Search Results (8,039)

Translation of mRNAs is a tightly regulated process in gene expression. In mRNA, the 5′ untranslated region (5′ UTR) controls ribosome recruitment and frequently contains structured elements that modulate translation efficacy. This study investigates stable structural motifs within the 5′ UTR of DDX23 mRNA, encoding a protein relevant for anticancer therapy, as potential regulators and targets for antisense oligonucleotides (ASOs). Despite bioinformatic predictions and transcriptomic validations suggesting RNA G-quadruplex (rG4) formation, comprehensive structural analysis using a light-up assay and CD, UV, and NMR spectroscopy revealed that most putative rG4-forming sequences do not fold into stable rG4 structures, although one of them exists in an equilibrium between rG4 and an alternative, likely hairpin, conformation. Reporter assays using a robust G4 stabilizer also argue against the significant regulatory role of rG4s in DDX23 mRNA translation. Instead, we identified and characterized a stable hairpin structure with potential regulatory function. Based on these findings, we designed fully locked nucleic acid (LNA)-modified ASOs to target this hairpin and regions flanking the upstream open reading frame (uORF) and start codon of the coding sequence. A reporter assay demonstrated that cap-proximal targeting achieved robust translation inhibition up to 80%. In contrast, targeting the efficiently translated uORF was ineffective, presumably due to steric hindrances from the ribosomal complex. The study yields crucial design principles for translation-regulating ASOs: avoid targeting regions shielded by efficient uORF translation and carefully tune ASO-RNA duplex stability to surpass endogenous structures without disrupting regulatory mechanisms. These findings provide insights into the regulation of DDX23 expression and establish a framework for developing ASO-based therapeutics with broad implications for mRNA targeting in anticancer applications. Full article

The global challenge of food insecurity requires innovative approaches for sustainable food production and waste valorization. This study investigates the valorization of oat hulls, an abundant lignocellulosic by-product from oat manufacturing, by solid-state fermentation using edible filamentous fungi. Oat hulls sourced from oatmeal industrial side-streams were used as the sole substrate in co-cultures of Neurospora intermedia and Rhizopus oryzae. The fermentation process was optimized and upscaled, with fungal growth monitored via CO₂ efflux and modeled to assess substrate utilization. Comprehensive analyses revealed a significant increase in protein concentration (p < 0.05) in the fermented oat hulls compared to the non-fermented controls. The resulting product was successfully incorporated into granola bars, which underwent sensory evaluation and received positive feedback, demonstrating its potential as a value-added food ingredient. These findings highlight the feasibility of using edible fungi to upcycle cereal processing by-products into nutritionally enhanced alternative protein sources, supporting both food system sustainability and circular bioeconomy objectives. Full article

This study aimed to explore innovative sorbent materials for the remediation of contaminated marine environments, with a focus on metal removal from seawater. Adsorption tests were carried out to evaluate the performance of single-cell proteins (SCPs), a protein-rich biomass derived from industrial by-products, in comparison with commercial activated carbon (AC). Given the increasing need for sustainable and effective approaches in sediment remediation and water treatment, identifying alternatives to conventional sorbents is of particular relevance. Results showed that SCPs exhibited higher affinity for Cr than for Zn, while multi-metal solutions improved adsorption, suggesting synergistic interactions possibly linked to surface charge effects and ternary complex formation. Importantly, SCPs demonstrated competitive and, in some cases, superior performance compared to AC, highlighting their potential as an innovative and sustainable material. Moreover, when the absorbent materials were combined, SCP and AC mixes outperformed both the individual adsorbents and the expected additive efficiencies, achieving significantly higher removal yields for both metals, particularly at low concentrations. Overall, these findings suggest that SCPs, alone or in combination with AC, represent a promising strategy for the removal of heavy metals from marine systems, offering new opportunities for the treatment of contaminated sediments and seawater. Full article

