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At least 973 records · Page 54

Integration of pH Control into Chi.Bio Reactors and Demonstration with Small-Scale Enzymatic Poly(ethylene terephthalate) Hydrolysis

Small-scale bioreactors that are affordable and accessible would be of major benefit to the research community. In previous work, an open-source, automated bioreactor system was designed to operate up to the 30 mL scale with online optical monitoring, stirring, and temperature control, and this system, dubbed Chi.Bio, is now commercially available at a cost that is typically 1–2 orders of magnitude less than commercial bioreactors. In this work, we further expand the capabilities of the Chi.Bio system by enabling continuous pH monitoring and control through hardware and software modifications. For hardware modifications, we sourced low-cost, commercial pH circuits and made straightforward modifications to the Chi.Bio head plate to enable continuous pH monitoring. For software integration, we introduced closed-loop feedback control of the pH measured inside the Chi.Bio reactors and integrated a pH-control module into the existing Chi.Bio user interface. We demonstrated the utility of pH control through the small-scale depolymerization of the synthetic polyester, poly(ethylene terephthalate) (PET), using a benchmark cutinase enzyme, and compared this to 250 mL bioreactor hydrolysis reactions. The results in terms of PET conversion and rate, measured both by base addition and product release profiles, are statistically equivalent, with the Chi.Bio system allowing for a 20-fold reduction of purified enzyme required relative to the 250 mL bioreactor setup. Through inexpensive modifications, the ability to conduct pH control in Chi.Bio reactors widens the potential slate of biochemical reactions and biological cultivations for study in this system, and may also be adapted for use in other bioreactor platforms.

09 BIOMASS FUELS↗

NASA's Rodent Research Project: Validation of Capabilities for Conducting Long Duration Experiments in Space

Research using rodents is an essential tool for advancing biomedical research on Earth and in space. Prior rodent experiments on the Shuttle were limited by the short flight duration. The International Space Station (ISS) provides a new platform for conducting rodent experiments under long duration conditions. Rodent Research (RR)-1 was conducted to validate flight hardware, operations, and science capabilities that were developed at the NASA Ames Research Center. Twenty C57BL6J adult female mice were launched on Sept 21, 2014 in a Dragon Capsule (SpaceX-4), then transferred to the ISS for a total time of 21-22 days (10 commercial mice) or 37 days (10 validation mice). Tissues collected on-orbit were either rapidly frozen or preserved in RNAlater at -80C (n2group) until their return to Earth. Remaining carcasses on-orbit were rapidly frozen for dissection post-flight. The three controls groups at Kennedy Space Center consisted of: Basal mice euthanized at the time of launch, Vivarium controls housed in standard cages, and Ground Controls (GC) housed in flight hardware within an environmental chamber. Upon return to Earth, there were no differences in body weights between Flight (FLT) and GC at the end of the 37 days in space. Liver enzyme activity levels of FLT mice and all control mice were similar in magnitude to those of the samples that were processed under optimal conditions in the laboratory. Liver samples dissected on-orbit yielded high quality RNA (RIN8.99+-0.59, n7). Liver samples dissected post-flight from the intact, frozen FLT carcasses yielded RIN of 7.27 +- 0.52 (n6). Additionally, wet weights of various tissues were measured. Adrenal glands and spleen showed no significant differences in FLT compared to GC although thymus and livers weights were significantly greater in FLT compared to GC. Over 3,000 tissue aliquots collected post-flight from the four groups of mice were deposited into the Ames Life Science Data Archives for future Biospecimen Sharing Program. Together, the RR validation flight successfully demonstrates the capability to support long-duration experimentation on the ISS to achieve both basic science and biomedical objectives.

spaceflight↗

Microbial diversity analysis of two full-scale seawater desalination treatment trains provides insights into detrimental biofilm formation

Detrimental biofilms on RO membranes remain a crucial challenge for seawater desalination. Comparative analysis of 16S rRNA gene amplicon sequencing data revealed differences and commonalities of biofilm communities associated with unit operations in the two largest seawater desalination facilities in the U.S., the Claude "Bud" Lewis Carlsbad Desalination Plant and the Tampa Bay Seater Desalination facility. At both plants, feedwater collected at a single time point was a poor indicator of the RO membrane communities, which showed far greater taxa diversity. The analysis of prefilter cartridges from the Carlsbad plant revealed similarly high taxon diversity as the RO module biofilms, with relevant differences. Algal sequences were enriched on the prefilter cartridges as were sequences representing Bdellovibrionota, which are predatory bacteria. Sequences representing opportunistic Gammaproteobacteria (i.e., Shewanella, Woesia) were present in significantly higher relative abundance on the RO membranes than in the prefilter cartridges, suggesting growth of certain taxa in the RO modules. Untargeted metabolomics distinguished intra- and inter-desalination plant biofilm samples, highlighting the potential value of this tool for biofilm monitoring. These findings underscore the value of omics tools for effective microbial monitoring, to understand biofouling dynamics within RO desalination plants, and to provide insight for the development of ecologically-informed biofilm control measures.

