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At least 631 records · Page 35

Genomic Analysis of Caldithrix abyssi , the Thermophilic Anaerobic Bacterium of the Novel Bacterial Phylum Calditrichaeota

The genome of Caldithrix abyssi , the first cultivated representative of a phylum-level bacterial lineage, was sequenced within the framework of Genomic Encyclopedia of Bacteria and Archaea (GEBA) project. The genomic analysis revealed mechanisms allowing this anaerobic bacterium to ferment peptides or to implement nitrate reduction with acetate or molecular hydrogen as electron donors. The genome encoded five different [NiFe]- and [FeFe]-hydrogenases, one of which, group 1 [NiFe]-hydrogenase, is presumably involved in lithoheterotrophic growth, three other produce H 2 during fermentation, and one is apparently bidirectional. The ability to reduce nitrate is determined by a nitrate reductase of the Nap family, while nitrite reduction to ammonia is presumably catalyzed by an octaheme cytochrome c nitrite reductase εHao. The genome contained genes of respiratory polysulfide/thiosulfate reductase, however, elemental sulfur and thiosulfate were not used as the electron acceptors for anaerobic respiration with acetate or H 2 , probably due to the lack of the gene of the maturation protein. Nevertheless, elemental sulfur and thiosulfate stimulated growth on fermentable substrates (peptides), being reduced to sulfide, most probably through the action of the cytoplasmic sulfide dehydrogenase and/or NAD(P)-dependent [NiFe]-hydrogenase (sulfhydrogenase) encoded by the genome. Surprisingly, the genome of this anaerobic microorganism encoded all genes for cytochrome c oxidase, however, its maturation machinery seems to be non-operational due to genomic rearrangements of supplementary genes. Despite the fact that sugars were not among the substrates reported when C. abyssi was first described, our genomic analysis revealed multiple genes of glycoside hydrolases, and some of them were predicted to be secreted. This finding aided in bringing out four carbohydrates that supported the growth of C. abyssi : starch, cellobiose, glucomannan and xyloglucan. The genomic analysis demonstrated the ability of C. abyssi to synthesize nucleotides and most amino acids and vitamins. Finally, the genomic sequence allowed us to perform a phylogenomic analysis, based on 38 protein sequences, which confirmed the deep branching of this lineage and justified the proposal of a novel phylum Calditrichaeota .

59 BASIC BIOLOGICAL SCIENCES↗

Geology and Hydrothermal Alteration of the Raft River Geothermal System, Idaho

The Raft River geothermal system is located in southern Idaho, near the Utah-Idaho state boarder in the Raft River Valley. The field, which is owned and operated by U.S. Geothermal, has been selected as an EGS demonstration site by the U. S. Department of Energy. This paper summarizes ongoing geologic and petrologic investigations being conducted in support of this project. The reservoir is developed in fractured Proterozoic schist and quartzite, and Archean quartz monzonite cut by younger diabase intrusions. The basement complex was deformed during the mid Tertiary and covered by approximately 5000 ft of late Tertiary sedimentary and volcanic deposits. Listric normal faults of Cenozoic age disrupt the Tertiary deposits but do not offset the basement rocks. RRG-9, the target well, was drilled southwest of the main well field to a measured depth (MD) of 6089 ft. The well is deviated to the west and cased to a depth of 2316 ft MD. It penetrated the Proterozoic reservoir rocks at a depth of 5286 ft MD and encountered a maximum temperature of 139 degrees C. Bottomhole temperatures in other deep wells range from 133 to 149 degrees C. X-ray diffraction and thin section analyses are being conducted on samples from 5 deep wells, RRG- 1, 2, 3, 7 and 9, to determine the characteristics of the rock types and hydrothermal alteration within the geothermal system. Thin section analyses of samples from RRG-9 document the presence of strong alteration and brecciation at the contact between the Tertiary and basement complex. The Tertiary rocks consist of ash-flow tuffs, lava flows, tuffaceous siltstone, greywacke, and sandstone. No core is available from RRG-9 but core was obtained from RRG-3C. The sample is a brecciated and altered siltstone from the base of the Tertiary sequence and is similar to rocks at the base of the Tertiary deposits in RRG-9. Here, the results of thermal and quasi-static mechanical property measurements that were conducted on the core sample are presented.

