Abstract: The volatile compounds(VOCs) evolution of wild ginseng (WG) across growth years is not a unidirectional process but a divergent remodeling of the chemical fingerprint. In this study, HS-GC-IMS combined with chemometrics was employed to characterize the dynamic changes of VOCs in WG at four growth stages(10, 15, 20, and 25 years; n≥15 per group). A total of 68 VOCs were tentatively identified and semi-quantified, encompassing terpenes, aldehydes, ketones, alcohols, esters, pyrazines, and other classes. Among them, terpenes and pyrazines exhibited the most pronounced directional trends, marking the divergent evolution: terpenes such as camphene, (E,E)-α-farnesene, and β-ionone accumulated progressively (increases of 242%, 74.6%, and 93.4% from 10 to 25 years, respectively), whereas pyrazines including 2,3,5-trimethylpyrazine and 2,5-dimethylpyrazine declined continuously (decreases of 58.2% and 53.3%, respectively). In contrast, the majority of compounds(66%) displayed non-monotonic patterns, including stage-specific metabolic peaks and environmentally driven fluctuations, underscoring the complexity of this divergent evolution. Partial least squares-discriminant analysis (PLS-DA) effectively distinguished samples across growth years (R²Y=0.997, Q²=0.993), with a 200-times permutation test confirming no overfitting(R²=0.136,Q² intercept=−0.505). Twenty-nine differential compounds with variable importance in projection (VIP)>1 were identified as potential chemical markers, and a multi-marker combinatorial system was tentatively established for discriminating three growth stages (10–15, 15–20, and 20–25 years). These findings provide chemical evidence that WG flavor quality evolves divergently over time, suggesting that VOCs fingerprint could serve as a supplementary tool for growth-year assessment, particularly for "high-quality but poor-shape" specimens that are undervalued by traditional morphology-based methods.

Abstract: UBE3A is a dosage-sensitive HECT E3 ubiquitin ligase whose neuronal expression is shaped by genomic imprinting at the 15q11.2–q13 locus. Opposite directions of UBE3A dosage imbalance contribute to distinct neurodevelopmental phenotypes: loss of maternal UBE3A underlies Angelman syndrome, whereas maternally derived duplications involving UBE3A contribute to Dup15q-associated syndromic autism phenotypes. This review synthesizes evidence across molecular architecture, isoform biology, neuronal imprinting, synaptic regulation, circuit excitability, and therapeutic development. The central argument is that UBE3A should not be interpreted as a general explanation for autism, but as a mechanistically informative model for a defined subset of neurodevelopmental disorders in which parent-of-origin effects and copy-number state are central. In Angelman syndrome, UBE3A loss disrupts proteostasis, synaptic plasticity, inhibitory circuit function, and neuronal excitability through distributed rather than single-pathway mechanisms. In maternal Dup15q syndrome, increased UBE3A dosage is strongly implicated in neuronal and synaptic abnormalities, although interval-wide dosage effects also contribute. Therapeutically, the direction of dosage change creates opposite translational requirements: restoration or paternal reactivation in Angelman syndrome versus dosage normalization in Dup15q-associated gain-of-function states. A dosage-directionality framework may therefore clarify how UBE3A biology connects molecular mechanism, developmental timing, and precision therapeutic design.

Abstract: Major depressive disorder (MDD) is clinically heterogeneous, and peripheral inflammatory biomarkers may help clarify early biological mechanisms before illness chronicity or pharmacologic treatment confound interpretation. This systematic review synthesized evidence on peripheral inflammatory biomarkers in first-episode, drug-naïve major depressive disorder (FEDN-MDD) compared with healthy controls and examined associations with clinical severity. Following PRISMA 2020, searches of PubMed/MEDLINE, Embase, PsycINFO, and Scopus from inception to March 19, 2026 identified 313 records; after screening, 16 publications were included in qualitative synthesis. Studies varied in age group, biological matrix, assay platform, and statistical reporting, precluding meta-analysis. The most frequently assessed biomarkers were IL-1β, TNF-α, IL-6, and CRP/hs-CRP. IL-6 showed the clearest recurrent tendency toward elevation in FEDN-MDD, whereas CRP/hs-CRP findings were partially positive but methodologically limited. TNF-α and IL-1β findings were mixed, and clinical correlations with depressive severity were sparse and inconsistent. Overall, the evidence supports heterogeneous early immune dysregulation in a subset of patients with FEDN-MDD rather than a single reproducible inflammatory signature. Peripheral inflammatory biomarkers should currently be considered research tools for biological stratification and mechanistic hypothesis generation, pending larger standardized longitudinal studies.

