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Functional genetic encoding of sulfotyrosine in mammalian cells

Protein tyrosine O -sulfation (PTS) plays a crucial role in extracellular biomolecular interactions that dictate various cellular processes. It also involves in the development of many human diseases. Regardless of recent progress, our current understanding of PTS is still in its infancy. To promote and facilitate relevant studies, a generally applicable method is needed to enable efficient expression of sulfoproteins with defined sulfation sites in live mammalian cells. Here we report the engineering, in vitro biochemical characterization, structural study, and in vivo functional verification of a tyrosyl-tRNA synthetase mutant for the genetic encoding of sulfotyrosine in mammalian cells. We further apply this chemical biology tool to cell-based studies on the role of a sulfation site in the activation of chemokine receptor CXCR4 by its ligand. Our work will not only facilitate cellular studies of PTS, but also paves the way for economical production of sulfated proteins as therapeutic agents in mammalian systems.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Structure of an EIIC sugar transporter trapped in an inward-facing conformation

The phosphoenolpyruvate-dependent phosphotransferase system (PTS) transports sugar into bacteria and phosphorylates the sugar for metabolic consumption. The PTS is important for the survival of bacteria and thus a potential target for antibiotics, but its mechanism of sugar uptake and phosphorylation remains unclear. The PTS is composed of multiple proteins, and the membrane-embedded Enzyme IIC (EIIC) component transports sugars across the membrane. Crystal structures of two members of the glucose superfamily of EIICs, bcChbC and bcMalT, were solved in the inward-facing and outward-facing conformations, and the structures suggest that sugar translocation could be achieved by movement of a structured domain that contains the sugar-binding site. However, different conformations have not been captured on the same transporter to allow precise description of the conformational changes. Here we present a crystal structure of bcMalT trapped in an inward-facing conformation by a mercury ion that bridges two strategically placed cysteine residues. The structure allows direct comparison of the outward- and inward-facing conformations and reveals a large rigid-body motion of the sugar-binding domain and other conformational changes that accompany the rigid-body motion. All-atom molecular dynamics simulations show that the inward-facing structure is stable with or without the cross-linking. Here, the conformational changes were further validated by single-molecule Föster resonance energy transfer (smFRET). Combined, these results establish the elevator-type mechanism of transport in the glucose superfamily of EIIC transporters.

59 BASIC BIOLOGICAL SCIENCES↗

Pulsar-Calibrated Timing Source for Synchronized Sampling

The Global Positioning System (GPS) is critical to the real-time synchronized sampling of phasor measurement units (PMUs). Unfortunately, GPS signals are occasionally unstable due to several factors such as weak satellite signal and GPS spoofing, thereby leaving the PMU with a degraded sampling performance. In this letter, a novel Pulsar-calibrated Timing Source (PTS) is proposed as the alternative timing source for synchronized sampling. Further, the PTS can generate the timing signal with a 1 μs drift error within 91 holdover minutes to ensure continuity of sampling accuracy. Experimental tests are conducted and the results reveal the reliability and accuracy of the PTS for PMU synchronization.

42 ENGINEERING↗

Study of serious adverse drug reactions using FDA-approved drug labeling and MedDRA

Background Adverse Drug Reactions (ADRs) are of great public health concern. FDA-approved drug labeling summarizes ADRs of a drug product mainly in three sections, i.e., Boxed Warning (BW), Warnings and Precautions (WP), and Adverse Reactions (AR), where the severity of ADRs are intended to decrease in the order of BW > WP > AR. Several reported studies have extracted ADRs from labeling documents, but most, if not all, did not discriminate the severity of the ADRs by the different labeling sections. Such a practice could overstate or underestimate the impact of certain ADRs to the public health. In this study, we applied the Medical Dictionary for Regulatory Activities (MedDRA) to drug labeling and systematically analyzed and compared the ADRs from the three labeling sections with a specific emphasis on analyzing serious ADRs presented in BW, which is of most drug safety concern. Results This study investigated New Drug Application (NDA) labeling documents for 1164 single-ingredient drugs using Oracle Text search to extract MedDRA terms. We found that only a small portion of MedDRA Preferred Terms (PTs), 3819 out of 21,920 or 17.42%, were observed in a whole set of documents. In detail, 466/3819 (12.0%) PTs were in BW, 2023/3819 (53.0%) were in WP, and 2961/3819 (77.5%) were in AR sections. We also found a higher overlap of top 20 occurring BW PTs with WP sections compared to AR sections. Within the MedDRA System Organ Class levels, serious ADRs (sADRs) from BW were prevalent in Nervous System disorders and Vascular disorders. A Hierarchical Cluster Analysis (HCA) revealed that drugs within the same therapeutic category shared the same ADR patterns in BW (e.g., nervous system drug class is highly associated with drug abuse terms such as dependence, substance abuse, and respiratory depression). Conclusions This study demonstrated that combining MedDRA standard terminologies with data mining techniques facilitated computer-aided ADR analysis of drug labeling. We also highlighted the importance of labeling sections that differ in seriousness and application in drug safety. Using sADRs primarily related to BW sections, we illustrated a prototype approach for computer-aided ADR monitoring and studies which can be applied to other public health documents.

