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Detection of DNA Microsatellites Using Multiplex Polymerase Chain Reaction Aboard the International Space Station 利用多重聚合酶链反应在国际空间站上检测DNA微卫星
Sophia Chen, J. Hatch, A. Luck, N. Nichols, Emily J Gleason, K. Martin, Kevin D. Foley, D. Scott Copeland, Sebastian Kraves, E. Saavedra
Abstract As human exploration extends further into deep space, it is critical to understand the cellular impacts of spaceflight in order to ensure the safety of future astronauts. Extended exposure to cosmic radiation and microgravity has been shown to cause genetic damage and impair cellular DNA repair mechanisms, which together can lead to genomic instability. In particular, microsatellite instability (MSI), in which dysfunction in DNA mismatch repair (MMR) causes alterations in tandemly repeated “microsatellite” sequences, is a manifestation of genomic instability that has been associated with certain cancers. In this study, we establish the feasibility of an on-orbit multiplex polymerase chain reaction (PCR)-based assay to detect mutations in cancer-related microsatellites. Multiplex PCR was used to amplify five quasimonomorphic microsatellites in space and on Earth from both wild-type and MMR-deficient human cell lines. These data provide proof of concept of simultaneous amplification of multiple DNA sequences in space, expanding in-flight research and health-monitoring capabilities.
随着人类探索进一步深入太空,了解太空飞行对细胞的影响对于确保未来宇航员的安全至关重要。长期暴露在宇宙辐射和微重力下已被证明会造成遗传损伤和损害细胞DNA修复机制,两者共同导致基因组不稳定。特别是,微卫星不稳定性(MSI),即DNA错配修复(MMR)功能障碍导致串联重复的“微卫星”序列改变,是与某些癌症相关的基因组不稳定性的一种表现。在这项研究中,我们建立了一种基于在轨多重聚合酶链反应(PCR)的检测癌症相关微卫星突变的可行性。利用多重PCR技术,从野生型和mmr缺陷的人类细胞系中扩增出5个准单态微卫星。这些数据证明了在太空中同时扩增多个DNA序列的概念,扩大了飞行中研究和健康监测能力。
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引用次数: 0
Shared Metabolic Remodeling Processes Characterize the Transcriptome of Arabidopsis thaliana within Various Suborbital Flight Environments 不同亚轨道飞行环境下拟南芥共享代谢重塑过程的转录组特征
Brandon Califar, Agata K. Zupanska, Jordan A. Callaham, M. Bamsey, T. Graham, A. Paul, R. Ferl
Abstract The increasing availability of flights on suborbital rockets creates new avenues for the study of spaceflight effects on biological systems, particularly of the transitions between hypergravity and microgravity. This paper presents an initial comparison of the responses of Arabidopsis thaliana to suborbital and atmospheric parabolic flights as an important step toward characterizing these emerging suborbital platforms and their effects on biology. Transcriptomic profiling of the response of the Arabidopsis ecotype Wassilewskija (WS) to the aggregate suborbital spaceflight experiences in Blue Origin New Shepard and Virgin Galactic SpaceShipTwo revealed that the transcriptomic load induced by flight differed between the two flights, yet was biologically related to traditional parabolic flight responses. The sku5 skewing mutant and 14-3-3κ:GFP regulatory protein overexpression lines, flown in the Blue Origin and parabolic flights, respectively, each showed altered intra-platform responses compared to WS. An additional parabolic flight using the F-104 Starfighter showed that the response of 14-3-3κ:GFP to flight was modulated in a similar manner to the WS line. Despite the differing genotypes, experimental workflows, flight profiles, and platforms, differential gene expression linked to remodeling of central metabolic processes was commonly observed in the flight responses. However, the timing and directionality of differentially expressed genes involved in the conserved processes differed among the platforms. The processes included carbon and nitrogen metabolism, branched-chain amino acid degradation, and hypoxic responses. The data presented herein highlight the potential for various suborbital platforms to contribute insights into biological responses to spaceflight, and further suggest that in-flight fixation during suborbital experiments will enhance insights into responses during each phase of flight.