Abstract: To address the large model size, high computational cost, and limited deployment re-sources of keyword spotting models on edge platforms, this study proposes a collaborative multi-compression acceleration framework for lightweight deployment. Built on an end-to-end convolutional neural network for keyword spotting, the framework integrates adaptive structured pruning, hardware-friendly mixed-precision dynamic quantization, and quantization-aware multi-stage knowledge distillation into a unified compression pipeline. To eliminate the influence of inconsistent training budgets and data partitions across different compression branches, the results of quantization, pruning, distillation, and joint compression are reorganized under a unified evaluation protocol with mul-ti-seed mean ± std reporting. Under this protocol, the retrained baseline reaches 97.13% ± 0.85. Experimental results show that, in the quantization branch, MPDQ achieves 95.78% ± 1.69 with a compression ratio of 9.56×, demonstrating the most favorable balance be-tween accuracy and storage efficiency; in the pruning branch, AIASP reaches 95.63% ± 0.67 at 30% sparsity with a compression ratio of 1.43×, indicating a balanced compromise between accuracy retention and stability; in the distillation branch, PMKD, Multi-Teacher KD, and Fixed-T KD achieve 96.81% ± 0.69, 95.99% ± 1.18, and 96.70% ± 0.74, respectively, showing that the student model can maintain strong recognition performance under ap-proximately 4× structural compression; and the final joint compression scheme reaches 96.16% ± 0.53 with a trade-off score of 4.26 at a compression ratio of 9.89×. These results indicate that the main advantage of collaborative multi-compression lies in achieving a more balanced optimization among accuracy, model size, and compression efficiency un-der stringent deployment constraints.

Abstract: The human gut microbiota plays a central role in shaping host immunity, metabolic homeostasis and resistance to infection. Beyond microbial metabolites, increasing evidence highlights the importance of microbial and probiotic-derived proteins as key mediators of host-microbe communication. These proteins participate in immune signalling, epithelial barrier regulation and competitive interactions with intestinal pathogens. This review synthesizes current knowledge on the protein biochemistry of gut microbes and probiotics, emphasizing their mechanisms of immune modulation and roles in host-pathogen interactions. We discuss surface-associated proteins, secreted effectors, peptides and extracellular vesicle associated proteins that influence innate and adaptive immune responses. Furthermore, we explore how probiotic strains counteract pathogenic microbes through protein-mediated mechanisms and immune training. Finally, we highlight translational implications, emerging technologies and future directions for protein focussed microbiome research. This integrative perspective aims to advance the mechanistic understanding of gut microbiota-immune interactions and inform the development of next generation probiotic and therapeutic strategies.

