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At least 217 records · Page 12
All sourcesNASA NTRSDOE OSTI
NASA NTRS2000

Bioenergetics and mitochondrial transmembrane potential during differentiation of cultured osteoblasts

To evaluate the relationship between osteoblast differentiation and bioenergetics, cultured primary osteoblasts from fetal rat calvaria were grown in medium supplemented with ascorbate to induce differentiation. Before ascorbate treatment, the rate of glucose consumption was 320 nmol. h(-1). 10(6) cells(-1), respiration was 40 nmol. h(-1). 10(6) cells(-1), and the ratio of lactate production to glucose consumption was approximately 2, indicating that glycolysis was the main energy source for immature osteoblasts. Ascorbate treatment for 14 days led to a fourfold increase in respiration, a threefold increase in ATP production, and a fivefold increase in ATP content compared with that shown in immature cells. Confocal imaging of mitochondria stained with a transmembrane potential-sensitive vital dye showed that mature cells possessed abundant amounts of high-transmembrane-potential mitochondria, which were concentrated near the culture medium-facing surface. Acute treatment of mature osteoblasts with metabolic inhibitors showed that the rate of glycolysis rose to maintain the cellular energy supply constant. Thus progressive differentiation coincided with changes in cellular metabolism and mitochondrial activity, which are likely to play key roles in osteoblast function.

NASA Center ARC↗
NASA NTRS1999

Effects of proton and gamma radiation on lymphocyte populations and acute response to antigen

BACKGROUND: The clinical use of proton radiation in the management of cancer, as well as benign disorders, is rapidly increasing. The major goal of this study was to compare the effects of proton and gamma (60Co) radiation on cell-mediated and humoral immunological parameters. MATERIALS AND METHODS: C57BL/6 mice were exposed to a single dose of 3 Gray (Gy) protons or gamma-rays and intraperitoneally injected 1 day later with sheep red blood cells (sRBC). On 4, 10, 15, and 29 days after exposure, subsets from each group were euthanised; nonirradiated controls (with and without sRBC injection) were included. Body and relative spleen weights, leukocyte counts, spontaneous blastogenesis, lymphocyte populations, and anti-sRBC titers were evaluated. RESULTS: The data showed significant depression (p < 0.05) in nearly all assays on days 4 and 10 after irradiation. B lymphocytes (CD19+) were the most radiosensitive, although reconstitution back to normal levels was observed by day 15. T cell (CD3+) and T helper cell (CD4+) recovery was evident by day 29, whereas the T cytotoxic cell (CD8+) count remained significantly below normal. Natural killer cells (NK1.1+) were relatively radioresistant. Anti-sRBC antibody production was slow and low titers were obtained after irradiation. No significant differences were noted between the two types of radiation. CONCLUSIONS: Taken together, the data show that whole-body irradiation with protons or gamma-rays, at the dose employed, results in marked, but transient, immunosuppression. However, at the time points of testing and with the assays used, little or no differences were found between the two forms of radiation.

NASA Discipline Radiation Health↗
NASA NTRS2003

Phytochromes play a role in phototropism and gravitropism in Arabidopsis roots

Phototropism as well as gravitropism plays a role in the oriented growth of roots in flowering plants. In blue or white light, roots exhibit negative phototropism, but red light induces positive phototropism in Arabidopsis roots. Phytochrome A (phyA) and phyB mediate the positive red-light-based photoresponse in roots since single mutants (and the double phyAB mutant) were severely impaired in this response. In blue-light-based negative phototropism, phyA and phyAB (but not phyB) were inhibited in the response relative to the WT. In root gravitropism, phyB and phyAB (but not phyA) were inhibited in the response compared to the WT. The differences observed in tropistic responses were not due to growth limitations since the growth rates among all the mutants tested were not significantly different from that of the WT. Thus, our study shows that the blue-light and red-light systems interact in roots and that phytochrome plays a key role in plant development by integrating multiple environmental stimuli. c2003 COSPAR. Published by Elsevier Ltd. All rights reserved.

