Abstract: The present study examined the effect of Enterococcus durans cell free supernatant (CFS) on interleukin (IL) 8, 10 and 1β gene expressions in the intestinal cell line HT-29 treated with Staphylococcus aureus CFS. HT-29 cells were incubated with E. durans CFS or S. aureus CFS, or S. aureus CFS plus E. durans CFS. All concentrations of E. durans CFS did not show cytotoxicity, while the highest treatment (44.9 μg/mL) with S. aureus CFS induced significant cell death. S. aureus CFS did not modify IL-1β gene expression, while E. durans CFS alone or in combination with S. aureus CFS reduced it. Treatment with S. aureus CFS induced greater expression of the IL-8 gene compared to S. aureus CFS plus E. durans CFS. S. aureus CFS alone or in combination with E. durans CFS increased the expression of the IL-10 gene, while E. durans CFS alone did not modify it. These results suggest a potential protective role of the E. durans secretome in mitigating the inflammatory environment in intestinal cells. This treatment could be useful to protect against possible contact with dangerous soluble microbial products present in food.

Abstract: Salmonella enterica remains a major threat to animal and human health because of its broad host range, increasing antimicrobial resistance (AMR), and capacity to form biofilms. Biofilm formation enhances bacterial persistence in host tissues, farm environments, food-processing systems, and clinical reservoirs, while also contributing to their tolerance against antibiotics, disinfectants, and other stresses. However, biofilm capacity is not uniform across serovars and is influenced by host adaptation, niche specialization, and accessory genome content. This review synthesizes current knowledge on the relationship between biofilm formation, AMR, and serovar-specific adaptation in Salmonella. It examines biofilm-associated traits across various hosts (e.g., gastrointestinal tract and gallbladder, and environmental (e.g., food-production and clinical) niches, and discusses comparative evidence from genomic, transcriptomic, proteomic, and metabolomic studies. Particular attention is given to the emerging concept of comparative biofilmomics, which integrates phenotypic and multi-omics data across diverse serovars and host sources to identify conserved and niche-specific determinants of persistence. This framework may help define high-risk lineages that couple multidrug resistance (MDR) with enhanced biofilm-forming capacity. A better understanding of these linked traits will support the development of more targeted interventions for controlling persistent Salmonella in veterinary, food production, and public health settings.

Abstract: Bedding serves a vital role in horse stable management. Although earlier research has examined stress responses to bedding materials, the effect of bedding types on these responses in horses of different sizes has not been studied. This study assessed the influence of bedding materials (straw vs rice husk) and body size (horses taller than the upper bar of the front gate [H1] vs shorter ones [H2]) on stress responses. Stress was evaluated using behavioural scores, recumbent time, and autonomic regulation via heart rate (HR) and heart rate variability (HRV). Microenvironmental variables such as relative humidity (RH), air temperature (AT), volatile organic compounds (VOCs), carbon dioxide (CO₂), particle counts (PC; PC1, 2.5 and 5) and fine particulate matter (PM; PM1, 2.5 and 5) were also recorded. No differences were observed in RH, AT, VOCs or CO₂ between stables with different bedding materials. However, PC (1, 2.5 and 5) and PM (1, 2.5 and 5) levels were higher in stables with rice husk than in those with straw. Beat-to-beat (RR) intervals increased, and HR decreased in both H1 and H2 horses on straw during the night (p < 0.05). H1 horses generally showed lower HR and higher RR intervals than H2 horses during housing, regardless of the bedding material used (p < 0.05–0.0001). Although bedding and body size affected behavioural scores and several HRV metrics, no significant within- or between-group differences were detected. These findings suggest that both bedding material and body size influence stress responses; however, short-term individual box housing may not provide sufficient stressors to produce significant differences.

Abstract: The enteric nervous system (ENS) can function independently of the central nervous system (CNS) in regulating complex gastrointestinal processes and may possess forms of learning and memory. Recently, we have proposed that maternal stress, depression, or anxiety during pregnancy may induce stress-related long-term epigenetic implicit memory (SLEIM) in the foetal ENS via mechanisms distinct from those of the CNS. These stress imprints may persist throughout life. Through the bidirectional microbiota–gut–brain axis (MGBA), SLEIM signals originating in the embryonic ENS may influence the CNS. Because these signals are implicit and unrelated to conscious representation, the CNS cannot directly interpret them, yet it must regulate their physiological consequences. This interaction may activate stress-response systems, including the hypothalamic–pituitary–adrenal (HPA) axis and immune pathways, leading to cortisol release, mast cell activation, and cytokine imbalance. A self-reinforcing cycle may thus develop between the ENS and CNS. The frequent comorbidity of fibromyalgia (FM) and irritable bowel syndrome (IBS) suggests shared pathogenic mechanisms, particularly central sensitization and MGBA dysfunction. Clinically, patients with FM often display childlike behavioural traits. We hypothesize that early developmental influences—potentially linked to maternal SLEIM effects on the foetal ENS—may contribute to personality immaturity in FM, similarly to IBS. Under varying biopsychosocial conditions, IBS-related mechanisms may later manifest as systemic symptoms characteristic of FM.