Abstract: Background: Drug-resistant tuberculosis (DR-TB) remains a major threat to global tuberculosis control, particularly in high-burden rural settings where transmission is driven by both biological and socio-structural determinants. Although genomic surveillance and mathematical transmission modelling have improved understanding of resistance evolution and transmission dynamics, these approaches often insufficiently incorporate community-level behavioral and social drivers of disease spread. This study integrated Community-Based Participatory Research (CBPR) within a genomic–epidemiological modelling framework to develop a biosocial understanding of DR-TB transmission dynamics in a rural South African setting. Methods: Whole-genome sequencing (WGS) was performed on 32 Mycobacterium tuberculosis isolates to identify resistance-associated mutations, phylogenetic lineages, and potential transmission clusters. A deterministic two-strain transmission model distinguishing drug-sensitive (DS) and drug-resistant (DR) tuberculosis was developed to simulate transmission dynamics. CBPR-informed mechanisms were incorporated into the model through modifications of key epidemiological parameters, including transmission rate (β), treatment initiation rate (γ), and resistance amplification rate (α), reflecting the influence of community engagement, treatment adherence, and health-seeking behavior. Scenario-based simulations evaluating varying levels of community-engaged interventions were conducted over a 10-year period. Results: A substantial burden of drug resistance was observed, with 84.4% of isolates resistant to at least one anti-tuberculosis drug. Multidrug-resistant tuberculosis (MDR-TB) accounted for 46.9% of isolates, while recurrent combinations of resistance-associated mutations suggested ongoing transmission of resistant strain lineages. Lineage 2 (Beijing genotype) and Lineage 4 predominated, with advanced resistance patterns occurring mainly within Lineage 2 isolates. Model simulations indicated that CBPR-informed interventions could reduce DS-TB transmission by approximately 40–60% and DR-TB transmission by 20–35%. Scenario-based estimates also indicated a higher transmission potential for DR-TB (R₀ ≈ 2.04) than for DS-TB (R₀ ≈ 1.29). Community-engaged interventions reduced transmission by improving treatment adherence, earlier diagnosis, and enhanced infection-prevention behaviors. Conclusions: Integrating CBPR into genomic and transmission modelling frameworks provides a novel biosocial approach for understanding tuberculosis dynamics in high-burden settings. The findings suggest that community-engaged interventions can substantially influence key epidemiological drivers of transmission and resistance amplification. Embedding community participation within TB surveillance and control strategies may strengthen efforts to reduce transmission, improve treatment continuity, and address the social determinants underpinning DR-TB persistence in rural settings.

Abstract: Cardiotonic steroids (CTS) can modulate central nervous system function through their interaction with the Na⁺,K⁺-ATPase, affecting dopaminergic transmission. While the CTS ouabain is known to induce mania-like behavior and oxidative damage, the effects of other CTS are less clear. This study examined the effects of 14-day intracerebroventricular administration of 1.5 μl 100 μM marinobufagenin (MBG) on locomotion, gait, monoamine metabolism, and oxidative stress markers (MDA, SOD, catalase, MAO-B) in C57BL/6 mice. Chronic MBG caused increased locomotor activity and time spent in the center of the open field. Unlike ouabain, chronic MBG did not impair motor function, evaluated via gait analysis. MBG elevated striatal MAO-B activity and reduced prefrontal MDA levels, with no changes in SOD or catalase, indicating that it did not cause oxidative stress. However, it did affect dopamine and serotonin metabolism. Monoamine tissue content evaluation on day 15 showed increased dopamine turnover in the striatum and brain stem, and decreased it in the thalamus. Norepinephrine levels increased in the striatum and hippocampus. Serotonin turnover increased in the prefrontal cortex. These results indicate that chronic MBG increases locomotion and reduces anxiety-like behavior through region-specific modulation of dopaminergic and serotonergic signaling distinct from that caused by ouabain.

Abstract: Background: Staphylococcus argenteus is a recently recognized member of the Staphylococcus aureus complex that is almost identical to S. aureus phenotypically and by 16S rRNA gene sequences. Although genomic analyses demonstrate that S. argenteus is phylogenetically distinct from S. aureus, the two species exhibit more than 90% nucleotide identity and routine identification methods—including routine biochemical assays and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS)—cannot reliably distinguish between the two. Objectives: We develop and validate a MALDI-TOF MS–based model for accurate identification of S. argenteus. Methods: A multiplex PCR assay targeting crtM and NRPS genes served as the reference standard. MALDI-TOF MS spectra from 25 S. argenteus and 25 methicillin-susceptible S. aureus (MSSA) isolates were analyzed using ClinProTools to identify characteristic peaks and develop the identification model. The model was validated using 40 S. argenteus and 80 MSSA isolates, then applied to 130 randomly selected clinical isolates. Results: Five characteristic peaks—m/z values 5005, 5285, 5323, 6440, and 6526—were identified. Isolates exhibiting at least 4 of these 5 peaks were classified as S. argenteus; those exhibiting fewer than 4 were classified as S. aureus. The model achieved 100% specificity and 100% sensitivity in both the development and validation phases. In the clinical application phase, the model correctly classified all isolates, whereas conventional MALDI-TOF MS yielded several misidentifications. Conclusions: The identification model, and the simple peak-based rule it is based on, can accurately distinguish S. argenteus from MSSA, offering a practical diagnostic tool for clinical microbiology laboratories.

