RSC Advances

Natural products are a rich source of bioactive molecules and undiscovered chemical scaffolds with significant potential for novel drug discovery. Among these, fungi are particularly promising, offering diverse metabolites and undiscovered structural motifs. Large, well-curated collections of crude extracts, or "libraries", are central to fungal natural product discovery, serving as starting material for bioassay-guided isolation of new compounds. However, the systematic influence of fungal selection strategies, culturing methods, and environmental factors on chemical diversity remains underexplored. In this study, we analyzed several large fungal libraries to assess how geographic origin, and phylogenetic classification shape fungal chemical profiles. We also evaluated whether culturing conditions that more closely mimic natural environments can enhance metabolite diversity. Our findings offer practical guidelines for optimizing fungal natural product library design, improving drug development efficiency and access to novel chemotypes for future drug discovery. Summary Figure O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=60 SRC="FIGDIR/small/709592v1_ufig1.gif" ALT="Figure 1"> View larger version (16K): org.highwire.dtl.DTLVardef@70a0e0org.highwire.dtl.DTLVardef@51f84eorg.highwire.dtl.DTLVardef@184dd90org.highwire.dtl.DTLVardef@1ee2813_HPS_FORMAT_FIGEXP M_FIG C_FIG

Microbial halogenated natural products (hNPs) hold ecological, agricultural, and biomedical relevance. The hNP-producing potential of the organism can be assessed by the precise prediction of biosynthetic enzymes, yet the detailed annotations of halogenases are often missing from genomic and metagenomic data. We created a manually curated database (https://halogenases.secondarymetabolites.org/) containing information on the halide-specificity, role, and position of verified catalytic residues and results of the mutagenesis studies of more than 120 experimentally validated or in silico inferred halogenases. The collection of experimental data supports a computational pipeline that allows the family-, substrate-, and halide-scope-level annotation of halogenating enzymes by relying on catalytic residues, conserved motifs, and profile Hidden Markov Models (pHMMs). Our analysis with sequence similarity networks (SSNs) highlighted several underexplored clusters in the UniRef50 database. Such finding was a halogenase from Rhodopirellula baltica (RhobaVHPO) previously labelled as a hypothetical chloroperoxidase, which clustered apart from the known chloroperoxidases and bromoperoxidases, but accepted chloride and preferred bromide. Our database and workflow provide extensive and scalable solutions for the systematic and precise annotation of halogenating enzymes in genomic and metagenomic data. The in-depth categorization of halogenases will improve the chemical structure prediction of microbial hNPs, supporting ecological assessments and natural product discovery. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=112 SRC="FIGDIR/small/700248v1_ufig1.gif" ALT="Figure 1"> View larger version (45K): org.highwire.dtl.DTLVardef@ebae51org.highwire.dtl.DTLVardef@10188f0org.highwire.dtl.DTLVardef@1c55684org.highwire.dtl.DTLVardef@b311bd_HPS_FORMAT_FIGEXP M_FIG C_FIG

Lung cancer remains the leading cause of cancer-related deaths worldwide, with cisplatin as the primary chemotherapy despite its limitations. Organotin(IV) dithiocarbamates have emerged as promising anticancer agents due to their potent cytotoxicity and stability. This study reports the successful synthesis of four novel organotin(IV) dithiocarbamates: dimethyltin(IV) N-methyl-N-benzyldithiocarbamate (DioSn-1), diphenyltin(IV) N-methyl-N-benzyldithiocarbamate (DioSn-2), triphenyltin(IV) N-methyl-N-benzyldithiocarbamate (TriSn-3), and triphenyltin(IV) N-ethyl-N-benzyldithiocarbamate (TriSn-4). Their cytotoxicity against A549 lung carcinoma cells was evaluated via MTT assay, while Annexin V-FITC/PI staining determined the mode of cell death. DioSn-2, TriSn-3, and TriSn-4 exhibited potent cytotoxicity (IC: 0.52-1.86 M), whereas DioSn-1 was inactive (IC > 50 M). Apoptotic features such as cell shrinkage and membrane blebbing were observed, with apoptosis rates ranging from 58% to 91%. DioSn-2 was the most selective (SI = 6.45) and induced early DNA damage within 30 minutes, followed by mitochondrial depolarization and excessive ROS generation. Caspase-9 activation exceeded caspase-8, confirming intrinsic apoptosis. NAC treatment reduced apoptosis by 52%, highlighting oxidative stress as a key cytotoxic mechanism. These findings suggest DioSn-2 as a promising alternative to cisplatin for lung cancer therapy.

