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NASA NTRS2017

A Programmable SDN+NFV Architecture for UAV Telemetry Monitoring

With the explosive growth in UAV numbers forecast worldwide, a core concern is how to manage the ad-hoc network configuration required for mobility management. As UAVs migrate among ground control stations, associated network services, routing and operational control must also rapidly migrate to ensure a seamless transition. In this paper, we present a novel, lightweight and modular architecture which supports high mobility, resilience and flexibility through the application of SDN and NFV principles on top of the UAV infrastructure. By combining SDN programmability and Network Function Virtualization we can achieve resilient infrastructure migration of network services, such as network monitoring and anomaly detection, coupled with migrating UAVs to enable high mobility management. Our container-based monitoring and anomaly detection Network Functions (NFs) can be tuned to specific UAV models providing operators better insight during live, high-mobility deployments. We evaluate our architecture against telemetry from over 80flights from a scientific research UAV infrastructure.

Resilience↗

NASA NTRSDate not supplied

Influence of Microstructural Features on Austenite-Martensite Interfaces in NiTi Shape Memory Alloys

Shape memory alloys (SMAs) exhibit several unique thermomechanical properties due to a reversible martensitic phase transformation. A key aspect of this transformation is the austenite-martensite interface. Here we present a molecular dynamics (MD) simulation methodology to study the atomic-scale features of austenite-martensite interface migration under near-equilibrium conditions. In single crystals, the interfaces migrate rapidly with only a small thermodynamic driving force. In polycrystals, however, interface migration is significantly impeded due to the change in orientation relationship at grain boundaries and the stored elastic energy resulting from microstructural constraints. This behavior can be linked to several mechanisms associated with transformation width and hysteresis in SMAs, properties of great importance for applications involving actuation. Additionally, we will present preliminary MD simulation results on the influence of precipitates on the formation and migration of austenite-martensite interfaces.

Gabriel Plummer↗

NASA NTRS2024

Molecular Dynamics Simulations of Microstructural Effects on Austenite-Martensite Interfaces in NiTi

Formation and migration of austenite-martensite interfaces plays the key role in reversible martensitic transformations of shape memory alloys (SMAs). How these interfaces interact with the SMA microstructure is a primary determining factor in important functional properties such as hysteresis and transformation span. As such, successful microstructural engineering of SMAs requires in-depth knowledge of interface behavior. The rapid nature of martensitic transformations makes experimental observations of moving austenite-martensite interfaces challenging. Molecular dynamics (MD) simulation is a unique tool which can probe the atomic-scale details of austenite-martensite interfaces as they migrate through different microstructures. However, in focusing on the entire transformation process, including the nucleation of new phases, MD studies are usually performed so far from equilibrium that their relevance to experiment is questionable. Here, we demonstrate new MD techniques to generate energetically preferred austenite-martensite interfaces in NiTi under near-equilibrium conditions. The interfaces are semi-coherent, exhibiting a series of structural disconnections, and they can migrate rapidly through single crystals under only small thermodynamic driving forces. In contrast, when interfaces migrate in polycrystals, their motion is impeded by thermoelastic effects as well as changes in orientation relationships at grain boundaries. Microstructures which accumulate large amounts of elastic energy tend to release some fraction through irreversible, hysteresis-inducing mechanisms. We demonstrate that engineering microstructures with less constraints is a viable strategy to produce SMAs with reduced hysteresis and transformation span. Similar thermoelastic and hysteresis-inducing mechanisms also arise when austenite-martensite interfaces encounter precipitates and can be controlled by tuning characteristics of the precipitate distribution.

