Search Results (5,115)

High-altitude hypoxia is associated with sleep disruption, affective fluctuation, and changes in cognitive performance, yet prior findings remain markedly heterogeneous in both direction and magnitude. To clarify this variability, this structured narrative review synthesizes evidence from systematic reviews, meta-analyses, and key empirical studies on psychological and cognitive adaptation to high-altitude hypoxia, with a particular focus on personality and related traits. The review is organized along three axes: exposure context and characterization (real-altitude field/residential exposure, hypobaric chambers, normobaric hypoxia, expedition settings, and dose alignment), exposure duration and time course, and outcome domains spanning sleep/fatigue, affective adaptation, and cognitive performance. Across the literature, cognitive effects appear domain-specific and strongly conditioned by exposure dose, temporal stage, and task characteristics, whereas sleep disturbance during the early acclimatization period emerges as one of the more consistent findings and appears closely intertwined with anxiety-related responses. Although direct personality-related evidence remains limited and uneven, the available findings can be organized into a testable trait–process–outcome framework that distinguishes candidate associations with relatively more direct evidence from mechanism-informed hypotheses. Neuroticism/emotional stability and anxiety-related traits have received comparatively more direct empirical investigation than other personality domains, but the available evidence is insufficient to regard them as established predictors of high-altitude adaptation. The roles of conscientiousness, resilience/hardiness, extraversion, agreeableness, and openness remain more tentative and are better treated as targets for prospective testing. We therefore propose that personality-related dispositions may influence adaptation-relevant outcomes through candidate processes involving threat appraisal, coping, resource regulation, and social interaction. We conclude that future research should strengthen exposure dose characterization, specify the boundary conditions for generalizing across real-altitude, hypobaric-chamber, normobaric hypoxia, and expedition settings, and adopt longitudinal, multilevel, and ecologically valid designs to test personality-related pathways more rigorously. Full article

Background: Manure-amended agricultural soil is a critical reservoir of antibiotic resistance genes (ARGs), posing escalating threats to environmental health and food safety. However, the temporal trajectories of ARG prevalence throughout the complete reproductive cycle of cash crops, and their mechanistic linkages with fertilization regimes and microbial community succession, remain inadequately understood. Methods: To bridge this knowledge gap, we conducted an in situ field experiment over the entire growth period of Chinese cabbage at a long-term manure-amended farm in Tianjin, China. Six contrasting fertilization strategies were evaluated: unfertilized control (CK1), unfertilized baseline control (CK2), traditional full-rate combined manure–chemical fertilization (TF), traditional half-rate combined manure–chemical fertilization (T1), half-dose sole manure fertilizer (T2), and half-dose sole chemical fertilizer only (T3). Results: Our results demonstrated that ARG abundance and associated mobile genetic elements (MGEs) exhibited a pronounced transient surge immediately post-fertilization, yet reverted to baseline levels by harvest, revealing a tangible resilience of the soil resistome. Notably, the optimized half-organic fertilization (T2) effectively curtailed the proliferation of manure-derived pathogenic taxa while preserving beneficial keystone phyla (e.g., Acidobacteria and Proteobacteria), indicating a trade-off between nutrient provisioning and ecological filtering. Co-occurrence network analysis further identified MB-A2-108, Saccharimonadales, and Rokubacteriales as pivotal hosts for multidrug-resistant ARGs, underscoring that microbial interspecific interactions—rather than taxonomic richness alone—are the primary drivers of resistome succession. Quantitative risk assessment confirmed that the T2 regimen reduced the composite ARG contamination index (CF_zone) by 25% relative to conventional full fertilization (TF), while maintaining comparable cabbage yields. Conclusions: Collectively, our findings advocate for precision organic fertilization as a nature-based solution that synchronizes nutrient supply with crop demand, curtails ARG propagation, and mitigates long-term agroecological risks. Full article

