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At least 289 records · Page 16

Enhanced spontaneous DNA twisting/bending fluctuations unveiled by fluorescence lifetime distributions promote mismatch recognition by the Rad4 nucleotide excision repair complex

Rad4/XPC recognizes diverse DNA lesions including ultraviolet-photolesions and carcinogen-DNA adducts, initiating nucleotide excision repair. Studies have suggested that Rad4/XPC senses lesion-induced helix-destabilization to flip out nucleotides from damaged DNA sites. However, characterizing how DNA deformability and/or distortions impact recognition has been challenging. Here, using fluorescence lifetime measurements empowered by a maximum entropy algorithm, we mapped the conformational heterogeneities of artificially destabilized mismatched DNA substrates of varying Rad4-binding specificities. The conformational distributions, as probed by FRET between a cytosine-analog pair exquisitely sensitive to DNA twisting/bending, reveal a direct connection between intrinsic DNA deformability and Rad4 recognition. High-specificity CCC/CCC mismatch, free in solution, sampled a strikingly broad range of conformations from B-DNA-like to highly distorted conformations that resembled those observed with Rad4 bound; the extent of these distortions increased with bound Rad4 and with temperature. Conversely, the non-specific TAT/TAT mismatch had a homogeneous, B-DNA-like conformation. Molecular dynamics simulations also revealed a wide distribution of conformations for CCC/CCC, complementing experimental findings. We propose that intrinsic deformability promotes Rad4 damage recognition, perhaps by stalling a diffusing protein and/or facilitating ‘conformational capture’ of pre-distorted damaged sites. Surprisingly, even mismatched DNA specifically bound to Rad4 remains highly dynamic, a feature that may reflect the versatility of Rad4/XPC to recognize many structurally dissimilar lesions

59 BASIC BIOLOGICAL SCIENCES↗

MECHANISMS OF RADIATION CARCINOGENESIS: WHAT IS REALLY INDUCED?

Abstract It has been difficult to understand why the relative risk for cancer decreases with an increase in time since an exposure to radiation. It was recently recognized that this decline can be explained by a parallel shift of the age-related cancer mortality curve toward younger ages. In fact, it has been known for many years that mouse survival curves exhibit a parallel shift toward younger ages following an exposure to radiation, but it was not recognized that the mutation induction theory is incompatible with this parallel shift. This is because a parallel shift in the survival curve implies that all the irradiated individuals are affected, but the mutation induction theory assumes that only a fraction of the irradiated individuals is affected following an exposure to radiation. Thus, it seems likely that the target of radiation action, which leads to carcinogenesis, is not restricted to epithelial cells but includes all of the surrounding cells leading to an altered microenvironment. Since it is repeatedly observed that radiation-exposed normal tissues can stimulate transplanted or spontaneously arising tumor cells to grow faster, worsen the malignant phenotypes and finally kill the host earlier than usual, an exposure to radiation seems most likely to cause tissue inflammation, which creates conditions favorable for the growth of spontaneously arising tumor cells. This new concept suggests that it might be possible to attenuate the extent of radiation carcinogenesis by intervening in tissue inflammatory processes.

38 RADIATION CHEMISTRY, RADIOCHEMISTRY, AND NUCLEA↗

SNAPPy: A snakemake pipeline for scalable HIV-1 subtyping by phylogenetic pairing

Abstract Human immunodeficiency virus 1 (HIV-1) genome sequencing is routinely done for drug resistance monitoring in hospitals worldwide. Subtyping these extensive datasets of HIV-1 sequences is a critical first step in molecular epidemiology and evolution studies. The clinical relevance of HIV-1 subtypes is increasingly recognized. Several studies suggest subtype-related differences in disease progression, transmission route efficiency, immune evasion, and even therapeutic outcomes. HIV-1 subtyping is mainly done using web-servers. These tools have limitations in scalability and potential noncompliance with data protection legislation. Thus, the aim of this work was to develop an efficient method for large-scale local HIV-1 subtyping. We designed SNAPPy: a snakemake pipeline for scalable HIV-1 subtyping by phylogenetic pairing. It contains several tasks of phylogenetic inference and BLAST queries, which can be executed sequentially or in parallel, taking advantage of multiple-core processing units. Although it was built for subtyping, SNAPPy is also useful to perform extensive HIV-1 alignments. This tool facilitates large-scale sequence-based HIV-1 research by providing a local, resource efficient and scalable alternative for HIV-1 subtyping. It is capable of analyzing full-length genomes or partial HIV-1 genomic regions (GAG, POL, and ENV) and recognizes more than ninety circulating recombinant forms. SNAPPy is freely available at: https://github.com/PMMAraujo/snappy/releases.

