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At least 343 records · Page 19

Developing Model Benchtop Systems for Microbial Experimental Evolution

Understanding how microbes impact an ecosystem has improved through advances of molecular and genetic tools, but creating complex systems that emulate natural biology goes beyond current technology. In fact, many chemical, biological, and metabolic pathways of even model organisms are still poorly characterized. Even then, standard laboratory techniques for testing microbial impact on environmental change can have many drawbacks; they are time-consuming, labor intensive, and are at risk of contamination. By having an automated process, many of these problems can be reduced or even eliminated. We are developing a benchtop system that can run for long periods of time without the need for human intervention, involve multiple environmental stressors at once, perform real-time adjustments of stressor exposure based on current state of the population, and minimize contamination risks. Our prototype device allows operators to generate an analogue of real world micro-scale ecosystems that can be used to model the effects of disruptive environmental change on microbial ecosystems. It comprises of electronics, mechatronics, and fluidics based systems to control, measure, and evaluate the before and after state of microbial cultures from exposure to environmental stressors. Currently, it uses four parallel growth chambers to perform tests on liquid cultures. To measure the population state, optical sensors (LED/photodiode) are used. Its primary selection pressure is UV-C radiation, a well-studied stressor known for its cell- and DNA-damaging effects and as a mutagen. Future work will involve improving the current growth chambers, as well as implementing additional sensors and environmental stressors into the system. Full integration of multiple culture testing will allow inter-culture comparisons. Besides the temperature and OD sensors, other types of sensors can be integrated such as conductivity, biomass, pH, and dissolved gasses such as CO and O. Additional environmental stressor systems like temperature (extreme heat or cold), metal toxicity, and other forms of radiation will increase the scale and testing range.

Developing↗

Improving Satellite Compatible Microdevices to Study Biology in Space

The technology for biology in space lags far behind the gold standard for biological experiments on Earth. To remedy this disparity, the Rothschild lab works on proof of concept, prototyping, and developing of new sensors and devices to further the capabilities of biology research on satellites. One such device is the PowerCell Payload System. One goal for synthetic biology in aiding space travel and colonization is to genetically engineer living cells to produce biochemicals in space. However, such farming in space presupposes bacteria retain their functionality post-launch, bombarded by radiation, and without the 1G of Earth. Our questions is, does a co-culture of cyanobacteria and protein-synthesizing bacteria produce Earth-like yields of target proteins? Is the yield sensitive to variable gravitational forces? To answer these questions, a PowerCell Payload System will spend 1 year aboard the German Aerospace Center's Euglena and Combined Regenerative Organic-food Production In Space (Eu:CROPIS) mission satellite. The PowerCell system is a pair of two 48-well microfluidic cards, each well seeded with bacteria. The system integrates fluidic, thermal, optical, electronic, and control systems to germinate bacteria spores, then measure the protein synthesized for comparison to parallel experiments conducted on the Earth. In developing the PowerCell Payload, we gained insight into the shortcomings of biology experiments on satellites. To address these issues, we have started three new prototyping projects: 1) The development of an extremely stable and radiation resistant cell-free system, allowing for the construction of proteins utilizing only cell components instead of living cells. This can be lyophilized on a substrate, like paper. (2) Using paper as a microfluidic platform that is flexible, stable, cheap, and wicking. The capillary action eliminates the need for pumps, reducing volume, mass, and potential failing points. Electrodes can be printed on the paper to sense for biochemicals. (3) Developing a modular, semi-autonomous microfluidic device that can be easily adapted for a variety of common biological experiments. This versatility will allow for quicker and cheaper experimentation. These improvements to satellite experiment platforms have the potential to radically increase the return from NASA's biological and field studies with reduced development time, mass, and cost with increased robustness data and interpretation.

biology in spac↗

Enabling Experimental Evolution: Multi-Parameter Sensor System Integration into a Culture/Stressor Biofluidics System