Gabriel Plummer↗

NASA NTRSDate not supplied

Molecular Dynamics Simulations of Microstructural Effects on Austenite-Martensite Interfaces in NiTi

The formation and migration of austenite-martensite interfaces plays the key role in reversible martensitic transformations of shape memory alloys (SMAs). How these interfaces interact with the SMA microstructure is a primary determining factor in important functional properties such as hysteresis and transformation span. As such, successful microstructural engineering of SMAs requires in-depth knowledge of interface behavior. The rapid nature of martensitic transformations makes experimental observations of moving austenite-martensite interfaces challenging. Molecular dynamics (MD) simulation is a unique tool which can probe the atomic-scale details of austenite-martensite interfaces as they migrate through different microstructures. However, in focusing on the entire transformation process, including the nucleation of new phases, MD studies are usually performed so far from equilibrium that their relevance to experiment is questionable. Here, we demonstrate new MD techniques to generate energetically preferred austenite-martensite interfaces in NiTi under near-equilibrium conditions. The interfaces are semi-coherent, exhibiting a series of structural disconnections, and they can migrate rapidly through single crystals under only small thermodynamic driving forces. In contrast, when interfaces migrate in polycrystals, their motion is impeded by thermoelastic effects as well as changes in orientation relationships at grain boundaries. Microstructures which accumulate large amounts of elastic energy tend to release some fraction through irreversible, hysteresis-inducing mechanisms. We demonstrate that engineering microstructures with less constraints is a viable strategy to produce SMAs with reduced hysteresis and transformation span. Similar thermoelastic and hysteresis-inducing mechanisms also arise when austenite-martensite interfaces encounter precipitates and can be controlled by tuning characteristics of the precipitate distribution.

Gabriel Plummer↗

NASA NTRSDate not supplied

Influence of Microstructural Features on Austenite-Martensite Interfaces in NiTi Shape Memory Alloys

Gabriel Plummer↗

NASA NTRSDate not supplied

Molecular Dynamics Simulations of Microstructural Effects on Austenite-Martensite Interfaces in NiTi

Gabriel Plummer↗

DOE OSTI2019

Subtask 1.1 – Advanced Characterization of Unconventional Oil and Gas Reservoirs to Enhance CO2 Storage Resource Estimates