Jones, Clay↗

Broadscale Ecological Patterns Are Robust to Use of Exact Sequence Variants versus Operational Taxonomic Units

Microbial ecologists have made exceptional improvements in our understanding of microbiomes in the last decade due to breakthroughs in sequencing technologies. These advances have wide-ranging implications for fields ranging from agriculture to human health. Due to limitations in databases, the majority of microbial ecology studies use a binning approach to approximate taxonomy based on DNA sequence similarity. There remains extensive debate on the best way to bin and approximate this taxonomy. Here we examine two popular approaches using a large field-based data set examining both bacteria and fungi and conclude that there are not major differences in the ecological outcomes. Thus, it appears that standard microbial community analyses are not overly sensitive to the particulars of binning approaches.

59 BASIC BIOLOGICAL SCIENCES↗

Congruence of Clusters Defined By Whole Genome Sequencing and MALDI-TOF for Bacteria Isolated From Cleanrooms

Introduction: Oligotrophic conditions can render cleanrooms inhospitable to microbes. Despite these constraints, fungi and bacteria are frequently isolated from surfaces in astromaterials cleanrooms at the Johnson Space Center. Bacillus species are of particular concern because endospores belonging to this genus are resilient and can affect astromaterials. Current monitoring programs rely on 16S rRNA sequencing and the VITEK2 Compact system. These methods have limited power to resolve Bacillus species. Matrix-assisted laser desorption - time of flight mass spectrometry (MALDI-TOF MS), provides a rapid, low cost, method of identifying bacterial isolates and has a higher resolution than 16S rRNA sequencing, particularly for Bacillus species; however, few studies have compared this method to the industry gold standard, whole genome sequencing (WGS). Methods: Based on 16S rRNA classification, we selected 14 isolates for analysis with MALDI-TOF and WGS. Mass spectra were generated with MALDI-TOF MS and processed with custom scripts to identify clusters of closely related isolates and calculate a matrix of pairwise cosine similarity scores. Hybrid Illumina and Nanopore sequencing were used to generate draft genomes. Pairwise similarity scores were calculated from these genomes based on the average amino acid identity (AAI) predicted from single copy core genes. Congruence of clustering between these methods, was assessed by calculating adjusted Rand and Wallace coefficients. Results: Clusters of species generated from MALDI-TOF MS showed good agreement of phylotypes generated with WGS. Pairs of strains that were > 94% similar to each other in terms of predicted amino acid sequences consistently showed cosine similarities of mass spectra > 0.65 and, of the 9 clusters identified with WGS, 8 were identical with MALDI-TOF. This corresponds to an adjusted Rand index of 0.95 and a 95% confidence interval of 0.80 – 1.00 for adjusted Wallace coefficients. The only discordance was for a pair of isolates that were classified as Paenibacillus species. This pair showed relatively high similarity (0.84) in terms of MALDI-TOF MS but only 85% similarity in terms of AAI. Conclusion: This study shows that MALDI-TOF and WGS exhibit a similar ability to delineate Bacillus species isolated from cleanrooms and taxonomic units described by these two methods are consistent with one another. Since MALDI-TOF MS is low in cost and high in throughput, this approach appears to be an ideal option for routine microbial monitoring and identifying Bacillus species.

Farnaz Mazhari↗

Comparison of motion and stereo methods in passive ranging systems

The authors compare the estimates in passive ranging systems using motion and stereo approaches. It is shown that an integrated approach is necessary to provide better range estimates over a field-of-view (FOV) of interest in helicopter flight. The recursive approach for processing a sequence of stereo images, described together with a recursive motion algorithm (RMA), provides the basis for an integrated method to provide more accurate range information. Results based on motion sequences of stereo images are presented.

Sridhar, Banavar↗

Genome-Wide Association Study Based on Multiple Imputation with Low-Depth Sequencing Data: Application to Biofuel Traits in Reed Canarygrass

