Natural compounds are crucial in pharmacological research for modulating numerous biological processes. This is particularly helpful in the case of complex diseases and in overcoming challenges (e.g., drug resistance and adverse effects). Berberine, an isoquinoline alkaloid extracted from plants such as Berberis, Coptis, and Hydrastis species, exhibits a wide range of activities, including antidiabetic, anticancer, anti-inflammatory, antimicrobial, and antiviral. However, due to its low oral bioavailability (poor absorption, high first-pass metabolism, along with rapid elimination), further mechanistic understanding and optimization are required. This review brings together molecular docking data to identify berberine's major targets implicated in various diseases, evaluate its multifunctional therapeutic potential, while exploring the benefits, drawbacks, as well as future research avenues of the methodology. A systematic search of databases, including PubMed, Scopus, Web of Science, ScienceDirect, and Google Scholar (as of March 2026), was conducted to identify docking studies providing binding affinity and interaction details. Target scores, Protein Data Bank (PDB) IDs, software/scoring functions (e.g., Auto Dock/Vina, Glide, GOLD, Discovery Studio), and validation methods (e.g., redocking, Root-Mean-Square Deviation (RMSD), and important residues that interact with the target were extracted. Berberine showed good predicted affinities at metabolic targets (between -5.4 and -10.5 kcal/mol) with the highest affinities for AKT1 (3O96; -10.5 kcal/mol) and GLUT4 (-10.1 kcal/mol), which included stabilizing contacts such as GLN79, LYS268, and VAL270 in AKT1 and inflammatory mediators such as TNF-α, TYR103, and TRP107 in GLUT4. Antimicrobial docking studies showed good binding to DNA polymerase III (2POL, -5.32 kcal/mol; GLY219) and FtsZ (8GZY, -6.59; FTSZ), notably FTSZ (3VDB, -38.95 kcal/mol; multiple hydrophobic residues). Anticancer targets included BRAF (1UWJ; -9.53) and exceptionally low energies for Aldose Reductase (ADR) (-48.17 kcal/mol; MM-GBSA), phosphodiesterase 4D (-41.58 kcal/mol), Glucocorticoid Receptor (GR) (-37.08), DHODH (-34.81 kcal/mol), and PDK1 (-31.31 kcal/mol), implicating modulation of the MAPK and PI3K/AKT pathways. Antiinflammatory docking found the top binders to be COX-2 (4COX; -10.16 kcal/mol) and PTGS2 (to -9.0 kcal/mol), while antiviral studies identified SARS-CoV-2 Mpro (6LU7; -9.65 kcal/mol; residues including HIS41, CYS145, and GLU166) and other proteins binding to PLpro, RNAdependent RNA polymerase (RdRp), Spike, alongside Dengue proteins. Multitarget complex formation is promoted by recurring hydrogen bonding, hydrophobic, electrostatic, and π-π interactions, all of which are consistent with the planar, positively charged tetracyclic scaffold of berberine (C20H18NO4 +, 336.38 g/mol). There is methodological variation in docking protocols across studies, and the concordance between scores and activities remains inconsistent; however, datasets with identical parameters remain comparable. Overall, docking evidence reveals berberine to be a multitarget, broad-spectrum lead molecule and further supports the necessity of validating the molecule through molecular dynamics studies, experimental trials, clinical trials, and by designing derivatives, using nanotechnology for delivery, with artificial intelligence optimization, and in combination with other drugs to overcome its pharmacokinetic limitations and transform computational results into therapeutic potential.

Enzyme immobilization is frequently used in catalytic processes as an alternative to improve enzyme activity and stability. Among the materials used as a support, nanomaterials have been successfully reported in the literature, mainly due to its high specific area. In this study, the nanomaterials graphene, reduced graphene oxide, boron nitride nanosheets, and gold nanoparticles were evaluated as a support for pepsin immobilization. The effectiveness of this process was evaluated by the content of immobilized protein and activity of immobilized pepsin, determined through casein hydrolysis, as well as circular dichroism to verify possible alterations in enzyme structure. The antioxidant properties of hydrolysates were assessed by DPPH, ABTS, FRAP, and β-carotene. All the materials studied presented a high efficiency of immobilization, with over 80% of the pepsin content bound to the support, the use of graphene derivatives as support led to a reduction in proteolytic activity, with the immobilized pepsin retaining only 6% of its activity. The 45 nm gold nanoparticle led to an increase in proteolytic activity from 0.050 to 0.104 U. No effect of amino group functionalization of boron nitride nanosheet was observed. The variations determined in the secondary structure, especially in the β-sheet content, corroborate the variations in activity after immobilization. The change in casein hydrolysates using pepsin immobilized in nanomaterials can be verified by reversed-phase chromatography. This change in the enzyme conformation affects the antioxidant properties of the casein hydrolysates. The results found in the present study contribute to the development of new biocatalysts based on the immobilization of enzymes in nanomaterials.