Abstract: Chronic alcohol consumption increases the risk of osteoporosis and fracture by disrupting bone remodeling, in part by enhancing osteoclastogenesis. However, the cellular mechanisms underlying this process remain incompletely defined. We analyzed scRNA-seq data from osteoclasts differentiated in vitro from bone marrow mononuclear cells obtained from macaques following 12 months of chronic ethanol or isocaloric control solution consumption. Module scoring, trajectory inference with generalized additive modeling (tradeSeq), and CellChat-based analyses of intercellular communication were applied to uncover ethanol-induced changes in metabolic reprogramming, lineage progression, and signaling network dynamics. Module scoring indicated metabolic reprogramming toward oxidative phosphorylation, with reduced glycolytic, migratory, and phagocytic activities. Pseudotime analysis revealed accelerated osteoclast lineage commitment, broader intermediate differentiation states, and stabilization of mature osteoclasts. CellChat analysis showed globally amplified intercellular signaling, with mature osteoclasts functioning as dominant communication hubs sustained by autocrine feedback. Together, chronic alcohol consumption rewired osteoclastogenesis through early fate priming, metabolic adaptation, and hierarchical remodeling of intercellular communication, promoting enhanced osteoclastogenesis. These findings provide mechanistic insight into alcohol-induced bone pathology and highlight potential targets for therapeutic intervention.

Abstract: Is an ant colony conscious? What about a group of people talking, a cloud-hosted language model, or even a galaxy? Can a conscious mind only get so big? Does consciousness depend only on what is computed, or when and where? I see two possibilities affecting the answers to these questions. I name them Chord and Arpeggio, and formalize the distinction mathematically. If the ingredients of a subjective experience must be simultaneously true at one objective instant and causally exchange influence within a time window θ, then the system diameter D satisfies D ≤ κvθ, where v is the signal speed ceiling and κ depends on exchange architecture. I call this requirement Chord, because it is like a musical chord whose notes sound together. The alternative is Arpeggio. It asks only that each ingredient occur somewhere in the window. I prove that Arpeggio is strictly weaker than Chord, and that architectures with limited concurrency can satisfy Arpeggio while structurally forbidding Chord. I argue for Chord on formal, neuroscientific, and architectural grounds. A mechanistic model confirms a fragmentation transition at the theoretical threshold. I examine primate corpus callosum data to estimate empirical lower bounds on θ. I provide case studies showing that under Chord, ant colonies and human populations are ruled out as single conscious entities, cloud-hosted AI is constrained by co-instantiation rather than diameter, and brain-computer interface hybrids face latency-dependent limits. A mind can only get so big. Arpeggio is far more permissive, implying consciousness seemingly everywhere.

Abstract:

Background/Objectives: The escalating crisis of antibiotic resistance and the inherent limitations of conventional antibiotics necessitate the development of innovative therapeutic strategies. Targeted drug delivery (TDD) offers a powerful approach to enhance efficacy, minimize systemic toxicity, and circumvent bacterial resistance. This systematic review aims to evaluate the potential of unique bacterial transport systems (BTSs) and surface specific receptors as platforms for TDD via the "Trojan Horse" strategy (THS). Methods: A comprehensive literature review was conducted, focusing on studies that investigated the specificity and mechanisms of BTSs responsible for the uptake of essential metabolites. This includes an analysis of transport systems for siderophores, bacteria-specific sugars, cell wall components, D-amino acids, and vitamins. We assessed preclinical and clinical examples of drug conjugates utilizing these pathways, as well as emerging platforms such as bacteriophage-derived proteins, antibody-antibiotic conjugates, and bacterial extracellular vesicles (EVs). Results: BTSs demonstrate high specificity for their cognate substrates, providing effective molecular gateways for drug conjugate import. The siderophore-cephalosporin conjugate cefiderocol represents a clinically validated example, having received FDA approval. Preclinical studies further reveal that conjugates utilizing sugars (e.g., maltose, trehalose) and vitamins (e.g., B12) can significantly enhance antibiotic uptake and activity against both Gram-positive and Gram-negative pathogens, including drug-resistant strains. Emerging platforms like bacteriophage endolysins and engineered EVs show promise for overcoming biological barriers such as bacterial outer membranes and intracellular host niches. Conclusions: The THS leveraging BTSs represents a clinically viable and promising avenue for next-generation antibacterial therapies. While significant progress has been made, including regulatory approval of cefiderocol, further research is critically needed to identify novel BTSs, optimize drug-linker chemistry, improve the pharmacokinetics and biosafety of conjugates, and translate these innovative platforms into effective treatments for drug-resistant infections.

