Inorganic carbon use in cellular metabolism is ubiquitous in all living organisms. The assimilation of inorganic carbon into biomass via heterotrophic inorganic carbon fixation (HICF) is estimated to contribute significantly to global carbon fixation, yet the controls on this process in strictly heterotrophic microorganisms remain poorly understood. To identify potential drivers of HICF, we cultivated well-characterized heterotrophic Gammaproteobacteria (Pseudomonas putida and Pseudoalteromonas carrageenovora) at two temperatures (optimal vs colder) and resource treatments (high vs low organic carbon), all of which were amended with NaH[13]CO3. We performed bulk and single-cell stable carbon isotope probing (SIP) analysis to detect and quantify HICF in each condition, and assessed other measures of metabolic state, including C/N ratio, natural abundance nitrogen isotope ratios, and cell size. We observed HICF in all treatments, with the percent of biomass originating from [13]C-dissolved inorganic carbon (DIC) for P. putida (0.006% to 0.029%) and P. carrageenovora (0.18% to 1.1%) on the lower end of estimates for the contribution of anaplerotic fixation to heterotrophic biomass (1% to 8%). HICF was generally highest during exponential phase, at the optimal growth temperature, and under low carbon conditions, and these trends were also observed at the single-cell level. Overall, temperature and resource availability appear to influence HICF rates. These findings suggest that HICF may be more relevant under environmental conditions where DIC and nitrogen are not limiting, and the flux of digestible organic carbon is limited or intermittent. Environmental applications of [13]C-DIC SIP in such systems may record heterotrophic contributions to community carbon fixation.IMPORTANCEHeterotrophic microorganisms are abundant across ecosystems and play a central role in environmental carbon cycling by storing, transforming, and decomposing organic matter. However, heterotrophs can also incorporate inorganic carbon into biomass, but this is rarely considered in models of the carbon cycle. Our experiments on heterotrophic bacteria from soil (Pseudomonas putida) and marine environments (Pseudoalteromonas carrageenovora) show that inorganic carbon uptake is impacted by both resource availability and temperature and may be undetectable without stable isotope probing. By integrating single-cell techniques, our study measures variation in inorganic carbon uptake among individual cells across growth phases and conditions. Our study highlights heterotrophic inorganic carbon fixation as a variable source of [13]C-dissolved inorganic carbon incorporation into microbial biomass, underscoring the importance of considering this process in the design and interpretation of environmental stable isotope probing studies.

Lung injury risk from high-velocity blunt impacts is thought to depend on lung pressure. Risk is typically assessed with finite element models (FEMs), but models do not typically account for lung pressure, nor do they account for lung viscoelasticity. The objective of this study is to translate and then validate a set of lung material models that consider both tissue viscoelasticity and pressure in silico. This was completed using one set of quasi-static simulations for translation and one set of dynamic simulations for validation to compare to experimental forces, where CORA was utilized. Digital image correlation (DIC) data of strain and displacement magnitude from the quasi-static experiments were also used to ensure proper material model translation. Three simulations were run for each loading case, where the only difference was the lung material properties to account for pressure (0, 4, and 10 cmH2O). The force-time responses for both simulations showed good agreement with experimental data with a range of 0.77-0.93 CORA scores for the dynamic loading, and a range of 0.84-0.97 CORA scores for the quasi-static simulations, with all simulations falling under "good" or "excellent" categories. The DIC-equivalent data for the quasi-static simulations were determined to be dependent on mesh size. Overall, the quasi-static simulations showed the material was translated into LS-Dyna well, and each of the lung material models were able to reproduce the global force-time responses well in the dynamic simulation set.

Brewer's spent grain (BSG) is an abundant agro-industrial by-product with potential for manufacturing sustainable particleboards, although its relatively low cellulose content limits the mechanical performance of the resulting boards. In this study, particleboards were produced from BSG using a soybean protein concentrate (SPC)-based adhesive at three adhesive contents. In addition, rosin surface coating and wood veneer reinforcement were investigated as strategies to improve the performance of the boards. Physical and mechanical properties were evaluated according to American and European standards, while X-ray micro-computed tomography was used to analyze the structure of the panels. Increasing the SPC content significantly improved all mechanical and physical properties. Rosin coating further reduced water absorption and moisture diffusivity while improving the modulus of rupture, modulus of elasticity, and internal bond strength. X-ray micro-CT analysis revealed local density increases associated with rosin penetration into the board structure. The greatest improvement was achieved with wood veneer reinforcement particleboards, satisfying the H1-ANSI A208.1 and P2-EN312 minimum flexural requirements. Moreover, Digital Image Correlation (DIC) revealed that wood veneer reinforcement significantly altered the deformation and failure mechanisms. These results demonstrate that simple surface reinforcement strategies enable the production of bio-based particleboards with significantly improved performance.