59 BASIC BIOLOGICAL SCIENCES↗

Predicting Outcome in Mammary Phyllodes Tumors: Relevance of Clinicopathological Features

Background: Phyllodes tumors (PTs) of the breast are uncommon fibroepithelial neoplasms. Most behave in a benign fashion but they also have the potential to recur locally or to metastasize. Methods: In the current study involving 290 PTs (181 benign, 76 borderline, and 33 malignant) from three hospitals over an 11-year period, we assessed the relationship between histologic parameters (including histologic features affecting grade and surgical margin status), postoperative adjuvant treatment, and local recurrences and distant metastases. Results: An involved surgical margin was the only factor associated with increased risk of local recurrences (hazard ratio [HR] 4.673, p = 0.003), but not for distant metastases. For local recurrences, a wider margin did not confer additional benefits. None of the histologic factors were predictive for local recurrences. In contrast, distant metastases were correlated with histologic parameters, particularly an infiltrative border (HR 10.935, p = 0.012) and the presence of necrosis (HR 15.311, p = 0.007). In this series, all local recurrences were found in patients without radiotherapy, regardless of surgical margin status. Conclusion: A negative surgical margin is mandatory for the effective local control of PT recurrence, and a minimal margin clearance may be sufficient. For distant metastases, the inherent characteristics of PTs are important, thus it may be prudent to evaluate additional histologic features, including necrosis, for patients’ prognostication.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

Summary of NDC Capacity Building Workshop and Regional Seismic Travel Time (RSTT) in combination with Data Sharing and Integration Training

The NNSA Seismic Cooperation Program (SCP) sponsored Stephen Myers (LLNL), Michael Begnaud (LANL), Brian Young (SNL) and Istvan Bondar (Research Center for Astronomy and Earth Sciences, Hungary) to serve as a presenters/trainers at the “NDC Capacity Building Workshop and Regional Seismic Travel Time (RSTT) in combination with Data Sharing and Integration Training” September 4-8 2022 in Muscat, Oman (See Appendix A for the agenda). The workshop and training (workshop from here forward) was organized by the Comprehensive Nuclear-Test-Ban Treaty Organization (CTBTO) Provisional Technical Secretariat (PTS). The first half of the week was devoted to NDC workshop activities, and the second half was devoted to RSTT training. Fifty-five participants from 27 countries and the CTBTO-PTS attended the 5-day workshop (See Appendix B for list of participants and countries of origin). Presentations from the PTS described the International Monitoring System (IMS), International Data Centre (IDC) products, and metrics of regional data utilization. Contributed presentations from each country’s scientists included descriptions of regional and national networks, methods of data analysis, and needs for material and technical assistance. Training included an overview of the RSTT method and instruction on how to locate seismic events with the iLoc program, which utilizes RSTT travel times to reduce bias in event location estimates. Methods of seismic tomography and the need for a high-quality tomographic set, including seismological “ground truth”, were emphasized. Seismological “ground truth” or “GT” is a term that has come to mean both events with known location and events with well-characterized locations that are estimated using seismological data, typically with epicenter accuracy of 5 km or better. Notably, the instructional platform has migrated from UNIX shell scripts to Jupyter Notebooks. Jupyter Notebooks have the advantage being more visually intuitive, including display of graphics within the notebook. Each notebook includes every processing step that participants need to reproduce the entire exercise.

