Fruit Fly Lab - 01 Payload Overview

Presentation to POIWG meeting at MSFC to discuss planned operations for upcoming FFL-01 mission on SpaceX-5. Will show hardware suite used, on-orbit operations, training strategy, and data handling architecture

Securing tropical forest carbon: the contribution of protected areas to REDD

Forest loss and degradation in the tropics contribute 6-17% of all greenhouse gas emissions. Protected areas cover 217.2 million ha (19.6%) of the world's humid tropical forests and contain c. 70.3 petagrams of carbon (Pg C) in biomass and soil to 1 m depth. Between 2000 and 2005, we estimate that 1.75 million ha of forest were lost from protected areas in humid tropical forests, causing the emission of 0.25-0.33 Pg C. Protected areas lost about half as much carbon as the same area of unprotected forest. We estimate that the reduction of these carbon emissions from ongoing deforestation in protected sites in humid tropical forests could be valued at USD 6,200-7,400 million depending on the land use after clearance. This is >1.5 times the estimated spending on protected area management in these regions. Improving management of protected areas to retain forest cover better may be an important, although certainly not sufficient, component of an overall strategy for reducing emissions from deforestation and forest degradation (REDD

Development of Two Color Fluorescent Imager and Integrated Fluidic System for Nanosatellite Biology Applications

Nanosatellites offer frequent, low-cost space access as secondary payloads on launches of larger conventional satellites. We summarize the payload science and technology of the Microsatellite in-situ Space Technologies (MisST) nanosatellite for conducting automated biological experiments. The payload (two fused 10-cm cubes) includes 1) an integrated fluidics system that maintains organism viability and supports growth and 2) a fixed-focus imager with fluorescence and scattered-light imaging capabilities. The payload monitors temperature, pressure and relative humidity, and actively controls temperature. C. elegans (nematode, 50 m diameter x 1 mm long) was selected as a model organism due to previous space science experience, its completely sequenced genome, size, hardiness, and the variety of strains available. Three strains were chosen: two green GFP-tagged strains and one red tdTomato-tagged strain that label intestinal, nerve, and pharyngeal cells, respectively. The integrated fluidics system includes bioanalytical and reservoir modules. The former consists of four 150 L culture wells and a 4x5 mm imaging zone the latter includes two 8 mL fluid reservoirs for reagent and waste storage. The fluidic system is fabricated using multilayer polymer rapid prototyping: laser cutting, precision machining, die cutting, and pressure-sensitive adhesives it also includes eight solenoid-operated valves and one mini peristaltic pump. Young larval-state (L2) nematodes are loaded in C. elegans Maintenance Media (CeMM) in the bioanalytical module during pre-launch assembly. By the time orbit is established, the worms have grown to sufficient density to be imaged and are fed fresh CeMM. The strains are pumped sequentially into the imaging area, imaged, then pumped into waste. Reagent storage utilizes polymer bags under slight pressure to prevent bubble formation in wells or channels. The optical system images green and red fluorescence bands by excitation with blue (473 nm peak) and amber (587 nm peak) LEDs it achieves 8 m lateral resolution using a CMOS imaging chip (as configured for serial data speeds) or 4 m resolution using USB imaging chips. The imager consists of a modified commercial off-the-shelf CMOS chip camera, amber, blue and white LEDs, as well as a relay lens and dual-band filters to obviate moving parts while supporting both fluorescence wavelengths

