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At least 235 records · Page 13

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DOE OSTI2014

Evaluation of Cloud-resolving and Limited Area Model Intercomparison Simulations using TWP-ICE Observations. Part 2: Rain Microphysics

Ten 3D cloud-resolving model (CRM) simulations and four 3D limited area model (LAM) simulations of an intense mesoscale convective system observed on January 23-24, 2006 during the Tropical Warm Pool – International Cloud Experiment (TWP-ICE) are compared with each other and with observations and retrievals from a scanning polarimetric radar, co-located UHF and VHF vertical profilers, and a Joss-Waldvogel disdrometer in an attempt to explain published results showing a low bias in simulated stratiform rainfall. Despite different forcing methodologies, similar precipitation microphysics errors appear in CRMs and LAMs with differences that depend on the details of the bulk microphysics scheme used. One-moment schemes produce too many small raindrops, which biases Doppler velocities low, but produces rain water contents (RWCs) that are similar to observed. Two-moment rain schemes with a gamma shape parameter (μ) of 0 produce excessive size sorting, which leads to larger Doppler velocities than those produced in one-moment schemes, but lower RWCs than observed. Two moment schemes also produce a convective median volume diameter distribution that is too broad relative to observations and thus, may have issues balancing raindrop formation, collision coalescence, and raindrop breakup. Assuming a μ of 2.5 rather than 0 for the raindrop size distribution improves one-moment scheme biases, and allowing μ to have values greater than 0 may improve two-moment schemes. Under-predicted stratiform rain rates are associated with under-predicted ice water contents at the melting level rather than excessive rain evaporation, in turn likely associated with convective detrainment that is too high in the troposphere and mesoscale circulations that are too weak. In addition to stronger convective updrafts than observed, limited domain size prevents a large, well-developed stratiform region from developing in CRMs, while a dry bias in ECMWF analyses does the same to the LAMs.

cloud-resolving↗

DOE OSTI2012

Characterization of cancer stem-like cells derived from a side population of a human gallbladder carcinoma cell line, SGC-996

Highlights: Black-Right-Pointing-Pointer We sorted SP cells from a human gallbladder carcinoma cell lines, SGC-996. Black-Right-Pointing-Pointer SP cells displayed higher proliferation and stronger clonal-generating capability. Black-Right-Pointing-Pointer SP cells showed more migratory and invasive abilities. Black-Right-Pointing-Pointer SP cells were more resistant and tumorigenic than non-SP counterparts. Black-Right-Pointing-Pointer ABCG2 might be a candidate as a marker for SP cells. -- Abstract: The cancer stem cell (CSC) hypothesis proposes that CSCs, which can renew themselves proliferate infinitely, and escape chemotherapy, become the root of recurrence and metastasis. Previous studies have verified that side population (SP) cells, characterized by their ability to efflux lipophilic substrate Hoechst 33342, to share many characteristics of CSCs in multiplying solid tumors. The purpose of this study was to sort SP cells from a human gallbladder carcinoma cell line, SGC-996 and to preliminarily identify the biological characteristics of SP cells from the cell line. Using flow cytometry we effectively sorted SP cells from the cell line SGC-996. SP cells not only displayed higher proliferative, stronger clonal-generating, more migratory and more invasive capacities, but showed stronger resistance. Furthermore, our experiments demonstrated that SP cells were more tumorigenic than non-SP counterparts in vivo. Real-time PCR analysis and immunocytochemistry showed that the expression of ATP-binding cassette subfamily G member 2 (ABCG2) was significantly higher in SP cells. Hence, these results collectively suggest that SP cells are progenitor/stem-like cells and ABCG2 might be a candidate marker for SP cells in human gallbladder cancer.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2014

Hepatocyte growth factor regulated tyrosine kinase substrate in the peripheral development and function of B-cells