Genotyping by sequencing allows for large-scale genetic analyses in plant species with no reference genome, but sets the challenge of sound inference in presence of uncertain genotypes. We report an imputation-based genome-wide association study (GWAS) in reed canarygrass (Phalaris arundinacea L., Phalaris caesia Nees), a cool-season grass species with potential as a biofuel crop. Our study involved two linkage populations and an association panel of 590 reed canarygrass genotypes. Plants were assayed for up to 5228 single nucleotide polymorphism markers and 35 traits. The genotypic markers were derived from lowdepth sequencing with 78% missing data on average. To soundly infer marker-trait associations, multiple imputation (MI) was used: several imputes of the marker data were generated to reflect imputation uncertainty and association tests were performed on marker effects across imputes. A total of nine significant markers were identified, three of which showed significant homology with the Brachypodium dystachion genome. Because no physical map of the reed canarygrass genome was available, imputation was conducted using classification trees. In general, MI showed good consistency with the complete-case analysis and adequate control over imputation uncertainty. A gain in significance of marker effects was achieved through MI, but only for rare cases when missing data were ,45%. In addition to providing insight into the genetic basis of important traits in reed canarygrass, this study presents one of the first applications of MI to genome-wide analyses and provides useful guidelines for conducting GWAS based on genotyping-by-sequencing data.

59 BASIC BIOLOGICAL SCIENCES↗

ADEPT, a dynamic next generation sequencing data error-detection program with trimming

Illumina is the most widely used next generation sequencing technology and produces millions of short reads that contain errors. These sequencing errors constitute a major problem in applications such as de novo genome assembly, metagenomics analysis and single nucleotide polymorphism discovery. In this study, we present ADEPT, a dynamic error detection method, based on the quality scores of each nucleotide and its neighboring nucleotides, together with their positions within the read and compares this to the position-specific quality score distribution of all bases within the sequencing run. This method greatly improves upon other available methods in terms of the true positive rate of error discovery without affecting the false positive rate, particularly within the middle of reads. We conclude that ADEPT is the only tool to date that dynamically assesses errors within reads by comparing position-specific and neighboring base quality scores with the distribution of quality scores for the dataset being analyzed. The result is a method that is less prone to position-dependent under-prediction, which is one of the most prominent issues in error prediction. The outcome is that ADEPT improves upon prior efforts in identifying true errors, primarily within the middle of reads, while reducing the false positive rate.

59 BASIC BIOLOGICAL SCIENCES↗

RNAseq-based transcriptome assembly of Clostridium acetobutylicum for functional genome annotation and discovery

Accurate genome annotations are essential in modern biology and biotechnology, yet they are still largely based on genome sequencing and comparative analyses. We show that the Clostridium acetobutylicum genome annotation can be markedly improved by integrating bioinformatic predictions with RNA sequencing (RNAseq) data. Samples were acquired under butanol, butyrate, and unstressed treatments across various growth conditions. Analysis of an initial assembly revealed errors due to background signals and limitations of assembly algorithms. Hurdles for RNAseq transcriptome mapping include optimizing library complexity and sequencing depth, yet most studies report low sequencing depth and ignore the effect of ribosomal RNA abundance. An integrative analysis was developed to combine motif predictions, single-nucleotide resolution sequencing depth, and library complexity to resolve difficulties in assembly curation. This minimized false positive error and determined gene boundaries, in some cases, to the exact base-pair of prior studies. This will be the first strand-specific transcriptome assembly in a Clostridium organism.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Self-Motion and Depth Estimation from Image Sequences

An image-based version of a computational model of human self-motion perception (developed in collaboration with Dr. Leland S. Stone at NASA Ames Research Center) has been generated and tested. The research included in the grant proposal sought to extend the utility of the self-motion model so that it could be used for explaining and predicting human performance in a greater variety of aerospace applications. The model can now be tested with video input sequences (including computer generated imagery) which enables simulation of human self-motion estimation in a variety of applied settings.

Perrone, John↗

CCD photometry in the globular cluster NGC 288. I - Blue stragglers and main-sequence binary stars

Photometry based on a mosaic of CCD images in B and V is presented for the globular cluster NGC 288. The spatial coverage ranges from the cluster core to about 6 core radii, and stars have been measured over the absolute visual magnitude range -1.2 to 8.4. The cluster is shown to contain a significant number of blue straggler stars in the central regions, and there is an excess of objects brighter and redder than the single-star main-sequence in the color-magnitude plane. These objects are interpreted as a population of main-sequence binary stars. With this interpretation, the explicity measured fraction of binary stars is 10 percent, which sets a lower limit for the total binary population.