58 GEOSCIENCES↗

Operational evaluation of a proppeller test stand in the quiet flow facility at Langley Research Center

Operational proof tests of a propeller test stand (PTS) in a quiet flow facility (QFF) are presented. The PTS is an experimental test bed for acoustic propeller research in the quiet flow environment of the QFF. These proof tests validate thrust and torque predictions, examine the repeatability of measurements on the PTS, and determine the effect of applying artificial roughness to the propeller blades. Since a thrusting propeller causes an open jet to contract, the potential flow core was surveyed to examine the magnitude of the contraction. These measurements are compared with predicted values. The predictions are used to determine operational limitations for testing a given propeller design in the QFF.

Block, P. J. W.↗

A new Antarctic meteorite with chromite, orthopyroxene and metal with reference to a formation model of S asteroids

Mineralogy of a unique meteorite from Antarctica, EET84302 and the related primitive achondrites, was studied to gain better understanding on relationship between S asteroids and this meteorite group. EET84302 consists of metal-rich (M) region with metal filling interstices of silicate grains of orthopyroxene (Opx), olivine Fa8, plagioclase An24, and augite (Aug), chromite-rich (C) region with major chromite and Opx, and silicate-rich (S) region with mainly Opx. This is the first example of chromite-rich achondrite. Distribution of three areas (C, M, S) are in line with a proposed model of primitive achondrites and S asteroids. Since reflectance spectra of some members of primitive achondrites combined with those of iron meteorites, resemble those of the S asteroids, there is an expectation that S asteroids are modified products of chondrites. The trends of the variations in their mineral assemblages are in line with those of the S asteroids. Hiroi proposed a stony-iron model of S asteroids, which assumes that a few domains of various primitive achondrites are embedded in iron matrix. Polished thin sections (PTS) of EET84302,19 and ALH81187,16 were supplied from Meteorite Working Group (MWG). A previously studied PTS of EET84302 was similar in both texture and mineralogy to silicate inclusions in some iron meteorites. PTS EET84302,19 was studied by electron probe microanalyzer (EPMA) and scanning electron microscope (SEM). The reflectance spectra of EET84302 showed a weathering problem and was not used to match with those of S asteroids.

Takeda, Hiroshi↗

Lab-on-a-Chip: From Astrobiology to the International Space Station

The continual and long-term habitation of enclosed environments, such as Antarctic stations, nuclear submarines and space stations, raises unique engineering, medical and operational challenges. There is no easy way out and no easy way to get supplies in. This situation elevates the importance of monitoring technology that can rapidly detect events within the habitat that affect crew safety such as fire, release of toxic chemicals and hazardous microorganisms. Traditional methods to monitor microorganisms on the International Space Station (ISS) have consisted of culturing samples for 3-5 days and eventual sample return to Earth. To augment these culture methods with new, rapid molecular techniques, we developed the Lab-on-a-Chip Application Development - Portable Test System (LOCAD-PTS). The system consists of a hand-held spectrophotometer, a series of interchangeable cartridges and a surface sampling/dilution kit that enables crew to collect samples and detect a range of biological molecules, all within 15 minutes. LOCAD-PTS was launched to the ISS aboard Space Shuttle Discovery in December 2006, where it was operated for the first time during March-May 2007. The surfaces of five separate sites in the US Lab and Node 1 of ISS were analyzed for endotoxin, using cartridges that employ the Limulus Amebocyte Lysate (LAL) assay; results of these tests will be presented. LOCAD-PTS will remain permanently onboard ISS with new cartridges scheduled for launch in February and October of 2008 for the detection of fungi (Beta-glucan) and Gram-positive bacteria (lipoteichoic acid), respectively.

Maule, Jake↗

Rapid Monitoring of Bacteria and Fungi aboard the International Space Station (ISS)

Microorganisms within spacecraft have traditionally been monitored with culture-based techniques. These techniques involve growth of environmental samples (cabin water, air or surfaces) on agar-type media for several days, followed by visualization of resulting colonies or return of samples to Earth for ground-based analysis. Data obtained over the past 4 decades have enhanced our understanding of the microbial ecology within space stations. However, the approach has been limited by the following factors: i) Many microorganisms (estimated > 95%) in the environment cannot grow on conventional growth media; ii) Significant time lags (3-5 days for incubation and up to several months to return samples to ground); iii) Condensation in contact slides hinders colony counting by crew; and iv) Growth of potentially harmful microorganisms, which must then be disposed of safely. This report describes the operation of a new culture-independent technique onboard the ISS for rapid analysis (within minutes) of endotoxin and beta-1, 3-glucan, found in the cell walls of gramnegative bacteria and fungi, respectively. The technique involves analysis of environmental samples with the Limulus Amebocyte Lysate (LAL) assay in a handheld device, known as the Lab-On-a-Chip Application Development Portable Test System (LOCAD-PTS). LOCADPTS was launched to the ISS in December 2006, and here we present data obtained from Mach 2007 until the present day. These data include a comparative study between LOCADPTS analysis and existing culture-based methods; and an exploratory survey of surface endotoxin and beta-1, 3-glucan throughout the ISS. While a general correlation between LOCAD-PTS and traditional culture-based methods should not be expected, we will suggest new requirements for microbial monitoring based upon culture-independent parameters measured by LOCAD-PTS.

