Search Results (58,188)

Background: Advanced liver fibrosis (LF) is a major determinant of prognosis across chronic liver diseases. Current biomarkers are often etiology-specific and lack cross-cohort robustness. Shared molecular drivers across etiologies remain incompletely defined, and effective anti-fibrotic therapies are limited. Methods: We developed a multi-algorithm consensus machine-learning framework to derive a robust LF progression signature. In the training non-alcoholic fatty liver disease (NAFLD) cohort GSE213621 (n = 368), samples were formulated as a binary classification task (mild fibrosis, F0–F2; advanced fibrosis, F3–F4). Candidate genes were screened in parallel using Boruta, Least Absolute Shrinkage and Selection Operator (LASSO), random forest, and eXtreme Gradient Boosting (XGBoost). Genes selected by at least two algorithms were defined as a high-consensus pool, and genes consistently selected by all four algorithms were prioritized to construct a core signature. Model performance was evaluated by stratified cross-validation in the training cohort and externally validated in four independent cohorts of different etiologies (GSE49541, GSE84044, GSE130970, and GSE276114). Cellular sources of signature genes were characterized using single-cell RNA sequencing (scRNA-seq) datasets GSE136103 (human) and GSE172492 (mouse). For therapeutic discovery, the high-consensus expression profile was queried against the Connectivity Map (CMap) to prioritize compounds predicted to reverse the fibrotic transcriptional program. Withaferin A (WFA) was selected for experimental validation in a carbon tetrachloride (CCl₄)-induced mouse LF model and in the transforming growth factor-β1 (TGF-β1)-stimulated human hepatic stellate cell line LX-2. Bulk liver RNA-seq profiling was performed to interrogate WFA-associated molecular changes in vivo. Results: We identified a six-gene signature (CLEC4M, COL25A1, ITGBL1, NALCN, PAPPA, and PEG3) that discriminated advanced from mild fibrosis, achieving a mean AUC of 0.890 in internal cross-validation and an average AUC of 0.864 across external validation cohorts. scRNA-seq analysis revealed cell-type-specific expression with prominent enrichment in fibroblast populations. In vivo, WFA markedly attenuated CCl₄-induced fibrosis (p < 0.05) and reversed 1314 fibrosis-associated differentially expressed genes (adjusted p < 0.05), which were enriched in fatty acid metabolism and PPAR signaling, as well as extracellular matrix (ECM)–receptor interaction and focal adhesion (adjusted p < 0.05). In vitro, WFA suppressed TGF-β1-induced LX-2 activation, reducing α-SMA and Fibronectin expression (p < 0.05). Conclusions: We report a six-gene signature that robustly predicts advanced LF across etiologies, define its cellular context using single-cell atlases, and validate the anti-fibrotic activity of WFA in both in vivo and in vitro models. Bulk liver RNA-seq and cellular evidence further suggest that WFA-associated effects are linked to lipid metabolic programs, ECM remodeling, and attenuation of hepatic stellate cell activation. Full article

Background/Objectives: Ejiao, a macromolecular protein complex derived from donkey hide, is a traditional Chinese medicine with clinically demonstrated efficacy against ulcerative colitis (UC). Due to its large molecular size and poor absorbability, its therapeutic effects are presumed to depend on gut microbiota. We hypothesized that specific gut bacteria capable of degrading Ejiao might also mediate its biological functions. Methods: To test this hypothesis, a systematic investigation was conducted by integrating culturomics, proteomics, metabolomics, 16S rRNA gene amplicon high-throughput sequencing, and animal disease models. Results: A total of 134 human gut bacterial strains capable of utilizing Ejiao as a nutrient source were isolated. Among them, Ligilactobacillus salivarius D3-8 exhibited the strongest growth in Ejiao-based medium. Genomic analysis identified 63 protease/peptidase genes, and peptidomic profiling confirmed its degradation activity, which released 50 novel peptides. Notably, administration of L. salivarius D3-8 alone significantly alleviated dextran sodium sulfate (DSS)-induced colitis, concurrently increasing the abundance of beneficial bacterium Dubosiella newyorkensis and elevating the anti-inflammatory metabolite indole-3-carbinol via upregulated tryptophan metabolism. Conclusions: Our findings identify L. salivarius D3-8 as both a dedicated Ejiao-degrader and a protective probiotic against colitis. This work provides direct evidence that gut bacteria can utilize Ejiao and proposes a potential novel mechanistic framework in which the biological effects of Ejiao may be mediated through its interaction with specific, functionally potent degraders such as L. salivarius D3-8. Full article

