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All sourcesNASA NTRSDOE OSTI
NASA NTRS2016

Successful Validation of Sample Processing and Quantitative Real-Time PCR Capabilities on the International Space Station

The WetLab-2 system was developed by NASA Ames Research Center to offer new capabilities to researchers. The system can lyse cells and extract RNA (Ribonucleic Acid) on-orbit from different sample types ranging from microbial cultures to animal tissues. The purified RNA can then either be stabilized for return to Earth or can be used to conduct on-orbit quantitative Reverse Transcriptase PCR (Polymerase Chain Reaction) (qRT-PCR) analysis without the need for sample return. The qRT-PCR results can be downlinked to the ground a few hours after the completion of the run. The validation flight of the WetLab-2 system launched on SpaceX-8 on April 8, 2016. On orbit operations started on April 15th with system setup and was followed by three quantitative PCR runs using an E. coli genomic DNA template pre-loaded at three different concentrations. These runs were designed to discern if quantitative PCR functions correctly in microgravity and if the data is comparable to that from the ground control runs. The flight data showed no significant differences compared to the ground data though there was more variability in the values, this was likely due to the numerous small bubbles observed. The capability of the system to process samples and purify RNA was then validated using frozen samples prepared on the ground. The flight data for both E. coli and mouse liver clearly shows that RNA was successfully purified by our system. The E. coli qRT-PCR run showed successful singleplex, duplex and triplex capability. Data showed high variability in the resulting Cts (Cycle Thresholds [for the PCR]) likely due to bubble formation and insufficient mixing during the procedure run. The mouse liver qRT-PCR run had successful singleplex and duplex reactions and the variability was slightly better as the mixing operation was improved. The ability to purify and stabilize RNA and to conduct qRT-PCR on-orbit is an important step towards utilizing the ISS as a National Laboratory facility. The ability to get on-orbit data will provide investigators with the opportunity to adjust experimental parameters in real time without the need for sample return and re-flight. The WetLab-2 Project is supported by the Research Integration Office in the ISS Program.

gene expression↗
NASA NTRS2016

Successful Validation of Sample Processing and Quantitative Real-Time PCR Capabilities on the International Space Station

The WetLab-2 system was developed by NASA Ames Research Center to offer new capabilities to researchers. The system can lyse cells and extract RNA (Ribonucleic Acid) on-orbit from different sample types ranging from microbial cultures to animal tissues. The purified RNA can then either be stabilized for return to Earth or can be used to conduct on-orbit quantitative Reverse Transcriptase PCR (Polymerase Chain Reaction) (qRT-PCR) analysis without the need for sample return. The qRT-PCR results can be downlinked to the ground a few hours after the completion of the run. The validation flight of the WetLab-2 system launched on SpaceX-8 on April 8, 2016. On orbit operations started on April 15th with system setup and was followed by three quantitative PCR runs using an E. coli genomic DNA template pre-loaded at three different concentrations. These runs were designed to discern if quantitative PCR functions correctly in microgravity and if the data is comparable to that from the ground control runs. The flight data showed no significant differences compared to the ground data though there was more variability in the values, this was likely due to the numerous small bubbles observed. The capability of the system to process samples and purify RNA was then validated using frozen samples prepared on the ground. The flight data for both E. coli and mouse liver clearly shows that RNA was successfully purified by our system. The E. coli qRT-PCR run showed successful singleplex, duplex and triplex capability. Data showed high variability in the resulting Cts (Cycle Thresholds [for the PCR]) likely due to bubble formation and insufficient mixing during the procedure run. The mouse liver qRT-PCR run had successful singleplex and duplex reactions and the variability was slightly better as the mixing operation was improved. The ability to purify and stabilize RNA and to conduct qRT-PCR on-orbit is an important step towards utilizing the ISS as a National Laboratory facility. The ability to get on-orbit data will provide investigators with the opportunity to adjust experimental parameters in real time without the need for sample return and re-flight. The WetLab-2 Project is supported by the Research Integration Office in the ISS Program.