Abstract: Raman Spectroscopy is non-destructive, label free analytical technique that can probe the biochemical alterations in tissues and cell. Raman Spectroscopy, being sensitive to biochemical perturbations, can potentially provide early and real-time identification of changes proceeding morphological changes, allowing early diagnosis as well as diseases monitoring. Recent research has demonstrated its broad utility across diverse clinical domains, including cancers, neurological conditions and infections. Raman spectroscopy combined with machine learning algorithms allows rapid assessment and automated pipelines and can act as a clinical adjunct, enhanced by integrating tools like principal component analysis (PCA), linear discriminant analysis (LDA), random forests, and deep learning architectures. These models allow interpretation of complex spectra, and decipher meaningful biomarkers in heterogeneous clinical samples. This review highlights the earliest and recent progress in Raman based non-destructive diagnosis underscoring advances in cancer diagnosis and challenges faced in clinical settings.

Abstract: Hydroponic fodder production can be applied to a wide range of forage crops, optimizing the use of water, arable land, and other resources in a sustainable manner. This study aimed to evaluate the effects of hydroponically sprouted seeds on bull meat quality. Wheat, barley, alfalfa, and peas were germinated in a hydroponic system for seven days, and dairy–meat crossbred bulls were fed these sprouts for 18 months. The chemical composition of the sprouted seeds, including proteins, fats, fiber, mineral elements, and amino and fatty acids, was analyzed. Samples of the Longissimus dorsi (LD) muscle were collected from the carcasses for comprehensive meat quality assessment. The study revealed that while sprouted grains did not significantly alter the chemical composition of meat, they influenced its physical characteristics, depending on the type of grain. Meat from bulls fed sprouted alfalfa seeds was significantly lighter in color and contained less water. The inclusion of sprouted grains also positively affected the amino acid and fatty acid profiles. The content of undesirable saturated fatty acids was 13.78–15.70% lower compared with meat from the control group. Furthermore, meat from bulls receiving sprouted alfalfa seeds contained 2.86% more essential amino acids, while meat from bulls fed sprouted peas contained 0.58% more than the control. Overall, the addition of sprouted grains enhanced the biological value of beef protein, with alfalfa seeds demonstrating the greatest effect on protein integrity indicators.

Abstract:

Jatropha curcas L. is a bioenergy crop of interest because of the high oil content in its seeds suitable for conversion into biofuels. However, its oil content is extremely variable among accessions and the mechanism of oil accumulation is poorly understood in this oleaginous species. In this study we analyzed cloned plants of three chemotypes of J. curcas collected in Chiapas, Mexico: CAC-3, COM-1 and MAP-2. All are monoecious and accumulating different amounts of seed oil: 10 %, 30 % and 54 %. We studied the expression of the β-ketoacyl-acyl carrier protein (ACP) synthase genes (KAS) in developing seeds and their relationship with the content and composition of the oil. Differences of the levels of expression of the KASI and KASII genes were found, while KASIII gene was expressed at high levels in all three chemotypes. The expression of KASI and KASII was statistically associated to the oil accumulation. Results of this study are discussed based on the regulation of the transcription of the KAS genes, in order to contribute to the understanding of the oil accumulation in the seed and could be of value for designing biotechnological strategies with which to improve this species.

