Commonly used accuracy metrics for non-rigid registration such as target registration error (TRE) do not assess the mechanical plausibility of predicted deformations. We introduce a metric to quantify local coherence of deformation and complement existing measures of physical validity. We propose the strain alignment metric, which evaluates local directional consistency of deformation by analyzing the principal directions and magnitudes of the Green strain tensor. The metric combines three components: (i) a sign term penalizing local flips between compression and extension, (ii) a magnitude term capturing relative variation in principal strain magnitude within a neighborhood and (iii) an angular term measuring non-directional alignment of principal strain directions. The metric is computed locally and summarized over the mesh. We compare it to the Jacobian determinant and strain norm on synthetic liver deformations registered using a deep learning-based method and a fast biomechanical FEM approach. While both methods achieve comparable TRE, the proposed metric captures substantial differences in deformation coherence on the sample level that the Jacobian determinant and strain norm only show upon single mesh inspection. It is able to clearly differentiate the largely coherent deformation despite volume changes of the biomechanical method from the local inconsistencies of the learning-based method, including alternating compression and extension patterns. Strain alignment provides complementary, interpretable information about the mechanical plausibility of predicted displacement fields. It enables quantitative assessment of local deformation coherence without requiring material assumptions and further highlights limitations of accuracy-only evaluation in non-rigid registration.
Lignin-rich industrial streams represent an abundant but underutilized source of renewable aromatic carbon. Efficient biological conversion requires microbial hosts capable of metabolizing chemically diverse lignin-derived aromatic compounds; however, such capabilities are typically found in environmental bacteria, which are constrained by physiological and metabolic limitations. Sphingobium lignivorans SYK-6 harbors extensive aromatic catabolic pathways, but its inability to utilize glucose and its methionine auxotrophy have limited its use as a production host. Here, we identified the metabolic basis of these constraints and systematically rewired the underlying pathways to overcome them. Introduction of a heterologous glucose transporter, reconstruction of methionine biosynthesis, chromosomal integration of pathway genes, and adaptive laboratory evolution collectively enabled robust growth on glucose while eliminating methionine auxotrophy. The engineered strain converted lignin-derived aromatics in oxygen-soda-anthraquinone pulping black liquor derived from Japanese cedar, achieving high-yield production of the polymer precursor 2-pyrone-4,6-dicarboxylic acid (PDC) (2.71 g/L and >130 mol% conversion relative to major quantified aromatics). We further show that gluconolactonase can substitute for 6-phosphogluconolactonase in the Entner-Doudoroff pathway, demonstrating that central carbon metabolism can accommodate functionally analogous enzymes. Together, these results provide a metabolically rewired SYK-6 strain as a platform for the valorization of industrial lignin streams and suggest that overcoming physiological and metabolic constraints can enable non-model aromatic-degrading bacteria to function as industrial production hosts.
Flexible strain sensors are essential components for wearable electronics, implantable biointerfaces, and soft human-machine systems. As application scenarios expand from epidermal monitoring toward long-term in vivo operation, increasingly stringent requirements are imposed on multifunctionality, mechanical compliance, signal stability, and biointegration. Carbon-based functional materials, owing to their tunable electrical properties, structural versatility, and favorable electromechanical compatibility, have emerged as a central materials platform for next-generation flexible strain sensors. This review presents a comprehensive, mechanism-oriented overview of carbon-enabled flexible strain sensing, encompassing piezoresistive, capacitive, and piezoelectric transduction modes. This review systematically examine how carbon material dimensionality including low-dimensional nanofillers, two-dimensional sheets, and three-dimensional porous, governing sensitivity, durability, and long-term device reliability. Particular emphasis is placed on contrasting the fundamentally different design requirements of wearable and implantable systems, including sensitivity-stability trade-offs, tissue-level mechanical matching, and operational robustness in complex biological environments. Distinct from prior material- or device-centric reviews, this review establishes a unified framework linking carbon architectures, sensing mechanisms, and application contexts, thereby clarifying critical bottlenecks and design principles for advancing multifunctional, biointegrated strain sensors toward practical and translational use.