59 BASIC BIOLOGICAL SCIENCES↗

Evaluation of sterol-o-acyl transferase 1 and cholesterol ester levels in plasma, peritoneal fluid and tumor tissue of patients with endometrial cancer: A pilot study

Endometrial cancer (EC) is the most prevalent gynecological malignancy. Abnormal accumulation of sterol-O-acyl transferase 1 (SOAT1) and SOAT1-mediated cholesterol ester (CE) contributes to cancer progression in various malignancies, including ovarian cancer. Therefore, it was hypothesized that similar molecular changes may occur in EC. The present study aimed to evaluate the diagnostic and/or prognostic potential of SOAT1 and CE in EC by: i) Determining SOAT1 and CE levels in plasma, peritoneal fluid and endometrial tissue from patients with EC and control subjects; ii) performing receiver operating characteristic curve analysis to determine diagnostic performance; iii) comparing SOAT1 and CE expression to that of the tumor proliferation marker Ki67; and iv) assessing the association between SOAT1 expression and survival. Enzyme-linked immunosorbent assay was used to determine the levels of SOAT1 protein in tissue, plasma and peritoneal fluid. The mRNA and protein expression levels of SOAT1 and Ki67 in tissues were detected by reverse transcription-quantitative polymerase chain reaction and immunohistochemistry, respectively. CE levels were determined colorimetrically in plasma and peritoneal fluid. SOAT1-associated survival data from the cBioPortal cancer genomics database were used to assess prognostic relevance. The results revealed that SOAT1 and CE levels were significantly elevated in tumor tissue and peritoneal fluid samples collected from the EC group. By contrast, the plasma levels of SOAT1 and CE in the EC and control groups were similar. Significant positive associations between CE and SOAT1, SOAT1/CE and Ki67, and SOAT1/CE and poor overall survival in patients with EC suggested that SOAT1/CE may be associated with malignancy, aggressiveness and poor prognosis. In conclusion, SOAT1 and CE may serve as potential biomarkers for prognosis and target-specific treatment of EC.

59 BASIC BIOLOGICAL SCIENCES↗

Detection of Head and Neck Cancer Based on Longitudinal Changes in Serum Protein Abundance

Head and Neck Squamous Cell Carcinoma (HNSCC) is significantly represented in the military population with substantial impact on military operations. Two-thirds of new HNSCC cases are first diagnosed at advanced stage III or IV with lymph node metastases. Thus, there is an urgent need of biomarkers for early detection and risk stratification. Archived serum samples from the Department of Defense Serum Repository provide a unique resource for longitudinal studies of HNSCC in the military and the identification of early detection biomarkers. Methods: We identified 175 patients diagnosed with HNSCC with 175 matched healthy controls and retrieved a total of 978 serum samples drawn at the time of diagnosis, 2 and 4 years prior to diagnosis, and 2 years after diagnosis. Following immunoaffinity depletion, serum samples were analyzed by targeted proteomics assays for multiplexed quantification of a panel of 146 candidate protein biomarkers from the curated literature. Results: We derived a 13-protein signature that distinguishes case versus controls based on longitudinal changes in serum protein concentration. The abundances of each of the 13 proteins remain constant over time in control subjects. The area under the curve for the derived random forest classifier was 0.90. Conclusions: This 13-protein classifier is highly promising for detection of HNSCC prior to overt symptoms. Impact: Use of longitudinal samples has significant potential to identify biomarkers for detection and risk stratification.

