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Perfusion Bioreactor Module

Perfusion bioreactor module, self-contained, closed-loop cell-culture system that operates in microgravity or on Earth. Equipment supports growth or long-term maintenance of cultures of human or other fragile cells for experiments in basic cell biology or process technology. Designed to support proliferation (initially at exponential rates of growth) of cells in complex growth medium and to maintain confluent cells in defined medium under conditions optimized to permit or encourage selected functions of cells, including secretion of products of cells into medium.

Morrison, Dennis R.↗

Residual acceleration data on IML-1: Development of a data reduction and dissemination plan

The need to record some measure of the low-gravity environment of an orbiting space vehicle was recognized at an early stage of the U.S. Space Program. Such information was considered important for both the assessment of an astronaut's physical condition during and after space missions and the analysis of the fluid physics, materials processing, and biological sciences experiments run in space. Various measurement systems were developed and flown on space platforms beginning in the early 1970's. Similar in concept to land based seismometers that measure vibrations caused by earthquakes and explosions, accelerometers mounted on orbiting space vehicles measure vibrations in and of the vehicle due to internal and external sources, as well as vibrations in a sensor's relative acceleration with respect to the vehicle to which it is attached. The data collected over the years have helped to alter the perception of gravity on-board a space vehicle from the public's early concept of zero-gravity to the science community's evolution of thought from microgravity to milligravity to g-jitter or vibrational environment. Since the advent of the Shuttle Orbiter Program, especially since the start of Spacelab flights dedicated to scientific investigations, the interest in measuring the low-gravity environment in which experiments are run has increased. This interest led to the development and flight of numerous accelerometer systems dedicated to specific experiments. It also prompted the development of the NASA MSAD-sponsored Space Acceleration Measurement System (SAMS). The first SAMS units flew in the Spacelab on STS-40 in June 1991 in support of the first Spacelab Life Sciences mission (SLS-1). SAMS is currently manifested to fly on all future Spacelab missions.

Rogers, Melissa J. B.↗

Space medicine research: Needs for the 21st century

Space medicine research in the 21st century will continue to focus on the four major areas including: (1) expansion of the current incomplete knowledge base of clinical and subclinical physiological changes due to microgravity; (2) development of countermeasures to extend the capabilities of the human performance envelope in extended duration flights; (3) development of novel methods for delivering all aspects of a comprehensive health care system in extreme remote conditions: and (4) further research and application of systems for biological materials processing. New space transportation vehicles will place unique physiologic and human factors demands on the human system, while providing better access to platforms for materials processing. Success in meeting the demands in each of the noted research areas will require an extensive, interactive team approach. Personnel from the medical research,operational, developmental, and basic science communities will be essential to success.

Pepper, L. J.↗

Application of capillary fluid management techniques to the design of a phase separating microgravity bioreactor

Manned space missions require the development of compact, efficient, and reliable life support systems. A number of aqueous biological conversion processes are associated with bioregenerative life support systems. Vessels, or bioreactors, capable of supporting these processes in microgravity must be developed. An annular flow bioreactor has been conceived. It has the potential to incorporate containment, phase separation, gas exchange, and illumination into a single vessel. The bioreactor utilizes capillary fluid management techniques and is configured as a cylindrical tube in which a two-phase liquid-gas flow is maintained. Vanes placed around the inner perimeter enhance capillary forces and cause the liquid phase to attach and flow along the interior surface of the tube. No physical barrier is required to complete phase separation. It is shown analytically that liquid film thickness is limited only by vane geometry and that an annular flow bioreactor capable of managing 284 liters would occupy 0.7 cubic m, less than half the volume of a Spacelab experiment rack.

Finger, Barry W.↗

Protein Solvation in Membranes and at Water-Membrane Interfaces

Different salvation properties of water and membranes mediate a host of biologically important processes, such as folding, insertion into a lipid bilayer, associations and functions of membrane proteins. These processes will be discussed in several examples involving synthetic and natural peptides. In particular, a mechanism by which a helical peptide becomes inserted into a model membrane will be described. Further, the molecular mechanism of recognition and association of protein helical segments in membranes will be discussed. These processes are crucial for proper functioning of a cell. A membrane-spanning domain of glycophorin A, which exists as a helical dimer, serves as the model system. For this system, the free energy of dissociation of the helices is being determined for both the wild type and a mutant, in which dimerization is disrupted.