In siRNA-based applications, cellular delivery remains one of the main hurdles. Many formulations are tested for the same and peptides came up as one of the optimal options. The latter have various advantages like natural biological presence, high specificity, and natural metabolism etc. siRNA in conjugation with peptides have exhibited enhanced mRNA silencing. Peptides aid siRNAs in condensation to smaller volumes, enhance nuclease protection, increase half-life, promote cell specific binding as well as endosomal escape and release in cytosol. Despite its prime importance, no resource is available for the peptide-based delivery of siRNAs, therefore to fill the gap we developed PEPsRNA web server. It includes 2266 entries of 270 different kinds of peptides, 106 different types of siRNAs and shRNAs along with more than 80 conjugate molecules targeting 55 different genes, experimentally tested for the delivery of the siRNAs. To provide the detailed insights of the procedure, we have incorporated analysis of the peptides (e.g. secondary structure, amino acid composition, polarity, hydrophobicity etc.), siRNAs (e.g. secondary structures with minimum free energies etc.) and associated conjugate molecules (e.g. structure, SMILES, Inchl). We have derived these values using various other tools and resources to make the web server comprehensive. We further compared various physicochemical properties with the efficacy of the peptide based on the target gene silencing, but these properties do not shown any distinct conclusive relationship. The data is available for browsing, searching and downloading freely on the web server with URL: http://bioinfo.imtech.res.in/manojk/pepsirna. Highlights PEPsRNA is the first database of experimentally tested peptides for siRNA delivery It comprised of 2266 entries with 270 peptides and about 80 conjugate molecules Analysis of peptides, siRNAs and details of conjugate molecules are provided Browse, search and various tools are incorporated for data retrieval and usage

Globally, about 264 million individuals across all age groups are impacted by depression, a prevalent central nervous system (CNS) condition. Chronic and enduring depression might result in significant health consequences. Numerous pharmaceutical antidepressants exist for the management of mild to severe depression, largely functioning by modifying neurotransmitter levels in the brain. Nevertheless, these drugs frequently induce a variety of side effects, such as insomnia, constipation, exhaustion, drowsiness, and anxiety. Saffron (Crocus sativus L.) is widely acknowledged as a natural antidepressant with little adverse effects. This study investigated the potential antidepressant mechanisms of saffrons principal bioactive compounds safranal, crocin, and picrocrocin via molecular docking against critical target proteins associated with depression, namely the dopamine transporter (DAT), serotonin transporter (SERT), and monoamine oxidase B (MAO-B). Molecular docking was conducted with AutoDock 4.2 to assess the binding affinity and interaction energy of these drugs with the target proteins. Furthermore, Discovery Studio facilitated the viewing and study of both interacting and non-interacting residues at the docking sites, juxtaposing these interactions with those of established inhibitors in crystal structures. The permeability of the blood-brain barrier (BBB), pharmacokinetic characteristics, and toxicity profiles of saffron components were evaluated using SWISS ADME, DataWarrior, and Osiris Molecular Property Explorer. Among the evaluated elements, safranal had the greatest potential as a competitive inhibitor of the dopamine transporter, according to its notable blood-brain barrier permeability, robust binding affinity, and analogous interaction residues in comparison to nortriptyline, a recognized inhibitor. Our findings indicate that safranal may be a viable natural alternative to traditional antidepressants, with minimized adverse effects.

Accounting for 12% of global solid waste, Poly (ethylene terephthalate) (PET) is one of the most abundantly produced synthetic polymers. While PET offers substantial commercial benefits, its widespread use has led to disproportionate environmental hazards due to its resistance to degradation. To address this problem, several solutions have been proposed, including enzymatic degradation via PETase, MHETase, and Cutinase. Among these, PETase exhibited significant PET-degrading activity. However, the application of PETase has been hampered by its lack of robustness to pH, temperature ranges, and slow reaction rates. Hence, it has become novel enzymes that can overcome these limitations and function efficiently. In this study, we utilised an integrated in silico bioinformatics pipeline to identify and characterise novel PETase candidates from the Thermophilic actinobacteria Thermobifida cellulosilytica and Thermobifida halotolerans species. The PlasticDB database contains 228 plastic-degrading enzyme sequences. In which PETase (00188) is significantly homologous with two putative proteins, Hydrolase (ALF00495.1) and hypothetical protein (WOZ56011.1). The discrete optimized protein energy (DOPE) scores, stereochemical assessments, and homology modeling results closely mirrored our findings for both proteins, supporting their structural stability. The molecular dynamics simulations revealed that the putative ALF00495 variant exhibited more extensive and robust hydrogen-bonding networks, enhanced conformational stability, and increased structural compactness compared to the reference enzyme. The present in silico investigation underscores the potential of putative ALF00495 as a highly effective PETase biocatalyst for polyethylene terephthalate (PET) degradation. Collectively, these findings illustrate the utility of computational approaches of novel PET-degrading enzymes, thereby facilitating the development of sustainable biotechnological strategies to mitigate global plastic pollution.

