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At least 109 records · Page 6

Identification of a specific exporter that enables high production of aconitic acid in Aspergillus pseudoterreus

Aconitic acid is an unsaturated tricarboxylic acid that is attractive for its potential use in the manufacture of biodegradable and biocompatible polymers, plasticizers, and surfactants. Previously Aspergillus pseudoterreus was engineered as a platform to produce aconitic acid by deleting the cadA (cis-aconitic acid decarboxylase) gene in the itaconic acid biosynthetic pathway. In this study aconitic acid transporter gene (aexA) was identified using comparative global discovery proteomics analysis between the wild-type and cadA deletion strains. Deletion of aexA almost eliminated aconitic acid secretion, while its overexpression led to a significant increase in aconitic acid production. Transportation of aconitic acid across the plasma membrane is a key limiting step. In vitro proteoliposome transport assay further validated AexA’s function and its substrate specificity. This research provides new approaches to efficiently pinpoint and characterize exporters of fungal organic acids and accelerate the metabolic engineering to improve secretion capability and lower cost for bioproduction.

59 BASIC BIOLOGICAL SCIENCES↗

Final Technical Report - Autothermal Styrene Manufacturing Process with Net Export of Energy

The overall objectives of the project were to: (a) develop an economically competitive processing technology for styrene monomer (SM) that would reduce process energy requirements by a minimum 25% relative to those of conventional technology while achieving a minimum 10% ROI; and (b) advance the technology towards commercial readiness. This technology is referred to as OMT (Oxymethylation of Toluene). The unique energy savings feature of the OMT technology would be replacement of the conventional benzene and ethylene feedstocks with toluene, methane in natural gas and air or oxygen, the latter of which have much lower specific energy of production values. As an oxidative technology, OMT is a net energy exporter rather than a net energy consumer like the conventional ethylbenzene/styrene (EB/SM) process. OMT plants would ultimately reduce the cost of styrene monomer which in turn will decrease the costs of polystyrene making it perhaps more cost competitive with competing polymers such as polypropylene.

03 NATURAL GAS↗

Russian Support for Innovation and Export Growth

State support for innovation is considered for the example of research into the commercialization of new manufacturing products. Existing models for the support and stimulation of innovations are presented, and the development of state programs capable of stimulating export expansion on the basis of innovations in mechanical engineering is proposed.

29 ENERGY PLANNING, POLICY, AND ECONOMY↗

Rating Correlations Between Customs Codes and Export Control Lists: Assessing the Needs and Challenges

Correlation tables are the linchpins between the customs codes used to classify commodities in international trade and the control lists used for strategic trade control (STC) purposes. While understanding the customs classification system can help the STC community better understand strategic trade flows, better identify which trade operations require permits, and more effectively detect illegal exports, the two systems are different in scope, philosophy, content, and objectives. Many indications point to the limitations of these correlation tables, and it is important to understand the nature of the limitations and the complex underlying reasons to conceive possible improvements. As part of its Strategic Trade and Supply Chain Analytics Initiative, Argonne National Laboratory supported a study of a subset of the European Union’s TARIC correlation table. The study included development of a methodology and an approach to rating the quality and relevance of individual correlations. The study was intended as a first step to engage the STC community in deflections and initiatives to improve the conception and use of correlations, and its conclusions illustrate the scope and complex nature of the challenges to overcome. This paper presents the two classification systems, analyzes the needs for correlation tables and the complex challenges associated with them, summarizes key findings, and proposes possible ways forward.

commodity classification↗

Life Cycle Analysis of Coal Exports from the Powder River Basin

This presentation was given at the 2016 Carbon Capture, Utilization, and Storage Conference in June 2016, and provided the complete technical details behind and the full life cycle impacts of exporting coal from the Powder River Basin in the U.S. to electricity markets in Asia. Key takeaways are the relative large contribution of the transportation of the coal when traveling distance.