Experimental evolution (EE) exposes microbial communities to ecological stressors, simulating dynamics up to near-extinction events. Combined with comparative sequencing and other molecular tools, such data can inform the genetic and other biological mechanisms underlying extremophile adaptation, and other observed effects. Automating this type of experiment using biofluidics can mitigate many traditional obstacles, including delays in assay results and environment adjustment and the need for many replicates. A first-generation device for automating EE procedures, the Automated Adaptive Directed Evolution Chamber (AADEC), was developed at NASA Ames. UV-C radiation was the stressor, an LED-photodiode array measured optical density, magnetic agitation and peristaltic pump systems ensured nutrient availability, and Arduino microcontrollers provided control. Escherichia coli in LB kanamycin media was used for testing and performance verification. A manual laboratory procedure with timed exposure to UV-C was performed to typify tolerance acquisition. Approximately a 106 factor increase in survival ratio was recorded over multiple iterations. Currently, a second-generation device is being developed integrating more real-time sensors: redox potential (ORP), indicating available/consumed metabolic energy; dissolved oxygen (DO), indicating aerobic/anaerobic growth; pH, indicating metabolic products; and electrical conductivity (EC), another indicator of metabolic products. The EC sensor system was constructed and calibrated in-house and matched commercial sensors in the required range. A Raspberry Pi computer automated the electrical system, allowing real-time data acquisition. The fluidics card was made of CNC-milled polycarbonate for biocompatibility. Each sensor parameter can also be used as a selection pressure alone or in combination with others to create extreme microbial environments. As a proof of concept, this work demonstrated sensor operation in one pair of growth-sensor chambers. It can be expanded to a multi-chamber system to enable inter-culture comparisons and multi-population studies. The prior Arduino system will be ported to the RPi system. Future stressors to be added include thermal, reactive oxygen species, and varying nutrient availability.

Govinda Raj, Chinmayee↗

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.

Matin, A. C.↗

BioSentinel/Mars: Interplanetary Space Radiation Biosensor Experiment in Martian Transit on Mars 2020

Despite significant progress understanding biological radiation effects via terrestrial studies, no terrestrial source duplicates space’s unique radiation environment. Furthermore, no biological experiments have been conducted beyond low Earth orbit since Apollo. Understanding space’s fundamental biological effects requires overcoming these limitations. The BioSentinel 4U payload, under development for flight aboard Exploration Mission-1, measures biological responses to deep space radiation. Traveling to more than 1AU from Earth, BioSentinel/EM-1 will record DNA double-strand breaks (DSBs) repaired using a pathway common to humans and BioSentinel’s bioengineered yeast model organism, responding to as few as one biologically repaired DSB. The BioSentinel/Mars 4U instrument (6-8kg; 5-8W; 0.2-1MB/week) would include eighteen 16-well biosensor fluidic cards, activated biweekly during Mars 2020’s cruise phase, to provide a dose-dependent rate of DSB/repair. The instrument, which includes solid-state sensors for total ionizing dose and linear-energy-transfer spectra, addresses MEPAG SKG-B3 by simultaneously measuring both spectra and biological effects of space radiation. Biological measurements are rendered reliable by independent replicate experiments. The spatio-temporal uniformity of interplanetary galactic cosmic radiation makes BioSentinel/EM1 and BioSentinel/Mars approximate replicates, except for any major differences in solar particle events. Results will be compared to Earth and ISS controls to characterize the radiation/reduced-gravity parameter space by its biological impact.

BioSentinel↗

Simulation of Sweep-Jet Flow Control, Single Jet and Full Vertical Tail

This work is a simulation technology demonstrator, of sweep jets used to suppress boundary layer separation and increase maximum achievable load coefficients. A sweep jet is a discrete Coanda jet that oscillates in the plane parallel to an aerodynamic surface. It injects mass and momentum in the approximate stream wise direction. It also generate turbulent eddies at the oscillation frequency, which are typically large relative to boundary layer turbulence, and which augmenting mixing across the boundary layer to attack flow separation. Simulations of a fluidic oscillator, the sweep jet emerging from the oscillator, and the suppression of boundary layer separation by an array of sweep jets are performed. Simulation results are compared to data from a dedicated CFD validation experiment of a single oscillator and its sweep jet, and from a study of a full-scale Boeing 757 vertical tail augmented with an array of sweep jets.2, 20 A critical step in the work is the development of realistic time-dependent sweep-jet in flow boundary conditions, derived from the results of the single-oscillator simulations, which create the sweep jets in the full-tail simulations. Simulations were performed using the Over flow CFD solver, with high-order spatial discretization and a range of turbulence modeling. Good results were obtained for all flows simulated, when suitable turbulence modeling was used.

flow control↗

Assessment of Mixer-Ejector Nozzle with Thermal Acoustic Shield for Jet Noise Reduction

