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At least 109 records · Page 6

Glucose-dependent insulinotropic peptide stimulates thymidine incorporation in endothelial cells: role of endothelin-1

We have previously characterized the receptor for glucose-dependent insulinotropic polypeptide (GIPR) in vascular endothelial cells (EC). Different EC types were found to contain distinct GIPR splice variants. To determine whether activation of the GIPR splice variants resulted in different cellular responses, we examined GIP effects on human umbilical vein endothelial cells (HUVEC), which contain two GIPR splice variants, and compared them with a spontaneously transformed human umbilical vein EC line, ECV 304, which contains four GIPR splice variants. GIP dose-dependently stimulated HUVEC and ECV 304 proliferation as measured by [3H]thymidine incorporation. GIP increased endothelin-1 (ET-1) secretion from HUVEC but not from ECV 304. Use of the endothelin B receptor blocker BQ-788 resulted in an inhibition of [3H]thymidine incorporation in HUVEC but not in ECV 304. These findings suggest that, although GIP increases [3H]thymidine incorporation in both HUVEC and ECV 304, this proliferative response is mediated by ET-1 only in HUVEC. These differences in cellular response to GIP may be related to differences in activation of GIPR splice variants.

NASA Discipline Clinical Medicine↗

Decoding co-/post-transcriptional complexities of plant transcriptomes and epitranscriptome using next-generation sequencing technologies

Next-generation sequencing (NGS) technologies - Illumina RNA-seq, Pacific Biosciences isoform sequencing (PacBio Iso-seq), and Oxford Nanopore direct RNA sequencing (DRS) - have revealed the complexity of plant transcriptomes and their regulation at the co-/post-transcriptional level. Global analysis of mature mRNAs, transcripts from nuclear run-on assays, and nascent chromatin-bound mRNAs using short as well as full-length and single-molecule DRS reads have uncovered potential roles of different forms of RNA polymerase II during the transcription process, and the extent of co-transcriptional pre-mRNA splicing and polyadenylation. These tools have also allowed mapping of transcriptome-wide start sites in cap-containing RNAs, poly(A) site choice, poly(A) tail length, and RNA base modifications. The emerging theme from recent studies is that reprogramming of gene expression in response to developmental cues and stresses at the co-/post-transcriptional level likely plays a crucial role in eliciting appropriate responses for optimal growth and plant survival under adverse conditions. Although the mechanisms by which developmental cues and different stresses regulate co-/post-transcriptional splicing are largely unknown, a few recent studies indicate that the external cues target spliceosomal and splicing regulatory proteins to modulate alternative splicing. In this review, we provide an overview of recent discoveries on the dynamics and complexities of plant transcriptomes, mechanistic insights into splicing regulation, and discuss critical gaps in co-/post-transcriptional research that need to be addressed using diverse genomic and biochemical approaches.

Biochemistry & Molecular Biology↗

Lunar Module Wiring Design Considerations and Failure Modes

This slide presentation reviews the considerations for the design of wiring for the Lunar Module. Included are a review of the choice of conductors and insulations, the wire splicing (i.e., crimping, and soldering), the wire connectors, and the fabrication of the wire harnesses. The problems in fabrication include the wires being the wrong length, the damage due to the sharp edges, the requried use of temproary protective covers and inadequate training. The problems in the wire harness installation include damge from sharp eges, work on adjacent harnesses, connector damage, and breaking wires. Engineering suggestions from the Apollo-era in reference to the conductors that are reviewed include: the use of plated conductors, and the use of alloys for stronger wiring. In refernce to insulation, the suggestions from Apollo era include the use of polymer tape-wrap wire insulation due to the light weight, however, other types of modern insulation might be more cost-effective. In reference to wire splices and terminal boards the suggestions from the Apollo Era include the use of crimp splices as superior to solder splices, joining multiple wire to a common point using modular plug-ins might be more reliable, but are heavier than crimp splicing. For connectors, the lessons from the Apollo era indicate that a rear environmental seal that does not require additional potting is preferred, and pins should be crimped or welded to the incoming wires and be removable from the rear of the connector.