Abstract: Astaxanthin (AST), despite its high bioactivity, exhibits poor stability and low bioavailability due to its strong lipophilicity and inherent degradation susceptibility. To overcome such a challenge, we developed a food-grade oleogel delivery system using a soy protein-arabinoxylan (SA) glycosylated complex modulated by different concentrations (0.5%–3%) of sucrose ester (SE) or soy lecithin. We show that the emulsifier concentration has a non-linear effect on the oleogel microstructure: an optimal level of 1% had a significant impact on the interfacial compactness and network density, giving rise to improved thermal stability, rheological strength and AST encapsulation efficiency (81.27%). During in vitro digestion, the SA matrix in combination with emulsifiers allowed gastric protection and intestinal-targeted release of AST with a bioaccessibility of up to 88.84% (SAO-SE-AST). This controlled-release profile directly translated into enhanced in vivo antioxidant efficacy in wild-type Bristol N2 Caenorhabditis elegans as evidenced by reduced lipofuscin accumulation, elevated thermotolerance (survival rate: 64.44%-73.33%), suppressed reactive oxygen species levels and activation of endogenous antioxidant enzymes (superoxide dismutase as well as glutathione peroxidase). Collectively, this research has uncovered that food-grade emulsifiers are not only stabilizers, but also key regulators of oleogel architecture and bioactive functionality. These results provide a structure-digestion-bioactivity correlation for protein-polysaccharide oleogels, representing a rational design strategy for high-performance delivery systems of lipid-soluble nutraceuticals.

Abstract:

The ‘Tenuifoliae irises’ are a distinctive group of beardless, rhizomatous perennial irises, which are characterised by their somewhat vertical rhizomes, typically clothed at the apex with long maroon-brown, sharp fibrous remains of leaf sheaths; perianth tube long, filiform to scapiform; stigma bilobed; capsules often trigonous to six-ribbed, apically beaked; and seeds angulose to subcubic or pyriform, lacking fleshy appendages, and with testa hard, irregularly wrinkled. The representatives of the aggregate are mostly native to the dry steppes and grasslands from lowland to high mountain habitats of Central and Eastern Asia, extending westwards to the Black Sea and Caspian regions. Morphological classification of the ‘Tenuifoliae irises’ recognises about ten to eleven species, which are arranged into two genera, Sclerosiphon to Cryptobasis. Diverse molecular research recovered members of the ‘Tenuifoliae irises’ in contrasting placements within the ‘Iris-flower clade’. Sometimes, Sclerosiphon was sister to Eremiris, but Cryptobasis aligned with the ‘Spuria irises’ (Chamaeiris) and the ‘Spanish irises’ (Xiphion and related genera); in other cases, both Sclerosiphon and Cryptobasis formed a clade sister to Chamaeiris, or Cryptobasis alone was identified as the basal member of the Iris s.l. clade, positioned immediately after Siphonostylis. To examine these taxonomic discrepancies within a rigorous molecular‑systematic framework and using 12 reliably authenticated specimens, we generated 24 sequences of the matK gene (12) and the trnL (UAA)–trnF (GAA) loci (12) from members of the ‘Tenuifoliae irises’. These sequences were subsequently incorporated into a comprehensive dataset of the ‘Iris‑flower clade’, enabling a broader analytical assessment. The obtained three-taxon statement hierarchy of patterns and maximum likelihood phylogenetic trees both recover the ‘Tenuifoliae irises’ as monophyletic and sister to Chamaeiris, and in turn to the ‘Xiphion s.l. clade’. We also found Sclerosiphon and Cryptobasis as sister genera. The morphological and karyological data supporting those relationships are discussed, which allow getting back to Rodionenko’s sources and recovering Sclerosiphon in his original sense, alongside Cryptobasis. Furthermore, the molecular results allow us expanding Sclerosiphon to include the Eastern Chinese members of the aggregate. In consequence, five new combinations (one series and four species) are established in the genus, one lectotype is designated, and data on nomenclature, distribution and ecology of the accepted species are reported.

Abstract: Cabbage is considered a healthy vegetable due to its chemical composition and high nutritional value. This is due to the presence of carbohydrates and dietary fiber as the main constituents, as well as the presence of vitamin C. The end product thus obtained (sauerkraut) is a low-calorie product with a long shelf-life. The most important role in the fermentation of cabbage is played by lactic acid bacteria whose activity is influenced by physical factors such as temperature and some chemical factors such as salt concentration or the addition of spices which, in addition to their flavoring effect, may also have an inhibitory effect on undesirable microflora. The present study investigates the effect of essential oils extracted from plants on lactic acid bacteria responsible for the fermentation of cabbage. Essential oils from thyme, dill, wild thyme, bay and basil were tested. The obtained results have shown that the essential oils that were added to the fermentation mass in concentrations of 0.015% did not inhibit the activity of lactic acid bacteria responsible for lactic fermentation.