Lead (Pb) contamination in mining tailings from Zaruma, Ecuador, represents an environmental concern due to its toxicity, persistence, and potential mobility under changing physicochemical conditions. This study evaluated Pb mobilization from mining tailings mediated by a Bacillus safensis group strain under controlled laboratory conditions. A central composite design (CCD) was applied to assess the influence of initial pH, temperature, incubation time, and solids concentration, while kinetic modeling, pH evolution analysis, apparent Gibbs-energy-based assessment, and generalized additive modeling (GAM) were used to interpret the process. Maximum Pb mobilization reached 1278.76 mg kg⁻1, equivalent to 96.51% release, at pH 5.75, 31 °C, 14.5 days, and 0.015% solids. Pb release kinetics were best described by the shrinking core model (R2 = 0.99), suggesting diffusion through an apparent product layer as the main rate-limiting step during later stages. pH evolution followed an exponential decay model with plateau (R2 = 0.99), indicating progressive microbial acidification and subsequent stabilization. AICc-based GAM comparison selected the two-predictor pH-temperature interaction model as the final parsimonious structure, with 62% explained deviance and reduced overfitting risk under the limited sample size. The apparent Gibbs-energy-based assessment indicated that Pb mobilization was governed by microbial activity, pH evolution, and mass-transfer conditions rather than by global reaction spontaneity alone. These results suggest that the Bacillus safensis group strain can substantially influence Pb mobility in Zaruma tailings under favorable low-solid laboratory conditions, although validation at higher solids loadings and Pb recovery from the liquid phase are required for practical application.
The acquisition efficiency of functional Haloarchaea strains using traditional dilution-plating methods remains low. To address this, we established a targeted isolation strategy for screening diverse and active functional Haloarchaea strains through single-cell manipulation at the microscopic level, based on selective substrates, cell division inhibition, and fluorescent staining.
Two novel types of dimeric alkaloids were isolated from the marine sediment-derived mutant strain Saccharopolyspora erythraea SCSIO 07745/Δspo11. Among them, sacchaindol A (1) represents a previously unreported class of dimeric aminoquinolinone alkaloids, whereas sacchaindols B (2) and C [(±)-3] comprise a rare fused 6/6/5/5 tetracyclic indole alkaloid framework. Their structures were elucidated using a combination of spectroscopic analyses, single-crystal X-ray diffraction, and computational methods. Hypothetical biosynthetic pathways for 1-3 were proposed. Notably, sacchaindols C [(±)-3] exhibited anti-inflammatory activity.
Photovoltaic wastewater often contains hexavalent chromium (Cr(VI)), fluoride (F-), and poorly biodegradable polyethylene glycol (PEG), and this complex pollution poses a severe challenge to biological denitrification systems. This study focused on strain LX-6 (Acinetobacter sp.), which possesses both heterotrophic nitrification-aerobic denitrification (HNAD) and microbial-induced calcium precipitation (MICP) capabilities, to systematically investigate the inhibitory effects of Cr(VI) on denitrification and mineralization and to elucidate the mechanism by which PEG alleviates Cr(VI) stress through heterogeneous nucleation regulation. Cr(VI) significantly inhibited biomass, electron transport system activity (ETSA), HNAD enzyme activity, and mineralization efficiency in a concentration-dependent manner. PEG coordinated with calcium via its ether bonds and hydroxyl groups, providing additional heterogeneous nucleation sites and acting synergistically with extracellular polymeric substances to construct a composite nucleation interface, thereby promoting the rapid formation of a dense mineralized shell on the bacterial surface during the early stage of the reaction. This shell acted as a diffusion barrier and effectively mitigated the direct cytotoxicity of Cr(VI). Compared with the Cr(VI)-only treatment, PEG increased adenosine-5'-triphosphate by 16.21% and ETSA by 92.86%, decreased malondialdehyde by 47.52%, and significantly enhanced antioxidant enzyme activity. The electrochemical impedance characteristics showed significant improvement, with the limiting current recovering from 0.287 mA to 0.562 mA. Cr(VI) was ultimately bioreduced and immobilized as Cr(III) precipitates, whereas PEG is primarily removed by adsorption and coprecipitation. This study provides new insights into the mechanisms and technical approaches for the synergistic advanced treatment of photovoltaic composite wastewater containing Cr(VI), F-, and recalcitrant PEG.