Abstract: Emerging-market equity exchanges require regime forecasting systems that are continuous in time, robust to heavy-tailed distributions, and optimised against false alarms. No existing method addresses all three simultaneously, and no prior study has reported a crisis false alarm rate on JSE equities. We propose S-NODE-ANF-RRC: a Stochastic Neural ODE embedded within an Adaptive Neuro-Fuzzy Risk-Regime Clustering architecture, motivated by the Heston stochastic volatility framework and integrated by a Milstein scheme with Lyapunov-regularised dual-loss training. The system is evaluated as a one-step-ahead probabilistic forecaster (h = 1 trading day) on 2696 daily observations across 17 JSE securities (March 2015–March 2026). Gaussian mixture clustering on raw features (kurtosis 54.8) inflates ARI by 1.3×; log-transformation corrects this systematic artefact. Two operational profiles emerge after correction: the N-ODE-ANF-RRC achieves the lowest cost (10,350 bp, 65.1% below GMM), longest lead time (0.71 days), and best MCC (0.596); the S-NODE-ANF-RRC achieves the lowest false alarm rate among probabilistic architectures (FAR = 0.051, log-loss = 1.07), with a 42.0% cost reduction versus GMM (bootstrap 95% CI [5, 250, 19, 600] bp; McNemar p = 0.027). Ablation confirms drift, diffusion, and dual-loss as the minimum viable daily-frequency configuration. The interdisciplinary fusion of physics-informed SDE dynamics, time series forecasting, and fuzzy interpretability yields two complementary JSE risk tools: an early-warning forecaster (N-ODE) and a low-false-alarm crisis classifier (S-NODE). Code and data: https://doi.org/10.5281/zenodo.19787658.

Abstract: In tropical island cities, the combined pressures of rapid high-density urbanization and year-round hot-humid climates make the pedestrian-level wind environment a critical determinant of outdoor thermal comfort and cooling performance. Focusing on Haikou, a tropical island city, this study optimizes building layouts on commercial plots under constant development intensity. A Pedestrian-level Cooling Performance Index (PLCPI) was constructed, prioritizing summer cooling and winter wind control through an AHP-EWM combined weighting method. The index integrates maximum pedestrian-level wind speeds and amplification factors to evaluate 65 layout configurations, including detached, row, perimeter, and courtyard types. The results reveal a nonlinear relationship between building count and cooling performance. Single-building layouts achieve the highest mean PLCPI (2.367), three-building layouts the lowest (1.825), prone to ventilation stagnation, and four-building layouts show a performance rebound (2.271) with stable efficiency. Crucially, spatial enclosure form is the decisive determinant under a constant building count: the optimal two-building layout B-8 (PLCPI=2.456) surpasses the best single-building layout A-2 (2.419), demonstrating that well-designed dispersed layouts can outperform centralized ones. This study proposes a dual-season adaptive evaluation framework for tropical commercial plots and reveals the nonlinear mechanism between building quantity and cooling performance, providing a scientific basis for fine-grained urban design in tropical island climates.

Abstract:

We establish new Lyapunov stability theory for ψ-Caputo fractional-order systems by strengthening Lyapunov functions under reasonable guiding wings of Class-K_∞ functions and their fractional derivative inequalities. The new generalized ψ-Gronwall inequalities and conceptual definitions of stability that are linked with the ψ-Mittag-Leffler function were introduced. Our main results are Lyapunov stability theorems whenever one finds a potential Lyapunov function that has upper and lower bounds and obeys typical Lyapunov fractional differential inequalities along imagined real trajectories of such systems. This theory works with some typical worked-out dynamic models, in which the stability dynamics are discussed.