The Energy & Environmental Research Center (EERC) developed methods and approaches for advanced characterization of unconventional oil and gas reservoirs to enhance CO2 storage resource estimates using samples collected from organic-rich shales of the Bakken Formation. The project was funded under U.S. Department of Energy (DOE) Cooperative Agreement No. DE-FE0024233, with nonfederal cost share provided by Hitachi High Technologies America. The project participants included the EERC, Hitachi High Technologies America, the team at the National Energy Technology Laboratory’s (NETL’s) computed tomography (CT) scanning laboratory in Morgantown, West Virginia, and several NETL staff who have worked on CO2 storage resource estimation. Investigations on the use of subsurface geologic formations for CO2 storage have been ongoing for many years. In the last decade, improvements in directional drilling technology and hydraulic fracturing of organic-rich shale and other tight rock formations have opened new opportunities for CO2 storage, with the potential cobenefit of enhancing gas and oil recovery. Gas-bearing, organic-rich shales have been of particular interest as a carbon storage target because their ability to store large volumes of CO2 is implied by their ability to store methane; however, estimating the CO2 storage resource of organic-rich shales and other unconventional reservoirs is challenging because of the lack of detailed geologic and petrophysical data that are needed to develop improved volumetric equations. According to NETL’s method for estimating regional-scale CO2 storage resources, the key factors that control CO2 storage in geologic formations include void space within induced fractures, natural fractures and matrix pores, as well as the occurrence and distribution of organic matter (OM) and clay minerals. A key goal of this effort was to use field emission scanning electron microscopy (FESEM) analysis of fine-grain unconventional reservoir rocks coupled with advanced image analysis and machine learning techniques to quantify the occurrence and distribution of porosity, OM, and clay minerals. By better understanding the potential migration pathways of CO2 within tight rocks and quantifying the occurrence of sorptive components (OM and clays) along potential flow pathways, methods to estimate the CO2 storage resource of such rocks can be refined. The Bakken petroleum system (Bakken) was used as the source of rock samples for this effort because it includes organic-rich shales with contrasting degrees of thermal maturity as well as tight, nonshale clastic rocks with varying clay mineral content. To better understand the key components of interest in tight rock samples, the EERC worked closely with Hitachi to optimize the FESEM settings necessary to resolve the sample mineralogy, porosity and OM content at micro- to nanoscale resolution. The EERC also worked closely with Hitachi to apply and improve its advanced mineral identification and characterization system (AMICS) software so that it is better suited for mineral identification at the high resolution necessary to resolve features of interest in fine-grained rocks such as organic-rich shales. The EERC also applied advanced image analysis software packages, including Ilastik, to classify and quantify the key components of interest within the samples using FESEM imagery. These advanced techniques allowed for quantification and differentiation of the void space within fractures, pores and OM, as well as a technique to determine and quantify the key mineral and OM components surrounding pore spaces. Being able to better quantify the void space and the potential accessibility of CO2 to sorptive minerals or OM within tight rocks allows for improved estimation of the CO2 storage resource potential of organic-rich shales and other unconventional formations. The information derived from the image analysis techniques was used to refine NETL’s method for estimating regional-scale CO2 storage resources in collaboration with staff from NETL. An additional goal of this work was to better understand rates of CO2 permeation, flow pathways, and sorption in organic-rich shales and nonshale reservoir rocks and to evaluate potential changes in CO2 flow over time. The EERC conducted long-duration (several weeks) CO2 flow-through testing at reservoir conditions using Bakken shale and nonshale samples to assess CO2 permeation rates and retention/sorption within core plugs to represent bulk matrix behavior. Conventional isothermal adsorption testing over a range of pressure conditions was also performed on pulverized Bakken samples to better assess potential maximum sorptive capacity of the samples. The results of this testing illustrated the higher sorptive capacity of the Bakken shales when compared to the sorptive capacity of the nonshale reservoir rock, which contains very little OM when compared to the Bakken shales. The results also illustrate the differences in permeation behavior as a function of fracture flow versus matrix flow. In an attempt to visualize CO2 permeation and flow behavior in Bakken shale samples over time, the EERC collaborated with NETL’s CT Lab in Morgantown, West Virginia. A variety of tests were performed at the NETL lab on unfractured and fractured samples of the Bakken shale. The unfractured Bakken shale samples did not result in CO2 migration within the samples over the testing duration; thus imagery of CO2 migration pathways within the matrix of Bakken samples was not obtained. The results of the flow-through testing within fractured samples illustrate changes in CO2 permeation rates over time, with a trend of decreasing CO2 flow over time. The results of this effort suggest that FESEM analysis coupled with advanced image analysis and machine learning techniques are effective in characterizing and quantifying the mineralogy, porosity, fractures, and OM content of 2-D imagery at the high resolution needed to identify features of interest in tight rocks. In addition, the image analysis techniques allow for identification and quantification of different porosity types as well as the key mineral components and/or OM that CO2 would contact in the reservoir as it migrates through pores and fracture networks. This is key information needed to better estimate the CO2 storage resource potential of organic-rich shales and other tight reservoirs. Additional work is needed to determine the sorptive capacity of the individual sample components (OM and clay minerals) at reservoir conditions so that this approach and the refined CO2 storage resource equation can be applied to other shale and unconventional reservoirs. In addition, subsequent work should be performed to better understand the number of images needed to use stereological approaches to estimate the volume of sample components using 2-D imagery, to explore machine learning techniques to differentiate between induced vs. natural fractures, and to estimate the connectivity of pores and fracture networks using 2-D imagery. Being able to derive this information at the microscale would facilitate scaling up of the sample characterization data used for reservoir modeling and simulation to estimate the CO2 storage resource potential of tight formations.