Vegetation in desert ecosystems is strongly affected by seasonal climatic fluctuations and soil physical and chemical properties. Rawdat Nourah is a natural watershed depression within the King Abdulaziz Royal Reserve in Saudi Arabia. It is colonized by grasses, herbs, and shrubs. Climatic variability and soil heterogeneity are influencing the vegetation dynamics and regeneration patterns in this ecosystem. Based on the literature review, no previous study analyzed and determined either the vegetation composition or the soil seed-bank of Rawdat Nourah. So, the general objective of this study is to examine the vegetation composition and its relationships with soil physicochemical properties and soil seed-bank composition across Rawdat Nourah across different seasons. Floristic analyses, vegetation composition, soil properties, and soil seed-bank were performed within two seasons (winter–spring and summer–fall seasons) of 2023–2024. The obtained data were analyzed using multivariate and statistical approaches. Six plant associations were identified: winter–spring (WVG I: Zilla spinosa–Malva parviflora; WVG II: Rhazya stricta–Zilla spinosa; WVG III: Cynodon dactylon–Convolvulus pilosellifolius) and summer–fall (SVG I: Calotropis procera–Pulicaria undulata; SVG II: Cynodon dactylon–Zilla spinosa; SVG III: Rhazya stricta–Schismus arabicus). Species richness was higher in winter–spring (2.4 species stand⁻¹) than in summer–fall (1.66 species stand⁻¹), while the seed-bank densities were 633.9 and 575.1 seeds m⁻², respectively. Vegetation responded strongly to marked seasonal contrasts in temperature and moisture (~15 °C, 11 mm vs. ~36 °C, 3 mm). Moderate human activity enhanced vegetation cover, whereas prolonged grazing exclusion reduced diversity through the dominance of a few species. The response of vegetation structure and species richness to climatic factors varies greatly depending on the increase in water availability, and moisture content during the mild weather Winter–Spring season (mean temperature is 15 °C and rainfall is 11 mm), compared to the Summer–Autumn season (mean temperature is 36 °C and rainfall is 3 mm). The richness and cover of the plants were generally affected by human activity, where long-term grazing will reduce species richness and increase competition between species, making one or two species dominant. Although above-ground vegetation exhibited clear seasonal and spatial shifts in species composition and abundance, these changes were not reflected in the soil seed-bank. This relation suggests that above-ground communities and seed-banks are regulated by different ecological processes under arid conditions. The data of the present study showed low correlation between the current vegetation and the soil seed bank, which reflects a degradation in this region. Therefore, these findings suggest that sustained protection of the King Abdulaziz Royal Reserve is essential for enhancing seed-bank persistence, vegetation recovery, and ecosystem resilience under arid conditions. Full article

Background/Objectives: Humans and other vertebrates contain two estrogen receptors (ERs), ERa and ERb. Among cartilaginous fish (sharks, rays, skates), which are chondrichthyans that evolved about 425 million years ago, only activation by steroids of ERb orthologs has been characterized. To remedy this gap in understanding estrogen signaling in chondrichthyans, we studied estrogen activation of orthologs of human ERa and ERb from elephant shark (Callorhynchus milii). Methods/Results: Unexpectedly, we found that C. milii contained three estrogen-responsive ERa genes: ERa1 (596 amino acids), ERa2 (600 amino acids), and ERa3 (599 amino acids) with strong sequence similarity to each other. We also found an estrogen-unresponsive gene, ERa4 (561 amino acids), with a 39 amino acid deletion in the DNA-binding domain. An estrogen-responsive ERb ortholog (580 amino acids) was also present in C. milii. The three active C. milii ERas are of similar length to human ERa (595 amino acids); however, C. milii ERb is longer than human ERb (530 amino acids). We studied transcriptional activation of ERa and ERb by estradiol (E2), the main reproductive estrogen in humans. We also studied estrone (E1), the main postmenopausal estrogen, and estriol (E3), which is synthesized during pregnancy. We determined the half-maximal response (EC50) and fold-activation to E2, E1, and E3 of C. milii ERa1, ERa2, ERa3, and ERb. Among these estrogens, E2 had the lowest EC50 for all four ERs. Fold-activation by E2 and E3 was similar for ERa1, ERa2, ERa3, and ERb. Conclusions: Overall, estrogen activation of C. milii ERa and ERb was similar to that for human ERa and ERb, indicating substantial conservation of the vertebrate ER during the 425 million years since the divergence of cartilaginous fish and humans from a common ancestor. Full article