Metallic nanoparticles have emerged as novel therapeutic agents due to their distinctive physicochemical properties and broad-spectrum activity, with applications in antimicrobial therapy, drug delivery and bioremediation. Conventional methods for metallic nanoparticle synthesis often utilize toxic chemicals and energy intensive processes that are expensive. Green synthesis offers a sustainable and cost-effective alternative by using biomolecules from plants and microorganisms. In this study, gold (AuNPs), silver (AgNPs), and copper oxide (CuO NPs) nanoparticles were biosynthesized using leaf extracts of Aloe africana Mill., a South African medicinal plant rich in phytochemicals, and the magnetotactic bacterium Magnetospirillum magnetotacticum that naturally produces intracellular nanoparticles. GC-MS analysis revealed 13 known phytochemicals in the A. africana extract including esters, terpenoids, monoglycerides, and fatty acids which served as reducing and capping agents for nanoparticle synthesis. A. africana-derived AgNPs were spherical (11-30 nm) in shape, capped with dihydrosqualene, a known antibacterial compound; and was found to display activity against Gram-negative (Escherichia coli and Pseudomonas aeruginosa) and Gram-positive (Enterococcus faecalis and Staphylococcus aureus) bacteria. These AgNPs however exhibited cytotoxicity to HEK293 and HeLa cell lines. A. africana AuNPs (17-62 nm) displayed diverse morphologies and CuO NPs (55-115 nm) were irregular shaped, and both nanoparticles exhibited limited antibacterial activity and low cytotoxicity. M. magnetotacticum-derived AuNPs (12-21 nm) and AgNPs (51-126 nm) were spherical, with the CuO NPs (42-66 nm) having irregular shapes. Except for A. africana-derived AgNPs, all other metallic nanoparticles displayed poor antibacterial activity. These findings are novel and highlight a dual-function green synthesis platform where A. africana phytochemicals contribute to both nanoparticle synthesis and bioactivity, positioning A. africana AgNPs as promising antibacterial agents.

Antimicrobial resistance (AMR) in plant pathogenic bacteria poses a serious threat to global agriculture, necessitating the development of novel antibacterial agents targeting virulence mechanisms. This study presents an integrated bioinformatics-driven framework for the rational design and computational validation of Solres, a newly designed small molecule targeting key virulence proteins in phytopathogenic bacteria. Approximately 10,000 active compounds from PubChem BioAssay (AID: 588726) were analyzed using structural clustering and scaffold mining to identify conserved molecular motifs associated with antibacterial activity. Guided by high-frequency substructures, Solres was designed de novo and screened for structural novelty against PubChem, ChEMBL, and WIPO databases. Drug-likeness evaluation using Lipinskis Rule of Five confirmed favorable physicochemical properties. Molecular docking was performed against essential virulence regulators, including PhcA, PhcR, HrpB, PehA, and Egl from Ralstonia solanacearum and Xanthomonas spp., with active sites predicted using CaspFold. Docking analyses revealed strong binding affinities and stable interactions with key catalytic and regulatory residues. Complex stability and conformational integrity were further validated through molecular dynamics simulations. Quantum chemical descriptors, including HOMO-LUMO energy gap and dipole moment, supported the electronic suitability and reactivity profile of Solres. Collectively, this study demonstrates the effective integration of cheminformatics, structural bioinformatics, molecular simulations, and quantum chemical analyses for plant-focused antibacterial discovery. The compound Solres represents a promising lead candidate for mitigating bacterial wilt disease and provides a computational framework for future experimental validation and sustainable crop protection strategies against AMR-driven phytopathogens.