Quinoa (Chenopodium quinoa Willd) is a nutrient-rich multipurpose crop. Its grains are used as a cereal, green leaves as a vegetable for humans, and the whole green plant as an alternate forage for livestock. Recently, whole-plant quinoa forage has been evaluated in several countries in Asia and Europe for its potential use as an alternative forage for livestock; however, this has not been performed in the United States. We investigated forage yield and related agronomic traits, nutritional composition, and feed quality-related traits in 60-day-old quinoa whole plants of four quinoa lines over a two-year period. The goal was to evaluate the feasibility of quinoa forage production in Missouri, a drought-prone midwestern state of the USA. Morphological traits (height and fresh and dry weight per plant), chemical composition (fiber contents), and nutritive quality (digestible nutrient contents) of forages were affected by quinoa genotype and year of planting. The crude protein content of quinoa forage averaged 16.23% and fiber 22.08%, which was similar to the values reported in Asia and Europe, but was slightly lower than that of alfalfa. Calcium (1.26%) and phosphorus (0.47% dry weight) were significantly higher than those reported in published quinoa forage results and are comparable to those in published alfalfa minerals. Lysine (0.98%) and methionine (0.25%) were higher than the published results for quinoa and alfalfa. Neutral detergent fiber (34.10%) and acid detergent fiber (25.01%) were lower than those of alfalfa, indicating better digestibility of the quinoa forage. The calculated digestible dry matter (69.40%), dry matter intake (3.56%), relative food value (192%), and total digestible nutrient (70.33%) were higher than those of alfalfa and comparable with published results for quinoa forage. Our preliminary results indicate that the quinoa lines evaluated in this study have excellent potential to be used as a non-traditional alternative forage, especially in environmentally stressed areas where the production of other forage crops is limited. Further research should explore the full multipurpose benefits of quinoa, including its use as grains, leafy green, and whole-plant forage. Full article

Hematophagous leeches rely on a broad diversity of bioactive factors that interfere with the host’s defense systems to secure a successful blood meal. The most prominent examples of such factors are the antithrombotics that address either the primary hemostasis (the platelet aggregation) or the secondary hemostasis (the blood coagulation). Whereas the inhibitors of platelet aggregation mainly cause continuous blood flow, coagulation inhibitors mainly keep the blood fluid within the stomach of the leech. The critical dependency of hematophagous leeches on the accurate action of all antithrombotic factors may explain the presence of multiple genes for each type of these factors that is regularly observed in leech genomes. The genome of Hirudinaria manillensis Lesson, 1842, the Asian buffalo leech, contains five individual genes that encode variants of the coagulation inhibitor hirudin. However, no genes that encode putative decorsins, the archetype of leech-derived inhibitors of platelet aggregation, have been described to date. Here, we report the identification of one monomeric and four multimeric decorsin encoding genes in the genome of H. manillensis. A selection of the putative decorsins was expressed as recombinant proteins, purified and functionally characterized. All but one of these putative decorsins displayed platelet aggregation-inhibitory potencies. Furthermore, we provide a mathematical calculation based on the mechanism of alternative pre-mRNA splicing that illustrates the potential to generate an enormous variety of different factors from one single multimeric ornatin gene. Full article

Background and Objectives: Age-related macular degeneration (AMD) is an age-associated retinal disorder characterized by blood–retinal barrier (BRB) breakdown and pathological angiogenesis, leading to vascular leakage. The intravitreal administration of anti-VEGF agents remains the most effective treatment for neovascular AMD. However, repetitive intravitreal injections have risks, causing side effects such as cataracts, bleeding, retina damage, and, in severe cases, post-injection endophthalmitis. Hence, the development of innovative drug delivery systems is essential to minimize the risks and discomfort associated with intravitreal injections. Materials and Methods: We developed a microemulsion (ME)-based topical drug delivery system incorporating α-linolenic acid (ALA). In brief, pseudo-ternary phase diagrams were constructed by the water titration method using different combinations of surfactants and cosurfactants (S_mix-Cremophor RH 40: Span 80: Transcutol P in ratios of 1:1.05, 1:1:1, 1:1:1.5) containing ALA as the oil phase. Three blank microemulsions (ME1, ME2, and ME3) were prepared and characterized based on the optimized pseudo-ternary phase equilibrium with a S_mix ratio of 1:1:1. Results: ME3, with an average particle size of 38.59 nm, was selected as the optimized formulation for developing drug-loaded ME containing Fenofibrate, Axitinib, and Sirolimus. The drug-loaded ME showed particle size (46.94–56.39 nm) and in vitro release displayed sustained and longer time drug release for 240 h. The irritation and antiangiogenic activities were evaluated using the hen’s egg chorioallantoic membrane (HET-CAM) assay employing the optimized ME loaded with each drug. Among the three drug-loaded ME, the Sirolimus ME showed a reduction in blood vessel sprouting in the HET-CAM assay, indicating strong antiangiogenic activity. Treatment with the optimized blank ME and Sirolimus ME significantly (p < 0.05) reduced COX-2 protein expression in LPS-stimulated RAW 264.7 cells, suggesting their potential anti-inflammatory effects. Conclusions: Overall, we suggest that the α-linolenic acid-based Sirolimus microemulsion may serve as a promising topical therapeutic approach for managing AMD and offering a potential alternative to invasive intravitreal injections. Full article

