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Biosentinel: Developing a Space Radiation Biosensor

Ionizing radiation presents a major challenge to human exploration and long-term residence in space. The deep-space radiation spectrum includes highly energetic particles that generate double strand breaks (DSBs), deleterious DNA lesions that are usually repaired without errors via homologous recombination (HR), a conserved pathway in all eukaryotes. While progress identifying and characterizing biological radiation effects using Earth-based facilities has been significant, no terrestrial source duplicates the unique space radiation environment.We are developing a biosensor-based nanosatellite to fly aboard NASAs Space Launch System Exploration Mission 1, expected to launch in 2017 and reach a 1AU (astronomic unit) heliocentric orbit. Our biosensor (called BioSentinel) uses the yeast S. cerevisiae to measure DSBs in response to ambient space radiation. The BioSentinel strain contains engineered genetic defects that prevent growth until and unless a radiation-induced DSB near a reporter gene activates the yeasts HR repair mechanisms. Thus, culture growth and metabolic activity directly indicate a successful DSB-and-repair event. In parallel, HR-defective and wild type strains will provide survival data. Desiccated cells will be carried within independent culture microwells, built into 96-well microfluidic cards. Each microwell set will be activated by media addition at different time points over 18 months, and cell growth will be tracked continuously via optical density. One reserve set will be activated only in the occurrence of a solar particle event. Biological measurements will be compared to data provided by onboard physical dosimeters and to Earth-based experiments.BioSentinel will conduct the first study of biological response to space radiation outside Low Earth Orbit in over 40 years. BioSentinel will thus address strategic knowledge gaps related to the biological effects of space radiation and will provide an adaptable platform to perform human-relevant measurements in multiple space environments. We hope that it can therefore be used on the ISS, on and around other planetary bodies as well as other exploration platforms as a self-contained system that will allow us to compare and calibrate different radiation environments.BioSentinels results will be critical for improving interpretation of the effects of space radiation exposure, and for reducing the risk associated with long-term human exploration.

space radiation

BioSentinel

The BioSentinel mission was selected in 2013 as one of three secondary payloads to fly on the Space Launch Systems first Exploration Mission (EM-1) planned for launch in December 2017. The primary objective of BioSentinel is to develop a biosensor using a simple model organism to detect, measure, and correlate the impact of space radiation to living organisms overlong durations beyond Low Earth Orbit(LEO). While progress identifying and characterizing biological radiation effects using Earth-based facilities has been significant, no terrestrial source duplicates the unique space radiation environment.The BioSentinel biosensor uses the budding

Hanel, Robert

Phase Sensitive X-Ray Diffraction Imaging of Defects in Biological Macromolecular Crystals

Characterization of defects and/or disorder in biological macromolecular crystals presents much greater challenges than in conventional small-molecule crystals. The lack of sufficient contrast of defects is often a limiting factor in x-ray diffraction topography of protein crystals. This has seriously hampered efforts to understand mechanisms and origins of formation of imperfections, and the role of defects as essential entities in the bulk of macromolecular crystals. In this report, we employ a phase sensitive x-ray diffraction imaging approach for augmenting the contrast of defects in protein crystals.

Hu, Z. W.

Cosmo Cassette: A Microfluidic Microgravity Microbial System For Synthetic Biology Unit Tests and Satellite Missions

Although methods in the design-build-test life cycle of the synthetic biology field have grown rapidly, the expansion has been non-uniform. The design and build stages in development have seen innovations in the form of biological CAD and more efficient means for building DNA, RNA, and other biological constructs. The testing phase of the cycle remains in need of innovation. Presented will be both a theoretical abstraction of biological measurement and a practical demonstration of a microfluidics-based platform for characterizing synthetic biological phenomena. Such a platform demonstrates a design of additive manufacturing (3D printing) for construction of a microbial fuel cell (MFC) to be used in experiments carried out in space. First, the biocompatibility of the polypropylene chassis will be demonstrated. The novel MFCs will be cheaper, and faster to make and iterate through designs. The novel design will contain a manifold switchingdistribution system and an integrated in-chip set of reagent reservoirs fabricated via 3D printing. The automated nature of the 3D printing yields itself to higher resolution switching valves and leads to smaller sized payloads, lower cost, reduced power and a standardized platform for synthetic biology unit tests on Earth and in space. It will be demonstrated that the application of unit testing in synthetic biology will lead to the automatic construction and validation of desired constructs. Unit testing methodologies offer benefits of preemptive problem identification, change of facility, simplicity of integration, ease of documentation, and separation of interface from implementation, and automated design.