03 NATURAL GAS↗

DOE OSTI2019

Postirradiation Examination of the ATF-1 Experiments - 2019 Status

A collaborative effort of the Advanced Fuels Campaign together with industry consortia is focused on the development of enhanced accident tolerant fuels for Light Water Reactors (LWRs). The scoping studies, referred to as ATF-1 irradiations, are being performed in the Advanced Test Reactor of Idaho National Laboratory using drop-in style irradiations. The Post-Irradiation Examinations (PIEs) of ATF-1 began in 2017 and continued in 2019. This report describes the results of examinations on ATF-1 irradiations designed to test the performance of advanced iron-based cladding, including compatibility of FeCrAl with UO2, and ATF-1 irradiation of U3Si5. Two rodlets with commercial iron-based cladding and UO2 pellets have been irradiated to low burnup (i.e., < 20 GWd/tHM). The first rodlet had an Alloy33 cladding, while the other had a Kanthal APMT® cladding. The performed PIEs were focused on the determination of fuel microstructure, evaluation of fuel-cladding interaction and irradiation-induced variations of cladding mechanical properties. For both rodlets, no significant changes in the cladding hardness were measured, and the cladding hoop strain remained limited. Gamma tomography revealed no Cs axial migration as expected for LWR fuel, but significant cesium radial migration, which was induced by thermal migration consequent to high irradiation temperatures. Microstructure analyses revealed formation of secondary phases along the pellet rim of samples from both rodlets. No wastage or extensive chemical interaction between the fuel and the internal wall of the cladding was observed, suggesting that the secondary phases accumulated at the pellet rim might be due to formation of Cs-U-O compounds. However, further advanced PIEs are necessary to finally confirm this hypothesis. Optical microscopy analysis of the rodlet designed by Oak Ridge National Laboratory to evaluate the interaction between UO2 and various FeCrAl alloys revealed no extensive interaction in all the samples irradiated in the rodlet. Localized defects, namely circular pitting and cracks of a few tents of microns, were observed on the FeCrAl disc surfaces in some of the diffusion couples. These defects might be surface fabrication defects rather than irradiation-induced localized corrosion, but chemical analyses are needed to determine whether accumulation of corrosive fission products are causing enhanced interaction. Both non-destructive and destructive PIEs were collected on a rodlet containing U3Si5 fuel. The rodlet had extremely low fission gas release, similar to previously analyzed U3Si2 rodlets. Gamma tomography revealed thermo-migration of fission product ruthenium toward the center of the fuel pellet. The fuel microstructure resembles the as-fabricated microstructure, showing extensive microcracking related to the phase transition at 450°C. Two types of secondary phases were observed, which could either be due to segregation and precipitation of fission products or to UN impurities from fabrication. Finally, the current status of PIE on other ATF-1 irradiations will be discussed. These irradiations include UN-U3Si5 rodlets, UN-U3Si2 rodlets and an additional test with U3Si2 fuel.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

DOE OSTI2019

Basin-wide gradients in dolomite d26Mg and 87Sr/86Sr identify the structural centre of the Williston Basin as a source of magnesium for dolomitization of the Middle Devonian Winnipegosis Formation

