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At least 181 records · Page 10

Effects of Dietary Iron and Gamma Radiation on the Rat Retina

A health risk of concern for NASA relates to radiation exposure and its synergistic effects with other space environmental factors, includi ng nutritional status of the crew. Astronauts consume almost three times the recommended daily allowance of iron due to the use of fortifie d foods aboard the International Space Station, with iron intake occa sionally exceeding six times the recommended values. Recently, NASA has become concerned with visual changes associated with spaceflight, a nd research is being conducted to elucidate the etiology of eye structure alterations in the spaceflight environment. Terrestrially, iron o verload is also associated with certain optic neuropathies. In additi on, due to its role in Fenton reactions, iron can potentiate oxidative stress, which is a recognized cause of cataract formation. As part o f a study investigating the combined effects of radiation exposure an d iron overload on multiple physiological systems, we focused on defining the effects of both treatments on eye biology. In this study, 12- week-old Sprague-Dawley rats were assigned to one of four experimental groups: normal iron/no radiation (Control/Sham), high iron/no radiat ion (Fe/Sham), normal iron/gamma radiation (3 Gy cumulative dose, fra ctionated at 0.375 Gy/d every other day for 16 d) (Control/Rad), and high iron/gamma radiation (Fe/Rad). Oxidative stress-induced DNA damag e, measured as concentration of the marker 8-hydroxy-2'-deoxyguanosine (8OHdG) in eye retinal tissue by enzyme-immunoanalysis did not show significant changes among treatments. However, there was an overall i ncrease in 8OHdG immunostaining density in retina sections due to radiation exposure (P = 0.05). Increased dietary iron and radiation expos ure had an interactive effect (P = 0.02) on 8OHdG immunostaining of t he retinal ganglion cell layer with iron diet increasing the signal in the group not exposed to radiation (P = 0.05). qPCR gene expression profiling of relevant target genes indicated upregulation of ferritin light chain (P = 0.09) as a result of dietary iron but no change in e xpression of the gene for ferritin heavy chain. Immunolocalization of light chain and heavy chain of the iron storage protein ferritin showed the expected distribution in the choroid, photoreceptor layer, inn er nuclear layer and in the inner plexiform layer that corresponded t o the synaptic terminals of bipolar cells. Evidence of stress and damage in the retina was also suggested by a decrease in expression of th e survival marker Bcl2 (P = 0.01) and the protective proteins clusterin (P = 0.04) and heat shock factor 1 (Hsf1, P < 0.001), as a result o f increased dietary iron. The effect of increased iron on expression of the antioxidant enzyme heme oxygenase 1 (Hmox1) had a significant interaction with the effect of radiation (P < 0.001). In summary, the results of this study indicate that both gamma radiation exposure and a moderate increase in dietary iron can contribute to deleterious cha nges in retinal health and physiology.

Morgan, Jennifer↗

Mechanisms of cell signaling by nitric oxide and peroxynitrite: from mitochondria to MAP kinases

Many of the biological and pathological effects of nitric oxide (NO) are mediated through cell signaling pathways that are initiated by NO reacting with metalloproteins. More recently, it has been recognized that the reaction of NO with free radicals such as superoxide and the lipid peroxyl radical also has the potential to modulate redox signaling. Although it is clear that NO can exert both cytotoxic and cytoprotective actions, the focus of this overview are those reactions that could lead to protection of the cell against oxidative stress in the vasculature. This will include the induction of antioxidant defenses such as glutathione, activation of mitogen-activated protein kinases in response to blood flow, and modulation of mitochondrial function and its impact on apoptosis. Models are presented that show the increased synthesis of glutathione in response to shear stress and inhibition of cytochrome c release from mitochondria. It appears that in the vasculature NO-dependent signaling pathways are of three types: (i) those involving NO itself, leading to modulation of mitochondrial respiration and soluble guanylate cyclase; (ii) those that involve S-nitrosation, including inhibition of caspases; and (iii) autocrine signaling that involves the intracellular formation of peroxynitrite and the activation of the mitogen-activated protein kinases. Taken together, NO plays a major role in the modulation of redox cell signaling through a number of distinct pathways in a cellular setting.