Bioelectrochemical System

Enhanced Monte Carlo Simulations for Electron Energy Loss Mitigation in Real-Space Nanoimaging of Thick Biological Samples and Microchips

High-resolution imaging using Transmission Electron Microscopy (TEM) is essential for applications such as grain boundary analysis, microchip defect characterization, and biological imaging. However, TEM images are often compromised by electron energy spread and other factors. In TEM mode, where the objective and projector lenses are positioned downstream of the sample, electron–sample interactions cause energy loss, which adversely impacts image quality and resolution. This study introduces a simulation tool to estimate the electron energy loss spectrum (EELS) as a function of sample thickness, covering electron beam energies from 300 keV to 3 MeV. Leveraging recent advances in MeV-TEM/STEM technology, which includes a state-of-the-art electron source with 2-picometer emittance, an energy spread of 3 × 10 -5 , and optimized beam characteristics, we aim to minimize energy spread. By integrating EELS capabilities into the BNL Monte Carlo (MC) simulation code for thicker samples, we evaluate electron beam parameters to mitigate energy spread resulting from electron–sample interactions. Based on our simulations, we propose an experimental procedure for quantitively distinguishing between elastic and inelastic scattering. The findings will guide the selection of optimal beam settings, thereby enhancing resolution for nanoimaging of thick biological samples and microchips.

36 MATERIALS SCIENCE

Life Sciences Accomplishments 1994

The NASA Life and Biomedical Sciences and Applications Division (LBSAD) serves the Nation's life sciences community by managing all aspects of U.S. space-related life sciences research and technology development. The activities of the Division are integral components of the Nation's overall biological sciences and biomedical research efforts. However, NASA's life sciences activities are unique, in that space flight affords the opportunity to study and characterize basic biological mechanisms in ways not possible on Earth. By utilizing access to space as a research tool, NASA advances fundamental knowledge of the way in which weightlessness, radiation, and other aspects of the space-flight environment interact with biological processes. This knowledge is applied to procedures and technologies that enable humans to live and work in and explore space and contributes to the health and well-being of people on Earth. The activities of the Division are guided by the following three goals: Goal 1) Use microgravity and other unique aspects of the space environment to enhance our understanding of fundamental biological processes. Goal 2) Develop the scientific and technological foundations for supporting exploration by enabling productive human presence in space for extended periods. Goal 3) Apply our unique mission personnel, facilities, and technology to improve education, the quality of life on Earth, and U.S. competitiveness. The Division pursues these goals with integrated ground and flight programs involving the participation of NASA field centers, industry, and universities, as well as interactions with other national agencies and NASA's international partners. The published work of Division-sponsored researchers is a record of completed research in pursuit of these goals. During 1993, the LBSAD instituted significant changes in its experiment solicitation and peer review processes. For the first time, a NASA Research Announcement (NRA) was released requesting proposals for ground-based and flight research for all programs. Areas of particular interest to NASA were defined Proposals due April 29, 1994, will be peer reviewed - externally for scientific merit. This annual NRA process is now the mechanism for recruiting both extramural and intramural investigations. As an overview of LBSAD activities in 1993, this accomplishments document covers each of the major organizational components of the Division and the accomplishments of each. The second section is a review of the Space Life Sciences Research programs Space Biology, Space Physiology and Countermeasures, Radiation Health, Environmental Health, Space Human Factors, Advanced Life Support, and Global Monitoring and Disease Prediction, The third section, Research in Space Flight, describes the substantial contributions of the Spacelab Life Sciences 2 (SLS-2) mission to life sciences research and the significant contributions of the other missions flown in 1993, along with plans for future missions. The Division has greatly expanded and given high priority to its Education and Outreach Programs, which are presented in the fourth section. The fifth and final section, Partners for Space, shows the Divisions Cooperative efforts with other national and international agencies to achieve common goals, along with the accomplishments of joint research and analysis programs.

