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All sources NASA NTRS DOE OSTI

DOE OSTI2026

Data for A Fluorescence-Based Transient Expression Assay for the Analysis of Upstream Open Reading Frames in Plants

Scripts for the manuscript "A fluorescence-based transient expression assay for the analysis of upstream open reading frames in plant" by Haas et al. Upstream open reading frames (uORFs) are regulatory elements present in the 5′ leaders of mRNA that can significantly impact downstream gene expression in eukaryotes. In crop engineering, editing of uORFs can provide an avenue to upregulate expression of native genes without the need to add persistent transgenic copies. Even with genome- wide methods to identify translated uORFs such as ribosome profiling, their functional characterization depends on validation through reporter gene assays and mutagenesis studies. Current screening methods for plants use luciferases or protoplasts to measure differential gene expression between wild- type and mutated transcript leaders, which requires tissue processing and/or substrate addition. Here, we present a time- and cost- efficient alternative to investigate transcript leaders by co- expression of two fluorescent proteins in Nicotiana benthamiana leaf tissue and test our assay on genes involved in photoprotection, editing of which could provide a pathway to increase CO2 assimilation during sun–shade transitions.

Gene Editing↗

DOE OSTI2026

Human RNome Project draft human RNome sequence of GM12878, B-cell line, obtained by mass-spectrometry sequencing, long-read sequencing and short-read sequencing.

Here we report the first draft of the human RNome sequence, a reference map of RNA chemical modifications in a human B-cell line. RNA carries a diverse repertoire of chemical modifications that regulate gene expression, cellular function, and responses to physiological and pathological cues. Yet, unlike the genome, no reference map of RNA modifications is available for any human cell. To generate this resource, the Human RNome Project Consortium analyzed a shared RNA preparation from the well-characterized GM12878 B-cell line using short-read sequencing, long-read direct RNA sequencing, and mass spectrometry, generating more than 7.1 billion sequencing reads spanning approximately 1.2 trillion nucleotides. The resulting maps of the human RNome reveal that RNA modifications are organized according to function, transcript architecture, and cellular identity. Modifications concentrate at functional centers of ribosomal and transfer RNAs, follow the canonical topology of N6-methyladenosine in coding transcripts, and form coordinated hotspots in immune regulatory genes. This first reference human RNome provides a foundation for understanding how RNA chemistry shapes cellular identity, human disease, and the development of RNA-based therapeutics.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2024

Evaluating two live-attenuated vaccines against Salmonella enterica serovar Reading in turkeys: reduced tissue colonization and cecal tonsil transcriptome responses

Vaccines that cross-protect across serovars ofSalmonella enterica(Salmonella) would be a beneficial intervention against emerging and persistentSalmonellaisolates of concern for the turkey industry. The 2017–2019 foodborne outbreak ofSalmonella entericaserovar Reading (S. Reading) revealed the need for effective control of this serovar in turkey production. This study evaluated two live-attenuatedSalmonellavaccines, an internally developed cross-protective vaccine and a commercially available vaccine, against an outbreak-associated strain ofS. Reading in turkeys. At 1 day and 3 weeks of age, male turkey poults were either mock-vaccinated with phosphate buffered saline (PBS) or given one of the vaccines by oral gavage (primary and booster) or aerosol spray (primary) then drinking water (booster). At 7 weeks of age, poults were challenged with 10 9 colony forming units (CFU) ofS. Reading; a mock-vaccinated group was mock-challenged with PBS. Colonization of the cecal contents and cecal tonsil was 1.5–3 log 10 CFU/g lower in vaccinated birds than mock-vaccinated birds at 7 and/or 14 days post-inoculation (DPI).Salmonelladissemination to the spleen was significantly reduced by both vaccines. Gene expression of intestinal transporters (such asSCNN1BandSLC10A2) and tight junction proteins was significantly decreased in the turkey cecal tonsil transcriptome at 2 DPI withS. Reading. Vaccination with either vaccine mitigated most cecal tonsil gene expression responses toS. Reading challenge. Therefore, both the internally developed vaccine and commercial vaccine were cross-protective against colonization and dissemination, and both were able to limit transcriptional changes from challenge in intestinal health-related genes in the cecal tonsil, thereby providing vaccination efficacy and impact data againstS. Reading in turkeys.