Climate Change↗

Importance of lateral flux and its percolation depth on organic carbon export in Arctic tundra soil: Implications from a soil leaching experiment

Temperature rise in the Arctic is causing deepening of active layers and resulting in the mobilization of deep permafrost dissolved organic matter (DOM). However, the mechanisms of DOM mobilization from Arctic soils, especially upper soil horizons which are drained most frequently through a year, are poorly understood. Here, we conducted a short-term leaching experiment on surface and deep organic active layer soils, from the Yukon River basin, to examine the effects of DOM transport on bulk and molecular characteristics. We report our data showed a net release of DOM from surface soils equal to an average of 5% of soil carbon. Conversely, deep soils percolated with surface leachates retained up to 27% of bulk DOM-while releasing fluorescent components (up to 107%), indicating selective release of aromatic components (e.g. lignin, tannin), while retaining non-chromophoric components, as supported by spectrofluorometric and ultra high resolution mass spectroscopic techniques. Our findings highlight the importance of the lateral flux of DOM on ecosystem carbon balance as well as processing of DOM transport through organic active layer soils en route to rivers and streams. This work also suggests the potential role of leachate export as an important mechanism of C losses from Arctic soils, in comparison with the more traditional pathway from soil to atmosphere in a warming Arctic.

54 ENVIRONMENTAL SCIENCES↗

RNA interference regulates the cell cycle checkpoint through the RNA export factor, Ptr1, in fission yeast

Highlights: Black-Right-Pointing-Pointer RNAi is linked to the cell cycle checkpoint in fission yeast. Black-Right-Pointing-Pointer Ptr1 co-purifies with Ago1. Black-Right-Pointing-Pointer The ptr1-1 mutation impairs the checkpoint but does not affect gene silencing. Black-Right-Pointing-Pointer ago1{sup +} and ptr1{sup +} regulate the cell cycle checkpoint via the same pathway. Black-Right-Pointing-Pointer Mutations in ago1{sup +} and ptr1{sup +} lead to the nuclear accumulation of poly(A){sup +} RNAs. -- Abstract: Ago1, an effector protein of RNA interference (RNAi), regulates heterochromatin silencing and cell cycle arrest in fission yeast. However, the mechanism by which Ago1 controls cell cycle checkpoint following hydroxyurea (HU) treatment has not been elucidated. In this study, we show that Ago1 and other RNAi factors control cell cycle checkpoint following HU treatment via a mechanism independent of silencing. While silencing requires dcr1{sup +}, the overexpression of ago1{sup +} alleviated the cell cycle defect in dcr1{Delta}. Ago1 interacted with the mRNA export factor, Ptr1. The ptr1-1 mutation impaired cell cycle checkpoint but gene silencing was unaffected. Genetic analysis revealed that the regulation of cell cycle checkpoint by ago1{sup +} is dependent on ptr1{sup +}. Nuclear accumulation of poly(A){sup +} RNAs was detected in mutants of ago1{sup +} and ptr1{sup +}, suggesting there is a functional link between the cell cycle checkpoint and RNAi-mediated RNA quality control.

60 APPLIED LIFE SCIENCES↗

Erythropoietin and carbamylated erythropoietin promote histone deacetylase 5 phosphorylation and nuclear export in rat hippocampal neurons

Erythropoietin (EPO) produces neurotrophic effects in animal model of neurodegeneration. However, clinical use of EPO is limited due to thrombotic risk. Carbamylated EPO (cEPO), devoid of thrombotic risk, has been proposed as a novel neuroprotective and neurotrophic agent although the molecular mechanisms of cEPO remain incomplete. Here, we show a previously unidentified role of histone deacetylase 5 (HDAC5) in the actions of EPO and cEPO. EPO and cEPO regulate the HDAC5 phosphorylation at two critical sites, Ser259 and Ser498 through a protein kinase D (PKD) dependent pathway. In addition, EPO and cEPO rapidly stimulates nuclear export of HDAC5 in rat hippocampal neurons which expressing HDAC5-GFP. Consequently, EPO and cEPO enhanced the myocyte enhancer factor-2 (MEF2) target gene expression. Taken together, our results reveal that EPO and cEPO mediate MEF2 target gene expression via the regulation of HDAC5 phosphorylation at Ser259/498, and suggest that HDAC5 could be a potential mechanism contributing to the therapeutic actions of EPO and cEPO.

60 APPLIED LIFE SCIENCES↗

Simulation of anoxic lenses as exporters of reactivity in alluvial aquifer sediments

Sedimentary interfaces between contrasting hydrogeological facies in alluvial aquifers drive the development of biogeochemical interfaces that influence subsurface and surface water quality. Here, we calibrate a reactive transport model on a series of dual-domain column experiments, where centimeter-scale, low-permeability, organic-rich anoxic lenses are embedded in coarser-grained aquifer material. Simulations explicitly account for C, Fe, and S cycling at the interface between the lenses and the aquifer in water-saturated conditions. Our results highlight the role of fine-grained, organic-rich inclusions not only as sources or sinks for redox-sensitive species, but also as exporters of nutrients that stimulate downgradient biogeochemical cycling. By releasing large amounts of organic carbon into the surrounding aquifer, such lenses drive the development of proximal secondary reduction zones ("halos"), characterized by high microbial activity (e.g., sulfate reduction) and accumulation of reduced reaction products (e.g., iron sulfide). If similar secondary reduction zones develop within the hydraulically conductive domain of an aquifer, they would be highly susceptible to changes in hydrologic conditions, for instance oxygen pulses associated with seasonal snowmelt. Furthermore, our results also emphasize the limitations of relying solely on aqueous species measurements to inform reactivity in systems where fast redox cycling and/or sizeable particulate transport may limit the signature of reactivity in the dissolved phase.