Araújo, Pedro M. M.↗

2022 Prentice Award Lecture: Advancing Retinal Imaging and Visual Function in Patient Management and Disease Mechanisms

Patient-based research plays a key role in probing basic visual mechanisms. Less-well recognized is the role of patient-based retinal imaging and visual function studies in elucidating disease mechanisms, which are accelerated by advances in imaging and function techniques and are most powerful when combined with the results from histology and animal models. A patient's visual complaints can be one key to patient management, but human data are also key to understanding disease mechanisms. Unfortunately, pathological changes can be difficult to detect. Before advanced retinal imaging, the measurement of visual function indicated the presence of pathological changes that were undetectable with existing clinical examination. Over the past few decades, advances in retinal imaging have increasingly revealed the unseen. This has led to great strides in the management of many diseases, particularly diabetic retinopathy and macular edema, and age-related macular degeneration. It is likely widely accepted that patient-based research, as in clinical trials, led to such positive outcomes. Both visual function measures and advanced retinal imaging have clearly demonstrated differences among retinal diseases. Contrary to initial thinking, sight-threatening damage in diabetes occurs to the outer retina and not only to the inner retina. This has been clearly indicated in patient results but has only gradually entered the clinical classifications and understanding of disease etiology. There is strikingly different pathophysiology for age-related macular degeneration compared with photoreceptor and retinal pigment epithelial genetic defects, yet research models and even some treatments confuse these. It is important to recognize the role that patient-based research plays in probing basic visual mechanisms and elucidating disease mechanisms, combining these findings with the concepts from histology and animal models. Thus, this article combines sample instrumentation from my laboratory and progress in the fields of retinal imaging and visual function.

60 APPLIED LIFE SCIENCES↗

MBP-binding DARPins facilitate the crystallization of an MBP fusion protein

The production of high-quality crystals is the main bottleneck in determining the structures of proteins using X-ray crystallography. In addition to being recognized as a very effective solubility-enhancing fusion partner, Escherichia coli maltose-binding protein (MBP) has also been successfully employed as a `fixed-arm' crystallization chaperone in more than 100 cases. Here, it is reported that designed ankyrin-repeat proteins (DARPins) that bind with high affinity to MBP can promote the crystallization of an MBP fusion protein when the fusion protein alone fails to produce diffraction-quality crystals. As a proof of principle, three different co-crystal structures of MBP fused to the catalytic domain of human dual-specificity phosphatase 1 in complex with DARPins are reported.

Gumpena, Rajesh↗

Underwater Fish Detection using Deep Learning for Water Power Applications

Clean energy from oceans and rivers is becoming a reality with the development of new technologies like tidal and instream turbines that generate electricity from naturally flowing water. These new technologies are being monitored for effects on fish and other wildlife using underwater video. Methods for automated analysis of underwater video are needed to lower the costs of analysis and improve accuracy. We have trained a deep learning model, YOLO, to recognize fish in underwater video using three very different datasets recorded at real-world water power sites. Training and testing with examples from all three datasets resulted in a mean average precision (MAP) score of 0.5392. To test how well a model could generalize to new datasets, the model was trained using examples from only two of the datasets and then tested on examples from all three datasets. The resulting model could not recognize fish in the dataset that was not part of the training set. The MAP scores on the other two datasets that were included in the training set were higher than the scores achieved by the model trained on all three datasets. These results indicate that different methods are needed in order to produce a trained model that can generalize to new data sets such as those encountered in real world applications.

fish detection, underwater video, marine and hydro↗

Frequency Stability Enhancement in a Fully Solar Power Grid: A Case Study on the Saudi Model

Recently, renewable energy sources (RES) such as solar photovoltaic (PV) have been recognized for their significant potential. Several studies have explored the issue of frequency stability with high RES penetration. A significant observation related to this issue is the decline in the weighted short circuit ratio (WSCR), which has the potential to adversely affect system stability. This research uniquely focuses on the gradual transition of the Saudi Electricity Company (SEC) system to a fully integrated solar PV grid, offering insight and methodology that can be adapted for other power systems. While a variety of mitigation methods are available, synchronous condensers (SC) were used in this study to stabilize grid frequency, and their implementation has been shown to improve the overall performance of the system. In response to the observed decline in WSCR, a comprehensive short circuit analysis was applied across all regions in the Saudi power grid model to evaluate the reduction in short circuit power and highlight areas requiring enhancement. Finally, scenarios illustrating frequency response with a gradual increase in SCR were analyzed, with the objective of establishing a stable, fully solar PV power system that is resilient to sudden power grid disturbances.

