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NASA NTRS2003

Microsample Characterization of Coatings for Grcop-84 for High Heat Flux Applications

A multidisciplinary Johns Hopkins University-NASA Glenn team is undertaking a collaborative research program to elucidate and model the thermal stability and mechanical integrity of candidate coatings for GRCop-84. GRCop-84 is a high conductivity, high strength copper alloy that was recently developed at NASA Glenn for use in high temperature, high heat flux applications. With potential applications in rocket motor combustion chamber liners, nozzle ramps and other actively cooled structures, this new material offers great potential for decreasing weight and increasing reliability of third generation reusable launch vehicles. Current emphasis has turned toward the development of environmentally resistant and thermal barrier coatings for this alloy. Metallic coatings such as NiCrAlY and Cu-8-30%Cr have shown promise in: prohibiting blanching, reducing dog-house failures, increasing operating temperatures and decreasing cooling requirements. The focus of this research program is to develop a fundamental understanding of the substrate-coating interactions that occur during thermal cycling (inter-diffusion, viscoplasticity, morphological evolution, crack formation, etc.) and to derive a science-based protocol for future coating selection, optimization and reliability assurance. The microsample tensile testing approach adopted for this study allows us to characterize small-scale and highly scale-specific coatings and properties in a way not possible by conventional means. In addition to providing much needed design data, the integration of microsample testing with detailed microstructural observations provides a mechanistic foundation for coating optimization and life prediction modeling.

Hemker, Kevin↗
NASA NTRS2003

An On-Line Technology Information System (OTIS) for Advanced Life Support

An On-line Technology Information System (OTIS) is currently being developed for the Advanced Life Support (ALS) Program. This paper describes the preliminary development of OTIS, which is a system designed to provide centralized collection and organization of technology information. The lack of thorough, reliable and easily understood technology information is a major obstacle in effective assessment of technology development progress, trade studies, metric calculations, and technology selection for integrated testing. OTIS will provide a formalized, well-organized protocol to communicate thorough, accurate, current and relevant technology information between the hands-on technology developer and the ALS Community. The need for this type of information transfer system within the Solid Waste Management (SWM) element was recently identified and addressed. A SWM Technology Information Form (TIF) was developed specifically for collecting detailed technology information in the area of SWM. In the TIF, information is requested from SWM technology developers, based upon the Technology Readiness Level (TRL). Basic information is requested for low-TRL technologies, and more detailed information is requested as the TRL of the technology increases. A comparable form is also being developed for the wastewater processing element. In the future, similar forms will also be developed for the ALS elements of air revitalization, food processing, biomass production and thermal control. These ALS element-specific forms will be implemented in OTIS via a web-accessible interface,with the data stored in an object-oriented relational database (created in MySQLTM) located on a secure server at NASA Ames Research Center. With OTIS, ALS element leads and managers will be able to carry out informed research and development investment, thereby promoting technology through the TRL scale. OTIS will also allow analysts to make accurate evaluations of technology options. Additionally, the range and specificity of information solicited will help educate technology developers of programmatic needs.

Levri, Julie A.↗
NASA NTRS2009

Isokinetic Strength and Endurance Tests used Pre- and Post-Spaceflight: Test-Retest Reliability

To assess changes in muscular strength and endurance after microgravity exposure, NASA measures isokinetic strength and endurance across multiple sessions before and after long-duration space flight. Accurate interpretation of pre- and post-flight measures depends upon the reliability of each measure. The purpose of this study was to evaluate the test-retest reliability of the NASA International Space Station (ISS) isokinetic protocol. Twenty-four healthy subjects (12 M/12 F, 32.0 +/- 5.6 years) volunteered to participate. Isokinetic knee, ankle, and trunk flexion and extension strength as well as endurance of the knee flexors and extensors were measured using a Cybex NORM isokinetic dynamometer. The first weekly session was considered a familiarization session. Data were collected and analyzed for weeks 2-4. Repeated measures analysis of variance (alpha=0.05) was used to identify weekly differences in isokinetic measures. Test-retest reliability was evaluated by intraclass correlation coefficients (ICC) (3,1). No significant differences were found between weeks in any of the strength measures and the reliability of the strength measures were all considered excellent (ICC greater than 0.9), except for concentric ankle dorsi-flexion (ICC=0.67). Although a significant difference was noted in weekly endurance measures of knee extension (p less than 0.01), the reliability of endurance measure by week were considered excellent for knee flexion (ICC=0.97) and knee extension (ICC=0.96). Except for concentric ankle dorsi-flexion, the isokinetic strength and endurance measures are highly reliable when following the NASA ISS protocol. This protocol should allow accurate interpretation isokinetic data even with a small number of crew members.