亚轨道火箭飞行的不断增加为研究太空飞行对生物系统的影响,特别是超重力和微重力之间的过渡提供了新的途径。本文介绍了拟南芥对亚轨道和大气抛物线飞行的反应的初步比较,作为表征这些新兴亚轨道平台及其对生物学影响的重要一步。拟南芥生态型Wassilewskija (WS)对蓝色起源新谢泼德号和维珍银河太空船2号亚轨道飞行经历的转录组学分析显示,两次飞行诱导的转录组负荷不同,但与传统的抛物线飞行反应具有生物学相关性。与WS相比,分别在蓝色起源和抛物线飞行中飞行的sku5倾斜突变体和14-3-3κ:GFP调节蛋白过表达系均显示出平台内响应的改变。使用F-104 Starfighter进行的额外抛物线飞行表明,14-3-3κ:GFP对飞行的响应以与WS线相似的方式被调制。尽管存在不同的基因型、实验工作流程、飞行概况和平台,但在飞行反应中普遍观察到与中枢代谢过程重塑相关的差异基因表达。然而,参与保守过程的差异表达基因的时间和方向性在不同的平台上是不同的。这一过程包括碳氮代谢、支链氨基酸降解和缺氧反应。本文提供的数据强调了各种亚轨道平台有助于了解太空飞行的生物反应的潜力,并进一步表明,亚轨道实验期间的飞行固定将增强对飞行每个阶段反应的了解。
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引用次数: 3
Liquid Propellant Mass Measurement in Microgravity 微重力下液体推进剂质量测量
K. Crosby, R. Werlink, E. Hurlbert
Abstract The Modal Propellant Gauging (MPG) experiment has demonstrated sub-1% gauging accuracy under laboratory conditions on both flight hardware and subscale tanks. Recently, MPG was adapted for flight on Blue Origin's New Shepard vehicle and has flown twice, achieving equilibrated, zero-g surface configurations of propellant simulant at three different fill fractions. Flight data from MPG missions on New Shepard P7 and P9 show agreement between known and measured propellant levels of 0.3% for the fill fractions investigated in the present study. Two approaches for estimating zero-g propellant mass are described here. Both approaches rely on measuring shifts in modal frequencies of a tank excited by acoustic surface waves and subject to fluid mass loading by the propellant. In the first approach, shifts in the lowest mode frequency (LMF) are measured and associated with liquid fill-level changes. In the second approach, 1-g modal spectra at a range of known fill levels are used in a cross-correlation calculation to predict fill levels associated with a zero-g modal spectrum. Flight data for both approaches are consistent with finite element predictions using a simple fluid–structure interaction model. In both settled and unsettled microgravity environments, MPG meets or exceeds NASA Roadmap goals for in-space propellant mass gauging.
模态推进剂量测(MPG)实验表明,在实验室条件下,飞行硬件和亚尺度油箱的量测精度都低于1%。最近,MPG在蓝色起源公司的新谢泼德飞行器上进行了改装,并进行了两次飞行,在三种不同的填充分数下实现了推进剂模拟剂的平衡、零重力表面配置。新谢泼德P7和P9上MPG任务的飞行数据表明,在本研究中调查的填充分数中,已知和测量的推进剂水平之间的一致性为0.3%。本文描述了估算零重力推进剂质量的两种方法。这两种方法都依赖于测量由声表面波激发的储罐模态频率的变化,并受推进剂的流体质量载荷的影响。在第一种方法中,测量最低模态频率(LMF)的移位,并将其与液体液位变化相关联。在第二种方法中,在已知填充水平范围内的1g模态光谱用于相互关联计算,以预测与零g模态光谱相关的填充水平。两种方法的飞行数据都与使用简单流固相互作用模型的有限元预测相一致。在稳定和不稳定的微重力环境下,MPG达到或超过了NASA的太空推进剂质量测量路线图目标。
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引用次数: 2
Toward the Analysis of Lymphocyte Development in Space: PCR-Based Amplification of T-Cell Receptor Excision Circles (TRECs) Aboard the International Space Station 太空中淋巴细胞发育的分析:国际空间站上基于pcr的t细胞受体切除环(trec)扩增
Elizabeth Reizis, D. Cai, Lee Serpas, Emily J Gleason, K. Martin, Kevin D. Foley, D. S. Copeland, Sebastian Kraves, E. Saavedra
Abstract Spaceflight offers vast possibilities for expanding human exploration, whereas it also bears unique health risks. One of these risks is immune dysfunction, which can result in the reactivation of latent pathogens and increased susceptibility to infections. The ability to monitor the function of the immune system is critical for planning successful long-term space travel. T lymphocytes are immune cells that develop in the thymus and circulate in the blood. They can detect foreign, infected, or cancerous cells through T cell receptors (TCRs). The assembly of TCR gene segments, to produce functional TCR genes, can be monitored by measuring the presence of TCR excision circles (TRECs), circular fragments of DNA that are by-products of this assembly process mediated by the V(D)J recombination machinery. In this study, we used polymerase chain reaction (PCR) on the International Space Station (ISS) to detect TRECs in murine peripheral blood. We were able to detect TRECs in the blood of normal healthy mice of different ages, with an efficiency comparable to that achieved in ground controls. As expected, we were unable to detect TRECs in the blood of immunodeficient mice. These results are the first step in optimizing a specific, rapid, safe, and cost-effective PCR-based assay to measure the integrity of mammalian immune systems during spaceflight.