A tendency for excessive exhaust jet mixing noise from low bypass ratio turbofan engines is recognized as a key challenge in the design of commercial supersonic aircraft. In this work we investigate a unique combination of two noise mitigation methods as a novel strategy to reduce jet mixing noise. First, a thermal acoustic shield (TAS) is used to reflect high frequency acoustic waves at small angles to the jet axis; second, a mixer-ejector (ME) nozzle is used to mechanically shield noise propagating at large angles to the axis. The ME shroud also provides a convenient location for a TAS nozzle and improves TAS effectiveness by limiting the downstream extent of high frequency noise generation. In an additional benefit for a velocity-matched TAS stream, the ME allows a reduction in strength of the TAS outer shear layer which could serve as a secondary noise source. The present work provides a quantitative assessment of the ME-TAS concept, using a combination of RANS CFD simulations, acoustic analogy calculations for the farfield Green's function, and surrogate-based modeling and parameter space exploration. We first evaluate a subscale configuration, then use scaling arguments to apply subscale results to the systems-level analysis of a flight configuration; the latter configuration includes a generic low bypass ratio turbofan engine with an engine-driven electric generator for supplementary heating of the TAS stream. Additional RANS CFD calculations are performed for a notional ME-TAS geometry based on the full scale configuration, and various modeling assumptions and operational characteristics are evaluated. The ME-TAS concept is shown to provide effective shielding for high frequency jet noise, and should enable comparable noise suppression to a stand-alone ME of considerably greater length, weight and drag. In addition to investigating the integrated ME-TAS system, the present work differs from previous research into TAS and related fluidic shield concepts through the inclusion of modern numerical analysis tools and the systematic numerical examination of various design parameters.

thermal acoustic shield↗

Surface Tension Containment Experiment (STCE) - Increasing Science Throughput on ISS

The Microgravity Science Glovebox (MSG) on the International Space Station (ISS) is used for fluid transfer in many types of experiments. Reagents are handled in the MSG to prevent their accidental release into the cabin. However, the MSG is currently over-subscribed, creating a backlog of users in flight. As a recourse, current experiments are underway to assess the possibility of moving certain science operations from the MSG into the open cabin of the ISS. The experiments are designed to assess the efficacy of exploiting surface tension as a control to prevent the unwanted release of liquids. Dyed water currently serves as an ersatz for potentially more hazardous liquids. Common wet-lab operations such as de/mating wetted Luer-Lok fittings, liquid-bearing container lid removal, and pipetting between well plates are performed illustrating the facility and challenges imposed by the microgravity environment. Concerning the latter, various pipette cannula sizes are deployed at various injection, withdrawal, and translations rates to map the existence, size, velocity, and trajectory of satellite droplets expected to form when breaking contact between the water surface and the pipette tip. Though such drops frequently form in terrestrial operations, they are nearly imperceptible and inconsequential—due in part to their speed and because gravity quickly returns them to the well plate from which they came. The use of airflow to capture and collect such satellite droplets is demonstrated. The dynamic stability of the liquid-filled well plates is quantified in response to a variety of crew-imparted disturbances. From a safety perspective, the results from the STCE are of immediate practical value. If such routine low-gravity capillary fluidic operations can be established as mundane, their performance may be moved out of the MSG and into the cabin, significantly increasing the efficiency of experiments performed on ISS.

Ungar, Eugene K.↗

Plant Water Management (PWM) Experiment

Plant Water Management is a technology demonstration of recent advances in micro-g capillary fluidics research applied to plant growth systems. It has applications in long-term food production systems for missions to the Moon and Mars, as well as the immediate need for ISS food supplements to the crew diet. PWM will demonstrate the low-gravity role of surface tension, wetting, and system geometry to effectively replace the role of gravity in certain terrestrial plant growth systems.

Hatch, Tyler↗

LTF/Fluid and Cryogenic Systems Branch

Functions of the Branch: Cross-Cutting Fluids Engineering; Research and Technology Development of Cryogenic Fluid Management (CFM) Systems; Component and System level; Design, Development Test and Evaluation (DDT&E) for Fluid, CFM, Propellant and Gaseous Systems; Fluid Component Specification and Selection; Code Compliance of Pressurized Systems; Fluid/Cryogenic System Performance Analysis and Trade Studies; Hazards Analysis and Risk Assessment of Fluidic Systems; Fluids Modeling and Simulation Capabilities

Fluid↗

Plant Water Management (PWM)

The NASA Plant Water Management technology demonstration exploits recent advances in microgravity capillary fluidics to develop techniques for plant growth in a space environment. Plant growth in the low-g environment is often hampered by inadequate aeration and over-saturation in the root zone. The present effort aims to make passive use of poorly wetting liquids (i.e., contaminated water) within unique system geometries that effectively replace the role of gravity in providing sufficient aeration and hydration for simulated plants. Several ISS demonstrations are in preparation for launch in 2019, including hydroponic and soil approaches. Supportive terrestrial and low-g drop tower tests are conducted to aid in experiment design via proof-of-concepts, limits of operation, system stability, and others. The implications to short and long term applications are discussed in relation to plant growth facilities for both near-term microgravity plant science research and long duration human exploration missions.