Background: Patient-specific targeted radionuclide therapy (TRT) requires consideration not only of administered activity but also of the spatial distribution of radiopharmaceutical uptake and radionuclide-specific energy deposition. This study developed a voxel-based computational workplan to compare ¹⁷⁷Lu, ¹⁶¹Tb and ⁹⁰Y, together with hybrid radionuclide models, using patient-specific PSMA PET-derived tumour activity distributions. Methods: PSMA PET/CT data from 20 patients with prostate cancer, comprising 10 ¹⁸F-PSMA and 10 ⁶⁸Ga-PSMA examinations, were processed to obtain 2285 quality-filtered lesions. Radionuclide-specific dose-point kernels (DPKs) were generated in water using OpenGATE and applied to voxel-wise lesion activity distributions to reconstruct absorbed-dose maps. Kernel characteristics were evaluated using radial energy-containment metrics, and ¹⁷⁷Lu, representing ¹⁶¹Tb simulations, was subjected to grid-convergence testing and external comparison with a published DPK. Lesion dosimetry was assessed using Dmean, D90, D95, equivalent uniform dose (EUD) and tumour control probability (TCP), with uncertainty quantified using patient-cluster bootstrap confidence intervals. Kinetic sensitivity and diagnostic tracer subgroup analyses were additionally performed. Results: The study showed that ¹⁶¹Tb produced the highest median lesion-level Dmean, D90, D95 and EUD at 182.79, 136.28, 132.07 and 148.53 Gy, respectively, with a median TCP of 0.981. Corresponding values for ¹⁷⁷Lu were 141.33, 105.42, 102.10 and 114.78 Gy (TCP 0.930), while ⁹⁰Y produced lower local dose metrics but the broadest radial dose distribution, consistent with its longer-range β-particle crossfire. ¹⁶¹Tb remained the highest-ranking radionuclide across the investigated kinetic cases and within both diagnostic tracer subgroups. Hybrid ¹⁶¹Tb/⁹⁰Y kernels provided a controllable compromise between localised energy deposition and extended crossfire; a 70:30 model increased central dose localisation while retaining an R90 and R95 of 6 and 7 mm, respectively. Grid-convergence and published-DPK comparisons supported the numerical adequacy of the kernel methodology. Radionuclide emission characteristics substantially influence the transformation of heterogeneous tumour uptake into spatial absorbed-dose distributions. Within this model, ¹⁶¹Tb provided the strongest overall lesion-level dosimetric performance, whereas the extended range of ⁹⁰Y may offer complementary crossfire for selected bulky or heterogeneous lesions. Conclusions: The findings support phenotype-informed radionuclide comparison and provide a computational basis for investigating hybrid strategies. However, the absolute dose estimates and proposed radionuclide combinations remain model-based and require validation using serial therapeutic imaging, heterogeneous patient-specific dosimetry and normal-organ dose constraints before clinical translation. Full article

Microplastics are emerging as ubiquitous contaminants in agricultural soils, while nickel (Ni), although catalogued as a potentially toxic element, is an essential micronutrient and a cofactor of urease. In a pot experiment, this study examined how three microplastics (polyethylene, PE; poly(ethylene terephthalate), PET; and polystyrene, PS) affect nickel availability and the properties of two calcareous agricultural soils from central Greece. The microplastics were applied at two levels (1.5 and 3.0% by weight), and the samples were analysed at four time points (3, 6, 9, and 12 months, n = 3). Total (aqua regia) and available (DTPA) nickel, pH, electrical conductivity, organic matter, bulk density (BD), water-holding capacity (WHC), and microbial respiration (qCO₂) were measured, and the data were evaluated by one-way ANOVA with Tukey HSD tests. Total nickel remained stable (about 15.1–15.4 mg/kg; non-significant), whereas the available fraction changed systematically. In Soil 1 (12 months, 1.5%), PE raised DTPA-Ni by +24.9%, PET by +19.8% and PS by −1.7%, while at 3.0% the increases reached +39.8% and +31.7%. In parallel, PE lowered bulk density by up to −16.1%, PET raised water holding capacity by up to +28.9%, and PS raised microbial respiration by up to +38.3%, producing distinct, polymer-specific responses. The changes intensified with time and dose and were milder in the more strongly buffered Soil 2. Microplastics redistribute the biologically active, rather than the total, pool of nickel, a finding that supports reappraising Ni as a nutritional micronutrient and monitoring its available fraction. Because no plants were grown, the nutritional interpretation is advanced as a hypothesis for future testing rather than as a demonstrated agronomic benefit, and the applied microplastic doses (1.5–3.0% w/w) exceed most reported field levels and were chosen to resolve mechanisms under accelerated conditions. Full article