Innate Immune Responses of Drosophila Melanogaster are Altered by Spaceflight

Alterations and impairment of immune responses in humans present a health risk for space exploration missions. The molecular mechanisms under pinning innate immune defense can be confounded by the complexity of the acquired immune system of humans. Drosophila (fruit fly) innate immunity is simpler, and shares many similarities with human innate immunity at the level of molecular and genetic pathways. The goals of this study were to elucidate fundamental immune processes in Drosophila affected by spaceflight and to measure host-pathogen responses post-flight. Five containers, each containing ten female and five male fruit flies, were housed and bred on the space shuttle (average orbit altitude of330.35 km) for 12 days and 18.5 hours. A new generation of flies was reared in microgravity. In larvae, the immune system was examined by analyzing plasmatocyte number and activity in culture. In adults, the induced immune responses were analyzed by bacterial clearance and quantitative real-time polymerase chain reaction (qPCR) of selected genes following infection with E. coli. The RNA levels of relevant immune pathway genes were determined in both larvae and adults by microarray analysis. The ability of larval plasmatocytes to phagocytose E. coli in culture was attenuated following spaceflight, and in parallel, the expression of genes involved in cell maturation was down regulated. In addition, the level of constitutive expression of pattern recognition receptors and opsonins that specifically recognize bacteria, and of lysozymes, antimicrobial peptide (AMP) pathway and immune stress genes, hallmarks of humoral immunity, were also reduced in larvae. In adults, the efficiency of bacterial clearance measured in vivo following a systemic infection with E. coli post-flight, remained robust. We show that spaceflight altered both cellular and humoral immune responses in Drosophila and that the disruption occurs at multiple interacting pathways

Spaceflight Causes Increased Virulence of Serratia Marcescens on a Drosophila Melanogaster Host

Drosophila melanogaster, or the fruit fly, has long been an important organism for Earth-based research, and is now increasingly utilized as a model system to understand the biological effects of spaceflight. Studies in Drosophila melanogaster have shown altered immune responses in 3rd instar larvae and adult males following spaceflight, changes similar to those observed in astronauts. In addition, spaceflight has also been shown to affect bacterial physiology, as evidenced by studies describing altered virulence of Salmonella typhimurium following spaceflight and variation in biofilm growth patterns for the opportunistic pathogen Pseudomonas aeruginosa during flight. We recently sent Serratia marcescens Db11, a Drosophila pathogen and an opportunistic human pathogen, to the ISS on SpaceX-5 (Fruit Fly Lab-01). S. marcescens samples were stored at 4degC for 24 days on-orbit and then allowed to grow for 120 hours at ambient station temperature before being returned to Earth. Upon return, bacteria were isolated and preserved in 50% glycerol or RNAlater. Storage, growth, and isolation for ground control samples were performed using the same procedures. Spaceflight and ground samples stored in 50% glycerol were diluted and injected into 5-7-day-old ground-born adult D. melanogaster. Lethality was significantly greater in flies injected with the spaceflight samples compared to those injected with ground bacterial samples. These results indicate a shift in the virulence profile of the spaceflight S. marcescens Db11 and will be further assessed with molecular biological analyses. Our findings strengthen the conclusion that spaceflight impacts the virulence of bacterial pathogens on model host organisms such as the fruit fly. This research was supported by NASA's ISS Program Office (ISSPO) and Space Life and Physical Sciences Research and Applications (SLPSRA)

EcAMSat: Small Satellite to Examine E. coli's Response in Microgravity to the Antibiotic Gentamicin

We have successfully flown the EcAMSat (Escherichia coli Antimicrobial Satellite) free-flyer mission. This was a 6U small satellite that autonomously conducted an experiment in low Earth orbit to explore the impact of the space environment on antibiotic resistance in uropathogenic E. coli (UPEC) and the role a particular sigma factor plays in the response. After being held in stasis during transport to orbit, two strains a wildtype UPEC and an isogenic mutant with a deleted gene that encodes a sigma factor were grown to stationary phase in a fluidic card inside EcAMSat's payload, then incubated with three concentrations of the antibiotic gentamicin. The payload then administered alamarBlue, a redox indicator, into all wells of the fluidic card. The cells were then incubated for 144 hours and metabolic activity was measured optically using the payloads' LED and detector system. Data were then telemetered to the ground and compared to a control experiment conducted in an identical satellite in a lab. The results of this experiment will help us better understand important therapeutic targets for treating bacterial infections on Earth and in space. Such targets are particularly relevant to deep-space and long-duration missions where crew may be more susceptible to infection and treatments for them may work differently

EcAMSat: A Small Satellite Flown to Explore the Role a Sigma Factor Plays in E. coli's Response to the Antibiotic Gentamicin