Non-NASA Center↗

Thermoelastic analysis of solar cell arrays and their material properties

A thermoelastic stress analysis procedure is reported for predicting the thermally induced stresses and failures in silicon solar cell arrays. A prerequisite for the analysis is the characterization of the temperature-dependent thermal and mechanical properties of the solar cell materials. Extensive material property testing was carried out in the temperature range -200 to +200 C for the filter glass, P- and N-type silicon, interconnector metals, solder, and several candidate silicone rubber adhesives. The analysis procedure is applied to several solar cell array design configurations. Results of the analysis indicate the optimum design configuration, with respect to compatible materials, effect of the solder coating, and effect of the interconnector geometry. Good agreement was found between results of the analysis and the test program.

Salama, M. A.↗

Biosentinel: Improving Desiccation Tolerance of Yeast Biosensors for Deep-Space Missions

BioSentinel is one of 13 secondary payloads to be deployed on Exploration Mission 1 (EM-1) in 2019. We will use the budding yeast Saccharomyces cerevisiae as a biosensor to determine how deep-space radiation affects living organisms and to potentially quantify radiation levels through radiation damage analysis. Radiation can damage DNA through double strand breaks (DSBs), which can normally be repaired by homologous recombination. Two yeast strains will be air-dried and stored in microfluidic cards within the payload: a wild-type control strain and a radiation sensitive rad51 mutant that is deficient in DSB repairs. Throughout the mission, the microfluidic cards will be rehydrated with growth medium and an indicator dye. Growth rates of each strain will be measured through LED detection of the reduction of the indicator dye, which correlates with DNA repair and the amount of radiation damage accumulated. Results from BioSentinel will be compared to analog experiments on the ISS and on Earth. It is well known that desiccation can damage yeast cells and decrease viability over time. We performed a screen for desiccation-tolerant rad51 strains. We selected 20 re-isolates of rad51 and ran a weekly screen for desiccation-tolerant mutants for five weeks. Our data shows that viability decreases over time, confirming previous research findings. Isolates L2, L5 and L14 indicate desiccation tolerance and are candidates for whole-genome sequencing. More time is needed to determine whether a specific strain is truly desiccation tolerant. Furthermore, we conducted an intracellular trehalose assay to test how intracellular trehalose concentrations affect or protect the mutant strains against desiccation stress. S. cerevisiae cell and reagent concentrations from a previously established intracellular trehalose protocol did not yield significant absorbance measurements, so we tested varying cell and reagent concentrations and determined proper concentrations for successful protocol use.

BioSentinel↗

Biosentinel: Improving Desiccation Tolerance of Yeast Biosensors for Deep-Space Missions

BioSentinel is one of 13 secondary payloads to be deployed on Exploration Mission 1 (EM-1) in 2019. We will use the budding yeast Saccharomyces cerevisiae as a biosensor to determine how deep-space radiation affects living organisms and to potentially quantify radiation levels through radiation damage analysis. Radiation can damage DNA through double strand breaks (DSBs), which can normally be repaired by homologous recombination. Two yeast strains will be air-dried and stored in microfluidic cards within the payload: a wild-type control strain and a radiation sensitive rad51 mutant that is deficient in DSB repairs. Throughout the mission, the microfluidic cards will be rehydrated with growth medium and an indicator dye. Growth rates of each strain will be measured through LED detection of the reduction of the indicator dye, which correlates with DNA repair and the amount of radiation damage accumulated. Results from BioSentinel will be compared to analog experiments on the ISS and on Earth. It is well known that desiccation can damage yeast cells and decrease viability over time. We performed a screen for desiccation-tolerant rad51 strains. We selected 20 re-isolates of rad51 and ran a weekly screen for desiccation-tolerant mutants for five weeks. Our data shows that viability decreases over time, confirming previous research findings. Isolates L2, L5 and L14 indicate desiccation tolerance and are candidates for whole-genome sequencing. More time is needed to determine whether a specific strain is truly desiccation tolerant. Furthermore, we conducted an intracellular trehalose assay to test how intracellular trehalose concentrations affect or protect the mutant strains against desiccation stress. S. cerevisiae cell and reagent concentrations from a previously established intracellular trehalose protocol did not yield significant absorbance measurements, so we tested varying cell and reagent concentrations and determined proper concentrations for successful protocol use.