Botulinum toxin, once known only as the cause of botulism, has become a versatile therapeutic agent whose use in children now extends across many medical and surgical specialties. This narrative review synthesises the clinical use of botulinum toxin in patients up to eighteen years of age. We describe its pharmacology, the non-interchangeable commercial preparations, and the weight-based dosing and safety principles specific to childhood, then examine its applications by organ system. Gastrointestinal uses include functional constipation and internal anal sphincter achalasia, persistent obstruction after surgery for Hirschsprung disease, anal fissure, and achalasia. Urological uses centre on neurogenic and idiopathic detrusor overactivity and on sphincter-directed injection for dysfunctional voiding. Neurological uses are dominated by cerebral palsy spasticity and, on considerably weaker evidence, dystonia, while head and neck uses include sialorrhoea, congenital muscular torticollis, and strabismus. We also address neonatal brachial plexus palsy together with hyperhidrosis and aesthetic use in adolescents. The evidence supporting these uses is markedly uneven. Randomised controlled trials exist for only a small number of paediatric indications, chiefly spasticity in cerebral palsy, neurogenic detrusor overactivity, and chronic sialorrhoea, whereas most remaining applications rest on observational cohorts, small case series, or extrapolation from adult practice, and we grade the certainty of evidence separately for every indication. Only a few indications are formally approved, and most remain off-label, though increasingly supported. Botulinum toxin is a reversible and generally safe adjunct, but rigorous paediatric trials are still needed. Full article

Population immune pressure from vaccination and prior infection continues to drive the evolution of SARS-CoV-2. Systematic characterization of RBD mutations under complex immune backgrounds is essential for understanding viral adaptation and evolutionary trajectories. Here, we applied a deep mutational scanning (DMS) to comprehensively map the neutralization escape landscape of the Omicron variant JN.1 and its descendant lineage XEC, under immune pressure from individuals who experienced Omicron breakthrough infections following three doses of inactivated vaccines. A neutralization escape map for the single amino acid substitutions in the RBD of JN.1 or XEC was generated, and the escape efficiency of each mutation was determined. The results show that RBD escape mutations are hierarchically organized: low-intensity signals are widespread, whereas high-intensity escape is confined to a few key sites. These escape mutations are not confined solely to the receptor-binding motif (RBM) but are broadly distributed across the entire RBD. Many escape sites could accommodate multiple amino acid substitutions. Integration of DMS data with genomic surveillance of circulating variants from 2024 to 2025 revealed significant overlap between experimentally identified escape sites and mutations observed in natural isolates. This overlap increased substantially in 2025, with site concordance rising from 27.17% and 26.81% to 45.09% and 47.10% for JN.1 and XEC, respectively. The natural prevalence of these escape mutations is further shaped by factors such as receptor-binding affinity, protein stability, and epistatic interactions. Overall, our findings suggest that SARS-CoV-2 antigenic evolution follows the pattern of multiple pathways within a constrained space, providing new insights into the adaptive mechanisms of Omicron-derived variants under hybrid immune pressure. Full article