Hatch, Tyler↗

Building a Robust Commercial Microgravity Economy in Earth's Orbit: Economic Readiness Considerations

The reduced gravity environment of space provides a unique opportunity to further our understanding of various materials phenomena involving the molten, fluidic and gaseous states as well as life science applications where, contrary to earlier beliefs, microgravity induces changes in single cells and simple organisms; not only in large organisms with a complex overall response to gravity (or lack thereof). The potential breadth of commercial opportunities in microgravity thus spans over many verticals of the private sector with applications ranging from fiber optics, high-resolution crystals, microencapsulation, 3D organs to perfume and color dyes. Overall, products manufactured in microgravity hold the promise to have key properties surpassing their best terrestrial counterparts. Commercialization, also known as taking a new technology to market, is a journey in itself where the business, economic, market and technological components must align to generate a successful outcome. A business perspective is very different than technology maturation. In order for a technology to be ready for commercialization, it must not only be mature, but it must also have a compelling business case, and the means to scale up production must be identified and practical. Creating a robust economy in Earths orbit (Fig 1) is especially challenging because of the complexity (high risks, lack of standardization) involved in predicting future growth. This complexity can easily overwhelm the fact that many of the products have an attractive touch of space which aids with branding and marketing.This paper reviews the types of added value that can be extracted from space, with an emphasis on the microgravity environment. In addition, lessons learned from past commercialization efforts will be reviewed. While past efforts have yielded some point successes, they have as a whole failed to precipitate a sustainable LEO based market

Cozmuta, Ioana↗

Economic Readiness Level Considerations for a Robust LEO Economy

The reduced gravity environment of space provides a unique opportunity to further our understanding of various materials phenomena involving the molten, fluidic and gaseous states as well as life science applications where, contrary to earlier beliefs, microgravity induces changes in single cells and simple organisms; not only in large organisms with a complex overall response to gravity (or lack thereof). The potential breadth of commercial opportunities in microgravity thus spans over many verticals of the private sector with applications ranging from fiber optics, high-resolution crystals, microencapsulation, 3D organs to perfume and color dyes. Overall, products manufactured in microgravity hold the promise to have key properties surpassing their best terrestrial counterparts. Commercialization, also known as taking a new technology to market, is a journey in itself where the business, economic, market and technological components must align to generate a successful outcome. A business perspective is very different than technology maturation, which can be measured with the usual Technology Readiness Level (TRL) approach. In order for a technology to be ready for commercialization, it must not only be mature (high TRL), but it must also have a compelling business case, and the means to scale up production must be identified and practical. Creating a sustainable economy in an emerging market such as microgravity is especially challenging because of the complexity (high risks, lack of standardization) involved in predicting future growth. This complexity can easily overwhelm the fact that many of the products have an attractive touch of space which aids with branding and marketing.This paper builds upon the concept of the verticals of microgravity, capturing not only new lines of investigations but also promising killer apps originating from microgravity to discuss and define the notion of Economic Readiness Level (ERL). To advance in ERL, the technology itself may not necessarily need to mature at all, but the understanding of its economic potential does. Building upon ERL, a model that ultimately leads to the creation of pathways for infusion of private capital and a sustainable commercial microgravity LEO-Earth economy will be discussed.

Cozmuta, Ioana↗

Comparison of Mixing Characteristics for Several Fuel Injectors at Mach 8, 12, and 15 Hypervelocity Flow Conditions

