Paediatric palliative care (PPC) in oncology plays a crucial role in enhancing the quality of life for children with cancer and their families. However, outcome measurement in this context remains fragmented, with tools often lacking validation, paediatric specificity, or contextual relevance, especially in low- and middle-income countries (LMICs). This scoping review maps the outcome measurement tools used in paediatric oncology palliative care, examining their assessed domains, psychometric properties and cultural, linguistic and health-system contextual relevance, particularly in LMICs. We systematically searched six databases from 2006 to 2025 and identified 27 eligible studies reporting on 28 unique tools across six key domains: symptom burden, quality of life, psychological well-being, spiritual health, caregiver burden and end-of-life care. While tools like Paediatric Quality of Life Inventory (PedsQL) and Symptom Screening in Pediatrics Tool (SSPedi) showed strong psychometric performance, only a few had been adapted for LMIC contexts. The psychological and spiritual domains were underrepresented, and most tools relied on proxy reporting, which limited child-centred assessments. The findings suggest the need for validated, culturally sensitive and inclusive tools co-developed with children and caregivers. Establishing such outcome measures is essential to ensuring equitable, high-quality palliative care across diverse settings. This review lays the groundwork for developing tools and advancing policies that support comprehensive and compassionate care for children with cancer. Review registration: The review is registered on Open Science Framework (OSF) https://doi.org/10.17605/OSF.IO/G8BN3 .
The prediction of a biological sequence's function is an exploding field that could prove to be an invaluable tool in deciphering molecular mechanisms of diseases and genetic variation. However, existing techniques suffer from a very low number of experimentally verified labels as well as the complexity of modeling sequence context. Previous work only focuses on semi-supervised learning (SSL) or pre-trained language models (PLMs) individually and does not consider their complementarity: whereas PLMs capture syntactic conservatism and long-range dependency from huge unlabeled sequence data as features, SSL updates label-level certainty via confidence-weighted pseudo-label selection. To realize integrating these benefits, we make a creative use of PLMs as strong feature extractors to make the sequence semantics extracted, and meanwhile leverage the SSL to constrain the decision boundary by selecting reliable pseudo-labels. We show that our framework achieves competitive performance compared to full supervision while using far fewer labeled samples, on two biological prediction tasks: DNA-binding protein (DBP) and non-coding RNA (ncRNA) detection. In general, our approach, developed here, not only offers an efficient solution for discovering new DBP and ncRNA in low-resource settings where experimental verification is infeasible, but also lays a solid methodological foundation for biological sequence classification in computational biology. Key points Unlabeled data significantly improves model accuracy in semi-supervised learning. The language model-based self-training method outperforms traditional semi-supervised approaches like TSVM. It achieves competitive performance against state-of-the-art fully supervised methods, requiring only 1% labeled data for effective classification and accurately identifying novel biomolecules.
The white-spotted flower chafer (Protaetia brevitarsis) exhibits a preference for feeding on mature fruits of various crops, making it a significant agricultural pest. Pesticide application often results in fruit residues and environmental contamination, highlighting the need for eco-friendly and highly effective alternative control strategies. For instance, targeted regulation of the pest's olfactory system could be used to influence its behavior. This study focused on the enzymes responsible for terminating olfactory signals, namely odorant degrading enzymes (ODEs). Through antennal transcriptome sequencing, four related gene families comprising a total of 94 genes were identified, including 23 carboxylesterases, 40 cytochrome P450s, 21 glutathione S-transferases, and 10 UDP-glycosyltransferases. The majority of enzyme proteins possessed typical domain characteristics and key residues, suggesting their potential catalytic activity. These enzymes clustered into multiple clades in the phylogenetic tree, likely undertaking diverse physiological functions including odor substance metabolism. To more precisely determine the ODE candidates, tissue expression profiling was conducted on 17 highly expressed genes (FPKM >100). The results revealed that 16 genes were enriched or specifically expressed in the antennae. We propose these genes as primary candidates for ODEs, serving as a prioritized list for future functional validation. Our work lays the foundation for elucidating the odor signal inactivation mechanism in P. brevitarsis, expands the transcriptomic resource of ODEs in Scarabaeidae, and provides a theoretical basis for developing novel pest management targets based on olfactory behavior.
