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At least 19 records

Replication-deficient adenovirus vector transfer of gfp reporter gene into supraoptic nucleus and subfornical organ neurons

The present studies used defined cells of the subfornical organ (SFO) and supraoptic nuclei (SON) as model systems to demonstrate the efficacy of replication-deficient adenovirus (Ad) encoding green fluorescent protein (GFP) for gene transfer. The studies investigated the effects of both direct transfection of the SON and indirect transfection (i.e., via retrograde transport) of SFO neurons. The SON of rats were injected with Ad (2 x 10(6) pfu) and sacrificed 1-7 days later for cell culture of the SON and of the SFO. In the SON, GFP fluorescence was visualized in both neuronal and nonneuronal cells while only neurons in the SFO expressed GFP. Successful in vitro transfection of cultured cells from the SON and SFO was also achieved with Ad (2 x 10(6) to 2 x 10(8) pfu). The expression of GFP in in vitro transfected cells was higher in nonneuronal (approximately 28% in SON and SFO) than neuronal (approximately 4% in SON and 10% in SFO) cells. The expression of GFP was time and viral concentration related. No apparent alterations in cellular morphology of transfected cells were detected and electrophysiological characterization of transfected cells was similar between GFP-expressing and nonexpressing neurons. We conclude that (1) GFP is an effective marker for gene transfer in living SON and SFO cells, (2) Ad infects both neuronal and nonneuronal cells, (3) Ad is taken up by axonal projections from the SON and retrogradely transported to the SFO where it is expressed at detectable levels, and (4) Ad does not adversely affect neuronal viability. These results demonstrate the feasibility of using adenoviral vectors to deliver genes to the SFO-SON axis. Copyright 1998 Academic Press.

NASA Discipline Regulatory Physiology

Adenovirus-mediated gene delivery to cells of the magnocellular hypothalamo-neurohypophyseal system

The objective of the present study was to define the optimum conditions for using replication-defective adenovirus (Ad) to transfer the gene for the green fluorescent protein (GFP) to the hypothalamic paraventricular (PVN) and supraoptic (SON) nuclei and cells of the neurohypophysis (NH). As indicated by characterizing cell survival over 15 days in culture and in electrophysiological whole cell patch-clamp studies, viral concentrations up to 2 x 10(7) pfu/coverslip did not affect viability of transfected PVN and NH cultured cells from preweanling rats. At 2 x 10(7) pfu, GFP gene expression was higher (40% of GFP-positive cells) and more sustained (up to 15 days). Using a stereotaxic approach in adult rats, we were able to directly transduce the PVN, SON, and NH and visualize gene expression in coronal brain slices and in the pituitary 4 days after injection of Ad. In animals receiving NH injections of Ad, the virus was retrogradely transported to PVN and SON neurons as indicated by the appearance of GFP-positive neurons in cultures of dissociated cells from those brain nuclei and by polymerase chain reaction and Western blot analyses of PVN and SON tissues. Adenoviral concentrations of up to 8 x 10(6) pfu injected into the NH did not affect cell viability and did not cause inflammatory responses. Adenoviral injection into the pituitary enabled the selective delivery of genes to the soma of magnocellular neurons. The experimental approaches described here provide potentially useful strategies for the treatment of disordered expression of the hormones vasopressin or oxytocin. Copyright 2000 Academic Press.

Non-NASA Center

Characterization of ionic currents of cells of the subfornical organ that project to the supraoptic nuclei

The subfornical organ (SFO) is a forebrain structure that converts peripheral blood-borne signals reflecting the hydrational state of the body to neural signals and then through efferent fibers conveys this information to several central nervous system structures. One of the forebrain areas receiving input from the SFO is the supraoptic nucleus (SON), a source of vasopressin synthesis and control of release from the posterior pituitary. Little is known of the transduction and transmission processes by which this conversion of systemic information to brain input occurs. As a step in elucidating these mechanisms, the present study characterized the ionic currents of dissociated cells of the SFO that were identified as neurons that send efferents to the SON. A retrograde tracer was injected into the SON area in eleven-day-old rats. After three days for retrograde transport of the label, the SFOs of these animals were dissociated and plated for tissue culture. The retrograde tracer was used to identify the soma of SFO cells projecting to the SON so that voltage-dependent ionic currents using whole-cell voltage clamp methods could be studied. The three types of currents in labeled SFO neurons were characterized as a 1) rapid, transient inward current that can be blocked by tetrodotoxin (TTX) characteristic of a sodium current; 2) slow-onset sustained outward current that can be blocked by tetraethylammonium (TEA) characteristic of a delayed rectifier potassium current; and 3) remaining outward current that has a rapid-onset and transient characteristic of a potassium A-type current. Copyright 1999 Elsevier Science B.V.