Pohorille, Andrew↗

Abstract of talk for Silicon Valley Linux Users Group

The use of Linux for research at NASA Ames is discussed.Topics include:work with the Atmospheric Physics branch on software for a spectrometer to be used in the CRYSTAL-FACE mission this summer; work on in the Neuroengineering Lab with code IC including an introduction to the extension of the human senses project,advantages with using linux for real-time biological data processing,algorithms utilized on a linux system, goals of the project,slides of people with Neuroscan caps on, and progress that has been made and how linux has helped.

Clanton, Sam↗

Evolution of catalytic function

An RNA-based evolution system was constructed in the laboratory and used to develop RNA enzymes with novel catalytic function. By controlling the nature of the catalytic task that the molecules must perform in order to survive, it is possible to direct the evolving population toward the expression of some desired catalytic behavior. More recently, this system has been coupled to an in vitro translation procedure, raising the possibility of evolving protein enzymes in the laboratory to produce novel proteins with desired catalytic properties. The aim of this line of research is to reduce darwinian evolution, the fundamental process of biology, to a laboratory procedure that can be made to operate in the service of organic synthesis.

NASA Program Exobiology↗

Effects of Convective Transport of Solute and Impurities on Defect-Causing Kinetics Instabilities in Protein Crystallization

Insight into the crystallization processes of biological macromolecules into crystals or aggregates can provide valuable guidelines in many fundamental and applied fields. Such insight will prompt new means to regulate protein phase transitions in-vivo, e.g., polymerization of hemoglobin S in the red cells, crystallization of crystallins in the eye lens, etc. Understanding of protein crystal nucleation will help achieve narrow crystallite size distributions, needed for sustained release of pharmaceutical protein preparations such as insulin or interferon. Traditionally, protein crystallization studies have been related to the pursuit of crystal perfection needed to improve the structure details provided by x-ray, electron or neutron diffraction methods. Crystallization trials for the purposes of structural biology carried out in space have posed an intriguing question related to the inconsistency of the effects of the microgravity growth on the quality of the crystals.

Vekilov, Peter G.↗

Three-Function Logic Gate Controlled by Analog Voltage

The figure is a schematic diagram of a complementary metal oxide/semiconductor (CMOS) electronic circuit that performs one of three different logic functions, depending on the level of an externally applied control voltage, V(sub sel). Specifically, the circuit acts as A NAND gate at V(sub sel) = 0.0 V, A wire (the output equals one of the inputs) at V(sub sel) = 1.0 V, or An AND gate at V(sub sel) = -1.8 V. [The nominal power-supply potential (VDD) and logic "1" potential of this circuit is 1.8 V.] Like other multifunctional circuits described in several prior NASA Tech Briefs articles, this circuit was synthesized following an automated evolutionary approach that is so named because it is modeled partly after the repetitive trial-and-error process of biological evolution. An evolved circuit can be tested by computational simulation and/or tested in real hardware, and the results of the test can provide guidance for refining the design through further iteration. The evolutionary synthesis of electronic circuits can now be implemented by means of a software package Genetic Algorithms for Circuit Synthesis (GACS) that was developed specifically for this purpose. GACS was used to synthesize the present trifunctional circuit. As in the cases of other multifunctional circuits described in several prior NASA Tech Briefs articles, the multiple functionality of this circuit, the use of a single control voltage to select the function, and the automated evolutionary approach to synthesis all contribute synergistically to a combination of features that are potentially advantageous for the further development of robust, multiple-function logic circuits, including, especially, field-programmable gate arrays (FPGAs). These advantages include the following: This circuit contains only 9 transistors about half the number of transistors that would be needed to obtain equivalent NAND/wire/AND functionality by use of components from a standard digital design library. If multifunctional gates like this circuit were used in the place of the configurable logic blocks of present commercial FPGAs, it would be possible to change the functions of the resulting digital systems within shorter times. For example, by changing a single control voltage, one could change the function of thousands of FPGA cells within nanoseconds. In contrast, typically, the reconfiguration in a conventional FPGA by use of bits downloaded from look-up tables via a digital bus takes microseconds.

