Circadian rhythms orchestrate physiological processes, including immune function, across a 24-h cycle. This study investigates the temporal distribution of immune cell populations in healthy mice entrained to a 12:12-h light-dark cycle. Inguinal lymph node (iLN) samples were collected at zeitgeber times (ZT) 0 (lights on) and ZT12 (lights off) to assess immune cell composition. A significantly higher proportion of natural killer T (NKT) cells and neutrophils was observed at ZT0 compared to ZT12, while dendritic cells, macrophages, and natural killer (NK) cells showed no significant temporal variation. Additionally, adaptive immune cells, particularly Programmed cell Death protein 1+ (PD1+) Cluster of differentiation 4+ (CD4+) and PD1+ Cluster of differentiation 8+ (CD8+) Tcells, were more abundant during the light phase. These findings suggest a diurnal pattern in immune readiness, with implications for optimizing immunotherapeutic interventions based on circadian timing.

Endothelial-to-mesenchymal transition (EndoMT), a widely recognized biological process leading to abnormal endothelial function, has been implicated in various cardiovascular pathologies. DEAD-box proteins represent the largest family of RNA helicases associated with multiple physiological and pathophysiological processes; however, their role in the homeostasis of endothelial cells (ECs) remains largely unexplored. Here, we show that the levels of DEAD-box protein 3 X-linked (DDX3X), a DEAD-box RNA helicase protein, were significantly increased during EC transition in vivo and in vitro. DDX3X overexpression promoted EndoMT as well as endothelial dysfunction and inflammation, whereas its downregulation effectively inhibited this transition in ECs. Mechanistically, elevated DDX3X resulted in downregulation of bone morphogenetic protein receptor type 2 (BMPR2), a protein that is pivotal for maintaining endothelial homeostasis and function. Furthermore, our co-immunoprecipitation assays demonstrated a molecular interplay between DDX3X and BMPR2. Importantly, DDX3X was shown to promote the lysosomal degradation of BMPR2, thereby interrupting its downstream signal transduction. These findings identify DDX3X as a novel regulator of EndoMT by modulating BMPR2 signaling.

Intracellular Ca2+ regulates insulin secretion from pancreatic β-cells and is influenced by cannabinoid signaling. However, the hydrophobicity and complex pharmacology of cannabinoid ligands prevent precision receptor targeting, limiting our understanding of their roles in modulating insulin release. Here, we use fluorescent Ca2+ imaging to examine how the light-activatable CB2 receptor agonist azo-HU308 modulates Ca2+ dynamics in INS-1 β-cells. UV-A photoactivation of azo-HU308 triggered robust, repeatable Ca2+ transients, and pharmacological profiling revealed this effect was independent of CB2 receptor activation but was instead mediated by extracellular Ca2+ influx through TRPC channels. These findings position azo-HU308 as a novel optical tool for controlling β-cell Ca2+ levels and highlight a non-GPCR pathway by which synthetic cannabinoids can modulate Ca2+ dynamics in excitable cells.

The ability of Acinetobacter baumannii to form biofilm is correlated with its antimicrobial resistance. The identification of antimicrobial drugs acting on biofilm is crucial to develop effective therapies. Previously, we determined that a chionodracine-derived peptide, KHS-Cnd, was able to impair A. baumannii biofilm formation. Here, to investigate the physiological changes underlying this activity, extracellular metabolite profiles of four A.baumannii strains were analyzed by NMR during biofilm formation in the presence of KHS-Cnd. Metabolites involved in biofilm energy metabolism were found extracellularly after KHS-Cnd treatment. Significantly altered pathways were associated with glyoxylate/dicarboxylate and branched-chain/aromatic amino acid metabolism. Overall, differences in extracellular metabolites reflect modifications of biofilm metabolism due to peptide treatment, thus highlighting its therapeutic potential against A.baumannii biofilm-sustained infections.