59 BASIC BIOLOGICAL SCIENCES↗

Foldable Instrumented Bits for Ultrasonic/Sonic Penetrators

Long tool bits are undergoing development that can be stowed compactly until used as rock- or ground-penetrating probes actuated by ultrasonic/sonic mechanisms. These bits are designed to be folded or rolled into compact form for transport to exploration sites, where they are to be connected to their ultrasonic/ sonic actuation mechanisms and unfolded or unrolled to their full lengths for penetrating ground or rock to relatively large depths. These bits can be designed to acquire rock or soil samples and/or to be equipped with sensors for measuring properties of rock or soil in situ. These bits can also be designed to be withdrawn from the ground, restowed, and transported for reuse at different exploration sites. Apparatuses based on the concept of a probe actuated by an ultrasonic/sonic mechanism have been described in numerous prior NASA Tech Briefs articles, the most recent and relevant being "Ultrasonic/ Sonic Impacting Penetrators" (NPO-41666) NASA Tech Briefs, Vol. 32, No. 4 (April 2008), page 58. All of those apparatuses are variations on the basic theme of the earliest ones, denoted ultrasonic/sonic drill corers (USDCs). To recapitulate: An apparatus of this type includes a lightweight, low-power, piezoelectrically driven actuator in which ultrasonic and sonic vibrations are generated and coupled to a tool bit. The combination of ultrasonic and sonic vibrations gives rise to a hammering action (and a resulting chiseling action at the tip of the tool bit) that is more effective for drilling than is the microhammering action of ultrasonic vibrations alone. The hammering and chiseling actions are so effective that the size of the axial force needed to make the tool bit advance into soil, rock, or another material of interest is much smaller than in ordinary twist drilling, ordinary hammering, or ordinary steady pushing. Examples of properties that could be measured by use of an instrumented tool bit include electrical conductivity, permittivity, magnetic field, magnetic permeability, temperature, and any other properties that can be measured by fiber-optic sensors. The problem of instrumenting a probe of this type is simplified, relative to the problem of attaching electrodes in a rotating drill bit, in two ways: (1) Unlike a rotating drill bit, a bit of this type does not have flutes, which would compound the problem of ensuring contact between sensors and the side wall of a hole; and (2) there is no need for slip rings for electrical contact between sensor electronic circuitry and external circuitry because, unlike a rotating drill, a tool bit of this type is not rotated continuously during operation. One design for a tool bit of the present type is a segmented bit with a segmented, hinged support structure (see figure). The bit and its ultrasonic/sonic actuator are supported by a slider/guiding fixture, and its displacement and preload are controlled by a motor. For deployment from the folded configuration, a spring-loaded mechanism rotates the lower segment about the hinges, causing the lower segment to become axially aligned with the upper segment. A latching mechanism then locks the segments of the bit and the corresponding segments of the slider/guiding fixture. Then the entire resulting assembly is maneuvered into position for drilling into the ground. Another design provides for a bit comprising multiple tubular segments with an inner alignment string, similar to a foldable tent pole comprising multiple tubular segments with an inner elastic cable connecting the two ends. At the beginning of deployment, all segments except the first (lowermost) one remain folded, and the ultrasonic/sonic actuator is clamped to the top of the lowermost segment and used to drive this segment into the ground. When the first segment has penetrated to a specified depth, the second segment is connected to the upper end of the first segment to form a longer rigid tubular bit and the actuator is moved to the upper end of the second segnt. The process as described thus far is repeated, adding segments until the desired depth of penetration has been attained. Yet other designs provide for bits in the form of bistable circular- or rectangular- cross-section tubes that can be stowed compactly like rolls of flat tape and become rigidified upon extension to full length, in a manner partly similar to that of a common steel tape measure. Albeit not marketed for use in tool bits, a bistable reeled composite product that transforms itself from a flat coil to a rigid tube of circular cross section when unrolled, is commercially available under the trade name RolaTube(TradeMark) and serves as a model for the further development of tool bits of this subtype.

Bar-Cohen, Yoseph↗

Two-Armed, Mobile, Sensate Research Robot

The Anthropomorphic Robotic Testbed (ART) is an experimental prototype of a partly anthropomorphic, humanoid-size, mobile robot. The basic ART design concept provides for a combination of two-armed coordination, tactility, stereoscopic vision, mobility with navigation and avoidance of obstacles, and natural-language communication, so that the ART could emulate humans in many activities. The ART could be developed into a variety of highly capable robotic assistants for general or specific applications. There is especially great potential for the development of ART-based robots as substitutes for live-in health-care aides for home-bound persons who are aged, infirm, or physically handicapped; these robots could greatly reduce the cost of home health care and extend the term of independent living. The ART is a fully autonomous and untethered system. It includes a mobile base on which is mounted an extensible torso topped by a head, shoulders, and two arms. All subsystems of the ART are powered by a rechargeable, removable battery pack. The mobile base is a differentially- driven, nonholonomic vehicle capable of a speed >1 m/s and can handle a payload >100 kg. The base can be controlled manually, in forward/backward and/or simultaneous rotational motion, by use of a joystick. Alternatively, the motion of the base can be controlled autonomously by an onboard navigational computer. By retraction or extension of the torso, the head height of the ART can be adjusted from 5 ft (1.5 m) to 6 1/2 ft (2 m), so that the arms can reach either the floor or high shelves, or some ceilings. The arms are symmetrical. Each arm (including the wrist) has a total of six rotary axes like those of the human shoulder, elbow, and wrist joints. The arms are actuated by electric motors in combination with brakes and gas-spring assists on the shoulder and elbow joints. The arms are operated under closed-loop digital control. A receptacle for an end effector is mounted on the tip of the wrist and contains a force-and-torque sensor that provides feedback for force (compliance) control of the arm. The end effector could be a tool or a robot hand, depending on the application.

