SEARCH · Engineering Papers

Results for “repair”

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

SourceResearch connectionFrom yearThrough year

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,171 records · Page 65

All sources NASA NTRS DOE OSTI

NASA NTRSDate not supplied

Ideas For Infusing In-Space Servicing, Assembly and Manufacturing Concepts into Nuclear Electric Propulsion Architectures

NASA is currently investigating nuclear electric propulsion (NEP) for human Mars transport within the space nuclear propulsion portfolio. NEP spacecraft have the following characteristics, they: 1) include very large structures (~100-meter length); 2) are comprised of many components/modules; and 3) have very long lifetimes (e.g., 50 years for fuel rods). Thus, NEP spacecraft can be classified as a “persistent asset,” which is any zero-g or planetary surface system that benefits from in-space assembly (ISA) or multiple visits for servicing, repairs, and upgrades. NEP spacecraft will benefit from taking advantage of, and incorporating, In-space Servicing, Assembly, and Manufacturing (ISAM) capabilities in the spacecraft architecture from the onset, enabling system maintenance, repair, and evolution. ISA has a long history of being proposed for, and studied as, a means for achieving large systems in space. More recently, the benefits of ISA have been recognized by NASA, the Department of Defense (DOD), other government agencies, and commercial space companies, and thus, ISAM is being actively pursued at a national level. Past and current strategies for achieving large structures in space have relied largely on two strategies; the first is to launch monolithic structures (designed to meet launch vehicle requirements for payload size and mass) that are docked or berthed to other monolithic structures on-orbit to form a larger structure (e.g., the International Space Station [ISS]); the second is folding and packaging large structures to fit inside a payload fairing and deploying the full-sized structure (unaided) once on-orbit (e.g., the James Webb Space Telescope [JWST]). To date, conceptual architecture studies performed for NEP spacecraft capable of human-rated Mars transport have only included a combination of the two previously mentioned strategies. This paper will propose ideas for infusing ISAM strategies into NEP vehicle architectures that leverage existing and near future technologies and enable the resulting NEP systems to be realized in a more time- and cost-efficient manner.

in-space assembly↗

NASA NTRSDate not supplied

Ideas for Infusing In-Space Servicing, Assembly, and Manufacturing Concepts into Nuclear Electric Propulsion Architectures

in-space assembly↗

NASA NTRSDate not supplied

Spaceflight Autonomous Multigenerational Microbial Sequencer in Support of Plant-Growth Systems

The CubeSat platform has proven successful in obtaining meaningful life science information when biological payloads are incorporated. Examples include: 1) the first-ever CubeSat with a biological payload, GeneSat-1, which demonstrated decreased growth rates for flight samples of Escherichia coli in low Earth orbit (Parra et al. 2008); 2) PharmaSat, demonstrated that Saccharomyces cerevisiae in the microgravity environment exhibits a significant level of metabolic activity even at high doses of applied antifungal (Ricco et al. 2011); 3) O/OREOS, which used Bacillus subtilis(bacteria) to demonstrate for the first time that microorganisms can be loaded in a dried, dormant form and then rehydrated and grown in orbit months after launch (Nicholson et al. 2011; Ehrenfreund et al. 2014; 4) the SporeSat payload, which investigated Ceratopteris richardii(fern spores) using lab-on-a-chip devices (BioCDs) and minicentrifuges to produce artificial gravitational forces in ground studies (Park et al. 2017), with demonstration of the BioCD and minicentrifuge in space; 5) EcAMSat, the first CubeSat to be directly deployed from the ISS for an experiment assessing antibiotic resistance of E. coli in the microgravity environment (Padgen et al. 2020); 6) BioSentinel, exposed a culture of yeast to galactic cosmic radiation (GCR) and solar particle events while in heliocentric orbit to measure the rate of double-strand-break repair using DNA-repair-deficient mutants. This effort measures the metabolic parameters of yeast in a deep-space environment compared to Earth ambient conditions using a 3-color LED detection system (Ricco et al. 2020; Padgen et al. 2021). We aim to expand this list to include a Spaceflight Autonomous Multigenerational Microbial Sequencer (SAMMS). SAMMS will allow for the genome level understanding of changes in growth and metabolic activity for any organism. While microbes are suitable for early studies in our proposed platform because of their small size, small and relatively less-complicated genomes, fast generation times, and relevance to life support systems; multicellular organisms can similarly be evaluated for their genetic response to the spaceflight environment. The Spaceflight Autonomous Multigenerational Microbial Sequencer (SAMMS) will enable autonomous sequencing of biological samples in plant production units, cislunar orbit and on the lunar surface to examine spaceflight effects (ie. radiation, altered gravity, reduced pressures) on plant and microbial genomes.On this team a Kennedy Space Center (KSC) space crop production and water systems microbiologist/molecular biologist works with a Johnson Space Center (JSC) International Space Station (ISS) microbial sequencing expert and an Ames Research Center (ARC) CubeSat Engineering team to convert an automated Oxford Nanopore librarypreparation and sequencing method to a fluidic CubeSat payload system. The Oxford Nanopore MinION sequencing platform has proven successful in the spaceflight environment onboard the ISS (Stahl-Rommel et al. 2021). Further long-duration spaceflight and exposure to high levels of radiation will cause genotypic effects in biological organisms that may affect their function. Monitoring the adaption of a population to the spaceflight environment and any subsequent beneficial mutations will allow for the harnessing of organisms best suited for use in life support systems. This will ensure that the selected life support-essential microorganisms maintain their intended specified function over generations of culturing in the relevant spaceflight environment without becoming hazardous to crew or spacecraft systems.