14 SOLAR ENERGY↗

On Distribution Grid Optimal Power Flow Development and Integration

Due to changes in electric distribution grid operation, new operation regimes have been recommended. Distribution grid optimal power flow (DOPF) has received tremendous attention in the research community, yet it has not been fully adopted across the utility industry. Our paper recognizes this problem and suggests a development and integration procedure for DOPF. We propose development of DOPF as a three step procedure of 1) processing the grid, 2) obtaining a tractable solution, and 3) implementing multiple solution algorithms and benchmarking them to improve application reliability. For the integration of DOPF, we demonstrate how a DOPF federate may be developed that can be integrated in a co-simulation environment to mimic the real-world conditions and hence improve its practicality to be deployed in the field. To demonstrate the efficacy of the proposed methods, tests on IEEE 123 bus system are performed where the usage of tractable formulation in DOPF algorithm development and its comparison to the benchmark solution are demonstrated.

Hanif, Sarmad↗

Recognition of dominant attractants by key chemoreceptors mediates recruitment of plant growth‐promoting rhizobacteria

Summary Chemotaxis to plant root exudates is supposed to be a prerequisite for efficient root colonization by rhizobacteria. This is a highly multifactorial process since root exudates are complex compound mixtures of which components are recognized by different chemoreceptors. Little information is available as to the key components in root exudates and their receptors that drive colonization related chemotaxis. We present here the first global assessment of this issue using the plant growth‐promoting rhizobacterium (PGPR) Bacillus velezensis SQR9 (formerly B. amyloliquefaciens ). This strain efficiently colonizes cucumber roots, and here, we show that chemotaxis to cucumber root exudates was essential in this process. We conducted chemotaxis assays using cucumber root exudates at different concentrations, individual exudate components as well as recomposed exudates, taking into account their concentrations detected in root exudates. Results indicated that two key chemoreceptors, McpA and McpC, were essential for root exudate chemotaxis and root colonization. Both receptors possess a broad ligand range and recognize most of the exudate key components identified (malic, fumaric, gluconic and glyceric acids, Lys, Ser, Ala and mannose). The remaining six chemoreceptors did not contribute to exudate chemotaxis. This study provides novel insight into the evolution of the chemotaxis system in rhizobacteria.

Feng, Haichao↗

Rapid Prediction of Low (<1%) trans Fat Content in Edible Oils and Fast Food Lipid Extracts by Infrared Spectroscopy and Partial Least Squares Regression

Abstract The United States Food and Drug Administration (FDA) ruled that partially hydrogenated oils (PHO), the major dietary source of industrially produced trans fat (TF), were no longer “generally recognized as safe (GRAS)” for any use in human food. Consequently, the objective of this study was to develop a rapid screening procedure using attenuated total reflection Fourier‐transform infrared (ATR‐FTIR) spectroscopy in conjunction with partial least squares regression (PLSR) for the quantitative and accurate prediction of low concentrations of trans fatty acid (TFAs) (<1% of total fatty acids (FAs)). Broad‐based calibration models were developed for a combined set of samples consisting of edible oils and fast food lipid extracts. Predicted concentrations of TFAs in the two matrices showed good correlation with the primary reference data generated by gas chromatography (GC) ( R 2 > 0.99) and high accuracy as evidenced by low root‐mean‐square error of cross‐validation (RMSECV) values. The lowest TFA concentration, determined by GC to be 0.13% of total FAs, was accurately predicted by ATR‐FTIR/PLSR as 0.18% of total FAs. This simple, rapid ATR‐FTIR/PLSR methodology has the potential for use as a screening alternative to conventional gas chromatographic methods for predicting the TFA content of edible oils and food lipid extracts for regulatory purposes and quality control of raw material and processed food. Practical Applications F DA ruled that partially hydrogenated oils were no longer “generally recognized as safe (GRAS)” for any use in human food. Consequently, we have proposed a rapid screening procedure, based on infrared spectroscopy and chemometrics, to rapidly and accurately predict low concentrations of trans fatty acids (<1% of total fatty acids) in edible oils and food lipid extracts.