Laughlin, Mitzi S.↗
NASA NTRS2010

On-Orbit Prospective Echocardiography on International Space Station

A number of echocardiographic research projects and experiments have been flown on almost every space vehicle since 1970, but validation of standard methods and the determination of Space Normal cardiac function has not been reported to date. Advanced Diagnostics in Microgravity (ADUM) -remote guided echocardiographic technique provides a novel and effective approach to on-board assessment of cardiac physiology and structure using a just-in-time training algorithm and real-time remote guidance aboard the International Space Station (ISS). The validation of remotely guided echocardiographic techniques provides the procedures and protocols to perform scientific and clinical echocardiography on the ISS and the Moon. The objectives of this study were: 1.To confirm the ability of non-physician astronaut/cosmonaut crewmembers to perform clinically relevant remotely guided echocardiography using the Human Research Facility on board the ISS. 2.To compare the preflight, postflight and in-flight echocardiographic parameters commonly used in clinical medicine.

Hamilton, Douglas R.↗
NASA NTRS2012

Molecular Technique to Reduce PCR Bias for Deeper Understanding of Microbial Diversity

Current planetary protection policies require that spacecraft targeted to sensitive solar system bodies be assembled and readied for launch in controlled cleanroom environments. A better understanding of the distribution and frequency at which high-risk contaminant microbes are encountered on spacecraft surfaces would significantly aid in assessing the threat of forward contamination. However, despite a growing understanding of the diverse microbial populations present in cleanrooms, less abundant microbial populations are probably not adequately taken into account due to technological limitations. This novel approach encompasses a wide spectrum of microbial species and will represent the true picture of spacecraft cleanroom-associated microbial diversity. All of the current microbial diversity assessment techniques are based on an initial PCR amplification step. However, a number of factors are known to bias PCR amplification and jeopardize the true representation of bacterial diversity. PCR amplification of a minor template appears to be suppressed by the amplification of a more abundant template. It is widely acknowledged among environmental molecular microbiologists that genetic biosignatures identified from an environment only represent the most dominant populations. The technological bottleneck overlooks the presence of the less abundant minority population and may underestimate their role in the ecosystem maintenance. DNA intercalating agents such as propidium monoazide (PMA) covalently bind with DNA molecules upon photolysis using visible light, and make it unavailable for DNA polymerase enzyme during polymerase chain reaction (PCR). Environmental DNA samples will be treated with suboptimum PMA concentration, enough to intercalate with 90 99% of the total DNA. The probability of PMA binding with DNA from abundant bacterial species will be much higher than binding with DNA from less abundant species. This will increase the relative DNA concentration of previously "shadowed" less abundant species available for PCR amplification. These PCR products obtained with and without PMA treatment will then be subjected to downstream diversity analyses such as sequencing and DNA microarray. It is expected that PMA-coupled PCR will amplify the "minority population" and help in understanding microbial diversity spectrum of an environmental sample at a much deeper level. This new protocol aims to overcome the major potential biases faced when analyzing microbial 16S rRNA gene diversity. This study will lead to a technological advancement and a commercial product that will aid microbial ecologists in understanding microbial diversity from various environmental niches. Implementation of this technique may lead to discoveries of novel microbes and their functions in sustenance of the ecosystem.