We have successfully flown the EcAMSat (Escherichia coli Antimicrobial Satellite) free-flyer mission. This was a 6U (six unit - CubeSat) small satellite that autonomously conducted an experiment in low Earth orbit to explore the impact of the space environment on antibiotic resistance in uropathogenic E. coli (UPEC) and the role a particular sigma factor plays in the response. After being held in stasis during transport to orbit, two strains - a wildtype UPEC and an isogenic mutant with a deleted gene that encodes a sigma factor - were grown to stationary phase in a fluidic card inside EcAMSat's payload, then incubated with three concentrations of the antibiotic gentamicin. The payload then administered alamarBlue (registered trademark), a redox indicator, into all wells of the fluidic card. The cells were then incubated for 144 hours and metabolic activity was measured optically using the payloads' LED (Light-Emitting Diode) and detector system. Data were then telemetered to the ground and compared to a control experiment conducted in an identical satellite in a lab. The results of this experiment will help us better understand important therapeutic targets for treating bacterial infections on Earth and in space. Such targets are particularly relevant to deep-space and long-duration missions where crew may be more susceptible to infection and treatments for them may work differently

Payload Hardware and Experimental Protocol for Testing the Effect of Space Microgravity on the Resistance to Gentamicin of Stationary-Phase Uropathogenic Escherichia Coli and Its Sigma (sup S)-Deficient Mutant

Human immune response is compromised and bacteria can become more antibiotic resistant in space microgravity (MG). We report that under low-shear modeled microgravity (LSMMG) stationary-phase uropathogenic Escherichia coli (UPEC) become more resistant to gentamicin (Gm). UPEC causes urinary tract infections (UTIs), reported to afflict astronauts; Gm is a standard treatment, so these findings could impact astronaut health. Because LSMMG has been shown to differ from MG, we report here preparations to examine UPEC's Gm sensitivity during spaceflight using the E. coli Anti-Microbial Satellite (EcAMSat) on a free flying nanosatellite in low Earth orbit. Within EcAMSats payload, a 48-microwell fluidic card contains and supports study of bacterial cultures at constant temperature; optical absorbance changes in cell suspensions are made at three wavelengths for each microwell and a fluid-delivery system provides growth medium and predefined Gm concentrations. Performance characterization is reported for spaceflight prototypes of this payload system. Using conventional microtiter plates, we show that Alamar Blue (AB) absorbance changes due to cellular metabolism accurately reflect E. coli viability changes: measuring AB absorbance onboard EcAMSat will enable telemetry of spaceflight data to Earth. Laboratory results using payload prototypes are consistent with wellplate and flask findings of differential sensitivity of UPEC and its delta rpoS strain to Gm. Space MG studies using EcAMSat should clarify inconsistencies from previous space experiments on bacterial antibiotic sensitivity. Further, if sigma (sup s) plays the same role in space MG as in LSMMG and Earth gravity, EcAMSat results would facilitate utilizing our previously developed terrestrial UTI countermeasures in astronauts

Optimizing Sparse RFI Prediction using Deep Learning

Radio Frequency Interference (RFI) is an ever-present limiting factor among radio telescopes even in the most remote observing locations. When looking to retain the maximum amount of sensitivity and reduce contamination for Epoch of Reionization studies, the identification and removal of RFI is especially important. In addition to improved RFI identification, we must also take into account computational efficiency of the RFI-Identification algorithm as radio interferometer arrays such as the Hydrogen Epoch of Reionization Array grow larger in number of receivers. To address this, we present a Deep Fully Convolutional Neural Network (DFCN) that is comprehensive in its use of interferometric data, where both amplitude and phase information are used jointly for identifying RFI. We train the network using simulated HERA visibilities containing mock RFI, yielding a known "ground truth" dataset for evaluating the accuracy of various RFI algorithms. Evaluation of the DFCN model is performed on observations from the 67 dish build-out, HERA-67, and achieves a data throughput of 1.6$\times 10^{5}$ HERA time-ordered 1024 channeled visibilities per hour per GPU. We determine that relative to an amplitude only network including visibility phase adds important adjacent time-frequency context which increases discrimination between RFI and Non-RFI. The inclusion of phase when predicting achieves a Recall of 0.81, Precision of 0.58, and $F_{2}$ score of 0.75 as applied to our HERA-67 observations.Comment: 11 pages, 7 figure