Gunter, D.↗

Enzyme-Cascade Analysis of the Rio Tinto Subsurface Environment: A Biosensor Experiment

The Portable Test System (PTS), designed & developed by Charles Rivers Laboratories, Inc. (Charleston, SC) is a portable instrument that was designed to perform analysis of enzymatic assays related to rapid assessment of microbial contamination (Wainwright, 2003). The enzymatic cascade of Limulus Amebocyte Lysate (LAL) is known to be one of the most sensitive techniques available for microbial detection, enabling the PTS to be evaluated as a potential life detection instrument for in situ Astrobiology missions. In the summer of 2003 the system was tested as a part of the Mars Astrobiology Research and Technology Experiment (MARTE) ground truth science campaign in the Rio Tinto Analogue environment near Nerva, Spain. The preliminary results show that the PTS analysis correlates well with the contamination control tests and the more traditional lab-based biological assays performed during the MARTE field mission. Further work will be conducted on this research during a second field campaign in 2004 and a technology demonstration of a prototype instrument that includes autonomous sample preparation will occur in 2005.

McKay, David S.↗

Virtual Microscope Views of the Apollo 11 and 12 Lunar Samples

The Apollo virtual microscope is a means of viewing, over the Internet, polished thin sections of every rock in the Apollo lunar sample collections via software, duplicating many of the functions of a petrological microscope, is described. Images from the Apollo 11 and 12 missions may be viewed at: www.virtualmicroscope.org/content/apollo. Introduction: During the six NASA missions to the Moon from 1969-72 a total of 382 kilograms of rocks and soils, often referred to as "the legacy of Apollo", were collected and returned to Earth. A unique collection of polished thin sections (PTSs) was made from over 400 rocks by the Lunar Sample Curatorial Facility at the Johnson Spacecraft Center (JSC), Houston. These materials have been available for loan to approved PIs but of course they can't be simultaneously investigated by several researchers unless they are co-located or the sample is passed back and forward between them by mail/hand carrying which is inefficient and very risky for irreplaceable material. When The Open University (OU), the world's largest Distance Learning Higher Education Establishment found itself facing a comparable problem (how to supply thousands of undergraduate students with an interactive petrological microscope and a personal set of thin sections), it decided to develop a software tool called the Virtual Microscope (VM). As a result it is now able to make the unique and precious collection of Apollo specimens universally available as a resource for concurrent study by anybody in the world's Earth and Planetary Sciences community. Herein, we describe the first steps of a collaborative project between OU and the Johnson Space Center (JSC) Curatorial Facility to record a PTS for every lunar rock, beginning with those collected by the Apollo 11 and 12 missions. Method: Production of a virtual microscope dedicated to a particular theme divides into four main parts - photography, image processing, building and assembly of virtual microscope components, and publication on a website. Two large research quality microscopes are used to collect all the images required for a virtual microscope. The first is part of an integrated package that utilizes Leica PowerMosaic software and a motorised XYZ stage to generate large area mosaics. It includes a fast acquisition camera and depending on the PTS size normally is used to produce seamless mosaic images consisting of 100-500 individual photographs. If the sample is suitable, three mosaics of each sample are recorded - plane polarised light, between crossed polars and reflected light. In order for the VM to be a true petrological microscope it is necessary to recreate the features of a rotating stage and perform observations using filters to produce polarised light. Thus the petrological VM includes the capability of seeing changes in optical properties (pleochroism and birefringence) during rotation allowing mineral identification. The second microscope in the system provides the functions of the rotating stage. To this microscope we have added a robotically controlled motor to acquire seventy-two images (5 degree intervals) in plane polarised light and between crossed polars. To process the images acquired from the two microscopes involves a combination of proprietary software (Photoshop) and our own in-house code. The final stage involves assembling all the components in an HTML5 environment. Pathfinder investigations: We have undertaken a number of pilot studies to demonstrate the efficacy of the petrological microscope with lunar samples. The first was to make available on-line images collected from the Educational Package of Apollo samples provided by NASA to the UK STFC (Science and Technical Facilities Council) for loan as educational material e.g. for schools. The real PTSs of the samples are now no longer sent out to schools removing the risks associated with transport, accidental breakage and eliminating the possibility of loss. The availability of lunar sample VM-related material was further extended to include twenty-eight specimens from all of the Apollo missions. Some of these samples were made more generally available through an ibook entitled "Moon Rocks: an introduction to the Geology of the Moon," free from the Apple Bookstore. Research possibilities: Although the Virtual Microscope was originally conceived as a teaching aid and was later recognised as a means of public outreach and engagement, we now realize that it also has enormous potential as a high level research tool. Following discussions with the JSC Curators we have received Curation and Analysis Planning Team for Extraterrestrial Materials (CAPTEM) permission to embark on a programme of digitizing the entire lunar sample PTS collection for all three of the above purposes. By the time of the 47th Lunar and Planetary Science Conference (LPSC) we will have completed 81 rocks collected during the Apollo 11 and 12 missions and the data, with cross-links to the Lunar Sample Compendium will go live on the Web at the 47th LPSC. The VM images of the Apollo 11 (41 VM images) and 12 (40 VM images) missions can be viewed at: http:/www.virtualmicroscope.org/content/apollo. The lunar sample VM will enable large numbers of skilled/unskilled microscopists (professional and amateur researchers, educators and students, enthusiasts and the simply curious non-scientists) to share the information from a single sample. It will mean that all the PTSs already cut, even historical ones, could be available for new joint investigations or private study. The scientific return from the collection will increase exponentially as a result of further debate and discussion. Simultaneously the VM will remove the need for making unnecessary multiple samplings, avoid consignment of delicate/breakable specimens (all of which are priceless) to insecure mail/courier services and reduce direct labour and indirect costs, travel budgets and unproductive travelling time necessary for co-location of collaborating researchers. For the future we have already recognized further potential for virtual technology. There is nothing that a petrologist likes more than to see the original rock as a hand specimen. It is entirely possible to recreate virtual hand specimens with 3-D hard and software, already developed for viewing fossils, located within the Curatorial Facility, http://curator.jsc.nasa.gov/lunar/lsc/index.cfm.