54 ENVIRONMENTAL SCIENCES↗

HIV-1 pre-mRNA commitment to Rev mediated export through PSF and Matrin 3

Human immunodeficiency virus gene expression and replication are regulated at several levels. Incompletely spliced viral RNAs and full-length genomic RNA contain the RRE element and are bound by the viral trans-acting protein Rev to be transported out of the nucleus. Previously we found that the nuclear matrix protein MATR3 was a cofactor of Rev-mediated RNA export. Here we show that the pleiotropic protein PSF binds viral RNA and is associated with MATR3. PSF is involved in the maintenance of a pool of RNA available for Rev activity. However, while Rev and PSF bind the viral pre-mRNA at the site of viral transcription, MATR3 interacts at a subsequent step. We propose that PSF and MATR3 define a novel pathway for RRE-containing HIV-1 RNAs that is hijacked by the viral Rev protein.

60 APPLIED LIFE SCIENCES↗

hnRNP A2/B1 interacts with influenza A viral protein NS1 and inhibits virus replication potentially through suppressing NS1 RNA/protein levels and NS1 mRNA nuclear export

The NS1 protein of influenza viruses is a major virulence factor and exerts its function through interacting with viral/cellular RNAs and proteins. In this study, we identified heterogeneous nuclear ribonucleoprotein A2/B1 (hnRNP A2/B1) as an interacting partner of NS1 proteins by a proteomic method. Knockdown of hnRNP A2/B1 by small interfering RNA (siRNA) resulted in higher levels of NS vRNA, NS1 mRNA, and NS1 protein in the virus-infected cells. In addition, we demonstrated that hnRNP A2/B1 proteins are associated with NS1 and NS2 mRNAs and that knockdown of hnRNP A2/B1 promotes transport of NS1 mRNA from the nucleus to the cytoplasm in the infected cells. Lastly, we showed that knockdown of hnRNP A2/B1 leads to enhanced virus replication. Our results suggest that hnRNP A2/B1 plays an inhibitory role in the replication of influenza A virus in host cells potentially through suppressing NS1 RNA/protein levels and NS1 mRNA nucleocytoplasmic translocation. - Highlights: • Cellular protein hnRNP A2/B1 interacts with influenza viral protein NS1. • hnRNP A2/B1 suppresses the levels of NS1 protein, vRNA and mRNA in infected cells. • hnRNP A2/B1 protein is associated with NS1 and NS2 mRNAs. • hnRNP A2/B1 inhibits the nuclear export of NS1 mRNAs. • hnRNP A2/B1 inhibits influenza virus replication.

60 APPLIED LIFE SCIENCES↗

Knowledge-guided graph machine learning for spatially distributed prediction of daily discharge and nitrogen export dynamics

Spatially distributed prediction of streamflow and nitrogen export dynamics is essential for precision management of agricultural watersheds. While temporal deep learning models such as Long Short-Term Memory (LSTM) have shown strong performance at basin scales, their ability to generalize spatially is limited by insufficient representation of spatial dependencies and flow paths, particularly under data-scarce conditions. To address this gap, we propose HydroGraphNet, a knowledge-guided graph machine learning framework that integrates process-based knowledge and explicit spatial learning into temporal modeling. This framework incorporates directed graph topology to encode watershed connectivity and upstream inflows, with mass balance constraints to improve physical consistency. To enhance generalization in sparsely monitored regions, HydroGraphNet is pretrained on synthetic data generated by the SWAT+ (Soil and Water Assessment Tool Plus) model. We evaluated HydroGraphNet in the Upper Sangamon River Basin (44 HUC-12 subwatersheds, 2001–2020) against two LSTM baselines: a lumped basin-level model and a distributed variant. When benchmarked on SWAT+ simulations in pretraining, HydroGraphNet improved test NSEs by 8.9% (discharge) and 13.7% (NO₃–N load) in temporal extrapolation, and by 27.1% and 34.7% in spatial extrapolation, relative to the Lumped LSTM baseline. After fine-tuning with USGS monitoring data, the model achieved mean test NSE (KGE) scores of 0.768 (0.861) for discharge and 0.626 (0.664) for NO₃–N load, substantially outperforming baselines. Attribution analysis further highlighted the importance of upstream inflow representation and graph-based spatial learning in capturing cross-subwatershed dependencies. The model also reproduced seasonal hydrological and biogeochemical patterns consistent with known processes, demonstrating its robustness and process fidelity for spatially distributed prediction. Altogether, HydroGraphNet advances the integration of physical knowledge and spatially explicit learning in hydrological modeling, offering a generalizable framework for distributed modeling to support spatially targeted water quality management in data-scarce watersheds.