BioSentinel↗

Lithium-doped solar cells for space use

Lithium doped p-n solar cells meet the basic requirements for space use. The efficiencies of the Lopex lithium cells which range from 10.5 to 12.8% are equal to or better than C.G. lithium cell and 10 ohms-cm n-p cell efficiencies. Reduction in the stresses introduced during boron diffusion has eliminated cell size restrictions and both 2 x 2 and 2 x 6 cm lithium doped cells have been fabricated. The Ti-Ag contacts which are tested 100% with a tape peel test, and pull tested and humidity tested on a sample basis are comparable to the Ti-Ag contacts on n-p cells.

Payne, P. A.↗

Hot-spot durability testing of amorphous cells and modules

This paper discusses the results of a study to determine the hot-spot susceptibility of amorphous-silicon (a-Si) cells and modules, and to provide guidelines for reducing that susceptibility. Amorphous-Si cells are shown to have hot-spot susceptibility levels similar to crystalline-silicon (C-Si) cells. This premise leads to the fact that the same general guidelines must apply to protecting a-Si cells from hot-spot stressing that apply to C-Si cells. Recommendations are made on ways of reducing a-Si module hot-spot susceptibility including the traditional method of using bypass diodes and a new method unique to thin-film cells, limiting the string current by limiting cell area.

Gonzalez, Charles↗

Human Adaptation Genetic Response Suites: Toward New Interventions and Countermeasures for Spaceflight

Genetic response suites in human lymphocytes in response to microgravity are important to identify and further study in order to augment human physiological adaptation to novel environments. Emerging technologies, such as DNA micro array profiling, have the potential to identify novel genes that are involved in mediating adaptation to these environments. These genes may prove to be therapeutically valuable as new targets for countermeasures, or as predictive biomarkers of response to these new environments. Human lymphocytes cultured in lg and microgravity analog culture were analyzed for their differential gene expression response. Different groups of genes related to the immune response, cardiovascular system and stress response were then analyzed. Analysis of cells from multiple donors reveals a small shared set that are likely to be essential to adaptation. These three groups focus on human adaptation to new environments. The shared set contains genes related to T cell activation, immune response and stress response to analog microgravity.

Sundaresan, A.↗

Radioadaptive Cytoprotective Pathways in the Mouse Retina

Exposure to cosmic radiation implies a risk of tissue degeneration. Radiation retinopathy is a complication of radiotherapy and exhibits common features with other retinopathies and neuropathies. Exposure to a low radiation dose elicits protective cellular events (radioadaptive response), reducing the stress of a subsequent higher dose. To assess the risk of radiation-induced retinal changes and the extent to which a small priming dose reduces this risk, we used a mouse model exposed to a source of Cs-137-gamma radiation. Gene expression profiling of retinas from non-irradiated control C57BL/6J mice (C) were compared to retinas from mice treated with a low 50 mGy dose (LD), a high 6 Gy dose (HD), and a combined treatment of 50 mGy (priming) and 6 Gy (challenge) doses (LHD). Whole retina RNA was isolated and expression analysis for selected genes performed by RTqPCR. Relevant target genes associated with cell death/survival, oxidative stress, cellular stress response and inflammation pathways, were analyzed. Cellular stress response genes were upregulated at 4 hr after the challenge dose in LHD retinas (Sirt1: 1.5 fold, Hsf1: 1.7 fold, Hspa1a: 2.5 fold; Hif1a: 1.8 fold, Bag1: 1.7). A similar trend was observed in LD animals. Most antioxidant enzymes (Hmox1, Sod2, Prdx1, Cygb, Cat1) and inflammatory mediators (NF B, Ptgs2 and Tgfb1) were upregulated in LHD and LD retinas. Expression of the pro-survival gene Bcl2 was upregulated in LD (6-fold) and LHD (4-fold) retinas. In conclusion, cytoprotective gene networks activation in the retina suggests a radioadaptive response to a priming irradiation dose, with mitigation of the deleterious effects of a subsequent high dose exposure. The enhancement of these cytoprotective mechanisms has potential value as a countermeasure to ocular alterations caused by radiation alone or in combination with other factors in spaceflight environments.

