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At least 127 records · Page 7

Performance Analysis of A Dual Quaternion Guidance Algorithm Applicable During Lunar Approach With A Hazard Avoidance Maneuver

There is currently a need for advanced guidance and targeting algorithms that can provide real-time trajectories in the presence of state and vehicle constraints during powered descent. The Safe & Precise Landing Integrated Capabilities Evolution (SPLICE) program aims to mature Hazard Detection (HD) technology along with advanced navigation and guidance algorithms. This paper will report the overall performance of the SPLICE Dual Quaternion Guidance (DQG) algorithm which is a 6-Degree-of-Freedom (6-DOF) convex optimization-based guidance algorithm that can enforce multiple vehicle and state constraints required for Hazard Detection Lidar (HDL) scans during the approach phase. DQG was tested in a closed-loop lunar descent simulation with realistic HDL sensing constraints in a Monte Carlo assessment of 1000 dispersed runs.

Guidance↗

Long- and short-read sequencing methods discover distinct circular RNA pools in Lotus japonicus

Circular RNAs (circRNAs) are covalently closed single-stranded RNAs, generated through a back-splicing process that links a downstream 5′ site to an upstream 3′ end. The only distinction in the sequence between circRNA and their linear cognate RNA is the back splice junction. Their low abundance and sequence similarity with their linear origin RNA have made the discovery and identification of circRNA challenging. We have identified almost 6000 novel circRNAs from Lotus japonicus leaf tissue using different enrichment, amplification, and sequencing methods as well as alternative bioinformatics pipelines. The different methodologies identified different pools of circRNA with little overlap. We validated circRNA identified by the different methods using reverse transcription polymerase chain reaction and characterized sequence variations using nanopore sequencing. We compared validated circRNA identified in L. japonicus to other plant species and showed conservation of high-confidence circRNA-expressing genes. This is the first identification of L. japonicus circRNA and provides a resource for further characterization of their function in gene regulation. CircRNAs identified in this study originated from genes involved in all biological functions of eukaryotic cells. The comparison of methodologies and technologies to sequence, identify, analyze, and validate circRNA from plant tissues will enable further research to characterize the function and biogenesis of circRNA in L. japonicus.

59 BASIC BIOLOGICAL SCIENCES↗

Single-molecule epitranscriptomic analysis of full-length HIV-1 RNAs reveals functional roles of site-specific m6As

Abstract Although the significance of chemical modifications on RNA is acknowledged, the evolutionary benefits and specific roles in human immunodeficiency virus (HIV-1) replication remain elusive. Most studies have provided only population-averaged values of modifications for fragmented RNAs at low resolution and have relied on indirect analyses of phenotypic effects by perturbing host effectors. Here we analysed chemical modifications on HIV-1 RNAs at the full-length, single RNA level and nucleotide resolution using direct RNA sequencing methods. Our data reveal an unexpectedly simple HIV-1 modification landscape, highlighting three predominant N 6 -methyladenosine (m 6 A) modifications near the 3′ end. More densely installed in spliced viral messenger RNAs than in genomic RNAs, these m 6 As play a crucial role in maintaining normal levels of HIV-1 RNA splicing and translation. HIV-1 generates diverse RNA subspecies with distinct m 6 A ensembles, and maintaining multiple of these m 6 As on its RNAs provides additional stability and resilience to HIV-1 replication, suggesting an unexplored viral RNA-level evolutionary strategy.

60 APPLIED LIFE SCIENCES↗

An alternative domain-swapped structure of the Pyrococcus horikoshii PolII mini-intein

Protein splicing is a post-translational process by which an intein catalyzes its own excision from flanking polypeptides, or exteins, concomitant with extein ligation. Many inteins have nested homing endonuclease domains that facilitate their propagation into intein-less alleles, whereas other inteins lack the homing endonuclease (HEN) and are called mini-inteins. The mini-intein that interrupts the DNA PolII of Pyrococcus horikoshii has a linker region in place of the HEN domain that is shorter than the linker in a closely related intein from Pyrococcus abyssi. The P. horikoshii PolII intein requires a higher temperature for catalytic activity and is more stable to digestion by the thermostable protease thermolysin, suggesting that it is more rigid than the P. abyssi intein. We solved a crystal structure of the intein precursor that revealed a domain-swapped dimer. Inteins found as domain swapped dimers have been shown to promote intein-mediated protein alternative splicing, but the solved P. horikoshii PolII intein structure has an active site unlikely to be catalytically competent.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Post-Transcriptional Mechanisms Respond Rapidly to Ecologically Relevant Thermal Fluctuations During Temperature-Dependent Sex Determination

