Search Results (14,052)

Afatinib and osimertinib are current treatment options for non-small cell lung cancer (NSCLC) patients with uncommon epidermal growth factor receptor (EGFR) mutations, although their efficacy is limited. To explore potentially effective drugs for these patients, we evaluated the efficacy of conventional EGFR tyrosine kinase inhibitors (TKIs) and novel third-generation (3G) TKIs using in vitro models. Ba/F3 cells transformed with each of the five most frequent uncommon EGFR mutations, Del18 (delE709_T710insD), E709K, G719A, S768I, and L861Q, were used. The growth inhibitory effects of five novel 3G-TKIs, almonertinib, lazertinib, furmonertinib, rezivertinib, and befotertinib, in addition to currently available TKIs, were evaluated. We also explored for secondary resistant mutations to afatinib or osimertinib and TKIs that can overcome these resistances. Afatinib was active against all uncommon EGFR mutations tested. The 3G-TKIs were all active against the L861Q mutation and were inactive against the S768I mutation. Furmonertinib and befotertinib showed efficacy against exon 18 mutations (Del18, E709K, and G719A). In the acquired resistance models to afatinib or osimertinib, we found T790M or a novel T725M secondary mutation, respectively, both of which could be overcome by lazertinib. However, some afatinib-resistant cells acquired V769L/M secondary mutations that were refractory to all EGFR-TKIs tested. In conclusion, afatinib exhibited broad activity and some 3G-TKIs showed promising efficacy in the front-line setting. Lazertinib is a potential second-line option after acquisition of resistance to afatinib or osimertinib. Full article

Chronic respiratory diseases claim nearly four million lives annually, making them the third leading cause of death worldwide. Extracorporeal membrane oxygenation (ECMO) is often the last line of support for patients with severe lung failure. Still, its performance is limited by an incomplete understanding of gas exchange in hollow fiber membrane (HFM) oxygenators. Computational fluid dynamics (CFD) has become a robust oxygenator design and optimization tool. However, most models oversimplify O₂ and CO₂ transport by ignoring their physiological coupling, instead relying on fixed saturation curves or constant-content assumptions. For the first time, this study introduces a novel physiologically informed CFD model that integrates the Bohr and Haldane effects to capture the coupled transport of oxygen and carbon dioxide as functions of local pH, temperature, and gas partial pressures. The model is validated against in vitro experimental data from the literature and assessed against established CFD models. The proposed CFD model achieved excellent agreement with experiments across blood flow rates (100–500 $\mathrm{mL}/\min$ ), with relative errors below 5% for oxygen and 10–15% for carbon dioxide transfer. These results surpassed the accuracy of all existing CFD approaches, demonstrating that a carefully formulated single-phase model combined with physiologically informed diffusivities can outperform more complex multiphase simulations. This work provides a computationally efficient and physiologically realistic framework for oxygenator optimization, potentially accelerating device development, reducing reliance on costly in vitro testing, and enabling patient-specific simulations. Full article

Background: A novel series of pyrazolo[3,4-d]pyrimidinone derivatives were synthesized, characterized, and examined for their anti-inflammatory effects. Results: The findings indicated that compounds 5d, 5j, 5k, and 5m demonstrated significant anti-inflammatory effects through the selective inhibition of the COX-2 isozyme, with IC₅₀ values ranging from 0.27 to 2.34 μM, compared to celecoxib (IC₅₀ = 0.29 μM). Compound 5k emerged as the most potent, exhibiting a selectivity index (SI) of 95.8 for COX-2 relative to COX-1. In vivo tests additionally validated that compounds 5j and 5k demonstrated significant anti-inflammatory efficacy, exhibiting greater suppression percentages of generated paw edema than indomethacin, comparable to celecoxib, while preserving excellent safety profiles with intact gastric tissue. Mechanistic studies demonstrated that the anti-inflammatory efficacy of the target compounds was associated with a substantial decrease in serum levels of TNF-α and IL-6. Moreover, molecular modeling investigations corroborated the in vitro findings. Compound 5k displayed a binding free energy ΔG of −10.57 kcal/mol, comparable to that of celecoxib, which showed a ΔG of −10.19 kcal/mol. The intensified binding contacts in the COX-2 isozyme indicated the augmented inhibitory efficacy of 5k. Conclusions: Compound 5k exhibited dual activity by inhibiting the COX-2 isozyme and suppressing the pro-inflammatory cytokines TNF-α and IL-6, therefore providing a remarkable anti-inflammatory effect with increased therapeutic potential. Full article