Dolomitized carbonates in the Williston Basin are common below the Middle Devonian Prairie Evaporite Formation, indicating a plausible role for brine-reflux as the source of Mg for dolomitization. However, the Lower Member Winnipegosis Formation, which directly underlies the evaporite sequence, exhibits dolomite 87Sr/86Sr ratios that are higher than expected for Middle Devonian seawater, instead suggesting that the dolomitizing fluids ascended from the deep basin where present-day formation waters are known to be more radiogenic. This interpretation is supported in this study by Mg isotope data showing a basin-scale gradient of increasing d26Mg values from the deep center (–2.03‰) to the edge (–1.34‰) of the Williston Basin along the lower Member Winnipegosis bed. Dolomite favors light isotopes of Mg when it forms, thus driving dolomitizing solutions, and the produced dolomite, towards heavier isotopic compositions in the direction of fluid flow. However, dolomite is also Mg-deficient when it forms, and its initial Mg isotope composition is susceptible to alteration by fluid-rock interactions occurring later in the depositional history of the deposit. We argue that the Mg isotope gradient in the Lower Member Winnipegosis was created by an ascending flow of isotopically light Mg-bearing fluid that differentially altered an earlier formed brine-reflux dolomite, with more alteration occurring deeper in the basin and less towards to edges. Alteration of pre-existing dolomite by an Mg isotope exchange mechanism requires large supplies of Mg. This fact, and the direction of fluid flow point to the underlying crystalline basement as an important Mg source for dolomitization in the Williston Basin. We speculate that ultramafic rocks supplied the Mg when they were hydrothermally altered during an anomalous heating event that affected the Williston Basin in the Late Devonian / Early Carboniferous. Seismic pumping drove these fluids upwards through fracture networks in the crust, and eventually into the deep center of the Williston Basin along vertically-oriented down-to-the-basement faults, pressuring deep aquifers like the Winnipegosis, which resulted in fluids migrating up the stratigraphic dip of the beds towards the edges of the basin. Plotting and contouring the dolomite d26Mg data on a map of the Williston Basin reveals the pathways taken by the migrating fluids. One prominent pathway extending outwards from the center of the basin towards the northeast bears a strong resemblance to petroleum migration fairways deduced for the underlying Late Ordovician Red River Formation. This study demonstrates that Mg isotopic mapping of large dolomite bodies is a useful tool for deciphering fluid flow histories in sedimentary basins and may assist in locating migrated petroleum and Mississippi Valley Type ore deposits.

Magnesium Isotopes, Strontium Isotopes, Dolomitiza↗

DOE OSTI2016

Investigating the Depth of Penetration of Radionuclides in Concrete - 16292

This paper focuses on some chemical and structural aspects for mature concrete structural substrate that functions as a barrier to radioactive material migration. Exposure to radioactive material may be a consequence of surface contact, such as resulting from exposure by spills or leaks. In properly-prepared waste storage areas, undamaged paint and sealant helps to protect the structural cement. However, damaged surfaces, cracks, and relatively porous slab seams can offer a preferred migration pathway for spilled contaminates, including radionuclides. The cost of remediation of concrete structures is directly related to the amount of concrete that must be removed and subsequently disposed. A carefully implemented investigative plan can be utilized to determine how much concrete must be removed and the approximate level of volumetric radiological contamination can be used to estimate disposal costs. Normal concrete pore solution is saturated with calcium hydroxide and also contains sodium and potassium hydroxide; the pH is typically 13 to 14. Contaminants (such as uranium) that hydrolyze in alkaline media that form precipitant phases and tend to deposit at the near-surface. However, more soluble contaminants, such as Cs-137 (as Cs{sup +}) and Tc-99 (as the pertechnetate anion TcO{sub 4}{sup -}) diffuse more rapidly (and deeply) into the porous substrate. Profiling the migration of the water-soluble radionuclides into a cure (hardened) cement surface is a challenge, since water-cooled saws, borers and similar implements are often used to excise and/or slice the core. Alternately, dry mechanical fracturing for excavation of cement may introduce the possibility of material cross-contamination and uncertainty for the average sampling depth for the sample collection event, potentially yielding non-representative samples, and may preclude determination of crack formation that may have been present in the native (non-fractured) substrate. Structural concrete is comprised of cement and pozzolan paste, relatively impervious aggregate (typically stone and/or sand), and rebar (iron reinforcement). Near-surface aggregate may act as a hydrolytic barrier, causing contaminants leached into the pore water to by-pass the impervious grains, causing deeper-than expected penetration, especially if there are grain boundary cracks also present to facilitate migration. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