Interbartolo, Michael↗

Hardware in the Loop Performance of Terrestrial Powered Descent Dual Quaternion Guidance With A Custom First-Order Solver

The Safe & Precise Landing Integrated Capabilities Evolution (SPLICE) program continues to push the development of advanced descent and landing technologies. This paper will focus on the improvements made to the SPLICE Dual Quaternion Guidance (DQG) algorithm, which is a critical software component for generating approach phase and hazard avoidance trajectories. The trajectory performance of SPLICE DQG will be evaluated in preparation for a terrestrial flight test that incorporates a hazard avoidance maneuver. Additionally, the implementation of a customized first order Proportional-Integral Projected Gradient solver will be reviewed, along with preliminary execution results on the SPLICE Descent and Landing Computer (DLC).

SPLICE↗

Technique for recovery of voice data from heat damaged magnetic tape

A method for conditioning, and thus enabling retrieval of intelligence from, magnetic tapes after damage from heat has caused the tape to wrinkle and curl severely thereby reducing tape width to less than one-half its original size. The damaged tape is superposed on a first piece of splicing tape with the oxide side of the magnetic tape in contact with the adhesive side of the splicing tape and then carefully smoothed by a special tool. A second piece of splicing tape is placed on the backing side of the magnetic tape then the resulting tape stack is trimmed to the original width of the magnetic tape. After the first piece of splicing tape is carefully removed from the oxide side of the damaged magnetic tape, the resulting magnetic tape is then ready to be placed into a recorder for playback.

Melugin, J. F.↗

Apollo 6, Anomaly Report No. 6: Abnormal Structural Performance During Launch Phase

Approximately 2 minutes 13 seconds after lift-off of the Apollo 6 mission, abrupt changes of strain, vibration, and acceleration measurements were indicated in the S-IVB, instrument unit, adapter, lunar module, and command and service modules; photographs showed objects coming from the area of the adapter. The adapter, however, continued to sustain the required loads with no impairment of the mission. The investigation was first focused upon the understanding of the coupled vibration modes and characteristics of the launch vehicle and spacecraft.' Extensive test programs were conducted. It was eventually concluded that the adapter failure was not caused by vibration. Extensive study of the airborne photography and other evidence indicated that a large area of the adapter had lost inner facesheet from the honeycomb sandwich panels. Loads and stresses resulting from vibration were determined to be insufficient to initiate such a failure. The investigation was then directed toward determining the range of pressures that could have been trapped in the Apollo 6 adapter sandwich panels, and toward determining the tolerance of the panels to withstand pressure with various degrees of flaws such as adhesive voids and facesheet dents. The degradation effects of moisture and heat exposure on the adhesive strength were also studied and tested. These tests and analyses led to the conclusion that pressure internal to the sandwich panels could have caused the failure, if a large flaw existed. The pressure buildup would have been caused by aerodynamic heating effects on air and moisture trapped in the panel. The probable cause of the failure was found in the original ultrasonic inspection scan record of the affected adapter panel. In the center of the region where the adapter failed, horizontally along the station 709 panel splice, the record contained two thick anomalous lines extending several feet. Without an X-ray record of this region, the significance of this particular scan record cannot be fully understood. However, since all other evidence had indicated that the adhesive had to be weakened in a rather large area to initiate the failure, the investigation was focused intently upon the station 709 splices of other adapters. Sufficient information was developed to verify that deficient assembly techniques have consistently resulted in abnormalities in the structure at this station. These abnormalities were identified in adapters 12, 13, 14, 15, and 16. Before the splice abnormalities were pinpointed, corrective action was taken to reduce pressure "buildup in the honeycomb panels and to reduce heat degrading effects on the adhesive. This was done by drilling vent holes in the inner facesheet and covering the outer facesheet with cork. The adapters having identified abnormalities in the station 709 splice are being repaired, and the contractor is investigating ways of avoiding these -abnormalities in panels yet to be bonded.