太空飞行为人类探索提供了巨大的可能性,同时也承担着独特的健康风险。其中一个风险是免疫功能障碍,这可能导致潜伏病原体的重新激活和对感染的易感性增加。监测免疫系统功能的能力对于规划成功的长期太空旅行至关重要。T淋巴细胞是在胸腺中发育并在血液中循环的免疫细胞。它们可以通过T细胞受体(tcr)检测外来细胞、感染细胞或癌细胞。TCR基因片段的组装,以产生功能性的TCR基因,可以通过测量TCR切除环(TRECs)的存在来监测,TCR切除环是由V(D)J重组机制介导的组装过程的副产物。在本研究中,我们利用国际空间站(ISS)上的聚合酶链反应(PCR)检测小鼠外周血中的trec。我们能够在不同年龄的正常健康小鼠的血液中检测到trec,其效率与地面对照组相当。正如预期的那样,我们无法在免疫缺陷小鼠的血液中检测到trec。这些结果是优化一种特异性、快速、安全和具有成本效益的基于pcr的测定方法的第一步,该方法可用于测量太空飞行期间哺乳动物免疫系统的完整性。
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引用次数: 1
Space Flight Cultivation for Radish (Raphanus sativus) in the Advanced Plant Habitat 先进植物生境中萝卜的航天栽培
S. John, Farid Abou-Issa, K. Hasenstein
Abstract In preparation of a flight experiment, ground-based studies for optimizing the growth of radishes (Raphanus sativus) were conducted at the ground-based Advanced Plant Habitat (APH) unit at the Kennedy Space Center (KSC), Florida. The APH provides a large, environmentally controlled chamber that has been used to grow various plants, such as Arabidopsis, wheat, peppers, and now radish. In support of National Aeronautics and Space Administration (NASA)'s goals to provide astronauts with fresh vegetables and fruits in a confined space, it is important to extend the cultivation period to produce substantial biomass. We selected Raphanus sativus cv. Cherry Belle as test variety both for preliminary tests and flight experiments because it provides edible biomass in as few as four weeks, has desirable secondary metabolites (glucosinolates), is rich in minerals, and requires relatively little space. We report our strategies to optimize the growth substrate, watering regimen, light settings, and planting design that produces good-sized radishes, minimizes competition, and allows for easy harvesting. This information will be applicable for growth optimization of other crop plants that will be grown in the APH or other future plant growth facilities.