An eight-week study was conducted to evaluate the effects of dietary marine protein hydrolysates as fish meal replacements in low-fish-meal diets on the growth performance, feed utilization, and health status of Asian seabass (Lates calcarifer). The high-fish-meal (HFM) diet contained 25% fish meal, while the low-fish-meal (LFM) diet replaced 60% of the fish meal with soybean meal. Three experimental diets were formulated by supplementing the LFM diet with 5% tuna hydrolysate (TH), 2% shrimp hydrolysate (SH), and 5% salmon silage (SS), each replacing an equivalent amount of fish meal. These diets were designated as LFM + TH, LFM + SH, and LFM + SS, respectively. The results showed that the LFM + TH diet significantly improved the percentage of weight gain, average daily growth, specific growth rate, protein efficiency ratio, and feed conversion ratio compared to the LFM diet (p < 0.05), without negatively affecting feed intake or metabolic markers. Histological analysis revealed improved villus length and goblet cell count in the intestine, indicating better nutrient absorption (p < 0.05). However, no significant differences were observed in hematological and immunological parameters, blood plasma metabolic markers, or carcass proximate composition (p > 0.05). Furthermore, the LFM + TH diet exhibited superior survival rates under ammonia stress, highlighting its potential to enhance stress tolerance. These findings suggest that marine protein hydrolysates, particularly 5%TH, can serve as a sustainable and efficient alternative to fish meal protein in diets with up to 60% in soybean meal compensation, promoting better growth and survival in Asian seabass. Full article

This study systematically explored the regulatory mechanisms of uterine function across four reproductive stages: sexual maturity sow (SMS), low-yield sow (LYS), high-yield sow (HYS), and culled sow (CS) in Large White (LW) pigs through integrated transcriptomic, proteomic, and metabolomic analyses. Twelve healthy LW sows were selected, and uterine tissues were collected for multi-omics detection. Combined with bioinformatics analysis, molecular regulatory networks were constructed. Results showed that transcriptomics identified 12 types of alternative splicing and 1243 novel genes, which were enriched in energy metabolism and signal transduction pathways. Proteomics revealed 430 differentially co-expressed proteins, indicating high protein synthesis activity in the SMS stage and extracellular inflammatory characteristics in the CS stage. Metabolomics detected numerous differential metabolites, among which XTP and DHA ethyl ester were associated with high fecundity and aging, respectively. Integrated multi-omics analysis identified hub genes such as PLA2G4A, which influence reproductive performance by regulating inflammatory and metabolic balance, and clarified stage-specific “gene–protein–metabolite” modules. This study provides a molecular map for understanding dynamic changes in uterine function in Large White pigs and offers a theoretical basis for optimizing reproductive lifespan and breeding strategies. Full article