Aubrie E Orourke↗

NASA NTRSDate not supplied

Automated Reconfigurable Mission Adaptive Digital Assembly Systems (ARMADAS): Robotically Assembled Sustainable Lunar Infrastructure

Introduction: The Automated Reconfigurable Mission Adaptive Digital Assembly Systems (ARMADAS) project at NASA Ames Research Center is developing autonomous infrastructure, instrumentation, and spacecraft assembly and manufacturing capabilities for next generation exploration and science missions, with a goal to change the cost scaling of these missions relative to mission size and duration. Using a building-block approach with a 'kit of parts' composed of ultra-light, high-performance mechanical metamaterials, simplified robots leverage the period environment to achieve high levels of autonomy and reliability for in-space and surface assembly of large-scale apertures, solar-arrays, towers, habitats, and other infrastructure. Robots and structure break down into a compact form factor for launch. By leveraging economies of scale and achieving high-packing ratios, ARMADAS technology can revolutionize possible space missions by breaking the tyranny of the launch shroud, decreasing development times, decreasing mission costs for transformative science capability, and providing a scalable and versatile space infrastructure strategy. Capabilities: To date, the ARMADAS project has demonstrated autonomous assembly of large numbers of structural modules into a meters-scale structure in an earth gravity environment. The mechanical performance is on par with conventional space structures, at a fraction of the cost. By using highly repeatable manufacturing processes (injection molding carbon fiber reinforced space-rated polymers), the structures are both cost effective and highly precise. The ARAMADAS system carefully designs parts for high-precision assembly–the simple and cost-effective robots build structures much larger and more precise than themselves. While maintaining structural efficiency, the ARMADAS system encompasses many functional module types, including for solar power, comm/power routing, etc., that will enable entire infrastructure systems to be engineered and autonomously assembled with modular parts. With well-defined interfaces, custom instrumentation modules can easily be added to fit many mission objectives. Lunar Infrastructure Vision: At the LSIC spring meeting, ARMADAS will present a vision for a general-purpose lunar construction kit capable of meeting a wide variety of lunar surface infrastructure needs (Figure 1). Investing in an ecosystem of reconfigurable, discretely repairable parts and robots enables systems than can expand their capability, reconfigure to meet emergency or unforeseen needs, self-repair and reduce spare-part needs. With a small set of parts (module types), a wide variety of infrastructure needs can be met. Footing modules will allow infrastructure construction at locations with no surface preparation. Footer, primary structure, and rail modules can create reconfigurable rail systems. These rail systems can be used to reduce cost-of-transport between frequently accessed sites, provide dust mitigation, and convey power and communications. High-performance structure modules combined with power-routing and solar modules can create tall towers for power and communication. Concepts for habitats and garages encompass a structure to support regolith cover for a pre-integrated module to an entirely ARMADAS system-based structure. This system can be leveraged by many in-situ resource utilization (ISRU) technologies to simplify processes and augment capabilities.