Elsinoë species are slow-growing, hemibiotrophic to necrotrophic fungi that cause scab diseases on economically important fruit crops. Genome resources for many host-specific species remain limited. We report high-quality draft genome assemblies for the ex-type strains of Elsinoë mangiferae (CBS 226.50) and E. perseae (CBS 406.34), causal agents of mango and avocado scab, respectively. Among five approaches tested, a Nanopore-only NextDenovo assembly produced the most contiguous genomes, yielding 24.5 Mb (E. mangiferae) and 25.1 Mb (E. perseae) assemblies with 13 and 18 contigs, respectively, BUSCO completeness scores of ∼94%, and multiple putative telomere-to-telomere chromosomes. Gene prediction identified 9,134 and 9,243 genes, respectively. Functional annotation revealed enrichment of metabolic and regulatory pathways, including those involved in posttranslational modification, protein transport, and secondary metabolism. Carbohydrate-active enzyme repertoires were small but conserved, consistent with stealth pathogenicity strategies and low plant cell wall degradation. Both genomes encoded large secretomes (>850 proteins), diverse protease repertoires (>300 proteins), Ecp2-like effector proteins, and multiple biosynthetic gene clusters, including clusters with similarity to those associated with elsinochrome and ACT-toxin II biosynthesis, some of which may contribute to host-pathogen interactions and disease development. A large fraction of genes lacked functional characterization, suggesting incomplete databases and/or the presence of lineage-specific genes potentially involved in virulence or host adaptation. These genome resources fill critical gaps for underrepresented Elsinoë species and provide taxonomically anchored references essential for diagnostics, comparative genomics, and research into the molecular basis of host specificity and pathogenicity in scab-causing fungi.
1,3-dioxolane is a promising solvent for low-temperature batteries owing to its low freezing point and low viscosity. However, its tendency toward ring-opening polymerization leads to reduced ionic conductivity and deteriorated electrochemical stability. Here, we establish an electronic-geometric coupling design principle to regulate solvent stability in weak-weak electrolyte systems for sodium metal batteries. A dual-descriptor framework combining ring strain energy (RSE) and a sterically corrected electrostatic descriptor, defined by the lowest negative electrostatic potential normalized by molecular volume (ESPmin/Volume), is introduced to guide cyclic ether solvent design. Following this principle, 2,4-dimethyl-1,3-dioxolane is identified with reduced RSE and moderate ESPmin/Volume, enabling enhanced resistance to polymerization and improved Na-compatibility/ion transport. Molecular dynamics simulations and density functional theory calculations reveal that, the electrolyte forms an aggregate-dominated solvation structure with a high lowest unoccupied molecular orbital level, promoting the formation of a thin, uniform, and inorganic-rich solid electrolyte interphase. Consequently, the electrolyte delivers accelerated interfacial kinetics and stable operation across a wide temperature range. Na||Na symmetric cells cycle stably for 1800 h at room temperature, while Na||Na3V2(PO4)3 full cells with high cathode loading (20 mg cm-2) operate for over 200 cycles at 25 °C and more than 900 cycles at -40° C.