Abstract: Pollen constitutes the primary source of proteins, amino acids, lipids, sterols, vitamins, and minerals for honey bees. However, not all pollen types provide the same resources or have the same biological value. Its chemical composition changes according to botanical origin, geographic location, and environmental conditions. This variability can influence metabolism, the immune system, oxidative balance, and the ability to resist or tolerate infections. This article examines the available evidence on the relationship between pollen chemical quality and the dynamics of Deformed Wing Virus (DWV) infection in Apis mellifera. The analysis is approached from molecular, physiological, ecological, and seasonal perspectives. Current findings suggest that more diverse and higher-quality pollen diets are generally associated with greater colony survival and improved health status, although their effects on viral load are more heterogeneous and context-dependent. In some studies, pollen intake is linked to a reduction in DWV, while in others the viral load remains stable, but bees survive longer or show better health indicators. These differences suggest that pollen may act not only by enhancing resistance to the virus but also by increasing tolerance to infection-associated damage. The potential role of pollen bioactive compounds, particularly flavonoids and phenolic acids, is also discussed. Nevertheless, evidence of direct antiviral action of these compounds in bees remains limited, as many proposed mechanisms derive from other organisms. This synthesis provides an integrative perspective on pollen nutrition and its relevance for colony resilience against viral infections.

Abstract: Transferosomes have emerged as one of the most extensively investigated ultradeformable vesicular systems for drug delivery, particularly for non-invasive administration across biological barriers. Their distinctive architecture, typically composed of phospholipids and edge activators, confers enhanced membrane flexibility compared with conventional liposomes, enabling improved adaptation to restrictive biological environments. Despite the growing body of literature, the field remains characterized by inconsistent terminology, heterogeneity in formulation strategies, and significant variability in characterization methods, which hinder meaningful comparison across studies and limit translational progress. This review provides a critical and integrated analysis of transferosomes, focusing on the relationship between formulation design, vesicle properties, deformability, and biological performance. The structural basis of transferosomes is examined with emphasis on the role of phospholipids, edge activators, and auxiliary components in modulating membrane organization, encapsulation behavior, colloidal stability, and drug release. Key quality attributes, including vesicle size, size distribution, surface charge, morphology, encapsulation efficiency, and physical stability, are discussed together with the main analytical approaches used for their evaluation. Deformability is addressed as the central functional feature of transferosomes, highlighting current experimental methods, sources of variability, and limitations affecting reproducibility and inter-study comparability. The interaction of transferosomes with biological barriers is critically examined, including the ongoing debate regarding intact vesicle penetration versus drug release prior to permeation. Major therapeutic applications are summarized, and transferosomes are positioned in comparison with conventional liposomes, ethosomes, and transethosomes within a context-dependent framework. Finally, key translational challenges are analyzed, including limited standardization, scalability constraints, storage instability, and regulatory uncertainty. In this context, this review establishes a structured framework linking formulation design, deformability, and biological performance, and identifies the critical parameters that must be controlled to enable reproducible, scalable, and clinically relevant transferosome-based drug delivery systems.

Abstract: The major challenge limiting the application of terahertz(THz) technology lies in the significant attenuation of THz waves loss of THz waves during free-space transmission arising from water vapor absorption and gas molecule scattering. Compared with free space propagation, low-loss and stable transmission of THz wave can be achieved through the waveguide. Waveguide transmission at low THz frequencies has attracted considerable attention, particularly at around 300 GHz (0.3 THz). Among the various types of THz waveguides, hollow waveguides offer a simple structure, ease of fabrication, low cost, and excellent transmission performance in the THz regime. Here, we design and fabricate a low-loss THz metal dielectric hollow waveguide based on polypropylene (PP) tubing, where an external silver film coated on the PP tube forms a leaky-type hollow waveguide structure. The linear transmission loss is measured to be 1.35 dB/m at 300 GHz. By optimizing this low-loss THz hollow waveguide, we achieve a far-field THz digital holographic (TDH) imaging recording configuration for the first time. To evaluate the imaging performance, different types of samples are measured. Experimental results for a plastic plate with aluminum strips validate a lateral resolution of ∼2.5 mm. The proposed method holds potential as a powerful tool for investigating spontaneous phenomena in the THz band.