quantitative PCR↗
DOE OSTI2025

New approaches to secondary metabolite discovery from anaerobic gut microbes

The animal gut microbiome is a complex system of diverse, predominantly anaerobic microbiota with secondary metabolite potential. These metabolites likely play roles in shaping microbial community membership and influencing animal host health. As such, novel secondary metabolites from gut microbes hold significant biotechnological and therapeutic interest. Despite their potential, gut microbes are largely untapped for secondary metabolites, with gut fungi and obligate anaerobes being particularly under-explored. To advance understanding of these metabolites, culture-based and (meta)genome-based approaches are essential. Culture-based approaches enable isolation, cultivation, and direct study of gut microbes, and (meta)genome-based approaches utilize in silico tools to mine biosynthetic gene clusters (BGCs) from microbes that have not yet been successfully cultured. In this mini-review, we highlight recent innovations in this area, including anaerobic biofoundries like ExFAB, the NSF BioFoundry for Extreme & Exceptional Fungi, Archaea, and Bacteria. These facilities enable high-throughput workflows to study oxygen-sensitive microbes and biosynthetic machinery. Such recent advances promise to improve our understanding of the gut microbiome and its secondary metabolism.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2025

Volatiles from the necrophagous fly Cochliomyia macellaria (Diptera: Calliphoridae) as indicators of Salmonella exposure

Blow flies (Diptera: Calliphoridae) are crucial in forensic investigations due to their association with both living and dead humans and other animals. Additionally, their interactions with various resources and potential as vectors of pathogens of humans and other animals, thus, make them potential tools for biosurveillance. This study investigated the potential of monitoring volatile organic compounds (VOCs) emitted by blow flies exposed to Salmonella as a method for pathogen surveillance. Adult blow flies ( Cochliomyia macellaria ) were exposed, or not, to Salmonella enterica . Following exposure, VOCs released by the blow flies were collected and analyzed using gas chromatography-mass spectrometry (GC-MS). Results indicate a treatment by time interaction (P < 0.01). Indicator species analysis identified a single compound significantly associated with S. enterica exposure (P = 0.02), Nonane, 2,2,4,4,6,8,8-heptamethyl, potentially indicating an immune system response. Given a compound indicating exposure was detected, future research should determine if more replicates could detect more differences after Salmonella ingestion. This research highlights the potential of blow flies as biosurveillance tools and the potential value of volatiles for assessing their exposure to pathogens.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2016

The life science x-ray scattering beamline at NSLS-II

We report the current development status of the High Brightness X-ray Scattering for Life Sciences (or Life Science X-ray Scattering, LiX) beamline at the NSLS-II facility of Brookhaven National Laboratory. This instrument will operate in the x-ray energy range of 2.1-18 keV, provide variable beam sizes from 1 micron to ∼0.5 mm, and support user experiments in three scientific areas: (1) high-throughput solution scattering, in-line size exclusion chromatography and flow mixers-based time-resolved solution scattering of biological macro-molecules, (2) diffraction from single- and multi-layered lipid membranes, and (3) scattering-based scanning probe imaging of biological tissues. In order to satisfy the beam stability required for these experiments and to switch rapidly between different types of experiments, we have adopted a secondary source with refractive lenses for secondary focusing, a detector system consisting of three Pilatus detectors, and specialized experimental modules that can be quickly exchanged and each dedicated to a defined set of experiments. The construction of this beamline is on schedule for completion in September 2015. User experiments are expected to start in Spring 2016.

43 PARTICLE ACCELERATORS↗
NASA NTRS1990

Activity and adaptation of nitrilotriacetate (NTA)-degrading bacteria: field and laboratory studies

Adaptation of bacterial activity for the degradation of nitrilotriacetate (NTA) was studied using natural sediment samples and an NTA-degrading bacterium (strain ATCC 29600). Sediment samples from a river with persistent levels of NTA had much higher NTA-degradative activity than comparable samples from a less contaminated control site. When sediment from the control site was exposed to high levels of NTA a 5 day lag preceded an abrupt increase in NTA degradation while strain 29600 colonized on sand and grown in the absence of NTA became induced within eight hours. The induction of strain 29600 was compared between bacteria in suspension and cells attached to sand. The sand-associated bacteria became induced 4 to 5 h before the planktonic suspension and displayed over threefold greater specific activity. Suspensions of strain 29600 became adapted within 8 h when placed in membrane diffusion chambers that were immersed within a municipal wastewater reactor containing NTA. These findings support the concept that induction is a part of the process of bacterial adaptation to degrade NTA and sand-associated bacteria can adapt more quickly to and have a greater degradative activity for NTA than planktonic cells.