Veterinary Sciences↗

NASA NTRS1993

Read buffer optimizations to support compiler-assisted multiple instruction retry

Multiple instruction retry is a recovery mechanism for transient processor faults. We previously developed a compiler-assisted approach to multiple instruction ferry in which a read buffer of size 2N (where N represents the maximum instruction rollback distance) was used to resolve some data hazards while the compiler resolved the remaining hazards. The compiler-assisted scheme was shown to reduce the performance overhead and/or hardware complexity normally associated with hardware-only retry schemes. This paper examines the size and design of the read buffer. We establish a practical lower bound and average size requirement for the read buffer by modifying the scheme to save only the data required for rollback. The study measures the effect on the performance of a DECstation 3100 running ten application programs using six read buffer configurations with varying read buffer sizes. Two alternative configurations are shown to be the most efficient and differed depending on whether split-cycle-saves are assumed. Up to a 55 percent read buffer size reduction is achievable with an average reduction of 39 percent given the most efficient read buffer configuration and a variety of applications.

Alewine, N. J.↗

NASA NTRS1995

The Empirical Investigation of Perspective-Based Reading

We consider reading techniques a fundamental means of achieving high quality software. Due to lack of research in this area, we are experimenting with the application and comparison of various reading techniques. This paper deals with our experiences with Perspective Based Reading (PBR) a particular reading technique for requirement documents. The goal of PBR is to provide operation scenarios where members of a review team read a document from a particular perspective (eg., tester, developer, user). Our assumption is that the combination of different perspective provides better coverage of the document than the same number of readers using their usual technique. To test the efficacy of PBR, we conducted two runs of a controlled experiment in the environment of NASA GSFC Software Engineering Laboratory (SEL), using developers from the environment. The subjects read two types of documents, one generic in nature and the other from the NASA Domain, using two reading techniques, PBR and their usual technique. The results from these experiment as well as the experimental design, are presented and analyzed. When there is a statistically significant distinction, PBR performs better than the subjects' usual technique. However, PBR appears to be more effective on the generic documents than on the NASA documents.

Basili, Victor R.↗

NASA NTRS1992

Improved accuracy in ground-based facilities - Development of an automated film-reading system for ballistic ranges

Software for an automated film-reading system that uses personal computers and digitized shadowgraphs is described. The software identifies pixels associated with fiducial-line and model images, and least-squares procedures are used to calculate the positions and orientations of the images. Automated position and orientation readings for sphere and cone models are compared to those obtained using a manual film reader. When facility calibration errors are removed from these readings, the accuracy of the automated readings is better than the pixel resolution, and it is equal to, or better than, the manual readings. The effects of film-reading and facility-calibration errors on calculated aerodynamic coefficients is discussed.

Yates, Leslie A.↗

NASA NTRS1992

Development of an automated film-reading system for ballistic ranges

Yates, Leslie A.↗

NASA NTRS2015

Results from On-Board CSA-CP and CDM Sensor Readings During the Burning and Suppression of Solids II (BASS-II) Experiment in the Microgravity Science Glovebox (MSG)

For the first time on ISS, BASS-II utilized MSG working volume dilution with gaseous nitrogen (N2). We developed a perfectly stirred reactor model to determine the N2 flow time and flow rate to obtain the desired reduced oxygen concentration in the working volume for each test. We calibrated the model with CSA-CP oxygen readings offset using the Mass Constituents Analyzer reading of the ISS ambient atmosphere data for that day. This worked out extremely well for operations, and added a new vital variable, ambient oxygen level, to our test matrices. The main variables tested in BASS-II were ambient oxygen concentration, ventilation flow velocity, and fuel type, thickness, and geometry. BASS-II also utilized the on-board CSA-CP for oxygen and carbon monoxide readings, and the CDM for carbon dioxide readings before and after each test. Readings from these sensors allow us to evaluate the completeness of the combustion. The oxygen and carbon dioxide readings before and after each test were analyzed and compared very well to stoichiometric ratios for a one step gas-phase reaction. The CO versus CO2 followed a linear trend for some datasets, but not for all the different geometries of fuel and flow tested. Lastly, we calculated the heat release rates during each test from the oxygen consumption and burn times, using the constant 13.1 kJ of heat released per gram of oxygen consumed. The results showed that the majority of the tests had heat release rates well below 100 Watts.

