Applied Biochemistry and Biotechnology最新文献

HCC predominantly develops in individuals with chronic hepatic conditions and liver cirrhosis, which is marked by high mortality. This study investigates the functional roles and molecular mechanisms by which FOXP2 modulates HCC progression through ferroptosis. After HCC and normal cells were cultured, the expression of FOXP2, RBM15B, and KDM4C was analyzed using western blot or RT-qPCR. After FOXP2 intervention, cellular metabolic activity was assessed via CCK-8 assay, while replicative capacity was quantified through EdU staining and colony formation assays; key regulators of ferroptosis were analyzed by western blot; iron content and oxidative stress levels were measured. The binding between FOXP2 and RBM15B was investigated through ChIP and dual-luciferase assays. Dual-luciferase reporter assay was used to verify the regulation of RBM15B on KDM4C via m6A modification. MeRIP was utilized to examine m6A enrichment on KDM4C mRNA. ChIP was employed to examine the enrichment of KDM4C and H3K9me3 on SLC7A11 promoters. Combined experiments investigated the role of the RBM15B/KDM4C axis in FOXP2-mediated ferroptosis and HCC cell proliferation. Xenograft models were developed in nude mice to validate the mechanism. FOXP2 expression was downregulated in HCC. FOXP2 overexpression significantly inhibited HCC cell proliferation and promoted ferroptosis. FOXP2 repressed RBM15B expression, suppressed the RBM15B-mediated m6A modification, inhibited KDM4C expression, upregulated H3K9me3 levels, and suppressed SLC7A11 expression, ultimately enhancing ferroptosis. Overexpression of RBM15B or KDM4C attenuated ferroptosis and reversed the suppression of HCC cell growth induced by FOXP2 overexpression. In conclusion, FOXP2 may promote ferroptosis and inhibit cell proliferation in HCC by decreasing SLC7A11 expression via the RBM15B/KDM4C axis in an m6A-dependent manner.

Alzheimer's disease (AD) is a progressive neurodegenerative condition characterized by memory deficits, oxidative stress, and neuroinflammation. Plumbagin (PB), a bioactive naphthoquinone derived from Nepenthes khasiana, possesses therapeutic potential but is limited by poor solubility and bioavailability. To overcome these challenges, PB was encapsulated into hyaluronic acid nanogels (PB-NGs) and systematically evaluated for anti-Alzheimer's efficacy. The study aimed to evaluate the neuroprotective and anti-inflammatory properties of PB-NGs by in vitro and in vivo analyses. PB-NGs were synthesized and characterized using FT-IR, NMR, and TEM, confirming successful encapsulation. In silico docking demonstrated strong binding affinities of PB to β-amyloid and tau proteins, suggesting potential inhibition of AD-related pathological processes. In vitro, PB-NGs significantly decreased ROS production, mitigated pro-inflammatory cytokines (IL-1β, TNF-α), and improved cell viability in LPS-stimulated SH-SY5Y cells relative to free PB. In vivo, administration of PB-NGs to AD-induced mice significantly improved behavioral outcomes, including enhanced spatial memory and exploratory activity in Y-maze, open field, and Morris's water maze tests. Biochemical assays further revealed reduced oxidative stress (decreased MDA, increased SOD, CAT, GST) and modulation of cholinergic dysfunction (reduced AChE, increased ChAT). Histopathological analysis supported neuronal protection with diminished amyloid burden. Collectively, these findings demonstrate that PB-NGs effectively enhance bioavailability and exert significant neuroprotective and anti-inflammatory effects, supporting their potential as a promising nanotherapeutic approach for mitigating cognitive deficits in AD.