CFD analysis is presented of the mixing characteristics and performance of three fuel injectors at flight Mach numbers of 8, 12, and 15. The Reynolds-averaged simulations (RAS) were carried out using the VULCAN-CFD solver. The high Mach number flow conditions match those of the experiments conducted as a part of the Enhanced Injection and Mixing Project (EIMP) at the NASA Langley Research Center. The EIMP aims to investigate scramjet fuel injection and mixing physics, improve the understanding of underlying physical processes, and develop enhancement strategies relevant to flight Mach numbers greater than 8. The injectors include a fuel placement device, a strut, and a fluidic vortical mixer, a ramp. These fuel injectors accomplish the necessary task of distributing and mixing fuel into the supersonic cross-flow, albeit via different strategies. For comparison, a flush-wall injector is also included. This type of injector generally represents the simplest method of introducing fuel into a scramjet combustor. The three injectors represent the baseline configurations of the EIMP experiments. The mixing parameters of interest, such as mixing efficiency and total pressure recovery, are computed from the RAS and compared for the three flight conditions and injector configurations. In addition to mixing efficiency and total pressure recovery, the combustion efficiency and thrust potential are also computed for the reacting simulations. Plotting the total pressure recovery and thrust potential as a function of mixing efficiency provides added insight into critical aspects of combustor performance as the flight condition and injector type are varied.

Shenoy, Rajiv R.↗

Gene Expression Measurement Module (GEMM) for space application: Design and validation

In order to facilitate studies on the impact of the space environment on biological systems, we have developed a prototype of GEMM (Gene Expression Measurement Module) - an automated, miniaturized, integrated fluidic system for in-situ measurements of gene expression in microbial samples. The GEMM instrument is capable of (1) lysing bacterial cell walls, (2) extracting and purifying RNA released from cells, (3) hybridizing the RNA to probes attached to a microarray and (4) providing electrochemical readout, all in a microfluidics cartridge. To function on small, uncrewed spacecraft, the conventional, laboratory protocols for both sample preparation and hybridization required significant modifications. Biological validation of the instrument was carried out on Synechococcus elongatus, a photosynthetic cyanobacterium known for its metabolic diversity and resilience to adverse conditions. It was demonstrated that GEMM yielded reliable, reproducible gene expression profiles. GEMM is the only high throughput instrument that can be deployed in near future on space platforms other than the ISS to advance biological research in space. It can also prove useful for numerous terrestrial applications in the field.

Kianoosh Peyvan↗

The Unrealized Potential of Superhydrophobic Substrates in Advanced Life Support Systems

Nearly all water processing equipment aboard spacecraft is to a large extent controlled by capillary forces arising from substrate wetting conditions. Superhydrophobic wetting conditions provide an essentially passive means to keep water away from certain substrates providing a level of no-moving-parts phase separation and control. This work presents a host of non-wetting aqueous microgravity capillary fluidics phenomena arising from interactions with easily fabricated superhydrophobic substrates. The value of such phenomena for potential life support applications aboard spacecraft is clear, especially for substrate properties that are thermally robust, corrosion resistant, and self-cleaning for both long- and short term applications. Large length scale low-g demonstrations of the phenomena are provided in HD video format for the extensive drop tower tests conducted. The broader crosscutting impacts to numerous fluids processing operations for life support are discussed. Current practical applications addressed in light of superhydrophobicity include urine-processing, water recovery, fire safety, and others.

Superhydrophoic↗

RDE Nozzle Computational Design Methodology Development and Application

The rotating detonation engine (RDE) is currently under investigation as an approach to achieving pressure gain combustion for propulsion and power systems. The RDE typically consists of an annulus with one end open (or having a throat and/or nozzle) and the other end valved using non-mechanical, fluidic means. In this study, a total of 6 nozzle design variations were studied for the NPS combustor geometry, using the validated quasi-2D to 3-D coupled simulation approach.

CFD↗

BioLEAD: BioLogical Exploration via Autonomous Detection – Using Dielectric Spectroscopy to Monitor Biological Changes in Response to Deep Space Radiation

Leveraging the bio-fluidic hardware developed for BioSentinel, we propose a new payload for autonomous missions: BioLEAD – BioLogical Exploration via Autonomous Detection. With NASA's renewed focus to continue human exploration of the Moon (Artemis Program), the BioLEAD payload aims to investigate the effects of the lunar environment on biology, whether onboard a lunar lander, the Lunar Gateway, or as a free-flying CubeSat orbiting the Moon. In place of an optical detection system, BioLEAD will employ a miniaturized, non-invasive dielectric spectroscopy sensor to enable real-time monitoring of biological activity. The sensor operates by relating capacitance measurements to the dielectric properties of the cell, such as cell morphology, doubling time, and cell cycle stage. The implementation of this new sensor technology will address limitations of the optical measurement system used on BioSentinel. It will also advance the use of autonomous bioanalytical microsystems and reveal new information regarding biological responses to the Moon’s radiation environment. Most importantly, BioLEAD’s technology will be adaptable for a wide array of future missions.

Biosensor↗