Vaishampayan, Parag A.↗
NASA NTRS1990

Performance of gigabit FDDI

Great interest exists in developing high speed protocols which will be able to support data rates at gigabit speeds. Hardware currently exists which can experimentally transmit at data rates exceeding a gigabit per second, but it is not clear as to what types of protocols will provide the best performance. One possibility is to examine current protocols and their extensibility to these speeds. Scaling of Fiber Distributed Data Interface (FDDI) to gigabit speeds is studied. More specifically, delay statistics are included to provide insight as to which parameters (network length, packet length or number of nodes) have the greatest effect on performance.

Game, David↗
NASA NTRS1994

Implementing bright light treatment for MSFC payload operations shiftworkers

Intense light can phase-shift circadian rhythms and improve performance, sleep, and wellbeing during shiftwork simulations, but to date there have been very few attempts to administer light treatment to real shiftworkers. We have developed procedures for implementing light treatment and have conducted controlled trials of light treatment for MSFC Payload Operations staff during the USML-1 mission. We found that treatment had beneficial effects on fatigue, alertness, self-rated job performance, sleep, mood, and work attendance. Although there are portable bright light boxes commercially available, there is no testing protocol and little performance information available. We measure the illuminance of two candidate boxes for use in this study and found that levels were consistently lower than those advertised by manufacturers. A device was developed to enhance the illuminance output of such units. This device increased the illuminance by at least 60 % and provided additional improvements in visual comfort and overall exposure. Both the design of this device and some suggested procedures for evaluating light devices are presented.

Hayes, Benita C.↗
NASA NTRS1995

An empirical evaluation of graphical interfaces to support flight planning

Whether optimization techniques or expert systems technologies are used, the underlying inference processes and the model or knowledge base for a computerized problem-solving system are likely to be incomplete for any given complex, real-world task. To deal with the resultant brittleness, it has been suggested that 'cooperative' rather than 'automated' problem-solving systems be designed. Such cooperative systems are proposed to explicitly enhance the collaboration of people and the computer system when working in partnership to solve problems. This study evaluates the impact of alternative design concepts on the performance of airline pilots interacting with such a cooperative system designed to support enroute flight planning. Thirty pilots were studied using three different versions of the system. The results clearly demonstrate that different system design concepts can strongly influence the cognitive processes of users. Indeed, one of the designs studied caused four times as many pilots to accept a poor flight amendment. Based on think-aloud protocols, cognitive models are proposed to account for how features of the computer system interacted with specific types of scenarios to influence exploration and decision-making by the pilots. The results are then used to develop recommendations for guiding the design of cooperative systems.

Smith, Philip J.↗
NASA NTRS2012

Environmental Control and Life Support (ECLS) Hardware Commonality for Exploration Vehicles

In August 2011, the Environmental Control and Life Support Systems (ECLSS) technical community, along with associated stakeholders, held a workshop to review NASA s plans for Exploration missions and vehicles with two objectives: revisit the Exploration Atmospheres Working Group (EAWG) findings from 2006, and discuss preliminary ECLSS architecture concepts and technology choices for Exploration vehicles, identifying areas for potential common hardware or technologies to be utilized. Key considerations for selection of vehicle design total pressure and percent oxygen include operational concepts for extravehicular activity (EVA) and prebreathe protocols, materials flammability, and controllability within pressure and oxygen ranges. New data for these areas since the 2006 study were presented and discussed, and the community reached consensus on conclusions and recommendations for target design pressures for each Exploration vehicle concept. For the commonality study, the workshop identified many areas of potential commonality across the Exploration vehicles as well as with heritage International Space Station (ISS) and Shuttle hardware. Of the 36 ECLSS functions reviewed, 16 were considered to have strong potential for commonality, 13 were considered to have some potential commonality, and 7 were considered to have limited potential for commonality due to unique requirements or lack of sufficient heritage hardware. These findings, which will be utilized in architecture studies and budget exercises going forward, are presented in detail.