NASA Discipline Number 04-10↗
DOE OSTI2010

Melanin-Covered Nanoparticles for Protection of Bone Marrow During Radiation Therapy of Cancer

Purpose: Protection of bone marrow against radiotoxicity during radioimmunotherapy and in some cases external beam radiation therapy such as hemi-body irradiation would permit administration of significantly higher doses to tumors, resulting in increased efficacy and safety of treatment. Melanin, a naturally occurring pigment, possesses radioprotective properties. We hypothesized that melanin, which is insoluble, could be delivered to the bone marrow by intravenously administrated melanin-covered nanoparticles (MNs) because of the human body's 'self-sieving' ability, protecting it against ionizing radiation. Methods and Materials: The synthesis of MNs was performed via enzymatic polymerization of 3,4-dihydroxyphenylalanine and/or 5-S-cysteinyl-3,4-dihydroxyphenylalanine on the surface of 20-nm plain silica nanoparticles. The biodistribution of radiolabeled MNs in mice was done at 3 and 24 h. Healthy CD-1 mice (Charles River Laboratories International, Inc., Wilmington, MA) or melanoma tumor-bearing nude mice were given MNs intravenously, 50 mg/kg of body weight, 3 h before either whole-body exposure to 125 cGy or treatment with 1 mCi of {sup 188}Re-labeled 6D2 melanin-binding antibody. Results: Polymerization of melanin precursors on the surface of silica nanoparticles resulted in formation of a 15-nm-thick melanin layer as confirmed by light scattering, transmission electron microscopy, and immunofluorescence. The biodistribution after intravenous administration showed than MN uptake in bone marrow was 0.3% and 0.2% of injected dose per gram at 3 and 24 h, respectively, whereas pre-injection with pluronic acid increased the uptake to 6% and 3% of injected dose per gram, respectively. Systemic MN administration reduced hematologic toxicity in mice treated with external radiation or radioimmunotherapy, whereas no tumor protection by MNs was observed. Conclusions: MNs or similar structures provide a novel approach to protection of bone marrow from ionizing radiation based on prevention of free radical formation by melanin.

62 RADIOLOGY AND NUCLEAR MEDICINE↗
NASA NTRS1992

Chromosomal changes in cultured human epithelial cells transformed by low- and high-LET radiation

For a better assessment of radiation risk in space, an understanding of the responses of human cells, especially the epithelial cells, to low- and high-LET radiation is essential. In our laboratory, we have successfully developed techniques to study the neoplastic transformation of two human epithelial cell systems by ionizing radiation. These cell systems are human mammary epithelial cells (H184B5) and human epidermal keratinocytes (HEK). Both cell lines are immortal, anchorage dependent for growth, and nontumorigenic in athymic nude mice. Neoplastic transformation was achieved by irradiating cells successively. Our results showed that radiogenic cell transformation is a multistep process and that a single exposure of ionizing radiation can cause only one step of transformation. It requires, therefore, multihits to make human epithelial cells fully tumorigenic. Using a simple karyotyping method, we did chromosome analysis with cells cloned at various stages of transformation. We found no consistent large terminal deletion of chromosomes in radiation-induced transformants. Some changes of total number of chromosomes, however, were observed in the transformed cells. These transformants provide an unique opportunity for further genetic studies at a molecular level.

NASA Discipline Number 04-10↗
NASA NTRS2015

NASA's Rodent Research Project on ISS: Validation of a New Platform for Conducting Biomedical and Basic Research into the Consequences of Long Duration Habitation in Space