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In this study, three novel actinomycete strains, designated R6R-6ᵀ, R7R-8ᵀ and R16R-3ᵀ, were isolated from the roots of Glycosmis pentaphylla (Retz.) DC., Aegle marmelos (L.) Corrêa ex Roxb. and Citrus × aurantiifolia, respectively. The taxonomic status of these strains was evaluated using a polyphasic taxonomic approach. All three strains produced short spore chains (three to five spores) with cylindrical spores on aerial mycelia and exhibited chemotaxonomic characteristics consistent with members of the genus Nocardia. These features included the presence of meso-diaminopimelic acid and mycolic acids in the cell wall peptidoglycan, a polar lipid profile of type PII characterized by phosphatidylethanolamine, and cellular fatty acids (>1%) comprising C14 : 0, C16 : 0, C17 : 0, C18 : 0, C18 : 0 10-methyl, C16 : 1 ω11c, C18 : 1 ω9c and C16 : 1 ω6c and/or C16 : 1 ω7c. Based on 16S rRNA gene sequence analysis, strains R6R-6ᵀ and R7R-8ᵀ showed the highest sequence similarities of 98.9 and 99.1%, respectively, to Nocardia xestospongiae ST01-07ᵀ, while strain R16R-3ᵀ exhibited the highest similarity (98.5%) to Nocardia anaemiae NBRC 100462ᵀ. Genome-based comparisons using digital DNA-DNA hybridization and average nucleotide identity revealed that all three strains shared values below the established species delineation thresholds when compared with their closest phylogenetic relatives. Based on the combined phenotypic, phylogenetic and genomic evidence, strains R6R-6ᵀ, R7R-8ᵀ and R16R-3ᵀ represent three novel species of the genus Nocardia, for which the names Nocardia glycosmidis sp. nov. (type strain R6R-6ᵀ=TBRC 14548T, =NBRC 115197T), Nocardia aegles sp. nov. (type strain R7R-8ᵀ=TBRC 14549T, =NBRC 115198T) and Nocardia citri sp. nov. (type strain R16R-3ᵀ=TBRC 14550T, =NBRC 115199T) are proposed.
A novel α-haemolytic bacterial strain, IMAU11619T, belonging to the genus Streptococcus, was isolated from human intestinal tract samples collected in Hohhot City, Inner Mongolia Autonomous Region of China. Strain IMAU11619T is characterized as Gram-positive, non-motile, non-spore-forming cocci. Optimal growth conditions were anaerobically determined at 37 °C and pH 6 in De Man-Rogosa-Sharpe medium. We sequenced the draft genome of strain IMAU11619T and characterized its taxonomic status using phylogenetic and phenotypic analyses. Comparative analysis with the TYGS database indicated that strain IMAU11619T represents a novel species within the genus Streptococcus that is phylogenetically distinct from all previously characterized species. Phylogenetic reconstruction based on 176 single-copy genes positioned this strain within the parasanguinis subclade of the Mitis-Suis clade and closely related to Streptococcus rubneri DSM 26920T, Streptococcus australis ATCC 700641T and Streptococcus ilei I-G2T. The highest observed average nucleotide identity and digital DNA-DNA hybridization values with its closest relative, S. australis ATCC 700641T, were 93.1-93.8 and 52.8%, respectively. The genomic DNA G+C content of strain IMAU11619T was 42.2 mol%. The major fatty acid components identified included C16:0, C14:1, C18:0 and summed feature 8 (C18:1  ω7c and/or C18:1  ω6c). The respiratory quinone type of strain IMAU11619T was identified as menaquinone-9. Based on its distinctive phylogenetic position and phenotypic features, strain IMAU11619T is proposed as a novel species, Streptococcus intestinihominis sp. nov., with type strain IMAU11619T (=CGMCC 1.64731T=GDMCC 1.4355T=JCM 37062T).
Iron deficiency is the leading cause of anemia in childhood and may induce cardiovascular adaptations, including increased heart rate, elevated cardiac output, plasma volume expansion, and myocardial remodeling. Beyond hematologic consequences, reduced iron availability may also affect myocardial structure and function. This study aimed to assess biventricular function in children with iron deficiency anemia using two-dimensional speckle-tracking echocardiography. This prospective case-control study included pre- and post-treatment evaluations. Forty children with iron deficiency anemia and 29 healthy controls were enrolled. Of the 40 patients, 24 were re-evaluated after treatment. Tissue Doppler imaging was used to measure myocardial velocities, isovolumic contraction time, isovolumic relaxation time, and ejection time at the interventricular septum and the basal segments of both ventricles. Speckle-tracking echocardiography was used to assess left ventricular longitudinal strain (LS) and strain rate, left ventricular circumferential strain and strain rate, as well as right ventricular global longitudinal strain and strain rate. Left and right ventricular LS values improved following treatment, indicating recovery of myocardial function. Tissue Doppler parameters also demonstrated improvement in both systolic and diastolic function. Although significant improvement was observed after treatment, some echocardiographic parameters did not fully normalize, suggesting that subtle myocardial alterations may persist. These findings should be interpreted with caution due to the relatively small sample size and the predominance of mild-to-moderate anemia in the study population. Children with iron deficiency anemia may benefit from longer-term follow-up and post-treatment evaluation.