Non-NASA Center

Adenovirus-mediated gene delivery to hypothalamic magnocellular neurons in mice

Vasopressin is synthesized by magnocellular neurons in supraoptic (SON) and paraventricular (PVN) hypothalamic nuclei and released by their axon terminals in the neurohypophysis (NH). With its actions as an antidiuretic hormone and vasoactive agent, vasopressin plays a pivotal role in the control of body fluids and cardiovascular homeostasis. Because of its well-defined neurobiology and functional importance, the SON/PVN-NH system is ideal to establish methods for gene transfer of genetic material into specific pathways in the mouse central nervous system. In these studies, we compared the efficiency of transferring the gene lacZ, encoding for beta-galactosidase (beta-gal), versus a gene encoding for green fluorescent protein by using replication-deficient adenovirus (Ad) vectors in adult mice. Transfection with viral concentrations up to 2 x 10(7) plaque-forming units per coverslip of NH, PVN, and SON in dissociated, cultured cells caused efficient transfection without cytotoxicity. However, over an extended period of time, higher levels (50% to 75% of the cells) of beta-gal expression were detected in comparison with green fluorescent protein (5% to 50% of the cells). With the use of a stereotaxic approach, the pituitary glands of mice were injected with Ad (4 x 10(6) plaque-forming units). In material from these animals, we were able to visualize the expression of the beta-gal gene in the NH and in magnocellular neurons of both the PVN and SON. The results of these experiments indicate that Ad-Rous sarcoma virus promoter-beta-gal is taken up by nerve terminals at the injection site (NH) and retrogradely transported to the soma of the neurons projecting to the NH. We conclude that the application of these experimental approaches will provide powerful tools for physiological studies and potential approaches to deliver therapeutic genes to treat diseases.

Non-NASA Center

Statistical and Scaling Properties of Remotely-Sensed Soil Moisture in Two Contrasting Domains in the North American Monsoon Region

Characterizing soil moisture (theta) variability is important for inferring high-resolution information from coarse estimates provided by remote sensors. In this study, we analyze the spatial variability and scale invariance of high-resolution theta estimates collected in two contrasting semiarid areas, Arizona (AZ) and Sonora (SON), during the Soil Moisture Experiment - North American Monsoon in 2004 (SMEX04- NAME). Results reveal that as the mean theta condition ( ) becomes drier, the spatial standard deviation becomes smaller in both domains. The coefficient of variation of theta decreases with in SON, but does not display a clear tendency with in AZ. We also found the presence of scale invariance and multifractality in the range of support scales from 51.2 km to 0.8 km for all soil moisture fields in the two regions. The multifractal properties of theta are clearly linked to in SON, while the relation is affected by more dispersion in AZ. We argue this is due to differences in the dynamic (rainfall) and static (vegetation) controls on theta in the two domains.

Mascaro, Giuseppe

Comparison of Monthly IMERG Precipitation Estimates with PACRAIN Atoll Observations

Satellite-based precipitation estimates provide valuable information where surface observations are not readily available, especially over the large expanses of the ocean where in situ precipitation observations are very sparse. This study compares monthly precipitation estimates from the Integrated Multisatellite Retrievals for GPM (IMERG) with gauge observations from 37 low-lying atolls from the Pacific Rainfall Database for the period June 2000–August 2020. Over the analysis period, IMERG estimates are slightly higher than the atoll observations by 0.67% with a monthly correlation of 0.68. Seasonally, DJF shows excellent agreement with a near-zero bias, while MAM shows IMERG is low by 4.6%, and JJA is high by 1.2%. SON exhibits the worst performance, with IMERG overestimating by 6.5% compared to the atolls. The seasonal correlations are well contained in the range 0.67–0.72, with the exception of SON at 0.62. Furthermore, SON has the highest RMSE at 4.70 mm day^(−1), making it the worst season for all metrics. Scatterplots of IMERG versus atolls show IMERG, on average, is generally low for light precipitation accumulations and high for intense precipitation accumulations, with best agreement at intermediate rates. Seasonal variations exist at light and intermediate rate accumulations, but IMERG consistently overestimates at intense precipitation rates. The differences between IMERG and atolls vary over time but do not exhibit any discernible trend or dependence on atoll population. The PACRAIN atoll gauges are not wind-loss corrected, so application of an appropriate adjustment would increase the precipitation amounts compared to IMERG. These results provide useful insight to users as well as valuable information for future improvements to IMERG.