The growth of human activity in cities is a key factor in the degradation of air quality. Numerous studies have demonstrated the link between air quality and the existence of dangerous and chronic diseases that are extremely costly for individuals and society. This study presents an analytical framework that compares fixed and mobile air-quality monitoring approaches in cities with limited resources, using Mohammedia city, Morocco, as an example. The framework centers on mobile monitoring units mounted on vehicles and equipped with affordable sensors, GPS technology, and wireless communication systems to track important pollutants, including fine particulate matter (PM_2.5 and PM₁₀) and harmful gaseous compounds (NO₂, SO₂, CO, O₃). The evaluation relies on scenario-based modeling, performance data from existing literature, and calculations of costs throughout the system’s lifetime. To enhance measurement reliability, the researchers developed a correction system that addresses measurement errors caused by temperature, humidity, vehicle speed, vibrations, traffic-related interference, operational interruptions, and communication limitations. The findings indicate that fixed monitoring stations deliver superior measurement precision, with estimated uncertainty ranging from ±1.2–2.5%, though their coverage area is restricted to 0.534 km² (representing 1.6% of Mohammedia). In comparison, the suggested mobile setup could potentially monitor 9.8 km², covering approximately 30% of the city, while decreasing infrastructure needs and setup time (2–4 h compared to 2–4 weeks). Over 10 years, the total cost is EUR 252,000 for mobile monitoring, compared with EUR 3.6 million for a network of 20 fixed stations. These results demonstrate that corrected mobile monitoring systems offer significant promise as an economical and sustainable approach for managing urban environmental conditions. Full article

Wetlands play a vital role in the global carbon cycle and serve as critical carbon sink systems. However, increasing human disturbances and land-use changes have led to widespread wetland degradation, severely weakening their carbon sequestration capacity. This study investigated the Huixian Wetland in the Li River Basin of Southwest China to examine the impacts of wetland degradation on soil physicochemical properties, organic carbon fractions, and carbon fluxes. Based on vegetation and environmental conditions, the wetland was classified into four degradation gradients: non-degraded (ND), slightly degraded (SD), moderately degraded (MD), and heavily degraded (HD), and their spatial differences were systematically analyzed. The results showed that with increasing degradation, soil moisture, total nitrogen, and total phosphorus significantly decreased, whereas soil bulk density and electrical conductivity exhibited an increasing trend. Total organic carbon and active organic carbon fractions, including readily oxidizable organic carbon, light fraction organic carbon, microbial biomass carbon, and dissolved organic carbon, exhibited a pronounced decreasing trend along the degradation gradient, with the decline being most evident in the HD area. Among the labile carbon fractions, microbial biomass carbon (MBC) and light fraction organic carbon (LFOC) exhibited the most drastic declines in heavily degraded areas, indicating their high sensitivity as early warning indicators of wetland degradation. Observations of CO₂ fluxes revealed that from April to September, the net ecosystem exchange (NEE) was negative across all areas, indicating that the wetland functioned as a carbon sink overall. However, NEE values increased with higher degradation levels, suggesting a progressive decline in the carbon sequestration capacity of the wetland; ecosystem respiration (ER) peaked in July and increased with the degree of degradation. The findings indicate that wetland degradation leads to soil environment deterioration, reduction in organic carbon storage, and enhanced CO₂ emissions, ultimately weakening its carbon sink function. To enhance carbon sequestration capacity and maintain ecological functions, sustainable management strategies such as hydrological restoration and vegetation reconstruction are recommended. This study provides a scientific basis for wetland ecological conservation and carbon management in the context of climate change. Full article