Bolte, Michael↗

Recruiting Human Microbiome Shotgun Data to Site-Specific Reference Genomes

The human body consists of innumerable multifaceted environments that predispose colonization by a number of distinct microbial communities, which play fundamental roles in human health and disease. In addition to community surveys and shotgun metagenomes that seek to explore the composition and diversity of these microbiomes, there are significant efforts to sequence reference microbial genomes from many body sites of healthy adults. To illustrate the utility of reference genomes when studying more complex metagenomes, we present a reference-based analysis of sequence reads generated from 55 shotgun metagenomes, selected from 5 major body sites, including 16 sub-sites. Interestingly, between 13% and 92% (62.3% average) of these shotgun reads were aligned to a then-complete list of 2780 reference genomes, including 1583 references for the human microbiome. However, no reference genome was universally found in all body sites. For any given metagenome, the body site-specific reference genomes, derived from the same body site as the sample, accounted for an average of 58.8% of the mapped reads. While different body sites did differ in abundant genera, proximal or symmetrical body sites were found to be most similar to one another. The extent of variation observed, both between individuals sampled within the same microenvironment, or at the same site within the same individual over time, calls into question comparative studies across individuals even if sampled at the same body site. This study illustrates the high utility of reference genomes and the need for further site-specific reference microbial genome sequencing, even within the already well-sampled human microbiome.

59 BASIC BIOLOGICAL SCIENCES↗

Integration of motion and stereo sensors in passive ranging systems

A recursive approach is described for processing a sequence of stereo images. It will be the basis for an integrated stereo and motion method to provide more accurate range information using a passive ranging system. Results based on motion sequences of stereo images are presented. The approach is also applicable to other autonomous systems and in robotics.

Sridhar, Banavar↗

Tissue-specific differences in metabolites and transcripts contribute to the heterogeneity of ricinoleic acid accumulation in Ricinus communis L. (castor) seeds

Castor ( Ricinus communis L.) seeds are valued for their production of oils which can comprise up to 90% hydroxy-fatty acids (ricinoleic acid). Castor oil contains mono-, di- and tri- ricinoleic acid containing triacylglycerols (TAGs). Although the enzymatic synthesis of ricinoleic acid is well described, the differential compartmentalization of these TAG molecular species has remained undefined. To examine the distribution of hydroxy fatty acid accumulation within the endosperm and embryo tissues of castor seeds. Matrix assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) was used to map the distribution of triacylglycerols in tissue sections of castor seeds. In addition, the endosperm and embryo (cotyledons and embryonic axis) tissues were dissected and extracted for quantitative lipidomics analysis and Illumina-based RNA deep sequencing. Results: This study revealed an unexpected heterogeneous tissue distribution of mono-, di- and tri- hydroxy-triacylglycerols in the embryo and endosperm tissues of castor seeds. Pathway analysis based on transcript abundance suggested that distinct embryo- and endosperm-specific mechanisms may exist for the shuttling of ricinoleic acid away from phosphatidylcholine (PC) and into hydroxy TAG production. The embryo-biased mechanism appears to favor removal of ricinoleic acid from PC through phophatidylcholine: diacylglycerol acylytransferase (PDAT) while the endosperm pathway appears to remove ricinoleic acid from the PC pool by preferences of phospholipase A (PLA 2 α) and/or phosphatidylcholine: diacylglycerol cholinepshphotransferase PDCT. In conclusion, collectively, a combination of lipidomics and transcriptomics analyses revealed previously undefined spatial aspects of hydroxy fatty acid metabolism in castor seeds. These studies underscore a need for tissue-specific studies as a means to better understand the regulation of triacylglycerol accumulation in oilseeds.

59 BASIC BIOLOGICAL SCIENCES↗

Reasoning about real-time systems with temporal interval logic constraints on multi-state automata

Models of real-time systems using a single paradigm often turn out to be inadequate, whether the paradigm is based on states, rules, event sequences, or logic. A model-based approach to reasoning about real-time systems is presented in which a temporal interval logic called TIL is employed to define constraints on a new type of high level automata. The combination, called hierarchical multi-state (HMS) machines, can be used to model formally a real-time system, a dynamic set of requirements, the environment, heuristic knowledge about planning-related problem solving, and the computational states of the reasoning mechanism. In this framework, mathematical techniques were developed for: (1) proving the correctness of a representation; (2) planning of concurrent tasks to achieve goals; and (3) scheduling of plans to satisfy complex temporal constraints. HMS machines allow reasoning about a real-time system from a model of how truth arises instead of merely depending of what is true in a system.