The aim of this study was to address the lack of protocols for nuclear magnetic resonance (NMR)-based metabolomics in the agri-food sector by providing a reproducible workflow for the preparation and analysis of water-soluble metabolite fractions. These fractions, rich in primary metabolites such as sugars, amino acids, and organic acids, are key to assessing agri-food products' composition, quality, as well as to monitor their manufacturing and development processes. The protocol differentiates solid and liquid matrices, optimizing extraction procedures accordingly. Representative agri-food products-strawberry leaves (solid) and wine (liquid)-were analyzed to demonstrate the method's versatility and applicability. Key steps include tailored sample preparation, optimization of NMR acquisition, and spectral quality control, ensuring high data quality and reproducibility. The proposed workflow enhances reproducibility across agri-food metabolomics studies and facilitates integration into broader food quality, traceability, and safety frameworks.

The rapid shift to online and blended learning in higher education has led to the development and use of digital tools that support student engagement and learning outcomes. This systematic review examines the effectiveness of these digital tools across various disciplines in higher education, focussing on factors that promote or hinder student engagement. A criteria-based comprehensive systematic search of three databases (Scopus, Web of Science and ProQuest, last date of enquiry 20 August 2024) identified 25 studies, inclusion criteria focussing on primary studies describing and evaluating interactive digital tools designed to enhance learning and/or assessment in higher education. Papers were analysed for bias using JBI checklists, and the papers' findings were analysed using a thematic analysis approach. Analysis of the papers uncovered four key design features that foster engagement with effective digital tools: interactivity, ease of use, immediate feedback and personalised learning experiences. Based on these findings, this review proposes a cyclic model for designing digital tools, emphasising an initial needs analysis, integration with course content, active engagement of students and educators, and ongoing refinement based on feedback. This model offers actionable guidelines for educators and institutions aiming to optimise digital tool development in higher education. The papers identified were typically short-term studies, on specific cohorts of students, and more long-term studies of the impact of digital resources are needed to determine long-term learning gain. The systematic review underscores practical strategies for leveraging digital tools to promote active, self-directed learning by focussing on evidence-based principles.

The evolutionarily conserved gene naked cuticle (nkd) has been gone through studies in both invertebrate and mammalian model systems. Nkd proteins play an essential role in the development of Drosophila as a negative-feedback regulator for Wingless signaling. In this research study, we showcase the multifaceted functions of Nkd in Drosophila wing development. The disturbance of nkd dosage genetically disrupts multiple biological processes in the larval stage and the morphologies of adult wings. Our results of high-throughput sequencing indicate that nkd may have profound effects on Drosophila wing development involving more essential signaling pathways. Nkd may serve as a potential integrator of multiple signaling pathways during wing development, extending its functional repertoire beyond simple Wg inhibition.

Senior research thesis courses are a hallmark feature of many undergraduate science programs, with several documented benefits, including the development of research skills and scientific identity alongside career exploration. In this study, we investigated how the senior research thesis experience is perceived by distinct student populations. We surveyed undergraduate students from two programs at a mid-sized, research-intensive university: Biochemistry, a basic science-focused program, and Biomedical Discovery and Commercialization, a hybrid program combining science and business. Both groups were enrolled in identical fourth-year laboratory-based thesis courses. Our analysis measured the impact of the thesis experience on students' scientific inquiry skills and beliefs; furthermore, we examined how these changes influenced their professional socialization as researchers and their postdegree career goals. Our findings suggest that completing a senior research thesis increased students' perception of their research-related skills, regardless of program enrollment. While there were fewer significant changes regarding student epistemological beliefs around scientific research, qualitative and quantitative measures support the idea that students have developed a more positive perception of failure and resilience within research. Additionally, while students within the science-business hybrid program experienced no significant changes in career goals, completion of a senior research thesis had a significant impact on students within the science-based program. Overall, our results demonstrate that laboratory-based thesis courses can have a notable effect on developing student research skills, beliefs about scientific research, and career goals, and that these effects vary based on the student population.