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

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At least 1,351 records · Page 75
All sourcesNASA NTRSDOE OSTI
DOE OSTI2015

SU-E-J-257: A PCA Model to Predict Adaptive Changes for Head&neck Patients Based On Extraction of Geometric Features From Daily CBCT Datasets

Purpose: Using daily cone beam CTs (CBCTs) to develop principal component analysis (PCA) models of anatomical changes in head and neck (H&N) patients and to assess the possibility of using these prospectively in adaptive radiation therapy (ART). Methods: Planning CT (pCT) images of 4 H&N patients were deformed to model several different systematic changes in patient anatomy during the course of the radiation therapy (RT). A Pinnacle plugin was used to linearly interpolate the systematic change in patient for the 35 fraction RT course and to generate a set of 35 synthetic CBCTs. Each synthetic CBCT represents the systematic change in patient anatomy for each fraction. Deformation vector fields (DVFs) were acquired between the pCT and synthetic CBCTs with random fraction-to-fraction changes were superimposed on the DVFs. A patient-specific PCA model was built using these DVFs containing systematic plus random changes. It was hypothesized that resulting eigenDVFs (EDVFs) with largest eigenvalues represent the major anatomical deformations during the course of treatment. Results: For all 4 patients, the PCA model provided different results depending on the type and size of systematic change in patient’s body. PCA was more successful in capturing the systematic changes early in the treatment course when these were of a larger scale with respect to the random fraction-to-fraction changes in patient’s anatomy. For smaller scale systematic changes, random changes in patient could completely “hide” the systematic change. Conclusion: The leading EDVF from the patientspecific PCA models could tentatively be identified as a major systematic change during treatment if the systematic change is large enough with respect to random fraction-to-fraction changes. Otherwise, leading EDVF could not represent systematic changes reliably. This work is expected to facilitate development of population-based PCA models that can be used to prospectively identify significant anatomical changes early in treatment. This work is supported in part by a grant from Varian Medical Systems, Palo Alto, CA.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2015

SU-E-T-753: Three-Dimensional Dose Distributions of Incident Proton Particle in the Polymer Gel Dosimeter and the Radiochromic Gel Dosimeter: A Simulation Study with MCNP Code

Purpose: The purpose of this study is to estimate the three-dimensional dose distributions in the polymer and the radiochromic gel dosimeter, and to identify the detectability of both gel dosimeters by comparing with the water phantom in case of irradiating the proton particles. Methods: The normoxic polymer gel and the LCV micelle radiochromic gel were used in this study. The densities of polymer and the radiochromic gel dosimeter were 1.024 and 1.005 g/cm{sup 3}, respectively. The dose distributions of protons in the polymer and radiochromic gel were simulated using Monte Carlo radiation transport code (MCNPX, Los Alamos National Laboratory). The shape of phantom irradiated by proton particles was a hexahedron with the dimension of 12.4 × 12.4 × 15.0 cm{sup 3}. The energies of proton beam were 50, 80, and 140 MeV energies were directed to top of the surface of phantom. The cross-sectional view of proton dose distribution in both gel dosimeters was estimated with the water phantom and evaluated by the gamma evaluation method. In addition, the absorbed dose(Gy) was also calculated for evaluating the proton detectability. Results: The evaluation results show that dose distributions in both gel dosimeters at intermediated section and Bragg-peak region are similar with that of the water phantom. At entrance section, however, inconsistencies of dose distribution are represented, compared with water. The relative absorbed doses in radiochromic and polymer gel dosimeter were represented to be 0.47 % and 2.26 % difference, respectively. These results show that the radiochromic gel dosimeter was better matched than the water phantom in the absorbed dose evaluation. Conclusion: The polymer and the radiochromic gel dosimeter show similar characteristics in dose distributions for the proton beams at intermediate section and Bragg-peak region. Moreover the calculated absorbed dose in both gel dosimeters represents similar tendency by comparing with that in water phantom.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2015

SU-E-T-782: Using Light Output From Doped Plastic Scintillators to Resolve the Linear Energy Transfer Spectrum of Clinical Electron Beams