Engelberger, J. F.↗

The military gear microbiome: risk factors surrounding the warfighter

ABSTRACT Combat extremity wounds are highly susceptible to contamination from surrounding environmental material. This bioburden could be partially transferred from materials in immediate proximity to the wound, including fragments of the uniform and gear. However, the assessment of the microbial bioburden present on military gear during operational conditions of deployment or training is relatively unexplored. Opportunistic pathogens that can survive on gear represent risk factors for infection following injury, especially following combat blasts, where fibers and other materials are embedded in wounded tissue. We utilized 16S rRNA sequencing to assess the microbiome composition of different military gear types (boot, trouser, coat, and canteen) from two operational environments (training in Hawai’i and deployed in Indonesia) across time (days 0 and 14). We found that microbiome diversity, stability, and composition were dependent on gear type, training location, and sampling timepoint. At day 14, species diversity was significantly higher in Hawai’i samples compared to Indonesia samples for boot, coat, and trouser swabs. In addition, we observed the presence of potential microbial risk factors, as opportunistic pathogenic species, such as Acinetobacter , Pseudomonas, and Staphylococcus , were found to be present in all sample types and in both study sites. These study outcomes will be used to guide the design of antimicrobial materials and uniforms and for infection control efforts following combat blasts and other injuries, thereby improving treatment guidance during military training and deployment. IMPORTANCE Combat extremity wounds are vulnerable to contamination from environments of proximity to the warfighter, leading to potential detrimental outcomes such as infection and delayed wound healing. Therefore, microbial surveillance of such environments is necessary to aid the advancement of military safety and preparedness through clinical diagnostics, treatment protocols, and uniform material design.

59 BASIC BIOLOGICAL SCIENCES↗

The Design and Implementation of a Secure Datastore Based on Ethereum Smart Contract

In this paper, we present a secure datastore based on an Ethereum smart contract. Our research is guided by three research questions. First, we will explore to what extend a smart-contract-based datastore should resemble a traditional database system. Second, we will investigate how to store the data in a smart-contract-based datastore for maximum flexibility while minimizing the gas consumption. Third, we seek answers regarding whether or not a smart-contract-based datastore should incorporate complex processing such as data encryption and data analytic algorithms. The proposed smart-contract-based datastore aims to strike a good balance between several constraints: (1) smart contracts are publicly visible, which may create a confidentiality concern for the data stored in the datastore; (2) unlike traditional database systems, the Ethereum smart contract programming language (i.e., Solidity) offers very limited data structures for data management; (3) all operations that mutate the blockchain state would incur financial costs and the developers for smart contracts must make sure sufficient gas is provisioned for every smart contract call, and ideally, the gas consumption should be minimized. Our investigation shows that although it is essential for a smart-contract-based datastore to offer some basic data query functionality, it is impractical to offer query flexibility that resembles that of a traditional database system. Furthermore, we propose that data should be structured as tag-value pairs, where the tag serves as a non-unique key that describes the nature of the value. We also conclude that complex processing should not be allowed in the smart contract due to the financial burden and security concerns. The tag-based secure datastore designed this way also defines its applicative perimeter, i.e., only applications that align with our strategy would find the proposed datastore a good fit. Those that would rather incur higher financial cost for more data query flexibility and/or less user burden on data pre- and post-processing would find the proposed database too restrictive.

32 ENERGY CONSERVATION, CONSUMPTION, AND UTILIZATI↗

Viral infections likely mediate microbial controls on ecosystem responses to global warming

Abstract Climate change is affecting how energy and matter flow through ecosystems, thereby altering global carbon and nutrient cycles. Microorganisms play a fundamental role in carbon and nutrient cycling and are thus an integral link between ecosystems and climate. Here, we highlight a major black box hindering our ability to anticipate ecosystem climate responses: viral infections within complex microbial food webs. We show how understanding and predicting ecosystem responses to warming could be challenging—if not impossible—without accounting for the direct and indirect effects of viral infections on different microbes (bacteria, archaea, fungi, protists) that together perform diverse ecosystem functions. Importantly, understanding how rising temperatures associated with climate change influence viruses and virus-host dynamics is crucial to this task, yet is severely understudied. In this perspective, we (i) synthesize existing knowledge about virus-microbe-temperature interactions and (ii) identify important gaps to guide future investigations regarding how climate change might alter microbial food web effects on ecosystem functioning. To provide real-world context, we consider how these processes may operate in peatlands—globally significant carbon sinks that are threatened by climate change. We stress that understanding how warming affects biogeochemical cycles in any ecosystem hinges on disentangling complex interactions and temperature responses within microbial food webs.