AbstractMale-produced seminal fluid proteins modify female behavior and physiology. These proteins provide a mechanism by which males could manipulate female reproductive behavior. In Drosophila melanogaster, sex peptide (SP, Acp70A) influences egg production, oviposition, and female remating. Inside the female reproductive tract it is attached to sperm and is continuously released for days after mating, stimulating oviposition. However, SP is also known to impose later costs on the female's survival. Yet the direction and magnitude of the SP effects on the female's fitness is largely context dependent, and SP could benefit both males and females if it is a cue of sperm availability. Using a biologically informed model, we show how seminal fluid proteins can synchronize sperm and egg release and reduce the number of unfertilized eggs a female lays. We show that the exhaustion of SP-like protein might be the key signal of this synchronization, which is enabled by the binding to sperm. We stipulate that such synchronization and regulated decrease in oviposition is expected to be common and possibly an understudied general feature of reproduction. Finally, we argue that seminal fluid proteins might be harmful to the female, but the benefits of more efficient reproduction are likely to outweigh the costs.
Polymer electrolytes (PEs) are widely regarded as a promising platform for solid-state batteries (SSBs), offering the potential to simultaneously achieve high energy density with improved safety. However, in current literature, PEs spanning liquid-percolated gels, liquid-assisted quasi-solids, and truly polymer-governed solids are often indiscriminately grouped as solid polymer electrolytes (SPEs), obscuring their distinct ion transport mechanisms, interfacial behaviors, and practical performance constraints, and leading to misleading performance comparisons and unrealistic expectations regarding solid-state operation. Herein, we establish a mechanistic framework that categorizes PEs into gel polymer electrolytes (GPEs), quasi-solid polymer electrolytes (QSPEs), and all-solid polymer electrolytes (ASPEs) based on their dominant ion-solvation environment and transport pathways. By systematically analyzing the ion-transport mechanisms, interfacial behaviors, and performance-limiting features associated with each PE class, we clarify their defining characteristics and mechanism-imposed limitations. Accordingly, we outline category-specific research priorities and highlight the necessity of mechanism-driven materials design, transparent definitions and reporting, and application-relevant benchmarking. This unified Perspective lays a foundation for consistent interpretation, meaningful comparison across PE systems, and more rational materials design toward the advancement of PE-enabled SSBs.
The flavor of rabbit meat has always been a major factor hindering the development of the rabbit industry. One of the main factors affecting the flavor of rabbit meat is intramuscular fat. N6-methyladenosine (m6A) regulates multiple aspects of the physiology of animals. In this study, qRT-PCR and m6A-qPCR were used to identify genes and methylation levels. AAV virus was used as a vector to overexpress genes. To explore the regulatory mechanism of m6A on intramuscular fat in rabbits, we first explored the regulation of the LPL gene of rabbits by m6A at the cellular level using interfering RNA. Subsequently, we further validated the mechanism and explored the regulation of metabolites by LPL genes in living dorsal muscles. The results demonstrate that METTL3 inhibited LPL expression through m6A modification under the recognition of YTHDF2 in adipocytes and muscles. LPL promotes adipocyte differentiation and intramuscular fat deposition. In addition, LPL regulates intramuscular fat deposition through L-Glutamine/multiple pathways and 3-Methyl-L-histidine. This study confirms that m6A can affect the expression of the LPL gene in rabbits, thereby regulating the IMF of rabbit meat by L-Glutamine/multiple pathways and 3-Methyl-L-histidine. This study lays the molecular foundation for cultivating high-quality rabbit meat.
The long-pulse pulsed power facility composed of capacitor banks can serve as a loading method to provide high-speed, high-pressure implosion conditions for solid liners. The implosion process is characterized by high symmetry, repeatability, and ease of diagnostics, enabling the study of physical properties of materials and complex fluid dynamics under extreme conditions. However, previous designs for the load region were optimized primarily for cylindrical convergence structures, which are less adaptable when dealing with non-converging geometries. Moreover, for experiments that require comparing physical differences caused by geometric configurations, it is generally not straightforward to perform the same round of experiments using the same loading method. Therefore, we develop a new structure of load region in FP-2 (facility for Fluid Physics investigations-the second generation) that allows planar and cylindrical dual-loading modes to be carried out simultaneously. By calculating dynamic model combined with full-circuit model and electromagnetic simulation, the structural parameters of flyer plate and cylindrical liner were designed. Based on the loading device combined with laser interferometry diagnostic technology, verification experiments, such as flatness test of large-sized flyer plate, were conducted. The experimental results of planar launch and cylindrical implosion in the same experiment demonstrate the feasibility of simultaneous loading. In addition, the dual-loading method was finally applied to spalling experiment, rebound velocity signals were obtained for two kinds of geometric configurations. Compared with conventional single-mode systems, the new device enhances the efficiency-to-cost ratio of experiments, lays a foundation for further structural optimization to enable configuration comparison studies under identical loading conditions, and provides a dual-loading platform for other physical experiments, such as interfacial instability investigations.