Purpose: This research seeks to develop a portable, clinically-suitable linear energy transfer (LET) detector. In radiotherapy, absorbed dose is commonly used to measure the amount of delivered radiation, though, it is not a good indicator of actual biological damage. LET is the energy absorbed per unit length by a medium along charged particle’s pathway; studies have shown that LET correlates well with relative biological effectiveness (RBE). Methods: According to Birks’ law, light output of plastic scintillators is stopping-power dependent. This dependency can be varied through doping by various high-Z elements. By measuring light output signals of differently doped plastic scintillators (represented by column vector S, where each row corresponds to different scintillator material), the fluence of charged particles of a given LET (represented by column vector Φ, where each row corresponds to different LET bins) can be unfolded by S=R*Φ where R is system response matrix (each row represents a different scintillator, each column corresponds to different electron LET). Monte Carlo (MC) GEANT4.10.1 was used to evaluate ideal detector response of BC408 scintillating material doped with various concentrations of several high Z dopants. Measurements were performed to validate MC. Results: Signal for 1%-lead doped BC408 and the non-doped scintillator was measured experimentally by guiding light emitted by the scintillator (via in-house made taper, fiber system) to a PMT and then an electrometer. Simulations of 1%Pb-doped scintillator to non-doped scintillator revealed 9.3% reduction in light output for 6 MeV electrons which compared well (within uncertainty) with measurements showing 10% reduction (6MeV electrons). Conclusion: Measurements were used to validate MC simulation of light output from doped scintillators. The doping of scintillators is a viable technique to induce LET dependence. Our goal is to use this effect to resolve the LET spectrum of an incident beam.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2015

Spectrotemporal CT data acquisition and reconstruction at low dose

Purpose: X-ray computed tomography (CT) is widely used, both clinically and preclinically, for fast, high-resolution anatomic imaging; however, compelling opportunities exist to expand its use in functional imaging applications. For instance, spectral information combined with nanoparticle contrast agents enables quantification of tissue perfusion levels, while temporal information details cardiac and respiratory dynamics. The authors propose and demonstrate a projection acquisition and reconstruction strategy for 5D CT (3D + dual energy + time) which recovers spectral and temporal information without substantially increasing radiation dose or sampling time relative to anatomic imaging protocols. Methods: The authors approach the 5D reconstruction problem within the framework of low-rank and sparse matrix decomposition. Unlike previous work on rank-sparsity constrained CT reconstruction, the authors establish an explicit rank-sparse signal model to describe the spectral and temporal dimensions. The spectral dimension is represented as a well-sampled time and energy averaged image plus regularly undersampled principal components describing the spectral contrast. The temporal dimension is represented as the same time and energy averaged reconstruction plus contiguous, spatially sparse, and irregularly sampled temporal contrast images. Using a nonlinear, image domain filtration approach, the authors refer to as rank-sparse kernel regression, the authors transfer image structure from the well-sampled time and energy averaged reconstruction to the spectral and temporal contrast images. This regularization strategy strictly constrains the reconstruction problem while approximately separating the temporal and spectral dimensions. Separability results in a highly compressed representation for the 5D data in which projections are shared between the temporal and spectral reconstruction subproblems, enabling substantial undersampling. The authors solved the 5D reconstruction problem using the split Bregman method and GPU-based implementations of backprojection, reprojection, and kernel regression. Using a preclinical mouse model, the authors apply the proposed algorithm to study myocardial injury following radiation treatment of breast cancer. Results: Quantitative 5D simulations are performed using the MOBY mouse phantom. Twenty data sets (ten cardiac phases, two energies) are reconstructed with 88 μm, isotropic voxels from 450 total projections acquired over a single 360° rotation. In vivo 5D myocardial injury data sets acquired in two mice injected with gold and iodine nanoparticles are also reconstructed with 20 data sets per mouse using the same acquisition parameters (dose: ∼60 mGy). For both the simulations and the in vivo data, the reconstruction quality is sufficient to perform material decomposition into gold and iodine maps to localize the extent of myocardial injury (gold accumulation) and to measure cardiac functional metrics (vascular iodine). Their 5D CT imaging protocol represents a 95% reduction in radiation dose per cardiac phase and energy and a 40-fold decrease in projection sampling time relative to their standard imaging protocol. Conclusions: Their 5D CT data acquisition and reconstruction protocol efficiently exploits the rank-sparse nature of spectral and temporal CT data to provide high-fidelity reconstruction results without increased radiation dose or sampling time.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2016

Bone-marrow densitometry: Assessment of marrow space of human vertebrae by single energy high resolution-quantitative computed tomography