David T. Bolvin

Central Renin Injections: Effects on Drinking and Expression of Immediate Early Genes

This study investigated the drinking response and the expression of Fos- and Egr-1-immunoreactivity (Fos-ir, Egr-1-ir) in the brain induced by endogenous angiotensin generated by intracerebroventricular (i.c.v.) injection of renin. Renin induced Fos-ir in the subformical organ (SFO), median preoptic (MnPO), supraoptic and paraventricular nuclei (SON and PVN), area postrema (AP), nuclei of the solitary tract (NTS) and lateral parabrachial nuclei (LPBN). Renin-induced Egr-1-ir exhibited a similar pattern of distribution as that observed for Fos-ir. The dose of i.c.v. renin that induced expression of immediate early gene (IEG) product immunoreactivity also produced vigorous drinking. When renin-injected rats were pretreated with captopril, an angiotensin converting enzyme inhibitor, drinking was blocked. With the same captopril pretreatment, both Fos- and Egr-1-ir in the SFO, MnPO, SON, PVN, AP and LPBN were also significantly reduced.

Xu, Zhice

Water deprivation-induced sodium appetite: humoral and cardiovascular mediators and immediate early genes

Adult rats deprived of water for 24-30 h were allowed to rehydrate by ingesting only water for 1-2 h. Rats were then given access to both water and 1.8% NaCl. This procedure induced a sodium appetite defined by the operational criteria of a significant increase in 1.8% NaCl intake (3.8 +/- 0.8 ml/2 h; n = 6). Expression of Fos (as assessed by immunohistochemistry) was increased in the organum vasculosum of the lamina terminalis (OVLT), median preoptic nucleus (MnPO), subfornical organ (SFO), and supraoptic nucleus (SON) after water deprivation. After rehydration with water but before consumption of 1.8% NaCl, Fos expression in the SON disappeared and was partially reduced in the OVLT and MnPO. However, Fos expression did not change in the SFO. Water deprivation also 1) increased plasma renin activity (PRA), osmolality, and plasma Na+; 2) decreased blood volume; and 3) reduced total body Na+; but 4) did not alter arterial blood pressure. Rehydration with water alone caused only plasma osmolality and plasma Na+ concentration to revert to euhydrated levels. The changes in Fos expression and PRA are consistent with a proposed role for ANG II in the control of the sodium appetite produced by water deprivation followed by rehydration with only water.

Non-NASA Center

Realizing the Dream of Flight: Biographical Essays in Honor of the Centennial of Flight, 1903-2003

While growing up in Cedar Rapids, Iowa, Milton Wright, The Wright Brothers Father, liked to purchase toys for his sons that he hoped would stimulate their imagination. One of the most memorable gifts was a toy helicopter that was designed by the French aeronautical experimenter Alphonse P naud. Milton gave his sons this gift in 1878, and, though it was a simple device with a stick bound to a four-blade rotor set in a spindle, it had the intended effect it caused them to dream. Twenty-five years separated the gift of this toy and their invention of the airplane, yet the Wright brothers were convinced it had exerted an important influence. Tom Crouch argued in The Bishop's Boys that toys like these perfectly illustrated the significance of play for technological innovation. He wrote, rotary-wing toys were to intrigue and inspire generations of children, a few of whom would, as adults, attempt to realize the dream of flight for themselves. If the first powered flight on 17 December 1903 represented a childhood dream realized, it was only the first step in the rapid evolution of the airplane from their flimsy kite-like contraption of wood and cloth to jet airliners and rockets in space. And, as extraordinary as the achievement of powered flight seemed in 1903, before the end of the century, space travel also would become a dream realized. Soviet astronaut Yuri Gagarin first circumnavigated Earth in April 1961, and, eight years later, American astronauts took the first steps for humankind on the Moon. It is with great pleasure that we introduce Realizing the Dream: Biographical Essays in Honor of the Centennial of Flight. These essays in celebration of the Wright brothers first flight 100 years ago grew out of presentations by a group of prominent scholars in 2003 at a conference sponsored by the NASA History Division and held at the Great Lakes Science Center in Cleveland, Ohio. The volume focuses on the careers of some of the many men and women who helped to realize the dream of flight both through the atmosphere and beyond.

Dawson, Virginia P.