54 ENVIRONMENTAL SCIENCES↗

Machine Learning Models to Predict Inhibition of the Bile Salt Export Pump

Cholestatic liver injury is frequently associated with drug inhibition of bile salt transporters, such as the bile salt export pump (BSEP). Reliable in silico models to predict BSEP inhibition directly from chemical structures would significantly reduce costs during drug discovery and could help avoid injury to patients. We report our development of classification and regression models for BSEP inhibition with substantially improved performance over previously published models. We assessed the performance effects of different methods of chemical featurization, data set partitioning, and class labeling and identified the methods producing models that generalized best to novel chemical entities.

59 BASIC BIOLOGICAL SCIENCES↗

Geochemical Exports to River From the Intrameander Hyporheic Zone Under Transient Hydrologic Conditions: East River Mountainous Watershed, Colorado

To understand how redox processes influence carbon, nitrogen, and iron cycling within the intrameander hyporheic zone, we developed a biotic and abiotic reaction network and incorporated it into the reactive transport simulator PFLOTRAN. Additionally, two-dimensional reactive flow and transport simulations were performed (1) to evaluate how transient hydrological conditions control the lateral redox zonation within an intrameander region of the East River in Colorado and (2) to quantify the impact of a single meander on subsurface exports of carbon and other geochemical species to the river. The meander's overall contribution to the river was quantified by integrating geochemical outfluxes along the outside of the meander bend. The model was able to capture the field-observed trends of dissolved oxygen, nitrate, iron, pH, and total inorganic carbon along a 2-D transect. Consistent with field observations, simulated dissolved oxygen and nitrate decreased along the intrameander flow paths while iron (Fe 2+ ) concentration increased. The simulation results further demonstrated that the reductive potential of the lateral redox zonation was controlled by groundwater velocities resulting from river stage fluctuations, with low-water conditions promoting reducing conditions. The sensitivity analysis results showed that permeability had a more significant impact on biogeochemical zonation compared to the reaction pathways under transient hydrologic conditions. Lastly, the simulation results further indicated that the meander acted as a sink for organic and inorganic carbon as well as iron during the extended baseflow and high-water conditions; however, these geochemical species were released into the river during the falling limb of the hydrograph.

58 GEOSCIENCES↗

High Frequency Monitoring and Nitrate Sourcing Reveals Baseflow and Stormflow Controls on Total Dissolved Nitrogen and Carbon Export Along a Rural‐Urban Gradient

Abstract Efforts to reduce nitrogen and carbon loading from developed watersheds typically target specific flows or sources, but across gradients in development intensity there is no consensus on the contribution of different flows to total loading or sources of nitrogen export. This information is vital to optimize management strategies leveraging source reductions, stormwater controls, and restorations. We investigate how solute loading and sources vary across flows and land‐use using high frequency monitoring and stable nitrate isotope analysis from five catchments with different sanitary infrastructure, along a gradient in development intensity. High frequency monitoring allowed estimation of annual loading and attribution to storm versus baseflows. Nitrate loads were 16 kg/km 2 /yr. from the forested catchment and ranged from 68 to 119 kg/km 2 /yr., across developed catchments, highest for the septic served site. Across developed catchments, baseflow contributions ranged from 40% of N loading to 75% from the septic served catchment, and the contribution from high stormflows increased with development intensity. Stormflows mobilized and mixed many surface and subsurface nitrate sources while baseflow nitrate was dominated by fewer sources which varied by catchment (soil, wastewater, or fertilizer). To help inform future sampling designs, we demonstrate that grab sampling and targeted storm sampling would likely fail to accurately predict annual loadings within the study period. The dominant baseflow loads and subsurface stormflows are not treated by surface water management practices primarily targeted to surface stormflows. Using a balance of green and gray infrastructure and stream/riparian restoration may target specific flow paths and improve management.