Purpose: Accurate noninvasive assessment of vertebral bone marrow fat fraction is important for diagnostic assessment of a variety of disorders and therapies known to affect marrow composition. Moreover, it provides a means to correct fat-induced bias of single energy quantitative computed tomography (QCT) based bone mineral density (BMD) measurements. The authors developed new segmentation and calibration methods to obtain quantitative surrogate measures of marrow-fat density in the axial skeleton. Methods: The authors developed and tested two high resolution-QCT (HR-QCT) based methods which permit segmentation of bone voids in between trabeculae hypothesizing that they are representative of bone marrow space. The methods permit calculation of marrow content in units of mineral equivalent marrow density (MeMD). The first method is based on global thresholding and peeling (GTP) to define a volume of interest away from the transition between trabecular bone and marrow. The second method, morphological filtering (MF), uses spherical elements of different radii (0.1–1.2 mm) and automatically places them in between trabeculae to identify regions with large trabecular interspace, the bone-void space. To determine their performance, data were compared ex vivo to high-resolution peripheral CT (HR-pQCT) images as the gold-standard. The performance of the methods was tested on a set of excised human vertebrae with intact bone marrow tissue representative of an elderly population with low BMD. Results: 86% (GTP) and 87% (MF) of the voxels identified as true marrow space on HR-pQCT images were correctly identified on HR-QCT images and thus these volumes of interest can be considered to be representative of true marrow space. Within this volume, MeMD was estimated with residual errors of 4.8 mg/cm{sup 3} corresponding to accuracy errors in fat fraction on the order of 5% both for GTP and MF methods. Conclusions: The GTP and MF methods on HR-QCT images permit noninvasive localization and densitometric assessment of marrow fat with residual accuracy errors sufficient to study disorders and therapies known to affect bone marrow composition. Additionally, the methods can be used to correct BMD for fat induced bias. Application and testing in vivo and in longitudinal studies are warranted to determine the clinical performance and value of these methods.

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗
DOE OSTI2016

SU-D-BRB-05: Quantum Learning for Knowledge-Based Response-Adaptive Radiotherapy

Purpose: There is tremendous excitement in radiotherapy about applying data-driven methods to develop personalized clinical decisions for real-time response-based adaptation. However, classical statistical learning methods lack in terms of efficiency and ability to predict outcomes under conditions of uncertainty and incomplete information. Therefore, we are investigating physics-inspired machine learning approaches by utilizing quantum principles for developing a robust framework to dynamically adapt treatments to individual patient’s characteristics and optimize outcomes. Methods: We studied 88 liver SBRT patients with 35 on non-adaptive and 53 on adaptive protocols. Adaptation was based on liver function using a split-course of 3+2 fractions with a month break. The radiotherapy environment was modeled as a Markov decision process (MDP) of baseline and one month into treatment states. The patient environment was modeled by a 5-variable state represented by patient’s clinical and dosimetric covariates. For comparison of classical and quantum learning methods, decision-making to adapt at one month was considered. The MDP objective was defined by the complication-free tumor control (P{sup +}=TCPx(1-NTCP)). A simple regression model represented state-action mapping. Single bit in classical MDP and a qubit of 2-superimposed states in quantum MDP represented the decision actions. Classical decision selection was done using reinforcement Q-learning and quantum searching was performed using Grover’s algorithm, which applies uniform superposition over possible states and yields quadratic speed-up. Results: Classical/quantum MDPs suggested adaptation (probability amplitude ≥0.5) 79% of the time for splitcourses and 100% for continuous-courses. However, the classical MDP had an average adaptation probability of 0.5±0.22 while the quantum algorithm reached 0.76±0.28. In cases where adaptation failed, classical MDP yielded 0.31±0.26 average amplitude while the quantum approach averaged a more optimistic 0.57±0.4, but with high phase fluctuations. Conclusion: Our results demonstrate that quantum machine learning approaches provide a feasible and promising framework for real-time and sequential clinical decision-making in adaptive radiotherapy.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2016

SU-F-I-37: How Fat Distribution and Table Height Affect Estimates of Patient Size in CT Scanning: A Phantom Study

Purpose: Localizer projection radiographs acquired prior to CT scans are used to estimate patient size, affecting the function of Automatic Tube Current Modulation (ATCM) and hence CTDIvol and SSDE. Due to geometric effects, the projected patient size varies with scanner table height and with the orientation of the localizer (AP versus PA). This study sought to determine if patient size estimates made from localizer scans is affected by variations in fat distribution, specifically when the widest part of the patient is not at the geometric center of the patient. Methods: Lipid gel bolus material was wrapped around an anthropomorphic phantom to simulate two different body mass distributions. The first represented a patient with fairly rigid fat and had a generally oval shape. The second was bell-shaped, representing corpulent patients more susceptible to gravity’s lustful tug. Each phantom configuration was imaged using an AP localizer and then a PA localizer. This was repeated at various scanner table heights. The width of the phantom was measured from the localizer and diagnostic images using in-house software. Results: 1) The projected phantom width varied up to 39% as table height changed.2) At some table heights, the width of the phantom, designed to represent larger patients, exceeded the localizer field of view, resulting in an underestimation of the phantom width.3) The oval-shaped phantom approached a normalized phantom width of 1 at a table height several centimeters lower (AP localizer) or higher (PA localizer) than did the bell-shaped phantom. Conclusion: Accurate estimation of patient size from localizer scans is dependent on patient positioning with respect to scanner isocenter and is limited in large patients. Further, patient size is more accurately measured on projection images if the widest part of the patient, rather than the geometric center of the patient, is positioned at scanner isocenter.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2016