Infectious diseases remain a persistent global health burden, with bacterial infections predominating. The growing global burden of drug-resistant infections has led to greater emphasis on the discovery and development of novel antibacterial compounds. In an attempt to discover new potent antibacterials, the antibacterial activity of novel 2-substituted benzimidazole derivatives (NR-1 to NR-9) was evaluated in this study against three bacteria, viz. M. smegmatis, B. subtilis and E. coli in vitro using Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC). Among the nine derivatives assessed, two (NR-4 and NR-5) exhibited inhibitory activity against M. smegmatis, while two (NR-5 and NR-7) were active against B. subtilis, with MICs between 62.5 and 250 g/ml. Notably, NR-5 demonstrated antibacterial activity against both M. smegmatis and B. subtilis, with more efficacy against M. smegmatis (MIC: 62.5 g/ml), which was considerably closer to rifampicin (MIC: 31.25 g/ml). Cytotoxicity analysis of these derivatives in Vero cells indicated minimal toxicity for NR-4 and NR-5, and SwissADME evaluation suggested favourable physicochemical properties and drug-likeness, supporting good oral bioavailability. Moreover, the growth kinetics profiling of the NR-5 Benzimidazole derivative demonstrated that it inhibited the growth of M. smegmatis effectively, even after prolonged exposure. These findings highlighted the promise of the active benzimidazole derivative, NR-5, as a potential candidate for developing a more effective and less toxic antimycobacterial drug. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=76 SRC="FIGDIR/small/710429v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@126a0fcorg.highwire.dtl.DTLVardef@1131372org.highwire.dtl.DTLVardef@161a70corg.highwire.dtl.DTLVardef@1e0dbe_HPS_FORMAT_FIGEXP M_FIG C_FIG

We have conducted all atom molecular dynamics simulations of POPC and DPPC lipid bilayers using AMBER Lipid21 force field with eight different water models, including SPC/E, TIP3P, TIP3P-FB, TIP4P-FB, TIP4P-Ew, TIP4P/2005, TIP4P-D, and OPC, to identify the most compatible one without any modification. A number of parameters have been computed in order to understand the structure of the lipid bilayer: Area per lipid, Isothermal compressibility modulus, average Volume per lipid, electron density profile, bilayer thickness, X-ray and neutron scattering form factors, deuterium order parameter, and radial distribution function. The estimated Area per lipid, Isothermal compressibility factor, volume per lipid and bilayer thickness are highly consistent with experimental results for the SPC/E water model, indicating its suitability with the AMBER Lipid21 force field, insted of any modification. The bilayer electron density profiles of both the lipid bilayers demonstrate a little augmentation of water penetration with respect to the membrane surface for TIP4P-D water model. However, the experimental X-ray and neutron scattering form factors are aligning well with the simulated results for all studied water models, and TIP4P-D shows better for X-ray data. The deuterium order parameter for lipid acyl chains value less than 0.25 for all observed water models, depicting their disorderness for both the lipid bilayers. The lateral diffusion and reorientation autocorrelation function of the lipid molecules in both the bilayers are computed to reveal their dynamics across all water models. In comparison to other water models, the simulated trajectories predict better structure and reasonably fair dynamic properties for the SPC/E water model. The TIP4P-Ew water model reproduces the lateral diffusion co-efficient in close agreement with experiment. Reorientational dynamics for both the lipids in the bilayers for eight different water models are observed; the presence of slow and slowest time components corresponds to the lipid axial motion (wobble motion) and Twist/Splay motions. So, in view of the overall performance of the different water models with the AMBER Lipid21 all atom force field in reproducing membrane physical properties, the SPC/E water model appears to be an optimal choice.