combustion products↗

DOE OSTI2025

Calibration of multisite raters for prospective visual reads of amyloid PET scans

Abstract INTRODUCTION In multicenter Alzheimer's disease studies, amyloid positron emission tomography (PET) visual reads are typically performed centrally by a few experts. Incorporating a broader reader network enhances scalability and generalizability. METHODS Ten neuroimaging experts from eight Alzheimer's Disease Research Centers (ADRCs) visually read 180 amyloid PET scans (30 scans and 15 duplicate scans for each of four tracers, imaged across a wide variety of scanners), using preferred reading software without anatomical imaging or quantitation. Scans were classified as elevated or non‐elevated per tracer‐specific criteria. Inter‐ and intra‐rater agreement was assessed. RESULTS Inter‐rater agreement was substantial (Fleiss’κ = 0.78), with full consensus on 69% of scans. Inter‐rater reliability was substantial to perfect across tracers (Fleiss’κ = 0.70–0.87). Intra‐rater agreement was substantial to perfect (Cohen'sκ = 0.79‐1). Scans with intermediate (10–40 Centiloid) quantitation had lower reader agreement. DISCUSSION A multicenter expert network achieved substantial agreement classifying amyloid PET scans. These scans provide a standard for reader training and reliability assurance in future studies. Highlights Calibration methods ensure reliable amyloid positron emission tomography (PET) visual reads across multiple raters. Substantial agreement is possible across readers using their preferred tools. Agreement is also substantial regardless of the amyloid PET tracer used. Scans with intermediate (10–40 Centiloid) quantitation have lower reader agreement. The calibration set will become a training tool for amyloid PET visual read studies.

Neurosciences & Neurology↗

DOE OSTI2024

Decomposing a San Francisco estuary microbiome using long-read metagenomics reveals species- and strain-level dominance from picoeukaryotes to viruses

ABSTRACT Although long-read sequencing has enabled obtaining high-quality and complete genomes from metagenomes, many challenges still remain to completely decompose a metagenome into its constituent prokaryotic and viral genomes. This study focuses on decomposing an estuarine metagenome to obtain a more accurate estimate of microbial diversity. To achieve this, we developed a new bead-based DNA extraction method, a novel bin refinement method, and obtained 150 Gbp of Nanopore sequencing. We estimate that there are ~500 bacterial and archaeal species in our sample and obtained 68 high-quality bins (>90% complete, <5% contamination, ≤5 contigs, contig length of >100 kbp, and all ribosomal and tRNA genes). We also obtained many contigs of picoeukaryotes, environmental DNA of larger eukaryotes such as mammals, and complete mitochondrial and chloroplast genomes and detected ~40,000 viral populations. Our analysis indicates that there are only a few strains that comprise most of the species abundances. IMPORTANCE Ocean and estuarine microbiomes play critical roles in global element cycling and ecosystem function. Despite the importance of these microbial communities, many species still have not been cultured in the lab. Environmental sequencing is the primary way the function and population dynamics of these communities can be studied. Long-read sequencing provides an avenue to overcome limitations of short-read technologies to obtain complete microbial genomes but comes with its own technical challenges, such as needed sequencing depth and obtaining high-quality DNA. We present here new sampling and bioinformatics methods to attempt decomposing an estuarine microbiome into its constituent genomes. Our results suggest there are only a few strains that comprise most of the species abundances from viruses to picoeukaryotes, and to fully decompose a metagenome of this diversity requires 1 Tbp of long-read sequencing. We anticipate that as long-read sequencing technologies continue to improve, less sequencing will be needed.

Lui, Lauren M.↗

DOE OSTI2025

Long-read sequencing transcriptome quantification with lr-kallisto

RNA abundance quantification has become routine and affordable thanks to high-throughput “short-read” technologies that provide accurate molecule counts at the gene level. Similarly accurate and affordable quantification of definitive full-length, transcript isoforms has remained a stubborn challenge, despite its obvious biological significance across a wide range of problems. “Long-read” sequencing platforms now produce data-types that can, in principle, drive routine definitive isoform quantification. However some particulars of contemporary long-read datatypes, together with isoform complexity and genetic variation, present bioinformatic challenges. We show here, using ONT data, that fast and accurate quantification of long-read data is possible and that it is improved by exome capture. To perform quantifications we developed lr-kallisto, which adapts the kallisto bulk and single-cell RNA-seq quantification methods for long-read technologies.