Abstract: Background Generative AI (genAI) has the capacity to create realistic and convincing animal videos, however, it must simplify and reduce behavioural variation to do so, possibly leading to misinformation. Methods We categorised 29 videos in the press release for a specific video genAI engine. Twelve featured animals. We mapped each video to the Five Domains and categorised behaviour and welfare within. Results Negative welfare was rarely seen, ranging from 8% (n = 1) for Nutrition, to 42% (n =5) for Behavioural Interactions. By contrast, Mental State, Environment, and Behavioural Interactions appeared positive in >42% (n = 5) of the videos featured. However, videos were often misleading or did not represent accurate animal behaviour. Limitations This work was limited to a press-release of data and does not explore user experience. Conclusions GenAI videos pose a new route for client confusion and veterinarians need to incorporate genAI misinformation combatting in their practice.

Abstract: While falling costs have increased access to genomic sequencing, the impact of clinical sequencing is often hindered by the challenge of interpreting complex genetic data. The high prevalence of variants of unknown significance (VUSs) can lead to false reassurance or psychological distress, as patients and non-expert clinicians may misinterpret inconclusive results. We propose that artificial intelligence (AI) may serve as a critical clinical decision-support tool to improve the efficiency of genetic testing, especially in variant analysis. We advocate integrating AI throughout the genetic diagnostic workflow and outline current approaches to AI-assisted variant analysis to enable efficient personalized treatment. We also discuss anticipated challenges in this pursuit and offer recommendations to ensure precision, accuracy, reproducibility, and transparency.

Abstract: Black pepper (Piper nigrum L.) faces challenges related to irregular flowering, which compromises crop productivity. Gibberellic acid (GA₃) is a plant growth regulator known for its role in inducing reproductive processes, although its effects on this species are not yet fully understood. This study aimed to evaluate the influence of different GA₃ doses on flowering and vegetative growth in black pepper plants. The experiment was conducted with black pepper seedlings of the Bragantina cultivar in a randomized block design, with four doses of GA₃ (0, 10, 20, and 30 mg L⁻¹) and six replications, using eight-month-old plants grown in pots under full sun. GA₃ applications were performed in two floral induction cycles. Variables related to flowering, chlorophyll a fluorescence, vegetative growth, biomass allocation, and carbohydrate distribution were evaluated. The data were subjected to analysis of variance, regression, mean grouping test, and principal component analysis. The results showed that inter-mediate doses (10 and 20 mg L⁻¹) significantly stimulated flowering at early developmental stages, whereas the 30 mg L⁻¹ dose enhanced vegetative growth while reducing floral induction. Additionally, GA₃ affected physiological parameters by increasing photosynthetic efficiency and altering carbohydrate balance, with higher accumulation of soluble sugars in leaves and reduced starch content in roots. It is concluded that GA₃ application is a promising strategy to modulate reproductive transition in black pepper, with 10 to 20 mg L⁻¹ doses recommended to promote flowering without compromising plant development.

Abstract: Milk production in dairy cattle is increasingly challenged by thermal variability. This underscores the need for reliable assessment of microclimatic conditions and their interaction with animal- and management-related factors to ensure sustainable dairy production. The aim of this study was to evaluate the effects of thermal variability and selected environmental and biological factors on key milk production traits in dairy cattle. The influence of fixed factors related to production conditions and microclimatic variability, including the Temperature–Humidity Index (THI), on daily milk yield (MY), milk fat content (MF), and milk protein content (MP) was assessed. The study used a dataset covering two observation periods of daily milk production traits in cows of different breeds (Simmental, Holstein-Friesian, Red Holstein, and Brown Swiss) reared in three regions of the Republic of Serbia (Mačva, Podunavlje, and Šumadija), enabling an assessment of thermal variability under diverse production and microclimatic conditions. The expression and variability of the investigated traits were determined using the PROC FREQ and PROC MEANS procedures, while the effects of individual factors were analysed using general linear and regression models, with results expressed as least squares means. All examined factors showed a highly significant effect on MY, MF, and MP (p < 0.0001). Although the overall level of heat stress was moderate, milk production was highest within the THI range of 51–60. These findings demonstrate that thermal variability significantly influences milk production and composition and highlight the importance of integrating microclimatic indicators into sustainability-oriented dairy management and breeding strategies.