Glycogen Synthase Kinase 3 (GSK3) is a family of highly conserved serine/threonine protein kinases originally identified in animals. Subsequently, homologous proteins with analogous functions were discovered in plants and designated as GSK3-like kinases or SHAGGY-like kinases (SKs). These kinases play pivotal roles in plants, including participation in growth and development, abiotic stress responses, and hormone signaling. However, the bioinformatics characteristics and protein functions of this gene family in foxtail millet remain poorly characterized. This study identified 9 SHAGGY-like kinase gene family members in the foxtail millet, which were classified into four subfamilies based on phylogenetic analysis. The functions of the gene family members were dissected by integrating bioinformatics analysis with quantitative real-time PCR (qRT-PCR). Chromosomal localization analysis revealed uneven distribution of family members across chromosomes, prompting renaming based on their chromosomal order. Analysis of conserved domains, motifs, and gene structures demonstrated high conservation among subfamily members. Transcriptome analysis indicated active expression offoxtail millet SHAGGY-like kinase family members in stems and leaves. Stress treatment experiments showed that most foxtail millet SHAGGY-like kinase genes are responsive to abiotic stresses (drought, cold, NaCl) and exogenous hormones (ABA and MeJA). In summary, the phylogenetic and functional analysis of the foxtail millet SHAGGY-like kinase gene family provides insights into its role in stress responses and lays the foundation for future research aimed at enhancing the stress tolerance of foxtail millet.
Coronaviruses are RNA viruses with high genomic plasticity and with the ability to infect a wide range of animal hosts, including humans. Among them, the highly pathogenic SARS-CoV-2, a betacoronavirus, is the causative agent of severe respiratory infection, and has also been associated with extrapulmonary manifestations, including cardiac involvement. BCoV, another betacoronavirus, is a major pathogen of cattle, primarily affecting the respiratory and enteric systems. Herein, a case of death in a calf with clinical signs consistent with BCoV infection is described. Potential cardiac involvement was further investigated, motivated by evidence of SARS-CoV-2 associated cardiac pathology in humans. Pulmonary and cardiac tissues were subjected to molecular and histological analyses. BCoV RNA was detected in both heart and lungs while immunohistochemical and immunofluorescence analyses confirmed BCoV antigen presence in both tissues. Cardiac detection was mainly observed in the conduction system and the endothelial cells. This study provides the first evidence of cardiac detection of BCoV and lays the foundation for future research on the closely related human coronavirus OC43, contributing to the One Health approach.
Octenyl succinic anhydride (OSA) modification is an effective strategy to improve the emulsifying properties of carbohydrates. OSA-highly branched cyclodextrin (HBCD) with different average degrees of substitution (DS) were prepared in the study. The influence of the modification on the structure, physicochemical properties and emulsifying performance was investigated. Fourier transform infrared spectroscopy and nuclear magnetic resonance hydrogen spectroscopy confirmed the successful grafting of octenyl succinate groups onto HBCD chains, and the substitution reaction occurred mainly at the C6 position of the glucose unit. The introduction of octenyl succinate groups reduced the thermal stability of HBCD while significantly enhancing its hydrophobicity. OSA-HBCD was able to form stable O/W Pickering emulsions, among which the sample with an average DS of 0.028 exhibited the best emulsifying performance. All the OSA-HBCD stabilized emulsions showed shear thinning and viscosity dominant (G″ > G') behavior, and higher average DS corresponded to greater viscoelasticity of the emulsions. The study lays a theoretical groundwork for employing OSA-HBCD as an emulsifier in functional food applications and related fields.