Delesantro, Joseph M.↗

When Do Riverine Systems “Feel the Burn”? Simulating How Burn Extent and Severity Modulate Hydrologic Controls on Biogeochemical Export

Wildfires impact terrestrial landscapes and downstream river corridors through shifts in vegetation and soil properties leading to downstream hydrologic and water quality impacts. The magnitude of these impacts depend on a complex and interconnected set of wildfire, landscape, and aquatic processes. Here, we isolate the impact of post-fire hydrologic changes on streamflow, nitrate, and dissolved organic carbon using the Soil and Water Assessment Tool (SWAT) model. We explore how responses differ across burn severity and area burned in two test basins: a humid forested basin and a semi-arid mixed land use basin. We ran 1830 wildfire simulations testing impacts of area burned, burn severity, and post-fire precipitation on streamflow, nitrate, and dissolved organic carbon. Our work suggests that area burned thresholds differ with burn severity and analyte. Additionally, post-fire transport of dissolved organic carbon was sensitive to both area burned and severity, while nitrate was primarily sensitive to area burned. Despite a muted (−9.5 to 5.7 mm yr −1 change) hydrologic response in the semi-arid basin, the model predicted large (7%–288% increase) shifts in dissolved organic carbon, suggesting that post-fire shifts in flow pathways and soil properties are key in its response. The limited shifts in nitrate responses in the simulations highlight that terrestrial post-fire transformations, rather than hydrologic changes, may control the increases in stream nitrate often observed post-fire. As wildfire regimes are shifting, improving understanding of post-fire nutrient export responses is critical to protect freshwater resources and aquatic ecosystems.

Wampler, Katherine A. [Pacific Northwest National ↗

Mechanistic determinants of the directionality and energetics of active export by a heterodimeric ABC transporter

The ATP-binding cassette (ABC) transporter associated with antigen processing (TAP) participates in immune surveillance by moving proteasomal products into the endoplasmic reticulum (ER) lumen for major histocompatibility complex class I loading and cell surface presentation to cytotoxic T cells. Here we delineate the mechanistic basis for antigen translocation. Notably, TAP works as a molecular diode, translocating peptide substrates against the gradient in a strict unidirectional way. We reveal the importance of the D-loop at the dimer interface of the two nucleotide-binding domains (NBDs) in coupling substrate translocation with ATP hydrolysis and defining transport vectoriality. Substitution of the converved aspartate, which coordinates the ATP-binding site, decreases NBD dimerization affinity and turns the unidirectional primary active pump into a passive bidirectional nucleotide-gated facilitator. Thus, ATP hydrolysis is not required for translocation per se , but is essential for both active and unidirectional transport. As a result, our data provide detailed mechanistic insight into how heterodimeric ABC exporters operate.

59 BASIC BIOLOGICAL SCIENCES↗

Recognition of nuclear export signals by CRM1 carrying the oncogenic E571K mutation

The E571K mutation of CRM1 is highly prevalent in some cancers, but its mechanism of tumorigenesis is unclear. Glu571 of CRM1 is located in its nuclear export signal (NES)-binding groove, suggesting that binding of select NESs may be altered. We generated HEK 293 cells with either monoallelic CRM1WT/E571K or biallelic CRM1E571K/E571K using CRISPR/Cas9. We also combined analysis of binding affinities and structures of 27 diverse NESs for wild-type and E571K CRM1 with structure-based bioinformatics. While most NESs bind the two CRM1 similarly, NESs from Mek1, eIF4E-transporter, and RPS2 showed >10-fold affinity differences. These NESs have multiple charged side chains binding close to CRM1 position 571, but this feature alone was not sufficient to predict different binding to CRM1(E571K). Consistent with eIF4E-transporter NES binding weaker to CRM1(E571K), eIF4E-transporter was mislocalized in tumor cells carrying CRM1(E571K). This serves as proof of concept that understanding how CRM1(E571K) affects NES binding provides a platform for identifying cargoes that are mislocalized in cancer upon CRM1 mutation. Finally, we showed that large affinity changes seen with some NES peptides (of Mek1 and RPS2) do not always translate to the full-length cargoes, suggesting limitations with current NES prediction methods. Therefore, comprehensive studies like ours are imperative to identify CRM1 cargoes with real pathogenic potential.

59 BASIC BIOLOGICAL SCIENCES↗