Background/Objectives: IBDs are chronic relapsing inflammatory intestinal disorders whose precise etiology is still only poorly defined: critical for their pathogenesis is the CCL20/CCR6 axis, whose modulation by small molecules may represent an innovative therapeutic approach. The aim of the present work is to test the potential efficacy of two molecules, MR120, a small selective CCR6 antagonist, active in TNBS- and chronic DSS-induced murine models of intestinal inflammation, and its derivative MR452, a well-tolerated agent endowed with improved anti-chemotactic in vitro properties, in the adoptive transfer colitis model. To the best of our knowledge, this is the first attempt to use adoptive transfer colitis to test modulators of the CCL20/CCR6 axis. Methods and Results: The induction of colitis in immunocompromised mice receiving CD4⁺CD25⁻ T cells i.p. resulted in a moderate inflammation and was met with limited protective responses following daily subcutaneous administration of MR120 or MR452 for 8 weeks. Both compounds significantly reduced colonic myeloperoxidase activity, and MR452 also lowered CCL20 levels in the gut, but they failed to prevent the increase in the Disease Activity Index, colon wall thickening, and macroscopic inflammation score. Conclusions: Our findings suggest that, despite the beneficial effects played by MR120 against subacute TNBS- and chronic DSS-induced colitis, the pharmacological targeting of the CCL20/CCR6 axis in the adoptive transfer model has a negligible effect in ameliorating the IBD-like phenotype driven by the altered intestinal immune homeostasis and by the disrupted function of immune-suppressive Treg cells. Full article

In the last decades, extracts of Viscum album L., commonly known as European mistletoe, have attracted increasing interest for their immunomodulatory, anti-inflammatory and antioxidant activities. Nowadays, they are mainly used in complementary cancer treatments. A targeted LC-MRM-MS was selected to determine the chemical composition and the activities of a V. album homeopathic mother tincture (MT#39998). Results showed a complex chemical composition, which was compared with that of other similar extracts. The LC-MRM-MS data were confirmed and complemented by HPLC analysis. Viscotoxins content was evaluated because of their cytotoxicity. MT#39998 was tested for its cytotoxic and antioxidant effect, before and after viscotoxins removal. The composition of MT#39998 in viscotoxins was similar to that of other products already present in the market and its safety was confirmed by estimation of LD₅₀ based on in vitro IC₅₀ values (LD₅₀ was >2 g/kg). The aim of this study is to report a case study on a plant extract. The study was based on the chemical composition, including the metabolome, and on the pharmacological data, including toxicity and antioxidant activities, to validate the current utilization. Full article

Porcine epidemic diarrhea virus (PEDV) is a highly contagious coronavirus that causes severe diarrhea, dehydration, and high mortality in piglets, leading to significant economic losses in the swine industry. The spike (S) protein of PEDV is the primary target for neutralizing antibodies and is critical for vaccine development. In this study, the pUC57-S01 and pUC57-S02 plasmids carrying the codon-optimized truncated S gene sequence were constructed. The mRNA S01 showed higher protein expression in vitro than mRNA S02, as confirmed by Western blotting. The safety and immunogenicity of mRNA S01 were evaluated in animal experiments. The results indicated that the mRNA S01 vaccine was safe for piglets and pregnant sows. Immunogenicity was assessed by a neutralization assay, which revealed that encapsulated mRNA S01 induced high levels of neutralizing antibody titers in pigs. Challenge protection efficiency tests showed that the mRNA S01 vaccine conferred immunity to newborn piglets, protecting them from a homologous PEDV strain challenge. This study provides a foundation for the clinical application of PEDV mRNA vaccines and offers a reference for the development of novel vaccines against PEDV. Full article