Burnell, Mary Lou

The Impact of Deviation from Michaelis-Menten Saturation on Mathematical Model Stability Properties

Based on purely abstract ecological theory, it has been argued that a system composed of two or more consumers competing for the same resource cannot persist. By analysis on a Monod format mathematical model, Hubble and others demonstrated that this assertion is true for all but very special cases of such competing organisms which are determined by an index formed by a grouping of. the parameters which characterize the biological processes of the competing organisms. In the laboratory, using a bioreactor, Hansen and Hubble obtained confirmatory results for several cases of two competing species, and they characterized it as "qualitative confirmation" of the assertion. This result is amazing, since the analysis required the exact equality of the hey index, and it seems certain that no pair of organism species could have exactly equal values. It is quite plausible, however, that pairs of organism species could have approximately equal indices, and the question of how different they could be and still have coexistence of the two (or more) presents itself. In this paper, the pursuit of this question and a compatible resolution is presented.

Blackwell, Charles

BioSentinel: Mission Development of a Radiation Biosensor to Gauge DNA Damage and Repair Beyond Low Earth Orbit on a 6U Nanosatellite.

We are designing and developing a "6U" (10 x 22 x 34 cm; 14 kg) nanosatellite as a secondary payload to fly aboard NASA's Space Launch System (SLS) Exploration Mission (EM) 1, scheduled for launch in late 2017. For the first time in over forty years, direct experimental data from biological studies beyond low Earth orbit (LEO) will be obtained during BioSentinel's 12- to 18- month mission. BioSentinel will measure the damage and repair of DNA in a biological organism and allow us to compare that to information from onboard physical radiation sensors. In order to understand the relative contributions of the space environment's two dominant biological perturbations, reduced gravity and ionizing radiation, results from deep space will be directly compared to data obtained in LEO (on ISS) and on Earth. These data points will be available for validation of existing biological radiation damage and repair models, and for extrapolation to humans, to assist in mitigating risks during future long-term exploration missions beyond LEO. The BioSentinel Payload occupies 4U of the spacecraft and will utilize the monocellular eukaryotic organism Saccharomyces cerevisiae (yeast) to report DNA double-strand-break (DSB) events that result from ambient space radiation. DSB repair exhibits striking conservation of repair proteins from yeast to humans. Yeast was selected because of 1) its similarity to cells in higher organisms, 2) the well-established history of strains engineered to measure DSB repair, 3) its spaceflight heritage, and 4) the wealth of available ground and flight reference data. The S. cerevisiae flight strain will include engineered genetic defects to prevent growth and division until a radiation-induced DSB activates the yeast's DNA repair mechanisms. The triggered culture growth and metabolic activity directly indicate a DSB and its successful repair. The yeast will be carried in the dry state within the 1-atm P/L container in 18 separate fluidics cards with each card having 16 independent culture microwells, with integral microchannels and filters to supply nutrients and reagents, confine the yeast to the wells, and enable optical measurement. The measurement subsystem will monitor each subgroup of culture wells continuously for several weeks, optically tracking DSBtriggered cell growth and metabolism. BioSentinel will also include physical radiation sensors based on the TimePix sensor, as implemented by JSC's RadWorks group, which record individual radiation events including estimates of their linear-energytransfer (LET) values. Radiation-dose and LET data will be compared directly to the rate of DSB-and-repair events measured by the S. cerevisiae biosentinels. The spacecraft bus will operate in a deep space environment with functions that include command and data handling, communications, power generation (via deployable solar panels) and storage, and attitude determination-and-control system with micropropulsion. Development of the BioSentinel spacecraft will mature and prove multiple nanosatellite advances in order to function well beyond LEO: Communications from distances of ≥ 500,000 km; Autonomous attitude control, momentum management, and safe mode of nanosatellites in deep space; Shielding-, hardening-, design-, and software-derived radiation tolerance for electronics; Reliable functionality for 12 - 18 months of key subsystems for biofluidics, memory, communications, power, etc.; Close integration of living biological radiation event monitors with miniature physical radiation spectrometers; Biological measurement of solar particle events beyond Earth orbit In addition to providing the first biological results from beyond LEO in over 4 decades, BioSentinel will provide an adaptable small-satellite instrument platform to perform a range of human-exploration-relevant measurements that characterize the biological consequences of multiple outer space environments. BioSentinel is being developed under NASA's Advanced Exploration Systems program.