C. E. Gregg↗

NASA NTRSDate not supplied

Engineered Yeast to Test Risks for Human Exploration of the Lunar Surface

Jessica W. Chau, Natalie N. Ball, Aditya Hindupur, Sandra T. Vu, Jennifer Gil Acevedo, Lauren C. Liddell, Chinmayee Govinda Raj, Gentry, Sergio R. Santa Maria, A. Mark Settles Crewed exploration of the Moon carries risks of long duration exposure to reduced gravity and to deep space radiation. The Lunar Explorer Instrument for space biology Applications (LEIA) investigates the effects of increased radiation and reduced gravity on yeast viability and growth in a Commercial Lunar Payload Services (CLPS) surface mission to the south polar region. LEIA conducts yeast genetics experiments to quantify growth, metabolism, and synthetic biology-enabled production of human nutrients, while taking real time measurements of biologically relevant radiation exposure on the lunar surface. We have engineered beta-carotene producing yeast strains to test the importance of selected DNA damage repair and reactive oxygen species (ROS) defense pathways in mitigating cellular damage from lunar surface radiation. Carotenoids are important dietary antioxidants, and beta-carotene is pro-vitamin A, which is needed for vision and immune function. Carotenoids are sensitive to ROS produced by ionizing radiation and NASA is testing on-demand production of carotenoids from yeast in the BioNutrients space flight experiments. In LEIA, we test the effects of deep space on carotenoid yield in engineered yeast strains. The LEIA team uses CRISPR-Cas9 to engineer yeast to express carotenoids as well as to generate loss-of-function mutations. We are generating mutations in the RAD51 DNA damage repair locus and three genes that function to reduce oxidative damage to the cell: SOD1, SOD2, and TSA1. These strains are tested for carotenoid production using microfluidics and LED spectroscopy to allow remote sensing of cellular growth and carotenoid levels. Keywords: synthetic biology, oxidative stress tolerance, biosensors, space radiation, beyond low Earth orbit, lunar surface, CRISPR/Cas9, gene editing, desiccation, carotenoids.

synthetic biology↗

NASA NTRSDate not supplied

Gene Expression of Peripheral Blood Mononuclear Cells of Crew Members During Long-Duration Space Missions Indicate Dysregulation of Immunological and Cell Survival Mechanisms

Lymphocytes are naturally exposed to genotoxic stresses. DNA damage occurs during the entire lymphocyte’s life span and is induced mainly by reactive oxygen species (ROS), replication fork collapse, or telomere shortening during the immune response or intense cell proliferation phases. Strong evidence for the influence of immune function on DNA repair comes from studies of SCID disease. SCID mice not only have a deficient V(D)J recombination but are also unable to repair double strand breaks, leading to increased radiation sensitivity. The leukocytes’ transcriptome of 8 ISS crew members revels a dysregulated immune function and activation of cellular survival pathways in response to space environment. We have performed PCR analysis in peripheral mononuclear cells from the same crew members. A list of 62 genes were carefully selected addressing immunological and cell survival pathways. Differentially expressed genes indicated changes in chemokine receptor activity, chemokine binding, toll-like receptors, adhesion molecules and cellular response to DNA damage.

Maria Moreno-Villanueva↗

NASA NTRS2024

Gene Expression of Peripheral Blood Mononuclear Cells of Crew Members During Long-Duration Space Missions Indicate Dysregulation of Immunological and Cell Survival Mechanisms