Karunathilaka, Sanjeewa R.↗

Climate reconstruction for the Entre‐Douro‐e‐Minho region ( NW Portugal) between AD 1626 and AD 1820: synthesis of viticulture data and foraminiferal evidence

This work presents two novel climate‐related time series for the northwest of Portugal. The first is an AD 1626–1820 triennial‐resolved wine production series, based on the Benedictine accounts from six monasteries of the Entre‐Douro‐e‐Minho ( EDM ) region. The second, an AD 1654–2010 benthic foraminiferal record from the Caminha salt marsh, located in the lower estuary of the Minho River. The series were analysed together for the common period to outline how both palaeoclimatic proxies respond to the most likely natural environmental drivers of temporal variability, solar forcing included. Singular spectral analysis revealed a common significant multidecadal periodicity agreeing with recognized long‐term changes in solar activity, i.e. the Lower Gleissberg cycle (50–80 years). The application of wavelet analysis allowed the detection of high coherence at this time scale (centred at c . 64 years) between marsh foraminifera and both total solar irradiance and the North Atlantic Oscillation index. This relationship persists throughout the c . AD 1730–1875 period. The continuous wavelet transform results for wine production were inconclusive. As the time‐span analysed is recognized as one of high socio‐economic and political distress, the main human‐driven impacts on wine production, particularly in the two periods of greatly reduced solar activity – the Maunder and Dalton Minima – are reviewed in the light of the available historical records. In addition to a documented climate‐related agricultural crisis in Portugal, damage and losses to wine production may have been triggered by several local and international conflicts in which the country was involved. But to what extent the two influences contributed to the wine production variations observed in the EDM region during both periods remains an open question.

Moreno, João↗

Sedimentation of ballasted cells-free EPS in meromictic Fayetteville Green Lake

Fayetteville Green Lake (FGL) is a recognized, extensively studied present-day model of the stratified Proterozoic Ocean. Nonetheless, biomass sedimentation in FGL remains hard to explain: Whilst virtually all sediment pigments belong to photosynthetic sulfur bacteria (PSB) from a chemocline, the isotopic carbon signature of the bulk biomass suggests its epilimnetic phytoplankton origin. Here we present experimental evidence that FGL Synechococci produce copious extracellular polysacchrides (EPS), which become impregnated with Ca, Mg and Na cations and are release to the environment as ballasted cell coverings. We propose that sedimentation of these cell-free EPS can constitute the bulk of pigment-less organic material in FGL sediment. Because increased availability of inorganic carbon (Ci) stimulates accumulation of ballasted EPS, we propose the universal role of cyanobacterial EPS in biomass sedimentation in the high-Ci Paleoproterozoic Ocean as well as in modern aquatic systems like FGL.

Kamennaya, Nina A.↗

Powdery mildew effectors AVR A1 and BEC1016 target the ER J‐domain protein Hv ERdj3B required for immunity in barley

Abstract The barley powdery mildew fungus, Blumeria hordei (Bh), secretes hundreds of candidate secreted effector proteins (CSEPs) to facilitate pathogen infection and colonization. One of these, CSEP0008, is directly recognized by the barley nucleotide‐binding leucine‐rich‐repeat (NLR) receptor MLA1 and therefore is designated AVR A1 . Here, we show that AVR A1 and the sequence‐unrelated Bh effector BEC1016 (CSEP0491) suppress immunity in barley. We used yeast two‐hybrid next‐generation interaction screens (Y2H‐NGIS), followed by binary Y2H and in planta protein–protein interactions studies, and identified a common barley target of AVR A1 and BEC1016, the endoplasmic reticulum (ER)‐localized J‐domain protein Hv ERdj3B. Silencing of this ER quality control (ERQC) protein increased Bh penetration. Hv ERdj3B is ER luminal, and we showed using split GFP that AVR A1 and BEC1016 translocate into the ER signal peptide‐independently. Overexpression of the two effectors impeded trafficking of a vacuolar marker through the ER; silencing of Hv ERdj3B also exhibited this same cellular phenotype, coinciding with the effectors targeting this ERQC component. Together, these results suggest that the barley innate immunity, preventing Bh entry into epidermal cells, requires ERQC. Here, the J‐domain protein Hv ERdj3B appears to be essential and can be regulated by AVR A1 and BEC1016. Plant disease resistance often occurs upon direct or indirect recognition of pathogen effectors by host NLR receptors. Previous work has shown that AVR A1 is directly recognized in the cytosol by the immune receptor MLA1. We speculate that the AVR A1 J‐domain target being inside the ER, where it is inapproachable by NLRs, has forced the plant to evolve this challenging direct recognition.