Source record↗

NASA and Blue Origin’s Flight Assessment of Precision Landing Algorithms Computing Performance

NASA’s Safe and Precise Landing - Integrated Capabilities Evolution (SPLICE) project continues NASA’s work in the development and testing of technologies for Precision Landing and Hazard Avoidance (PL&HA). This paper presents results characterizing how SPLICE flight software utilizes the shared computing resources of the Descent Landing Computer (DLC), one of the PL&HA technologies under development. The SPLICE technologies are being tested as an integrated payload on Blue Origin’s New Shephard suborbital vehicle. The results presented in this paper are measured by applications running in and with the flight software both in flight, and in a high-fidelity Hardware-in-the-Loop (HWIL) simulation environment. Linux utilities to measure performance are also executed from the command line in the HWIL configuration. Performance measurements of the SPLICE workloads executing on the DLC provide insight on how efficiently the software is utilizing the DLC resources. Examples of how these measurements have guided improvements in the flight code are presented. In addition, the DLC uses a commercial processor as a surrogate for NASA’s High Performance Spaceflight Computing (HPSC) processor. This work provides insight on how an HPSC system may perform delivering PL&HA capabilities on a future mission. The measurements also can be used to infer architectural requirements for PL&HA capabilities, informing the HPSC project and other flight computer development efforts. Examples of the measurements collected include processor utilization, I/O bandwidth, cache and branch misses, and application profiles.

Precision Landing and Hazard Avoidance↗

NASA and Blue Origin’s Flight Assessment of Precision Landing Algorithms Computing Performance

NASA’s Safe and Precise Landing - Integrated Capabilities Evolution (SPLICE) project continues NASA’s work in the development and testing of technologies for Precision Landing and Hazard Avoidance (PL&HA). This paper presents results characterizing how SPLICE flight software utilizes the shared computing resources of the Descent Landing Computer (DLC), one of the PL&HA technologies under development. The SPLICE technologies are being tested as an integrated payload on Blue Origin’s New Shepard suborbital vehicle. The results presented in this paper are measured by applications running in and with the flight software both in flight, and in a high-fidelity Hardware-in-the-Loop (HWIL) simulation environment. Linux utilities to measure performance are also executed from the command line in the HWIL configuration. Performance measurements of the SPLICE workloads executing on the DLC provide insight on how efficiently the software is utilizing the DLC resources. Examples of how these measurements have guided improvements in the flight code are presented. In addition, the DLC uses a commercial processor as a surrogate for NASA’s High-Performance Spaceflight Computing (HPSC) processor. This work provides insight on how an HPSC system may perform delivering PL&HA capabilities on a future mission. The measurements also can be used to infer architectural requirements for PL&HA capabilities, informing the HPSC project and other flight computer development efforts. Examples of the measurements collected include processor utilization, I/O bandwidth, cache and branch misses, and application profiles.

Precision Landing↗

Innovative Joining Method for Hybrid Composites with Tailored Performance

In various industries, such as renewables and aerospace, the demand for carbon fiber is rapidly growing to meet the requirements of high-performance applications with optimized designs. Traditionally, these optimized designs have been limited to single-material composites due to process constraints. However, this study aims to explore the feasibility of transitioning between two different fiber types for pultrusion and filament winding processes, thereby enhancing design flexibility, and optimizing performance. This research introduces a novel method of hybrid reinforcement through splicing techniques. By employing a splicing method, fibers are merged prior to infusion, resulting in a transition of higher strength compared to traditional joints. Moreover, this technique enables the creation of composites with variable compositions, allowing for the selective placement of properties in a process previously characterized by static properties. The objective of this paper is to evaluate the mechanical properties of dry fiber splicing and its impact on the resulting composite material. Through comprehensive analysis, we aim to provide insights into the feasibility and advantages of employing this innovative splicing technique in composite manufacturing processes.