Gabrielian, Armen↗

Knowledge-based decision support for Space Station assembly sequence planning

A complete Personal Analysis Assistant (PAA) for Space Station Freedom (SSF) assembly sequence planning consists of three software components: the system infrastructure, intra-flight value added, and inter-flight value added. The system infrastructure is the substrate on which software elements providing inter-flight and intra-flight value-added functionality are built. It provides the capability for building representations of assembly sequence plans and specification of constraints and analysis options. Intra-flight value-added provides functionality that will, given the manifest for each flight, define cargo elements, place them in the National Space Transportation System (NSTS) cargo bay, compute performance measure values, and identify violated constraints. Inter-flight value-added provides functionality that will, given major milestone dates and capability requirements, determine the number and dates of required flights and develop a manifest for each flight. The current project is Phase 1 of a projected two phase program and delivers the system infrastructure. Intra- and inter-flight value-added were to be developed in Phase 2, which has not been funded. Based on experience derived from hundreds of projects conducted over the past seven years, ISX developed an Intelligent Systems Engineering (ISE) methodology that combines the methods of systems engineering and knowledge engineering to meet the special systems development requirements posed by intelligent systems, systems that blend artificial intelligence and other advanced technologies with more conventional computing technologies. The ISE methodology defines a phased program process that begins with an application assessment designed to provide a preliminary determination of the relative technical risks and payoffs associated with a potential application, and then moves through requirements analysis, system design, and development.

Source record↗

G4Boost: a machine learning-based tool for quadruplex identification and stability prediction

Background: G-quadruplexes (G4s), formed within guanine-rich nucleic acids, are secondary structures involved in important biological processes. Although every G4 motif has the potential to form a stable G4 structure, not every G4 motif would, and accurate energy-based methods are needed to assess their structural stability. Here, we present a decision tree-based prediction tool, G4Boost, to identify G4 motifs and predict their secondary structure folding probability and thermodynamic stability based on their sequences, nucleotide compositions, and estimated structural topologies. Results: G4Boost predicted the quadruplex folding state with an accuracy greater then 93% and an F1-score of 0.96, and the folding energy with an RMSE of 4.28 and R 2 of 0.95 only by the means of sequence intrinsic feature. G4Boost was successfully applied and validated to predict the stability of experimentally-determined G4 structures, including for plants and humans. Conclusion: G4Boost outperformed the three machine-learning based prediction tools, DeepG4, Quadron, and G4RNA Screener, in terms of both accuracy and F1-score, and can be highly useful for G4 prediction to understand gene regulation across species including plants and humans.

59 BASIC BIOLOGICAL SCIENCES↗

Correlation of Advanced Accelerated Stress Testing with Polyamide-Based Photovoltaic Backsheet Field-Failures

Cracking of polyamide (PA)-based photovoltaic (PV) backsheet materials has been widely reported for field-aged modules. Failure was not detected by conventional accelerated stress tests (ASTs), which lacked the necessary combination of stress factors and/or factor sequences. PA-based AAA backsheet cracking has since been reproduced through combined and sequential stress testing. Planar- and cross-sectional-optical microscopy as well as Fourier-transform Infrared Spectroscopy (FTIR), have been used to elucidate the mechanical and chemical changes which lead to failure of the backsheet. Field-aged backsheet samples demonstrating failure in various climates (including locations in China and Italy) are also analyzed. Through the analysis, a comparison is made between the different stress testing protocols and the field-aged samples to validate relevance of the advanced stress tests. It is shown that the changes induced through combined-accelerated stress testing (C-AST) were most representative of changes induced by the field, supporting the relevance of C-AST and providing validity for the test protocol.

14 SOLAR ENERGY↗

Methods and systems for detecting cancer

A technique for classifying lesions as malignant or benign is disclosed. The technique can include: cooling an area of skin including a lesion of a patient to initiate a warm-up process; receiving a temporal sequence of thermal images of the area of skin representing a thermal recovery of the area of skin, the temporal sequence of thermal images generated by an infrared camera; generating a temporal profile of the thermal recovery based on the temporal sequence of thermal images; analyzing temporal statistical properties of the temporal profile; determining a malignancy probability that the lesion is malignant based on an analysis of the temporal profile, wherein the determining includes extracting one or more statistical features based on continuous-time stochastic signals in the sequence of thermal images; and classifying the lesion based on the malignancy probability.

47 OTHER INSTRUMENTATION↗