An organism’s ability to integrate transient environmental cues experienced during development into molecular and physiological responses forms the basis for adaptive shifts in phenotypic trajectories. During temperature-dependent sex determination (TSD), thermal cues during discrete periods in development coordinate molecular changes that ultimately dictate sexual fate and contribute to patterns of inter- and intra-sexual variation. How these mechanisms interface with dynamic thermal environments in nature remain largely unknown. By deploying thermal loggers in wild nests of the American alligator (Alligator mississippiensis) over two consecutive breeding seasons, we observed that 80% of nests exhibit both male- and female-promoting thermal cues during the thermosensitive period, and of these nests, all exhibited both male- and female-promoting temperatures within the span of a single day. These observations raise a critical question—how are opposing environmental cues integrated into sexually dimorphic transcriptional programs across short temporal scales? To address this question, alligator embryos were exposed to fluctuating temperatures based on nest thermal profiles and sampled over the course of a daily thermal fluctuation. We examined the expression dynamics of upstream genes in the temperature-sensing pathway and find that post-transcriptional alternative splicing and transcript abundance of epigenetic modifier genes JARID2 and KDM6B respond rapidly to thermal fluctuations while transcriptional changes of downstream effector genes, SOX9 and DMRT1, occur on a delayed timescale. Our findings reveal how the basic mechanisms of TSD operate in an ecologically relevant context. We present a hypothetical hierarchical model based on our findings as well as previous studies, in which temperature-sensitive alternative splicing incrementally influences the epigenetic landscape to affect the transcriptional activity of key sex-determining genes.

59 BASIC BIOLOGICAL SCIENCES↗

Light Propagation Considerations For Internally Clad Sapphire Optical Fiber Using the 6Li(n,)3H Reaction

Optical frequency domain reflectometry measurements in internally clad single crystal sapphire fiber have received attention in recent years due to their high temperature distributed sensing potential. As work with these fibers has proceeded, there have been some inconsistencies in the results. Deeper investigation and testing has identified two critical considerations for the proper functioning of these fibers. First, users must address the propagation of multimode light along the outer surface of the fiber. Here, by observing the far-field image of an internally clad sapphire fiber when adding index matching fluid to the outer fiber surface, we demonstrate the effects of removing the higher order modes from these fibers. The addition of index matching fluid resulted in nearly single mode performance where multimode performance was previously observed. Second, users must address the effect that coupling the fiber to the interrogator via silica based fiber has on the internally clad sapphire fibers performance. Direct fusion splicing of silica to sapphire, as has been used in the recent work with these fibers, has a mode filtering effect which can be beneficial towards the modal behavior of the fibers. However, in this paper we demonstrate that the splicing can cause a sensing failure due to little or no low order mode light, that is useful for sensing, returning to the detector. The positive results from recent years have demonstrated that optical frequency domain reflectometry sensing performance will be successful in clad sapphire fiber; but only when the considerations described herein are addressed properly.

47 OTHER INSTRUMENTATION↗

Expanding the genetic toolkit: adenine and cytosine base editors for gene disruption in Aspergillus niger

Despite revolutionizing fungal genetic engineering, conventional CRISPR/Cas9-mediated knockouts rely on DNA double-strand breaks (DSBs), which can cause unwanted insertions and deletions, chromosomal abnormalities, and cytotoxicity. Base editors such as adenine base editors (ABEs), which convert A‧T to G‧C, and cytosine base editors (CBEs), which convert C‧G to T‧A, offer a safer alternative by enabling predictable, target-specific single-nucleotide changes without introducing DSBs. To overcome the limitations of traditional genome editing in filamentous fungi, we developed efficient base-editing systems in Aspergillus niger . For the first time, we constructed an ABE in A. niger , achieving up to 80% editing efficiency and inducing predictable A-to-G mutations at the intended intron sites, disrupting gene function through mRNA mis-splicing. We also developed a highly efficient CBE system, capable of introducing premature stop codons with 50–100% efficiency. To broaden the editing scope, we implemented a Cas9-NG variant recognizing a relaxed PAM sequence requiring only a single guanine (G), enabling editing at start codons and splice sites. Leveraging this expanded scope, we established gene disruption approaches by targeting start codons via ABE-mediated A-to-G conversions (ATG-to-GTG and ATG-to-ACG) and CBE-mediated C-to-T conversion (ATG-to-ATA). Additionally, our base-editing systems enable multiplex gRNA delivery and marker-free editing of multiple genes. Collectively, the scope-expanding strategies increase the number of genes targetable for disruption by base-editing in A. niger by 26.3% and enable near-complete coverage of 96% of the coding genes. Overall, this work demonstrates the potential of ABE and CBE systems as versatile, efficient, and safer alternatives to DSBs-based gene disruption in filamentous fungi.