为了准备飞行实验,在佛罗里达州肯尼迪航天中心(KSC)的地面先进植物栖息地(APH)单元进行了优化萝卜(Raphanus sativus)生长的地面研究。APH提供了一个大型的环境控制室,用于种植各种植物,如拟南芥、小麦、辣椒,现在是萝卜。为了支持美国国家航空航天局(NASA)在密闭空间内为宇航员提供新鲜蔬菜和水果的目标,延长栽培期以生产大量生物质是很重要的。我们选择Raphanus sativus cv。樱桃百丽作为初步试验和飞行试验的试验品种,因为它在短短四周内提供可食用的生物质,具有理想的次级代谢物(硫代葡萄糖苷),富含矿物质,并且需要相对较少的空间。我们报告了优化生长基质、浇水方案、光照设置和种植设计的策略,以生产出大小良好的萝卜,最大限度地减少竞争,并允许易于收获。该信息将适用于在APH或其他未来植物生长设施中种植的其他作物植物的生长优化。
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引用次数: 3
Using Tapered Channels to Improve LAD Performance for Cryogenic Fluids: Suborbital Testing Results 使用锥形通道改善低温流体的LAD性能:亚轨道测试结果
K. Supak, S. Green, A. McCleney
Abstract Improvement of cryogenic fluid storage and transfer technology for in-space propulsion and storage systems is required for long-term space missions. Screened channel liquid acquisition devices (LADs) have long been used with storable propellants to deliver vapor-free liquid during engine restart and liquid transfer processes. The use of LADs with cryogenic fluids is problematic due to low temperatures associated with cryogenic fluids. External heat leaks will cause vapor bubbles to form within the LADs that are difficult to remove in the existing designs. A tapered LAD channel has been proposed to reliably remove vapor bubbles within the device without costly thrusting maneuvers or active separation systems. A model has been developed to predict bubble movement within tapered LAD channels, and subsequent ground testing was completed with a simulant fluid to provide model validation data. Suborbital microgravity testing of tapered LAD technology was recently completed with two different simulant fluids and demonstrated that the concept can passively expel vapor bubbles within the channel. Two additional suborbital flights have been funded to further develop this technology by investigating the performance of larger scale versions of the design.
长期空间任务需要改进用于空间推进和存储系统的低温流体存储和传输技术。长期以来,筛选通道液体采集装置(LADs)一直与可储存推进剂一起使用,在发动机重启和液体传递过程中提供无蒸汽液体。由于低温流体的温度较低,在低温流体中使用lad是有问题的。外部热泄漏会导致在lad内形成蒸汽泡,在现有设计中难以去除。已经提出了一个锥形的LAD通道,可以可靠地去除设备内的蒸汽气泡,而无需昂贵的推力操作或主动分离系统。开发了一个模型来预测锥形LAD通道内的气泡运动,随后使用模拟流体完成了地面测试,以提供模型验证数据。锥形LAD技术的亚轨道微重力测试最近在两种不同的模拟流体中完成,并证明该概念可以被动地排出通道内的蒸汽泡。另外两次亚轨道飞行已经获得资助,通过研究该设计的更大规模版本的性能来进一步发展这项技术。
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引用次数: 0
The Circadian-clock Regulates the Arabidopsis Gravitropic Response 生物钟调节拟南芥向地性反应
J. Tolsma, Kaetlyn T. Ryan, Jacob J. Torres, J. Richards, Z. Richardson, Eric S. Land, I. Perera, Colleen J. Doherty
Abstract For long-term space missions, it is necessary to understand how organisms respond to changes in gravity. Plant roots are positively gravitropic; the primary root grows parallel to gravity's pull even after being turned away from the direction of gravity. We examined if this gravitropic response varies depending on the time of day reorientation occurs. When plants were reoriented in relation to the gravity vector or placed in simulated microgravity, the magnitude of the root gravitropic response varied depending on the time of day the initial change in gravity occurred. The response was greatest when plants were reoriented at dusk, just before a period of rapid growth, and were minimal just before dawn as the plants entered a period of reduced root growth. We found that this variation in the magnitude of the gravitropic response persisted in constant light (CL) suggesting the variation is circadian-regulated. Gravitropic responses were disrupted in plants with disrupted circadian clocks, including plants overexpressing Circadian-clock Associated 1 (CCA1) and elf3-2, in the reorientation assay and on a 2D clinostat. These findings indicate that circadian-regulated pathways modulate the gravitropic responses, thus, highlighting the importance of considering and recording the time of day gravitropic experiments are performed.