Sudden tailings dam breaches trigger large-scale heavy metal compound pollution in coupled surface water–groundwater systems, requiring systematic full-cycle ecological damage quantification tools applicable to diverse contamination types. This study constructs an integrated full-cycle ecological damage assessment framework for sudden water pollution accidents, integrating three core modules: multi-model pollutant migration prediction, multi-scale aquatic biological damage diagnosis, and three-dimensional ecological-economic loss accounting. The framework adopts a modular design that can potentially accommodate heavy metals (Cd, Cr, As, Pb) and organic pollutants such as polycyclic aromatic hydrocarbons (PAHs), with standardized molecular, individual, and population-level biological endpoints and corresponding pollutant dose–response templates reserved as reference calculation modules. However, applicability beyond this case has not been validated and requires case-specific calibration. To verify the operability and accuracy of the proposed integrated system, a typical tailings dam leakage incident dominated by hexavalent chromium (Cr(VI)) and arsenic (As) pollution was selected as the practical validation case; all field monitoring, pollutant simulation, and final economic loss quantification in this case exclusively rely on on-site measured Cr(VI) and As data, while Cd and PAH-related biological response curves and remediation cost formulas retained in the manuscript only serve as illustrative universal template components of the framework rather than case-measured results. For the Cr(VI)/As pollution case, the advection–diffusion model simulation revealed that the Cr(VI) contamination plume horizontally spread 250 m within 48 h and extended to 560 m after seven days, and anaerobic groundwater environments drove the transformation of toxic mobile trivalent arsenic (As(III)) from primary pentavalent arsenic. The calibrated SWAT model achieved Nash–Sutcliffe efficiency (NSE) coefficients of 0.75 for dissolved Cr(VI) and 0.68 for particulate As. The graph theory-based rapid prediction model cut computation duration down to minutes; when validated against independent field monitoring data, it yielded an average relative error of 14.2%, and its consistency with the SWAT model reached 10.5% relative deviation, satisfying the accuracy requirement for emergency early warning. Field biological monitoring demonstrated substantial ecological impairment: metallothionein (MT) expression in fish tissues was markedly elevated (the reported 6.2-fold induction value derives from standard Cd exposure template tests within the framework, with analogous MT upregulation also observed for field Cr(VI)/As co-stress), and benthic community Shannon diversity declined by over 50% in polluted river reaches. The standardized Ecological Damage Index (EDI) of the case was calculated as 480.2, indicating severe aquatic ecosystem damage, with total comprehensive ecological and economic losses reaching 17.25 million CNY. This study innovatively couples high-precision physical transport models with fast emergency prediction algorithms and establishes a complete multi-tier biological indicator chain linking molecular biomarkers to community integrity metrics; the three-dimensional loss accounting system integrating ecosystem service impairment, restoration expenditure, and post-pollution recovery loss realizes closed-loop full-cycle damage evaluation. The proposed framework, demonstrated for Cr(VI) and As pollution, has a modular design that may potentially be extended to other pollutants such as Cd and PAHs by adjusting model parameters, providing a quantitative reference for emergency disposal, pollution remediation, and ecological compensation of water contamination accidents, although further validation across different pollutants and hydrological settings is required. Full article

Heavy metals in water are a global concern due to their persistence, toxicity, and bioaccumulation potential, highlighting the need to assess their effects on aquatic organisms. The ecotoxicity of eight potentially toxic metal nitrates (Cd (0.002 mg/L), Co (0.02 mg/L), Cr (0.02 mg/L), Cu (0.001 mg/L), Mn (0.04 mg/L), Ni (0.005 mg/L), Pb (0.002 mg/L), Zn (0.1 mg/L) up to 1000 mg/L for all metals) was assessed on the aquatic macrophyte Lemna minor, in a multilevel study. Growth responses, measured as frond number and average specific growth rate, were used to generate dose–response curves, but also to calculate EC₅₀ (Median Effective Concentration) and E_rC₅₀ (50% Effect Concentration based on growth rate) values. Based on EC₅₀, toxicity followed the order Cd > Co > Ni > Zn > Cu > Cr > Pb > Mn, while E_rC₅₀-based ranking was Cd > Ni > Co > Zn > Cu > Cr > Pb > Mn. Metals were classified as highly toxic (Cd, Ni; Co based on EC₅₀), moderately toxic (Co based on E_rC₅₀, Zn based on EC₅₀), slightly toxic (Cu, Cr; Zn based on E_rC₅₀, Pb based on EC₅₀), or practically non-toxic (Pb based on E_rC₅₀, Mn). Biochemical responses regarding photosynthetic pigments, primary metabolites, and oxidative-stress-related enzymes were investigated, revealing physiological effects for most metals. Species sensitivity distribution-based comparison indicated that L. minor sensitivity varies across metals, ranging from typical to relatively low within the broader species sensitivity distribution. Although the toxicity of potentially toxic metals to L. minor has been extensively investigated, data on ErC50 values for several of the metals investigated in the present study remain limited, highlighting the need for further ecotoxicological research. Full article