Zebulum, Ricardo↗

Miniature Laboratory for Detecting Sparse Biomolecules

A miniature laboratory system has been proposed for use in the field to detect sparsely distributed biomolecules. By emphasizing concentration and sorting of specimens prior to detection, the underlying system concept would make it possible to attain high detection sensitivities without the need to develop ever more sensitive biosensors. The original purpose of the proposal is to aid the search for signs of life on a remote planet by enabling the detection of specimens as sparse as a few molecules or microbes in a large amount of soil, dust, rocks, water/ice, or other raw sample material. Some version of the system could prove useful on Earth for remote sensing of biological contamination, including agents of biological warfare. Processing in this system would begin with dissolution of the raw sample material in a sample-separation vessel. The solution in the vessel would contain floating microscopic magnetic beads coated with substances that could engage in chemical reactions with various target functional groups that are parts of target molecules. The chemical reactions would cause the targeted molecules to be captured on the surfaces of the beads. By use of a controlled magnetic field, the beads would be concentrated in a specified location in the vessel. Once the beads were thus concentrated, the rest of the solution would be discarded. This procedure would obviate the filtration steps and thereby also eliminate the filter-clogging difficulties of typical prior sample-concentration schemes. For ferrous dust/soil samples, the dissolution would be done first in a separate vessel before the solution is transferred to the microbead-containing vessel.

Lin, Ying↗

Understanding the Scalability of Bayesian Network Inference Using Clique Tree Growth Curves

One of the main approaches to performing computation in Bayesian networks (BNs) is clique tree clustering and propagation. The clique tree approach consists of propagation in a clique tree compiled from a Bayesian network, and while it was introduced in the 1980s, there is still a lack of understanding of how clique tree computation time depends on variations in BN size and structure. In this article, we improve this understanding by developing an approach to characterizing clique tree growth as a function of parameters that can be computed in polynomial time from BNs, specifically: (i) the ratio of the number of a BN s non-root nodes to the number of root nodes, and (ii) the expected number of moral edges in their moral graphs. Analytically, we partition the set of cliques in a clique tree into different sets, and introduce a growth curve for the total size of each set. For the special case of bipartite BNs, there are two sets and two growth curves, a mixed clique growth curve and a root clique growth curve. In experiments, where random bipartite BNs generated using the BPART algorithm are studied, we systematically increase the out-degree of the root nodes in bipartite Bayesian networks, by increasing the number of leaf nodes. Surprisingly, root clique growth is well-approximated by Gompertz growth curves, an S-shaped family of curves that has previously been used to describe growth processes in biology, medicine, and neuroscience. We believe that this research improves the understanding of the scaling behavior of clique tree clustering for a certain class of Bayesian networks; presents an aid for trade-off studies of clique tree clustering using growth curves; and ultimately provides a foundation for benchmarking and developing improved BN inference and machine learning algorithms.

Mengshoel, Ole J.↗

Observation Platform for Dynamic Biomedical and Biotechnology Experiments Using the International Space Station (ISS) Light Microscopy Module (LMM)

Techshot, Inc., has developed an observation platform for the LMM on the ISS that will enable biomedical and biotechnology experiments. The LMM Dynamic Stage consists of an electronics module and the first two of a planned suite of experiment modules. Specimens and reagent solutions can be injected into a small, hollow microscope slide-the heart of the innovation-via a combination of small reservoirs, pumps, and valves. A life science experiment module allows investigators to load up to two different fluids for on-orbit, real-time image cytometry. Fluids can be changed to initiate a process, fix biological samples, or retrieve suspended cells. A colloid science experiment module conducts microparticle and nanoparticle tests for investigation of colloid self-assembly phenomena. This module includes a hollow glass slide and heating elements for the creation of a thermal gradient from one end of the slide to the other. The electronics module supports both experiment modules and contains a unique illuminator/condenser for bright and dark field and phase contrast illumination, power supplies for two piezoelectric pumps, and controller boards for pumps and valves. This observation platform safely contains internal fluids and will greatly accelerate the research and development (R&D) cycle of numerous experiments, products, and services aboard the ISS.

Kurk, Michael A. (Andy)↗

Observation Platform for Dynamic Biomedical and Biotechnology Experiments Using the International Space Station (ISS) Light Microscopy Module (LMM)

Techshot, Inc., has developed an observation platform for the LMM on the ISS that will enable biomedical and biotechnology experiments. The LMM Dynamic Stage consists of an electronics module and the first two of a planned suite of experiment modules. Specimens and reagent solutions can be injected into a small, hollow microscope slide-the heart of the innovation-via a combination of small reservoirs, pumps, and valves. A life science experiment module allows investigators to load up to two different fluids for on-orbit, real-time image cytometry. Fluids can be changed to initiate a process, fix biological samples, or retrieve suspended cells. A colloid science experiment module conducts microparticle and nanoparticle tests for investigation of colloid self-assembly phenomena. This module includes a hollow glass slide and heating elements for the creation of a thermal gradient from one end of the slide to the other. The electronics module supports both experiment modules and contains a unique illuminator/condenser for bright and dark field and phase contrast illumination, power supplies for two piezoelectric pumps, and controller boards for pumps and valves. This observation platform safely contains internal fluids and will greatly accelerate the research and development (R&D) cycle of numerous experiments, products, and services aboard the ISS.