DNA damage

Computational Nanotechnology at NASA Ames Research Center, 1996

Some forms of nanotechnology appear to have enormous potential to improve aerospace and computer systems; computational nanotechnology, the design and simulation of programmable molecular machines, is crucial to progress. NASA Ames Research Center has begun a computational nanotechnology program including in-house work, external research grants, and grants of supercomputer time. Four goals have been established: (1) Simulate a hypothetical programmable molecular machine replicating itself and building other products. (2) Develop molecular manufacturing CAD (computer aided design) software and use it to design molecular manufacturing systems and products of aerospace interest, including computer components. (3) Characterize nanotechnologically accessible materials of aerospace interest. Such materials may have excellent strength and thermal properties. (4) Collaborate with experimentalists. Current in-house activities include: (1) Development of NanoDesign, software to design and simulate a nanotechnology based on functionalized fullerenes. Early work focuses on gears. (2) A design for high density atomically precise memory. (3) Design of nanotechnology systems based on biology. (4) Characterization of diamonoid mechanosynthetic pathways. (5) Studies of the laplacian of the electronic charge density to understand molecular structure and reactivity. (6) Studies of entropic effects during self-assembly. Characterization of properties of matter for clusters up to sizes exhibiting bulk properties. In addition, the NAS (NASA Advanced Supercomputing) supercomputer division sponsored a workshop on computational molecular nanotechnology on March 4-5, 1996 held at NASA Ames Research Center. Finally, collaborations with Bill Goddard at CalTech, Ralph Merkle at Xerox Parc, Don Brenner at NCSU (North Carolina State University), Tom McKendree at Hughes, and Todd Wipke at UCSC are underway.

Globus, Al

Fractals in biology and medicine

Our purpose is to describe some recent progress in applying fractal concepts to systems of relevance to biology and medicine. We review several biological systems characterized by fractal geometry, with a particular focus on the long-range power-law correlations found recently in DNA sequences containing noncoding material. Furthermore, we discuss the finding that the exponent alpha quantifying these long-range correlations ("fractal complexity") is smaller for coding than for noncoding sequences. We also discuss the application of fractal scaling analysis to the dynamics of heartbeat regulation, and report the recent finding that the normal heart is characterized by long-range "anticorrelations" which are absent in the diseased heart.

Review

Longitudinal Plasma Proteomic Profiling Reveals Divergent Immune Responses in Durably Cured and Relapsed Pulmonary Tuberculosis

Background: Predicting the risk of tuberculosis (TB) relapse is vital to improving treatment outcomes. Although clinical risk factors of relapse are well characterized, the biological mechanisms driving relapse, particularly host immune responses, remain poorly understood. Elucidating these mechanisms is necessary to better predict relapse risk. Methods: We conducted a longitudinal, global proteomic study on 60 participants with active pulmonary TB, half who were durably cured and half who relapsed. Plasma was collected at seven time-points: at treatment initiation (baseline), during therapy, and 52 weeks post-baseline. Samples were analyzed by high-resolution LC-MS/MS. Results: 2,418 proteins were identified across all samples, with 1,756 being differentially expressed relative to baseline (unadjusted p < 0.05). 956 proteins were differentially abundant between cured and relapsed participants. Relapsed participants showed heightened humoral immunity throughout treatment, as well as upregulated complement activation and HDL particles. Cured participants exhibited elevated recovery-related pathways by week 4, including downregulated epithelial invasion and upregulated oxygen transport. Conclusions: Heightened humoral and innate immune responses were associated with relapse, whereas recovery signatures were associated with durable cure. These findings advance our understanding of host responses to treatment and provide a basis for developing blood-based biomarkers to identify patients at increased risk of relapse.