Non-NASA Center↗
NASA NTRS1990

Photomorphogenesis and photoassimilation in soybean and sorghum grown under broad spectrum or blue-deficient light sources

The role of blue light in plant growth and development was investigated in soybean (Glycine max [L.] Merr. cv Williams) and sorghum (Sorghum bicolor [L.] Moench. cv Rio) grown under equal photosynthetic photon fluxes (approximately 500 micromoles per square meter per second) from broad spectrum daylight fluorescent or blue-deficient, narrow-band (589 nanometers) low pressure sodium (LPS) lamps. Between 14 and 18 days after sowing, it was possible to relate adaptations in photosynthesis and leaf growth to dry matter accumulation. Soybean development under LPS light was similar in several respects to that of shaded plants, consistent with an important role for blue light photoreceptors in regulation of growth response to irradiance. Thus, soybeans from LPS conditions partitioned relatively more growth to leaves and maintained higher average leaf area ratios (mean LAR) that compensated lower net assimilation rates (mean NAR). Relative growth rates were therefore comparable to plants from daylight fluorescent lamps. Reductions in mean NAR were matched by lower rates of net photosynthesis (A) on an area basis in the major photosynthetic source (first trifoliolate) leaf. Lower A in soybean resulted from reduced leaf dry matter per unit leaf area, but lower A under LPS conditions in sorghum correlated with leaf chlorosis and reduced total nitrogen (not observed in soybean). In spite of a lower A, mean NAR was larger in sorghum from LPS conditions, resulting in significantly greater relative growth rates (mean LAR was approximately equal for both light conditions). Leaf starch accumulation rate was higher for both species and starch content at the end of the dark period was elevated two- and three-fold for sorghum and soybean, respectively, under LPS conditions. Possible relations between starch accumulation, leaf export, and plant growth in response to spectral quality were considered.

NASA Discipline Life Support Systems↗
NASA NTRS2000

Endogenous circulating sympatholytic factor in orthostatic intolerance

Sympathotonic orthostatic hypotension (SOH) is an idiopathic syndrome characterized by tachycardia, hypotension, elevated plasma norepinephrine, and symptoms of orthostatic intolerance provoked by assumption of an upright posture. We studied a woman with severe progressive SOH with blood pressure unresponsive to the pressor effects of alpha(1)-adrenergic receptor (AR) agonists. We tested the hypothesis that a circulating factor in this patient interferes with vascular adrenergic neurotransmission. Preincubation of porcine pulmonary artery vessel rings with patient plasma produced a dose-dependent inhibition of vasoconstriction to phenylephrine in vitro, abolished vasoconstriction to direct electrical stimulation, and had no effect on nonadrenergic vasoconstrictive stimuli (endothelin-1), PGF-2alpha (or KCl). Preincubation of vessels with control plasma was devoid of these effects. SOH plasma inhibited the binding of an alpha(1)-selective antagonist radioligand ([(125)I]HEAT) to membrane fractions derived from porcine pulmonary artery vessel rings, rat liver, and cell lines selectively overexpressing human ARs of the alpha(1B) subtype but not other AR subtypes (alpha(1A) and alpha(1D)). We conclude that a factor in SOH plasma can selectively and irreversibly inhibit adrenergic ligand binding to alpha(1B) ARs. We propose that this factor contributes to a novel pathogenesis for SOH in this patient. This patient's syndrome represents a new disease entity, and her plasma may provide a unique tool for probing the selective functions of alpha(1)-ARs.

Case Reports↗
NASA NTRS2005

Strawman Philosophical Guide for Developing International Network of GPM GV Sites