Maize (Zea mays L.) is a globally important crop. Understanding plant height and the underlying dwarfing genes is of critical importance for improving lodging resistance and optimizing plant architecture. In this study, a dwarf mutant designated CM17-2 was obtained by exposing the maize inbred line B73 to carbon ion beam irradiation. The mutant exhibited markedly reduced plant height, internode length, and ear height. Segregation analysis in the F2 population revealed a 3:1 ratio, indicating monogenic recessive inheritance. Using bulked segregant analysis (BSA), the causal locus was mapped to four intervals on chromosomes 2, 4, and 5. Exogenous GA3 treatment rescued the dwarf phenotype and restored plant height to near-wild-type levels, indicating GA sensitivity. Transcriptomic analysis following GA3 treatment identified 3292 differentially expressed genes (DEGs) between CM17-2 and B73. Among these, 1664 GA3-responsive genes were significantly enriched in the plant hormone signal transduction pathway. Integration of BSA-seq and transcriptomic data pinpointed 18 candidate genes on chromosome 4. Through gene annotation, four potential candidate genes likely associated with dwarfism were identified, and the functions of these candidate genes need to be validated in future experiments. The identification of these potential candidate genes lays a theoretical foundation for the subsequent functional analysis, gene cloning, and mechanistic studies of maize dwarfism-related genes.
Aortic stenosis (AS) and left ventricular diastolic dysfunction (LVDD) often coexist in heart failure (HF), but the mechanisms linking them remain unclear. While AS increases afterload and promotes myocardial stiffening, emerging AI-based evidence suggests LVDD can precede the development of AS or progress simultaneously, indicating shared upstream mechanobiological and inflammatory drivers. This study explores the genetic contributors connecting AS and LVDD to identify early molecular markers and convergent pathways in HF. We analyzed Whole Genome Sequence (WGS) and RNA-seq data of the HF patients, generated using their Peripheral Blood Mononuclear Cells (PBMCs) samples. Overall bioinformatics analysis was divided into two modules, 1) gene variant and annotation analysis, and 2) gene expression and enrichment analysis. We utilized our peer review published and open source WGS and RNA-seq pipelines to process Next-Generation Sequence (NGS) data. Furthermore, we performed bioinformatics and statistical analysis to identify genetic variations, expressions, regulation, enrichments, and disease annotations. We identified genetic markers uniquely associated with AS, LVDD, and shared between them. Furthermore, we report genes with significant expression, and functional variations, and discuss their relationship with other cardiovascular diseases (e.g. Vascular and Cardiac Stiffness, Aortic Dissection, Left Atrial Enlargement, Left Ventricular Hypertrophy, Outflow Tract Obstructive Defects, Non-Compaction Coronary Artery Disease, Arrhythmia, Congestive Heart Failure, and Hypertrophic, Dilated, and Ischemic Cardiomyopathy) and non-cardiovascular diseases (non-CVDs) (e.g. Type 1 Diabetes, Diabetic Nephropathy, Skeletal Anomalies, Rheumatoid Arthritis, Atypical Femoral Fractures, Chronic Kidney Disease, Dehydrated Hereditary Stomatocytosis, Schizophrenia, Varicose Veins, High-Altitude Pulmonary Edema, Periodontitis, and Respiratory Disorder) including multiple cancer types (e.g. Breast, Lung, Colorectal, Pancreatic, Hypopharyngeal, Acute Lymphoblastic, and Oral Squamous Cell Carcinomas) and rare genetic disorders (e.g. Hypophosphatasia, Multiple Sclerosis, Campomelic Dysplasia, Lymphatic Malformation). We validated our results through state of science literature, gene-disease annotation databases, and electronic health records. AS and LVDD share both clinical and genomic associations, with overlapping genetic drivers that are enriched in pathways related to inflammation, extracellular matrix remodeling, and vascular stress responses. This work supports the potential of blood-based multi-omics profiling to uncover early, systemic molecular signals of cardiac dysfunction and lays the groundwork for future tissue-specific studies to guide precision diagnosis, risk stratification, and targeted therapeutics in HF.
Protein amyloid fibrils (AFs) have garnered significant attention in the food industry. However, the acid-thermal preparation method has obvious limitations, such as harsh reaction conditions and poor product homogeneity. Therefore, this review aimed to summarize recent research advances in the formation of AFs from various source proteins, with a focus on innovative strategies for optimizing preparation conditions. It also provided an in-depth analysis of the regulatory mechanisms governing the formation of AFs, influenced by environmental factors such as pH, ionic types and strength, heat conditions, and interactions between substances. The kinetic characteristics and morphological evolution of the AFs assembly process were elucidated at the molecular level. The overview of aforementioned formation mechanism lays theoretical foundation for the precise regulation of the functional properties of AFs. Furthermore, thanks to their excellent functional properties, AFs are increasingly applied and play a crucial in fields such as materials science, biomedicine, and food innovation. Therefore, this review presented the latest research trends and application prospects of AFs in these fields. Finally, while comprehensively analyzing the technical advantages, this review also objectively pointed out challenges in recent investigations, including difficulties in large-scale production and insufficient safety assessment, and put suggestions for development suggestions. This review provides valuable theoretical basis and technical reference for the efficient preparation of AFs from edible proteins through modification or environmental regulation strategies. It also holds its significant research value and broad application potential in the development of novel functional materials, personalized food products, and related fields.