Gibson, E. K.↗

Coherence Analysis of the Space Launch System using Unsteady Pressure Sensitive Paint

Transonic buffet during atmospheric ascent is a major source of unsteady loading on launch vehicles that, in the past, has led to structural failures. Thus, determining accurate buffet forcing functions (BFFs) to properly predict the vehicle response to buffet is of vital importance in launch vehicle design. The state of the art for obtaining the BFFs relies on wind-tunnel tests where the fluctuating pressures are measured by pressure transducers (PTs) at sparse locations on a rigid, geometrically-scaled buffet model. To compute the BFFs, the outer mold line (OML) of the vehicle is mapped onto contiguous panels centered at the location of the PTs and the measured fluctuating pressures are integrated over the panels’ areas. To mitigate conservatism due to the assumption that the measured buffet pressures act in phase across each panel, coherence factors are applied, effectively reducing the integration areas and, therefore, the buffet forces. For coherence factors to provide the proper level of BFF attenuation, accurate knowledge of the spatial and temporal correlation of the buffet pressures is paramount. Unfortunately, even with hundreds of unsteady pressure transducers instrumenting the models, compromises must be made on the spatial resolution of the PTs. Typically, the PT layout aims at resolving the pressure correlation along the longitudinal axis of the vehicle, assuming that coherent structures propagate mostly in the longitudinal direction. As a result, the distribution of azimuthal/cross-stream correlation is not well known and its impact on the estimated BFFs is often neglected. To fill this and other knowledge gaps, extremely high spatial-resolution uPSP data were collected for three different production-design configurations of the Space Launch System in the NASA Ames Research Center Unitary Plan Wind Tunnel 11-Foot Transonic Wind Tunnel. Specifically, two cargo configurations, the Block 1 and Block 1B, and one crew configuration, the Block 1B crew, were surveyed at resolutions ranging from 600,000 to over 1 million uPSP measurement locations. To shed light on the temporal and spectral behavior of are presented. Several OML regions and flow features of interest are investigated, from theexpansion/shock on the Orion Multi-Purpose Crew Vehicle, to the terminal shock environment on the core stage, and the Strouhal shedding behind the boosters forward attach. The sensitivity of these environments to the vehicle attitude is examined. Furthermore, for selected panels,coherence factors that accurately capture the azimuthal coherence distribution of the buffetpressures are derived and their impact on the estimated BFFs is discussed. Finally, distributions of the local convection velocity and cross spectrum phase are presented.