A simple and low-cost inkjet-printed paper-based electrochemical cell incorporating a molecularly imprinted polymer (MIP) was developed for the selective detection of omeprazole (OPZ). Waterproof paper was used as the substrate for the fabrication of the electrochemical cell by inkjet printing of silver and gold conductive inks, followed by modification of the working electrode with carbon ink. The resulting cell showed good electrochemical performance, with an enhanced voltammetric response towards OPZ compared with conventional screen-printed electrodes. Printing parameters and electrode composition were optimised, and the morphology was confirmed by scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS). The functional monomer was rationally selected through computational studies based on BSSE-corrected binding energies, avoiding a trial-and-error approach. The recognition element was then obtained by electropolymerisation of p-aminobenzene sulfonic acid in the presence of OPZ, forming an imprinted polymer directly on the electrode surface. Under optimised conditions, the sensor exhibited a linear response between 0.50 and 50 µM, with limits of detection and quantification of 0.14 µM and 0.47 µM, respectively, and good selectivity against structurally related compounds. The cell was successfully applied to spiked river and well water samples, without any pre-treatment, achieving recoveries ranging from 89% to 112%. Combining simple fabrication, low cost and portability, this inkjet-printed paper-based MIP sensor shows strong potential for the on-site monitoring of pharmaceutical contaminants.

Maize produces pre-formed secondary metabolites with antimicrobial and invertebrate anti-herbivory properties. Fusarium verticillioides, a mycotoxigenic maize pathogen, detoxifies the maize phytoanticipin 2-benzoxazolinone (BOA) through gene action of FvMBL1, which encodes a putative metallo-β-lactamase within the FDB1 gene cluster. This cluster is also present in other maize-colonizing fungi, including the pathogen Colletotrichum graminicola and, as reported here, the protective endophyte Sarocladium zeae, which exhibits antagonistic activity against mycotoxigenic fungi. The role of the FvMBL1 ortholog in S. zeae, designated SzMBL1, was assessed by gene deletion and add-back complementation. Deletion of SzMBL1 rendered S. zeae incapable of degrading BOA and therefore unable to grow on BOA-amended medium, whereas complemented add-back strains restored BOA tolerance and, in some cases, enhanced growth at higher concentrations of BOA. Quantitative PCR analyses revealed higher copy numbers and expression levels in all add-back transformants relative to the wild type, consistent with their enhanced growth. HPLC-MS/MS analysis further showed that SzMBL1 hydrolyzes BOA, ultimately resulting in biotransformation into the non-toxic metabolite N-(2-hydroxyphenyl)malonamic acid (HPMA). Together, this study confirms the conserved function of the SzMBL1 gene involved in BOA detoxification. This study suggests that the FDB1 gene cluster, harboring SzMBL1 in S. zeae, was likely transferred through horizontal gene transfer between maize-colonizing fungi and plays a role in the degradation of host-derived secondary metabolites that function as filters limiting which fungi are most frequently associated with maize.IMPORTANCEMaize produces secondary metabolites such as 2-benzoxazolinone (BOA) to help protect against microbes and insects. The pathogenic Fusarium verticillioides carries FvMBL1, a gene within the FDB1 gene cluster that helps break down BOA. This cluster is also present in other maize-associated fungi such as Colletotrichum graminicola and, as reported here, the protective fungus Sarocladium zeae. In our experiments, when a gene similar to FvMBL1, denoted SzMBL1, is deleted in S. zeae, the fungus could no longer break down BOA or grow on BOA-amended medium. Complementation restored BOA tolerance and growth phenotypes. Notably, some S. zeae add-back strains showed increased growth at higher BOA concentrations. Our analysis showed that these hypertolerant complemented strains had multiple, transcriptionally active copies of SzMBL1. The study highlights the role of SzMBL1 in BOA detoxification, thereby aiding fungal tolerance to antimicrobial compounds, and supports the hypothesis that the FDB1 gene cluster can be horizontally transferred among maize-colonizing fungi.

Gold(I) vinylidenes are powerful yet typically fleeting electrophiles whose synthetic exploitation has, until now, relied largely on two strategies: 1,2-migration of a halide or silyl group from a gold-activated alkyne, or dual gold catalysis of diyne substrates. Here, we introduce a ligand-enabled catalytic entry to these reactive intermediates based on a prior stoichiometric approach reported by Fürstner. A bifunctional biaryl-2-ylphosphine bearing a remote N,N-dialkylamide group promotes in situ formation of a gold acetylide without an external base; the proton thereby relayed to the amide acts as an internal Brønsted acid that activates a tethered carbonyl, triggering 6-exo cyclization to a putative gold(I) vinylidene. The catalysis converts 2'-ethynylbiaryl-2-yl aldehydes and ketones into synthetically valuable phenanthrene-9-carbaldehydes under mild conditions. In contrast to the observed decarbonylation in the stoichiometric case, the bifunctional ligand is proposed to promote facile protodeauration of an acylgold intermediate, thereby preserving the valuable aldehyde moiety in the products. The transformation accommodates a range of electronic environments and is moderately tolerant of steric demand, underscoring metal-ligand cooperation as a valuable approach to accessing and harnessing gold vinylidenes.