Rodent research has played a key role in advancing biomedical discoveries both on Earth and in space. The National Research Counsel’s Decadal survey(1) emphasized the importance of expanding NASAs life sciences research to perform long duration, rodent experiments on the International Space Station (ISS). To accomplish this objective, flight hardware, operations, and science capabilities were developed at NASA ARC to support both commercial and government-sponsored rodent research.Rodent Research-1 (RR-1) was the first mission in which animals were delivered and maintained in the ISS for a long duration mission in modified Animal Enclosure Module hardware. Both RR validation and commercial science objectives were pursued on the RR-1 mission. Adult female mice (20 total Flight, FLT) were launched Sept 21, 2014 in RR hardware within a Dragon Capsule (SpaceX4), then after 4 days in transit, were transferred for habitation on the ISS for 17 days (commercial) or 33 days (validation), when animals were euthanized and select tissues recovered on orbit. Various controls groups consisted of: 1) Basal mice from the same cohorts as FLT mice, but tissues were recovered at time of launch, 2) Vivarium (VIV) were housed in standard cages 3) Ground Controls (GC) were housed in flight hardware within an environmental chamber at Kennedy Space Center. The health and behavior of all mice on the ISS were monitored by video feed on a daily basis. Mice were euthanized by injection of Euthasol, then either fast frozen intact or dissected to preserve livers (fast frozen) and spleens (RNAlater). Samples were stored at ≤ -80˚C until their return to Earth for later analyses.Hardware performed nominally throughout the mission and the planned in-flight science operations were completed successfully. FLT mice appeared generally more physically active on orbit than respective GC groups. After 33 days on the ISS, mean body weights of FLT mice did not differ from GC, with both groups showing a 6% rise compared to time of launch, while VIV mice showed an 8% rise over the same period. Importantly, there were no significant differences in body weights between groups at the end of 33 days on the ISS, providing an indication that the RR hardware supported the health of the mice both on Earth and in space. Based on the preliminary data obtained from the livers and spleens of mice after 17 days on the ISS, purified RNA was of high quality (RIN values of spleen: FLT=9.48 +0.40, GC=9.28 +0.44, n=5/group); therefore, RNA quality from samples retrieved on orbit was acceptable for even the most demanding transcriptomic analyses. In addition, liver enzyme activity levels (units/mg protein) of FLT mice (after 17d on ISS) and all control mice were similar in magnitude to samples that were optimally prepared by freezing in liquid nitrogen in the laboratory (enzymes analyzed included catalase, glutathione reductase and glyceraldehyde-3-phosphate dehydrogenase). Validation analyses still in progress include behavior and tissue biochemistries, as well as optimization of science return by post-flight recovery of tissues for biospecimen sharing and global expression analyses.Together, these preliminary findings demonstrate new capability for supporting long duration rodent research on the ISS to achieve both basic science and biomedical objectives.

Globus, R. K.↗
DOE OSTI2025

Biomass-Derived Carbon and Their Composites for Supercapacitor Applications: Sources, Functions, and Mechanisms

Biomass-derived carbons are eco-friendly and sustainable materials, making them ideal for supercapacitors due to their high surface area, excellent conductivity, cost-effectiveness, and environmental benefits. This review provides valuable insights into biomass-derived carbon and modified carbon for supercapacitors, integrating both experimental results and theoretical calculations. This review begins by discussing the origins of biomass-derived carbon in supercapacitors, including plant-based, food waste-derived, animal-origin, and microorganism-generated sources. Then, this review presents strategies to improve the performance of biomass-derived carbon in supercapacitors, including heteroatom doping, surface functionalization, and hybrid composite construction. Furthermore, this review analyzes the functions of biomass-derived carbon in supercapacitors both in its pure form and as modified materials. The review also explores composites derived from biomass-based carbon, including carbon/MXenes, carbon/MOFs, carbon/graphene, carbon/conductive polymers, carbon/transition metal oxides, and carbon/hydroxides, providing a thorough investigation. Most importantly, this review offers an innovative summary and analysis of the role of biomass-derived carbon in supercapacitors through theoretical calculations, concentrating on four key aspects: energy band structure, density of states, electron cloud density, and adsorption energy. Finally, the review concludes the future research directions for biomass carbon-based supercapacitors, including the discovery of novel biomass materials, tailoring surface functional groups, fabricating high-performance composite materials, exploring ion transfer mechanisms, and enhancing practical applications. In summary, this review offers a thorough exploration of the sources, functions, and mechanisms of biomass-derived carbon in supercapacitors, providing valuable insights for future research.

biomass-derived carbon↗
DOE OSTI2015

Multipurpose epithermal neutron beam on new research station at MARIA research reactor in Swierk-Poland