DOE OSTI2022

ELECTRIC FIELD-ASSISTED DIFFUSION WELDING TO FABRICATE ALLOY 617 COMPACT HEAT EXCHANGERS

Compact heat exchangers are of interest for a number of applications including advanced reactors. Alloy 617 is one of the top candidate materials for the gas-cooled reactor intermediate heat exchanger. Previous endeavors to diffusion weld Alloy 617 utilized hot pressing (HP). It was reported that grain boundary migration across the interface was hindered by extensive precipitation. Bonds of this nature have been observed to reduce the elevated-temperature mechanical properties compared to the wrought-product form. It was hypothesized that the electric current applied during electric-field-assisted sintering (EFAS) can overcome these challenges, resulting in improved diffusion welding (DW). This study investigated DW of Alloy 617 via EFAS. Stacks composed of three sheets that were 20 mm in diameter were welded using EFAS. Specimens were welded with an applied electric current, a pressure of 30 MPa, hold time of 30 min, and temperatures of 1050°C, 1100°C, and 1150°C. DW using HP as the zero-current analog of EFAS was also performed at the most promising EFAS conditions. Results revealed that both the applied electric current and temperature played a key role in precipitation and grain boundary migration in diffusion-welded Alloy 617. Precipitates were observed at the interface of the hot-pressed samples which limited grain boundary migration. Electric current was found to prevent precipitate formation along the interface at 1150°C. The electric current coupled with a temperature of 1150°C during EFAS resulted in significant grain boundary migration across the interface.

36 - MATERIALS SCIENCE↗

DOE OSTI2019

Spatially explicit network analysis reveals multi‐species annual cycle movement patterns of sea ducks

Abstract Conservation of long‐distance migratory species poses unique challenges. Migratory connectivity, that is, the extent to which groupings of individuals at breeding sites are maintained in wintering areas, is frequently used to evaluate population structure and assess use of key habitat areas. However, for species with complex or variable annual cycle movements, this traditional bimodal framework of migratory connectivity may be overly simplistic. Like many other waterfowl, sea ducks often travel to specific pre‐ and post‐breeding sites outside their nesting and wintering areas to prepare for migration by feeding extensively and, in some cases, molting their flight feathers. These additional migrations may play a key role in population structure, but are not included in traditional models of migratory connectivity. Network analysis, which applies graph theory to assess linkages between discrete locations or entities, offers a powerful tool for quantitatively assessing the contributions of different sites used throughout the annual cycle to complex spatial networks. We collected satellite telemetry data on annual cycle movements of 672 individual sea ducks of five species from throughout eastern North America and the Great Lakes. From these data, we constructed a multi‐species network model of migratory patterns and site use over the course of breeding, molting, wintering, and migratory staging. Our results highlight inter‐ and intra‐specific differences in the patterns and complexity of annual cycle movement patterns, including the central importance of staging and molting sites in James Bay, the St. Lawrence River, and southern New England to multi‐species annual cycle habitat linkages, and highlight the value of Long‐tailed Ducks ( Calengula haemalis ) as an umbrella species to represent the movement patterns of multiple sea duck species. We also discuss potential applications of network migration models to conservation prioritization, identification of population units, and integrating different data streams.

54 ENVIRONMENTAL SCIENCES↗

DOE OSTI2015

Radioguided localisation of impalpable breast lesions using 99m-Technetium macroaggregated albumin: Lessons learnt during introduction of a new technique to guide preoperative localisation

Preoperative wire-guided localisation (WGL) of impalpable breast lesions is widely used but can be technically difficult. Risks include wire migration, inaccurate placement, and inadequate surgical margins. Research shows that radioguided occult lesion localisation (ROLL) is quicker, easier, and can improve surgical and cosmetic outcomes. An audited introduction of ROLL was conducted to validate the technique as a feasible alternative to WGL. Fifty patients with single impalpable lesions and biopsy proven malignancy or indeterminate histology underwent WGL followed by intralesional radiopharmaceutical injection of 99m-Technetium macroaggregated albumin. Postprocedural mammography was performed to demonstrate wire position, and scintigraphy to evaluate radiopharmaceutical migration. Lymphoscintigraphy and intraoperative sentinel node biopsy were performed if indicated, followed by lesion localisation and excision using a gamma probe. Specimen imaging was performed, with immediate reexcision for visibly inadequate margins. Accurate localisation was achieved in 86% of patients with ROLL compared to 72% with WGL. All lesions were successfully removed, with clear margins in 71.8% of malignant lesions. Reexcision and intraoperative sentinel node localisation rates were equivalent to preaudit figures for WGL. ROLL was easy to perform and problems were infrequent. Inaccurate radiopharmaceutical placement necessitating WGL occurred in four patients. Minor radiopharmaceutical migration was common, but precluded using ROLL in only two cases. ROLL is effective, simple, inexpensive, and easily learnt; however, preoperative confirmation of correct radiopharmaceutical placement using mammography and the gamma probe is important to help ensure successful lesion removal. Insertion of a backup hookwire is recommended during the initial introduction of ROLL.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

