Butyrylcholinesterase (BChE) plays a key role in preserving appropriate cholinergic neurotransmission that is essentially altered in the brains of advanced Alzheimer's disease (AD), hence a therapeutic target. This study employed a machine learning (ML) bioactivity predictive model to explore the chemical space of potential BChE inhibitors. A cheminformatics pipeline was explored to create a ML model for BChE inhibition using structural insights complemented with a comprehensive variance importance plot (VIP) and correlation matrix analysis. Specifically, a compiled library of 2179 secondary metabolites (SMs) from 50 Nigerian medicinal plants with reported cholinergic activity was investigated using the ML model. After which, molecular modelling was used to further screen the active SMs (827). The final predicted models demonstrated significant robustness, with a correlation coefficient of 0.8981. Molecular docking investigation of the 827 SMs identified the top five candidates based on their scores: three triterpenoids (adipedatol, lupenone and β-amyrin), one steroid (9 (11)-dehydroergosterol benzoate) and one flavonoid (tiliroside). These leads exhibited favourable ADMET properties and promising safety profiles. Among these five, lupenone (-48.78 kcal/mol), β-amyrin (-49.19 kcal/mol) and adipedatol (-48.87 kcal/mol), from Peltophorum pterocarpum, Bryophyllum pinnatum and Alchornea laxiflora, respectively, were the most promising leads with significant binding free energy compared to decamethonium (-17.64 kcal/mol) and more favourable van der Waals, electrostatics and nonpolar solvation energetics. Furthermore, the binding of these leads resulted in optimised interaction profiles that preserved the structural integrity of the BChE and aligned well with desirable drug-like characteristics. These findings position the leads as promising candidates for therapeutic applications targeting BChE for AD management, subject to further in vitro and in vivo validation investigations.
Antibiotic resistance has emerged as a major global health challenge, particularly in biofilm-forming pathogens that exhibit enhanced tolerance to antimicrobial therapies. K. pneumoniae, a multidrug-resistant pathogen, is a leading cause of hospital-acquired infections, including pneumonia, septicemia, and device-associated infections. Its robust biofilm-forming capacity facilitates immune evasion, restricts antibiotic penetration, and contributes to recurrent infections. Therefore, the identification of agents capable of targeting both planktonic bacterial growth and biofilm architecture is of considerable therapeutic importance. In the present study, the antibacterial, antibiofilm, and antivirulence potential of 6-Aminoflavone was investigated against K. pneumoniae. Antibacterial activity was evaluated using growth inhibition assays, time-kill kinetics, and clonogenic survival analysis. Mechanistic investigations revealed significant intracellular ROS accumulation, increased oxidative stress susceptibility, and induction of apoptosis-like cell death, suggesting ROS-mediated antibacterial activity. In addition to suppressing bacterial proliferation, 6-Aminoflavone exhibited promising antibiofilm efficacy, inhibiting biofilm formation by 86.23% and eradicating 84.77% of established biofilms. These effects were associated with a substantial reduction in cell surface hydrophobicity (∼45.39%), and EPS levels were reduced to ∼2.33% as compared to untreated controls, indicating severe destabilization of the biofilm matrix. Furthermore, eDNA, a key structural component of the biofilm scaffold, exhibited a maximum ∼2.13-fold reduction following treatment. Confocal microscopy confirmed marked disruption and collapse of biofilm architecture. Additionally, quorum sensing (QS)-regulated virulence factors, including urease (∼51.92%), protease (∼55.18%), and lipase (∼44.90%), were significantly attenuated. These findings demonstrate that 6-Aminoflavone exerts antimicrobial activity by inducing oxidative stress-mediated apoptosis-like bacterial death, disrupting biofilm structural components, and suppressing QS-regulated virulence in K. pneumoniae.