Kurk, Michael A. (Andy)↗

Monitoring Astronaut Health with DNA Sequencing

In recent years microbe a plethora of microbe populations have been identified onboard the ISS (International Space Station). Approaches for real-time tracking of microbes for routine housekeeping and food/water safety monitoring will be critical for mission safety and crew health on future longer duration missions to the Moon or Mars. This work is a proof-of-concept study demonstrating an end-to-end phylogenetic identification and full genome sequencing effort of multiple microbial populations. Our methodology utilized the ISS flight-certified WetLab-2 molecular toolbox and the Biomolecule Sequencer projects for real-time end-to-end on-orbit microbial biological samples processing and molecular analysis with real time results generated utilizing only field "offline" analytic software. For this experiment we colony-cultured several ISS isolated microorganisms before generation of the pre-sequencing library via the automated VolTRAX device which enabled high library turnover with little wet-bench activity or potential future costly astronaut time. The pre-sequencing library is diluted in loading buffer and injected into the MinION sample port, drawn into the nanopore window by capillary action, and sequenced using the MinKnown. 16S and full genome alignment, nucleotide matching, gene identification, and phylogenetic sorting was accomplished utilizing the Epi2me software and the offline NCBI Blast viral, microbiome, and human somatic databases. In short, the methodologies developed herein replace the myriad of specific, often highly targeted microbiological tests used in the clinical laboratory, which would be difficult if not impossible to currently implement aboard the ISS or in deep space, with a single metagenomics test.

genomics↗

Novel End-to-End Molecular Biology Approach for Direct Nanopore 1D cDNA Sequencing of Reverse Transcribed mRNAs Purified from Cell Cultures by the NASA ISS WetLab2 SPM

Continued space bioscience research onboard the International Space Station (ISS) and future long-duration flight missions to the Moon or Mars will require the ability to conduct on-orbit molecular analysis of biological samples independently from Earth. In the last year two new molecular analytic technologies have been installed and the technologies demonstrated onboard the ISS: The Sample Prep Module (SPM) WetLab-2 (WL2) qRT-PCR toolbox and the Oxford Nanopore MinIon Biomolecule Sequencer. Here we describe protocol development and integration into existing ISS technology for end-to-end on-orbit biological sample processing and molecular analysis with real time results generated utilizing only field offline analytic software. For this experiment we isolated primary cells from bone marrow flushes of wild type B6129SF2 mice (Jackson Labs) long bones. The cell isolate was then processed using the SPM to produce total 147nanograms of RNA. The total RNA was purified to only messenger RNA (mRNA) and transferred to Smartcycler Thermocycle ISS kit consumable tube using Eppendorf gel loading pipette tips for further processing. Complementary first strand cDNA was synthesized using OLIGO dT priming followed by addition of SuperScript II Reverse Transcriptase and thermal cycling as per manufacturers instruction. All thermal cycling was conducted using the ISS WetLab-2 Cephid Smarcycler real time thermal cycler. Our protocol takes advantage of mRNAs native poly(A) tail, synthesized in vivo to protect the mRNA from degradation by endonucleases, to eliminate end-prep for adapter ligation. The adapted library is purified using MyOne C1 Streptavidin beads before elution in buffer. The pre-sequencing library is diluted in the loading buffer and injected into the MinIon sample port, drawn into the nanopore window by capillary action, and sequenced using the MinKnown software with local basecalling. The sequencing read produced 34.5 million events and local basecalling produced 117,301 successful reads. NCBI Blast of the data for the mouse genome resulted in 2,462 successful nucleotide collection matches (gene sequences) exceeding 70 homology. These results demonstrate the viability of this novel flight ready end-to-end sample analytic methodology and provide a real time homolog for flight experimentation utilizing supply kits and technologies that have already been demonstrated on ISS.