This study investigates the mechanical properties and strength formation mechanism of loess solidified with a ternary blend of fly ash, lithium slag, and magnesium slag. The mix proportion was optimized using response surface methodology with a Box-Behnken design. Unconfined compressive strength (UCS) tests, digital image correlation (DIC), X-ray diffraction (XRD), and scanning electron microscopy (SEM) were employed for evaluation. The optimal 7-day mix (15.926 wt.% fly ash, 10.158 wt.% lithium slag, and 6.427 wt.% magnesium slag) achieved a UCS of 0.974 MPa, while the 28-day optimum (16.064 wt.% fly ash, 10 wt.% lithium slag, and 2 wt.% magnesium slag) yielded 1.834 MPa. Strength development followed a quadratic nonlinear model at early age, shifting to a linear superposition model at 28 days. XRD and SEM revealed that strength enhancement originates from synergistic pozzolanic and hydration reactions under alkaline activation, producing C-S-H gel and ettringite (AFt) that fill pores and cement soil particles. The ternary system demonstrates effective utilization of industrial solid wastes for loess stabilization.

High-fidelity finite-element (FE) simulation of nonlinear full-field responses in AlSi10Mg tensile specimens is computationally expensive, while the available digital image correlation (DIC) data in this study consist only of engineering-strain screenshots rather than exportable numerical fields. This work therefore develops a simulation-to-experiment framework for rapid full-field stress prediction and weakly supervised DIC-informed spatial regularization. Two hundred independent Abaqus/Explicit loading cases generated 5600 graph samples, which were used to train an eight-layer FiLM-conditioned graph neural network for nodal von Mises stress and three-component displacement prediction, together with a decoupled structured force-displacement branch. On independent FE tests, von Mises stress prediction achieved R[2] = 0.9984 with an RMSE of 4.60 MPa, while the dense force-displacement curve achieved R[2] = 0.9957 and RMSE = 59.70 N. Five machine tensile tests gave mean pre-fracture curve NRMSEs of 4.40% for FE versus experiment and 5.31% for GNN versus experiment. On independent specimens L-7 and L-8, the constrained adapter reduced DIC spatial shape loss by 3.21% with a mean absolute stress correction of 0.232 MPa. The framework therefore provides rapid FE-surrogate prediction with conservative experiment-informed spatial regularization rather than experimental stress inversion.

Doxorubicin (Dox)-induced cardiotoxicity (DIC) is a major clinical challenge in cancer therapy. Camel milk exosomes (CMEs) have been applied in anti-tumor treatments as they have a variety of effects, including on inflammation, oxidative stress, metastasis, and apoptosis. However, their role in DIC treatment remains incompletely understood. This research was designed to evaluate the protection provided by CMEs against DIC. The DIC mice were treated with Dox intraperitoneally and divided into a model group and groups treated with different doses of CMEs. Dox-induced H9c2 cell injury was also established and divided into a model group and groups treated with different concentrations of CMEs. The evaluation parameters in vitro included H9c2 cell viability, reactive oxygen species (ROS), mitochondria, and apoptotic cells. The apoptosis and autophagy markers, as well as the nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) pathways, were assessed via Western blotting both in vivo and in vitro. In addition, transcriptome sequencing of cardiac tissue was also applied to investigate the related mechanisms. Our results indicate that CMEs significantly attenuated the cell viability reduction, apoptosis, and ROS production in H9c2 cells caused by Dox. CMEs also regulated autophagy, inhibited apoptosis, and inhibited the NF-κB and MAPK pathways. In conclusion, our findings demonstrate that CMEs exert a cardiac protective effect against DIC by inhibiting apoptosis and regulating autophagy via NF-κB and MAPK signaling pathways.