MARIA reactor is an open-pool research reactor what gives the chance to install uranium fission converter on the periphery of the core. It could be installed far enough not to induce reactivity of the core but close enough to produce high flux of fast neutrons. Special design of the converter is now under construction. It is planned to set the research stand based on such uranium converter in the near future: in 2015 MARIA reactor infrastructure should be ready (preparation started in 2013), in 2016 the neutron beam starts and in 2017 opening the stand for material and biological research or for medical training concerning BNCT. Unused for many years, horizontal channel number H2 at MARIA research rector in Poland, is going to be prepared as a part of unique stand. The characteristics of the neutron beam will be significant advantage of the facility. High flux of neutrons at the level of 2x10{sup 9} cm{sup -2}s{sup -1} will be obtainable by uranium neutron converter located 90 cm far from the reactor core fuel elements (still inside reactor core basket between so called core reflectors). Due to reaction of core neutrons with converter U{sub 3}Si{sub 2} material it will produce high flux of fast neutrons. After conversion neutrons will be collimated and moderated in the channel by special set of filters and moderators. At the end of H2 channel i.e. at the entrance to the research room neutron energy will be in the epithermal energy range with neutron intensity at least at the level required for BNCT (2x10{sup 9} cm{sup -2}s{sup -1}). For other purposes density of the neutron flux could be smaller. The possibility to change type and amount of installed filters/moderators which enables getting different properties of the beam (neutron energy spectrum, neutron-gamma ratio and beam profile and shape) is taken into account. H2 channel is located in separate room which is adjacent to two other empty rooms under the preparation for research laboratories (200 m2). It is planned to create fully equipped complex facility possible to perform various experiments on the intensive neutron beam. Epithermal neutron beam enables development across the full spectrum of materials research for example shielding concrete tests or electronic devices construction improvement. Due to recent reports on the construction of the accelerator for the Boron Neutron Capture Therapy (BNCT) it has the opportunity to become useful and successful method in the fight against brain and other types of cancers not treated with well known medical methods. In Europe there is no such epithermal neutron source which could be used throughout the year for training and research for scientist working on BNCT what makes the stand unique in Europe. Also our research group which specializes in mixed radiation dosimetry around nuclear and medical facilities would be able to carry out research on new detectors and methods of measurements for radiological protection and in-beam (therapeutic) dosimetry. Another group of scientists from National Centre for Nuclear Research, where MARIA research reactor is located, is involved in research of gamma detector systems. There is an idea to develop Prompt-gamma Single Photon Emission Computed Tomography (Pg- SPECT). This method could be used as imaging system for compounds emitting gamma rays after nuclear reaction with thermal neutrons e.g. for boron concentration in BNCT. Inside the room, where H2 channel is located, there is another horizontal channel - H1 which is also unused. Simultaneously with the construction of the H2 stand it will be possible to create special pneumatic horizontal mail inside the H1 channel for irradiation material samples in the vicinity of the core i.e. in the distal part of the H1 channel. It might expand the scope of research at the planned neutron station. Secondly it is planned to equip both stands with moveable positioning system, video system and facilities to perform animal experiments (anaesthesia, vital signs control, imaging devices, positioning). These all above make constructed station unique in the world (uranium fission converter-based beam) and the only one of such intense neutron beam in the Europe. Moreover implementation of the station would allow the development of research on a number of issues for researchers from all over the Europe. One of very important advantages of the station is undisturbed exploitation of the reactor and other vertical and horizontal channels. MARIA reactor operates 6000 hours per year and that amount of time will be achievable for research on the neutron station. It have to be underlined that new neutron station will work parallel to all another ventures. (authors)

21 SPECIFIC NUCLEAR REACTORS AND ASSOCIATED PLANTS↗
DOE OSTI2017

Flash Pyrolysis of Oleaginous Biomass in a Fluidized-Bed Reactor

In this study, flash pyrolysis was performed using milled soybean as a model substrate to assess the production of liquid fuels from oleaginous biomass feedstocks. A laboratory-scale fluidized-bed flash pyrolysis reactor that allowed rapid heat transfer to the biomass along with short vapor residence time was designed and constructed. Pyrolysis was performed between 250 and 610 °C with a vapor residence time between 0.2 and 0.3 s. At 550 °C or higher, nearly 70% of the initial feed mass as well as feed carbon was recovered in bio-oil. In addition, 90% of the feedstock lipids were recovered in the bio-oil at these pyrolysis conditions. The high liquid products yields were attributed to (1) the low secondary degradation of bio-oils due to the short vapor residence time and (2) the high recovery of liquids in a novel dry ice packed-bed condenser that provided a high surface area for condensation/aggregation of dilute bio-oil vapors/aerosols that were entrained in the carrier gas. The bio-oil from this study had higher C and H content, higher calorific value, and lower oxygen and water content than bio-oil from wood. Finally, these results show that high-quality bio-oil at high yield can be obtained from flash pyrolysis of oleaginous feedstock.