Guzorek, Steven↗

Camelina circRNA landscape: Implications for gene regulation and fatty acid metabolism

Abstract Circular RNAs (circRNAs) are closed‐loop RNAs forming a covalent bond between their 3′ and 5′ ends, the back splice junction (BSJ), rendering them resistant to exonucleases and thus more stable compared to linear RNAs. Identification of circRNAs and distinction from their cognate linear RNA is only possible by sequencing the BSJ that is unique to the circRNA. CircRNAs are involved in the regulation of their cognate RNAs by increasing transcription rates, RNA stability, and alternative splicing. We have identified circRNAs from C. sativa that are associated with the regulation of germination, light response, and lipid metabolism. We sequenced light‐grown and etiolated seedlings after 5 or 7 days post‐germination and identified a total of 3447 circRNAs from 2763 genes. Most circRNAs originate from a single homeolog of the three subgenomes from allohexaploid camelina and correlate with higher ratios of alternative splicing of their cognate genes. A network analysis shows the interactions of select miRNA:circRNA:mRNAs for regulation of transcript stabilities where circRNA can act as a competing endogenous RNA. Several key lipid metabolism genes can generate circRNA, and we confirmed the presence of KASII circRNA as a true circRNA. CircRNA in camelina can be a novel target for breeding and engineering efforts.

Utley, Delecia [Department of Plant and Microbial ↗

Precise mapping of new group I introns in tRNA genes

Bacterial tRNA have been found interrupted at various positions in the anticodon loop by group I introns, in four types. The primary bioinformatic tool for group I intron discovery is a covariance model that can identify conserved features in the catalytic core and can sometimes identify the typical uridine residue at the -1 position, preceding the 5-prime splice site, but cannot identify the typical guanidine residue at the omega position, preceding the 3-prime splice site, to achieve precise mapping. One approach to complete the automation of group I intron mapping is to focus instead on the exons, which is enabled by the regularity of tRNAs. We develop a software module, within a larger package (tFind) aimed at mapping bacterial tRNA and tmRNA genes precisely, that expands this list of four known classes of intron-interrupted tRNAs to 21 cases. A new covariance model for these introns is presented. The wobble base pair formed by the -1 uridine is considered a determinant of the 5-prime splice site, yet one reasonably large new type bears a cytidine nucleotide at that position.

Williams, Kelly P.↗

SARS-CoV-2 Diverges from Other Betacoronaviruses in Only Partially Activating the IRE1α/XBP1 Endoplasmic Reticulum Stress Pathway in Human Lung-Derived Cells

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has killed over 6 million individuals worldwide and continues to spread in countries where vaccines are not yet widely available or its citizens are hesitant to become vaccinated. Therefore, it is critical to unravel the molecular mechanisms that allow SARS-CoV-2 and other coronaviruses to infect and overtake the host machinery of human cells. Coronavirus replication triggers endoplasmic reticulum (ER) stress and activation of the unfolded protein response (UPR), a key host cell pathway widely believed to be essential for viral replication. We examined the master UPR sensor IRE1α kinase/RNase and its downstream transcription factor effector XBP1s, which is processed through an IRE1α-mediated mRNA splicing event, in human lung-derived cells infected with betacoronaviruses. We found that human respiratory coronavirus OC43 (HCoV-OC43), Middle East respiratory syndrome coronavirus (MERS-CoV), and murine coronavirus (MHV) all induce ER stress and strongly trigger the kinase and RNase activities of IRE1α as well as XBP1 splicing. In contrast, SARS-CoV-2 only partially activates IRE1α through autophosphorylation, but its RNase activity fails to splice XBP1. Moreover, while IRE1α was dispensable for replication in human cells for all coronaviruses tested, it was required for maximal expression of genes associated with several key cellular functions, including the interferon signaling pathway, during SARS-CoV-2 infection. Our data suggest that SARS-CoV-2 actively inhibits the RNase of autophosphorylated IRE1α, perhaps as a strategy to eliminate detection by the host immune system.

60 APPLIED LIFE SCIENCES↗

Verification of the SRB motor case pin retainer band tips

The pin retainer band is a 1-1/4 in. wide by about 1/32 in. thick steel band that is stretched over the pins circumferentially at the joints of the solid rocket booster (SRB) motor case segments. Its purpose is to hold the retaining pins in place during flight. Since the band is thin and is mounted stretched, or under tension, over the pins which are greased in place, it is necessary to protect it thermally. Three typical areas of the band location on the SRB motor case joints were chosen for verifying the cork thermal prection system (TPS) performance on the band: the joint in front of the attach ring, at the aft skirt joint in front of the kick ring and in an unperturbed area at the forward segments of the SRB motor cases. The kick ring area pin retainer band cork TPS passed the test requirements in the region where there are no phenolic splice caps. In the splice cap areas where the band TPS is closed out around the splice with K5NA, additional amounts of K5NA material is required to adequately protect the steel band. TPS requirements were easily met in the one location where the ends of the steel band are joined together after installation and tensioning. The attack ring area pin retainer band TPS did not quite pass the heat load requirements with the .375 in. thick cork TPS as used for the kick ring area band. It is recommended that the cork thickness here be increased to .50 in. The unperturbed area band TPS presented no problems.

