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Search indexed NASA NTRS and DOE OSTI research on propulsion, heat transfer, battery materials and energy systems. Follow report and document links to the original sources.

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Culture-Independent Fungal Profiling for the International Space Station using Nanopore Sequencing: Method Development

Microbial monitoring of the International Space Station (ISS) environment is a crew health requirement that encompasses both bacterial and fungal identification. To achieve this currently, culture-based methods are used for sample collection, and these samples must be returned to the laboratory for analysis. The use of culture and the need for sample return to Earth results in a bias toward culturable organisms and causes a significant delay between sample collection and delivery of final data (weeks to months), respectively. Recently, advancements in molecular technology have aided a broad range of applications, including medical, industrial, and basic sciences. Additionally, increases in portability and ease-of-use of molecular platforms have provided point-of-use capabilities demonstrated by the miniPCR thermal cycler (miniPCR bio) and the MinION sequencer (Oxford Nanopore Technologies). Together, these devices have been applied to, and validated for, the identification of bacteria onboard the ISS. Building on this work, we have developed a spaceflight-compatible fungal workflow. Molecular-based fungal analysis is complicated by low biomass, difficult-to-lyse spores, debate regarding the region for taxonomic assignment, and the lack of bioinformatic pipelines and reference databases. To overcome these difficulties, primers yielding an ~ 2 Kb amplicon were validated against a wide range of ISS fungal isolates. The current spaceflight library preparation was substantially optimized, a bioinformatic pipeline was created, and refinements to the UNITE database were implemented. To compare this optimized method to the current culture-based standard, 30 sample sets (60 total swabs, two swabs held in tandem) were evaluated. Parallel fungal profiles were obtained between the two methods, with the culture-independent method revealing increased diversity. The addition of this method to the already established bacterial process fulfills the crew health identification requirement. Moreover, the implementation of this method onboard ISS will enhance our understanding of its unique fungal microbiome.

Hang N. Nguyen↗
NASA NTRS1987

What happens to the large eddies when net drag reduction is achieved by outer flow manipulators?

The tandem-arranged parallel plate manipulator (TAPPM) presently used to generate a flow with net drag reduction is conditionally sampled and found to exhibit significant changes of the boundary layer's large-scale motions at 20 delta(0), where delta(0) is the boundary layer thickness at the upstream plate of the TAPPM. Flow fields in the 'valleys' are equally affected. Both turbulent and irrotational flows then reestablish themselves by 50 delta(0). Flow visualization indicates that the TAPPM wake is very coherent at 20 delta(0), and has not yet spread into the wall region, while large scale motions and the ambient response flow continue to exhibit dynamic changes. This indicates that the shielding effect of the TAPPM's wake prolongs suppression of large-scale motions, thereby reducing skin friction over a sufficiently long extent of the boundary layer to overcome device drag and yield net drag reductions.

Falco, R. E.↗
NASA NTRS1989

Solid state lasers for use in non-contact temperature measurements

The last decade has seen a series of dramatic developments in solid state laser technology. Prominent among these has been the emergence of high power semiconductor laser diode arrays and a deepening understanding of the dynamics of solid state lasers. Taken in tandem these two developments enable the design of laser diode pumped solid state lasers. Pumping solid state lasers with semiconductor diodes relieves the need for cumbersome and inefficient flashlamps and results in an efficient and stable laser with the compactness and reliability. It provides a laser source that can be reliably used in space. These new coherent sources are incorporated into the non-contact measurement of temperature. The primary focus is the development and characterization of new optical materials for use in active remote sensors of the atmosphere. In the course of this effort several new materials and new concepts were studied which can be used for other sensor applications. The general approach to the problem of new non-contact temperature measurements has had two components. The first component centers on passive sensors using optical fibers; an optical fiber temperature sensor for the drop tube was designed and tested at the Marshall Space Flight Center. Work on this problem has given insight into the use of optical fibers, especially new IR fibers, in thermal metrology. The second component of the effort is to utilize the experience gained in the study of passive sensors to examine new active sensor concepts. By active sensor are defined as a sensing device or mechanism which is interrogated in some way be radiation, usually from a laser. The status of solid state lasers as sources for active non-contact temperature sensors are summarized. Some specific electro-optic techniques are described which are applicable to the sensor problems at hand. Work on some of these ideas is in progress while other concepts are still being worked out.

Buoncristiani, A. M.↗
NASA NTRS2017

Novel Approach to Quantification of Telomere Length with Direct Nanopore Sequencing and PCR Amplification

The ends of human chromosomes contain telomeres, or tandem arrays of repeating DNA sequences capped by multiple associated proteins that protect chromosomal ends from degradation. Telomeres function to preserve genomic stability by preventing natural chromosomal ends from being recognized as broken DNA double-strand breaks and triggering inappropriate DNA damage responses. Mounting evidence shows telomere length is an inherited trait that decreases with cellular division and normal aging. In addition, telomere length also appears to be influenced by other factors such as cellular oxidative stress, radiation and mechanical unloading of tissues as in microgravity. To measure these potential effects of the space environment on telomere lengths and cellular aging and regenerative potential we developed a novel telomere measurement approach based on nanopore sequencing of PCR amplified bar-coded chromosome termini. Specifically, telomeres can be directly enriched using barcode sequences ligated to the end of a free end- repaired telomere using the WetLab-2 facility SmartCycler on ISS. Prior to the ligation and amplification protocol a proteinase K digestion of capping proteins followed by a single 95-degree C heat denaturation of the protease is included. After digestion and bar-code ligation, PCR amplification will initiate with the ligated barcoded sequence, suppressing amplification of intra-genomic fragments and resulting in long read barcoded telomere amplicons including the nanopore motor protein sequences. Purified PCR amplicons are then used for nanopore sequencing library generation by simple addition of motor proteins and sequencing library is loaded into the MinION nanopore DNA-sequencer. Amplicon sequence reads from the nanopore device can be base-called quickly on ISS due to barcoding ligation and subsequent PCR amplification enhancing the telomere sequence resolution. If successfully implemented on ISS this technique will provide a novel means of measuring regenerative ability of somatic stem cells in astronauts, and of determining whether spaceflight in microgravity alters their telomere lengths and causes premature cellular aging.

Ma, Kristin R.↗