Highlights: •ESCRT-0 protein regulates the development of peripheral B-cells. •BCR expression on cell surface should be controlled by the endosomal-sorting system. •Hrs plays important roles in responsiveness to Ag stimulation in B lymphocytes. -- Abstract: Hepatocyte growth factor (HGF)-regulated tyrosine kinase substrate (Hrs) is a vesicular sorting protein that functions as one of the endosomal-sorting proteins required for transport (ESCRT). Hrs, which binds to ubiquitinated proteins through its ubiquitin-interacting motif (UIM), contributes to the lysosomal transport and degradation of ubiquitinated membrane proteins. However, little is known about the relationship between B-cell functions and ESCRT proteins in vivo. Here we examined the immunological roles of Hrs in B-cell development and functions using B-cell-specific Hrs-deficient (Hrs{sup flox/flox};mb1{sup cre/+}:Hrs-cKO) mice, which were generated using a cre-LoxP recombination system. Hrs deficiency in B-cells significantly reduced T-cell-dependent antibody production in vivo and impaired the proliferation of B-cells treated in vitro with an anti-IgM monoclonal antibody but not with LPS. Although early development of B-cells in the bone marrow was normal in Hrs-cKO mice, there was a significant decrease in the number of the peripheral transitional B-cells and marginal zone B-cells in the spleen of Hrs-cKO mice. These results indicate that Hrs plays important roles during peripheral development and physiological functions of B lymphocytes.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2018

The galectin LEC-5 is a novel binding partner for RAB-11

RAB-11/Rab11 is an endosomal GTPase with conserved roles in directional trafficking and apical domain formation in polarized epithelial cells. From a yeast two-hybrid screen using full-length C. elegans RAB-11 as bait, we identified LEC-5 as a novel binding protein for RAB-11. LEC-5 is an ortholog of mammalian Galectin-9 which associates with glycosphingolipids and is implicated in apical cargo sorting. We further confirmed the interaction between RAB-11 and LEC-5 via GST-pull down, co-immunoprecipitation and bimolecular fluorescence complementation. In addition, we showed that LEC-5 binds to RAB-11 with its C-terminus. Our results indicate a novel role of RAB-11 in apical sorting via LEC-5. Such a role would extend RAB-11's function as a master regulator of apical trafficking and suggest it could translate apical sorting signals into apical vesicle directionality.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2011

HRTEM and neutron diffraction study of Li{sub x}Mo{sub 5}O{sub 17}: From the ribbon (x=5) structure to the rock salt (x=12) structure

Structure determination of the fully intercalated phase Li{sub 12}Mo{sub 5}O{sub 17} and of the deintercalated oxide Li{sub 5}Mo{sub 5}O{sub 17} has been carried out by electron microscopy and neutron powder diffraction. The reversible topotactic transformation between the ordered rock salt structure of the former and the ribbon structure of the latter (closely related to that of Li{sub 4}Mo{sub 5}O{sub 17}) is explained on the following basis: both structures can be described as strips built up as an assembly of infinite ribbons of MoO{sub 6} octahedra that are five octahedra thick, and that differ by slight displacements of the octahedral ribbons. We show that the electrochemical behavior of the Li{sub x}Mo{sub 5}O{sub 17} system is based on two sorts of Li{sup +} sites; those that are located within the strips between the ribbons, and those that are located at the border of the strips. The high rate of Li intercalation in this oxide and its reversibility are discussed in terms of its peculiar structure. -- Graphical abstract: Structure determination of the fully intercalated phase Li{sub 12}Mo{sub 5}O{sub 17} and of the deintercalated oxide Li{sub 5}Mo{sub 5}O{sub 17} has been carried out by electron microscopy and neutron powder diffraction. The reversible topotactic transformation between the ordered rock salt structure of the former and the ribbon structure of the latter is explained on the following basis: both structures can be described as strips built up as an assembly of infinite ribbons of MoO{sub 6} octahedra that are five octahedra thick, and that differ by slight displacements of the octahedral ribbons. We show that the electrochemical behavior of the Li{sub x}Mo{sub 5}O{sub 17} system is based on two sorts of Li{sup +} sites; those that are located within the strips between the ribbons, and those that are located at the border of the strips. The high rate of Li intercalation in this oxide and its reversibility are discussed in terms of its peculiar structure. Research highlights: {yields} Electron microscopy and neutron powder diffraction structure determination {yields} We have explained the reversible topotactic transformation between an ordered rock salt structure and a ribbon structure {yields} We show that the electrochemical behavior of the Li{sub x}Mo{sub 5}O{sub 17} system is based on two sorts of Li{sup +} sites {yields} The high rate of Li intercalation in this oxide and its reversibility are discussed in terms of its peculiar structure.