Carrasquillo, Robyn↗
NASA NTRS2014

The Effects of Spaceflight and a Spaceflight Analog on Neurocognitive Perfonnance: Extent, Longevity, and Neural Bases

We are conducting ongoing experiments in which we are performing structural and functional magnetic resonance brain imaging to identify the relationships between changes in neurocognitive function and neural structural alterations following a six month International Space Station mission and following 70 days exposure to a spaceflight analog, head down tilt bedrest. Our central hypothesis is that measures of brain structure, function, and network integrity will change from pre to post intervention (spaceflight, bedrest). Moreover, we predict that these changes will correlate with indices of cognitive, sensory, and motor function in a neuroanatomically selective fashion. Our interdisciplinary approach utilizes cutting edge neuroimaging techniques and a broad ranging battery of sensory, motor, and cognitive assessments that will be conducted pre flight, during flight, and post flight to investigate potential neuroplastic and maladaptive brain changes in crewmembers following long-duration spaceflight. Success in this endeavor would 1) result in identification of the underlying neural mechanisms and operational risks of spaceflight-induced changes in behavior, and 2) identify whether a return to normative behavioral function following re-adaptation to Earth's gravitational environment is associated with a restitution of brain structure and function or instead is supported by substitution with compensatory brain processes. With the bedrest study, we will be able to determine the neural and neurocognitive effects of extended duration unloading, reduced sensory inputs, and increased cephalic fluid distribution. This will enable us to parse out the multiple mechanisms contributing to any spaceflight-induced neural structural and behavioral changes that we observe in the flight study. In this presentation I will discuss preliminary results from six participants who have undergone the bed rest protocol. These individuals show decrements in balance and functional mobility, and alterations in brain structure and function, in association with extended bed rest.

Seidler, R. D.↗
NASA NTRS2015

Evalution of a DE-Identification Process for Ocular Imaging

Medical privacy of NASA astronauts requires an organized and comprehensive approach when data are being made available outside NASA systems. A combination of factors, including the uniquely small patient population, the extensive medical testing done on these individuals, and the relative cultural popularity of the astronauts puts them at a far greater risk to potential exposure of personal information than the general public. Therefore, care must be taken to ensure that the astronauts' identities are concealed. Magnetic Resonance Imaging (MRI) medical data is a recent source of interest to researchers concerned with the development of Visual Impairment due to Intracranial Pressure (VIIP) in the astronaut population. Each vision MRI scan of an astronaut includes 176 separate sagittal images that are saved as an "image series" for clinical use. In addition to the medical information these image sets provide, they also inherently contain a substantial amount of non-medical personally identifiable information (PII) such as-name, date of birth, and date of exam. We have shown that an image set of this type can be rendered, using free software, to give an accurate representation of the patient's face. This currently restricts NASA from dispensing MRI data to researchers in a deidentified format. Automated software programs, such as the Brain Extraction Tool, are available to researchers who wish to de-identify MRI sagittal brain images by "erasing" identifying characteristics such as the nose and jaw on the image sets. However, this software is not useful to NASA for vision research because it removes the portion of the images around the eye orbits, which is the main area of interest to researchers studying the VIIP syndrome. The Lifetime Surveillance of Astronaut Health program has resolved this issue by developing a protocol to de-identify MRI sagittal brain images using Showcase Premier, a DICOM (Digital Imaging and Communications in Medicine) software package. The software allows manual editing of one image from a patient's image set to be automatically applied to the entire image series. This new approach would allow a new level of access to untapped medical imaging data relating to VIIP that can be utilized by researchers while protecting the privacy of the astronauts. In the next step toward finalizing this technique, NASA clinical radiology consultants will test the images to verify removal of all metadata and PII.