Background/Objectives: Microplastics (MPs) and nanoplastics (NPs) have emerged as ubiquitous environmental contaminants resulting from the extensive production, use, and degradation of plastic materials. Human exposure occurs primarily through contaminated food and drinking water, with inhalation representing an additional important route. Growing concern has focused on the ability of these particles, particularly NPs, to cross biological barriers, enter the systemic circulation, and reach human tissues. The aim of this review was to summarize current evidence on dietary exposure to MPs and NPs, their gastrointestinal bioavailability and toxicokinetics, and their potential systemic health effects, with particular emphasis on organ-specific responses, underlying biological mechanisms, and the strength and limitations of the available evidence. Methods: A comprehensive narrative review of the scientific literature published between 2000 and 2026 was conducted using PubMed/MEDLINE, Scopus, Web of Science, and Google Scholar. Original research articles and review papers addressing dietary exposure, occurrence in food and drinking water, migration from food-contact materials, gastrointestinal absorption, translocation, biodistribution, bioaccumulation, elimination, molecular mechanisms, and potential organ-specific or systemic health effects were included. Publications without full-text availability, conference proceedings, editorials, commentaries, duplicate publications, and studies without relevance to human exposure or health were excluded. Results: Food, drinking water, beverages, and food-contact materials represent important sources of human exposure to MPs and NPs. Following ingestion, most larger particles are eliminated through the gastrointestinal tract, whereas smaller MPs and particularly NPs may cross biological barriers and potentially reach the systemic circulation and distant tissues. Experimental studies consistently identify interconnected biological responses involving oxidative stress, inflammation, mitochondrial dysfunction, barrier impairment, immune dysregulation, genotoxicity, apoptosis, and endocrine disruption. These mechanisms have been associated with alterations in the gastrointestinal, respiratory, cardiovascular, nervous, urinary, reproductive, endocrine, and skeletal systems and with biological processes relevant to carcinogenesis. However, most mechanistic evidence derives from in vitro and animal models, whereas human evidence remains limited and predominantly observational. Consequently, the extent to which these experimental findings translate into clinically significant effects in humans remains uncertain. Conclusions: Current evidence supports the biological plausibility of systemic effects associated with MNP exposure but is insufficient to establish causal relationships between chronic dietary exposure and specific human diseases. The detection of MNPs in human tissues and reported associations with pathological conditions should therefore be interpreted cautiously. Standardized analytical methods, improved characterization of realistic human exposure, and well-designed longitudinal epidemiological studies integrating quantitative exposure assessment with validated clinical outcomes are required to clarify dose–response relationships, long-term health effects, and the clinical significance of MNP exposure. Full article

Direct-acting antiviral (DAA) therapy has dramatically transformed the management of hepatitis C virus (HCV) infection in chronic kidney disease (CKD). Severe renal impairment and dialysis dependence are no longer considered major barriers to virologic cure. The main difficulties have shifted toward efficient diagnosis, choosing the regimen in the presence of cirrhosis and drug interactions, coordinating treatment with kidney transplantation, managing HCV-associated immune-complex kidney disease, and providing treatment in dialysis and resource-limited settings. This narrative review aims to discuss these issues in a decision-oriented manner, using verified guidelines, systematic reviews, important clinical trials, transplant cohorts, and studies of cryoglobulinemic disease, while explicitly comparing the strength and consistency of the underlying evidence and highlighting areas of genuine clinical uncertainty. Present evidence supports using the recommended DAA regimens without renal dose adjustment, while ribavirin needs dose modification when kidney function is reduced and can result in hemolytic anemia. Kidneys from HCV-viremic donors can be transplanted to recipients without HCV infection when proper informed consent, rapid access to DAA treatment, and organized post-transplant monitoring are available, although the minimum effective duration of peri-transplant antiviral prophylaxis remains unresolved. Antiviral therapy is the first-line treatment for HCV-associated glomerular disease, while rituximab-based immunosuppression is largely reserved for severe, rapidly progressive, or persistent cryoglobulinemic vasculitis. Future progress will depend less on proving antiviral efficacy and more on closing the gaps between screening, confirmatory testing, starting treatment, transplantation pathways, and long-term follow-up, gaps that stem from inequities in diagnostic infrastructure, drug reimbursement, and healthcare-system organization as much as from any remaining biomedical uncertainty. Full article