对于长期太空任务,有必要了解生物如何对重力变化做出反应。植物的根正向地性;主根即使在偏离重力方向后,也会与重力平行生长。我们研究了这种向地性反应是否会随着一天中重新定位发生的时间而变化。当植物相对于重力矢量重新定向或置于模拟微重力环境时,根系向地性响应的大小随重力初始变化发生的时间而变化。当植物在黄昏时重新定向时,就在快速生长期之前,这种反应是最大的,而在黎明前,当植物进入根系生长减少的时期时,这种反应最小。我们发现,这种向地性响应的大小变化在恒光(CL)下持续存在,这表明这种变化是受昼夜节律调节的。生物钟被打乱的植物,包括过度表达生物钟相关1 (CCA1)和elf3-2的植物,在定向实验和2D恒温器上的向地性反应被打乱。这些发现表明,昼夜调节的通路调节着向地性反应,因此,强调了考虑和记录一天中的向地性实验的重要性。
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引用次数: 3
The Adhesive Response of Regolith to Low-Energy Disturbances in Microgravity 微重力条件下风化层对低能扰动的黏附响应
S. Jarmak, J. Colwell, A. Dove, J. Brisset
Abstract Small, airless bodies are covered by a layer of regolith composed of particles ranging from μm-size dust to cm-size pebbles that evolve under conditions very different than those on Earth. Flight-based microgravity experiments investigating low-velocity collisions of cm-size projectiles into regolith have revealed that certain impact events result in a mass transfer from the target regolith onto the surface of the projectile. The key parameters that produce these events need to be characterized to understand the mechanical behavior of granular media, which is composed of the surfaces of small bodies. We carried out flight and ground-based research campaigns designed to investigate these mass transfer events. The goals of our experimental campaigns were (1) to identify projectile energy thresholds that influence mass transfer outcomes in low-energy collision events between cm-size projectiles and μm-size regolith, (2) to determine whether these mass transfer events required a microgravity environment to be observed, and (3) to determine whether the rebound portion of these collision events could be replicated in a laboratory drop tower environment. We found that (1) mass transfer events occurred for projectile rebound accelerations <7.8 m/s2 and we were unable to identify a corresponding impact velocity threshold, (2) mass transfer events require a microgravity environment, and (3) ourdrop tower experiments were able to produce mass transfer events. However, drop tower experiments do not exactly reproduce the free-particle impacts and rebound of the long-duration microgravity experiments and yielded systematically lower amounts of the overall mass transferred.
小的、没有空气的物体被一层风化层覆盖,这些风化层由μm大小的灰尘到cm大小的鹅卵石组成,它们在与地球上非常不同的条件下进化。基于飞行的微重力实验研究了厘米大小的弹丸与风化层的低速碰撞,揭示了某些撞击事件会导致质量从目标风化层转移到弹丸表面。产生这些事件的关键参数需要被表征,以理解颗粒介质的力学行为,它是由小物体的表面组成的。我们开展了旨在调查这些传质事件的飞行和地面研究活动。我们的实验活动的目标是:(1)确定影响cm大小弹丸与μm大小的风化层之间低能碰撞事件传质结果的弹丸能量阈值,(2)确定这些传质事件是否需要微重力环境才能观察到,(3)确定这些碰撞事件的反弹部分是否可以在实验室跌落塔环境中复制。我们发现(1)在弹射反弹加速度<7.8 m/s2时发生了传质事件,我们无法确定相应的冲击速度阈值;(2)传质事件需要微重力环境;(3)我们的跌落塔实验能够产生传质事件。然而,跌落塔实验并不能准确地再现长时间微重力实验的自由粒子撞击和反弹,并且系统地产生了较低的总传递质量。
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引用次数: 0
Reviewing Plasma Seed Treatments for Advancing Agriculture Applications on Earth and Into the Final Frontier 等离子体种子处理技术在推进农业在地球上的应用和进入最后前沿的综述
Annie Meier, Deborah Essumang, M. Hummerick, C. Johnson, M. Kruger, G. Massa, K. Engeling
Abstract With benefits such as environmentally safe treatment methods to stimulate growth, to increase plant yield, and improve disinfection efficiency, literature on the field of plasma treatment of seeds is growing. Generalized variables and success criteria have not been well correlated between studies, so this review paper serves to connect plasma and agriculture technologies to coordinate future efforts in this growing area of research. The authors have particular interest due to space agriculture, where seeds are sanitized before being sent into space for crop production. In order to supply a spectrum of nutritional needs, it is necessary to provide a variety of crops and ensure biological decontamination before the seeds are being sent into space. Traditional seed sanitization methods are not viable for all seed types, so exploration of other options is needed to expand the astronaut diet on long-duration space missions. This review paper brings together the current state-of-the-art reported literature to aide in understanding plasma seed application apparatus, seed or crop performance pertaining to germination, growth, water interactions, inactivation of bacteria, and surface sanitization results. These recent works include evolving research themes for potential seed treatment sanitization processes for various seed types to ensure the viability of plants for future growth in microgravity crop production systems.