María Moreno-Villanueva↗

NASA NTRSDate not supplied

Evaluation of the KOKEE12M/ONSA13NE Baseline as a KOKEE12M/WETTZ13S Backup

USNO's IERS Rapid Service/Prediction Center (RS/PC) uses the KOKEE12M/WETTZ13S (K2/Ws) baseline for its combination solutions. It would be useful for the RS/PC to be able to use backup baselines in case K2 or Ws is not available. While Ws was undergoing repair in late 2022 through early 2023, ONSA13NE (Oe) replaced Ws to provide immediate UT1 for IVS Analysis Center products but also to begin building a data set for the RS/PC to eventually use to ``characterize'' (assess and approve) the K2/Oe baseline for use in its solutions. The RS/PC needs 60 sessions to be able to characterize a baseline, and the data from the repair period provided 58 sessions, with three additional sessions being observed later. Here we evaluate the K2/Oe baseline to see if it is a viable candidate for submission to the RS/PC for characterization. We evaluate the K2/Oe baseline itself and also compare the K2/Oe baseline to the K2/Ws baseline.

VLBI↗

NASA NTRSDate not supplied

Modular Interface for CLPS-Scale Excavators (MICE) – Feasibility Testing of a Modular Disconnect System for Regolith Manipulation Implements Focused on Excavation and Site Preparation Activies

The Artemis Program seeks to establish a sustained lunar presence and robust economy driven by re-sources available on the Moon and enabled by sup-porting infrastructure. This will require robotic manipulation of tens of thousands of metric tons of regolith. Excavation and site preparation tasks in the unforgiving lunar environment will inevitably lead to component wear and necessitate periodic maintenance and repair. System modularity and standard interfaces are key components of enabling robotic repair and maintenance. The Modular Interface for CLPS (Commercial Lunar Payloads Services) -scale Excavators (MICE) project is a funded fiscal year 2024 Internal Research and Development (IRAD) project at NASA KSC. The MICE project seeks to test concepts for modular re-configuration of a CLPS-scale excavation and site preparation rover. These concepts, henceforth referred to as Modular Disconnect Systems (MDS), allow a mobility platform to interface with regolith manipulation implements such as a bucket drum excavator or compactor. The MDS concepts allow the mobility platform to detect one implement and swap to another implement in-situ. Power and communications is fed through a dust-tolerant electrical connection on the MDS to the implement’s actuator(s) and sensors.

MICE↗

NASA NTRS2024

Wilson Corners, Solid Waste Management Unit 001(SWMU 01) 2023 Annual Long-Term Monitoring Report

This report presents a summary of the long-term monitoring (LTM) activities that occurred in 2023 at Wilson Corners, Solid Waste Management Unit 001, at Kennedy Space Center (KSC), Florida. Annual LTM of groundwater is being conducted at the site. Based on results from groundwater sampling activities performed during the 2019 through 2020 LTM reporting period and the 2020 and 2021 DPT groundwater sampling, it was determined that the LTM sampling plan was no longer meeting the goal of LTM because delineation was not verified and the installation of an air sparge (AS) system to treat the area of the High Concentration Plume was recommended. The AS System was installed in late 2022 and early 2023. System start-up activities were initiated in April 2023. Following system startup, several site wells required retrofitting to equip wellheads for withstanding the air pressure released from air sparge wells during system operation. Some site wells also required repair or abandonment, and replacement. Survey of location and top-of-casing of newly installed monitoring wells was combined with scheduled AS system survey activities and was completed in January 2024. The activities presented in this report include the February and April 2023 LTM monitoring well installations; March and April 2023 LTM and performance monitoring well water level gauging and sampling; November 2023 LTM well retrofits and repairs; a summary of December 2023 LTM well abandonments and installations (complete site well abandonment activities will be presented under a separate cover); and January 2024 LTM well survey. During the March and April 2023 sampling events, the low-flow sampling method was used, and samples were analyzed for a select list of volatile organic compounds. In March 2023, groundwater flow for the site was generally to the west was generally consistent with historical observations at the site. The Low Concentration Plume (LCP) continues to extend both horizontally and vertically beyond the terminal depth of the current monitoring well network. Data, inclusive of the 2023 LTM and baseline performance monitoring sampling events, indicate that the LCP encompasses an estimated 19.5 acres, compared to the 2021 LCP footprint, inclusive of the 2020 and 2021 DPT sampling events of 20.7 acres. The vertical extent of VOCs was historically delineated by monitoring wells screened greater than 48 feet below land surface (bls). The results from the three vertical extent monitoring wells screened below 48 feet bls that were sampled during the 2023 LTM indicate that groundwater vinyl chloride concentrations in these three wells are greater than the GCTL. As presented in the 2021 Long-Term Monitoring Report (NASA 2022), the KSCRT agreed to delay deeper investigations in this area to prevent the creation of additional pathways for vertical migration. Based on groundwater sampling activities performed in 2023, recommendations are to perform the next annual LTM sampling event, scheduled for April 2024 and to conduct quarterly performance monitoring of the AS System. The current selection of monitoring wells in the recommended 2024 LTM plan will provide an adequate data set for monitoring groundwater plume behavior; however, the LTM monitoring well network will be evaluated and refined based on 2024 LTM and year one performance monitoring data.