DOE OSTI2017

Analysis of the Human Biohazard of Long-Lived Fission Products and Actinides for BREST-OD-300 Spent Fuel

The aim of this work is to perform a comparative analysis of the biohazard for the population of long-lived fission products and actinides from BREST-OD-300 reactor spent fuel taking account of the time required for different radionuclides to reach Earth’s surface by migrating from a deep disposal site. It was found that the biohazard of fission products is significantly greater than that of actinides for a long time (20000 years according to the accepted scenario of radionuclide migration). The radiological impact on the population is predicted after {sup 99}Tc – radionuclide with high migration capability – reaches the soil surface. {sup 137}Cs and {sup 90}Sr will not appreciably affect the dose load because of their relatively short half-life. In order to maintain radio-ecological balance between long-lived wastes and native uranium during the entire possible time period, it is sufficient to extract from the BREST-OD-300 spent fuel 90% of the technetium and 75% of the americium with 99.9% of the fissile materials extracted.

11 NUCLEAR FUEL CYCLE AND FUEL MATERIALS↗

DOE OSTI2014

Somatic mutational analysis of FAK in breast cancer: A novel gain-of-function mutation due to deletion of exon 33

Highlights: •A novel FAK splicing mutation identified in breast tumor. •FAK-Del33 mutation promotes cell migration and invasion. •FAK-Del33 mutation regulates FAK/Src signal pathway. -- Abstract: Focal adhesion kinase (FAK) regulates cell adhesion, migration, proliferation, and survival. We identified a novel splicing mutant, FAK-Del33 (exon 33 deletion, KF437463), in both breast and thyroid cancers through colony sequencing. Considering the low proportion of mutant transcripts in samples, this mutation was detected by TaqMan-MGB probes based qPCR. In total, three in 21 paired breast tissues were identified with the FAK-Del33 mutation, and no mutations were found in the corresponding normal tissues. When introduced into a breast cell line through lentivirus infection, FAK-Del33 regulated cell motility and migration based on a wound healing assay. We demonstrated that the expression of Tyr397 (main auto-phosphorylation of FAK) was strongly increased in FAK-Del33 overexpressed breast tumor cells compared to wild-type following FAK/Src RTK signaling activation. These results suggest a novel and unique role of the FAK-Del33 mutation in FAK/Src signaling in breast cancer with significant implications for metastatic potential.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2015

Mechanically stimulated bone cells secrete paracrine factors that regulate osteoprogenitor recruitment, proliferation, and differentiation