摘要由于等离子体处理种子具有刺激生长、提高植物产量、提高消毒效率等环境安全的优点,有关等离子体处理种子领域的文献越来越多。广义变量和成功标准在研究之间没有很好的相关性,因此本文旨在将等离子体和农业技术联系起来,以协调这一不断发展的研究领域的未来努力。作者对太空农业特别感兴趣,因为在太空农业中,种子在被送入太空用于作物生产之前要经过消毒。为了满足一系列的营养需求,有必要提供各种各样的作物,并确保在种子被送往太空之前进行生物净化。传统的种子消毒方法并不适用于所有类型的种子,因此需要探索其他选择,以扩大长期太空任务中宇航员的饮食。这篇综述论文汇集了目前最先进的报道文献,以帮助理解等离子体施种装置,种子或作物的发芽,生长,水相互作用,细菌灭活和表面消毒结果。这些最近的工作包括不断发展的研究主题,包括各种种子类型的潜在种子处理消毒过程,以确保植物在微重力作物生产系统中未来生长的可行性。
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引用次数: 1
Pioneering the Approach to Understand a Trash-to-Gas Experiment in a Microgravity Environment 开创了理解微重力环境下垃圾变气体实验的方法
A. Meier, D. Rinderknecht, Joel A. Olson, M. Shah, Jaime A. Toro Medina, R. Pitts, R. Carro, J. Gleeson, J. Hochstadt, Evan A. Bell, Emily A. Forrester, M. Kruger, Deborah Essumang
Abstract The Orbital Syngas/Commodity Augmentation Reactor (OSCAR) project investigated hardware and engineering development for waste conversion operations related to trash deconstruction and repurposing for long duration space missions. Operations of the trash-to-gas system were investigated to compare microgravity (μg) and Earth gravity environments. The OSCAR system has been demonstrated in other μg platforms, but here the performance and results on the Blue Origin New Shepard Suborbital Vehicle are discussed. The OSCAR suborbital operation demonstrated the introduction of trash into a high temperature reactor for solid to gas conversion, ignition of mixed trash feedstock, combustion during μg, and subsequent gas collection processes in a flight automated sequence. An oxygen (O2)- and steam-rich environment was created within the reactor for ignition conditions, and the product gases were quantified to verify the reaction product composition. This paper focuses on the chemistry processes of the reactor, and gas and solid product analysis of the μg and gravity conditions. The gas production, reactor thermal profile, and mass and carbon conversion results validated confidence in the system design to continue the advancement of this technology for future spaceflight implementations.
轨道合成气/商品增强反应堆(OSCAR)项目研究了与垃圾解构和长时间太空任务重新利用相关的废物转化操作的硬件和工程开发。研究了垃圾制气系统的运行情况,比较了微重力环境和地球重力环境。OSCAR系统已经在其他平台上进行了演示,但这里讨论的是在蓝色起源新谢泼德亚轨道飞行器上的性能和结果。OSCAR亚轨道运行演示了将垃圾引入高温反应器进行固气转化、混合垃圾原料点火、μg过程中的燃烧以及随后在飞行自动化序列中的气体收集过程。在反应器内创造了一个富氧和富蒸汽的点火环境,并对产物气体进行了量化,以验证反应产物的组成。本文重点介绍了反应器的化学过程,以及气固产物的μg和重力条件分析。气体产量、反应堆热剖面、质量和碳转化结果验证了系统设计的信心,可以继续推进该技术在未来太空飞行中的应用。
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引用次数: 0
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