The creation of an international network of ground validation (GV) sites that will support the Global Precipitation Measurement (GPM) Mission's international science programme will require detailed planning of mechanisms for exchanging technical information, GV data products, and scientific results. An important component of the planning will be the philosophical guide under which the network will grow and emerge as a successful element of the GPM Mission. This philosophical guide should be able to serve the mission in developing scientific pathways for ground validation research which will ensure the highest possible quality measurement record of global precipitation products. The philosophical issues, in this regard, partly stem from the financial architecture under which the GV network will be developed, i.e., each participating country will provide its own financial support through committed institutions -- regardless of whether a national or international space agency is involved.At the 1st International GPM Ground Validation Workshop held in Abingdon, UK in November-2003, most of the basic tenants behind the development of the international GV network were identified and discussed. Therefore, with this progress in mind, this presentation is intended to put forth a strawman philosophical guide supporting the development of the international network of GPM GV sites, noting that the initial progress has been reported in the Proceedings of the 1st International GPM GV Workshop -- available online. The central philosophical issues themselves, all flow from the fact that each participating institution can only bring to the table, GV facilities and scientific personnel that are affordable to the sanctioning (funding) national agency (be that a research, research-support, or operational agency). This situation imposes on the network, heterogeneity in the measuring sensors, data collection periods, data collection procedures, data latencies, and data reporting capabilities. Therefore, in order for the network to be effective in supporting the central scientific goals of the GPM mission, there must be a basic agreed upon doctrine under which the network participants function vis-a-vis: (1) an overriding set of general scientific requirements, (2) a minimal set of policies governing the free flow of GV data between the scientific participants, (3) a few basic definitions concerning the prioritization of measurements and their respective value to the mission, (4) a few basic procedures concerning data formats, data reporting procedures, data access, and data archiving, and (5) a simple means to differentiate GV sites according to their level of effort and ability to perform near real-time data acquisition - data reporting tasks. Most important, in case they choose to operate as a near real-time data collection-data distribution site, they would be expected to operate under a fairly narrowly defined protocol needed to ensure smooth GV support operations. This presentation will suggest measures responsive to items (1) - (5) from which to proceed,. In addition, this presentation will seek to stimulate discussion and debate concerning how much heterogeneity is tolerable within the eventual GV site network, given that the any individual GV site can only be considered scientifically useful if it supports the achievement of the central GPM Mission goals. Only ground validation research that has a direct connection to the space mission should be considered justifiable given the overarching scientific goals of the mission. Therefore each site will have to seek some level of accommodation to what the GPM Mission requires in the way of retrieval error characterization, retrieval error detection and reporting, and generation of GV data products that support assessment and improvement of the mission's standard precipitation retrieval algorithms. These are all important scientific issues that will be best resolved in open scientific debate.

Smith, Eric A.↗
NASA NTRS1996

Complete disintegration of the microtubular cytoskeleton precedes its auxin-mediated reconstruction in postmitotic maize root cells

The inhibitory action of 0.1 microM auxin (IAA) on maize root growth was closely associated with a rapid and complete disintegration of the microtubular (MT) cytoskeleton, as visualized by indirect immunofluorescence of tubulin, throughout the growth region. After 30 min of this treatment, only fluorescent spots were present in root cells, accumulating either around nuclei or along cell walls. Six h later, in addition to some background fluorescence, dense but partially oriented oblique or longitudinal arrays of cortical MTs (CMTs) were found in most growing cells of the root apex. After 24 h of treatment, maize roots had adapted to the auxin, as inferred from the slowly recovering elongation rate and from the reassembly of a dense and well-ordered MT cytoskeleton which showed only slight deviations from that of the control root cells. Taxol pretreatment (100 microM, 24 h) prevented not only the rapid auxin-mediated disintegration of the MT cytoskeleton but also a reorientation of the CMT arrays, from transversal to longitudinal. The only tissue to show MTs in their cells throughout the auxin treatment was the epidermis. Significant resistance of transverse CMT arrays in these cells towards auxin was confirmed using a higher auxin concentration (100 microM, 24 h). The latter auxin dose also revealed inter-tissue-specific responses to auxin: outer cortical cell files reoriented their CMTs from the transversal to longitudinal orientation, whereas inner cortical cell files lost their MTs. This high auxin-mediated response, associated with the swelling of root apices, was abolished with the pretreatment of maize root with taxol.

Zea mays/cytology/drug effects/growth & developmen↗
NASA NTRS1986

Do the higher oxidation states of the photosynthetic O2-evolving system contain bound H2O?