Karu, Z. S.↗

An instrumented fastener for shear force measurements in joints

A preliminary investigation has been conducted on instrumented fasteners for use as sensors to measure the shear loads transmitted by individual fasteners installed in double-splice joints. Calibration and load verification tests were conducted for instrumented fasteners installed at three fastener torque levels. Results from calibration tests show that the shear strains obtained from the instrumented fasteners vary linearly with the applied load and that the instrumented fasteners can be effectively used to measure shear loads transmitted by individual fasteners installed in double-splice joints. Tests were also conducted with three instumented fasteners installed in a typical double-splice joint. The test results showed that the load distribution between individual fasteners is dependent on the location of the fastener in the joint and the fastener torque level.

Sawyer, J. W.↗

Thermal Fault Tolerance Analysis of Carbon Fiber Rope Barrier Systems for Use in the Reusable Solid Rocket Motor ( RSRM) Nozzle Joints

Carbon Fiber Rope (CFR) thermal barrier systems are being considered for use in several RSRM (Reusable Solid Rocket Motor) nozzle joints as a replacement for the current assembly gap close-out process/design. This study provides for development and test verification of analysis methods used for flow-thermal modeling of a CFR thermal barrier subject to fault conditions such as rope combustion gas blow-by and CFR splice failure. Global model development is based on a 1-D (one dimensional) transient volume filling approach where the flow conditions are calculated as a function of internal 'pipe' and porous media 'Darcy' flow correlations. Combustion gas flow rates are calculated for the CFR on a per-linear inch basis and solved simultaneously with a detailed thermal-gas dynamic model of a local region of gas blow by (or splice fault). Effects of gas compressibility, friction and heat transfer are accounted for the model. Computational Fluid Dynamic (CFD) solutions of the fault regions are used to characterize the local flow field, quantify the amount of free jet spreading and assist in the determination of impingement film coefficients on the nozzle housings. Gas to wall heat transfer is simulated by a large thermal finite element grid of the local structure. The employed numerical technique loosely couples the FE (Finite Element) solution with the gas dynamics solution of the faulted region. All free constants that appear in the governing equations are calibrated by hot fire sub-scale test. The calibrated model is used to make flight predictions using motor aft end environments and timelines. Model results indicate that CFR barrier systems provide a near 'vented joint' style of pressurization. Hypothetical fault conditions considered in this study (blow by, splice defect) are relatively benign in terms of overall heating to nozzle metal housing structures.

Clayton, J. Louie↗

Defining Navigation Requirements for Future Precision Lander Missions

Human mission concepts for both the Moon and Mars require landing precision capabilities beyond the current state of art. The Safe and Precise Landing Integrated Capabilities Evolution (SPLICE) project leverages previous work at NASA to develop multimission precision landing and hazard avoidance (PL&HA) technologies to meet the advanced landing requirements. SPLICE aims to develop guidance, navigation and control (GN&C) capabilities for both Moon and Mars lander missions. The approach the project is using to identify a standard suite begins with developing a PL&HA requirements matrix to inform technology investments. The approach to develop a multi-mission navigation requirements matrix includes identifying representative concepts of operations for both robotic and human missions. It also identifies navigation sensor options with estimated performance parameters. This paper summarizes the concepts of operations and sensors considered to develop the SPLICE PL&HA requirements matrix. A description of the analysis used to determine candidate optimal sensor suites and initial results are provided.