Understanding high-level visual processing requires data that capture both fine-grained neuronal activity and large-scale cortical organization. We present the Triple-N dataset, which extends the Natural Scenes Dataset (NSD) framework to macaques by combining functional magnetic resonance imaging with dense Neuropixels recordings in the inferotemporal cortex and early visual areas during the viewing of 1,000 NSD images. Neuropixels probes provide high-resolution population sampling, capturing hundreds of simultaneously isolated units with millisecond temporal precision. Using these data, we show that inferotemporal category-selective regions exhibit robust tuning for their preferred categories, and dense sampling further reveals diverse temporal response patterns and image-dependent latency variations that reflect both intrinsic neuronal properties and stimulus features. Aligning macaque electrophysiology with human NSD functional magnetic resonance imaging demonstrates cross-species correspondences and divergences in representational geometry. Overall, the Triple-N dataset lays a foundation for unifying single-neuron dynamics, cortical representations and cross-species comparisons, helping to shape a more comprehensive understanding of primate visual processing.
Initiating controlled radical polymerizations directly from common functional groups is desirable for synthesizing advanced polymer architectures, such as surface grafts and polymer conjugates. Previously, photocontrolled atom transfer radical polymerization (ATRP) has been performed with alkyl halide initiators, limiting its utility in these advanced polymer applications. In this study, we demonstrate the initiation of photo-ATRP directly from the α-carbon of primary amines. We employ an iridium photocatalyst to trigger radical deamination of redox-activated primary amines, initiating controlled radical polymerization in the presence of an exogenous bromide source under visible light irradiation. The resulting polymers have narrow molar mass distributions (Đ ∼ 1.2-1.3), good agreement between theoretical and experimental molar masses, exceptionally high α-chain-end fidelities (>99%), and active bromide ω-chain-ends. This study lays the groundwork for utilizing non-alkyl halide initiators to initiate well-controlled ATRP polymerizations, opening the door for the use of redox-activated functional groups as ATRP initiators for creating advanced polymer architectures.
Decoy state semi-quantum key distribution (SQKD) has been proposed as a method aimed at significantly enhancing the security and performance of practical semi-quantum key distribution systems. However, the gap between theory and experiment remains open. Herein, we propose the decoy state setting and experimental scheme for the mirror protocol and conduct the proof-of-principle demonstration experiment of the decoy state mirror protocol. The simulation results, obtained under experimental conditions, demonstrate that no key generation rate can be generated under the conditions of the Gottesman-Lo-Lütkenhaus-Preskill (GLLP) and one-decoy state, while valid key generation rates are achievable within a fiber transmission distance of 19.1 km under the weak and vacuum states. Furthermore, it was experimentally verified that a key generation rate of 406 bits/s can be achieved under the combined conditions of weak and vacuum states and a 10 km fiber channel. As the first, to the best of our knowledge, proof-of-principle demonstration of the decoy state mirror protocol, this study lays a foundation for the practical security analysis of SQKD from an experimental perspective and serves as an important starting point for the practical verification in this field.
Maturity Onset Diabetes of the Young (MODY) is a type of genetically inherited diabetes. As it generally arises in early adulthood, it can affect pregnant women with significant neonatal outcomes. Our study aimed to conduct a systematic review on pregnancy management and outcomes of all subtypes of MODY, browsing the Cochrane, EMBASE and PubMed databases to find clinical studies on the topic. They were last searched on June 29, 2025. The inclusion criteria for our review were: published articles, available in their full-text version, about MODY, reporting data about its management during pregnancy and neonatal outcomes, in English language, specifying therapeutic approaches used. The exclusion criteria were: study designs reported below, written in other languages, not present in above mentioned databases. The risk of bias was assessed through the ROBIS tool. Our systematic review analyzed data on 1408 live births of 1183 pregnant women, and found specific guidelines for the management of pregnant women affected with GCK-MODY and HNF1A-MODY, but none on rarer subtypes. Thus, indications on pregnancy management and outcomes of rarer MODYs were identified through an additional literature review, and its findings were summarized in Table 1. Our systematic review was limited by the lack of data on rarer subtypes of MODY and the specific keywords and databases selected. However, our work lays an important step in the direction of much-needed clinical data and recommendations in treating rare MODY subtypes during pregnancy, towards healthy offspring delivery and growth. A further literature review was carried out to identify any data on other subtypes of MODY and their neonatal outcomes, and the findings were summarized in Table 1. No funding was needed to carry out the present systematic review. PROSPERO registration number: CRD420251062633.