Microplastic (MPs) pollution poses a global threat to ecosystems and human health. Traditional physicochemical methods face limitations like high cost, inefficiency, and secondary pollution, underscoring the need for green remediation. While microbial degradation holds promise, research remains largely limited to lab studies of single environments, lacking systematic multi-environment strain collections and practical application assessments. This review summarizes common screening/validation methods, degradation mechanisms, and influencing factors of microplastic-degrading microorganisms (MPDMs). By reviewing 78 studies, 197 degrading microorganisms/taxa from four typical environments (soil, marine, freshwater, animal gut) were sorted, with 152 analyzed for degradation efficiency. The degradation efficiencies across different environments were compared, with the maximum efficiency order being soil > freshwater > marine > animal gut, and the overall average efficiency order being soil ≈ freshwater > marine ≈ animal gut. Recommendations for efficient culture conditions were proposed for different environments, and high-potential strains were recommended, including multi-environment efficient ones (e.g., Bacillus and multiple bacterial consortia) and environment-specific superior strains (e.g., Rhodococcus, Oceanimonas, and Enterobacter). Finally, methods such as dominant strain screening, acclimatization, genetic engineering modification, and synthetic taxa construction via machine learning are proposed to enhance practical applicability. This study not only establishes a scenario-specific microbial remediation resource pool and quantifies their degradation efficiencies, providing critical data support and strategic guidance for the development of in-situ microbial remediation technologies at targeted contaminated sites, but also lays a solid foundation for translating laboratory findings into practical field applications in future research.
Kauniolide, a parthenolide derivative, serves as a key biosynthetic intermediate for various guaianolide-type sesquiterpenoids, such as agrabin, lactucin, and lactupicrin. However, its chemical synthesis is often hampered by demanding reaction conditions and high reagent consumption, which severely limit further development and application. To address this, this study employed systematic metabolic engineering strategies to construct an efficient yeast cell factory for kauniolide production. First, the biosynthetic pathway genes for costunolide-the direct precursor of kauniolide(HaGAS, TpGAO, TpCOS, AaCPR)-were heterologously expressed in a lab-engineered yeast chassis strain with high farnesyl pyrophosphate(FPP) production. This initial strain produced costunolide at a titer of 6.1 mg·L~(-1). Subsequently, the co-expression of AaADH1, AaALDH1, and AaCYB5 was implemented to enhance the germacrene A acid synthesis and promote electron transfer, thereby boosting the catalytic efficiency of the key cytochrome P450 enzymes. This modification increased the costunolide titer to 71.5 mg·L~(-1). To optimize the conversion of costunolide to kauniolide, a promoter compatibility screen for the TpKLS gene was conducted. Among the tested promoters(P_(GAL1), P_(TDH3), P_(TEF1), P_(TPI1), P_(sptGAL2), and P_(skGAL2)), the strain harboring the P_(skGAL2)-driven TpKLS construct showed the highest performance, achieving a kauniolide titer of 28.2 mg·L~(-1). Furthermore, to further enhance the pathway flux, the catalytic efficiency of the TpKLS enzyme was improved through semi-rational design coupled with computer-aided design. The best-performing mutant, TpKLS~(V204R), exhibited a 2.7-fold higher catalytic activity compared to the wild-type enzyme. The final engineered strain, when cultivated in shake-flask fermentation, produced costunolide and kauniolide at titers of 35.0 and 71.1 mg·L~(-1), respectively. The kauniolide titer represents the highest level reported to date in a yeast system. In conclusion, this study successfully constructed an efficient yeast cell factory for kauniolide by reconstructing and optimizing its heterologous biosynthetic pathway. It provides a solid technical foundation and a valuable reference for the scalable biosynthesis of kauniolide and the exploration of its downstream derivatives.