Abstract: Bergamot-derived products have gained increasing interest as dietary supplements in small animal nutrition, due to their potential to improve meat quality, and provide functional bioactive compounds. This paper investigated the effects of feeding dried bergamot pulp on performance and meat quality in Japanese quails. 140 quails aging 15 days were divided into two groups of 70 quail each (7 replicates of 10 quails for group) and fed, for three weeks, a basal diet (control group) or the basal diet in which part of the maize was replaced with 10% of dried bergamot pulp (BP10 group). The integration of dried bergamot pulp (BP) reduced dry matter i ntake, a verage d aily g ain, a nd consequently final body weight compared to control treatment. Quails from BP10 group showed the highest feed conversion ratio. The BP10 treatment tended to increase eicosapentaenoic acid and ω-3 PUFA. The inclusion of bergamot pulp in quail diet did not alter the TBARS value in meat. Color analysis showed that the integration of bergamot into the quail diet led to higher lightness (L*) and yellow index (b*) values compared to the control group. This study demonstrates that dietary supplementation with 10% of BP to quails led to a reduction in dry matter i ntake ( DMI) a nd consequently in average daily gain (ADG) in quails. However, a positive trend toward an increase in ω-3 fatty acids has been observed.

Abstract: Background/Objectives: Probiotic interventions are widely used to improve intestinal health; however, comparative evidence on multi-strain formulations with different po-tencies, particularly when combined with plant-based complexes, remains limited. eval-uated the effects of two probiotic blends containing phytonutrients (PBP1 and PBP2) on bowel function, microbial metabolites, and gut barrier-related markers. Methods: In this randomized, double-blind, placebo-controlled trial, participants received PBP1, PBP2, or placebo for 8 weeks. Stool patterns (7-day Bristol Stool Form Scale (BSFS) diary), fecal short-chain fatty acids (SCFAs), tryptophan metabolites, zonulin, and gut microbiota were assessed at baseline and Week 8. Efficacy was evaluated by comparing each inter-vention group with the placebo group. Results: Both PBP1 and PBP2 significantly in-creased the proportion of normal stool types (BSFS types 3–5) compared with placebo (p < 0.05). Fecal SCFA levels, including acetate, propionate, and butyrate, were significantly increased in both intervention groups. Notably, butyrate levels were significantly ele-vated compared with placebo. Fecal tryptophan levels decreased, while indole metabo-lites showed increasing trends, with an inverse correlation observed between tryptophan and indole, particularly in the PBP2 group. Fecal zonulin showed a decreasing trend, with significant reductions in participants with 25.0≤BMI<30.0 kg/m². Microbiome analysis revealed preserved alpha diversity with selective compositional shifts, including en-richment of Lactobacillus-related taxa. Conclusions: Supplementation with PBP1 and PBP2 improved bowel function and was associated with changes in microbiome-derived metabolites, including SCFAs and tryptophan–indole metabolism, with BMI-dependent changes in barrier markers. These findings suggest a potential role of microbi-ome-mediated metabolic modulation in intestinal health.

Abstract: Human fertility has declined sharply since 1950, and a growing body of evidence suggests that conventional socioeconomic explanations do not fully account for the timing and breadth of this trend. This review examines the Circadian-Light Hygiene hypothesis, which proposes that daily light exposure is a fundamental regulator of reproductive health. We synthesize findings from photobiology, endocrinology, reproductive medicine, and epidemiology to evaluate how artificial light at night, insufficient daytime light, and irregular light-dark patterns may disrupt the hormonal timing systems that support reproduction. The available evidence indicates that such disruption can alter melatonin signaling, circadian gene regulation, and neuroendocrine rhythms, with downstream effects on ovulation, sperm quality, endometriosis, polycystic ovary syndrome, pregnancy outcomes, and developmental programming. Urbanization, screen use, and shift work appear to amplify these effects, while genetic variation may modify individual susceptibility. Although direct causal evidence in humans remains limited for several endpoints, the convergence of observational, experimental, and translational data supports circadian light misalignment as a plausible and potentially modifiable contributor to fertility decline. Optimizing daily light exposure may therefore represent a low-cost and scalable strategy for improving reproductive health.