54 ENVIRONMENTAL SCIENCES↗

Unlocking the magic in mycelium: Using synthetic biology to optimize filamentous fungi for biomanufacturing and sustainability

Filamentous fungi drive carbon and nutrient cycling across our global ecosystems, through its interactions with growing and decaying flora and their constituent microbiomes. The remarkable metabolic diversity, secretion ability, and fiber-like mycelial structure that have evolved in filamentous fungi have been increasingly exploited in commercial operations. The industrial potential of mycelial fermentation ranges from the discovery and bioproduction of enzymes and bioactive compounds, the decarbonization of food and material production, to environmental remediation and enhanced agricultural production. Despite its fundamental impact in ecology and biotechnology, molds and mushrooms have not, to-date, significantly intersected with synthetic biology in ways comparable to other industrial cell factories (e.g. Escherichia coli,Saccharomyces cerevisiae, and Komagataella phaffii). In this review, we summarize a suite of synthetic biology and computational tools for the mining, engineering and optimization of filamentous fungi as a bioproduction chassis. A combination of methods across genetic engineering, mutagenesis, experimental evolution, and computational modeling can be used to address strain development bottlenecks in established and emerging industries. These include slow mycelium growth rate, low production yields, non-optimal growth in alternative feedstocks, and difficulties in downstream purification. In the scope of biomanufacturing, we then detail previous efforts in improving key bottlenecks by targeting protein processing and secretion pathways, hyphae morphogenesis, and transcriptional control. Bringing synthetic biology practices into the hidden world of molds and mushrooms will serve to expand the limited panel of host organisms that allow for commercially-feasible and environmentally-sustainable bioproduction of enzymes, chemicals, therapeutics, foods, and materials of the future.

59 BASIC BIOLOGICAL SCIENCES↗

Space Station technology testbed: 2010 deep space transport

A space station in a crew-tended or permanently crewed configuration will provide major R&D opportunities for innovative, technology and materials development and advanced space systems testing. A space station should be designed with the basic infrastructure elements required to grow into a major systems technology testbed. This space-based technology testbed can and should be used to support the development of technologies required to expand our utilization of near-Earth space, the Moon and the Earth-to-Jupiter region of the Solar System. Space station support of advanced technology and materials development will result in new techniques for high priority scientific research and the knowledge and R&D base needed for the development of major, new commercial product thrusts. To illustrate the technology testbed potential of a space station and to point the way to a bold, innovative approach to advanced space systems' development, a hypothetical deep space transport development and test plan is described. Key deep space transport R&D activities are described would lead to the readiness certification of an advanced, reusable interplanetary transport capable of supporting eight crewmembers or more. With the support of a focused and highly motivated, multi-agency ground R&D program, a deep space transport of this type could be assembled and tested by 2010. Key R&D activities on a space station would include: (1) experimental research investigating the microgravity assisted, restructuring of micro-engineered, materials (to develop and verify the in-space and in-situ 'tuning' of materials for use in debris and radiation shielding and other protective systems), (2) exposure of microengineered materials to the space environment for passive and operational performance tests (to develop in-situ maintenance and repair techniques and to support the development, enhancement, and implementation of protective systems, data and bio-processing systems, and virtual reality and telepresence/kinetic processes), (3) subsystem tests of advanced nuclear power, nuclear propulsion and communication systems (using boom extensions, remote station-keeping platforms and mobile EVA crew and robots), and (4) logistics support (crew and equipment) and command and control of deep space transport assembly, maintenance, and refueling (using a station-keeping platform).