LC-MS/MS

Signature analysis of high-throughput transcriptomics screening data for mechanistic inference and chemical grouping

Abstract High-throughput transcriptomics (HTTr) uses gene expression profiling to characterize the biological activity of chemicals in in vitro cell-based test systems. As an extension of a previous study testing 44 chemicals, HTTr was used to screen an additional 1,751 unique chemicals from the EPA’s ToxCast collection in MCF7 cells using 8 concentrations and an exposure duration of 6 h. We hypothesized that concentration-response modeling of signature scores could be used to identify putative molecular targets and cluster chemicals with similar bioactivity. Clustering and enrichment analyses were conducted based on signature catalog annotations and ToxPrint chemotypes to facilitate molecular target prediction and grouping of chemicals with similar bioactivity profiles. Enrichment analysis based on signature catalog annotation identified known mechanisms of action (MeOAs) associated with well-studied chemicals and generated putative MeOAs for other active chemicals. Chemicals with predicted MeOAs included those targeting estrogen receptor (ER), glucocorticoid receptor (GR), retinoic acid receptor (RAR), the NRF2/KEAP/ARE pathway, AP-1 activation, and others. Using reference chemicals for ER modulation, the study demonstrated that HTTr in MCF7 cells was able to stratify chemicals in terms of agonist potency, distinguish ER agonists from antagonists, and cluster chemicals with similar activities as predicted by the ToxCast ER Pathway model. Uniform manifold approximation and projection (UMAP) embedding of signature-level results identified novel ER modulators with no ToxCast ER Pathway model predictions. Finally, UMAP combined with ToxPrint chemotype enrichment was used to explore the biological activity of structurally related chemicals. The study demonstrates that HTTr can be used to inform chemical risk assessment by determining in vitro points of departure, predicting chemicals’ MeOA and grouping chemicals with similar bioactivity profiles.

Toxicology

Assessment of wetland productive capacity from a remote-sensing-based model - A NASA/NMFS joint research project

NASA and the U.S. National Marine Fisheries Service have undertaken the development of Landsat Thematic Mapper (TM) technology for the evaluation of the usefulness of wetlands to estuarine fish and shellfish production. Toward this end, a remote sensing-based Productive Capacity model has been developed which characterizes the biological and hydrographic features of a Gulf Coast Marsh to predict detrital export. Regression analyses of TM simulator data for wetland plant production estimation are noted to more accurately estimate the percent of total vegetative cover than biomass, indicating that a nonlinear relationship may be involved.

Butera, M. K.

Aircraft laser derived chlorophyll distribution across the Iceland-Faeroe front

The ocean surface color (water-leaving radiances) and thermal structure across the Iceland-Faeroe Front under both clear and cloudy conditions were measured from a NASA research aircraft on 25 May 1989. The measurements were made along four north-south lines that were 125 km in length and spaced 35 km apart. The color measurements were made with a 14 channel, non-polarized Multispectral Airborne Radiometer System (MARS) while the thermal data were collected by a thermal radiometer and aircraft bathythermographs. The satellite imagery (NOAA AVHRR) sequence show the development of meanders through the frontal region. These aircraft ocean color and thermal data characterize the biological distribution and are closely coupled to the physical processes occurring in the frontal systems. The ratio of several channels of the ocean color data are used to determine the surface chlorophyll. The retrieved data correlate well with laser-induced chlorophyll fluorescence obtained at the same time and historical chlorophyll data. The observed chlorophyll patchiness appearing across the Iceland-Faeroe Front is believed to be directly and indirectly related to primary and secondary circulation processes in ocean frontal systems. High chlorophyll concentrations were observed on the north side of the front and are inferred to be the result from the advection of Icelandic Coastal Water into the region. The sharp chlorophyll decline at and south of the thermal frontal boundary is clearly related to the subsurface thermal structure.

Arnone, Robert A.

New Chemical, Bio-Optical and Physical Observations of Upper Ocean Response to the Passage of a Mesoscale Eddy Off Bermuda

A mesoscale eddy advected across the Bermuda Testbed Mooring site over a 30-day period centered on July 14, 1995. Temperature and current measurements along with biogeochemical measurements were used to characterize the biological response of the upper ocean associated with the introduction of nitrate into the euphotic layer due to the doming of isotherms associated with the eddy. Complementary shipboard data showed an anomalous water mass, which extended from a depth of approximately 50 to 1000 m, manifesting as a cold surface expression and warm anomaly at depth. Although mesoscale eddies are frequently observed in the Sargasso Sea, the present observations are particularly unique because of the high-temporal-resolution measurements of the new instrumentation deployed on the mooring. Analyzers that measure nitrate plus nitrite were placed at depths of 80 and 200 m and bio-optical sensors were located at depths of 20, 35, 45, 71, and 86 m. Peak nitrate values of nearly 3.0 micro-M at 80 m and chlorophyll a values of 1.4 mg/cubic m at 71 m were observed, a well as a 25- to 30-meter shoaling of the 1% light level depth. A Doppler shift from the inertial period (22.8 hours) to 25.2 hours was observed in several time series records due to the movement of the eddy across the mooring. Inertial pumping brought cold, nutrient-rich waters farther into the euphotic zone than would occur solely by isothermal lifting. Silicic acid was depleted to undetectable levels owing to the growth of diatoms within the eddy. The chlorophyll a values associated with the eddy appear to be the largest recorded during the eight years of the ongoing US JGOFS Bermuda Atlantic Time Series Study program.