Aspergillus↗

Data for A Role for Differential Rubisco Activase Isoform Expression in C4 Bioenergy Grasses at High Temperature

Rubisco activase (Rca) facilitates the release of sugar-phosphate inhibitors at Rubisco catalytic sites during CO2 fixation. Most plant species express two Rca isoforms, the larger Rca-α and the shorter Rca-β, either by alternative splicing from a single gene or expression from separate genes. The mechanism of Rubisco activation by Rca isoforms has been intensively studied in C3 plants. However, the functional role of Rca in C4 plants where Rubisco and Rca are located in a much higher [CO2] compartment is less clear. In this study, we selected four C4 bioenergy grasses and the model C4 grass setaria ( Setaria viridis ) to investigate the role of Rca in C4 photosynthesis. All five C4 grass species contained two Rca genes, one encoding Rca-α and the other Rca-β, which were positioned closely together in the genomes. A variety of abiotic stress-related motifs were identified in the Rca-α promoter of each grass, and while the Rca-β gene was constantly highly expressed at ambient temperature, Rca-α isoforms were expressed only at high temperature but never surpassed 30% of Rca-β content. The pattern of Rca-α induction on transition to high temperature and reduction on return to ambient temperature was the same in all five C4 grasses. In sorghum ( Sorghum bicolor ), sugarcane ( Saccharum officinarum ), and setaria, the induction rate of Rca-α was similar to the recovery rate of photosynthesis and Rubisco activation at high temperature. This association between Rca-α isoform expression and maintenance of Rubisco activation at high temperature suggests that Rca-α has a functional thermo-protective role in carbon fixation in C4 grasses by sustaining Rubisco activation at high temperature.

Genomics↗

Distributed Acoustic Sensing (DAS) on Opportunistic Networks_ A Feasibility Study Utilizing Fiber Optic Infrastructure at NEON Sites for DAS

This study demonstrates the feasibility of using Distributed Acoustic Sensing (DAS) on the opportunistic optical fiber infrastructure presented by NEON (National Ecological Observatory Network) sites. The test took place at the NEON Terrestrial Site designated D10 CPER. Unlit portions of the local fiber network were successfully spliced together and optimized for length and a Silixa iDAS sensor was installed. The resulting array (composed of 12 spliced fiber runs) totaled 875m in length. Over a period of 2 weeks, we gathered a high spatial and temporal resolution data set to characterize discernable signals and understand background noise. The data set was reviewed to extract wave signatures of identifiable origin, both anthropogenic and otherwise. Continent wide distribution of these sites makes them an interesting candidate for DAS and identifying measurable signal sources informs future DAS application on opportunistic fiber networks. These observations validate the utility of both the NEON and similarly small fiber networks (<1km) for geoscience, environmental, and security DAS applications and indicate possibilities for further inquiry in methods for DAS data collection and analysis.

42 ENGINEERING↗

Study to investigate design, fabrication and test of low cost concepts for large hybrid composite helicopter fuselage, phase 1

The development of a frame/stringer/skin fabrication technique for composite airframe construction was studied as a low cost approach to the manufacture of large helicopter airframe components. A center cabin aluminum airframe section of the Sikorsky CH-53D helicopter was selected for evaluation as a composite structure. The design, as developed, is composed of a woven KEVLAR-49/epoxy skin and graphite/epoxy frames and stringers. To support the selection of this specific design concept a materials study was conducted to develop and select a cure compatible graphite and KEVLAR-49/epoxy resin system, and a foam system capable of maintaining shape and integrity under the processing conditions established. The materials selected were, Narmco 5209/Thornel T-300 graphite, Narmco 5209/KEVLAR-49 woven fabric, and Stathane 8747 polyurethane foam. Eight specimens were fabricated, representative of the frame, stringer, and splice joint attachments. Evaluation of the results of analysis and test indicate that design predictions are good to excellent except for some conservatism of the complex frame splice.

Adams, K. M.↗

Reliability aspects of a composite bolted scarf joint

The design, fabrication, static test, and fatigue test of both tension and compression graphite-epoxy candidates for a wing splice representative of a next-generation transport aircraft was the objective of the reported research program. A single-scarf bolted joint was selected as the design concept. Test specimens were designed and fabricated to represent an upper-surface and a lower-surface panel containing the splice. The load spectrum was a flight-by-flight random-load history including ground-air-ground loads. The results of the fatigue testing indicate that, for this type of joint, the inherent fatigue resistance of the laminate is reflected in the joint behavior and, consequently, the rate of damage accumulation is very slow under realistic fatigue loadings.