Holt, Alan C.↗

The X-43A Flight Research Program: Lessons Learned on the Road to Mach 10

During an aerospace engineer's undergraduate studies, he or she will attend classes in aerodynamics, thermodynamics, structures, stability and control, dynamics, design, propulsion, and computer science, along with the related courses in mathematics, physics, statistics, and chemistry required to understand the material. Upon graduation, the new engineer will have acquired a basic knowledge of how to build an aerospace vehicle. What only comes through experience, however, is the understanding of the inevitable imperfect process through which an aerospace vehicle is built. This is the adventure of turning a basic concept into functional hardware. Engineers working on a project must often deal with ambiguous situations. They are routinely asked by management to provide risk assessments of a project, yet even after careful analysis uncertainties remain. The project must be accomplished within finite limits of time and money. The question an engineer answers is whether the solution to potential problem is worth the cost and schedule delay, or if the solution might actually be worse than the problem it is meant to solve. Review protocols are established to ensure that an unknown has not been overlooked. But these cannot protect against an unknown unknown. Examples of these situations can be found in the history of the X-43A Hyper-X (Hypersonic Experiment) program. In this NASA project, a supersonic combustion ramjet (scramjet) engine was flight tested on a subscale vehicle. The X-43A Hyper-X Research Vehicle (HXRV) was launched from a B-52B mothership, then boosted to the test speed by a modified Pegasus rocket first stage, called the Hyper-X Launch Vehicle (HXLV). Once at the proper speed and altitude, the X-43A separated from the booster, stabilized itself, and then the engine test began. Although wind-tunnel scramjet engine tests had begun in the late 1950s, before the Hyper-X program there had never been an actual in-flight test of such an engine integrated with an appropriate airframe. Thus, while the scramjet had successfully operated in the artificial airflow of wind tunnels, the concept had yet to be proven in real air. These conditions meant changes in density and temperature, as well as changes in angle of attack and sideslip of a free-flying vehicle. A wind tunnel is limited in its ability to simulate these subtle factures, which have a major impact on almost any vehicle, but especially that of a scramjet's performance. The Hyper-X project was to provide a real-world benchmark of the ground test data. The full scale X-43A engine would be operated in the wind tunnel, and then flown, and the data from its operation would then be compared with projections. If these matched, the wind tunnel data would be considered a reliable design tool for future scramjet. If there were significant differences, the reasons for these would have to be identified. Until such information was available, scramjets would lack the technological maturity to be considered for future space launch or high-speed atmospheric flight vehicles.

Peebles, Curtis↗

NASA's Rodent Research Project: Validation of Flight Hardware, Operations and Science Capabilities for Conducting Long Duration Experiments in Space

Research using rodents is an essential tool for advancing biomedical research on Earth and in space. Rodent Research (RR)-1 was conducted to validate flight hardware, operations, and science capabilities that were developed at the NASA Ames Research Center. Twenty C57BL/6J adult female mice were launched on Sept 21, 2014 in a Dragon Capsule (SpaceX-4), then transferred to the ISS for a total time of 21-22 days (10 commercial mice) or 37 (10 validation mice). Tissues collected on-orbit were either rapidly frozen or preserved in RNA later at less than or equal to -80 C (n=2/group) until their return to Earth. Remaining carcasses were rapidly frozen for dissection post-flight. The three controls groups at Kennedy Space Center consisted of: Basal mice euthanized at the time of launch, Vivarium controls, housed in standard cages, and Ground Controls (GC), housed in flight hardware within an environmental chamber. FLT mice appeared more physically active on-orbit than GC, and behavior analysis are in progress. Upon return to Earth, there were no differences in body weights between FLT and GC at the end of the 37 days in space. RNA was of high quality (RIN greater than 8.5). Liver enzyme activity levels of FLT mice and all control mice were similar in magnitude to those of the samples that were optimally processed in the laboratory. Liver samples collected from the intact frozen FLT carcasses had RNA RIN of 7.27 +/- 0.52, which was lower than that of the samples processed on-orbit, but similar to those obtained from the control group intact carcasses. Nonetheless, the RNA samples from the intact carcasses were acceptable for the most demanding transcriptomic analyses. Adrenal glands, thymus and spleen (organs associated with stress response) showed no significant difference in weights between FLT and GC. Enzymatic activity was also not significantly different. Over 3,000 tissues collected from the four groups of mice have become available for the Biospecimen Sharing Program. Together, these validation flight findings demonstrate the capability to support long-duration RR on the ISS to achieve both basic science and biomedical objectives.

rodents↗

Evaluation of Plasma Phosphorylated Tau217 for Differentiation Between Alzheimer Disease and Frontotemporal Lobar Degeneration Subtypes Among Patients With Corticobasal Syndrome