Abstract: Construction companies, petrochemical companies, and airports are classified as large-scale organizational and technical systems. In organizational and technical system is implemented using hierarchical distributed control systems. To achieve goals, numerous parallel technological operations and business processes are executed, requiring synchronization and consuming significant energy resources. To optimize energy resources at all hierarchical levels, an up-to-date picture of the system state is necessary. This paper proposes constructing a snapshot of the system's state, which allows for an assessment of system performance within selected criteria and the selection of appropriate solutions within the entire control system. Procedures for constructing a snapshot are discussed. An example of optimizing the energy consumption of a control system based on snapshot analysis is provided.

Abstract: This study aims to evaluate multiple feature sets composed of sensor-based biomarkers acquired during walking for the automated estimation of post-stroke motor impairment levels using Fugl-Meyer Lower Extremity Assessment (FMA-LE) derived classes. Sensor-based walking data from the open-source ARRA dataset were combined with data collected at the Hospital of Braga. Data from 32 post-stroke individuals (FMA-LE:24±3) were included. A decision tree classifier was evaluated using stratified 6-fold cross-validation across different feature configurations, including: correlated versus full feature sets; spatiotemporal versus electromyographic (EMG) features; inclusion of demographic variables; and the use of data augmentation. The best performance was achieved using correlated EMG features combined with age, paretic side, and body mass, along with noise-based data augmentation, yielding a validation MCC of 0.85±0.16 and a test MCC of 0.70. EMG features provided improved classification performance compared to spatiotemporal features, and comparable results were obtained using a reduced subset of muscles. These results demonstrate the feasibility of using EMG-based features acquired during walking to classify post-stroke motor impairment levels. Feature reduction and inclusion of demographic variables may support efficient model design, while data augmentation may enhance generalization. Further validation in larger and more diverse datasets is required to assess robustness and clinical applicability.

Abstract: Background/Objectives: African Swine Fever (ASF) represents one of the most serious threats to animal health and global food security. The causative agent of ASF is the African swine fever virus (ASFV), a DNA virus belonging to the Asfarviridae family. Here, we describe ex vivo results for an original anti-ASFV vaccine approach based on the cellular immune response induced by extracellular vesicles (EVs) engineered to express four ASFV antigens. EV engineering was achieved by expressing a DNA vector encoding a biologically inactive HIV-1 Nef protein (Nef^mut), which exhibits unusually high efficiency of incorporation into EVs, even when fused to foreign proteins. Previous studies have demonstrated that intramuscular injection of Nef^mut-based vectors leads to the engineering of EVs spontaneously released by muscle cells and induction of antigen-specific CD8⁺ T cell immunity. Methods: We designed DNA vectors expressing the fusion products between Nef^mut and each of the four ASFV structural proteins p³⁰, p⁵⁴, pp⁶², and p⁷². Engineered EVs were molecularly characterized by western blot and nanotrack analysis, and their potential immunogenicity was assessed by priming and cross-presentation assays. Results: We assessed that the four antigens were successfully expressed in transfected mammalian cells, with the release of valuable amounts of engineered EVs. When immature swine dendritic cells were challenged with the engineered EVs and then co-cultivated with autologous peripheral blood lymphocytes in priming assays, lymphocyte subpopulations specifically reacting against each ASFV antigen were elicited, as detected by an IFN-γ ELISpot assay. In addition, we provide evidence that the Nef^mut-based fusion products incorporated into the engineered EVs can be cross-presented by professional antigen-presenting cells, leading to cross-priming of autologous lymphocytes. Conclusions: These results represent the best premise to go forward with experiments of immunogenicity and antiviral efficiency in pigs.