Water deficit is one of the main limiting factors for crop establishment and productivity, particularly during seed germination and early seedling development. This study aimed to evaluate the biostimulant potential of Ascophyllum nodosum extract applied to maize (Zea mays L.) seeds subjected to osmotic stress induced by polyethylene glycol (PEG-6000). Three independent bioassays were conducted under controlled conditions. In the first assay, osmotic potentials ranging from 0 to −0.8 MPa were evaluated to characterize stress levels and identify conditions representative of moderate and severe water deficit. In the second assay, increasing doses (0 to 2.0 mL kg⁻¹ of seeds) of a commercial A. nodosum-based extract and its isolated mineral fraction were evaluated to determine the optimal application range. In the third assay, the selected treatment was evaluated under control conditions and under moderate (−0.6 MPa) and severe (−0.8 MPa) osmotic stress. Germination percentage, normal and abnormal seedlings, radicle and epicotyl length, and seedling vigor index were assessed. Osmotic stress progressively reduced germination and seedling growth, with the strongest effects observed at −0.6 and −0.8 MPa. Seed treatment with A. nodosum extract did not impair final germination but improved seedling development in a dose-dependent manner. The highest efficiency was observed at intermediate doses, with maximum normal seedling percentage predicted at 0.45 mL kg⁻¹ and maximum radicle growth at approximately 0.66 mL kg⁻¹. Under water deficit conditions, the complete extract promoted greater improvements in seedling performance than the mineral fraction, indicating that the beneficial effects were mainly associated with bioactive organic compounds rather than mineral nutrition alone. These findings provide new evidence that dose optimization is essential for maximizing the efficiency of seaweed-based biostimulants and demonstrate the potential of A. nodosum seed treatment as a sustainable strategy to improve maize establishment under water-limited conditions. Full article

Radiation-induced liver disease (RILD) remains a clinically significant complication of radiotherapy for malignant neoplasms of the liver and upper abdomen, restricting achievable therapeutic doses and adversely affecting patient outcomes. Conventional photon-based radiotherapy (X-rays/γ-rays) exposes substantial volumes of healthy liver parenchyma and adjacent organs to ionizing radiation, increasing the risk of hepatocellular damage, inflammation, and progressive fibrosis. Electron irradiation (β-particles) represents a promising alternative owing to its limited tissue penetration (~2–3 cm) and steep dose fall-off, which significantly reduces exit dose and may reduce exit dose and limit exposure of paratumoral healthy tissues compared with photon therapy. However, the molecular mechanisms underlying electron-induced hepatic damage and strategies for its prevention remain insufficiently characterized. This review systematically analyzes the pathogenesis of radiation-induced liver damage, encompassing direct DNA damage, oxidative stress, mitochondrial dysfunction, NF-κB-mediated inflammation, cellular senescence, and TGF-β/Smad-driven fibrosis. The comparative dosimetric and radiobiological advantages of electron irradiation over photon therapy are discussed, with particular attention to intraoperative radiotherapy, FLASH, and very high-energy electron techniques. Current radioprotective strategies are critically evaluated, with emphasis on the limitations of amifostine and the multifunctional radioprotective potential of ascorbic acid, including free radical scavenging, attenuation of lipid peroxidation, stimulation of endogenous antioxidant defense, and direct inhibition of radiation-induced DNA strand breaks. Existing gaps in the evidence base are identified, and recommendations for future experimental and clinical research are proposed. Full article

Aerobic composting converts agricultural waste into stable, humus-rich products, and the application of exogenous additives is an efficient strategy to enhance humification. This systematic review synthesized studies published between 2023 and 2026 on additives that promote humification during agricultural waste composting. Based on the distribution of the retrieved literature, additives are categorized into inorganic additives, organic additives, biological strategies, and composite systems, and the effects and mechanisms of each category are systematically discussed. Iron-based additives achieve the highest humic acid (HA) increases of 82–267% through Fenton-like redox catalysis. Clay minerals and biochar produce moderate HA enhancements of 25–163% via physical structuring and surface adsorption with broader applicability. Small-molecule precursors and exogenous humic substances achieve HA gains exceeding 100% at sub-percent doses. Biological strategies provide self-sustaining catalytic activity but are sensitive to environmental conditions. Composite additives, the largest category, generally outperform single additives through functional complementarity, though antagonistic effects have also been documented. Cross-study patterns suggest that different feedstocks respond preferentially to distinct additive types, though systematic experimental validation is lacking. Critical gaps between laboratory findings and practical application are identified, including the predominance of small-scale studies, the absence of techno-economic analysis, and the unassessed environmental fate of metal-based additives. Future research priorities include pilot-scale validation under industrial conditions, the establishment of standardized humification metrics, and long-term field monitoring. Full article