buffet↗

NASA's Exploration and In-Space Services (NExIS) Division OSAM-1 Propellant Transfer Subsystem Progress 2020

National Aeronautics and Space Administration (NASA)’s Exploration and In-Space Services (NExIS) Division of Goddard Space Flight Center has been developing technology for the On-orbit Servicing, Assembly and Manufacturing Mission 1 (OSAM-1) to robotically refuel heritage and new satellites on-orbit. OSAM-1, formerly known as Restore-L, successfully passed an important NASA milestone called Key Decision Point-C (KDP-C), receiving agency-level approval for its implementation. The decision point also establishes the mission’s official schedule and budget. The OSAM-1 spacecraft, Servicing Payload and the Space Infrastructure Dexterous Robot (SPIDER) payload will refuel a satellite in space, assemble a communications antenna and manufacture a beam. By demonstrating these capabilities, the mission is advancing never-before tested technologies for use in future missions (by NASA and other government organizations and private industry). The mission is funded by the Technology Demonstration Missions program within NASA’s Space Technology Mission Directorate. This paper covers a review of servicing extensibility and critical technologies that are being developed within NExIS with a focus on the fluid transfer refueling technology within the framework of the Propellant Transfer Subsystem (PTS). An overview of the planned initial technology demonstration servicing mission via the OSAM-1 Space Vehicle is provided as an extension of the technology development progress reported in 20181, and 20192. The general objectives, challenges, and key technologies are presented as an introduction to the context of the OSAM-1 mission, and a precursor to the OSAM-1 PTS specific development status. Development and progress of the Hose Management Assembly (HMA) and Propellant Transfer Assembly (PTA) are discussed. HMA risk reduction test results including those of thermal vacuum testing are presented. Important analytical results are discussed and progress of drawings and procedures developed for the fabrication and testing phase are shown. A summary of the PTS overall verification status and design activities prepared for the critical design peer review are presented. Procurement progress is shown with status on long, medium, and short term efforts to acquire the hardware required to support the mission objectives. Technology development, challenges, and testing status are discussed with particular regard to the OSAM-1 specific key assemblies including the PTA and HMA along with the overall integrated flight mockup of the spacecraft to client fluid transfer testing conducted in the servicing testbeds. The paper concludes with key milestones leading to the goal of on-orbit refueling to be demonstrated in 2024. The overall mission Launch Readiness Date (LRD) has been realigned to accommodate budget profiles and incorporation of additional on-orbit assembly and manufacturing demonstration efforts.

Propellant transfer, satellite servicing↗

Comparative studies of glycolytic pathways and channeling under in vitro and in vivo modes

This study constructed cell‐free glycolytic enzyme systems and compared them to their in vivo functions in Escherichia coli . Under in vitro conditions, flux regulation followed enzyme concentrations and kinetics. In E. coli, only one of the isozymes of phosphofructokinase (PfkA) and fructose‐bisphosphate aldolase (FbaA) facilitate Embden‐Meyerhof‐Parnas (EMP) flux, but under in vitro assays, these isozymes were interchangeable. Additionally, in vitro introduction of the Entner–Doudoroff (ED) pathway improved glycolysis rates, while in vivo overexpression of the ED pathway could not capture significant flux unless its phosphotransferase system (PTS) was knocked out. Lastly, in vivo dynamic 13 C‐experiments revealed that the labeling order of EMP pathway intermediates was not strictly cascade, indicating intracellular metabolites were not well mixed. These enigmatic observations cannot be fully explained by thermodynamics or substrate level regulations. This article supports the long‐time conjecture that EMP enzymes are channeled, and the PTS may be an anchor point to initiate enzyme assemblies. © 2018 American Institute of Chemical Engineers AIChE J , 65: 483–490, 2019

Abernathy, Mary H.↗

Detection of phosphate transporter genes from arbuscular mycorrhizal fungi in mature tree roots under experimental soil pH manipulation