Carnosine (or β-alanyl-L-histidine) is an endogenous compound playing very important roles in human organisms as antiglycation and antioxidant agents, and, in addition, helping to mitigate illnesses such as cancer and neurodegenerative diseases. Aiming to explore the chelating ability of carnosine, based on its coordinating possibilities, we started to investigate the metal complexes of essential copper(II), zinc(II), and iron(II) ions coordinated to this dipeptide. Different compounds were isolated in the solid state by adding stoichiometric amounts of metal salts to carnosine at controlled pH or under a controlled atmosphere, with the formation of mono-, bi- and polynuclear species. These complexes were subsequently characterized mainly by spectroscopic techniques (UV–Vis, IR, EPR), in addition to elemental analysis. A binuclear species was isolated with copper(II) and had its structure determined by X-ray diffraction, improving previously reported data in the literature. Two insoluble correlated trinuclear species were isolated with zinc(II) ions, using perchlorate or chloride as counter-ions. In the case of iron, a mononuclear species was verified with Fe(II) ions, obtained under an inert atmosphere. Further, the antioxidant properties of free carnosine and the copper–carnosine complex were verified by their scavenging activity toward the ABTS^•+ radical, using Trolox as a reference, showing significant activity. The carnosine–metal complexes were also tested as potential antineoplastic agents, in comparison to the free ligand, after 24 h of incubation at 37 °C, using malignant HeLa, SKMEL 28 and SKMEL 147, and non-tumor fibroblast cells. Results indicated neglected or poor anti-proliferative properties of these metal complexes, when compared to other similar compounds described in the literature. Full article

Intervertebral disc degeneration is characterized by inflammation, extracellular matrix breakdown, and neurovascular ingrowth, processes that contribute to discogenic, chronic back pain. The transient receptor potential canonical 6 (TRPC6) channel is a calcium-permeable ion channel implicated in inflammation and pain signaling in multiple tissues; however, its functional role in human disc cells remain unknown. Here, we investigated the expression, activation, and downstream consequences of TRPC6 activation using Hyp9, a pharmacological activator of TRPC6. TRPC6 transcripts were consistently detected across all donors examined (n = 17). Functional TRPC6 activation induced a rapid, dose-dependent calcium (Ca²⁺) influx across 0.5–100 µM Hyp9. TRPC6 activation did not reduce metabolic activity or increase cytotoxicity at concentrations commonly used for in vitro TRPC6 activation. Mechanistically, TRPC6 activation induced mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB) pathways, as demonstrated by increased phosphorylation of p38 and extracellular signal-regulated kinase (ERK), degradation of the inhibitor of κB-alpha (IκB-α), and increased nuclear translocation of the NF-κB p65 subunit. Downstream of these early signaling events, TRPC6 activation elicited a robust inflammatory and catabolic response with upregulation of IL-6, IL-8, COX-2, MMP-1, MMP-3, NGF, and VEGF, with corresponding increases in protein secretion. These findings identify TRPC6 as an important signaling node linking calcium influx to inflammatory, catabolic, and neuro- and angiogenesis-associated pathways in disc cells, highlighting TRPC6 as a potential therapeutic target in degenerative disc disease. Full article

Chronic diabetic wounds are challenging to treat due to persistent inflammation, oxidative stress, impaired angiogenesis, and dysregulated matrix remodelling. Hydrogen sulphide (H₂S) has emerged as a therapeutic mediator with antioxidant, anti-inflammatory, and pro-angiogenic properties; however, its clinical translation is limited by volatility and a short biological half-life. Controlled delivery systems, such as hydrogels, are therefore required to harness its potential. This study aimed to develop and evaluate a sodium 2-acrylamido-2-methylpropane sulfonate (Na-AMPS)-based adhesive hydrogel incorporating AP39, a mitochondria-targeted H₂S donor, for sustained localised delivery and promotion of wound healing. Hydrogel formulations were characterised for rheological behaviour, adhesion, swelling, and AP39 release. Cytocompatibility was assessed in human umbilical vein endothelial cells (HUVECs); human dermal fibroblasts, adult (HDFa); and keratinocytes. Anti-inflammatory, antioxidant, and matrix-modulatory effects were evaluated via interleukin-6 and 8 (IL-6/IL-8) secretion, reactive oxygen species (ROS) levels, mitochondrial membrane potential, matrix metalloproteinase-9 (MMP-9), and transforming growth factor-beta (TGF-β). Functional wound healing activity was assessed using tube formation and scratch assays in endothelial cells. AP39-loaded hydrogels exhibited predominantly elastic, shear-thinning behaviour, strong adhesion, rapid hydration, and sustained release of AP39 (11.63 ± 1.20% over 24 h). Across all cell types, 500 nM concentrations of AP39 were well tolerated. In diabetic-like stress conditions, AP39 significantly decreased ROS in HUVECs (50122 ± 5999 to 33,087 ± 1865 AU; p < 0.0001) and HDFa cells (41,367 ± 4225 to 29,813 ± 2406 AU; p < 0.0001). AP39 improved mitochondrial membrane potential in both cell types (p < 0.01–0.001) and decreased pro-inflammatory cytokines. IL-6 decreased in HUVECs (96.05 ± 4.22 pg/mL to 60.99 ± 4.21 pg/mL; p < 0.0001) and HDFa cells (77.54 ± 8.94 pg/mL to 52.25 ± 6.78 pg/mL; p < 0.001), whilst in HDFa cells, MMP-9 was reduced (419.4 ± 25.51 pg/mL to 174 ± 15.1 pg/mL; p < 0.0001). Finally, wound closure was enhanced in HUVECs. The AP39-loaded Na-AMPS hydrogel represents a multifunctional wound dressing capable of controlled H₂S delivery, mechanical stability, and biological activity to support tissue repair in diabetic wound environments. These results highlight this gel’s therapeutic potential for diabetic wound treatment. Full article