Pulsed field ablation (PFA) is a novel, non-thermal method for the treatment of atrial fibrillation (AF). We conducted this study, as part of one of the earliest reports originating from the MENA region, to compare the efficacy of PFA vs. medical therapy alone for the treatment of AF. A retrospective cohort study was conducted in which 62 patients who underwent PFA ablation for AF were compared with a cohort of 62 propensity score-matched patients sourced from the JoFib Registry. Rates of readmission in both arms due to AF recurrence or heart failure exacerbation driven by rapid ventricular response to AF within the period of 1 and 6 months were considered the primary study endpoints. Data were cleaned and analyzed using R 4.5.3. Both cohorts were well matched in terms of age, gender, diabetes mellitus, hypertension, heart failure, left atrial diameter, CHA2DS2-VASc score, ejection fraction, and prior use of antiarrhythmic drugs. Compared with medical therapy, PFA showed significantly lower readmission rates at 1 and 6 months (3.2% and 4.8% vs. 12.9% and 17.7%; p < 0.05). A Kaplan-Meier analysis showed significantly higher freedom from readmission rates in the PFA cohort compared with the JoFib cohort (92% vs. 69%; log-rank p = 0.002). This early report from the MENA region aligns with the global consensus supporting PFA as a treatment for AF and lays the foundation for future larger studies to investigate PFA in this specific population.
Pulmonary arterial hypertension (PAH) is a complex disease characterized by chronically elevated pulmonary arterial pressure, with early onset and progression linked to structural, metabolic, and morphological changes in the pulmonary vasculature. Understanding the interplay between hemodynamics and arterial wall mechanics is essential to capture the pathology of the distal vasculature in PAH. This study aims to develop a data-driven framework that establishes a baseline state of PAH vasculature, incorporating key features of arterial wall constituents, geometry, and their interaction with PAH-specific hemodynamics. Illustrative examples of symmetrically bifurcating arterial trees are used to define representative baseline characteristics of PAH-affected pulmonary arteries. Compared with healthy homeostatic vasculature, the computational results demonstrate pronounced geometric and mechanical alterations: Arterial stiffness increases from approximately 7-10 kPa in healthy arteries to 300-800 kPa in PAH, representing a ~ 40-85 times increase across generations. Because wall thickening is imposed from histological measurements while outer diameter is preserved, the diameter-to-thickness ratio (D/h) decreases from ~ 14 in healthy arteries to ~ 3.8 in PAH, reflecting severe lumen narrowing and medial hypertrophy. In addition, the metabolic energy cost per unit length in PAH is more than double that of healthy arteries when assuming unchanged metabolic consumption per unit volume, whereas enforcing equal total energy cost yields a reduced per-volume metabolic consumption of ~ 450-500 W/m3. These findings suggest that maintaining constant metabolic consumption per unit volume would impose excessive energetic demand on the pulmonary vasculature in PAH, whereas redistribution of metabolic expenditure through altered wall composition may represent a more physiologically plausible adaptation. Furthermore, this framework provides a quantitative baseline state for PAH vasculature and lays the groundwork for future integration of growth-and-remodeling analyses and pharmacological pathway modeling to evaluate treatment response.
Overuse of pesticides and herbicides, such as glyphosate, is a major global concern due to potential negative impacts on human and environmental health. This makes rapid detection of these environmental chemical contaminants necessary for monitoring and mitigation efforts. Our previously developed RNA output sensor activated by a ligand induction (ROSALIND) cell-free biosensor platform enables rapid, point-of-need detection of chemicals through transcription factors that sense chemical targets and activate reporter gene expression in vitro. However, ROSALIND currently relies on known transcription factors that directly bind chemical ligands to transduce a signal, limiting the range of chemicals that can be detected by this technology. Here, we expand ROSALIND to incorporate enzymes that convert chemical targets of interest into detectable compounds. We show a proof-of-principle of this design by engineering ROSALIND to detect the herbicide glyphosate by enzymatically converting it to glyoxylate, which binds the transcription factor AllR to activate the gene expression. This work develops a workflow for expanding in vitro transcription biosensors by interfacing them with enzymatic conversion pathways and lays the groundwork for developing functional glyphosate sensors for real-world applications.