09 BIOMASS FUELS↗
DOE OSTI2025

Plant metacaspases orchestrate wound‐induced pathways for immunity and tissue regeneration

Wounding in plants elicits immunity and tissue repair, but how these responses are coordinated has yet to be elucidated. While plant metacaspases resemble animal caspases in structure and immunity induction, their role in tissue repair and regeneration is unknown. Using Arabidopsis mutants lacking type II metacaspases AtMC4 or AtMC9, we found that the majority of the highly induced, wound-responsive genes in Arabidopsis thaliana are suppressed by the loss of AtMC4, while AtMC9 plays an auxiliary role in defense activation. Specifically, AtMC4, but not AtMC9, is required for the activation of genes involved in tissue repair, such as the developmental regulator WOX5, as well as for root regeneration from excised leaves. Instead, AtMC9 mediates the repression of a subset of basal immunity genes, which modifies the wound-activated defense response from that induced by molecular patterns such as the bacterial flg22 elicitor. Our results thus reveal a conserved protease module that coordinates plant defense and tissue repair upon wounding. They could be new targets to improve crop performance and plant transformation protocols that involve tissue wounding before transgenic plant selection and regeneration. The groups of genes with distinctive requirements for the two metacaspases could provide markers to dissect how these specialized proteases affect different response pathways that underpin the multifaceted wounding response.

54 ENVIRONMENTAL SCIENCES↗
NASA NTRS1992

Utilization of Space Station Freedom for technology research

Space Station Freedom presents a unique opportunity for technology developers to conduct research in the space environment. Research can be conducted in the pressurized volume of the Space Station's laboratories or attached to the Space Station truss in the vacuum of space. Technology developers, represented by the Office of Aeronautics and Space Technology (OAST), will have 12 percent of the available Space Station resources (volume, power, data, crew, etc.) to use for their research. Most technologies can benefit from research on Space Station Freedom and all these technologies are represented in the OAST proposed traffic model. This traffic model consists of experiments that have been proposed by technology developers but not necessarily selected for flight. Experiments to be flown in space will be selected through an Announcement of Opportunity (A.O.) process. The A.O. is expected to be released in August, 1992. Experiments will generally fall into one of the 3 following categories: (1) Individual technology experiments; (2) Instrumented Space Station; and (3) Guest investigator program. The individual technology experiments are those that do not instrument the Space Station nor directly relate to the development of technologies for evolution of Space Station or development of advanced space platforms. The Instrumented Space Station category is similar to the Orbiter Experiments Program and allows the technology developer to instrument subsystems on the Station or develop instrumentation packages that measure products or processes of the Space Station for the advancement of space platform technologies. The guest investigator program allows the user to request data from Space Station or other experiments for independent research. When developing an experiment, a developer should consider all the resources and infrastructure that Space Station Freedom can provide and take advantage of these to the maximum extent possible. Things like environment, accommodations, carriers, and integration should all be taken into account. In developing experiments at Langley Research Center, an iterative approach is proving useful. This approach uses Space Station utilization and subsystem experts to advise and critique experiment designs to take advantage of everything the Space Station has to offer. Also, solid object modeling and animation computer tools are used to fully visualize the experiment and its processes. This process is very useful for attached payloads and allows problems to be detected early in the experiment design phase.