The emergence of multidrug-resistant Shiga toxin-producing Escherichia coli (STEC) poses a major challenge to public health and necessitates the development of alternative antimicrobial strategies. This study aimed to isolate and characterize five lytic bacteriophages belonging to the genus Mosigvirus and evaluate their potential as biocontrol against MDR STEC strains and their biofilms. The five bacteriophages, designated vB_EcoM-pJBB (ΦB), vB_EcoM-pJBC (ΦC), vB_EcoM-pJBJ (ΦJ), vB_EcoM-pJBK (ΦK), and vB_EcoM-pJBL (ΦL), were isolated from sewage treatment plant samples using STEC ATCC 43895 as host. Biological characterization included host range determination against 19 MDR STEC strains, one-step growth analysis, environmental stability assays, bacteriolytic activity assessment, and antibiofilm efficacy testing. Whole-genome sequencing and phylogenetic analyses were performed to determine genomic features and taxonomic classification. The phages demonstrated varying infectious capacities, lysing between six and 12 strains, with ΦL exhibiting the broadest spectrum of activity. All phages showed MOI-independent antibiofilm activity, preventing biofilm formation by approximately 70% and disrupting pre-formed biofilms by up to 80.3%. Genomic analysis revealed the absence of lysogeny markers, virulence factors, and antimicrobial resistance genes, while identifying putative depolymerase genes associated with tail fiber proteins. Phylogenetic analysis confirmed the taxonomic position of these phages within the Mosigvirus genus in the Straboviridae family. Our findings indicate that the newly identified Mosigvirus phages are promising candidates for phage-based biocontrol applications. Full article

Background: The immunogenicity of polysaccharide conjugate vaccines is critically influenced by the choice of carrier protein, which promotes a T-cell-dependent immune response mechanism leading to strong antibody production. In this study, the cholera toxin B subunit (CTB), a non-toxic pentameric protein, was evaluated as a novel carrier protein for pneumococcal polysaccharide antigens. Methods: Recombinant CTB was produced in Escherichia coli and purified using scalable chromatographic methods. Pneumococcal polysaccharides from serotypes 7F, 22F, and 33F were chemically activated with CDAP and conjugated to CTB. Results: The resulting glycoconjugates were characterized by SEC-MALS, confirming successful conjugation, high molecular weights, consistent polysaccharide-to-protein ratios, and acceptable endotoxin levels. Immunogenicity was assessed in rabbits following immunization with alum-adjuvanted formulations. Results: Robust IgG responses were elicited by all CTB-based conjugates, with antibody levels found to be comparable to those induced by CRM197 conjugates, demonstrating the potential of CTB as a promising alternative for the next generation of conjugate vaccines. Full article

Plant-based proteins offer sustainable alternatives to animal sources, yet their lower digestibility remains a critical barrier to widespread applications. Current digestibility assessment methods require days of analysis and gram-scale samples, creating significant bottlenecks in protein optimization workflows. This study developed an ensembled deep learning framework that transforms digestibility prediction from a resource-intensive process to a rapid, minimal-sample assessment. By systematically characterizing 23 diverse plant protein isolates across multiple physicochemical dimensions, we trained a feedforward neural network based on augmented data. Our model identified α-helix content, random coil content, and solubility as key digestibility indicators. This insight enabled the construction of a streamlined three-feature model that reduced assessment time by 80% while requiring only one-hundredth of standard sample amounts. When validated against independent published datasets, the model achieved rational prediction accuracy, with an R² = 0.91. These findings establish a transformative framework for accelerating plant protein development, enabling rapid screening of novel sources and targeted modification strategies to enhance nutritional bioavailability, ultimately advancing sustainable food system transitions. Full article

Background: Inflammation is a common reason for dogs to present to veterinary clinics. Early diagnosis of systemic inflammation is important. Acute phase proteins, like C-reactive protein, are useful but not specific to infection. In human medicine, the intensive care infection score (ICIS) offers a faster, cost-effective alternative using advanced hematological parameters. While ICIS is not available for veterinary use, some components (e.g., neutrophil side fluorescent light) can be measured using analyzers like the Sysmex XN-1000V. Objectives: This study aimed to establish a control group of healthy dogs for the novel parameters neutrophil side fluorescent light (NE-SFL), neutrophil side scattered light (NE-SSC), and neutrophil forward scattered light (NE-FSC) and assess their utility in detecting inflammation in diseases such as sepsis, pyometra, steroid-responsive meningitis-arteritis (SRMA), and idiopathic epilepsy. Methods & Results: Value ranges were calculated based on 21 healthy dogs. Compared to controls, NE-SFL levels were significantly elevated in sepsis, pyometra, and SRMA, while NE-SSC was only elevated in sepsis and pyometra and NE-FSC only in sepsis. No increases were observed in idiopathic epilepsy. Manual gating of the white blood cell differential scattergram was necessary in samples showing high neutrophil toxicity and the presence of bands. Conclusion: NE-SFL and NE-SSC, obtainable from routine complete blood count, may serve as novel, accessible markers for inflammation in dogs. Further research is needed to validate their broader diagnostic use. Full article