With the rapid evolution of nanotechnology, magnetic iron oxide nanoparticles (MNPs)—primarily Fe₃O₄ and γ-Fe₂O₃—have gained prominence in biomedicine. Their extensive specific surface area, tunable surface functionalities, and intrinsic magnetic characteristics render them highly versatile for diverse clinical applications, including tumor visualization through Magnetic Resonance Imaging (MRI), radiolabeling, targeted radiotherapy, hyperthermia, gene transfer, drug delivery, Magnetic Particle Imaging (MPI), magnetic blood filtration and theranostic strategies. Nevertheless, ensuring the biocompatibility and non-toxicity of these nanostructures remains a fundamental prerequisite for their medical implementation. Hence, it is essential to continuously refine our understanding of MNP-related toxicity and pursue comprehensive research on this front. This article consolidates up-to-date insights into the evaluation of MNPs’ toxicological profiles, emphasizing the influence of physicochemical properties such as morphology, surface modifications, and electrostatic characteristics, along with operational factors like dosage and administration routes. Traditional toxicity testing strategies, including in vitro assays as first-line screening tools, together with standard ex vivo and in vivo models, are discussed. Special attention is given to the emerging role of New Approach Methodologies (NAMs), such as organoid formation, 3D bioprinting, in ovo chicken embryo assays, and image cytometry. These techniques offer ethical, human-relevant, and informative alternatives to animal testing, supporting more predictive and translationally relevant toxicity assessment of MNPs. Taken together, the integration of conventional assays with innovative NAMs, alongside careful consideration of physicochemical and operational factors, is essential to translate the laboratory promise of MNPs into safe and clinically effective nanomedicines. Full article

Curcumin is a natural bioactive compound of plant origin, characterised by a wide variety of properties that make it useful in numerous industries. Furthermore, due to its health-promoting properties, such as anti-inflammatory, antioxidant, and antimicrobial effects, it has found applications in medicine and animal husbandry. Unfortunately, curcumin has low bioavailability; its hydrophobic nature means it is poorly absorbed through the gastrointestinal tract, and it is rapidly metabolised in the liver. In recent years, research has been conducted into adding nanoencapsulated active ingredients, such as curcumin, to animal feed. This research aims to improve the bioavailability and stability of these ingredients, extend their shelf life, and enhance their absorption. These effects are expected to improve overall animal health, increase production efficiency, and enhance the quality of animal products. However, a significant challenge remains: the irreversible aggregation and chemical instability of bioactive substances due to the hydrolysis of their polymeric encapsulants, which can lead to toxic effects. This study utilised peripheral whole blood from five Blanc de Termonde rabbits. In vitro cell exposure was conducted using three distinct concentrations of nanoencapsulated curcumin (C1–C3: 10, 5.0, and 2.5 µg/mL) and a control. Cytotoxicity was determined by assessing viability using trypan blue exclusion, the comet assay, and the micronucleus assay. The results indicated that all tested concentrations of nanocurcumin significantly decreased the viability of blood cells to approximately 1–9%. In contrast, the encapsulation matrices themselves were not toxic (results were statistically significant). In the comet assay, the nanocurcumin formulations were toxic at all concentrations, and the results were statistically significant. Following exposure, the micronucleus assay revealed cell damage and a high percentage of apoptotic cells (up to 30% for Cur1 at 10 ug/mL). A significant number of binucleated cells with two micronuclei (BNCs + 2MN) were also observed, again for Cur1. In view of the considerable variation in the results from the individual tests, it is advisable to repeat the research using different matrix forms and concentrations of curcumin. Full article

Skin, the largest organ of the human body, serves as a critical physico-chemical barrier against environmental insults and plays essential roles in hydration, thermoregulation, immune defense, and metabolic functions. Wound healing is a complex, multistage biological process involving hemostasis, inflammation, proliferation, and remodeling. Hydrogels have emerged as a promising class of wound dressings due to their high moisture retention, biocompatibility, and ability to mimic the extracellular matrix, thereby supporting accelerated healing and controlled drug delivery. This review provides a comprehensive overview of current hydrogel types—classified by origin, crosslinking mechanisms, and responsiveness to stimuli—and evaluates their use in experimental research on in vitro, ex vivo, and in vivo wound healing models. Furthermore, clinical applications of hydrogels in wound therapy are discussed. Advances in semisynthetic and stimuli-responsive hydrogels, along with improved testing models, offer enhanced therapeutic potential and underscore the need for continued innovation to optimize wound care outcomes and alleviate healthcare burdens. Full article