36 MATERIALS SCIENCE↗

DOE OSTI2015

Separation of colloidal two dimensional materials by density gradient ultracentrifugation

Two-dimensional (2D) materials have been made through various approaches but obtaining monodispersed simply by synthesis optimization gained little success, which highlighted the need for introducing nanoseparation methods. Density gradient ultracentrifugation method has emerged as a versatile and scalable method for sorting colloidal 2D nanomaterials. Isopycnic separation was applied on thickness-dependent separation of graphene nanosheets. And rate-zonal separation, as a more versatile separation method, demonstrated its capability in sorting nanosheets of chemically modified single layered graphene, layered double hydroxide, and even metallic Ag. Establishing such density gradient ultracentrifugation method not only achieves monodispersed nanosheets and provides new opportunities for investigation on size dependent properties of 2D materials, but also makes the surface modification possible by introducing “reaction zones” during sedimentation of the colloids. - Graphical abstract: Two-dimensional (2D) materials have been made through various approaches but obtaining monodispersed simply by synthesis optimization gained little success, which highlighted the need for introducing nanoseparation methods. Density gradient ultracentrifugation method has emerged as a versatile and scalable method for sorting colloidal 2D nanomaterials according to their size of thickness difference. Establishing such density gradient ultracentrifugation method not only achieves monodispersed nanosheets and provides new opportunities for investigation on size dependent properties of 2D materials, but also makes the surface modification possible by introducing “reaction zones” during sedimentation of the colloids. - Highlights: • Density gradient ultracentrifugation was applied on size separation of 2D material. • Isopycnic separation was applied on separation of low density materials. • Rate-zonal separation was applied on separation of large density materials. • Size separation lead to investigation on size dependent properties of 2D material. • Mechanism was investigated by introducing “reaction zones” in the gradient.

77 NANOSCIENCE AND NANOTECHNOLOGY↗

DOE OSTI2015

Improvement of the management of residual waste in areas without thermal treatment facilities: A life cycle analysis of an Italian management district

Highlights: • LCA analysis of two option for residual waste management. • Exploitation of mechanical physical sorting facility for extracting recyclable from RMSW. • Processing the mechanically sorted organic fraction in bioreactor landfill. • Sensitivity analysis demonstrate high influence for impact assessment of substitution ratio for recycle materials. - Abstract: Starting from an existing waste management district without thermal treatment facilities, two different management scenarios for residual waste were compared by life cycle assessment (LCA). The adoption of a bioreactor landfill for managing the mechanically sorted organic fraction instead of bio-stabilization led to reduction of global warming and fresh water eutrophication by 50% and 10%, respectively. Extraction of recyclables from residual waste led to avoided emissions for particulate matter, acidification and resource depletion impact categories. Marginal energy and the amount of energy recovered from landfill gas marginally affected the LCA results. On the contrary the quality of the recyclables extracted can significantly modify the eco profile of the management schemes.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2017

Environmental performances of different configurations of a material recovery facility in a life cycle perspective