The Influence of Indian Ocean Atmospheric Circulation on Warm Pool Hydroclimate During the Holocene Epoch

Existing paleoclimate data suggest a complex evolution of hydroclimate within the Indo-Pacific Warm Pool (IPWP) during the Holocene epoch. Here we introduce a new leaf wax isotope record from Sulawesi, Indonesia and compare proxy water isotope data with ocean-atmosphere general circulation model (OAGCM) simulations to identify mechanisms influencing Holocene IPWP hydroclimate. Modeling simulations suggest that orbital forcing causes heterogenous changes in precipitation across the IPWP on a seasonal basis that may account for the differences in time-evolution of the proxy data at respective sites. Both the proxies and simulations suggest that precipitation variability during the September-November (SON) season is important for hydroclimate in Borneo. The preeminence of the SON season suggests that a seasonally lagged relationship between the Indian monsoon and Indian Ocean Walker circulation influences IPWP hydroclimatic variability during the Holocene.

simulation

Prescribing Stratospheric Chemistry Overestimates Southern Hemisphere Climate Change During Austral Spring in Response to Quadrupled CO2

The interaction of stratospheric chemistry with a changing climate from an abrupt CO2 quadrupling is assessed using the coupled atmosphere–ocean Goddard Earth Observing System Chemistry-Climate Model (GEOSCCM). Two abrupt 4 × CO2 experiments were performed, one with interactive stratospheric chemistry and the other with a prescribed stratospheric chemistry that does not simulate stratospheric ozone response to 4 × CO2. The interactive and prescribed chemistry experiments simulate similar global mean surface temperature change. Nevertheless, interactive chemistry is critical to capture the Southern Hemisphere tropospheric midlatitude jet response to 4 × CO2. When stratospheric ozone response to 4 × CO2 is neglected, GEOSCCM overestimates Southern Hemisphere tropospheric circulation change. This stratospheric chemistry-induced climate impact has large seasonal variability. During the austral spring season September–October–November (SON), prescribed chemistry yields a stronger poleward shift and intensification of the Southern Hemisphere midlatitude tropospheric jet, surface wind stress, and the Southern Ocean meridional overturning circulation than occurs with interactive chemistry. In other seasons interactive and prescribed chemistry have similar effects on the Southern Hemisphere circulation. The seasonality of stratospheric chemistry-induced climate impact is related to the seasonality of Antarctic lower stratospheric ozone response to 4 × CO2. In contrast to this stratospheric ozone response to 4 × CO2, stratospheric ozone recovery from decline of the ozone depleting substances has its largest impact on the Southern Hemisphere tropospheric circulation in austral summer (December–January–February), but no effects in SON. It is found that the different seasonality for these two stratospheric ozone layer change scenarios is related to the different seasonality of tropopause meridional temperature gradient response.

Stratospheric ozone feedback

Efficient algorithms for dilated mappings of binary trees

The problem is addressed to find a 1-1 mapping of the vertices of a binary tree onto those of a target binary tree such that the son of a node on the first binary tree is mapped onto a descendent of the image of that node in the second binary tree. There are two natural measures of the cost of this mapping, namely the dilation cost, i.e., the maximum distance in the target binary tree between the images of vertices that are adjacent in the original tree. The other measure, expansion cost, is defined as the number of extra nodes/edges to be added to the target binary tree in order to ensure a 1-1 mapping. An efficient algorithm to find a mapping of one binary tree onto another is described. It is shown that it is possible to minimize one cost of mapping at the expense of the other. This problem arises when designing pipelined arithmetic logic units (ALU) for special purpose computers. The pipeline is composed of ALU chips connected in the form of a binary tree. The operands to the pipeline can be supplied to the leaf nodes of the binary tree which then process and pass the results up to their parents. The final result is available at the root. As each new application may require a distinct nesting of operations, it is useful to be able to find a good mapping of a new binary tree over existing ALU tree. Another problem arises if every distinct required binary tree is known beforehand. Here it is useful to hardwire the pipeline in the form of a minimal supertree that contains all required binary trees.

Iqbal, M. Ashraf

Near-Infrared Spectral Geometric Albedos of Charon and Pluto: Constraints on Charon's Surface Composition

The spectral geometric albedos of Charon and Pluto are derived at near-infrared wavelengths (1.4-2.5 jAm) from measurements obtained in 1987. Comparisons of these to theoretical calculations are used to place constraints on the identity and relative abundances of surface ices on Charon. These compari- sons suggest that widespread regions of pure CH4 ice do not occur on Charon and that if CH4 is abundant on Charon then it is large grained (-5 mm) and is likely mixed at the granular level with H20 ice, and possibly C02 ice.