In this research we localize the cytokinesis-inhibiting sepD5 mutation in Aspergillus nidulans to gene AN3659, previously named paxB, which is predicted to encode a LIM domain protein orthologous to the Pxl1 scaffold proteins of S. cerevisiae and S. pombe. The genetic lesion in sepD5 is predicted to result in a Q-to-R amino acid substitution at position 31 of the 776-residue PaxB protein within a region of intrinsic disorder. When grown at restrictive temperature, sepD5 strains are aseptate with impaired conidiogenesis. AN3659 null mutants replicate the sepD5 phenotype. Neither the sepD5 mutation nor deletion of paxB prevents assembly of the cortical contractile actomyosin rings (CARs) at putative septation sites, though the rings fail to constrict. Fluorescently-tagged wild type PaxB localizes to both the CAR and the Spitzenkörper (Spk) at the growing hyphal apex. Through structural truncation studies, we identify specific regions of PaxB whose presence is necessary for localization to the CAR and the Spk, as well as for proper formation of conidiophores. Deletion or downregulation of genes that prevent formation of actin rings at septation sites also blocks localization of PaxB to those same sites. Deletion or downregulation of genes that permit cortical actin ring formation while blocking constriction of those same rings does not prevent PaxB localization to CARs. When grown at restrictive temperature, sepD5 strains and paxB null strains permit normal targeting of several septation-associated proteins to cortical actin rings, while recruitment of the formin SepA, the serine-threonine protein kinase PkcA, and the chitin synthase ChsA is impaired.
This study aimed to explore emergency medical dispatchers' (EMDs') experiences of prioritising patients and stewarding ambulance resources when system capacity was constrained. Qualitative interview study using inductive qualitative content analysis. Emergency medical communication centres (EMCCs) in Sweden, operated by the national emergency call provider and responsible for receiving 112 calls and dispatching ambulances. 13 purposively sampled EMDs with at least 1 year of professional experience. Interviews were analysed inductively using qualitative content analysis (Elo and Kyngäs) through open coding, grouping into subcategories and abstraction into generic categories and one main category. Dispatchers described prioritisation under scarcity as system work that simultaneously addressed individual patient acuity and population-level readiness. One main category captured this work: stewarding scarce response capacity. Three inter-related generic categories characterised stewardship: (1) prioritising by clinical urgency within geographic and operational constraints; (2) producing availability through anticipation, reassessment and queue governance in a 'virtual waiting room'; and (3) coordinating response through information infrastructures and interprofessional collaboration. Across categories, dispatchers described redistributing risk across patients and time, managing moral strain when delays could harm patients and using experience, reassessment and teamwork to avoid both under-response to urgent need and over-allocation that would leave areas without coverage. Dispatch under scarcity is best understood as active stewardship of a safety-critical dispatch queue. Strengthening patient safety therefore requires organisational support for reassessment and escalation during prolonged waits, explicit governance of queue dynamics and geographic coverage trade-offs, safeguards for contextual judgement when automation is used and support for dispatchers exposed to morally difficult scarcity decisions.
Legumes establish a mutualistic interaction with nitrogen-fixing rhizobia. Lotus japonicus is a model for studying this symbiosis; however, only a limited number of rhizobial species nodulating this host have been taxonomically described. Here, we characterise four Mesorhizobium strains (DC-1.1T, Qj1B1, DC-1.5T, and Qj2B2) isolated from root nodules of Lotus japonicus and Lotus burttii. Multi-locus phylogeny and phylogenomic analyses resolved these isolates into two well-supported monophyletic clades. Genome-based comparisons supported their classification as distinct taxa, with strains DC-1.1T and Qj1B1 showing 95.2% average nucleotide identity (ANI) and 62.9-63.5% digital DNA-DNA hybridisation (dDDH) values relative to Mesorhizobium newzealandense ICMP 19545T, whereas DC-1.5T and Qj2B2 exhibited 92.5-92.8% ANI and 49.9-50.5% dDDH compared with Mesorhizobium waimense ICMP 19557T. Together with chemotaxonomic and physiological traits, these data support the proposal of two novel species, Mesorhizobium bavaricum sp. nov. (DC-1.1T and Qj1B1) and Mesorhizobium monacense sp. nov. (DC-1.5T and Qj2B2). Metagenomic analyses predicted high environmental prevalence for these novel taxa, particularly within soil habitats. Isolates DC-1.1T, Qj1B1, and DC-1.5T effectively nodulated Lotus burttii and significantly promoted plant growth, whereas Qj2B2 neither nodulated nor enhanced growth. Comparative genomic analysis revealed that the nodulating isolates harbour symbiotic genes (nod, fix, and nif) on symbiotic plasmids, a rare feature in Mesorhizobium strains, whereas Qj2B2 lacks essential nod and nif genes. Consistent with these genomic features, symbiotaxonomic analysis assigned the nodulating isolates to symbiovar loti. These results highlight the potential of these isolates as models for comparative analyses of symbiotic plasmid evolution and horizontal gene transfer.