Highlights: • An attributional LCA assessed the environmental performances of an integrated MRF. • Different configurations of the MRF technological evolution have been analysed. • Reliable data provide a large LCI table for all the configurations and each sub-units. • A biological treatment in-situ improves the potentials of land occupation and global warming. • The inclusion of a small-scale gasifier could improve almost all the environmental potentials. - Abstract: The study evaluated the environmental performances of an integrated material recovery facility (MRF) able to treat 32 kt/y of unsorted mixed waste, made of residuals from household source separation and separate collection. The facility includes a mechanical sorting platform for the production of a solid recovered fuel (SRF) utilized in an external waste-to-energy plant, bio-cells for tunnel composting of organic fraction, and a sanitary landfill for the safe disposal of ultimate waste. All the MRF sub-units have been analysed in depth in order to acquire reliable data for a life cycle assessment study, focused on the environmental performances of different configurations of the facility. The study investigated a “past” configuration, including just mechanical sorting, landfilling and biogas combustion in a gas engine, and the “present” one, which includes also a composting unit. Two possible “future” configurations, having a gasifier inside the MRF battery limits, have been also analysed, assessing the performances of two fluidized bed reactors of different size, able to gasify only the residues generated by the sorting platform or the whole amount of produced SRF, respectively. The analysis evaluated the contributions of each unit in the different configurations and allowed a reliable assessment of the technological evolution of the facility. The results quantified the positive effect of the inclusion of an aerobic treatment of the waste organic fraction. The SRF gasification in situ appears to improve the MRF environmental performances in all the impact categories, with the exclusion of that of global warming.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

DOE OSTI2024

Assessment of Different Application Grades of Post-Consumer Recycled (PCR) Polyolefins from Material Recovery Facilities (MRFs) in the United States

The recycling of plastics has been gaining traction in the following years as many companies, organizations, and governments are pushing toward a more circular economy. However, plastic recycling rates are still small compared to those of plastic production, especially for polyolefins, such as high-density polyethylene (HDPE) and polypropylene (PP). With the goal of finding applications for recycled plastic resin, recyclers can face some challenges associated with the heterogeneity in the polymer properties of plastic waste streams, which could hinder the full potential of recycling due to the lack of a consistent feedstock. Here, this study assessed how an additional sorting step of recycled HDPE and PP streams, separating them into two categories based on the previous application, impacts the recycled polymer properties, such as physical, molecular, and thermal. It was demonstrated that manually separating HDPE articles previously processed by extrusion blow molding (EBM) or injection molding (IM) and PP articles by thermoforming (TF) or IM could yield recycled resins with more consistent properties, considering the significant differences between them. This highlights that an additional sorting step would allow recyclers to target more premium or currently unattainable plastic processing applications with postconsumer recycled (PCR) resins, which could significantly promote an increase in plastic recycling rates. Nevertheless, the development of new technologies is needed to automate this extra sorting step, as with the increase in scale manual sortation could not be feasible and cost-effective, reducing the attractiveness of using PCR resins in comparison to virgin ones.

36 MATERIALS SCIENCE↗

DOE OSTI2019

Function-driven single-cell genomics uncovers cellulose-degrading bacteria from the rare biosphere

Assigning a functional role to a microorganism has historically relied on cultivation of isolates or detection of environmental genome-based biomarkers using a posteriori knowledge of function. However, the emerging field of function-driven single-cell genomics aims to expand this paradigm by identifying and capturing individual microbes based on their in situ functions or traits. To identify and characterize yet uncultivated microbial taxa involved in cellulose degradation, we developed and benchmarked a function-driven single-cell screen, which we applied to a microbial community inhabiting the Great Boiling Spring (GBS) Geothermal Field, northwest Nevada. Our approach involved recruiting microbes to fluorescently labeled cellulose particles, and then isolating single microbe-bound particles via fluorescence-activated cell sorting. The microbial community profiles prior to sorting were determined via bulk sample 16S rRNA gene amplicon sequencing. The flow-sorted cellulose-bound microbes were subjected to whole genome amplification and shotgun sequencing, followed by phylogenetic placement. Next, putative cellulase genes were identified, expressed and tested for activity against derivatives of cellulose and xylose. Alongside typical cellulose degraders, including members of the Actinobacteria, Bacteroidetes, and Chloroflexi, we found divergent cellulases encoded in the genome of a recently described candidate phylum from the rare biosphere, Goldbacteria, and validated their cellulase activity. As this genome represents a species-level organism with novel and phylogenetically distinct cellulolytic activity, we propose the name Candidatus ‘Cellulosimonas argentiregionis’. We expect that this function-driven single-cell approach can be extended to a broad range of substrates, linking microbial taxonomy directly to in situ function.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2023

Effects of grain size and small-scale bedform architecture on CO 2 saturation from buoyancy-driven flow