Doxorubicin (DOX) remains a cornerstone of chemotherapy but is limited by dose-dependent doxorubicin-induced cardiotoxicity (DIC). Recent evidence suggests that mitochondrial dysfunction, particularly oxidative stress and ferroptosis, drives DIC progression. While the flavonoid Calycosin (CAL) is known for its cardioprotective properties, the precise mechanism by which it maintains mitochondrial homeostasis remains elusive. This study aimed to investigate the effects of CAL against DIC and to explore the mechanisms by which CAL regulates mitochondrial integrity and iron homeostasis. Using both in vitro H9c2 cardiomyocytes and in vivo C57BL/6 mice, we demonstrate that CAL significantly restores cardiac contractility and reduces myocardial damage. Transcriptomic analysis indicated that CAL protected mitochondrial function and inhibited ferroptosis. Transmission Electron Microscopy (TEM) showed that CAL preserved mitochondrial structure. Assessments of the lipid peroxidation marker malondialdehyde (MDA) and superoxide dismutase (SOD) confirmed that CAL alleviated oxidative stress, while 8-OHdG staining verified its restoration of mitochondrial DNA integrity. Molecular docking identified the affinity of CAL with NRF2 and NRF1. CAL activated the NRF2/NRF1-TFAM axis both in vivo and in vitro, promoting mitochondrial biogenesis and respiratory function, while regulating iron homeostasis via FTMT and FPN. Knockdown or inhibition of NRF2/NRF1 abolished these protective effects of CAL. Our findings suggest that CAL acts as a dual-regulator of mitochondrial transcriptional machinery and iron metabolism, offering a novel therapeutic strategy to safeguard mitochondria against ferroptosis.

Arthroscopic dorsal capsuloligamentous reconstruction (ADCLR) has gained popularity in addressing symptomatic scapholunate instability without radiographic malalignment (European Wrist Arthroscopy Society [EWAS] grade IIIB/IIIC/IV).

Acid mine drainage (AMD), characterized by low pH, high sulfate concentrations, and toxic heavy metals, poses severe ecological and human health risks. This study compared the performance of a conventional aerated algal-bacterial biofilm reactor (CAABR) and a membrane-aerated algal-bacterial biofilm reactor (MAABR) for AMD treatment. Six reactors with different influent COD concentrations (400 ± 11.41, 1400 ± 19.07, and 2800 ± 18.37 mg/L) were operated for 80 days, and pollutant removal, biofilm properties, and microbial community dynamics were systematically analyzed. Results showed that the MAABRs and CAABRs exhibited broadly comparable sulfate-removal performance, whereas the MAABRs maintained greater aqueous DIC availability and more stable heavy-metal removal under shock loading. Kinetic analysis revealed a transition from algal assimilation (first-order) to SRB-mediated reduction (zero-order) as COD increased, confirming that COD/SO4[2-] ratios regulate pathway dominance. Multiscale biofilm characterization and EPS fluorescence analysis demonstrated that membrane aeration improved CO2/O2 mass transfer, enhanced algal biomass, and maintained higher EPS secretion under stress. High-throughput sequencing revealed the simultaneous enrichment of Pseudomonas and the sulfate-reducing genus Desulfosporosinus, indicating that the MAABR supported the coexistence of functionally distinct microbial populations. Overall, this study demonstrates that membrane aeration enhances algal-bacterial symbiosis, improves pollutant removal efficiency, and strengthens system resilience, providing a promising and sustainable approach for AMD remediation.

Doxorubicin (DOX) has antitumor efficacy, but its clinical application is restricted by multiple side effects, especially cardiotoxicity. Neohesperidin, a flavonoid in citrus fruits, has various health benefits. The aim of this study was to explore the cardioprotective effects and mechanisms of neohesperidin against DOX-induced cardiotoxicity (DIC). The study used DOX-treated mice to evaluate cardiac function and cardiomyocyte injury. In addition, primary cardiomyocytes treated with DOX were used to measure cell viability and death. FIN56, a ferroptosis inhibitor, was used to test the relationship between neohesperidin's beneficial effect and ferroptosis. Mechanistic investigations were carried out to explore the role of N6-methyladenosine (m[6]A) levels, nuclear factor erythroid-2-related factor 2 (Nrf2) expression, and Methyltransferase-like 3 (Mettl3) in the cardioprotective effect of neohesperidin. Overexpression and knockdown of Mettl3 were also performed to confirm its role in the cardioprotective effect. Neohesperidin attenuated cardiac function and cardiomyocyte injury in DOX-treated mice. It alleviated DOX-induced oxidative stress by reducing reactive oxygen species and malondialdehyde levels and increasing superoxide dismutase and glutathione levels, restoring ATP production. In DOX-treated primary cardiomyocytes, neohesperidin increased cell viability and decreased cell death. FIN56 abolished the beneficial effect of neohesperidin. Mechanistically, neohesperidin decreased m[6]A levels, leading to increased Nrf2 expression. Nrf2 activation was related to the inhibition of Mettl3, an m[6]A "writer," which protected cardiomyocytes against DOX-induced ferroptosis. Overexpressing Mettl3 reversed the cardioprotective effects of neohesperidin and diminished its benefits on ferroptosis. These findings suggest that neohesperidin exerts a cardioprotective effect against DIC by inhibiting ferroptosis. This effect is achieved by regulating the Mettl3-m[6]A-Nrf2 signaling pathway, indicating the therapeutic potential of neohesperidin.