Although HyFlex teaching has been studied for decades and has become part of the teaching norm since the 2020 pandemic, studies have generally not investigated the learning experiences of students with disabilities in HyFlex classrooms. This study compared the basic psychological needs (BPN) and engagement of undergraduate students who did (SwA) and did not (SwoA) request academic disability accommodations in an introductory, active learning, human-centered design thinking course, a core component of engineering technology education. Data were collected from 3748 primarily first-year undergraduate engineering technology students between fall 2021 and spring 2024, 126 of whom requested disability accommodation through the disability office. The data sources consisted of an end-of-course survey, in which students reported their basic psychological satisfaction level on a Likert scale and described their BPN experiences and engagement in response to open-ended survey questions. As a novel contribution, this study integrates the descriptive analysis of Likert-scale measures with textual- and word-level sentiment analysis, advancing conceptual understanding of reported BPN satisfaction and engagement and revealing divergent patterns across analytic approaches. While the SwA group reported lower scores across all BPN constructs compared to their counterparts, the highest number of them provided positive feedback statements across all BPN domains. Conversely, the SwoA group reported higher BPN scores across all constructs, yet the highest number of them used negative sentiments in their responses across all BPN constructs. The majority of SwA provided positive feedback on autonomy satisfaction, while the majority of SwoA’s positive feedback was on relatedness to the instructor. Future directions for advancing engineering technology education and disability data collection in higher education are provided. Full article

Background/Objectives: The growing shortage of organs for transplantation requires optimized preservation techniques. Normothermic (NMP) and Subnormothermic Machine Perfusion (SMP) allow for the assessment of organ viability prior to transplantation and enable targeted therapeutic interventions while maintaining a metabolically active state in contrast to hypothermic settings. Methods: In this study, the synthetic perfusion solution “Omnisol” was used in a 6 h ex vivo setting with porcine kidneys (n = 6 NMP; n = 6 SMP). Perfusion parameters (arterial flow, intrarenal resistance and urinary flow), renal function (excretory and filtration performance), renal injury (cellular and circulating biomarkers) and tissue and perfusate oxygenation were assessed. Results: NMP resulted in better arterial flow and lower intrarenal resistance during the first 3 h of perfusion, while SMP surpassed NMP from 3 to 6 h. Renal injury biomarkers increased in the NMP group after 3 h, while no increase was detectable in the SMP group. Omnisol fully met the oxygen requirements of the kidneys in both groups, despite being fully synthetic. Conclusions: Both NMP and SMP offer distinct advantages for kidney preservation, and the synthetic perfusate Omnisol appears to be feasible for both methods. In this experimental setting, the data indicate that NMP of porcine kidneys was associated with favorable functional parameters during the early phase of perfusion, whereas SMP showed comparatively stable parameters at later time points. These findings should primarily be considered exploratory observations and require validation in future studies, especially for the translation into a clinical scenario using human kidneys. Full article

Three new Mn(II) complexes with imidazo[1,2-a]pyridine derivatives were synthesized and structurally characterized in a solid state by single crystal X-ray diffraction, FT-IR and Raman spectroscopy, and thermal analyses. The investigated compounds include [Mn(3-Climpy)₂Cl₂(MeOH)₂] (1), [Mn(3-Brimpy)₂Cl₂(MeOH)₂] (2), and a rare double chloro-bridged coordination polymer [Mn(impy)₂Cl₂]_n (3). Spectroscopic studies were used to assess their potential stability in DMEM (Dulbecco’s Modified Eagle Medium), and encapsulation in Pluronic P-123 micelles improved their solubility in aqueous solution, as well as cellular uptake and selectivity. Biological evaluation revealed negligible cytotoxicity against most cancer and control cell lines, but unexpectedly high activity against pancreatic adenocarcinoma (PANC-1), exceeding that of cisplatin. Complex 2, bearing a bromine substituent in the imidazole ring, showed the strongest effects, correlating with enhanced intracellular accumulation, reactive oxygen species (ROS) generation, and mitochondrial membrane potential disruption. Molecular docking and protein binding assays demonstrated moderate affinity toward human serum albumin (HSA) and transferrin, whereas DNA interaction was weak and non-damaging. These results highlight the structure–activity relationship of Mn(II) imidazo[1,2-a]pyridine complexes and support their potential as targeted redox-active agents against pancreatic cancer, with polymeric encapsulation providing an effective strategy to enhance biological performance. Full article