A modified mass spectrometer was used to determine whether the higher oxidation states of the photosynthetic O2-evolving system contain substrate water that is not freely exchangeable with the external medium. Our data indicated that the higher oxidation states contain no appreciable bound, non-exchangeable H2O. This suggests that H2O oxidation takes place via a rapid, concerted, all-or-none mechanism rather than by a mechanism involving stable, partially oxidized, H2O-derived intermediates. These findings set definite constraints on possible mechanisms of O2 evolution.

NASA Discipline Life Support Systems↗
NASA NTRS1992

Effects of dimethyl prostaglandin A1 on herpes simplex virus and human immunodeficiency virus replication

We have investigated the direct effect of dimethyl prostaglandin A1 (dmPGA1) on the replication of herpes simplex virus (HSV) and human immunodeficiency virus type 1 (HIV-1). dmPGA1 significantly inhibited viral replication in both HSV and HIV infection systems at concentrations of dmPGA1 that did not adversely alter cellular DNA synthesis. The 50% inhibitory concentration (ID50) for several HSV type 1 (HSV-1) strains ranged from 3.8 to 5.6 micrograms/ml for Vero cells and from 4.6 to 7.3 micrograms/ml for human foreskin fibroblasts. The ID50s for two HSV-2 strains varied from 3.8 to 4.5 micrograms/ml for Vero cells; the ID50 was 5.7 micrograms/ml for human foreskin fibroblasts. We found that closely related prostaglandins did not have the same effect on the replication of HSV; dmPGE2 and dmPGA2 caused up to a 60% increase in HSV replication compared with that in untreated virus-infected cells. HIV-1 replication in acutely infected T cells (VB line) and chronically infected macrophages was assessed by quantitative decreases in p24 concentration. The effective ID50s were 2.5 micrograms/ml for VB cells acutely infected with HIV-1 and 5.2 micrograms/m for chronically infected macrophages. dmPGA1 has an unusual broad-spectrum antiviral activity against both HSV and HIV-1 in vitro and offers a new class of potential therapeutic agents for in vivo use.

Non-NASA Center↗
NASA NTRS2001

TGF-beta1 release from biodegradable polymer microparticles: its effects on marrow stromal osteoblast function

BACKGROUND: Controlled release of transforming growth factor-beta1 (TGF-beta1) to a bone defect may be beneficial for the induction of a bone regeneration cascade. The objectives of this work were to assess the feasibility of using biodegradable polymer microparticles as carriers for controlled TGF-beta1 delivery and the effects of released TGF-beta1 on the proliferation and differentiation of marrow stromal cells in vitro. METHODS: Recombinant human TGF-beta1 was incorporated into microparticles of blends of poly(DL-lactic-co-glycolic acid) (PLGA) and poly(ethylene glycol) (PEG). Fluorescein isothiocynate-labeled bovine serum albumin (FITC-BSA) was co-encapsulated as a porogen. The effects of PEG content (0, 1, or 5% by weight [wt%]) and buffer pH (3, 5, or 7.4) on the protein release kinetics and the degradation of PLGA were determined in vitro for as long as 28 days. Rat marrow stromal cells were seeded on a biodegradable poly(propylene fumarate) (PPF) substrate. The dose response and biological activity of released TGF-beta1 was determined after 3 days in culture. The effects of TGF-beta1 released from PLGA/PEG microparticles on marrow stromal cell proliferation and osteoblastic differentiation were assessed during a 21-day period. RESULTS: TGF-beta1 was encapsulated along with FITC-BSA into PLGA/PEG blend microparticles and released in a multiphasic fashion including an initial burst for as long as 28 days in vitro. Increasing the initial PEG content resulted in a decreased cumulative mass of released proteins. Aggregation of FITC-BSA occurred at lower buffer pH, which led to decreased release rates of both proteins. The degradation of PLGA was increased at higher PEG content and significantly accelerated at acidic pH conditions. Rat marrow stromal cells cultured on PPF substrates showed a dose response to TGF-beta1 released from the microparticles similar to that of added TGF-beta1, indicating that the activity of TGF-beta1 was retained during microparticle fabrication and after growth factor release. At an optimal TGF-beta1 dosage of 1.0 ng/ml after 3 days, the released TGF-beta1 enhanced the proliferation and osteoblastic differentiation of marrow stromal cells over 21 days of culture, with increased total cell number, alkaline phosphatase activity, and osteocalcin production. CONCLUSIONS: PLGA/PEG blend microparticles can serve as delivery vehicles for controlled release of TGF-beta1, and the released growth factor enhances marrow stromal cell proliferation and osteoblastic differentiation in vitro. CLINICAL RELEVANCE: Controlled release of TGF-beta1 from PLGA/PEG microparticles is representative of emerging tissue engineering technologies that may modulate cellular responses to encourage bone regeneration at a skeletal defect site.

