Fucosylated human milk oligosaccharides are valuable targets for microbial biomanufacturing. Here we engineered the probiotic Escherichia coli Nissle 1917 as a versatile chassis for synthesizing both 2′-fucosyllactose and 3-fucosyllactose. We implemented lactose preservation and blocked colanic acid precursor consumption while reinforcing the de novo GDP-L-fucose pathway. This core module was coupled with either WbgL or FutM2 to drive terminal fucosylation. Initial screening yielded a 2′-fucosyllactose titer of approximately 1.21 g/L. Integrating an additional chromosomal wbgL cassette increased the titer to approximately 1.58 g/L, whereas subsequent genomic modifications did not further improve the 2′-FL titer. Cultivation with 30 g/L glycerol and 5 g/L yeast extract significantly improved productivity, enabling the strain to produce approximately 7.71 g/L of 2′-fucosyllactose in shake-flask cultures. Conversely, the FutM2 strain produced 0.38 g/L 3-fucosyllactose without yeast extract and nutrient supplementation unexpectedly suppressed product accumulation despite promoting biomass formation. The work provides a probiotic-chassis proof of concept and identifies product-module physiology as a priority for further engineering.
The success of COVID-19 mRNA vaccines has established mRNA as a powerful therapeutic platform. Key advantages of mRNA drugs include transient expression, rapid scalability, and universal protein-encoding capability. This review focuses on the application of mRNA therapies across a broad spectrum of diseases, including cancer, infectious diseases, genetic disorders, autoimmune diseases, and cardiovascular conditions. It systematically summarizes the latest progress in mRNA drug development, covering three major therapeutic paradigms: mRNA vaccines, protein replacement therapy, and mRNA-based gene editing. Emerging approaches, such as artificial intelligence tools for mRNA therapeutic design and advanced delivery systems, are also discussed. By integrating foundational mechanisms, engineering strategies, and translational examples, this review provides a comprehensive overview of the current landscape and future directions of mRNA-based therapeutics.
Various biotic indices have been used to assess the condition of aquatic macroinvertebrate communities in agricultural watersheds. We collected 171 samples of benthic macroinvertebrates from 57 stream sites on three forks of an agricultural watershed in southeastern Minnesota, USA, and compared the abilities of various diversity, taxa richness, and multi-metric indices to evaluate invertebrate community health and/or level of impairment. Total invertebrate abundance (sample count), total taxa richness, Ephemeroptera-Plecoptera-Trichoptera (EPT) taxa richness, Simpson diversity, Shannon diversity, a regional benthic index of biotic integrity (BIBI), and the national Izaak Walton League’s Save Our Streams (SOS) rating score were all significantly correlated with one another. Total taxa richness, BIBI score, and SOS score displayed nearly identical influences within a principal components analysis (PCA) ordination. Only total abundance and the two diversities differed significantly among stream forks. When the distributions of sites among rating categories (poor or very poor to excellent) were compared for the various indices, nearly all indices rated community health differently from one another. In order from highest ratings to lowest ratings, the indices were: Simpson diversity, Shannon diversity, SOS score, total taxa richness, EPT taxa richness, and BIBI score. Taxa richness measures and the multi-metric BIBI appeared to rate stream health more realistically (i.e., mostly fair or poor site ratings with relatively few good to excellent) than either of the diversity indices or the SOS score (many good, very good, and excellent site ratings), suggesting that the choice of biotic index can significantly influence evaluations of stream communities in an agricultural watershed.
Underwater threats represent an increasing challenge to maritime safety and port security, demanding advanced solutions beyond traditional monitoring and inspection methods. This paper presents the development and field testing of a 5G-enabled Unmanned Surface Vehicle (USV) designed to enhance situational awareness and operational resilience against underwater risks in port environments. The study was conducted at the Port of Valencia, one of Europe’s most advanced smart ports equipped with a 5G Standalone (SA) network. By combining high-speed, low-latency communications with sensing technologies (including sonar modules, optical cameras, and environmental sensors), the proposed system enables real-time data exchange, remote operation, and continuous monitoring across surface and subsurface domains. Experimental trials validated the USV’s ability to detect submerged objects, support emergency response, and perform inspection and environmental monitoring tasks under stable 5G connectivity. These results demonstrate how the integration of unmanned surface platforms with next-generation communication networks can significantly strengthen the capacity to detect, assess, and mitigate underwater threats under teleoperated conditions.
Eggshell powder is a sustainable calcium carbonate-based material with potential as a feedstock for binder jetting additive manufacturing. However, its irregular particles exhibit poor flowability and powder spreading characteristics. Although spray freeze drying granulation has previously been shown to improve the properties of eggshell powder, the influence of process variables on the resulting granules remains insufficiently understood. This study investigates the effects of spraying pressure and slurry feed rate on the properties of granulated eggshell powder. One-factor-at-a-time experiments were conducted to identify feasible ranges of these process variables, and a full factorial design of experiments with two replications was employed to evaluate the main and interaction effects of spraying pressure and slurry feed rate on particle size and flowability of granulated eggshell powder. Spraying pressure had a statistically significant effect on particle size, whereas slurry feed rate did not significantly affect particle size within the investigated range. Increasing spraying pressure resulted in smaller granules. Powder flowability was evaluated using both the Hausner ratio and repose angle. Both measures showed a significant interaction between spraying pressure and slurry feed rate, indicating that powder flowability depends on the combined effects of these two variables. The findings suggest that spraying pressure is the key variable governing particle size, while the combined influence of spraying pressure and slurry feed rate must be considered when controlling powder flowability. The resulting process–property relationships provide a basis for selecting spray freeze drying conditions for converting eggshell waste into more suitable granulated feedstock for binder jetting additive manufacturing.