The essential reaction of CO₂ hydration, fundamental to all living organisms, is facilitated by the enzyme carbonic anhydrase (CA, EC 4.2.1.1). This enzyme plays a crucial role in regulating various physiological and pathological processes. A series of heterocyclic benzenesulfonamide derivatives (19 compounds) were evaluated as possible inhibitors of human CAs. Their inhibitory properties were tested against several isoforms such as the cytosolic hCA I and hCA II, as well as the transmembrane isoforms hCA IV, hCA IX and hCA XII. The tested molecules demonstrated notable inhibitory potential, particularly toward hCA II and hCA IV, where five and four compounds, respectively, exhibited greater potency than the reference inhibitor, acetazolamide. Molecular docking simulations were further performed to elucidate the binding interactions of the most active compounds with the human CA II, IV IX and XII isoforms Full article

Background: Antimicrobial resistance (AMR) is a global healthcare threat. Traditional largely non-selective antibiotics produce side effects due to the natural host microbiome being modified creating a loss in homeostasis. In women, AMR is a cause of acute generational impact. For example, bacterial vaginosis (BV), the most common gynecological infection in reproductive-age women, is a serious public health concern due to its high rates of recurrence, secondary infections, and reproductive issues; and two currently prescribed antibiotics for BV do not fully resolve the symptoms. Objective: The strong need for innovative, potent, safe, and selective therapeutics has prompted a search for such bioactive molecules in milk. Resulting from 200 million years of evolutionary pressure, mammalian lactation not only nourishes infants, but it has also been under relentless Darwinian selective pressure to provide protection from a variety of infections. Methods: Computationally designed human-milk-inspired peptides (AMPs) were tested in standard microbicidal assays for activity against BV pathogens, and evaluated for stability and safety. Results: Several AMPs are bactericidal towards Gardnerella vaginalis, a major BV-associated pathogen, and other BV-associated pathogens. Some novel AMPs do not impact the viability of key lactobacilli linked to a healthy vaginal microbiome. These stable, membrane-acting cationic AMPs reduce inflammation during an infection assay and are safe in EpiVag organoid tissues. Conclusions: AMPs can address concerns like non-selectivity and antibiotic resistance—thereby addressing AMR. Lead AMPs from this study offer a promising solution for the development of novel therapeutics for the treatment of BV, which may reduce the burden of AMR. Full article

Triple-negative breast cancer (TNBC) remains one of the most aggressive breast cancer subtypes and is associated with limited therapeutic options, underscoring the urgent need for novel treatment strategies. In this study, a library of seventeen 1,3,5-triazine derivatives potentially targeting TNBC was developed using an activity-based approach. Compounds were synthesized via an ultrasound-assisted protocol, providing an efficient and environmentally friendly methodology. The synthesized library was evaluated in vitro against the human TNBC cell lines MDA-MB-468, MDA-MB-231, and Hs578T, as well as the non-tumorigenic epithelial cell line MCF10A. Compounds 9 and 17 exhibited the most promising antiproliferative activity against TNBC cell lines (MDA-MB-468: IC₅₀ = 36.62 µM for 9 and 38.29 µM for 17; MDA-MB-231: IC₅₀ = 37.32 µM for 9 and 32.86 µM for 17; Hs578T: IC₅₀ = 57.26 µM for 9 and 34.87 µM for 17), while maintaining acceptable selectivity toward non-cancerous cells. The lead compounds were further assessed in vivo using a Danio rerio model to evaluate general toxicity and cardiotoxicity. In addition, ADME parameters were predicted for all compounds using biomimetic chromatography. Overall, compounds 9 and 17 emerged as promising small-molecule candidates for TNBC treatment, requiring further toxicological evaluation in more human-relevant in vivo models. Full article