MinIon↗

Delineation of vulnerable areas to water erosion in a mountain region using SDR-InVEST model: A case study of the Ourika watershed, Morocco

The Ourika watershed is characterized by higher than average flood flows, a rugged topography, a sparse vegetation cover and a friable substrate that makes it vulnerable to soil erosion. Soil is an important non-renewable asset capital, providing several ecosystem services that sustain life through physical and biological regulation processes. The loss, or otherwise, redistribution of biota by soil erosion in the Ourika watershed has received little attention and thus constitutes a key knowledge gap that needs to be addressed. This study quantifies the level of erosion and maps its spatial distribution in the watershed using the Sediment Delivery Ratio (SDR) of the Integrated Valuation of Ecosystem Services and Tradeoffs model (InVEST3.6.0). The SDR maps the overland sediment generation and delivery to the stream at the watershed level using rainfall, soil properties, topography, cover management and cropping practices as predefined set of input variables. Recommended internal model calibration was used and results were compared to similar studies carried out in different parts of Morocco for validation. The results reveal that 93% of the watershed is subject to a strong rainfall aggressiveness greater than 50 MJ mm ha–1 h–1 yr1 under an average rainfall intensity of 541 mm/year, rainfall capacity to initiate the detachment and transport of soil particles by runoff, and only 11.5% of the watershed area is protected with vegetative cover. We find that overall, the Ourika watershed is losing its soil to erosion at an alarming average rate of 258.48 ton.ha–1.yr–1, a rate well above the tolerable threshold of 7ton.ha–1.yr–1. The spatial distribution of this loss shows areas that need to be urgently remedied. The primary factors for soil erosion in the Ourika watershed appear to be related to the aggressive nature of local rainfall and the inherent susceptibility of soils, which can be costly to this cash-crop region. Because rainfall intensity, soil properties, and topography are intrinsic characteristics of the watershed, we suggest that seasonally adapted cropping rotations that maintains conservative practices with a high cover management may reduce soil erosion in the Ourika watershed.

Erosion↗

Hydrothermal Liquefaction of Wastewater Treatment Plant Solids

Feedstock cost is the greatest barrier to the commercial production of biofuels. The merits of any thermochemical or biological conversion process are constrained by their applicability to the lowest cost feedstocks. At PNNL, a recent resource assessment of wet waste feedstocks led to the identification of waste water treatment plant (WWTP) solids as a cost-negative source of biomass. WWTP solids disposal is a growing environmental concern [1, 2] and can account for up to half of WWTP operating costs. The high moisture content is well-suited for hydrothermal liquefaction (HTL), avoiding the costs and parasitic energy losses associated with drying the feedstock for incineration. The yield and quality of biocrude and upgraded biocrude from WWTP solids is comparable to that obtained from algae feedstocks but the feedstock cost is $500-1200 less per dry ton. A collaborative project was initiated and directed by the Water Environment & Reuse Foundation (WERF) and included feedstock identification, dewatering, shipping to PNNL, conversion to biocrude by HTL, and catalytic hydrothermal gasification of the aqueous byproduct. Additional testing at PNNL included biocrude upgrading by catalytic hydrotreatment, characterization of the hydrotreated product, and a preliminary techno-economic analysis (TEA) based on empirical results. This short article will cover HTL conversion and biocrude upgrading. The WERF project report with complete HTL results is now available through the WERF website [3]. The preliminary TEA is available as a PNNL report [4].

biocrude upgrading↗

Parallel Asynchronous Distributed-Memory Maximal Independent Set Algorithm with Work Ordering

The maximal independent set (MIS) graph problem arises in many applications such as computer vision, information theory, molecular biology, and process scheduling. The growing scale of graph data suggests the use of distributed memory hardware as a cost-effective approach to providing necessary compute and memory resources. Existing distributed memory parallel MIS algorithms rely on synchronous communication and use techniques such as subgraph computations. In this paper, we present an asynchronous distributed-memory parallel graph algorithm that relies on a virtual directed acyclic graph (DAG) that is created during the algorithm execution. We introduce two additional algorithms that save computations by ordering generated work. The first algorithm applies ordering globally to reduce computations, and the second algorithm applies ordering locally at the level of threads to minimize the synchronization overhead. We use two different implementations of Luby’s algorithm variants as baseline to compare the performance of the presented algorithms: (1) vertex-centric Luby A and Luby B implementations, and (2) the CombBLAS linear-algebra Luby A implementation. Results show that proposed algorithms outperform both implementations of Luby algorithms, especially in distributed execution. Furthermore, we show that for low-diameter graphs the algorithm that applies global ordering scales better than other algorithms and for high diameter graphs the original asynchronous algorithm and thread-level ordering algorithm show better performance.

maximal independent set (MIS), distributed memory ↗