54 ENVIRONMENTAL SCIENCES↗

From glomalin to glomalose: unraveling the molecular identity of the MAb32B11 antigen

Glomalin, a substance produced by arbuscular mycorrhizal (AM) fungi, haswell-documented benefits for plant and soil health, including water retention and soil aggre-gation. Glomalin quantification has been performed by enzyme-linked immunosorbent assay(ELISA) using a monoclonal antibody, MAb32B11, that has been described as targeting a heatshock protein 60 (RiHSP60). In this study, we re-examined the molecular nature of the antigen recognized byMAb32B11. MAb32B11 did not cross-react with the RiHSP60 polypeptide. Glomalin extracts of Rhizo-phagus irregularis showed strong and dose-dependent cross-reactivity with MAb32B11 evenwhen protein levels were undetectable, raising doubts about the proteinaceous nature of theantigen. Protease treatments of glomalin extracts did not affect the ELISA signal. However,treatment with periodate, which degrades polysaccharides, significantly reduced the signal. Astrong correlation between carbohydrate content and the ELISA signal was observed in glo-malin extracts. These findings indicate that MAb32B11 recognizes a carbohydrate, likely originating fromcell walls of AM fungi. Further analysis of glomalin extracts using size exclusion chromatogra-phy suggests that the epitope of MAb32B11 is a complex carbohydrate in the size range of511–600 kDa. Understanding the true nature of glomalin will enhance our ability to quantifyit accurately and leverage its agricultural benefits.

59 BASIC BIOLOGICAL SCIENCES↗

Reman Co-Design: A Combined Design and Remanufacturing Optimization Framework for the Sustainable Design of High-Value Components

Remanufacturing is a process that returns end-of-life equipment to as-new conditions and offers numerous environmental and economic benefits. To fully capitalize on remanufacturing, its synergistic interactions with design must be fully realized and addressed during the design stage. Although this fact is widely recognized in the literature, most of the current studies focus primarily either on the design or remanufacturing aspects of design for remanufacturing (DfRem). In an effort to offer a more integrated DfRem approach than those reported in the literature, we propose a new combined design and remanufacturing optimization (reman co-design) framework that takes a holistic approach by leveraging the intricate interplay between design and remanufacturing. Here, the aim of this formulation is to identify the optimal decisions that maximize the benefits of remanufacturing throughout the entire lifespan of a product. To showcase the utility of the new formulation, we are using a case study of a hydraulic manifold, (re)manufactured by John Deere. Using this industry example, we compare the results of reman co-design to the ones from a decoupled remanufacturing design approach. Results reveal that remanufacturing benefits are better realized and improved upon when using the developed reman co-design approach.

design for X↗

Integrated Sustainable Product Design With Warranty and End-of-Use Considerations

The concept of integrated sustainable product design has recently emerged, aiming to incorporate downstream life cycle performance into the initial product design to enhance sustainability. Various sustainable product design tools based on life cycle assessment or quality function deployment have been established while the impact of reliability on circular practices has received limited attention. Recognizing the critical role of product reliability in post-design performance, this paper develops a product design optimization model that considers the warranty performance and the effect of end-of-use options. The model takes into account the effect of uncertain operating conditions on product reliability. Two optimization goals including the minimization of expected unit life cycle cost and environmental impact are achieved by the model. To demonstrate the benefits of the integrated approach, the model is applied to an electric motor design problem. The results highlight that integrating end-of-use options in the early design phase leads to adjustments in component selection and reliability design. Moreover, the circular utilization of used products enables cost savings throughout the product’s life cycle and contributes to environmental impact reduction. Finally, the study analyzes the effects of operating conditions, warranty policies, and take-back prices for used products on design decisions, providing valuable insights for product designers.

Integrated design↗

Toward unrestricted use of public genomic data

Despite some notable progress in data sharing policies and practices, restrictions are still often placed on the open and unconditional use of various genomic data after they have received official approval for release to the public domain or to public databases. These restrictions, which often conflict with the terms and conditions of the funding bodies who supported the release of those data for the benefit of the scientific community and society, are perpetuated by the lack of clear guiding rules for data usage. Existing guidelines for data released to the public domain recognize but fail to resolve tensions between the importance of free and unconditional use of these data and the “right” of the data producers to the first publication. This self-contradiction has resulted in a loophole that allows different interpretations and a continuous debate between data producers and data users on the use of public data. We argue that the publicly available data should be treated as open data, a shared resource with unrestricted use for analysis, interpretation, and publication.

59 BASIC BIOLOGICAL SCIENCES↗