Small-scale (mm-dm scale) heterogeneity has been shown to significantly impact CO 2 migration and trapping. To investigate how and why different aspects of small-scale heterogeneity affect the amount of capillary trapping during buoyancy-driven upward migration of CO 2 , we conducted modified invasion percolation simulations on heterogeneous domains. Realistic simulation domains are constructed by varying two important aspects of small-scale geologic heterogeneity: sedimentary bedform architecture and grain size contrast between the matrix and the laminae facies. Buoyancy-driven flow simulation runs cover 59 bedform architecture and 40 grain size contrast cases. Simulation results show that the domain effective CO 2 saturation is strongly affected by both grain size and bedform architecture. At high grain size contrasts, bedforms with continuous ripple lamination at the cm scale tend to retain higher CO 2 saturation than bedforms with discontinuous or cross lamination. In addition, the “extremely well sorted” grain sorting cases tend to have lower CO 2 saturation than expected for cross-laminated domains. Finally, both a denser CO 2 phase and greater interfacial tension increase CO 2 saturation. Again, variation in fluid properties seems to have a greater effect on CO 2 saturation for cross-laminated domains. This result suggests that differences in bedform architecture can impact how CO 2 saturation values respond to other variables such as grain sorting and fluid properties.

58 GEOSCIENCES↗

DOE OSTI2024

The Pixel Anomaly Detection Tool : a user-friendly GUI for classifying detector frames using machine-learning approaches

Data collection at X-ray free electron lasers has particular experimental challenges, such as continuous sample delivery or the use of novel ultrafast high-dynamic-range gain-switching X-ray detectors. This can result in a multitude of data artefacts, which can be detrimental to accurately determining structure-factor amplitudes for serial crystallography or single-particle imaging experiments. Here, a new data-classification tool is reported that offers a variety of machine-learning algorithms to sort data trained either on manual data sorting by the user or by profile fitting the intensity distribution on the detector based on the experiment. This is integrated into an easy-to-use graphical user interface, specifically designed to support the detectors, file formats and software available at most X-ray free electron laser facilities. The highly modular design makes the tool easily expandable to comply with other X-ray sources and detectors, and the supervised learning approach enables even the novice user to sort data containing unwanted artefacts or perform routine data-analysis tasks such as hit finding during an experiment, without needing to write code.

47 OTHER INSTRUMENTATION↗

DOE OSTI2013

Respiratory triggered 4D cone-beam computed tomography: A novel method to reduce imaging dose

Purpose: A novel method called respiratory triggered 4D cone-beam computed tomography (RT 4D CBCT) is described whereby imaging dose can be reduced without degrading image quality. RT 4D CBCT utilizes a respiratory signal to trigger projections such that only a single projection is assigned to a given respiratory bin for each breathing cycle. In contrast, commercial 4D CBCT does not actively use the respiratory signal to minimize image dose. Methods: To compare RT 4D CBCT with conventional 4D CBCT, 3600 CBCT projections of a thorax phantom were gathered and reconstructed to generate a ground truth CBCT dataset. Simulation pairs of conventional 4D CBCT acquisitions and RT 4D CBCT acquisitions were developed assuming a sinusoidal respiratory signal which governs the selection of projections from the pool of 3600 original projections. The RT 4D CBCT acquisition triggers a single projection when the respiratory signal enters a desired acquisition bin; the conventional acquisition does not use a respiratory trigger and projections are acquired at a constant frequency. Acquisition parameters studied were breathing period, acquisition time, and imager frequency. The performance of RT 4D CBCT using phase based and displacement based sorting was also studied. Image quality was quantified by calculating difference images of the test dataset from the ground truth dataset. Imaging dose was calculated by counting projections. Results: Using phase based sorting RT 4D CBCT results in 47% less imaging dose on average compared to conventional 4D CBCT. Image quality differences were less than 4% at worst. Using displacement based sorting RT 4D CBCT results in 57% less imaging dose on average, than conventional 4D CBCT methods; however, image quality was 26% worse with RT 4D CBCT. Conclusions: Simulation studies have shown that RT 4D CBCT reduces imaging dose while maintaining comparable image quality for phase based 4D CBCT; image quality is degraded for displacement based RT 4D CBCT in its current implementation.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2014