Background: Three-dimensional (3D) printing has emerged as a valuable tool in dentistry for producing provisional restorations with high precision and reduced costs. However, the limited mechanical strength of temporary 3D-printed resins remains a clinical concern. This in vitro study aimed to enhance the mechanical properties of a 3D-printed temporary resin by incorporating functionalized niobium (Nb) nanoparticles and to compare its performance with a conventional resin composite and a permanent 3D-printed resin. Methods: Six groups were evaluated: bisacrylic resin (Protemp), resin composite (Z350), temporary 3D resin (Temp 3D), permanent 3D resin (Perm 3D), Temp 3D + 0.05% Nb, and Temp 3D + 0.1% Nb. Niobium oxyhydroxide nanoparticles were synthesized using a hydrothermal method, silanized, and incorporated into the Temp 3D at 0.05% and 0.1% by weight. The tested variables included flexural strength (FS), elastic modulus (EM), surface hardness (SH), and color stability (ΔE). Results: The Z350 resin showed the best mechanical results. The addition of 0.1% Nb nanoparticles significantly improved the FS, EM, and SH of the Temp 3D, reaching values comparable to the Perm 3D (p > 0.05). Color stability remained unaffected across all groups. Conclusions: These findings suggest that Nb reinforcement at a low concentration is a promising strategy for improving the performance of 3D-printed temporary restorations. Full article

Background/Objectives: Composite resins are widely used in restorative dentistry due to their aesthetic properties and ease of handling. Preheating prior to light polymerization has been proposed to improve flowability, degree of conversion, and mechanical properties. This in vitro study aimed to evaluate the effect of preheating on the microhardness of three nanoparticulate composite resins—IPS Empress Direct (Ivoclar), Filtek Z350 XT (3M-ESPE), and Forma (Ultradent)—when cured with a high-power LED light. Methods: Sixty disc-shaped samples (n = 20 per material) were fabricated and divided into preheated and non-preheated groups. After polishing and 24 h storage in distilled water at 37 °C, samples were subjected to Knoop microhardness testing under a 300 g load for 15 s. Statistical analysis was conducted using R software. Results: Preheating produced a significant increase in surface microhardness for IPS Empress Direct (32.8%) and Filtek Z350 XT (5.8%) (p < 0.05 for both), whereas Forma showed no significant change. Conclusions: Under the conditions of this in vitro study, preheating can enhance the mechanical performance of specific composite resins by increasing microhardness; however, the effect is material-dependent. Full article

Given the growing consumer demand for improved quality of life and health-promoting foods, replacing conventional fats in widely consumed products such as bread with oils derived from native Brazilian fruits represents a promising strategy. This study aimed to evaluate the bioactive and technological potential of buriti (Mauritia flexuosa) and acuri (Attalea phalerata) oils, extracted from palm fruits native to the Cerrado and Amazon biomes. Both oils proved to be rich sources of lipophilic bioactives, particularly carotenoids, tocopherols, and phenolic compounds, and exhibited excellent carotenoid bioaccessibility under in vitro digestion, with recovery rates of 74% for acuri oil and 54% for buriti oil. Notably, buriti oil showed a high β-carotene content (1476.5 µg/g). When incorporated into sandwich bread formulations, these oils enhanced antioxidant activity, improved texture, volume, and color, and maintained high sensory acceptance compared to bread made with soybean oil. Sensory evaluation scores averaged above 7 for all tested attributes. These findings underscore the industrial applicability of buriti and acuri oils as functional lipids aligned with sustainable development and nutritional innovation. Full article

The common krait (Bungarus caeruleus) and the endemic Sri Lankan krait (B. ceylonicus) are two species of krait responsible for envenomings in Sri Lanka that result in progressive neuromuscular paralysis. We characterised the in vitro neurotoxicity of B. ceylonicus and B. caeruleus venoms and studied their neutralisation by two commercially available Indian polyvalent antivenoms (i.e., VINS and BHARAT), Thai banded krait antivenom and Australian polyvalent antivenom using the chick biventer cervicis nerve-muscle preparation. Both venoms displayed concentration-dependent neurotoxicity, showing equipotent pre-synaptic neurotoxicity at 0.03 μg/mL. At a higher concentration (1 μg/mL), both venoms showed post-synaptic neurotoxicity, with B. ceylonicus venom being more potent. VINS was unable to neutralise the neurotoxicity of B. ceylonicus venom, but neutralised both pre- and post-synaptic neurotoxicity of B. caeruleus venom. BHARAT neutralised in vitro pre- and post-synaptic activity of both B. ceylonicus and B. caeruleus venoms. Banded krait antivenom and Australian polyvalent antivenoms were unable to fully neutralise the neurotoxicity of either venom at tested concentrations. In conclusion, B. ceylonicus venom shows pre- and post-synaptic neurotoxicity similar to B. caeruleus venom. BHARAT effectively neutralises both pre- and post-synaptic neurotoxicity of B. ceylonicus venom. Both Indian polyvalent antivenoms effectively neutralise neurotoxicity induced by B. caeruleus venom. Full article