LaPelusa, Michael B.↗
NASA NTRSDate not supplied

Role of NDE and In-Situ Process Monitoring in Managing Risk of AM Space Hardware

The recently published NASA-STD-6030 defines the Additive Manufacturing (AM) Requirements for Spaceflight Systems. Key aspects of the certification approach include the development of a qualified material process (QMP) and material characterization determined by part classification. Nondestructive evaluation (NDE) of the full surface and volume is required for all part classifications except those with negligible risk. NASA is exploring the use of in-process monitoring data to improve risk posture and supplement post-build inspection for complex parts. Currently, the most challenging obstacle to overcome is linking the indications in the monitoring data to the physics of the process and the final material state of the finished part. NASA is undertaking studies to understand and quantify this relationship for various monitoring methods. The desired goal is to develop a protocol to establish this correlation for any monitoring method. Once this correlation is known, the critical defect size can be linked to a representative indication in the monitoring data, and the capability of the monitoring system can be tested using the 90/95 probability of detection requirement for NDE methods. This would enable the use of in-process monitoring as a defect screening activity for AM part certification. Many high-criticality components built with AM have high complexity and therefore limited inspectability, so using in-process monitoring can help address this certification gap. The use of adaptive, closed-loop monitoring systems that alter the locked process will require a new approach to the QMP.

additive manufacturing↗
NASA NTRSDate not supplied

Role of NDE and In-Situ Process Monitoring in Managing Risk of AM Space Hardware

The recently published NASA-STD-6030 defines the Additive Manufacturing (AM) Requirements for Spaceflight Systems. Key aspects of the certification approach include the development of a qualified material process (QMP) and material characterization determined by part classification. Nondestructive evaluation (NDE) of the full surface and volume is required for all part classifications except those with negligible risk. NASA is exploring the use of in-process monitoring data to improve risk posture and supplement post-build inspection for complex parts. Currently, the most challenging obstacle to overcome is linking the indications in the monitoring data to the physics of the process and the final material state of the finished part. NASA is undertaking studies to understand and quantify this relationship for various monitoring methods. The desired goal is to develop a protocol to establish this correlation for any monitoring method. Once this correlation is known, the critical defect size can be linked to a representative indication in the monitoring data, and the capability of the monitoring system can be tested using the 90/95 probability of detection requirement for NDE methods. This would enable the use of in-process monitoring as a defect screening activity for AM part certification. Many high-criticality components built with AM have high complexity and therefore limited inspectability, so using in-process monitoring can help address this certification gap. The use of adaptive, closed-loop monitoring systems that alter the locked process will require a new approach to the QMP.

advanced manufacturing↗
NASA NTRSDate not supplied

A Remote, Human-in-the-Loop Evaluation of a Multiple-Drone Delivery Operation

Over time, advances in unmanned aircraft systems (UAS) have enabled a shift in the operational paradigm from one operator managing one aircraft to that of multiple operators working together to manage multiple aircraft. This shift has highlighted the need for effective human-autonomy teaming methods to maintain manageable workload levels for operators as well as high standards of system performance and safety. This paper presents a study aimed at evaluating whether automation can help operators manage workload during small UAS (sUAS) package delivery scenarios featuring contingency situations. These contingency situations, resulting from unplanned UAS Volume Reservations (UVRs), required flight path reroutes for multiple aircraft simultaneously. The study manipulated the number of aircraft affected by the UVRs and the level of automation support. The presence of terrain conflicts was also controlled within each scenario. Due to the COVID-19 pandemic, subjects were not able to gain direct access to the Ground Control System (GCS). Therefore, the study was conducted using a subject-surrogate paradigm that required subjects to relay commands through a verbal protocol from remote locations outside of the lab to a researcher surrogate who had direct control of the GCS interfaces at the lab location. Results show that the automated support condition was associated with faster reroute response times, more efficient reroute maneuvers, and significantly lower levels of perceived workload than the manual reroute condition. However, the automation support level did not significantly impact pilots’ ability to avoid the UVR successfully; pilots were overwhelmingly capable of avoiding the UVR in all conditions. The presence of terrain conflicts primarily impacted pilot performance by leading to multiple uploads per vehicle, which was not typically required when pilots only needed to maneuver laterally. Although subjects did not have direct control over the GCS, subjective ratings indicate that the displays under test provided them with sufficient information to manage their aircraft and promptly respond to the unplanned UVRs. Overall, the objective and subjective data strongly suggest that the verbal protocol and subject-surrogate paradigm were effective methods for collecting data remotely amid the COVID-19 pandemic.