Plasma phosphorylated tau217 (p-tau217), a biomarker of Alzheimer disease (AD), is of special interest in corticobasal syndrome (CBS) because autopsy studies have revealed AD is the driving neuropathology in up to 40% of cases. This differentiates CBS from other 4-repeat tauopathy (4RT)–associated syndromes, such as progressive supranuclear palsy Richardson syndrome (PSP-RS) and nonfluent primary progressive aphasia (nfvPPA), where underlying frontotemporal lobar degeneration (FTLD) is typically the primary neuropathology. To validate plasma p-tau217 against positron emission tomography (PET) in 4RT-associated syndromes, especially CBS. This multicohort study with 6, 12, and 24-month follow-up recruited adult participants between January 2011 and September 2020 from 8 tertiary care centers in the 4RT Neuroimaging Initiative (4RTNI). All participants with CBS (n = 113), PSP-RS (n = 121), and nfvPPA (n = 39) were included; other diagnoses were excluded due to rarity (n = 29). Individuals with PET-confirmed AD (n = 54) and PET-negative cognitively normal control individuals (n = 59) were evaluated at University of California San Francisco. Operators were blinded to the cohort. Plasma p-tau217, measured by Meso Scale Discovery electrochemiluminescence, was validated against amyloid-β (Aβ) and flortaucipir (FTP) PET. Imaging analyses used voxel-based morphometry and bayesian linear mixed-effects modeling. Clinical biomarker associations were evaluated using longitudinal mixed-effect modeling. Of 386 participants, 199 (52%) were female, and the mean (SD) age was 68 (8) years. Plasma p-tau217 was elevated in patients with CBS with positive Aβ PET results (mean [SD], 0.57 [0.43] pg/mL) or FTP PET (mean [SD], 0.75 [0.30] pg/mL) to concentrations comparable to control individuals with AD (mean [SD], 0.72 [0.37]), whereas PSP-RS and nfvPPA showed no increase relative to control. Within CBS, p-tau217 had excellent diagnostic performance with area under the receiver operating characteristic curve (AUC) for Aβ PET of 0.87 (95% CI, 0.76-0.98; P < .001) and FTP PET of 0.93 (95% CI, 0.83-1.00; P < .001). At baseline, individuals with CBS-AD (n = 12), defined by a PET-validated plasma p-tau217 cutoff 0.25 pg/mL or greater, had increased temporoparietal atrophy at baseline compared to individuals with CBS-FTLD (n = 39), whereas longitudinally, individuals with CBS-FTLD had faster brainstem atrophy rates. Individuals with CBS-FTLD also progressed more rapidly on a modified version of the PSP Rating Scale than those with CBS-AD (mean [SD], 3.5 [0.5] vs 0.8 [0.8] points/year; P = .005). In this cohort study, plasma p-tau217 had excellent diagnostic performance for identifying Aβ or FTP PET positivity within CBS with likely underlying AD pathology. Plasma P-tau217 may be a useful and inexpensive biomarker to select patients for CBS clinical trials.

59 BASIC BIOLOGICAL SCIENCES↗

Improved Blackbody Temperature Sensors for a Vacuum Furnace

Some improvements have been made in the design and fabrication of blackbody sensors (BBSs) used to measure the temperature of a heater core in a vacuum furnace. Each BBS consists of a ring of thermally conductive, high-melting-temperature material with two tantalum-sheathed thermocouples attached at diametrically opposite points. The name "blackbody sensor" reflects the basic principle of operation. Heat is transferred between the ring and the furnace heater core primarily by blackbody radiation, heat is conducted through the ring to the thermocouples, and the temperature of the ring (and, hence, the temperature of the heater core) is measured by use of the thermocouples. Two main requirements have guided the development of these BBSs: (1) The rings should have as high an emissivity as possible in order to maximize the heat-transfer rate and thereby maximize temperature-monitoring performance and (2) the thermocouples must be joined to the rings in such a way as to ensure long-term, reliable intimate thermal contact. The problem of fabricating a BBS to satisfy these requirements is complicated by an application-specific prohibition against overheating and thereby damaging nearby instrumentation leads through the use of conventional furnace brazing or any other technique that involves heating the entire BBS and its surroundings. The problem is further complicated by another application-specific prohibition against damaging the thin tantalum thermocouple sheaths through the use of conventional welding to join the thermocouples to the ring. The first BBS rings were made of graphite. The tantalum-sheathed thermocouples were attached to the graphite rings by use of high-temperature graphite cements. The ring/thermocouple bonds thus formed were found to be weak and unreliable, and so graphite rings and graphite cements were abandoned. Now, each BBS ring is made from one of two materials: either tantalum or a molybdenum/titanium/zirconium alloy. The tantalum-sheathed thermocouples are bonded to the ring by laser brazing. The primary advantage of laser brazing over furnace brazing is that in laser brazing, it is possible to form a brazed connection locally, without heating nearby parts to the flow temperature of the brazing material. Hence, it is possible to comply with the prohibition against overheating nearby instrumentation leads. Also, in laser brazing, unlike in furnace brazing, it is possible to exert control over the thermal energy to such a high degree that it becomes possible to braze the thermocouples to the ring without burning through the thin tantalum sheaths on the thermocouples. The brazing material used in the laser brazing process is a titanium-boron paste. This brazing material can withstand use at temperatures up to about 1,400 C. In thermal-cycling tests performed thus far, no debonding between the rings and thermocouples has been observed. Emissivity coatings about 0.001 in. (.0.025 mm) thick applied to the interior surfaces of the rings have been found to improve the performance of the BBS sensors by raising the apparent emissivities of the rings. In thermal-cycling tests, the coatings were found to adhere well to the rings.

