and intra- and extracellular biocytin labeling revealed afferent processes penetrating into the supporting cell layer of the crista. In 3/8 chicks 74 afferents could be characterized, and they significantly differed from controls (n = 130) by having a lower discharge rate and a negligible response to canal stimulation. In the intermediate period there was considerable variability in discharge properties of 121 afferents, but as a whole the number of "silent" fibers in the canal nerve diminished, the background rate increased, and a response to canal stimulation detected. Individually biocytin-labeled afferents had normal-appearing terminal specializations in the sensory epithelium by 28 days poststreptomycin. In the late period, afferents (n = 58) remained significantly different from controls in background discharge properties and response gain. The evidence suggests that a considerable amount of variability exists between chicks in the return of vestibular afferent function following ototoxic injury and that the secretory function of regenerating hair cells might become functional before their transducer function.

NASA Discipline Neuroscience↗

Inhibition of caspases prevents ototoxic and ongoing hair cell death

Sensory hair cells die after acoustic trauma or ototoxic insults, but the signal transduction pathways that mediate hair cell death are not known. Here we identify several important signaling events that regulate the death of vestibular hair cells. Chick utricles were cultured in media supplemented with the ototoxic antibiotic neomycin and selected pharmacological agents that influence signaling molecules in cell death pathways. Hair cells that were treated with neomycin exhibited classically defined apoptotic morphologies such as condensed nuclei and fragmented DNA. Inhibition of protein synthesis (via treatment with cycloheximide) increased hair cell survival after treatment with neomycin, suggesting that hair cell death requires de novo protein synthesis. Finally, the inhibition of caspases promoted hair cell survival after neomycin treatment. Sensory hair cells in avian vestibular organs also undergo continual cell death and replacement throughout mature life. It is unclear whether the loss of hair cells stimulates the proliferation of supporting cells or whether the production of new cells triggers the death of hair cells. We examined the effects of caspase inhibition on spontaneous hair cell death in the chick utricle. Caspase inhibitors reduced the amount of ongoing hair cell death and ongoing supporting cell proliferation in a dose-dependent manner. In isolated sensory epithelia, however, caspase inhibitors did not affect supporting cell proliferation directly. Our data indicate that ongoing hair cell death stimulates supporting cell proliferation in the mature utricle.

Non-NASA Center↗

Enhanced animal growth via ligand-regulated GHRH myogenic-injectable vectors

Regulated animal growth occurred following a single electroporated injection of a mixture of two plasmids (10 microg of DNA), one expressing the GeneSwitch regulator protein, the other an inducible growth hormone releasing hormone (GHRH) gene, into the tibialis anterior muscles of adult SCID mice. Administration of the ligand mifepristone (MFP) up-regulated GHRH expression, as shown by elevations of IGF-I levels, and when MFP dosing was withdrawn, IGF-I returned to baseline levels. Five cycles of IGF-I induction were observed during a five-month period. Chronic MFP dosing for 25 days increased lean body mass, weight gain, and bone mineral density significantly compared with non-MFP treated controls. In summary, long-term drug-regulated GHRH expression was achieved following plasmid-based gene therapy, and chronic induction of GHRH expression in adult animals led to improvements in weight gain and body composition.

NASA Discipline Musculoskeletal↗

Spectroscopic characterization of collagen cross-links in bone

Collagen is the most abundant protein of the organic matrix in mineralizing tissues. One of its most critical properties is its cross-linking pattern. The intermolecular cross-linking provides the fibrillar matrices with mechanical properties such as tensile strength and viscoelasticity. In this study, Fourier transform infrared (FTIR) spectroscopy and FTIR imaging (FTIRI) analyses were performed in a series of biochemically characterized samples including purified collagen cross-linked peptides, demineralized bovine bone collagen from animals of different ages, collagen from vitamin B6-deficient chick homogenized bone and their age- and sex-matched controls, and histologically stained thin sections from normal human iliac crest biopsy specimens. One region of the FTIR spectrum of particular interest (the amide I spectral region) was resolved into its underlying components. Of these components, the relative percent area ratio of two subbands at approximately 1660 cm(-1) and approximately 1690 cm(-1) was related to collagen cross-links that are abundant in mineralized tissues (i.e., pyridinoline [Pyr] and dehydrodihydroxylysinonorleucine [deH-DHLNL]). This study shows that it is feasible to monitor Pyr and DHLNL collagen cross-links spatial distribution in mineralized tissues. The spectroscopic parameter established in this study may be used in FTIRI analyses, thus enabling the calculation of relative Pyr/DHLNL amounts in thin (approximately 5 microm) calcified tissue sections with a spatial resolution of approximately 7 microm.

Non-NASA Center↗

Phase sensitive x-ray diffraction imaging of defects in biological macromolecular crystals

Conventional x-ray diffraction topography is currently used to map defects in the bulk of protein crystals, but the lack of sufficient contrast is frequently a limiting factor. We experimentally demonstrate that this barrier can be circumvented using a method that combines phase sensitive and diffraction imaging principles. Details of defects revealed in tetragonal lysozyme and cubic ferritin crystals are presented and discussed. The approach enabling the detection of the phase changes of diffracted x rays should prove to be useful in the study of defect structures in a broad range of biological macromolecular crystals.

Muramidase/chemistry↗