McNeil, J. D.

New Chemical, Bio-Optical and Physical Observations of Upper Ocean Response to the Passage of a Mesoscale Eddy off Bermuda

A mesoscale eddy advected across the Bermuda Testbed Mooring site over a 30-day period centered on July 14, 1995. Temperature and current measurements along with biogeochemical measurements were used to characterize the biological response of the upper ocean associated with the introduction of nitrate into the euphoric layer due to the doming of isotherms associated with the eddy. Complementary shipboard data showed an anomalous water mass, which extended from a depth of approximately 50 to 1000 m, manifesting as a cold surface expression and warm anomaly at depth. Although mesoscale eddies are frequently observed in the Sargasso Sea, the present observations are particularly unique because of the high-temporal-resolution measurements of the new instrumentation deployed on the mooring. Analyzers that measure nitrate plus nitrite were placed at depths of 80 and 200 m and bio-optical sensors were located at depths of 20, 35, 45, 71, and 86 m. Peak nitrate values of nearly 3.0 microns at 80 m and chlorophyll alpha values of 1.4 mg/cu m at 71 m were observed, as well as a 25- to 30-meter shoaling of the 1% light level depth. A Doppler shift from the inertial period (22.8 hours) to 25.2 hours was observed in several time series records due to the movement of the eddy across the mooring. Inertial pumping brought cold, nutrient-rich waters farther into the euphotic zone than would occur solely by isothermal lifting. Silicic acid was depleted to undetectable levels owing to the growth of diatoms within the eddy. The chlorophyll alpha values associated with the eddy appear to be the largest recorded during the 8 years of the ongoing U.S. JGOFS Bermuda Atlantic Time Series Study (BATS) program.

McNeil, J. D.

Short Time Scale Evolution of Microbiolites in Rapidly Receding Altiplanic Lakes: Learning How to Recognize Changing Signatures of Life

As part of the exploration of high altitude lakes as analogs to Martian paleolakes environment, we are investigating a remarkably large and diverse field of lacustrine stromatolites located at 4,365m in the Bolivian Altiplano (22 deg 47 00 min S and 67 deg 47.00 min W).The field is composed of both early Holocene fossil structures located on paleoshorelines and present-day active cyanobacterial communities on the shore and at the bottom of the current Laguna Blanca and Verde. Its physical environment, broad diversity of morphologies, and their associated spatial heterogeneity, origin, and scale offer a unique opportunity to explore microbiolites in conditions reminiscent of early Earth and Mars. At this altitude and latitude, UV radiation levels are enhanced (40% higher than sea level) and harmful to microorganisms living in shallow waters which provide only minimal protection from UV. Similar conditions prevailed on early Earth when the ozone layer had yet to be formed in the atmosphere. Compared to those studied at sea levels, these stromatolites could yield new insights about the earliest terrestrial forms of life. Moreover, the combination of physical and geological environment of this site is exceptionally analogous to conditions believed to be prevalent on Mars at the end of the Noachian (3.5 Ga ago), allowing to test the potential for forming stromatolites in martian paleolakes and learn how to identify their fossil record remotely. Our overarching goal is to generate new astrobiological information on high-altitude stromatolites as clues to early biospheres with implications for Earth and Mars. Our two central objectives are: (1) characterize the biological, geological, and mineralogical features and significance of this field, and to identify geo-signatures such as morphology, geology, chronostratigraphy, mineralogy and biosignatures, and (2) to facilitate remote-sensing and ground robotic detection capabilities for future astrobiological missions to Mars.

Cabrol, N. A.