SU-F-T-374: Dosimetric Effects of Irradiation Through a Bilateral Hip Prosthesis in a MRI Linac

Purpose: To evaluate the interface effects when irradiating through a hip prosthesis in the presence of an orthogonal 1.5 T magnetic field using Monte Carlo simulations. Methods: A 20×20×38 cm virtual phantom with two 5×5×5 cm sections of bilateral titanium hip prosthesis was created in GPU-based Monte Carlo (MC) algorithm (GPUMCD, Elekta AB, Stockholm Sweden). The lateral prosthesis spacing was based on a representative patient CT scan. A treatment SAD of 143.5 cm was chosen, corresponding to the Elekta AB MRI Linac and the beam energy distribution was sampled from a histogram representing the true MRI Linac spectrum. A magnetic field of 1.5 T was applied perpendicular to the plane of irradiation. Dose was calculated, in voxels of side 1 mm, for 2×2, 5×5, and 10×10 cm treatment field sizes with normal beam incidence (gantry at 90° or 270°) and at 5° and 10° from normal, representing the range of incidence through the bilateral prosthesis. Results: With magnetic field ON (B-On) and normal beam incidence the backscatter dose at the interfaces of proximal and distal implants is reduced for all the field sizes compared to the magnetic field OFF (B-Off) case. The absolute reduction in doses at the interface was in the range of 12.93% to 13.16% for the proximal implant and 13.57% to 16.12% for the distal implant. Similarly for the oblique incidences of 5o and 10o the dose in the plane adjacent to the prosthetic implants is lower when the magnetic field is ON. Conclusion: The dosimetric effects of irradiating through a hip prosthesis in the presence of a transverse magnetic field have been determined using MC simulation. The backscatter dose reduction translates into significantly lower hot spots at the prosthetic interfaces, which are otherwise substantially high in the absence of the magnetic field. This project was supported through funding provided by ElektaTM.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2016

SU-G-IeP1-12: Size Selective Arterial Cerebral Blood Volume Mapping Using Multiple Inversion Time Arterial Spin Labeling

Purpose: To demonstrate the feasibility of a novel method for size specific arterial cerebral blood volume (aCBV) mapping using pseudo-continuous arterial spin labeling (PCASL), with multiple TI. Methods: Multiple PCASL images were obtained from a subject with TI of [300, 400, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000] ms. Each TI pair was averaged six times. Two scans were performed: one without a flow crusher gradient and the other with a crusher gradient (10cm/s in three directions) to remove signals from large arteries. Scan times were 5min. without a crusher gradient and 5.5 min with a crusher gradient. Non-linear fitting algorithm finds the minimum mean squared solution of per-voxel based aCBV, cerebral blood flow, and arterial transit time, and fits the data into a hemodynamic model that represents superposition of blood volume and flow components within a single voxel. Results: aCBV maps with a crusher gradient represent signals from medium and small sized arteries, while those without a crusher gradient represent signals from all sized arteries, indicating that flow crusher gradients can be effectively employed to achieve size-specific aCBV mapping. Regardless of flow crusher, the CBF and ATT maps are very similar in appearance. Conclusion: Quantitative size selective blood volume mapping controlled by a flow crusher is feasible without additional information because the ASL quantification process doesn’t require an arterial input function measured from a large artery. The size specific blood volume mapping is not interfered by sSignals from large arteries do not interfere with size specific aCBV mapping in the applications of interest in for applications in which only medium or small arteries are of interest.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2016

SU-G-JeP4-08: External and Internal Motion Assessments with Amplitude and Time Based Reconstruction and AveCT