Dwyer Cianciolo, Alicia↗

mTOR regulates aerobic glycolysis through NEAT1 and nuclear paraspeckle-mediated mechanism in hepatocellular carcinoma

Hepatocellular Carcinoma (HCC) is a major form of liver cancer and a leading cause of cancer-related death worldwide. New insights into HCC pathobiology and mechanism of drug actions are urgently needed to improve patient outcomes. HCC undergoes metabolic reprogramming of glucose metabolism from respiration to aerobic glycolysis, a phenomenon known as the ‘Warburg Effect’ that supports rapid cancer cell growth, survival, and invasion. mTOR is known to promote Warburg Effect, but the underlying mechanism(s) remains poorly defined. The aim of this study is to understand the mechanism(s) and significance of mTOR regulation of aerobic glycolysis in HCC. We profiled mTORC1-dependent long non-coding RNAs (lncRNAs) by RNA-seq of HCC cells treated with rapamycin. Chromatin immunoprecipitation (ChIP) and luciferase reporter assays were used to explore the transcriptional regulation of NEAT1 by mTORC1. [U- 13 C]-glucose labeling and metabolomic analysis, extracellular acidification Rate (ECAR) by Seahorse XF Analyzer, and glucose uptake assay were used to investigate the role of mTOR-NEAT1-NONO signaling in the regulation of aerobic glycolysis. RNA immunoprecipitation (RIP) and NONO-binding motif scanning were performed to identify the regulatory mechanism of pre-mRNA splicing by mTOR-NEAT1. Myristoylated AKT1 (mAKT1)/NRASV 12 -driven HCC model developed by hydrodynamic transfection (HDT) was employed to explore the significance of mTOR-NEAT1 signaling in HCC tumorigenesis and mTOR-targeted therapy. mTOR regulates lncRNA transcriptome in HCC and that NEAT1 is a major mTOR transcriptional target. Interestingly, although both NEAT1_1 and NEAT1_2 are down-regulated in HCC, only NEAT1_2 is significantly correlated with poor overall survival of HCC patients. NEAT1_2 is the organizer of nuclear paraspeckles that sequester the RNA-binding proteins NONO and SFPQ. We show that upon oncogenic activation, mTORC1 suppresses NEAT1_2 expression and paraspeckle biogenesis, liberating NONO/SFPQ, which in turn, binds to U5 within the spliceosome, stimulating mRNA splicing and expression of key glycolytic enzymes. This series of actions lead to enhanced glucose transport, aerobic glycolytic flux, lactate production, and HCC growth both in vitro and in vivo. Furthermore, the paraspeckle-mediated mechanism is important for the anticancer action of US FDA-approved drugs rapamycin/temsirolimus. These findings reveal a molecular mechanism by which mTOR promotes the ‘Warburg Effect’, which is important for the metabolism and development of HCC, and anticancer response of mTOR-targeted therapy.

59 BASIC BIOLOGICAL SCIENCES↗

Precision Landing Performance of a Human-Scale Lunar Lander Using a Generalized Simulation Framework

NASA has established goals of returning humans to Moon with an initial landing by 2024 and a subsequent sustained presence by 2028, which will require technological advances in spacecraft navigation to enable precision landing. The ability to assess the navigation performance of these new and existing technologies is critical to identifying areas of risk reduction and investment. To that end, the Safe and Precise Landing Integrated Capabilities Evolution (SPLICE) project has demonstrated that a detailed six degree-of-freedom integrated performance simulation framework can provide information on and assessment of expected navigation performance. This framework incorporates engineering models of the on-board spacecraft guidance, navigation, and control systems at varying levels of fidelity. Recent advances in the development of this integrated performance simulation permit running these systems “in-the-loop,” rather than assuming perfect knowledge of the spacecraft states. This development, coupled with fast simulation time and modularization of the various system models, enables a wide variety of system trades to be assessed at once. This paper presents a summary of the advances in the SPLICE simulation framework, updates to the spacecraft navigation models, and an application of the framework to characterize the precision landing performance of a human-scale lunar lander. A series of trade studies examining effects of ground state update qualities shows that given all other assumptions, sufficiently accurate Deep Space Network (DSN) measurements can enable safe and precise human-scale Lunar landings.

Spacecraft navigation↗