Climate change and socio-economic transformation increasingly challenge the stability of China’s food supply. This study aims to optimize grain crop layouts by integrating natural suitability and nutritional supply within a unified analytical framework. Using the MaxEnt model incorporating bioclimatic, topographic, and soil variables, we simulated the natural suitability of major grain crops and compared it with actual planting patterns based on the SPAM dataset. Results revealed substantial spatial discrepancies between actual and suitable distributions, with national planting diversity index increasing by 26.42% (from 0.53 to 0.67) under suitable conditions. Wheat and maize are most suited to northern China, rice and tuber crops to southern regions, while soybean performs optimally in the northeast. Nutrient supply potential also improved substantially under the suitable scenario, with energy, protein, fat, and carbohydrate increasing by 56.9 × 10⁸ KJ, 77.2 × 10⁶ g, 23.3 × 10⁶ g, and 48.6 × 10⁶ g per million people, respectively. Among alternative structures, maize-soybean and maize-based planting structures better aligned with both natural adaptability and nutritional balance (e.g., in Inner Mongolia and Heilongjiang), whereas rice-based structure showed weaker correspondence (e.g., in Shanghai). These findings demonstrate that naturally adaptive optimization can enhance both environmental compatibility and nutritional adequacy, providing scientific guidance for developing climate-resilient and nutrition-oriented crop layout strategies in China. Full article

Polo-like kinase 1 (PLK1) is a serine/threonine kinase that orchestrates multiple critical events during mitosis, including centrosome maturation, spindle assembly, kinetochore–microtubule attachment, and cytokinesis. Dysregulation and overexpression of PLK1 are frequently observed in various cancers, correlating with increased proliferation, metastatic potential, and poor prognosis, which highlights its potential as a therapeutic target. Traditional small-molecule inhibitors have predominantly focused on the ATP-binding site of the N-terminal kinase domain, effectively inducing mitotic arrest and apoptosis in tumor cells; however, these compounds often suffer from limited selectivity and off-target toxicity. The C-terminal Polo-box domain (PBD), responsible for substrate recognition and subcellular localization, has emerged as an alternative and highly selective target for inhibitor design, enabling the disruption of protein–protein interactions critical for PLK1 function. Here, we present a comprehensive review demonstrating the potential inhibition of several compounds against PLK1. This work establishes a foundation for future preclinical development of small molecule-based therapeutics against PLK1-dependent malignancies. Full article

Human Epidermal Growth Factor Receptor 2 (HER2) is a key therapeutic target in breast cancer. However, the application of existing anti-HER2 antibody drugs is limited by such issues as large molecular weight and poor stability. In this study, a series of small protein minibinders targeting HER2 domain IV were de novo designed using the RFdiffusion method. Candidate molecules were selected through a combination of ProteinMPNN and AlphaFold2 screening, and their binding capabilities were further evaluated using Escherichia coli surface display coupled with flow cytometry analysis. By integrating molecular dynamics simulations, confocal fluorescence imaging, and isothermal titration calorimetry (ITC) experiments, a highly efficient minibinder (0_703_6) with nanomolar affinity and a smaller molecular size was finally identified. Compared with the existing drug molecules, the identified minibinder exhibited approximately threefold higher affinity and a threefold reduction in molecular size. This study provides strong support for the development of novel, stable, and easily expressible HER2-targeted therapeutic molecules and also offers new insights into the rapid development of robust breast cancer drugs that may serve as ideal alternatives to monoclonal antibodies. Full article