Reed, D. L.↗

Strain gage system evaluation program

A program was conducted to determine the reliability of various strain gage systems when applied to rotating compressor blades in an aircraft gas turbine engine. A survey of current technology strain gage systems was conducted to provide a basis for selecting candidate systems for evaluation. Testing and evaluation was conducted in an F 100 engine. Sixty strain gage systems of seven different designs were installed on the first and third stages of an F 100 engine fan. Nineteen strain gage failures occurred during 62 hours of engine operation, for a survival rate of 68 percent. Of the failures, 16 occurred at blade-to-disk leadwire jumps (84 percent), two at a leadwire splice (11 percent), and one at a gage splice (5 percent). Effects of erosion, temperature, G-loading, and stress levels are discussed. Results of a post-test analysis of the individual components of each strain gage system are presented.

Dolleris, G. W.↗

Optimized bolted joint

A method is disclosed for joining segments of the skin of an aircraft. The ends of the skin are positioned in close proximity or abutt each other. The skin is of constant thickness throughout the joint and is sandwiched between splice plates, which taper in thickness from the last to the first bolt rows in order to reduce the stiffness of the splice plate and thereby reduce the load transfer at the location where bypass loads are the highest.

Hart-Smith, L. J.↗

Critical joints in large composite primary aircraft structures. Volume 2: Technology demonstration test report

A program was conducted to develop the technology for critical structural joints in composite wing structure that meets all the design requirements of a 1990 commercial transport aircraft. The results of four large composite multirow bolted joint tests are presented. The tests were conducted to demonstrate the technology for critical joints in highly loaded composite structure and to verify the analytical methods that were developed throughout the program. The test consisted of a wing skin-stringer transition specimen representing a stringer runout and skin splice on the wing lower surface at the side of the fuselage attachment. All tests were static tension tests. The composite material was Toray T-300 fiber with Ciba-Geigy 914 resin in 10 mil tape form. The splice members were metallic, using combinations of aluminum and titanium. Discussions are given of the test article, instrumentation, test setup, test procedures, and test results for each of the four specimens. Some of the analytical predictions are also included.

Bunin, Bruce L.↗

Sapphire-fiber-based intrinsic Fabry-Perot interferometer

A sapphire optical fiber intrinsic Fabry-Perot interferometric sensor is demonstrated. A length of multimode sapphire fiber that functions as a Fabry-Perot cavity is spliced to a silica single-mode fiber. The interferometric signals of this sensor are produced by the interference between the reflection from the silica-sapphire fiber splice and the reflection from the free end face of the sapphire fiber. This sensor has been demonstrated for temperature measurement. A resolution of 0.2 C has been obtained over a measurement range of 310 C to 976 C.

Wang, Anbo↗

Advances in sapphire optical fiber sensors

We describe the development and testing of two sapphire fiber sensor designs intended for use in high temperature environments. The first is a birefringence-balanced polarimetric sapphire fiber sensor. In this sensor, two single crystal sapphire rods, acting as the birefringence sensing element, are connected to each other in such a way that the slow axis of the first rod is aligned along with the fast axis of the second rod, and the fast axis of the first rod is along the slow axis of the second rod. This sensor has been demonstrated for measurement of temperature up to 1500 C. The second is a sapphire-fiber-based intrinsic interferometric sensor. In this sensor, a length of uncoated, unclad, structural-graded multimode sapphire fiber is fusion spliced to a singlemode silica fiber to form a Fabry-Perot cavity. The reflections from the silica-to-sapphire fiber splice and the free endface of the sapphire fiber give rise to the interfering fringe output. This sensor has been demonstrated for the measurement of temperature above 1510 C, and a resolution of 0.1 C has been obtained.

Wang, Anbo↗

Demonstrating damage tolerance of composite airframes

Commercial transport aircraft operating in the United States are certified by the Federal Aviation Authority to be damage tolerant. On 28 April 1988, Aloha Airlines Flight 243, a Boeing 727-200 airplane, suffered an explosive decompression of the fuselage but landed safely. This event provides very strong justification for the damage tolerant design criteria. The likely cause of the explosive decompression was the linkup of numerous small fatigue cracks that initiated at adjacent fastener holes in the lap splice joint at the side of the body. Actually, the design should have limited the damage size to less than two frame spacings (about 40 inches), but this type of 'multi-site damage' was not originally taken into account. This cracking pattern developed only in the high-time airplanes (many flights). After discovery in the fleet, a stringent inspection program using eddy current techniques was inaugurated to discover these cracks before they linked up. Because of concerns about safety and the maintenance burden, the lap-splice joints of these high-time airplanes are being modified to remove cracks and prevent new cracking; newer designs account for 'multi-site damage'.

Poe, Clarence C., Jr.↗

An instrumented fastener for shear force measurements in joints

Results are presented of a preliminary investigation of 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. Calibration test results 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. The load distribution between individual fasteners is found to be dependent on the location of the fastener in the joint and the fastener torque level.

Sawyer, James W.↗