multi-UAS↗
NASA NTRS2003

The Application of Advanced Cultivation Techniques in the Long Term Maintenance of Space Flight Plant Biological Systems

The development of the International Space Station (ISS) presents extensive opportunities for the implementation of long duration space life sciences studies. Continued attention has been placed in the development of plant growth chamber facilities capable of supporting the cultivation of plants in space flight microgravity conditions. The success of these facilities is largely dependent on their capacity to support the various growth requirements of test plant species. The cultivation requirements for higher plant species are generally complex, requiring specific levels of illumination, temperature, humidity, water, nutrients, and gas composition in order to achieve normal physiological growth and development. The supply of water, nutrients, and oxygen to the plant root system is a factor, which has proven to be particularly challenging in a microgravity space flight environment. The resolution of this issue is particularly important for the more intensive crop cultivation of plants envisaged in Nasa's advanced life support initiative. BioServe Space Technologies is a NASA, Research Partnership Center (RPC) at the University of Colorado, Boulder. BioServe has designed and operated various space flight plant habitat systems, and placed specific emphasis on the development and enhanced performance of subsystem components such as water and nutrient delivery, illumination, gas exchange and atmosphere control, temperature and humidity control. The further development and application of these subsystems to next generation habitats is of significant benefit and contribution towards the development of both the Space Plant biology and the Advanced Life Support Programs. The cooperative agreement between NASA Ames Research center and BioServe was established to support the further implementation of advanced cultivation techniques and protocols to plant habitat systems being coordinated at NASA Ames Research Center. Emphasis was placed on the implementation of passive-based water and nutrient support systems and techniques, which can be used to minimize demands on power, mass, and operational complexity in space flight studies. This effort has direct application to the development of next-generation space flight plant chambers such as the Plant Research Unit (PRU). Work was also directed at the development of in-flight plant preservation techniques and protocols consistent with the interest in applying recent developments in gene chip micro array technologies. Cultivation technologies and protocols were evaluated in a 55 day space flight plant growth study, conducted on the ISS, mission 9A (10/7/02 - 12/7/02).

Heyenga, A. G.↗
NASA NTRS2004

Enhanced Characterization of Microorganisms in the Spacecraft Environment

Spacecraft such as the International Space Station (ISS) and the space shuttles are enclosed environments where crewmembers may spend long periods of time. Currently, crewmembers spend approximately a period of 6 months in the ISS. It is known that these prolonged stays in space may result in weakening of the immune system. Therefore, exposure to opportunistic pathogens or high concentrations of environmental microorganisms may compromise the health of the crew. The detection of biocontaminants in spacecraft environments utilizes culture-based methodology, omitting greater than 90% of all microorganisms including pathogens such as Legionella and Cryptosporidium. Culturable bacteria and fungi have been the only allergens studied; the more potent allergens, such as those from dust mites, have never been tested for in spacecraft environments. In addition, no attempts have been made to monitor microbial toxins in spacecrafts. The present study utilized quantitative polymerase chain reaction (QPCR) as a novel approach for monitoring microorganisms in the spacecraft environment. QPCR is a molecular biology technique that does not rely on the physiological state of the organisms for identification, thereby enabling detection of both culturable and non-culturable organisms. In this project, specific molecular primers and probes were utilized for the detection and quantitation of two fungi of concern in indoor environments, Aspergillus fumigatus and Stachybotrys chartarum. These organisms were selected because of the availability of PCR primers and probes, and to establish the sample processing and analysis methodology that may be employed with additional organisms. Purification methods and QPCR assays were optimized for the detection of these organisms in air, surface, and water; and sample processing and analysis protocols were developed. Preliminary validation of these protocols was conducted in the laboratory with air, surface, and water samples seeded with known concentrations of the target organisms. Additional studies were conducted with bulk materials (HEPA filter pleats and particulate found on the filter screen) obtained from the ISS.