Zanello, Susana B.↗

Discovering Innovations in Stress Tolerance through Comparative Gene Regulatory Network Analysis and Cell-Type Specific Expression Maps (Final Technical Report with Cover Page)

Through this grant, we developed a comparative framework to elucidate the mechanisms behind variations in environmental stress responses among a diverse group of species within the Brassicaceae family. Our focus was on the differences in physiological and transcriptomic responses to abscisic acid (ABA), a hormone associated with water stress. We examined the differential growth responses of four Brassicaceae species, finding that most exhibited reduced root growth correlated with smaller meristem size. In contrast, Schrenkiella parvula showed accelerated growth due to increased root cell elongation. We employed RNA sequencing to analyze the transcriptional responses to ABA across these species, and innovative bioinformatics techniques were used to pinpoint biological pathways with significant divergence. Additionally, we utilized DAP-seq to map the gene regulatory networks associated with ABAresponsive transcription factors, revealing that variations in the regulation of growth hormone biosynthesis play a critical role in the distinct ABA effects on root growth among the species. This research sets a new standard for comparative physiology by integrating comparative genomics and transcriptomics to uncover pathway divergences.

59 BASIC BIOLOGICAL SCIENCES↗

Ambulation Increases Decompression Sickness in Altitude Exposure

INTRODUCTION - Exercise accelerates inert gas elimination during oxygen breathing prior to decompression (prebreathe), but may also promote bubble formation and increase the risk of decompression sickness (DCS). The timing, pattern and intensity of exercise are likely critical to the net effect. The NASA Prebreathe Reduction Program (PRP) combined oxygen prebreathe and exercise preceding a 4.3 psi exposure in non-ambulatory subjects (a microgravity analog) to produce two protocols now used by astronauts preparing for extravehicular activity (CEVIS and ISLE). Additional work is required to investigate whether exercise normal to 1 G environments increases the risk of DCS over microgravity simulation. METHODS - The CEVIS protocol was replicated with one exception. Our subjects completed controlled ambulation (walking in place with fixed cadence and step height) during both preflight and at 4.3 psi instead of remaining non-ambulatory throughout. Decompression stress was graded with aural Doppler (Spencer 0-IV scale). Two-dimensional echocardiographic imaging was used to look for left heart gas emboli (the presence of which prompted test termination). Venous blood was collected at three points to correlate Doppler measures of decompression stress with microparticle (cell fragment) accumulation. Fisher Exact Tests compared test and control groups. Trial suspension would occur when DCS risk >15% or grade IV venous gas emboli (VGE) risk >20% (at 70% confidence). RESULTS - Eleven person-trials were completed (9 male, 2 female) when DCS prompted suspension. DCS was greater than in CEVIS trials (3/11 [27%] vs. 0/45 [0%], respectively, p=0.03). Statistical significance was not reached for peak grade IV VGE (2/11 [18%] vs. 3/45 [7%], p=0.149) or cumulative grade IV VGE observations per subject-trial (8/128 [6%] vs. 26/630 [4%], p=0.151). Microparticle data were collected for 5/11 trials (3 with DCS outcomes), with widely varying patterns that could not be resolved statistically. CONCLUSION - We did find that that ambulation increases decompression stress. Additional trials would improve the statistical power to assess differences in VGE and to evaluate the relationship between decompression stress and microparticles.

Conkin, Johnny↗

Development of lithium diffused radiation resistant solar cells, part 2

The work performed to investigate the effect of various process parameters on the performance of lithium doped P/N solar cells is described. Effort was concentrated in four main areas: (1) the starting material, (2) the boron diffusion, (3) the lithium diffusion, and (4) the contact system. Investigation of starting material primarily involved comparison of crucible grown silicon (high oxygen content) and Lopex silicon (low oxygen content). In addition, the effect of varying growing parameters of crucible grown silicon on lithium cell output was also examined. The objective of the boron diffusion studies was to obtain a diffusion process which produced high efficiency cells with minimal silicon stressing and could be scaled up to process 100 or more cells per diffusion. Contact studies included investigating sintering of the TiAg contacts and evaluation of the contact integrity.