Forest tree seeds are mass produced for afforestation and forest restoration programs, but are mostly underutilized beyond propagation. Here, we aimed to evaluate the antioxidant, anti-inflammatory, anticancer, and tyrosinase-inhibitory activities of seed extracts of seven economically important forest tree species in the Republic of Korea to explore their potential as multifunctional natural bioresources. The seed extracts of Alnus japonica, Chamaecyparis obtusa, Cornus kousa, Phellodendron amurense, Pinus densiflora, Prunus sargentii, and Quercus glauca were comparatively assessed using multiple in vitro assays. The results revealed clear species-dependent functional profiles rather than uniform bioactivities across species. Quercus glauca exhibited strong antioxidant activity and significant anti-inflammatory and tyrosinase-inhibitory activities, suggesting multifunctional potential, while C. obtuse presented considerable anticancer activity against several cancer cell lines. Alnus japonica exhibited the highest tyrosinase-inhibitory activity, followed by Q. glauca and C. obtuse; A. japonica extract also showed a strong antioxidant capacity. Overall, the results demonstrated that forest tree seed extracts possess diverse and complementary bioactivities, supporting their potential as underexplored multifunctional natural materials. By focusing on seed resources generated within existing afforestation systems, we highlight a sustainable approach to valorize forest-derived by-products without additional pressure on natural ecosystems. Nevertheless, as bioactivities were evaluated using crude extracts, further studies are required to identify and elucidate the active compounds and their mechanisms of action.

Polyunsaturated fatty acids (PUFAs) are fundamental for different cellular and structural processes, especially regarding the nervous system. However, its incorporation in food has many bioaccessibility limitations, making it important to find new ways of its consumption. In this work, nanoencapsulated PUFAs orally administrated to C57BL/6 elite male mice for 8 weeks showed better bioavailability when compared to administration of free acids, also improving the effects on dentate gyrus neuronal survival as well as on the proneurogenic elements of the Brain derived neurotrophic factor biological pathway also in the hippocampus. In addition, nanoencapsulated PUFAs increased expression of Fabp5, a relevant n-3 fatty acids transporter in the brain. Altogether, our results would mean that the form of administration of the fatty acids can alter not only how much and how preserved they reach the central nervous system, but also have a differential impact in the diverse processes they contribute to.

Current in silico drug discovery protocols ubiquitously depend on lead generation using a ligand-based approach in which novel leads are generated by fragment-signature matching or by a structure-based search involving molecular docking and conformational dynamics. None of them incorporates cellular contexts in which these drugs ultimately operate, leaving the task to a later stage of optimization leading to a high failure rate. Incorporating systems-level responses of drugs in an early stage of lead generation can significantly address this concern but has not been sufficiently explored. In this work, we employ a systems-level approach using connectivity map (CMAP) library to generate leads against a challenging system of a TLR pathway. Starting with gene expression data of TLR5 activation by its natural ligand, we generated molecular leads using CMAP and rigorously analyzed their validity using ligand and structure-based approaches, and helping to prioritize top hits. Experimental validation using ELISA-based antibody assay confirmed the activation of TLR5 by each of the top nine prioritized leads with their dose-dependent patterns suggesting that some of them may actually interact with the TLR signaling pathway in a complex manner. Although, demonstrated on TLR5, the proposed framework is intuitively scalable to other lead generation and optimization tasks.

Globally, Mycobacterium tuberculosis remains a significant disease burden. Although effective treatment regimens exist, drug resistance continues to emerge. This clinical resistance, combined with side effects and protracted treatment times from the current front-line therapies, means there is a need to identify novel agents to combat this disease. Here we report on a new chemical series, identified by whole-cell phenotypic growth inhibition screening that demonstrates significant activity across multiple media. Mode of action studies indicate that this series targets the same biological pathway as Ethambutol (EMB), a drug used in the current frontline treatment of tuberculosis. Screening selected analogues against clinical isolates, resistant to EMB, demonstrated differential sensitivity both across the molecules and against the different specific resistant mutations. The data obtained suggests that this series has potential to be developed into a viable, alternative to EMB. TOC figure O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=93 SRC="FIGDIR/small/702510v1_ufig1.gif" ALT="Figure 1"> View larger version (14K): org.highwire.dtl.DTLVardef@1a80c05org.highwire.dtl.DTLVardef@1ad3ce9org.highwire.dtl.DTLVardef@79fe79org.highwire.dtl.DTLVardef@131ed78_HPS_FORMAT_FIGEXP M_FIG C_FIG

The production of petroleum-based plastics used for packaging has led to significant environmental challenges in both aquatic and terrestrial ecosystems. Consequently, there is a growing need to explore viable alternatives to the usage of these conventional plastics. This study investigates the utilization of cellulose powder for producing of biodegradable plastics as a more sustainable substitute for petroleum-based materials. Bioplastic films were formulated with varying glycerol contents ranging from 0.5ml - 2.0ml. The glycerol served as a plasticizer to improve the mechanical properties of the films, which were subsequently subjected to biodegradability and tensile strength tests. Biodegradability was evaluated through soil burial tests, which revealed that higher glycerol concentrations accelerated rate of weight loss, with the 2.0 ml formulation exhibiting the fastest degradation rate. Tensile strength increased with glycerol content up to 1.5 ml, where a maximum strength of 7.23 N/mm2 was recorded, but declined at 2.0 ml. The findings indicate that a glycerol concentration of 1.5 ml yields the most optimal bioplastic formulation for short-term packaging applications.