This study formulated alginate microbeads incorporating a date seed polysaccharide-stabilized pickering emulsion (Alg:PSE) aimed at enhancing probiotic survival and regulating gut microbiota in a strain-dependent manner. Lacticaseibacillus rhamnosus and Bifidobacterium longum subsp. longum strains were individually encapsulated within alginate beads, alginate with the ultrasound-extracted date seed polysaccharide (UPS), and alginate with the UPS-stabilized pickering emulsion (PSE). Among various compositions evaluated herein, Alg:PSE2 was identified as the best one based on the highest encapsulation efficacy and viability rates of L. rhamnosus (AET3) and B. longum (AET8). In comparison with alginate beads, both AET3 and AET8 resulted in denser, harder, less swellable, and heat-stable matrices with smooth and less porous surfaces. Moreover, both microcapsules could significantly enhance the biological activity of bioaccessible fractions following in vitro digestion in terms of their antioxidant capacity and inhibition against α-amylase and α-glucosidase enzymes. Both AET3 and AET8 could stimulate an earlier response of fermentation within 24 h of fecal fermentation with the notable production of propionic and butyric acids. Notably, such treatment caused distinct alterations in microbial community profile by enriching short-chain fatty acids (SCFAs)-associated genera and eliminating the presence of pathogenic genera like Klebsiella. Unbiased metabolomics analysis using LC-MS illustrated distinctive metabolome profiles associated with the predominance of SCFAs, amino acids, and lipids.
Understanding the biomechanical interaction between residual limbs and prosthetic sockets is essential for optimizing prosthesis design and minimizing tissue injury. Existing cadaveric stance simulators have not been specifically designed to examine socket-residual limb biomechanics. This study designed, constructed, and evaluated a cadaveric stance simulator capable of applying physiologically representative compressive forces through residual limbs within sockets. The long-term objective is to utilize dynamic stereo x-ray (DSX) to quantify 3D skin strain at the socket-residual limb interface, for which this simulator serves as a critical foundational step. 
Methods: The simulator combined passive spring compression with active stepper motor control to generate characteristic double-peaked vertical ground reaction force (vGRF) profiles of stance phase. Primary validation was performed under idealized conditions using a rigid prosthesis configuration, with cadaveric testing as a secondary feasibility assessment on two residual limbs with direct force plate measurement. Validation metrics included peak vGRF, peak-to-valley (P2V) ratio, first-to-second peak symmetry (P2P), and loading and unloading rates, with target parameters derived from five participants with transtibial limb loss. Non-parametric analyses compared simulator outputs with control data (α = 0.05). Spring compression-based force estimates were validated against force plate measurements using root mean square (RMS) error analysis. 
Results: Idealized trials reproduced physiological vGRF profiles with no significant differences across validation metrics, confirming accurate stance phase loading reproduction as the primary outcome. Exploratory cadaveric comparisons demonstrated lower peak vGRF, P2V ratio, and unloading rates relative to control participants, attributable to intentional force reductions to preserve tissue integrity. Spring-based force estimates closely approximated force plate measurements.
Conclusion: The stance simulator provided a repeatable, adaptable platform for physiologically representative residual limb loading. Its compatibility with DSX imaging establishes a foundation for quantifying 3D skin strain at the socket-residual limb interface, supporting the improved socket design and outcomes for individuals with lower limb loss.