Purpose: To investigate the effects of external surrogate and tumour motion by observing the reconstructed phases and AveCT in an Amplitude and Time based 4DCT. Methods: Based on patient motion studies, Cos6 and sinusoidal motions were simulated as external surrogate and tumour motions in a motion phantom. The diaphragm and tumour motions may or may not display the same waveform therefore the same and different waveforms were programmed into the phantom, scanned and reconstructed based on Amplitude and Time. The AveCT and phases were investigated with these different scenarios. The AveCT phantom images were also compared with CBCT phantom images programmed with the same motions. Results: For the same surrogate and tumour sin motions, the phases (Amplitude and Time) and AveCT indicated similar motions based on the position of the BB at the slice and displayed contrast values respectively. For cos6 motions, due to the varied time the tumour spends at each position, the Amplitude and Time based phases differed. The AveCT images represented the actual tumour motions and the Time and Amplitude based phases were represented by the surrogate with varied times. Conclusion: Different external surrogate and tumour motions may result in different displayed image motions when observing the AveCT and reconstructed phases. During the 4DCT, the surrogate motion is readily available for observation of the amplitude and time of the diaphragm position. Following image reconstruction, the user may need to observe the AveCT in addition to the reconstructed phases to comprehend the time weightings of the tumour motion during the scan. This may also apply to 3D CBCT images where the displayed tumour position in the images is influenced by the long duration of the CBCT. Knowledge of the tumour motion represented by the greyscale of the AveCT may also assist in CBCT treatment beam verification matching.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2016

TU-AB-BRC-05: Creation of a Monte Carlo TrueBeam Model by Reproducing Varian Phase Space Data

Purpose: To create a Varian TrueBeam 6 MV FFF Monte Carlo model using BEAMnrc/EGSnrc that accurately reproduces the Varian representative dataset, followed by tuning the model’s source parameters to accurately reproduce in-house measurements. Methods: A BEAMnrc TrueBeam model for 6 MV FFF has been created by modifying a validated 6 MV Varian CL21EX model. Geometric dimensions and materials were adjusted in a trial and error approach to match the fluence and spectra of TrueBeam phase spaces output by the Varian VirtuaLinac. Once the model’s phase space matched Varian’s counterpart using the default source parameters, it was validated to match 10 × 10 cm{sup 2} Varian representative data obtained with the IBA CC13. The source parameters were then tuned to match in-house 5 × 5 cm{sup 2} PTW microDiamond measurements. All dose to water simulations included detector models to include the effects of volume averaging and the non-water equivalence of the chamber materials, allowing for more accurate source parameter selection. Results: The Varian phase space spectra and fluence were matched with excellent agreement. The in-house model’s PDD agreement with CC13 TrueBeam representative data was within 0.9% local percent difference beyond the first 3 mm. Profile agreement at 10 cm depth was within 0.9% local percent difference and 1.3 mm distance-to-agreement in the central axis and penumbra regions, respectively. Once the source parameters were tuned, PDD agreement with microDiamond measurements was within 0.9% local percent difference beyond 2 mm. The microDiamond profile agreement at 10 cm depth was within 0.6% local percent difference and 0.4 mm distance-to-agreement in the central axis and penumbra regions, respectively. Conclusion: An accurate in-house Monte Carlo model of the Varian TrueBeam was achieved independently of the Varian phase space solution and was tuned to in-house measurements. KO acknowledges partial support by the CREATE Medical Physics Research Training Network grant of the Natural Sciences and Engineering Research Council (Grant number: 432290).

60 APPLIED LIFE SCIENCES↗
DOE OSTI2016

TH-AB-207A-05: A Fully-Automated Pipeline for Generating CT Images Across a Range of Doses and Reconstruction Methods

Purpose: To evaluate the robustness of CAD or Quantitative Imaging methods, they should be tested on a variety of cases and under a variety of image acquisition and reconstruction conditions that represent the heterogeneity encountered in clinical practice. The purpose of this work was to develop a fully-automated pipeline for generating CT images that represent a wide range of dose and reconstruction conditions. Methods: The pipeline consists of three main modules: reduced-dose simulation, image reconstruction, and quantitative analysis. The first two modules of the pipeline can be operated in a completely automated fashion, using configuration files and running the modules in a batch queue. The input to the pipeline is raw projection CT data; this data is used to simulate different levels of dose reduction using a previously-published algorithm. Filtered-backprojection reconstructions are then performed using FreeCT-wFBP, a freely-available reconstruction software for helical CT. We also added support for an in-house, model-based iterative reconstruction algorithm using iterative coordinate-descent optimization, which may be run in tandem with the more conventional recon methods. The reduced-dose simulations and image reconstructions are controlled automatically by a single script, and they can be run in parallel on our research cluster. The pipeline was tested on phantom and lung screening datasets from a clinical scanner (Definition AS, Siemens Healthcare). Results: The images generated from our test datasets appeared to represent a realistic range of acquisition and reconstruction conditions that we would expect to find clinically. The time to generate images was approximately 30 minutes per dose/reconstruction combination on a hybrid CPU/GPU architecture. Conclusion: The automated research pipeline promises to be a useful tool for either training or evaluating performance of quantitative imaging software such as classifiers and CAD algorithms across the range of acquisition and reconstruction parameters present in the clinical environment. Funding support: NIH U01 CA181156; Disclosures (McNitt-Gray): Institutional research agreement, Siemens Healthcare; Past recipient, research grant support, Siemens Healthcare; Consultant, Toshiba America Medical Systems; Consultant, Samsung Electronics.