Non-NASA Center↗
NASA NTRS2000

Dose-response relationship for light intensity and ocular and electroencephalographic correlates of human alertness

Light can elicit both circadian and acute physiological responses in humans. In a dose response protocol men and women were exposed to illuminances ranging from 3 to 9100 lux for 6.5 h during the early biological night after they had been exposed to <3 lux for several hours. Light exerted an acute alerting response as assessed by a reduction in the incidence of slow-eye movements, a reduction of EEG activity in the theta-alpha frequencies (power density in the 5-9 Hz range) as well as a reduction in self-reported sleepiness. This alerting response was positively correlated with the degree of melatonin suppression by light. In accordance with the dose response function for circadian resetting and melatonin suppression, the responses of all three indices of alertness to variations in illuminance were consistent with a logistic dose response curve. Half of the maximum alerting response to bright light of 9100 lux was obtained with room light of approximately 100 lux. This sensitivity to light indicates that variations in illuminance within the range of typical, ambient, room light (90-180 lux) can have a significant impact on subjective alertness and its electrophysiologic concomitants in humans during the early biological night.

Clinical Trial↗
NASA NTRS1993

A wall interference assessment/correction system

A Wall Signature method originally developed by Hackett was selected to be adapted for the Ames 12-ft Wind Tunnel Wall Interference Assessment/Correction (WIAC) System in the project. This method uses limited measurements of the static pressure at the wall, in conjunction with the solid wall boundary condition, to determine the strength and distribution of singularities representing the test article. The singularities are used in turn for estimating wall interference at the model location. The lifting interference will be treated separately by representing in a horseshoe vortex system for the model's lifting effects. The development and implementation of a working prototype will be completed, delivered, and documented with a software manual. The WIAC code will be validated by conducting numerically simulated experiments rather than actual wind tunnel experiments. The simulations will be used to generate both free-air and confined wind-tunnel flow fields for each of the test articles over a range of test configurations. Specifically, the pressure signature at the test section wall will be computed for the tunnel case to provide the simulated 'measured' data. These data will serve as the input for the WIAC method - Wall Signature method. The performance of the WIAC method then may be evaluated by comparing the corrected parameters with those for the free-air simulation. The following two additional tasks are included in the supplement No. 1 to the basic Grant. On-line wall interference calculation: The developed wall signature method (modified Hackett's method) for Ames 12-ft Tunnel will be the pre-computed coefficients which facilitate the on-line calculation of wall interference; and support system effects estimation: The effects on the wall pressure measurements due to the presence of the model support systems will be evaluated.

Lo, Ching F.↗
NASA NTRS1993

Accuracy of quantum sensors measuring yield photon flux and photosynthetic photon flux