Promyelocytic leukemia (PML) proteins are known to form phase-separated nuclear punctate structures called PML-nuclear bodies (PML-NBs). The integrity disruption of PML-NBs is linked with the pathogenesis of acute promyelocytic leukemia (APL), and trivalent arsenic (As3+) has been used for the clinical treatment of APL to restore normal PML-NBs. As3+ is considered to bind to cysteine residues and enhances modification of PML with small-ubiquitin-like protein (SUMO). We exposed U-2OS and CHO-K1 cells stably overexpressing PML-VI to As3+ and found that the solubility of PML decreased and SUMOylation of PML increased after 2 h-exposure to 3 M As3+. Contrary to As3+-induced remarkable biochemical changes including the solubility change and SUMOylation of PML, microscopic observation of PML-NBs was not changed clearly after a short-term exposure to As3+. The number of PML-NBs decreased and extranuclear PML bodies (EnPBs), which are remniscences of PML-NBs after nuclear membrane breakdown at mitosis, increased after exposure to As3+ for 24 - 72 h. The amount of SUMOylated PML decreased after prolonged exposure to As3+ while the solubility of PML was kept low, suggesting that As3+ stabilized EnPB without SUMOylation. The effects of As3+ on EnPBs were clearly observed at as low as 0.3 M As3+ which corresponds to inorganic arsenic level in drinking water worldwide.

Molecular recognition is a central component that confers detection specificity to all biosensors. The design and use of such molecules require consideration of properties including their affinity and selectivity, plus their ease of production and engineering, for downstream commercial purposes. Progesterone (P4), is a biomarker that is extensively for various diagnostic purposes. Examples include detection of P4 as an indicator of oestrus in cattle breeding, and ovulation in human IVF programs. P4 is also thought to promote strains of breast cancer, resulting in it being an environmental pollutant of interest. The present study focusses on in-silico molecular docking trials of P4 molecules with proteins such as antibodies and receptors. We describe the geometry of novel P4-binding pockets and predict key residues that favour high affinity and selectivity for P4. The in-silico molecular docking trials were performed on various mutants of an anti-P4 antibody that had lost their P4 specificity but retained selective recognition of steroids with structures closely related to cholesterol. Reverse-docking trials permitted the identification of novel scaffolds with favourable P4 binding properties. Future reports will validate the predictions of these studies through wet lab experiments. A further opportunity for this approach is to incorporate a scaffold functionality to permit binding of the protein or receptor to other molecules or sites within a biosensor electrode. These findings, and future studies, will assist in development of enhanced biosensing platforms with custom-designed P4 binders, aiding commercialisation using in-house developed reagents to meet IP requirements and minimise scaling costs. The steroid biotechnology market, valued at over $10 billion, also benefits from novel steroid binder designs, facilitating real-time steroid biomonitoring platforms for optimising steroid bioprocesses.

Head and neck squamous cell carcinoma (HNSCC) is a therapeutically challenging cancer what underscoring the need for new chemical agents that selectively induce programmed cell death. Fisetin, a naturally occurring flavonoid, exhibits promising anticancer activity but displays limited proapoptotic efficacy and selectivity. Here, we examined whether alkoxylated modification of fisetin enhances its ability to induce apoptosis in HNSCC cells. Fisetin derivatives bearing four-carbon substituents were synthesized and evaluated in multiple HNSCC cell lines. Two derivatives, MKT218 and MKT257, markedly reduced HNSCC cell viability at low micromolar concentrations with low toxicity towards normal human fibroblasts. Notably, the observed cytotoxicity was not associated with activation of a canonical DNA damage response, as neither {gamma}H2AX accumulation nor p53 activation was detected. Furthermore, PARP1 cleavage and live-cell imaging combined with annexin V/EthD-III staining revealed a significantly higher proportion of apoptotic cells. The effect was stronger following treatment with MKT218 and MKT257 compared with fisetin. Time-lapse microscopy further demonstrated that fisetin derivatives, particularly MKT218, promote mitosis-associated apoptosis, in contrast to the predominantly cytostatic effect of fisetin. Moreover, in silico docking suggested that MKT218 exerts its pro-apoptotic activity through a multi-target interaction profile involving key regulators of cell survival and apoptosis rather than a single dominant target. To sum up, our findings suggest that alkoxylated fisetin derivatives may be constituted as new non-genotoxic inducers of apoptosis in HNSCC cells.