60 APPLIED LIFE SCIENCES↗
DOE OSTI2018

Diversity of Active Viral Infections within the Sphagnum Microbiome

Sphagnum -dominated peatlands play an important role in global carbon storage and represent significant sources of economic and ecological value. While recent efforts to describe microbial diversity and metabolic potential of the Sphagnum microbiome have demonstrated the importance of its microbial community, little is known about the viral constituents. We used metatranscriptomics to describe the diversity and activity of viruses infecting microbes within the Sphagnum peat bog. The vegetative portions of six Sphagnum plants were obtained from a peatland in northern Minnesota, and the total RNA was extracted and sequenced. Metatranscriptomes were assembled and contigs were screened for the presence of conserved virus marker genes. Using bacteriophage capsid protein gp23 as a marker for phage diversity, we identified 33 contigs representing undocumented phages that were active in the community at the time of sampling. Similarly, RNA-dependent RNA polymerase and the nucleocytoplasmic large DNA virus (NCLDV) major capsid protein were used as markers for single-stranded RNA (ssRNA) viruses and NCLDV, respectively. In total, 114 contigs were identified as originating from undescribed ssRNA viruses, 22 of which represent nearly complete genomes. An additional 64 contigs were identified as being from NCLDVs. Finally, 7 contigs were identified as putative virophage or polinton-like viruses. We developed co-occurrence networks with these markers in relation to the expression of potential-host housekeeping gene rpb1 to predict virus-host relationships, identifying 13 groups. Together, our approach offers new tools for the identification of virus diversity and interactions in understudied clades and suggests that viruses may play a considerable role in the ecology of the Sphagnum microbiome. Sphagnum -dominated peatlands play an important role in maintaining atmospheric carbon dioxide levels by modifying conditions in the surrounding soil to favor the growth of over that of other plant species. This lowers the rate of decomposition and facilitates the accumulation of fixed carbon in the form of partially decomposed biomass. The unique environment produced by Sphagnum enriches for the growth of a diverse microbial consortia that benefit from and support the moss's growth, while also maintaining the hostile soil conditions. While a growing body of research has begun to characterize the microbial groups that colonize Sphagnum , little is currently known about the ecological factors that constrain community structure and define ecosystem function. Top-down population control by viruses is almost completely undescribed. Furthermore, this study provides insight into the significant viral influence on the Sphagnum microbiome and identifies new potential model systems to study virus-host interactions in the peatland ecosystem.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2017

Divergence in Gene Regulation Contributes to Sympatric Speciation of Shewanella baltica Strains

Niche partitioning and sequence evolution drive genomic and phenotypic divergence, which ultimately leads to bacterial diversification. This study investigated the genomic composition of two Shewanella baltica clades previously identified through multilocus sequencing typing and recovered from the redox transition zone in the central Baltic Sea. Comparative genomic analysis revealed significantly higher interclade than intraclade genomic dissimilarity and that a subset of genes present in clade A were associated with potential adaptation to respiration of sulfur compounds present in the redox transition zone. The transcriptomic divergence between two representative strains of clades A and D, OS185 and OS195, was also characterized and revealed marked regulatory differences. We found that both the transcriptional divergence of shared genes and expression of strain-specific genes led to differences in regulatory patterns between strains that correlate with environmental redox niches. For example, under anoxic conditions of respiratory nitrate ammonification, OS185—the strain isolated from a nitrate-rich environment—upregulated nearly twice the number of shared genes upregulated by OS195—the strain isolated from an H 2 S-containing anoxic environment. On the other hand, OS195 showed stronger induction of strain-specific genes, especially those associated with sulfur compound respiration, under thiosulfate-reducing conditions. A positive association between the level of transcriptional divergence and the level of sequence divergence for shared genes was also noted. Our results provide further support for the hypothesis that genomic changes impacting transcriptional regulation play an important role in the diversification of ecologically distinct populations. IMPORTANCE: This study examined potential mechanisms through which co-occurring Shewanella baltica strains diversified to form ecologically distinct populations. At the time of isolation, the strains studied composed the major fraction of culturable nitrate-reducing communities in the Baltica Sea. Analysis of genomic content of 13 S. baltica strains from two clades representing different ecotypes demonstrated that one clade specifically possesses a number of genes that could favor successful adaptation to respire sulfur compounds in the portion of the water column from which these strains were isolated. In addition, transcriptional profiling of fully sequenced strains representative of these two clades, OS185 and OS195, under oxygen-, nitrate-, and thiosulfate-respiring conditions demonstrated that the strains exhibit relatively similar transcriptional responses during aerobic growth but more-distinct transcriptional responses under nitrate- and thiosulfate-respiring conditions. Results from this study provide insights into how genomic and gene regulatory diversification together impacted the redox specialization of the S. baltica strains.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2022