Farmer, Jeff↗

Evaluating Movement Patterns of the Rattlesnake Hills Elk Herd on Hanford for Calendar Years 2019-2024

Biologists first documented elk on the Hanford Site in 1972; since then, the elk herd known as the Rattlesnake Hills Elk Herd (RHEH) has grown substantially (PNNL-13331, Population Characteristics and Seasonal Movement Patterns of the Rattlesnake Hills Elk Herd: Status Report 2000). Through the 1990s, the core range of the RHEH was focused on the portion of the Hanford Reach National Monument known as the Fitzner-Eberhardt Arid Lands Ecology (ALE) Reserve. More recently, larger numbers of elk have been occupying the U.S. Department of Energy (DOE), Hanford Field Office (HFO), formerly the DOE, Richland Operations Office managed portion- of the Hanford Site, known as central Hanford (Figure 1-1). This began with bachelor groups of bulls occupying the site intermittently, and the herd has grown to resident herds, including bulls, cows, and calves. The Washington Department of Fish and Wildlife (WDFW) established a target herd size for the RHEH of less than 350 animals to minimize damages on adjacent private agricultural lands (Washington State Elk Herd Plan–Yakima Elk Herd [WDFW 2002]). Attempts to control the RHEH population through hunting on private lands and a relocation effort during 2000 have failed to limit growth of the population toward the WDFW target, and the herd exceeds 1,600 according to recent counts (PNNL-13331; DOE/RL- 2023-20, Hanford Annual Site Environmental Report for Calendar Year 2022). Although elk are present on central Hanford year-round, they continue to move between areas offsite, the ALE Reserve, and central Hanford throughout the year, and it is these movements that result in many of the elk-vehicle collisions (EVC) that occur along Hanford Site roads and on the Washington State highways bordering the site. In addition, changes in management strategies on the ALE Reserve, including recent and potential future tribal elk hunts, may alter herd behavior and result in additional animals transiting onto and off central Hanford.

54 ENVIRONMENTAL SCIENCES↗

A Joint Experimental/Computational Study of Non-Idealities in Practical Rotating Detonation Engines. Final Project Report, 10/2015 - 09/2020

The overall goal of this work is to understand non-ideal effects in practical rotating detonation engines (RDEs) that impede the realization of theoretical detonation cycle efficiencies. The main limitations in RDE operability and performance are associated with practical design considerations, where non-idealities, such as incomplete mixing, fuel leakage, secondary deflagration, instabilities and geometry-dependent effects, reduce the effectiveness of the detonation-based compression cycle. The goal of this work is to take a fundamental point of view, and use detailed experiments and simulations to understand non-ideal effects, their contribution to loss in pressure gain, and RDE operability limitations. Current research is mainly focused on design and overall operability of RDEs. Our approach is instead to look at the physics of basic processes occurring in RDEs. In particular, we consider processes associated with non-idealities and we use a combination of laser and non-laser diagnostics and large eddy simulation/direct numerical simulation computations to investigate these underlying phenomena. By advancing the fundamental science of detonation wave propagation in RDE flowfields, and subject to non-ideal conditions, will enable design to progress more rapidly. This work is a collaborative work involving experiments, simulations and external domain experts to understand the physics behind RDEs and non-idealities. Through collaboration between experimentalists and modelers, we have developed targeted experiments to investigate fundamental aspects of the operation of an RDE. Furthermore, we have developed advanced combustion models that account for non-ideal effects in the operation of an RDE and are able to capture the complexities of realistic RDE configurations. We conduct detailed physical and computational experiments to investigate the flame structure ahead and behind detonation fronts in the presence of non-uniform or incomplete mixing, leakage and secondary combustion. We use optical and laser diagnostics to identify the structure of the detonation wave. We consider different working parameters, such injection scheme, geometry, flow rates and equivalence ratios. We brought in DNS/LES capabilities to RDE design by specifically incorporating detailed kinetics and pressure-sensitive combustion physics, thus improving upon current practice based on an Euler equations description that do not take into account viscous effects, while the combustion process is introduced using an induction time based global ignition model. These current models lack the capability of capturing non-ideal mixing and fuel stratification effects — limitations that models and analysis developed in this work have been reduced. Furthermore, the significant changes in pressure within the domain due to the detonation requires combustion models with time-varying pressure effects. In this regard, using DNS with detailed chemistry, and developing LES-based combustion models for complex geometries with detonation wave has enable to understand some of the underlying phenomena controlling the operation of these devices. The main objectives developed by the project are the following ones: Objective 1. Develop canonical and operational RDE configurations, as well as imaging-based diagnostics for understanding fuel stratification, leakage, deflagration and detonation structure under non-ideal conditions in RDEs. Objective 2. Develop detailed computational tools for studying detonation wave propagation processes in RDEs. Objective 3. Develop a comprehensive picture of the fundamental physics governing non-idealities and how they impact RDE performance and operability from both experiments and simulations.

03 NATURAL GAS↗