King Linnea↗

NASA NTRSDate not supplied

Computational and Physics-Based Modeling for the Development of in-Space Welding Technology

Welding will be an enabling technology for in-space assembly and manufacturing (ISAM) and in-space repair operations in support of the growing Space Economy. Prospective use cases of welding include metal repair on long-duration lunar surface missions and assembly of large structures that exceed dimensions of the currently available launch vehicle envelopes such as space arrays, tall lunar transmission towers, or even large transit vehicles powered by nuclear propulsion. There is a brief history of beam welding for in-space use. Soviet and NASA astronauts performed electron beam welds (EBW) in space in the late-1960s through mid-1980s and several parabolic flights of laser beam welding (LBW) were undertaken in the USA and Canada in the early-1990s. Work has since stagnated until the recent boom of the in-space economy has demonstrated need for continued development of EBW and LBW processes, which are desirable for their compactness, low mass, low power consumption, and high wall plug efficiency. No space-based manufacturing laboratory capable of demonstrating, developing, and applying welding processes exists, and costs are high to develop and fly individual space-based trials. This drives the current approach to develop welding technology with computational and physics-based models in tandem with earth-based experiments. Several approaches are taken to complement and drive the selection of instrumentation for planned parabolic and suborbital flights. First, a thermal model of Skylab EBW experiments is established and compared to empirical data. Next, the model is used to show variation between Earth-based and in-space weld structure and properties through exploration of key variables that define the Low earth orbit and lunar surface space environments, namely temperature (40 – 400 K), pressure (10-10 to 104 Pa), and gravity (10-6 g to 1 g). Finally, physical processes associated with heat and mass transport during welding are evaluated through several case studies across the above variable ranges demonstrating need for further research.

in-space welding↗

DOE OSTI2023

SyReNN: A tool for analyzing deep neural networks

Deep Neural Networks (DNNs) are rapidly gaining popularity in a variety of important domains. Unfortunately, modern DNNs have been shown to be vulnerable to a variety of attacks and buggy behavior. This has motivated recent work in formally analyzing the properties of such DNNs. Furthermore, this paper introduces SyReNN, a tool for understanding and analyzing a DNN by computing its symbolic representation. The key insight is to decompose the DNN into linear functions. Our tool is designed for analyses using low-dimensional subsets of the input space, a unique design point in the space of DNN analysis tools. We describe the tool and the underlying theory, then evaluate its use and performance on three case studies: computing Integrated Gradients, visualizing a DNN’s decision boundaries, and repairing buggy DNNs.

97 MATHEMATICS AND COMPUTING↗

DOE OSTI2021

Positron Emission Tomography Imaging of Functional Transforming Growth Factor β (TGFβ) Activity and Benefit of TGFβ Inhibition in Irradiated Intracranial Tumors

Transforming growth factor β (TGFβ) promotes cell survival by endorsing DNA damage repair and mediates an immunosuppressive tumor microenvironment. Thus, TGFβ activation in response to radiation therapy is potentially targetable because it opposes therapeutic control. Strategies to assess this potential in the clinic are needed.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

DOE OSTI2023

Functional analysis of a conserved site mutation in the DNA end processing enzyme PNKP leading to ataxia with oculomotor apraxia type 4 in humans