Community Structure and Microbial Associations in Sediment-Free Methanotrophic Enrichment Cultures from a Marine Methane Seep

We report Syntrophic consortia of anaerobic methanotrophic archaea (ANME) and sulfate-reducing bacteria (SRB) consume large amounts of methane and serve as the foundational microorganisms in marine methane seeps. Despite their importance in the carbon cycle, research on the physiology of ANME-SRB consortia has been hampered by the slow growth and complex physicochemical environment the consortia inhabit. Here, we report successful sediment-free enrichment of ANME-SRB consortia from deep-sea methane seep sediments in the Santa Monica Basin, California. Anoxic Percoll density gradients and size-selective filtration were used to separate ANME-SRB consortia from sediment particles and single cells to accelerate the cultivation process. Over a 3-year period, a subset of the sediment-associated ANME and SRB lineages, predominantly comprised of ANME-2a/2b (“Candidatus Methanocomedenaceae”) and their syntrophic bacterial partners, SEEP-SRB1/2, adapted and grew under defined laboratory conditions. Metagenome-assembled genomes from several enrichments revealed that ANME-2a, SEEP-SRB1, and Methanococcoides in different enrichments from the same inoculum represented distinct species, whereas other coenriched microorganisms were closely related at the species level. This suggests that ANME, SRB, and Methanococcoides are more genetically diverse than other members in methane seeps. Flow cytometry sorting and sequencing of cell aggregates revealed that Methanococcoides, Anaerolineales, and SEEP-SRB1 were overrepresented in multiple ANME-2a cell aggregates relative to the bulk metagenomes, suggesting they were physically associated and possibly interacting. Overall, this study represents a successful case of selective cultivation of anaerobic slow-growing microorganisms from sediments based on their physical characteristics, introducing new opportunities for detailed genomic, physiological, biochemical, and ecological analyses. Biological anaerobic oxidation of methane (AOM) coupled with sulfate reduction represents a large methane sink in global ocean sediments. Methane consumption is carried out by syntrophic archaeal-bacterial consortia and fuels a unique ecosystem, yet the interactions in these slow-growing syntrophic consortia and with other associated community members remain poorly understood. The significance of this study is the establishment of sediment-free enrichment cultures of anaerobic methanotrophic archaea and sulfate-reducing bacteria performing AOM with sulfate using selective cultivation approaches based on size, density, and metabolism. By reconstructing microbial genomes and analyzing community composition of the enrichment cultures and cell aggregates, we shed light on the diversity of microorganisms physically associated with AOM consortia beyond the core syntrophic partners. These enrichment cultures offer simplified model systems to extend our understanding of the diversity of microbial interactions within marine methane seeps.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2017

Panels of HIV-1 Subtype C Env Reference Strains for Standardized Neutralization Assessments

In the search for effective immunologic interventions to prevent and treat HIV-1 infection, standardized reference reagents are a cost-effective way to maintain robustness and reproducibility among immunological assays. To support planned and ongoing studies where clade C predominates, here we describe three virus panels, chosen from 200 well-characterized clade C envelope (Env)-pseudotyped viruses from early infection. All 200 Envs were expressed as a single round of replication pseudoviruses and were tested to quantify neutralization titers by 16 broadly neutralizing antibodies (bnAbs) and sera from 30 subjects with chronic clade C infections. We selected large panels of 50 and 100 Envs either to characterize cross-reactive breadth for sera identified as having potent neutralization activity based on initial screening or to evaluate neutralization magnitude-breadth distributions of newly isolated antibodies. We identified these panels by downselection after hierarchical clustering of bnAb neutralization titers. The resulting panels represent the diversity of neutralization profiles throughout the range of virus sensitivities identified in the original panel of 200 viruses. A small 12-Env panel was chosen to screen sera from vaccine trials or natural-infection studies for neutralization responses. We considered panels selected by previously described methods but favored a computationally informed method that enabled selection of viruses representing diverse neutralization sensitivity patterns, given that we do not a priori know what the neutralization-response profile of vaccine sera will be relative to that of sera from infected individuals. The resulting 12-Env panel complements existing panels. Use of standardized panels enables direct comparisons of data from different trials and study sites testing HIV-1 clade C-specific products. HIV-1 group M includes nine clades and many recombinants. Clade C is the most common lineage, responsible for roughly half of current HIV-1 infections, and is a focus for vaccine design and testing. Standard reference reagents, particularly virus panels to study neutralization by antibodies, are crucial for developing cost-effective and yet rigorous and reproducible assays against diverse examples of this variable virus. We developed clade C-specific panels for use as standardized reagents to monitor complex polyclonal sera for neutralization activity and to characterize the potency and breadth of cross-reactive neutralization by monoclonal antibodies, whether engineered or isolated from infected individuals. We chose from 200 southern African, clade C envelope-pseudotyped viruses with neutralization titers against 16 broadly neutralizing antibodies and 30 sera from chronic clade C infections. We selected panels to represent the diversity of bnAb neutralization profiles and Env neutralization sensitivities. Finally, use of standard virus panels can facilitate comparison of results across studies and sites.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2013