Payne, P. R.↗

Safety testing of lithium cells

Safety testing is intended to simulate, under laboratory conditions and controls, situations that will subject a cell to externally induced stress. The stresses can occur at any time during the useful life of the cell, from the time of manufacture until it is expended during mission deployment. Abuse testing can be divided into three major categories: Electrical, Mechanical, and Thermal. Although electrical abuses are generally found to occur during handling or deployment, Mechanical and Thermal stresses can be induced during transportation and storage. Therefore, it would be prudent to include predicted environmental exposure as part of the test plan. In the selection of a test program. specific test requirements should be tailored to meet the predicted mission requirements.

Liberto, Nick↗

Cell prestress. I. Stiffness and prestress are closely associated in adherent contractile cells

The tensegrity hypothesis holds that the cytoskeleton is a structure whose shape is stabilized predominantly by the tensile stresses borne by filamentous structures. Accordingly, cell stiffness must increase in proportion with the level of the tensile stress, which is called the prestress. Here we have tested that prediction in adherent human airway smooth muscle (HASM) cells. Traction microscopy was used to measure the distribution of contractile stresses arising at the interface between each cell and its substrate; this distribution is called the traction field. Because the traction field must be balanced by tensile stresses within the cell body, the prestress could be computed. Cell stiffness (G) was measured by oscillatory magnetic twisting cytometry. As the contractile state of the cell was modulated with graded concentrations of relaxing or contracting agonists (isoproterenol or histamine, respectively), the mean prestress ((t)) ranged from 350 to 1,900 Pa. Over that range, cell stiffness increased linearly with the prestress: G (Pa) = 0.18(t) + 92. While this association does not necessarily preclude other interpretations, it is the hallmark of systems that secure shape stability mainly through the prestress. Regardless of mechanism, these data establish a strong association between stiffness of HASM cells and the level of tensile stress within the cytoskeleton.

NASA Discipline Cell Biology↗

Circadian clock control of ribosome composition promotes rhythmic translation and termination fidelity

Ribosome composition is dynamic, shifting with cell state and stress, but whether it varies with circadian time is unknown. Here, we uncover circadian clock-driven changes in ribosome composition in Neurospora crassa . Mass spectrometry of ribosomes across circadian time identified six ribosomal proteins and one associated factor under clock control. Rhythms in eL31 abundance were validated in purified ribosomes, and deletion of el31 disrupted translation rhythms in nearly half of rhythmically translated mRNAs. N. crassa eL31 promotes circadian control of translation termination and impacts elongation fidelity while maintaining Mg homeostasis, a key determinant of translational accuracy. These findings reveal that the circadian clock reprograms ribosome composition to orchestrate rhythmic translation and fidelity, temporally expanding the proteome beyond the static genome to align cellular function with time of day.

circadian clock↗

Device for tensioning test specimens within an hermetically sealed chamber

The device is characterized by a support column adapted to be received within an insulated, hermetically sealable chamber. A plurality of anchor pins are mounted on the column for releasibly connecting thereto a plurality of test specimens. A plurality of axially displaceable pull rods are received by the column in coaxial alignment with the anchor pins. One end of each pull rod is provided with a coupling for connecting the pull rod to a test specimen. The opposite end of the pull rod is entended through a cover plate and adapted to be connected with a remotely related linear actuator through a connecting link including a load cell for measuring stress as the pull rod is placed in tension by the actuator.

Evans, P. K.↗

The scanning electron microscope as a tool in space biology

Normal erythrocytes are disc-shaped and are referred to here descriptively as discocytes. Several morphologically variant forms occur nomally but in rather small amounts, usually less than one percent of total. It has been shown though, that spiculed variant forms referred to as echinocytes are generated in significant amounts at zero g. Normal red cells have been stressed in vitro in an effort to duplicate the observed discocyte-echinocyte transformation at zero g. The significance of this transformation to extended stay in space and some of the plausible reasons for this transformation are discussed.

Barrett, R. A.↗