Photosynthesis is fundamentally driven by photon flux rather than energy flux, but not all absorbed photons yield equal amounts of photosynthesis. Thus, two measures of photosynthetically active radiation have emerged: photosynthetic photon flux (PPF), which values all photons from 400 to 700 nm equally, and yield photon flux (YPF), which weights photons in the range from 360 to 760 nm according to plant photosynthetic response. We selected seven common radiation sources and measured YPF and PPF from each source with a spectroradiometer. We then compared these measurements with measurements from three quantum sensors designed to measure YPF, and from six quantum sensors designed to measure PPF. There were few differences among sensors within a group (usually <5%), but YPF values from sensors were consistently lower (3% to 20%) than YPF values calculated from spectroradiometric measurements. Quantum sensor measurements of PPF also were consistently lower than PPF values calculated from spectroradiometric measurements, but the differences were <7% for all sources, except red-light-emitting diodes. The sensors were most accurate for broad-band sources and least accurate for narrow-band sources. According to spectroradiometric measurements, YPF sensors were significantly less accurate (>9% difference) than PPF sensors under metal halide, high-pressure sodium, and low-pressure sodium lamps. Both sensor types were inaccurate (>18% error) under red-light-emitting diodes. Because both YPF and PPF sensors are imperfect integrators, and because spectroradiometers can measure photosynthetically active radiation much more accurately, researchers should consider developing calibration factors from spectroradiometric data for some specific radiation sources to improve the accuracy of integrating sensors.

NASA Center KSC↗
NASA NTRS1992

Ionizing radiation exposure of LDEF (pre-recovery estimates)

The long duration exposure facility (LDEF), launched into a 258 nautical mile orbit with an inclination of 28.5 degrees, remained in space for nearly 6 yr. The 21,500 lb NASA satellite was one of the largest payloads ever deployed by the Space Shuttle. LDEF completed 32,422 orbits and carried 57 major experiments representing more than 200 investigators from 33 private companies, 21 universities and nine countries. The experiments covered a wide range of disciplines including basic science, electronics, optics, materials, structures and power and propulsion. A number of the experiments were specifically designed to measure the radiation environment. These experiments are of specific interest, since the LDEF orbit is essentially the same as that of the Space Station Freedom. Consequently, the radiation measurements on LDEF will play a significant role in the design of radiation shielding of the space station. The contributions of the various authors presented here attempt to predict the major aspects of the radiation exposure received by the various LDEF experiments and therefore should be helpful to investigators who are in the process of analyzing experiments which may have been affected by exposure to ionizing radiation. The paper discusses the various types and sources of ionizing radiation including cosmic rays, trapped particles (both protons and electrons) and secondary particles (including neutrons, spallation products and high-LET recoils), as well as doses and LET spectra as a function of shielding. Projections of the induced radioactivity of LDEF are also discussed.

Flight Experiment↗
NASA NTRS1995

Parallel automated adaptive procedures for unstructured meshes

Consideration is given to the techniques required to support adaptive analysis of automatically generated unstructured meshes on distributed memory MIMD parallel computers. The key areas of new development are focused on the support of effective parallel computations when the structure of the numerical discretization, the mesh, is evolving, and in fact constructed, during the computation. All the procedures presented operate in parallel on already distributed mesh information. Starting from a mesh definition in terms of a topological hierarchy, techniques to support the distribution, redistribution and communication among the mesh entities over the processors is given, and algorithms to dynamically balance processor workload based on the migration of mesh entities are given. A procedure to automatically generate meshes in parallel, starting from CAD geometric models, is given. Parallel procedures to enrich the mesh through local mesh modifications are also given. Finally, the combination of these techniques to produce a parallel automated finite element analysis procedure for rotorcraft aerodynamics calculations is discussed and demonstrated.

Shephard, M. S.↗
NASA NTRS1950

An Analysis of Base Pressure at Supersonic Velocities and Comparison with Experiment

In the first part of the investigation an analysis is made of the base pressure in an inviscid fluid, both for two-dimensional and axially-symmetric flow. It is shown that for two-dimensional flow, and also for the flow over a body of revolution with a cylindrical sting attached to the base, there are an infinite number of possible solutions satisfying all necessary boundary conditions at any given free-stream Mach number. For the particular case of a body having no sting attached only one solution is possible in an inviscid flow, but it corresponds to zero base drag. Accordingly, it is concluded that a strictly inviscid-fluid theory cannot be satisfactory for practical applications. Since the exact inviscid-fluid does not adequately describe the conditions of a real fluid flow, an approximate semi-empirical theory for base pressure in a viscous fluid is developed in a second part of the investigation. The semi-empirical theory is based partly on inviscid-flow calculations, and is restricted to airfoils and bodies without boattailing. In this theory an attempt is made to allow for the effects of Mach number, Reynolds number, profile shape, and type of boundary-layer flow. The results of some recent experimental measurements of base pressure in two-dimensional and axially-symmetric flow are presented for purposes of comparison. Some experimental results also are presented concerning the support interference effect of a cylindrical sting, and the interference effect of a reflected bow wave on measurements of base pressure in a supersonic wind tunnel.