Small Genomes and Sparse Metabolisms of Sediment-Associated Bacteria from Four Candidate Phyla

Cultivation-independent surveys of microbial diversity have revealed many bacterial phyla that lack cultured representatives. These lineages, referred to as candidate phyla, have been detected across many environments. Here, we deeply sequenced microbial communities from acetate-stimulated aquifer sediment to recover the complete and essentially complete genomes of single representatives of the candidate phyla SR1, WWE3, TM7, and OD1. All four of these genomes are very small, 0.7 to 1.2 Mbp, and have large inventories of novel proteins. Additionally, all lack identifiable biosynthetic pathways for several key metabolites. The SR1 genome uses the UGA codon to encode glycine, and the same codon is very rare in the OD1 genome, suggesting that the OD1 organism could also transition to alternate coding. Interestingly, the relative abundance of the members of SR1 increased with the appearance of sulfide in groundwater, a pattern mirrored by a member of the phylum Tenericutes . All four genomes encode type IV pili, which may be involved in interorganism interaction. On the basis of these results and other recently published research, metabolic dependence on other organisms may be widely distributed across multiple bacterial candidate phyla. Few or no genomic sequences exist for members of the numerous bacterial phyla lacking cultivated representatives, making it difficult to assess their roles in the environment. This paper presents three complete and one essentially complete genomes of members of four candidate phyla, documents consistently small genome size, and predicts metabolic capabilities on the basis of gene content. These metagenomic analyses expand our view of a lifestyle apparently common across these candidate phyla.

59 BASIC BIOLOGICAL SCIENCES↗
DOE OSTI2020

Deconstructing the Soil Microbiome into Reduced-Complexity Functional Modules

The soil microbiome is an invaluable component of the biosphere and critical for ecosystem functions, including biogeochemical cycling, soil-atmosphere gas exchange, degradation of toxic compounds, and promotion of plant growth and stress resistance/resilience. An improved understanding of the soil microbiome will help with predicting how these processes respond to external perturbations, and with harnessing beneficial aspects of the soil microbiome for agronomic applications such as crop amendments. However, the extensive taxonomic and functional diversity inherent within the soil microbiome hinders efficient analysis of this system. Microbial biodiversity in soils is orders of magnitude greater than other commonly studied systems such as the human gut microbiome (Blum, Zechmeister-Boltenstern, and Keiblinger 2019; Berendsen, Pieterse, and Bakker 2012). Thousands of microbial taxa may be found in a single gram of soil (Roesch et al. 2007), and high rates of gene flow and mutation further promote microbial diversification (Sergaki et al. 2018). Concomitantly, functional diversity in soil is similarly extensive. Soil is a heterogeneous mixture of microenvironments with defined physical and chemical attributes (Bach et al. 2018), within which numerous microbial guilds of distinct life-strategies and metabolic capacities can be found (Perez-Garcia, Lear, and Singhal 2016; H.-S. Song et al. 2014). Furthermore, the soil microbiome harbors a significant fraction of rare and/or quiescent members (Blagodatskaya and Kuzyakov 2013) that exist below the threshold of detection of current technologies. Such rare taxa may make significant contributions to process rates (Shade and Gilbert 2015; Dawson et al. 2017), but their scarcity complicates their identification and analysis. Finally, the vast amounts of information generated through holistic analyses of soil communities represent an intense computational burden (Scholz, Lo, and Chain 2012; Prosser 2015) that precludes assessment of the complete functional and taxonomic diversity contained in this ecosystem.

Naylor, Dan T.↗