Avery, Don E.↗
DOE OSTI2025

γ-Aminobutyric acid modulates terpene biosynthesis through the ATG8a-mediated pathway

Terpenes play crucial roles in plant growth, development, and stress responses. The biosynthesis of terpenes is influenced by abiotic stress factors, such as drought, temperature, or light. Here, however, the molecular network underlying how terpenes are regulated in response to environmental stimuli remains largely unknown. Here, we identified the autophagy protein SgATG8a as a key mediator of GABA-regulated terpene production and drought tolerance in Sindora glabra. SgATG8a, evolutionarily related to the animal GABA receptor-associated protein (GABARAP) subfamily, localizes in both the nucleus and cytoplasm. Exogenous GABA treatment not only increased the expression level of terpene synthase genes (SgTPSs) but also led to enhanced accumulation of six main terpene components in Sindora glabra. In addition, GABA alleviated the photosynthesis damage and enhanced leaf biomass under drought conditions. Consistently, overexpression of SgATG8a in Arabidopsis increased terpene synthase gene (SgTPS) expression, leading to the enhanced production of four major terpenes and improved the tolerance of transgenic plants to drought stress by regulating reactive oxygen species (ROS) scavenging systems. Moreover, the transcription factors SgWRKY13 and SgERF4 were identified as interacting partners of SgATG8a, activating SgTPS3 expression. Lectin receptor-like kinase (LecRK1) is involved in the GABA-mediated pathway by interacting with the SgWRKY13/SgERF4-SgATG8a proteins, and the LecRK1-SgWRKY13/SgERF4 phosphorylation module fine-tunes the transcription of the downstream SgTPS3 gene. Taken together, these findings reveal a novel role for GABA in regulating terpene biosynthesis and drought tolerance, providing insights into the molecular mechanism underlying GABA-mediated terpene production.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2016

The MELCOR Analysis of Chinshan Nuclear Power Plant Spent Fuel Pool for Fukushima-like Accident

The safety analysis of the nuclear power plant (NPP) is very important work in the NPP safety. Especially after the Fukushima NPP accident occurred, the importance of NPP safety analysis has been raised and there is more concern for the safety of NPPs in Taiwan. Because the earthquake and tsunami occurred, the cooling system of spent fuel pool failed and the safety issue of spent fuel pool generated in Japan's Fukushima NPP. Chinshan NPP is the first NPP in Taiwan which is BWR/4 plant and OLTP (Original Licensed Thermal Power) for each unit is 1775 MWt. Chinshan NPP finished SPU (stretch power uprate) and the operating power is 103.66% of the OLTP, which is 1840 MWt now. After Fukushima NPP accident occurred, in order to concern the safety of Chinshan NPP spent fuel pool, we performed the safety analysis of spent fuel pool by using MELCOR/SNAP which also assumed the cooling system of spent fuel pool failed. The geometry of Chinshan NPP spent fuel pool is 12.17 m x 7.87 m x 11.61 m and the initial condition is 60 deg. (water temperature) / 1.013 x 10{sup 5} Pa. And, the total power of the fuels is roughly 8.9 MWt initially. MELCOR is developed by Sandia National Laboratories. MELCOR is a fully integrated, engineering-level computer code that models the progression of severe accidents in light water reactor NPPs, including containment response and leakage of activity to the environment. A broad spectrum of severe accident phenomena in both BWRs and PWRs is treated in MELCOR in a unified framework. These include thermalhydraulic response in the reactor coolant system, reactor cavity, containment, and confinement buildings; core heatup, degradation, and relocation; core-concrete attack; hydrogen production, transport, and combustion; fission product release and transport behavior. Current uses of MELCOR include estimation of severe accident source terms and their sensitivities and uncertainties in a variety of applications. According to Dr. Carbajo's paper, MELCOR was used to perform the study of spent fuel pool of Fukushima Daiichi Unit 4. Niina E. Koenoenen used MELCOR to establish the spent fuel pool model of a Nordic BWR. The loss-of-pool-cooling accidents was simulated and analyzed by this model. The above studies indicate that MELCOR is capable of handling the simulation of the spent fuel pool. An increasing number of researchers are using MELCOR code to analyze test facilities and nuclear power plants. Jun Wang et al. used MELCOR to perform core degradation simulation and MELCOR results were compared with CORA experimental data. Through this work, they reviewed the performance of MELCOR COR package in detail. Tae Woon Kim et al. developed MELCOR model of APR1400 (an advanced pressurized water reactor). They used this model to perform the LBLOCA transient simulation and sensitivity study. Longze Li et al. established the MELCOR model of CPR1000 (a Chinese pressurized reactor 1000-MW power plant). The station blackout (SBO) with failure of the steam generator (SG) safety relief valve (SRV) transient was simulated and analyzed by this model. SNAP is a graphic user interface program that processes the inputs and outputs of MELCOR. In addition, there is an animation function in SNAP which can present the animation of analysis results. There are two main steps in this research. The first step was the establishment of Chinshan NPP spent fuel pool MELCOR/SNAP model. And the transient analysis of Chinshan NPP spent fuel pool MELCOR/SNAP model under the cooling system failure condition was performed. The next step was the comparison among the MELCOR, TRACE and CFD data. TRACE and CFD data were from INER reports. In addition, the animation model of Chinshan NPP spent fuel pool was presented by using the animation function of SNAP with MELCOR/SNAP analysis results. This study has developed MELCOR/SNAP model of Chinshan NPP spent fuel pool successfully. By using the above model, the safety analysis of the spent fuel pool was performed under the cooling system of spent fuel pool failed condition. The analysis results of MELCOR, TRACE and CFD were similar in this case. It indicated that there was a respectable accuracy in MELCOR/SNAP model. The analysis results depicted that the uncovered of the fuels occurred at 2.6 day and the metal-water reaction of fuels occurred roughly at 3.7 day after the cooling system failed. The above results indicated that the failure of cladding occurred after 3.7 day. This study's results can help to evaluate the safety issue of Chinshan NPP spent fuel pool. In addition, the cooling system and some mitigation measures of spent fuel pool is not simulated in the above model. However, we will do it in the future. (authors)