Loving, Rebekah K. (ORCID:0000000187250376)↗

NASA NTRS1996

The Empirical Investigation of Perspective-Based Reading

We consider reading techniques a fundamental means of achieving high quality software. Due to the lack of research in this area, we are experimenting with the application and comparison of various reading techniques. This paper deals with our experiences with Perspective-Based Reading (PBR), a particular reading technique for requirements documents. The goal of PBR is to provide operational scenarios where members of a review team read a document from a particular perspective (e.g., tester, developer, user). Our assumption is that the combination of different perspectives provides better coverage of the document than the same number of readers using their usual technique.

Basili, Victor R.↗

NASA NTRS2003

Software for Automated Reading of STEP Files by I-DEAS(trademark)

A program called "readstep" enables the I-DEAS(tm) computer-aided-design (CAD) software to automatically read Standard for the Exchange of Product Model Data (STEP) files. (The STEP format is one of several used to transfer data between dissimilar CAD programs.) Prior to the development of "readstep," it was necessary to read STEP files into I-DEAS(tm) one at a time in a slow process that required repeated intervention by the user. In operation, "readstep" prompts the user for the location of the desired STEP files and the names of the I-DEAS(tm) project and model file, then generates an I-DEAS(tm) program file called "readstep.prg" and two Unix shell programs called "runner" and "controller." The program "runner" runs I-DEAS(tm) sessions that execute readstep.prg, while "controller" controls the execution of "runner" and edits readstep.prg if necessary. The user sets "runner" and "controller" into execution simultaneously, and then no further intervention by the user is required. When "runner" has finished, the user should see only parts from successfully read STEP files present in the model file. STEP files that could not be read successfully (e.g., because of format errors) should be regenerated before attempting to read them again.

Pinedo, John↗

NASA NTRS1984

Report-reading patterns of technical managers and nonmanagers

A survey to determine the review and reading processes used by technical managers and nonmanagers indicates that the summary, abstract, conclusion, title page, and introduction are the components used most frequently by both groups to decide whether to read a NASA technical report. In the review process, significantly more managers than nonmanagers use the summary and conclusion, whereas significantly more nonmanagers use the abstract and title page. The most common sequence of review consists of the title page, abstract, and summary, in that order, for both groups. In the reading process, the conclusion, results and discussion, and summary are the components read by the highest percentage of both groups.

Pinelli, T. E.↗

NASA NTRS1984

The Explorer's Guide to the Universe: A Reading List for Planetary and Space Science

During the last decade, both scientists and the public have been engulfed by a flood of discoveries and information from outer space. Distant worlds have become familiar landscapes. Instruments in space have shown us a different Sun by the "light" of ultraviolet radiation and X-rays. Beyond the solar system, we have detected a strange universe of unsuspected violence, unexplained objects, and unimaginable energies. We are completely remarking our picture of the universe around us, and scientists and the general public alike are curious and excited about what we see. The public has participated in this period of exploration and discovery to an extent never possible before. In real time, TV screens show moonwalks, the sands of Mars, the volcanoes of Io, and the rings of Saturn. But after the initial excitement, it is hard for the curious non-scientist to learn more details or even to stay in touch with what is going on. Each space mission or new discovery is quickly skimmed over by newspapers and TV and then preserved in technical journals that are neither accessible nor easily read by the average reader. This reading list is an attempt to bridge the gap between the people who make discoveries in space and the people who would like to read about them. The aim has been to provide to many different people--teachers, students, scientists, other professionals, and curious citizens of all kinds--a list of readings where they can find out what the universe is like and what we have learned about it. We have included sections on the objects that seem to be of general interest--the Moon, the planets, the Sun, comets, and the universe beyond. We have also included material on related subjects that people are interested in--the history of space exploration, space habitats, extraterrestrial life, and U F O ' s . The list is intended to be self-contained; it includes both general references to supply background and more specific sources for new discoveries. Although the list can be used by a wide range of people, it has been compiled with the non-scientist specifically in mind. A s a general rule, we have not included technical symposia or the special issues of