SU-D-17A-07: Development and Evaluation of a Prototype Ultrasonography Respiratory Monitoring System for 4DCT Reconstruction

Purpose: Respiratory motion artifacts are commonly seen in the abdominal and thoracic CT images. A Real-time Position Management (RPM) system is integrated with CT simulator using abdominal surface as a surrogate for tracking the patient respiratory motion. The respiratory-correlated four-dimensional computed tomography (4DCT) is then reconstructed by GE advantage software. However, there are still artifacts due to inaccurate respiratory motion detecting and sorting methods. We developed an Ultrasonography Respiration Monitoring (URM) system which can directly monitor diaphragm motion to detect respiratory cycles. We also developed a new 4DCT sorting and motion estimation method to reduce the respiratory motion artifacts. The new 4DCT system was compared with RPM and the GE 4DCT system. Methods: Imaging from a GE CT scanner was simultaneously correlated with both the RPM and URM to detect respiratory motion. A radiation detector, Blackcat GM-10, recorded the X-ray on/off and synchronized with URM. The diaphragm images were acquired with Ultrasonix RP system. The respiratory wave was derived from diaphragm images and synchronized with CT scanner. A more precise peaks and valleys detection tool was developed and compared with RPM. The motion is estimated for the slices which are not in the predefined respiratory phases by using block matching and optical flow method. The CT slices were then sorted into different phases and reconstructed, compared with the images reconstructed from GE Advantage software using respiratory wave produced from RPM system. Results: The 4DCT images were reconstructed for eight patients. The discontinuity at the diaphragm level due to an inaccurate identification of phases by the RPM was significantly improved by URM system. Conclusion: Our URM 4DCT system was evaluated and compared with RPM and GE 4DCT system. The new system is user friendly and able to reduce motion artifacts. It also has the potential to monitor organ motion during therapy.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

DOE OSTI2014

TH-E-17A-07: Improved Cine Four-Dimensional Computed Tomography (4D CT) Acquisition and Processing Method

Purpose: Artifacts arising from the 4D CT acquisition and post-processing methods add systematic uncertainty to the treatment planning process. We propose an alternate cine 4D CT acquisition and post-processing method to consistently reduce artifacts, and explore patient parameters indicative of image quality. Methods: In an IRB-approved protocol, 18 patients with primary thoracic malignancies received a standard cine 4D CT acquisition followed by an oversampling 4D CT that doubled the number of images acquired. A second cohort of 10 patients received the clinical 4D CT plus 3 oversampling scans for intra-fraction reproducibility. The clinical acquisitions were processed by the standard phase sorting method. The oversampling acquisitions were processed using Dijkstras algorithm to optimize an artifact metric over available image data. Image quality was evaluated with a one-way mixed ANOVA model using a correlation-based artifact metric calculated from the final 4D CT image sets. Spearman correlations and a linear mixed model tested the association between breathing parameters, patient characteristics, and image quality. Results: The oversampling 4D CT scans reduced artifact presence significantly by 27% and 28%, for the first cohort and second cohort respectively. From cohort 2, the inter-replicate deviation for the oversampling method was within approximately 13% of the cross scan average at the 0.05 significance level. Artifact presence for both clinical and oversampling methods was significantly correlated with breathing period (ρ=0.407, p-value<0.032 clinical, ρ=0.296, p-value<0.041 oversampling). Artifact presence in the oversampling method was significantly correlated with amount of data acquired, (ρ=-0.335, p-value<0.02) indicating decreased artifact presence with increased breathing cycles per scan location. Conclusion: The 4D CT oversampling acquisition with optimized sorting reduced artifact presence significantly and reproducibly compared to the phase-sorted clinical acquisition.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2015

SU-F-303-02: Achieving 4D MRI in Regular Breathing Cycle with Extended Acquisition Time of Dynamic MR Images