21 SPECIFIC NUCLEAR REACTORS AND ASSOCIATED PLANTS↗
DOE OSTI2025

BONCAT-Live for isolation and cultivation of active environmental bacteria

In diverse environments, microbes drive a myriad of processes, from geochemical and nutrient cycling to interspecies interactions, including associations with plants and animals. Their physiological state is dynamic and impacted by abiotic and biotic conditions, responding to environmental fluctuations by changes in cellular metabolism, according to their genetic potential. Molecular, cellular, and genomic approaches can identify and measure microbial responses and adaptation to environmental changes in native communities. However, isolating individual microbial cells that respond to specific changes for cultivation has been difficult. To address this, we implemented a novel bacterial isolation approach (BONCAT-Live) by integrating bio-orthogonal non-canonical amino acid tagging (BONCAT) in diverse native communities, with isolation and cultivation of cells responding to specific stimuli, at different time scales. In frozen Arctic permafrost samples, we identified and isolated dormant bacteria that become active after thawing under native or nutrient-enriched conditions. From the Populus tree rhizosphere, we isolated strains that thrive under high concentrations of root exudates that act as defense compounds and nutrients. In the human microbiome, we identified and isolated bacteria that rapidly proliferated when exposed to metabolites provided by the host or other co-occurring microbes. Further characterization of isolated bacterial strains will provide opportunities for in-depth determination of how these microbes adapt to changes in their environments, individually and as part of model communities.

Analytical Methods↗
NASA NTRS1996

Gravitational physiology of human immune cells: a review of in vivo, ex vivo and in vitro studies

The study of the function of immune cells in microgravity has been studied for more than 20 years in several laboratories. It is clear today that the immune system is depressed in more than 50% of the astronauts during and after space flight and that the activation of T lymphocytes by mitogens in vitro changes dramatically. This article gives an overview of the gravitational studies conducted by our laboratory in Spacelab, in MIR station, in sounding rockets and on the ground in the clinostat and the centrifuge. Three experimental approaches are followed in our work: (i) Ex vivo studies are performed with blood samples drawn from astronauts; (ii) in vivo studies are based on the application of seven antigens to the skin of the astronauts; (iii) in vitro studies are carried out with immune cells purified from the blood of healthy donors (not astronauts). The data from our in vivo and ex vivo studies are in agreement with those of other laboratories and show that the immunological function is depressed in the majority of astronauts as a consequence of the stress of space flight rather than by a direct influence of gravity on the cell. Immune depression may become a critical hazard on long duration flights on space stations or to other planets. In vitro experiments show that cultures of free-floating lymphocytes and monocytes undergo a dramatic depression of activation by the mitogen concanavalin A, while activation is more than doubled when the cells are attached to microcarrier beads. Such effects may be attributed to both direct and indirect effects of gravitational unloading on basic biological mechanisms of the cell. While the in vitro data are very important to clarify certain aspects of the biological mechanism of T cells activation, they are not descriptive of the changes of the immunological function of the astronauts.

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