French, Bevan M.↗

NASA NTRS2004

Clarissa Spoken Dialogue System for Procedure Reading and Navigation

Speech is the most natural modality for humans use to communicate with other people, agents and complex systems. A spoken dialogue system must be robust to noise and able to mimic human conversational behavior, like correcting misunderstandings, answering simple questions about the task and understanding most well formed inquiries or commands. The system aims to understand the meaning of the human utterance, and if it does not, then it discards the utterance as being meant for someone else. The first operational system is Clarissa, a conversational procedure reader and navigator, which will be used in a System Development Test Objective (SDTO) on the International Space Station (ISS) during Expedition 10. In the present environment one astronaut reads the procedure on a Manual Procedure Viewer (MPV) or paper, and has to stop to read or turn pages, shifting focus from the task. Clarissa is designed to read and navigate ISS procedures entirely with speech, while the astronaut has his eyes and hands engaged in performing the task. The system also provides an MPV like graphical interface so the procedure can be read visually. A demo of the system will be given.

Hieronymus, James↗

NASA NTRS2009

Real-Time Detection of Dust Devils from Pressure Readings

A method for real-time detection of dust devils at a given location is based on identifying the abrupt, temporary decreases in atmospheric pressure that are characteristic of dust devils as they travel through that location. The method was conceived for use in a study of dust devils on the Martian surface, where bandwidth limitations encourage the transmission of only those blocks of data that are most likely to contain information about features of interest, such as dust devils. The method, which is a form of intelligent data compression, could readily be adapted to use for the same purpose in scientific investigation of dust devils on Earth. In this method, the readings of an atmospheric- pressure sensor are repeatedly digitized, recorded, and processed by an algorithm that looks for extreme deviations from a continually updated model of the current pressure environment. The question in formulating the algorithm is how to model current normal observations and what minimum magnitude deviation can be considered sufficiently anomalous as to indicate the presence of a dust devil. There is no single, simple answer to this question: any answer necessarily entails a compromise between false detections and misses. For the original Mars application, the answer was sought through analysis of sliding time windows of digitized pressure readings. Windows of 5-, 10-, and 15-minute durations were considered. The windows were advanced in increments of 30 seconds. Increments of other sizes can also be used, but computational cost increases as the increment decreases and analysis is performed more frequently. Pressure models were defined using a polynomial fit to the data within the windows. For example, the figure depicts pressure readings from a 10-minute window wherein the model was defined by a third-degree polynomial fit to the readings and dust devils were identified as negative deviations larger than both 3 standard deviations (from the mean) and 0.05 mbar in magnitude. An algorithm embodying the detection scheme of this example was found to yield a miss rate of just 8 percent and a false-detection rate of 57 percent when evaluated on historical pressure-sensor data collected by the Mars Pathfinder lander. Since dust devils occur infrequently over the course of a mission, prioritizing observations that contain successful detections could greatly conserve bandwidth allocated to a given mission. This technique can be used on future Mars landers and rovers, such as Mars Phoenix and the Mars Science Laboratory.

Wagstaff, Kiri↗

DOE OSTI2025

Long-read RNA sequencing atlas of human microglia isoforms elucidates disease-associated genetic regulation of splicing

Microglia, the innate immune cells of the central nervous system, have been genetically implicated in multiple neurodegenerative diseases. Mapping the genetics of gene expression in human microglia has identified several loci associated with disease-associated genetic variants in microglia-specific regulatory elements. However, identifying genetic effects on splicing is challenging because of the use of short sequencing reads. Here, we present the isoform-centric microglia genomic atlas (isoMiGA), which leverages long-read RNA sequencing to identify 35,879 novel microglia isoforms. We show that these isoforms are involved in stimulation response and brain region specificity. We then quantified the expression of both known and novel isoforms in a multi-ancestry meta-analysis of 555 human microglia short-read RNA sequencing samples from 391 donors, and found associations with genetic risk loci in Alzheimer’s and Parkinson’s disease. We nominate several loci that may act through complex changes in isoform and splice-site usage.

59 BASIC BIOLOGICAL SCIENCES↗