Purpose: The purpose of this study is to develop a technique to obtain four-dimensional (4D) magnetic resonance (MR) images that are more representative of a patient’s typical breathing cycle by utilizing an extended acquisition time while minimizing the image artifacts. Methods: The 4D MR data were acquired with the balanced steady state free precession in two-dimensional sagittal plane of view. Each slice was acquired repeatedly for about 15 s, thereby obtaining multiple images at each of the 10 phases in the respiratory cycle. This improves the probability that at least one of the images were acquired at the desired phase during a regular breathing cycle. To create optimal 4D MR images, an iterative approach was used to identify the set of images that yielded the highest slice-to-slice similarity. To assess the effectiveness of the approach, the data set was truncated into periods of 7 s (50 time points), 11 s (75 time points) and the full 15 s (100 time points). The 4D MR images were then sorted with data of the three different acquisition periods for comparison. Results: In general, the 4D MR images sorted using data from longer acquisition periods showed less mismatched artifacts. In addition, the normalized cross correlation (NCC) between slices of a 4D volume increases with increased acquisition period. The average NCC was 0.791 from the 7 s period, 0.794 from the 11 s period and 0.796 from the 15 s period. Conclusion: Our preliminary study showed that extending the acquisition time with the proposed sorting technique can improve image quality and reduce artifact presence in the 4D MR images. Data acquisition over two breathing cycles is a good trade-off between artifact reduction and scan time. This research was partially funded by the the Center for Radiation Oncology Research from UT MD Anderson Cancer Center.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2019

A Strongly Fluorescing Anaerobic Reporter and Protein-Tagging System for Clostridium Organisms Based on the Fluorescence-Activating and Absorption-Shifting Tag Protein (FAST)

Visualizing protein localization and characterizing gene expression activity in live Clostridium cells is limited for lack of a real-time, highly fluorescent, oxygen-independent reporter system. Enzymatic reporter systems have been used successfully for many years with Clostridium spp.; however, these assays do not allow for real-time analysis of gene expression activity with flow cytometry or for visualizing protein localization through fusion proteins. Commonly used fluorescent reporter proteins require oxygen for chromophore maturation and cannot be used for most strictly anaerobic Clostridium organisms. Here we show that the fluorescence-activating and absorption-shifting tag protein (FAST), when associated with the fluorogenic ligand 4-hydroxy-3-methylbenzylidene-rhodanine (HMBR; now commercially available) and other commercially available ligands, is highly fluorescent in Clostridium acetobutylicum under anaerobic conditions. Using flow cytometry and a fluorescence microplate reader, we demonstrated FAST as a reporter system by employing the promoters of the C. acetobutylicum thiolase (thl), acetoacetate decarboxylase (adc), and phosphotransbutyrylase (ptb) metabolic genes, as well as a mutant P thl and modified ribosome binding site (RBS) versions of P adc and P ptb . Flow cytometry-based sorting was efficient and fast in sorting FAST-expressing cells, and positively and negatively sorted cells could be effectively recultured. FAST was also used to tag and examine protein localization of the predicted cell division FtsZ partner protein, ZapA, to visualize the divisome localization in live C. acetobutylicum cells. Our findings suggest that FAST can be used to further investigate Clostridium divisomes and more broadly the localization and expression levels of other proteins in Clostridium organisms, thus enabling cell biology studies with these organisms.

59 BASIC BIOLOGICAL SCIENCES↗

DOE OSTI2011

Image Composition Engine for Tiles

The Image Composition Engine for Tiles (lceT) is a high-performance sort-last parallel rendering library. It is designed to be used in parallel applications requiring rendering. The primary purpose of IceT is to be integrated into parallel visualization applications such as ParaView to provide parallel rendering capabilities. The Image Composition Engine for Tiles (lceT) is a high-performance sort-last parallel rendering library. IceT uses a "sort-Iasf' approach to rendering. Each process in a parallel application independently renders a local piece of geometry. The resulting images are given to IceT, and IceT combines the images together to form a single cohesive image. Ice T is also capable of driving tiled displays, largeformat displays comprising an array of smaller displays. To this end IceT can collect the smaller tile images and organize them such that the entire tiled display can be driven. Ice T takes advantage of spatial coherence in geometry by identifying empty regions of the display and reducing the overall required work.

Moreland, Kenneth↗