Bone formation requires the recruitment, proliferation and osteogenic differentiation of mesenchymal progenitors. A potent stimulus driving this process is mechanical loading, yet the signalling mechanisms underpinning this are incompletely understood. The objective of this study was to investigate the role of the mechanically-stimulated osteocyte and osteoblast secretome in coordinating progenitor contributions to bone formation. Initially osteocytes (MLO-Y4) and osteoblasts (MC3T3) were mechanically stimulated for 24hrs and secreted factors within the conditioned media were collected and used to evaluate mesenchymal stem cell (MSC) and osteoblast recruitment, proliferation and osteogenesis. Paracrine factors secreted by mechanically stimulated osteocytes significantly enhanced MSC migration, proliferation and osteogenesis and furthermore significantly increased osteoblast migration and proliferation when compared to factors secreted by statically cultured osteocytes. Secondly, paracrine factors secreted by mechanically stimulated osteoblasts significantly enhanced MSC migration but surprisingly, in contrast to the osteocyte secretome, inhibited MSC proliferation when compared to factors secreted by statically cultured osteoblasts. A similar trend was observed in osteoblasts. This study provides new information on mechanically driven signalling mechanisms in bone and highlights a contrasting secretome between cells at different stages in the bone lineage, furthering our understanding of loading-induced bone formation and indirect biophysical regulation of osteoprogenitors. - Highlights: • Physically stimulated osteocytes secrete factors that regulate osteoprogenitors. • These factors enhance recruitment, proliferation and osteogenic differentiation. • Physically stimulated osteoblasts secrete factors that also regulate progenitors. • These factors enhance recruitment but inhibit proliferation of osteoprogenitors. • This study highlights a contrasting secretome between osteocytes and osteoblasts.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2015

miR-664 negatively regulates PLP2 and promotes cell proliferation and invasion in T-cell acute lymphoblastic leukaemia

MicroRNAs (miRNAs) play important roles in the pathogenesis of many types of cancers by negatively regulating gene expression at posttranscriptional level. However, the role of microRNAs in leukaemia, particularly T-cell acute lymphoblastic leukaemia (T-ALL), has remained elusive. Here, we identified miR-664 and its predicted target gene PLP2 were differentially expressed in T-ALL using bioinformatics methods. In T-ALL cell lines, CCK-8 proliferation assay indicated that the cell proliferation was promoted by miR-664, while miR-664 inhibitor could significantly inhibited the proliferation. Moreover, migration and invasion assay showed that overexpression of miR-664 could significantly promoted the migration and invasion of T-ALL cells, whereas miR-664 inhibitor could reduce cell migration and invasion. luciferase assays confirmed that miR-664 directly bound to the 3'untranslated region of PLP2, and western blotting showed that miR-664 suppressed the expression of PLP2 at the protein levels. This study indicated that miR-664 negatively regulates PLP2 and promotes proliferation and invasion of T-ALL cell lines. Thus, miR-664 may represent a potential therapeutic target for T-ALL intervention. - Highlights: • miR-664 mimics promote the proliferation and invasion of T-ALL cells. • miR-664 inhibitors inhibit the proliferation and invasion of T-ALL cells. • miR-664 targets 3′ UTR of PLP2 in T-ALL cells. • miR-664 negatively regulates PLP2 in T-ALL cells.

60 APPLIED LIFE SCIENCES↗

DOE OSTI2016

GHSR deficiency suppresses neointimal formation in injured mouse arteries

Growth hormone secretagogue receptor (GHSR) is involved in appetite regulation and energy homeostasis. In the present study, we examined the role of GHSR in neointimal formation following vascular injury. In the mouse model of femoral artery wire injury, we found that vessel intima-to-media ratio was significantly reduced in GHSR deficiency (GHSR{sup −/−}) mice compared with that in wild-type mice. Immunohistochemical staining showed that the smooth muscle cell (SMCs) in the neointima were significantly decreased in the injured arteries of GHSR{sup −/−} mice which was associated with decreased SMC proliferation and migration. Furthermore, immunoblotting demonstrated that, in cultured rat aortic SMCs, small interfering RNA-mediated GHSR knockdown suppressed the activation of Akt and ERK1/2 signaling pathway. These findings suggested a novel role of GHSR in neointimal formation likely via promoting the proliferation and migration of SMCs involving Akt and ERK1/2 signaling. Therefore, GHSR may be a potential therapeutic target in restenosis and vascular remodeling. - Highlights: • GHSR deficiency inhibits neointimal formation after vascular injury. • GHSR deficiency suppresses SMCs numbers in vivo. • Knockdown GHSR represses SMCs proliferation and migration in vitro. • Knockdown GHSR inhibited Akt and ERK1/2 phosphorylation in SMCs.

60 APPLIED LIFE SCIENCES↗