Abstract: Since the pioneering work of Dean P. M. and Matthew E. K. (1970), four decades have elapsed without any consensus on the mechanism responsible for the oscillations of the plasma membrane voltage exhibited by pancreatic β-cells stimulated by glucose. In this review, the different hypothesis dealing with the cause of voltage oscillations that lead to insulin secretion pulsatility will be commented. The earliest explanation attributed the voltage oscillations (bursting) to glycolytic oscillations, taking as a reference skeletal muscle glycolysis oscillations. Later, the scientific interest moved to glucose oxidation after discovering that some mitochondrial parameters also oscillated in synchrony with membrane voltage oscillations. As [Ca2+]cyt increases resultant from membrane depolarization oscillated in synchrony with membrane bursting, it competed with metabolic oscillations (e.g. cytosolic ATP/ADP) for being the cause or the effect of insulin pulsatility; it was demonstrated that metabolic oscillations preceded [Ca2+]cyt oscillations. We are contributing with the hypothesis attributing the cause of voltage oscillations to a sequential competition of two β-cell plasma membrane channels: K+ATP channel and Cx36 hemichannel (Cx36H). Whereas increased glucose metabolism (increased ATP/ADP) closures K+ATP channels and depolarizes the plasma membrane (active phase of a bursting), Cx36Hs are opened and repolarize the membrane potential with a certain delay by inhibiting glucose metabolism (silent phase of a bursting). Repolarization, in turn, closes Cx36H and allows the recovery of glucose oxidation and beginning of a new active phase.

Abstract: Aquaculture plays a strategic role in food security and rural development in coastal regions. However, structural, economic, and institutional constraints affect small-scale producers in heterogeneous ways. This study analyzes how small-scale aquaculture producers in Manabí (Ecuador) perceive the main challenges affecting their activity, based on a typology comprising three production systems: Backyard, Transitional, and Commercial. A structured questionnaire was administered to 98 producers, including 20 variables assessed using a five-point Likert scale. The analysis combined non-parametric univariate tests (Kruskal–Wallis with Dunn post-hoc comparisons) and multivariate techniques to identify statistically significant differences and structured perception patterns across production systems. Significant differences were detected in variables related to biological input supply, market conditions, and structural production constraints. In particular, larvae and fingerling supply, selling prices, buyer availability, and pond surface area showed differentiated perception patterns across systems. Most differences occurred between Backyard farms and the other two production systems, while Transitional and Commercial farms displayed more similar perception profiles. Transversal constraints shared across systems included high feed costs, energy expenditure, and regulatory requirements. Principal Component Analysis identified two main perception gradients related to market and input constraints and to structural and managerial limitations. Discriminant analysis further confirmed the ability of these dimensions to differentiate production systems. These findings highlight the multidimensional nature of constraints affecting small-scale aquaculture and suggest that production systems are better interpreted as gradients of pressures rather than strictly discrete categories. The results underline the need for adaptive governance approaches combining transversal measures with system-specific interventions. Overall, the study provides empirical evidence to support the design of differentiated and context-sensitive policies aimed at strengthening the sustainable development of small-scale aquaculture in Manabí and similar territories.

Abstract: Transarterial chemoembolization (TACE) is the standard treatment for patients with intermediate-stage hepatocellular carcinoma (HCC), yet nearly half of treated patients fail to achieve durable benefit, and reliable biomarkers enabling early therapeutic stratification are still lacking. Treatment response is typically assessed by imaging one month after TACE and at three-month intervals, potentially delaying timely access to alternative therapies in non-responding patients. Circulating microRNAs (miRNAs) represent promising biomarkers due to their stability in body fluids and ease of detection. Here, we evaluated circulating miR-22 as an early predictor of TACE response and as a mechanistically relevant therapeutic target. Circulating miR-22 levels were measured by microarray and quantitative RT–PCR in three independent cohorts of early-to-intermediate stage HCC patients undergoing TACE. Circulating miR-22 increased significantly in non-responders as early as 48 h after treatment, and fold changes consistently predicted treatment failure across two independent validation cohorts. Mechanistically, we identified the G2/M checkpoint kinase WEE1 as a direct functional target of miR-22. Modulation of the miR-22/WEE1 axis affected cell-cycle progression, proliferation, apoptosis, and DNA damage response in HCC cell lines and xenograft models. Under hypoxia-mimicking conditions combined with doxorubicin exposure, pharmacological inhibition of WEE1 induced mitotic catastrophe in highly proliferative miR-22–silenced cells. Collectively, these findings identify early post-TACE elevation of circulating miR-22 as a biomarker of non-response and highlight the miR-22/WEE1 axis as a potential target for precision treatment strategies in HCC.