Background: The clinical translation of cell-based therapies for knee articular cartilage repair is fundamentally restricted by the severe biological unpredictability of autologous cell sources, inherent manufacturing bottlenecks, and the rapid phenotypic dedifferentiation of cells expanded in conventional 2D monolayers. To overcome these translational hurdles, this study engineered a scaffold-free, 3D formulation of highly characterized allogeneic FE002-Cart chondroprogenitor spheroids. Methods: We systematically investigated the specific microenvironmental cues and Good Manufacturing Practice (GMP) formulation parameters required to direct functional chondrogenesis. The structural and biochemical performance of this allogeneic formulation was benchmarked against multiple primary adult autologous chondrocyte types. Finally, we evaluated the phenotypic resilience of the microtissues in simulated osteoarthritic (OA) environments and investigated both short-term liquid storage and advanced terminal preservation strategies to establish off-the-shelf logistical viability. Results: Precise microenvironmental regulation proved to be a critical biological prerequisite. The synergistic combination of physiological hypoxia (2% O₂) and stringent glucocorticoid limitation (10 nM dexamethasone) induced robust glycosaminoglycan (GAG) deposition and a > 200-fold upregulation of ACAN and COL2, while suppressing the terminal hypertrophic drift observed in adult chondrocytes. Benchmarking revealed that the allogeneic FE002-Cart formulation substantially mitigates the profound morphological and biochemical unpredictability inherent to adult autologous cell sources. Furthermore, the scaffold-free spheroid geometry yielded a 10-fold increase in GAG production per cell compared to traditional matrix-seeded (MACI) platforms. Transitioning to a GMP-compatible manufacturing process revealed extreme cellular sensitivities; excipients within standard pharmaceutical-grade dexamethasone severely aborted chondrogenic differentiation, emphasizing the necessity of rigorous raw-material qualification. Functionally, the 3D architecture acted as a protective physical shield, sustaining high cellular viability when subjected to severe inflammatory stress and 100% OA patient synovial fluid. Logistically, the viable spheroids maintained matrix integrity and inter-spheroid fusion potential for up to 7 days at ambient temperature in transport medium. Finally, advanced spheroid preservation via lyophilization and high-dose gamma irradiation eliminated biological viability but successfully transitioned the microtissues into highly organized, terminally irradiated matrices capable of heterologous in vitro structural merging. Conclusions: These findings define the critical biological thresholds for manufacturing, demonstrate the enhanced in vitro biosynthetic efficiency of 3D allogeneic microtissues compared to specific autologous and matrix-dependent baselines, and establish a highly practical, off-the-shelf logistical framework for the regenerative treatment of large knee chondral defects. Full article

Intracranial abscesses are extremely rare in alpacas, and previously reported cases have lacked systematic etiological characterization. The virulence characteristics of Mammaliicoccus sciuri (formerly Staphylococcus sciuri) in alpaca intracranial infections remain unclear. We investigated an alpaca with chronic otitis media and an intracranial abscess. Bacteria were recovered from intracranial purulent material and several other sites. A representative isolate was designated CS234. It was identified as M. sciuri by 16S rDNA sequencing and whole-genome sequencing (WGS). Average nucleotide identity (ANI) analysis showed 95.58% identity with M. sciuri NCTC12103. WGS identified mecA, mecA1, fexA, optrA, and sal(A). The isolate showed phenotypic resistance to aminoglycosides, tetracyclines, and macrolides. PCR screening detected several virulence-associated gene targets, including exhB and tsst-1. To our knowledge, this is the first report of exhB in M. sciuri. In an intraperitoneal mouse challenge, the median lethal dose (LD₅₀) was 4.4 × 10⁷ colony-forming units (CFU)/g body weight. Multiple-organ lesions were also observed. These findings indicate systemic pathogenic potential in mice. Recovery of M. sciuri from several affected sites indicates a strong etiological association with the suppurative disease in this alpaca. These findings expand current knowledge of M. sciuri infection and antimicrobial resistance in alpacas. They may also aid the diagnosis and treatment of similar cases. Full article