Plasmodium falciparum is the most virulent human malaria parasite and is responsible for numerous deaths annually. The increasing resistance of P. falciparum to antimalarial drugs necessitates the development of improved diagnostic tools for timely malaria detection. Malaria biomarkers such as PfHSP70 and PfLDH are highly valuable for malaria detection because they are essential for parasite survival and are consistently expressed during infection. PfHSP70 is associated with the parasite's stress adaptation and proteostasis mechanisms under febrile and drug-induced conditions, while PfLDH plays a central role in glycolytic metabolism and redox balance. Their functional importance, parasite specificity and elevated expression during active infection make these proteins reliable molecular indicators for the sensitive and specific detection of P. falciparum, thereby supporting their potential application in rapid diagnostic and biosensing platforms for malaria surveillance and disease management. In this study, an M13 phage-displayed single-chain variable fragment (scFv) antibody library was used to screen for antibodies against recombinant PfHSP70 and PfLDH. Four rounds of biopanning were conducted to enrich high-affinity binders, followed by ELISA, UV-visible spectroscopy and microscale thermophoresis (MST) to evaluate specificity and binding affinity. Functional interactions were assessed in Escherichia coli expressing PfHSP70 or PfLDH. Recombinant PfHSP70 and PfLDH were successfully expressed and purified from E. coli, with approximately 50% of selected colonies demonstrating significant binding to both targets, confirming the enrichment of antigen-specific phages. Specific phages were validated using ELISA and transmission electron microscopy (TEM). Selected scFvs exhibited strong target binding, with MST-determined dissociation constants (Kd) of 7.47 μM for PfHSP70 and 3.64 μM for PfLDH. Exposure to scFv-displaying phages impaired the survival of PfHSP70- and PfLDH-expressing E. coli and induced spectral shifts consistent with protein-antibody interactions. This study validates phage display as a robust platform for isolating high-affinity scFvs against P. falciparum targets. These binders have the potential to develop into low-cost diagnostic tools, addressing the urgent need for novel diagnostic interventions. Nevertheless, further studies are required to confirm binding specificity and assess translational applicability.
Pomelo (Citrus maxima) peel essential oil is a volatile-terpenoid-rich plant extract that may exhibit antioxidant-associated effects in biological systems. However, the relationship among its chemical composition, conventional antioxidant capacity, and cellular effects under oxidative stress remains insufficiently defined. This study integrated gas chromatography-mass spectrometry (GC-MS) profiling with chemical antioxidant assays and an SH-SY5Y cell-based oxidative-stress model. Under the selected analytical and data-processing conditions, limonene was the only compound confidently identified and integrated by GC-MS. In chemical assays, pomelo peel essential oil showed moderate DPPH radical-scavenging and ferric-reducing activities compared with the hydrophilic reference antioxidants Trolox and ascorbic acid. Cellular effects were evaluated separately in SH-SY5Y cells exposed to H2O2-induced oxidative stress. Pomelo peel essential oil was tested at 25, 50, and 100 μg/mL using cotreatment and pretreatment protocols. In the cotreatment model, cells were exposed simultaneously to the essential oil and H2O2, whereas in the pretreatment model, cells were incubated with the essential oil before H2O2 challenge. Pomelo peel essential oil attenuated intracellular ROS accumulation and lipid peroxidation and improved cell viability, with clearer effects at 50 and 100 μg/mL. Pretreatment generally produced stronger cytoprotective effects than cotreatment. These findings indicate that conventional chemical antioxidant assays alone may not fully predict cellular antioxidant-associated effects, particularly for lipophilic essential oils. Pomelo peel essential oil therefore showed cytoprotective activity in an SH-SY5Y oxidative-stress model, although the contributing constituents and underlying molecular mechanisms require further investigation.
Repetitive traumatic brain injury (RTBI) can cause long-term complications, including persistent neuroinflammation, which can extend beyond the central nervous system, impacting various peripheral organs as liver. This study aimed to explore the neuroprotective and hepatoprotective effects of arbutin treatment in a rat model of mild RTBI (mRTBI), focusing on nerve growth factor (NGF)/tropomyosin receptor kinase A (TrkA) signaling pathway along with the crosstalk between brain injury and hepatic dysfunction. Animals were randomly assigned into three groups: one served as a normal control (NC) group, while the other two groups were exposed to one blow for 5 days and either left for one week after the fifth blow (mRTBI) or received arbutin intraperitoneally (100 mg/kg/day for 7 days, mRTBI + ARB). Biochemical and histopathological changes were monitored in the brain cortex and the liver. This study revealed that arbutin treatment offered neuroprotection and preserved most of the neuronal structures. Arbutin demonstrated a significant increase in the cortical NGF and TrkA contents, along with a marked upregulation in cortical phosphoinositol-3 kinase (PI3K) and protein kinase B (AKt) mRNA levels compared to the mRTBI group. Furthermore, arbutin decreased cortical and serum inflammatory markers, reflecting its anti-inflammatory power. Peripherally, arbutin treatment resulted in a substantial decrease in hepatic inflammatory markers, Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) and caspase-3. These effects preserved hepatocellular histoarchitecture and reduced liver injury markers. Collectively, arbutin effectively modulated the NGF/TrkA signaling pathway, diminished inflammation, and alleviated the detrimental effects of mRTBI on both the brain and liver.