Cruz, Patricia↗
NASA NTRS2002

Contrast and harmonic imaging improves accuracy and efficiency of novice readers for dobutamine stress echocardiography

BACKGROUND: Newer contrast agents as well as tissue harmonic imaging enhance left ventricular (LV) endocardial border delineation, and therefore, improve LV wall-motion analysis. Interpretation of dobutamine stress echocardiography is observer-dependent and requires experience. This study was performed to evaluate whether these new imaging modalities would improve endocardial visualization and enhance accuracy and efficiency of the inexperienced reader interpreting dobutamine stress echocardiography. METHODS AND RESULTS: Twenty-nine consecutive patients with known or suspected coronary artery disease underwent dobutamine stress echocardiography. Both fundamental (2.5 MHZ) and harmonic (1.7 and 3.5 MHZ) mode images were obtained in four standard views at rest and at peak stress during a standard dobutamine infusion stress protocol. Following the noncontrast images, Optison was administered intravenously in bolus (0.5-3.0 ml), and fundamental and harmonic images were obtained. The dobutamine echocardiography studies were reviewed by one experienced and one inexperienced echocardiographer. LV segments were graded for image quality and function. Time for interpretation also was recorded. Contrast with harmonic imaging improved the diagnostic concordance of the novice reader to the expert reader by 7.1%, 7.5%, and 12.6% (P < 0.001) as compared with harmonic imaging, fundamental imaging, and fundamental imaging with contrast, respectively. For the novice reader, reading time was reduced by 47%, 55%, and 58% (P < 0.005) as compared with the time needed for fundamental, fundamental contrast, and harmonic modes, respectively. With harmonic imaging, the image quality score was 4.6% higher (P < 0.001) than for fundamental imaging. Image quality scores were not significantly different for noncontrast and contrast images. CONCLUSION: Harmonic imaging with contrast significantly improves the accuracy and efficiency of the novice dobutamine stress echocardiography reader. The use of harmonic imaging reduces the frequency of nondiagnostic wall segments.

Non-NASA Center↗
NASA NTRSDate not supplied

Optimizing Single Nuclei Sequencing of Brain Samples From Space Flown Mice Across Age and Strain

The NASA GeneLab Sample Processing Laboratory offers high-throughput sequencing services to NASA-funded space biology researchers. Space biology studies have specific challenges such as low sample numbers, introducing susceptibility to batch effects from sample handling. These issues are compounded by complex protocols such as single-nuclei isolation and sequencing, which has recently become an attractive methodology for assessing the cellular diversity within spaceflight samples. High quality single-nuclei sequencing requires reproducible protocols to dissociate tissue and generate clean suspension of intact single nuclei. Producing single-nuclei suspension from brain tissue is particularly challenging due to cell type heterogeneity and the myelin sheath that carries over into the nuclei suspension as debris. Current procedures tend to be time consuming and sometimes include steps that can alter gene expression and create cell-type bias. Commercially available nuclei isolation kits, such as the 10X Genomics nuclei isolation kit, offers a streamlined way to process samples for nuclei isolation, thereby minimizing batch effects and enabling reproducibility. In this study, we report on the performance of the 10X Genomics nuclei isolation kit and Chromium Next GEM Single Cell Multiome ATAC + Gene Expression kit to generate sequencing libraries from space-flown mouse brain samples. Single nuclei sequencing was performed on frozen mouse brain tissue from two spaceflight missions, Rodent Research-10 (RR-10) and RR Reference Mission-2 (RRRM-2). RR-10 mice were female B6129SF2/J, euthanized at 18-19 weeks whereas RRRM-2 mice were female C57BL/6NTac, euthanized at 20 or 37 weeks. Sequencing data was processed using standard GeneLab data processing pipelines. We report evaluation of the performance of the 10X Genomics nuclei isolation kit for spaceflight samples from mouse brain, and evaluation of reproducibility across different mouse strains and age groups. We also report preliminary scientific results including cell type inference, cell clustering, and differentially expressed genes and pathways between spaceflight and ground control samples.

RR-10↗