The misuse of antibiotics has led to the rise of multidrug-resistant (MDR) pathogens, posing a significant threat to global health. The shortcoming of new antibiotics with novel mode of action augments this challenge. Nanoparticles, particularly synthesized through green synthesis methods, have emerged as promising agents to combat the growing issue of MDR. The current study focuses on the green synthesis of silver nanoparticles (AgNPs) using seed extract from the traditional medicinal herbaceous plant Phyllanthus maderaspatensis (PM). AgNPs were synthesized by mixing the PM seed extract (PMSE) with 3 mM silver nitrate at 80 °C for 15 min, followed by precipitation using acetone and drying at 70 °C. Characterization of the derived AgNPs with UV spectroscopy resulted absorption maximum at 430 nm. FTIR analysis revealed the capping of functional moieties such as alcohol, amine, aldehyde, alkene and halo to their surfaces. SEM and TEM analysis disclosed the spherical and quasi-spherical shaped nanoparticles, with smooth surface and notable lattice fringes appearance. The size of AgNPs ranges from ~ 5 nm to ~ 78 nm in diameter. The synthesised nanoparticles happen to be highly stable as deduced via mean zeta potential of -37.9 mV. XRD and energy dispersive X-ray spectrum of the synthesized AgNPs conforms the presence of silver in the synthesised nanoparticles. The antimicrobial potential of the synthesized AgNPs against different bacterial strains provides minimum inhibitory concentration values as low as 150 to 225 µg/mL. Additionally, the AgNPs also exhibited outstanding anti-biofilm capabilities. Crystal violet uptake assay and light microscopy studies indicates that membrane disruption contributes to their bactericidal effect. Altogether, the utilization of PMSE as a reducing agent holds promise as a cost-effective, scalable, and eco-friendly alternative to traditional AgNP synthesis methods. This PMSE derived AgNPs demonstrate strong potential for broad applications namely in agriculture for management of PM seeds and across medicine such as development of anti-bacterial coating or as an active ingredient in wound dressing.

L-asparaginase (ASNase) is one of the most clinically relevant biopharmaceuticals but proteolysis impairs the enzyme half-life. ASNase P40S/S206C was developed to overcome proteolysis and in addition pegylation can be used to improve half-life and thermostability. Here Cys206 and N-terminal residues were explored as pegylation sites for a potential new biobetter. Optimal mono-pegylation of Cys (Cys-PEG-ASNase) provided similar yields compared to N-terminal mono-pegylation (NT-PEG-ASNase) but at lower PEG concentration. Specific activity was higher for Cys-PEG-ASNase than NT-PEG-ASNase. The role of pegylation site in activity was confirmed by activation energy (Ea) and enthalpy variation (ΔH_70ºC) of the ASNase-catalysed reaction. Pegylation in both sites increased enzyme thermostability and consequently shelf-life stability. The variation of Gibbs free-energy of enzyme thermo-inactivation (ΔG_d) showed higher stabilization by Cys conjugation, while enthalpy (ΔH_d) and entropy (ΔS_d) evidenced an increase in aggregation upon thermo-inactivation, higher for Cys-PEG-ASNase.

The advancement of methodologies for extracting biomolecules from natural sources has gained increasing attention in recent years because of their diverse applications, particularly in food and medicine. In the present study, a commercial pectinase enzyme was employed for the hydrolysis of lignocellulosic biomass, and the proposed mechanical extraction technique, aided by the enzyme, was proven to be advantageous for the extraction of pumpkin seed oil. An analysis of the physicochemical properties of pretreated pumpkin seed oil under different temperature conditions revealed that the density, refractive index, and iodine values decreased with increasing temperature, whereas the color indices, peroxide values, and free fatty acids were directly correlated with temperature. Proximate analysis revealed that the protein content was highest in seeds treated with microwave-assisted enzymes (25.77%), with a crude fibre content of 5.05 g per 100 g of seeds. The ash content of untreated pumpkin seeds was measured at approximately 3.6 mg per 100 g, with the enzyme indirectly impacting the ash levels in the seeds. The addition of pectinase to the mechanical extraction process increased the antioxidant activity of pumpkin oil, as evidenced by a total phenolic content (TPC) of 19.08 mg GAE per gram, and increased the yield from 28.25% to 30.85%.

The emergence of insecticide resistance in pests affecting cotton and other key crops presents an escalating challenge to global agriculture, resulting in considerable declines in both yield and quality. Resistance mechanisms, including target site mutations and improved detoxification processes, have diminished the effectiveness of traditional insecticides. This study identified NPA001937 (di-O-demethylspirilloxanthin), a bacterial carotenoid obtained from the Natural Products Atlas, as a promising natural insecticidal compound. Employing in silico methods, we focused on three essential insect targets: Ryanodine Receptor (RyR), Arginine Kinase (ArgK), and Serine/Threonine Protein Phosphatase (STPP), which are known for their conserved sequences and structures across various pest species. Structural models were produced using AlphaFold2 and SwissModel, followed by validation through SAVES. The virtual screening of 30,052 natural products conducted with AutoDock Vina revealed NPA001937 as a leading binder to all three targets. Subsequent 500 ns molecular dynamics simulations (GROMACS) validated the stability and advantageous interaction profiles of the ligand- protein complexes. This compound presents several advantages over chemical insecticides, including its natural origin, ability to target multiple pathways, and a lower chance of resistance development. The results emphasize the effectiveness of computational methods in speeding up the discovery of agrochemicals which upon further experimental validation may serve as ecofriendly options for sustainable pest management.