Polynucleotide kinase 3'-phosphatase (PNKP), an essential DNA end-processing enzyme in mammals with 3'-phosphatase and 5'-kinase activities, plays a pivotal role in multiple DNA repair pathways. Its functional deficiency has been etiologically linked to various neurological disorders. Recent reports have shown that mutation at a conserved glutamine (Gln) in PNKP leads to late-onset ataxia with oculomotor apraxia type 4 (AOA4) in humans and embryonic lethality in pigs. However, the molecular mechanism underlying such phenotypes remains elusive. Here, we report that the enzymatic activities of the mutant versus WT PNKP are comparable; however, cells expressing mutant PNKP and peripheral blood mononuclear cells (PBMCs) of AOA4 patients showed a significant amount of DNA double-strand break accumulation and consequent activation of the DNA damage response. Further investigation revealed that the nuclear localization of mutant PNKP is severely abrogated, and the mutant proteins remain primarily in the cytoplasm. Western blot analysis of AOA4 patient-derived PBMCs also revealed the presence of mutated PNKP predominantly in the cytoplasm. To understand the molecular determinants, we identified that mutation at a conserved Gln residue impedes the interaction of PNKP with importin alpha but not with importin beta, two highly conserved proteins that mediate the import of proteins from the cytoplasm into the nucleus. Collectively, our data suggest that the absence of PNKP in the nucleus leads to constant activation of the DNA damage response due to persistent accumulation of double-strand breaks in the mutant cells, triggering death of vulnerable brain cells—a potential cause of neurodegeneration in AOA4 patients.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2020

Uncovering DNA-PKcs ancient phylogeny, unique sequence motifs and insights for human disease

DNA-dependent protein kinase catalytic subunit (DNA-PKcs) is a key member of the phosphatidylinositol-3 kinase-like (PIKK) family of protein kinases with critical roles in DNA-double strand break repair, transcription, metastasis, mitosis, RNA processing, and innate and adaptive immunity. The absence of DNA-PKcs from many model organisms has led to the assumption that DNA-PKcs is a vertebrate-specific PIKK. Here, we find that DNA-PKcs is widely distributed in invertebrates, fungi, plants, and protists, and that threonines 2609, 2638, and 2647 of the ABCDE cluster of phosphorylation sites are highly conserved amongst most Eukaryotes. Furthermore, we identify highly conserved amino acid sequence motifs and domains that are characteristic of DNA-PKcs relative to other PIKKs. These include residues in the Forehead domain and a novel motif we have termed YRPD, located in an α helix C-terminal to the ABCDE phosphorylation site loop. Combining sequence with biochemistry plus structural data on human DNA-PKcs unveils conserved sequence and conformational features with functional insights and implications. The defined generally progressive DNA-PKcs sequence diversification uncovers conserved functionality supported by Evolutionary Trace analysis, suggesting that for many organisms both functional sites and evolutionary pressures remain identical due to fundamental cell biology. The mining of cancer genomic data and germline mutations causing human inherited disease reveal that robust DNA-PKcs activity in tumors is detrimental to patient survival, whereas germline mutations compromising function are linked to severe immunodeficiency and neuronal degeneration. We anticipate that these collective results will enable ongoing DNA-PKcs functional analyses with biological and medical implications.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2021

Synaptonemal complex formation produces a particular arrangement of the lateral element-associated DNA

Highlights: • The NAMA-Ur method can be used to analyze the DNA associated to the synaptonemal complex. • The DNA associated to the synaptonemal complex display bubble-like organization. • Formation of the synaptonemal complex is enough to produce the bubble-like organization of the DNA. • Synaptonemal complex without lateral elements produce similar organization of its associated DNA. During meiosis, homologous chromosomes exchange genetic material. This exchange or meiotic recombination is mediated by a proteinaceous scaffold known as the Synaptonemal complex (SC). Any defects in its formation produce failures in meiotic recombination, chromosome segregation and meiosis completion. It has been proposed that DNA repair events that will be resolved by crossover between homologous chromosomes are predetermined by the SC. Hence, structural analysis of the organization of the DNA in the SC could shed light on the process of crossover interference. In this work, we employed an ultrastructural DNA staining technique on mouse testis and followed nuclei of pachytene cells. We observed structures organized similarly to the SCs stained with conventional techniques. These structures, presumably the DNA in the SCs, are delineating the edges of both lateral elements and no staining was observed between them. DNA in the LEs resembles two parallel tracks. However, a bubble-like staining pattern in certain regions of the SC was observed. Furthermore, this staining pattern is found in SCs formed between non-homologous chromosomes, in SCs formed between sister chromatids and in SCs without lateral elements, suggesting that this particular organization of the DNA is determined by the synapsis of the chromosomes despite their lack of homology or the presence of partially formed SCs.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2021