Roush, Ted L.

Development of Gravity Sensitive Plant Cells (Ceratodon) in Microgravity

Protonemata of the moss Ceratodon are tip-growing cells that grow up in the dark. This cell type is unique compared to cells in almost any other organism, since the growth of the plant cell itself is completely oriented by gravity. Thus, both the processes of gravity sensing and the gravity response occur in the same cell. Gravity sensing appears to rely upon amyloplasts (starch-filled plastids) that sediment. This sedimentation occurs in specific zones and plastid zonation is complex with respect to plastid morphology, distribution, and gravity. Microtubules restrict the extent of plastid sedimentation (i.e., they are load-bearing). Light also is important since apical cells have a phytochrome-based positive phototropism, light quality influences plastid zonation and sedimentation (photomorphogenesis), and red light suppresses gravitropism at higher but not lower light intensities. Many of these processes were examined in a 16 day spaceflight experiment, "SPM-A" space moss" or "SPAM)) on STS-87 that landed in December, 1997. The work described here involves the definition of a second flight experiment that builds upon the data and questions arising from STS-87. Effort was directed towards further definition of an experiment for the Shuttle (dubbed "SOS" for "Son of SPAM"). Our current target is STS 107 that is scheduled to fly in January 2001. This definition addressed two goals of the STS107 experiment. The goals of the current experiment were to determine whether the cytoskeleton plays a role in maintaining and generating an apical (non-random) plastid distribution in microgravity and to determine the development and extent of clockwise spiral tip-growth in microgravity.

Sack, Fred D.

SIM Configuration Evolution

The Space Interferometry Mission (SIM) is a space-based 10 m baseline Michelson interferometer. Planned for launch in 2005 aboard a Delta III launch vehicle, or equivalent, its primary objective is to measure the positions of stars and other celestial objects with an unprecedented accuracy of 4 micro arc seconds. With such an instrument, tremendous advancement can be expected in our understanding of stellar and galactic dynamics. Using triangulation from opposite sides of the orbit around the sun (i.e. by using parallax) one can measure the distance to any observable object in our galaxy. By directly measuring the orbital wobble of nearby stars, the mass and orbit of planets can be determined over a wide range of parameters. The distribution of velocity within nearby galaxies will be measurable. Observations of these and other objects will improve the calibration of distance estimators by more than an order of magnitude. This will permit a much better determination of the Hubble Constant as well as improving our overall understanding of the evolution of the universe. SIM has undergone several transformations, especially over the past year and a half since the start of Phase A. During this phase of a project, it is desirable to perform system-level trade studies, so the substantial evolution of the design that has occurred is quite appropriate. Part of the trade-off process has addressed two major underlying architectures: SIM Classic; and Son of SIM. The difference between these two architectures is related to the overall arrangement of the optical elements and the associated metrology system. Several different configurations have been developed for each architecture. Each configuration is the result of design choices that are influenced by many competing considerations. Some of the more important aspects will be discussed. The Space Interferometry Mission has some extremely challenging goals: millikelvin thermal stability, nanometer stabilization of optics, picometer measurement of wavefront, and others. In order to meet these goals, a significant amount of technological development is required. Although there has been a program operating for about a decade developing technologies specifically to address the challenges of space-based interferometry, there still remains a tremendous effort to achieve the incredible accuracy required of SIM. The projected viability of some of these areas has influenced design choices during the evolution of the many configurations that have been developed. For instance, the perceived complexity of the IR laser metrology system used to measure and control the positions of key optical elements was the strongest discriminator between the two architectures, and led to a decision to select SOS rather than Classic in early 1998. More recently, an appreciation of the sensitivity to beam-walk within the SOS architecture is forcing a reconsideration of that decision. At the time of submission of this abstract, there is some hope that a full-aperture metrology system may alleviate this issue. In addition to describing the current configuration of SIM, the influence of a few selected areas on the evolution of the configuration will be discussed.

Aaron, Kim M.

NASA's Lessons Learned and Technical Standards: A Logical Marriage

Lessons Learned have been the basis for our accomplishments throughout the ages. They have been passed down from father to son, mother to daughter, teacher to pupil, and older to younger worker. Lessons Learned have also been the basis for NASA's accomplishments for more than forty years. Both government and industry have long recognized the need to systematically document and utilize the knowledge gained from past experiences in order to avoid the repetition of failures and mishaps. Lessons Learned have formed the foundation for discoveries, inventions, improvements, textbooks, and Technical Standards.

Gill, Paul