Three-dimensional (3D) bioprinted liver scaffolds offer a promising platform for drug screening, disease modelling, and regenerative medicine, yet their broader adoption is limited by the absence of robust post-fabrication preservation strategies. This study aimed to evaluate the impact of -80{degrees}C (deep freezer) preservation and evaluate the structural integrity and hepatic functionality of GelMA-decellularized liver extra cellular matrix (dECM)-based 3D bioprinted liver scaffolds. Bioinks were formulated using synthesized GelMA and solubilized rat liver dECM, and 3D scaffolds were fabricated via extrusion bioprinting into rectilinear grid scaffolds. The 3D scaffold preservations was performed by immersion into two different medium (the culture DMEM media and the other FBS-DMSO cocktail) was evaluated using MTT viability assay, and albumin assay. Preserved 3D bioprinted scaffolds retained overall architecture and cell distribution in the FBS-DMSO cocktail demonstrated by the live dead assay. Together, the data demonstrate that -80{degrees}C storage can maintain the basic cell viability ([~]80%) and a substantial fraction of liver-specific functionality in 3D bioprinted scaffolds but also highlight sensitivity to preservation-induced injury. These findings underscore the need for further optimization of cryoprotectant formulations and freezing protocols tailored to 3D bioprinted liver scaffolds, and provide a foundational framework for developing ready-to-use, cryopreserved 3D liver models for translational applications.

Industrial waste containing hydrophobic pollutants, like oils and hydrocarbons, is toxic and difficult to degrade, posing both ecological and human health risks. Biosurfactants are eco-friendly surface-active compounds produced by microorganisms, known for their ability to lower surface and interfacial tension, enhancing the solubility and bioavailability of hydrophobic compounds, facilitating their breakdown. The current study focuses on isolating biosurfactant-producing bacteria from industrial waste sources near Dhaka, Bangladesh, and characterizing their properties, determining potential usage. Using diesel-enriched nutrient agar, bacterial strains were isolated and screened for biosurfactant production by oil displacement, emulsification index (E24%), and drop collapse assay. The most promising isolates were characterized according to their biochemical activities and 16S rRNA amplicon-based sequencing. Isolation and characterization of the surfactants have been carried out using chromatographic techniques. The identified bacteria passed the drop collapse and oil displacement tests. CTAB agar assay, indicates their anionic nature, showing an emulsification index ranging 10-41%. The potential biosurfactant producers belong to Bacillus, Pseudomonas, Acinetobacter, and Enterobacterium genera. The surfactants showed antibacterial, antifungal, and plant growth promotion activity and have been characterized in terms of pH stability, salinity, adhesion, and temperature tolerance. The study successfully identified and characterized potential biosurfactant-producing bacteria from industrial waste, highlighting their efficiency in breaking down hydrophobic pollutants and hydrocarbons. These microorganisms provide a green and economical substitute for synthetic surfactants due to their biodegradability and lower toxicity. Upon further research and scaling, these bacteria can be a good source of biosurfactants for potential applications in industrial, agricultural, and biomedical fields. IMPORTANCEThe study carries high significance as it creates multi-disciplinary scopes for utilizing these environmentally adapted biosurfactant-producing bacteria in industry, agriculture, and medicine. Since the bacterial isolates have hydrocarbon degradation ability, upon optimization for higher production, industrial usage in oil refinery and other industries can be adopted. Due to their biodegradable nature, usage in wound healing bandages and as antimicrobial agents in medicine will be noteworthy. The isolates have plant growth promotion ability and utilizing them as biofertilizer will reduce the dependency on chemical fertilizers. This is the first detailed report on biosurfactant-producing bacteria from this industrial waste-polluted Turag River of Dhaka City. Moreover, it compiles detailed screening protocols and methods for analyzing such environmentally friendly microbes. Such characterization also opens the scope for optimizing the production of the surfactant compounds on a large scale and utilizing them commercially.