Chapman, Dean R↗
NASA NTRS1997

Latent transforming growth factor beta1 activation in situ: quantitative and functional evidence after low-dose gamma-irradiation

The biological activity of transforming growth factor beta1 (TGF-beta) is controlled by its secretion as a latent complex in which it is noncovalently associated with latency-associated peptide (LAP). Activation is the extracellular process in which TGF-beta is released from LAP, and is considered to be a primary regulatory control. We recently reported rapid and persistent changes in TGF-beta immunoreactivity in conjunction with extracellular matrix remodeling in gamma-irradiated mouse mammary gland. Our hypothesis is that these specific changes in immunoreactivity are indicative of latent TGF-beta activation. In the present study, we determined the radiation dose response and tested whether a functional relationship exists between radiation-induced TGF-beta and collagen type III remodeling. After radiation exposures as low as 0.1 Gy, we detected increased TGF-beta immunoreactivity in the mammary epithelium concomitant with decreased LAP immunostaining, which are events consistent with activation. Quantitative image analysis demonstrated a significant (P=0.0005) response at 0.1 Gy without an apparent threshold and a linear dose response to 5 Gy. However, in the adipose stroma, loss of LAP demonstrated a qualitative threshold at 0.5 Gy. Loss of LAP paralleled induction of collagen III immunoreactivity in this tissue compartment. We tested whether TGF-beta mediates collagen III expression by treating animals with TGF-beta panspecific monoclonal antibody, 1D11.16, administered i.p. shortly before irradiation. Radiation-induced collagen III staining in the adipose stroma was blocked in an antibody dose-dependent manner, which persisted through 7 days postirradiation. RNase protection assay revealed that radiation-induced elevation of total gland collagen III mRNA was also blocked by neutralizing antibody treatment. These data provide functional confirmation of the hypothesis that radiation exposure leads to latent TGF-beta activation, support our interpretation of the reciprocal shift in immunoreactivity as evidence of activation, and implicate TGF-beta as a mediator of tissue response to ionizing radiation. The sensitivity of activation to low radiation doses points to a potential role for TGF-beta in orchestrating tissue response to oxidative stress. As such, radiation may be useful as a probe to delineate the consequences of latent TGF-beta activation in situ.

NASA Discipline Radiation Health↗
NASA NTRS2002

Interactions involving the Rad51 paralogs Rad51C and XRCC3 in human cells

Homologous recombinational repair of DNA double-strand breaks and crosslinks in human cells is likely to require Rad51 and the five Rad51 paralogs (XRCC2, XRCC3, Rad51B/Rad51L1, Rad51C/Rad51L2 and Rad51D/Rad51L3), as has been shown in chicken and rodent cells. Previously, we reported on the interactions among these proteins using baculovirus and two- and three-hybrid yeast systems. To test for interactions involving XRCC3 and Rad51C, stable human cell lines have been isolated that express (His)6-tagged versions of XRCC3 or Rad51C. Ni2+-binding experiments demonstrate that XRCC3 and Rad51C interact in human cells. In addition, we find that Rad51C, but not XRCC3, interacts directly or indirectly with Rad51B, Rad51D and XRCC2. These results argue that there are at least two complexes of Rad51 paralogs in human cells (Rad51C-XRCC3 and Rad51B-Rad51C-Rad51D-XRCC2), both containing Rad51C. Moreover, Rad51 is not found in these complexes. X-ray treatment did not alter either the level of any Rad51 paralog or the observed interactions between paralogs. However, the endogenous level of Rad51C is moderately elevated in the XRCC3-overexpressing cell line, suggesting that dimerization between these proteins might help stabilize Rad51C.

Non-NASA Center↗