Bovine brucellosis is a major zoonotic disease in Pakistan, yet diagnostic tests used in national surveillance have never been validated under field conditions. This cross-sectional study estimated the sensitivity (Se) and specificity (Sp) of three commonly used bovine brucellosis serological tests, viz. Rose Bengal plate test (RBPT), Serum agglutination test (SAT), and indirect ELISA (iELISA), and the true prevalence of the disease in the study area using Bayesian latent class analysis (BLCA). Serum samples from 1000 animals (507 cattle, 493 buffaloes) across four districts (Faisalabad, Jhang, Chiniot and T.T. Singh) were tested in parallel. Since all three assays target antibodies against smooth lipopolysaccharide (S-LPS), full conditional dependence (CD) was modeled, with an acceptable fit attained only under informative priors (posterior predictive p-value = 0.250; DIC = 211.73). RBPT showed the lowest Se (0.705), while SAT (0.938) and iELISA (0.952) performed substantially better. All three tests achieved high Sp (RBPT = 0.977; SAT = 0.993; iELISA = 0.997). True prevalence ranged from 7.5% (Faisalabad) to 25.3% (Jhang). SAT + iELISA in parallel achieved the highest performance index (PI = 1.961; Se = 0.971, Sp = 0.990). Species-level analysis revealed higher SAT sensitivity in buffaloes (0.969) than cattle (0.849), with a notable prevalence divergence in Jhang (cattle = 39.6%; buffaloes = 10.7%). It was concluded that a single diagnostic test alone is unreliable for screening in similar settings. SAT + iELISA in parallel is recommended for field screening and in series for culling decisions. Significant district-level variance in prevalence and species-specific diagnostic performance indicates that uniform provincial strategies may be inappropriate and that control programs should be designed at the district level and calibrated separately for cattle and buffaloes.

Tropical high-mountain lakes represent a biogeochemically distinct but poorly characterized subset of global lake research, with their greenhouse gas (GHG) dynamics remaining largely unexplored. Here, we present the first comprehensive assessment of CO2, CH4 and N2O fluxes, dissolved concentrations, and isotopic signatures in high-mountain Andean lakes (3,100-4,450 m elevation). We combined diel measurements, depth profiling, and stable isotope analyses (δ[13]C-CO2, δ[13]C-DIC, δ[13]C-CH4, and δ[15]N- and δ[18]O-N2O) to characterize GHG sources, production pathways, and environmental drivers across gradients of morphometry, lithology, and human impact. Shallow wetland-connected lakes functioned as strong CO2 (up to 1,639 mg m[-2] d[-1]) and CH4 (up to 604 mg m[-2] d[-1]) sources but persistent N2O sinks (up to -45.4 µg m[-2] d[-1]), while deeper, less connected lakes exhibited lower CO2 and CH4 emissions and slight N2O effluxes. Diel patterns revealed metabolic control over CO2 dynamics, with nighttime fluxes 3.2 times higher than daytime values, and temperature-driven episodic CH4 flux peaks in the shallowest systems. Deep lakes maintained well-oxygenated water columns with respiratory CO2 accumulation at depth, while elevated surface CH4 suggested the presence of lateral littoral inputs and oxic methane paradox production. Isotope signatures indicated that CO2 originated primarily from C3 plant organic matter respiration and bedrock composition (δ[13]C-CO2: -40 ‰ to -16 ‰), CH4 from biogenic methanogenesis with partial oxidation during upward transport, and minimal N2O cycling under nitrogen-limited conditions. Anthropogenic disturbance amplified CH4 emissions by an order of magnitude while enhancing N2O consumption through complete denitrification. Our findings demonstrate that Andean remote oligotrophic lakes are sensitive to even minor catchment disturbance and exhibit distinct GHG dynamics given their unique characteristics, with implications for incorporating high-mountain lakes into global emission inventories.