We present the majority of terrestrial plant roots are colonized by arbuscular mycorrhizal (AM) fungi that, in exchange for carbon, provide plants with enhanced nutrient uptake — most notably inorganic phosphate (P i ). To mediate the uptake of Pi from the soil, AM fungi possess high affinity inorganic phosphate transporters (PTs). Under laboratory conditions, P i concentrations have been shown to regulate AM fungal-specific PT gene expression. The relationship between PT expression and P i in the field remains unexplored. Here we quantify AM fungal-specific PTs from maple tree roots in situ. In an effort to limit edaphic parameters, root samples were collected from manipulated forested plots that had elevated soil P i availability, either through direct P i application or elevating pH to lower exchangeable aluminum. The aim of the study was to examine AM fungal-specific PT gene expression both prior to and following soil P i amendment; however, a direct correlation between soil P i concentration and PT gene expression was not observed. PT transcripts were detected to a greater extent under elevated pH and, while our results are confounded by an overall low detection of PT genes (23 % of all samples collected), our findings raise interesting questions regarding the role of soil pH on PT function. In conclusion, our study is a first step in understanding how edaphic properties influence PT expression and plant P acquisition in mature tree roots.

54 ENVIRONMENTAL SCIENCES↗

An Evaluation of “Drug Ineffective” Postmarketing Reports in Drug Safety Surveillance

Introduction: The most commonly reported adverse event, based on frequency of Medical Dictionary for Regulatory Activities (MedDRA) preferred terms (PTs), in the US FDA Adverse Event Reporting System (FAERS) database is ‘‘drug ineffective’’ (DI). This study aimed to describe the DI reports and provide data to support recommendations on how to best evaluate these reports. Methods: We characterized all FAERS reports coded with the MedDRA PT ‘‘drug ineffective’’ received between 1 September 2012 and 31 August 2016 using all other FAERS reports as a comparator. Additionally, we conducted a manual evaluation to identify informative data elements in the report narratives. Results: During the study period, 247,513 (6.4% of all FAERS reports) DI reports were entered in FAERS. Compared with non-DI reports, DI reports were more likely to be reported by consumers (69.8 vs. 48.1%) and less likely to report a serious outcome (26.2 vs. 56.3%). Most DI reports (88%) were from the USA. Manual evaluation of 552 sample US reports identified 43 reports (7.8%) deemed ‘‘useful’’; a higher proportion of ‘‘useful’’ reports provided a batch or lot number (39.5 vs. 17.2%) and were coded with additional PTs beyond ‘‘drug ineffective’’ (83.7 vs. 59.2%), the most frequent of which were ‘‘product quality issue’’ (23.3%) and ‘‘product substitution issue’’ (18.6%). Conclusions: DI was the most frequently reported adverse event in the FAERS database; however, the yield from these reports in terms of usefulness from a pharmacovigilance perspective was low. Efficient strategies are needed to identify which DI reports are more likely to contain useful information.

60 APPLIED LIFE SCIENCES↗

Decoding anomalous grain growth at room temperature during pressure-induced phase transformations

Significant grain growth is observed during the high-pressure phase transformations (PTs) at room temperature in various materials. The main focus here is grain growth from a few hundred nanometers to 10 μm within an hour during α → ω PT in Zr. No existing theory explains this phenomenon since without PT, Zr nanocrystals do not grow at room temperature even for up to 10 years. Here, in this study, a multistep mechanism for the grain growth during α → ω PT in Zr is suggested. Phase interfaces (PI) and grain boundaries (GBs) coincide and move together as a combined PI-GBs under the action of the combined thermodynamic driving force. Such a combined motion changes the diffusional grain growth mechanism to the transformational one and the martensitic mechanism of PT to a reconstructive one via an intermediate disordered phase. The primary condition is that the GB energy of the ω phase is smaller than that of the α phase, which promotes the nucleation of ω-Zr and is consistent with the absence of the reverse PT and reduction in the PT pressure with reducing grain size. Several intermediate steps for such motion are suggested and justified kinetically. Nonhydrostatic stresses due to volume reduction in the growing ω grain promote continuous growth of the existing ω grain instead of a new nucleation at other GBs. In situ synchrotron Laue diffraction experiments confirm the main predictions of the theory. The suggested mechanism provides a new insight into synergistic interaction between PTs and microstructure evolution.

anomalous grain growth during phase transformation↗