Background: Lung adenocarcinoma (LUAD), the most prevalent and malignant form of lung cancer subtypes, is in urgent need of additional therapeutic targets and prognostic indicators. Antioxidant 1 (ATOX1) copper chaperone and RhoA/Rho kinase 1 (ROCK1) are novel anti-tumour targets in cancers. However, their prognostic value and synergistic inhibitory effect remain unclear in LUAD. Methods: We re-analyzed the open-access proteomic landscape study of LUAD in 2019 and investigated the prognostic value of ATOX1/ROCK1 expression patterns. Then we verified it immunohistochemically using an independent cohort from our hospital enrolling 35 patients with TNM stage III/IV LUAD. In vitro, double fluorescence was used to confirm the co-expression and location of ATOX1/ROCK1. The CCK—8 assay and Transwell assay were carried out to assess the changes in proliferation and migration of Lewis lung carcinoma (LLC) cells following treatment with ATOX1/ROCK1 si-RNA or inhibitory drugs. Western blot was used to confirm protein expression after si-RNA transfection. Moreover, ATOX1/ROCK1-targeted drugs’ therapeutic effects were further investigated in the LLC allogeneic transplantation model and MNU-induced tumour model. Results: Firstly, according to the ATOX1/ROCK1 expression pattern derived from proteomic data, double-low expression of ATOX1/ROCK1 indicated a better Disease Free Survival (DFS) (log-rank test p = 0.01) and Overall Survival (OS) (log-rank test p = 8.2 × 10⁻³), whose expression was also correlated with the lower expression of MCM family proteins. Further, we verified this prognostic correlation in our cohort. The IHC-defined ATOX1/ROCK1 low subtype also had the best OS (log-rank test p = 2.4 × 10⁻³). In vitro, double fluorescence confirmed that ATOX1/ROCK1 was highly expressed together in Lewis cells. Co-inhibition of ATOX1 and ROCK1 either by siRNA transfection or inhibitory drugs could lead to a significant decrease in tumour proliferation. Interestingly, transcriptional inhibition of ATOX1 can lead to the up-regulation of ROCK1, while inhibition of ROCK1 resulted in the promotion of ATOX1. Moreover, in the analysis of migration ability, a similar synergistic effect from the co-inhibition of ATOX1/ROCK1 was also observed. Finally, the Lewis and Mnu-induced allogeneic transplantation model also demonstrated a greatly improved therapeutic effect by combining targeting ATOX1 and ROCK1. Conclusions: Collectively, our results suggest that a low expression pattern of ATOX1/ROCK1 can predict better clinical outcomes in LUAD. Combining the inhibition of these two targets can reach a significantly better therapeutic effect than targeting either alone. Full article

Bacteriophage therapy can successfully provide additional treatment to control Salmonella infection, but low gastric pH limits its oral application. The present study aimed to develop an improved encapsulation formulation with enhanced acid protection for oral delivery of Salmonella phages using polymers. This was achieved by encapsulating a phage cocktail containing three different bacteriophages against Salmonella sp. in alginate beads incorporating polyvinyl alcohol (PVA), PVP-K30, and calcium carbonate as viscosity modifiers and acid protection enhancers. Further, the beads were coated with poly-L-lysine to improve the stability and tested for their efficacy for improved phage viability under in vitro acidic conditions for subsequent use in oral delivery. Moist beads were slimy, and semi-dried beads presented a coarse surface as observed using FE-SEM. In vitro studies revealed that the free phage cocktail exhibited complete inactivation when exposed to acidic pH 2.5 after 15 min incubation. In contrast, the encapsulated phage cocktail showed a decrease of only 1.66 log units in viability when incubated for 90 min at pH 2.5. Furthermore, oral delivery of the encapsulated phage cocktail in the poultry model significantly reduced bacterial load in infected birds’ intestines. Full article