Long noncoding RNA MEG3 expressed in human dental pulp regulates LPS-Induced inflammation and odontogenic differentiation in pulpitis

Highlights: • LncRNA MEG3, upregulated in inflamed dental pulp, is involved in the inflammation and regeneration in pulpitis. • The expression of lncRNA MEG3 was semiquantitatively detected in the pulp tissue by the innovative RNAscope® technique. • MEG3 downregulation can inhibit inflammatory cytokines secretion and also promote the odontogenic differentiation of hDPCs. • LncRNA MEG3 can be a potential therapeutic target in the inflammation and regeneration of dentin-pulp complex in pulpitis. Pulpitis refers to inflammation of the inner pulp by invading microbes, and tissue repair occurs due to odontogenic differentiation of human dental pulp cells (hDPCs) with multidifferentiation potential. Long noncoding RNAs (lncRNAs) can modulate numerous pathological and biological processes; however, the role of lncRNAs in the inflammation and regeneration of the dentin-pulp complex in pulpitis is unclear. Here, we performed high-throughput sequencing to identify differentially expressed lncRNAs between human normal and inflamed pulp and concluded that lncMEG3 (lncRNA maternally expressed gene 3, MEG3) was signicantly upregulated in both inflamed pulp and LPS-treated hDPCs. MEG3 expression in the pulp tissue was detected using the RNAscope® technique. RNA pulldown assays identified the MEG3-interacting proteins and the potential mechanisms. With MEG3 knockdown, we investigated the role of MEG3 in the secretion of inflammatory cytokines in LPS-treated hDPCs and odontogenic differentiation of hDPCs. MEG3 downregulation inhibited the secretion of TNF-α, IL-1β and IL-6 in LPS-treated hDPCs, and the p38/MAPK signaling pathway may be related to this effect. MEG3 knockdown promoted odontogenic differentiation of hDPCs by regulating the Wnt/β-catenin signaling pathway. Our study suggested that MEG3 has a negative effect on inflammation and regeneration of the dentin-pulp complex in pulpitis.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2021

Dual role of Fam208a during zygotic cleavage and early embryonic development

Maintenance of genome stability is essential for every living cell as genetic information is repeatedly challenged during DNA replication in each cell division event. Errors, defects, delays, and mistakes that arise during mitosis or meiosis lead to an activation of DNA repair processes and in case of their failure, programmed cell death, i.e. apoptosis, could be initiated. Fam208a is a protein whose importance in heterochromatin maintenance has been described recently. In this work, we describe the crucial role of Fam208a in sustaining genome stability during cellular division. }The targeted depletion of Fam208a in mice using CRISPR/Cas9 led to embryonic lethality before E12.5. We also used the siRNA approach to downregulate Fam208a in zygotes to avoid the influence of maternal RNA in the early stages of development. This early downregulation increased arresting of the embryonal development at the two-cell stage and the occurrence of multipolar spindles formation. To investigate this further, we used the yeast two-hybrid (Y2H) system and identified new putative interaction partners Gpsm2, Svil, and Itgb3bp. Their co-expression with Fam208a was assessed by RT-qPCR profiling and in situ hybridization [1] in multiple murine tissues. Based on these results we proposed that Fam208a functions within the HUSH complex by interaction with Mphosph8 as these proteins are not only able to physically interact but also co-localise. We are bringing new evidence that Fam208a is a multi-interacting protein affecting genome stability on the cell division level at the earliest stages of development and by interaction with methylation complex in adult tissues. }In addition to its epigenetic functions, Fam208a appears to have an important role in the zygotic division, possibly via interaction with newly identified putative partners Gpsm2, Svil, and Itgb3bp.

60 APPLIED LIFE SCIENCES↗