The emergence of antimicrobial resistance represents a major global health challenge, highlighting the urgent need for new therapeutic strategies. Drug repurposing, particularly through the evaluation of FDA-approved compounds with well-established pharmacological profiles, offers a promising avenue. In this study, we performed a phenotypic screening of 31 FDA-approved compounds recently identified as potent Microtubule-Targeting Agents (MTAs) to evaluate their antimicrobial potential. Using agar well diffusion assays, we tested these compounds against selected human pathogenic bacterial and fungal strains. Our results provide the first evidence that several MTAs possess previously unrecognized antimicrobial activities. Of the 31 compounds screened, 18 exhibited inhibitory effects against at least one bacterial or fungal strain, identifying MTAs as potential antimicrobial leads for further investigation. Notably, auranofin, a gold-based antirheumatic agent, consistently displayed antimicrobial activity across all tested strains. Flupentixol dihydrochloride demonstrated selective antifungal inhibition, while mefloquine hydrochloride showed broad-spectrum antibacterial effects. These findings suggest that anti-tubulin activity may contribute to the antifungal effects observed, whereas the possible involvement of FtsZ in the antibacterial activity remains speculative. Further studies, including minimum inhibitory concentration (MIC) determinations, cytotoxicity and selectivity assessments, and mechanistic analyses, are needed to validate these findings and evaluate their potential for antimicrobial repurposing.

Gene-regulatory networks (GRNs) offer a mechanistic framework for dissecting the complex molecular and genetic architecture of coronary artery disease and for identifying new therapeutic opportunities. Here, we present a GRN-driven drug-repurposing strategy that integrates transcriptional signatures induced by silencing key drivers of the human arterial wall foam cell regulator GRN 42 with drug-induced gene expression profiles from the NIH LINCS program. In vitro screening of top candidate compounds validates the computational predictions, identifying candidate foam cell modulators and showing that auranofin, an FDA-approved gold salt used to treat rheumatoid arthritis, effectively reduces foam cell formation. In vivo, auranofin attenuates atherosclerosis and inflammation in both male mice and rabbits. Clinically, auranofin was associated with reduced cardiovascular risk in a retrospective cohort of patients with rheumatoid arthritis. Together, these findings show that a GRN-based drug-repurposing framework, coupled with preclinical and clinical validation, can uncover new therapeutic applications for existing drugs, including auranofin, in coronary artery disease.

Small molecule double-stranded DNA intercalators have significant potential for therapeutic applications. However, screening for and confirming a drug candidate's intercalative behavior remains labor-intensive and costly. To address this, we investigated the sequence and biophysical parameters that affect the performance of electrochemical DNA hairpin sensors for streamlined identification of structural intercalators. These sensors utilize oligonucleotide (oligo) sequences that form hairpins upon intercalator binding. The 3' end of the oligo is modified with alkylthiol linkers for gold electrode surface monolayer self-assembly, while the 5' end carries a methylene blue redox reporter. Hairpin formation enhances electron transfer between methylene blue and the gold electrode, which can be detected via voltammetry. We tested seven hairpin structures varying in stem length and sequence. Our optimal oligo, HP4, features a four-base-pair stem and responds to five DNA intercalators over a broad detection range, with EC50 in close agreement with published affinity (K D) values for these interactions. We further demonstrate HP4's ability to discriminate intercalator binding from a series of minor groove binders through significant differences in signal gain upon incubation. Altogether, our strategy establishes a platform for identifying intercalative compounds that should support the development of DNA-targeting therapeutics.

Salivary extracellular vesicles (sEVs) are accessible candidates for non-invasive biomarker analysis, but their measurement commonly requires multistep, label-dependent workflows. Here a label-free gold extended-gate field-effect transistor (EG-FET) functionalized with an antibody that binds specifically to the Cluster of Differentiation 63 (anti-CD63), was developed for electrical detection of isolated salivary sEVs. Gold gates were modified with an 11-mercaptoundecanoic acid self-assembled monolayer and [1-Ethyl-3-(3-Dimethylaminopropyl) carbodiimide/N-Hydroxysuccinimide] EDC/NHS-mediated antibody immobilization. Binding-induced changes were quantified as the ratio of areas under transfer curves (AUC) acquired before and after incubation. Inter-chip coefficient of variation decreased from 10.5% for bare gates to 1.5% after antibody immobilization. The AUC ratio decreased with sEV concentration across 0.01X-1X (R[2] = 0.980); using the blank mean minus three standard deviations, the estimated detection threshold was approximately 0.1X (about 9.8 × 10[7] particles/mL for the representative stock). Anti-CD63 gates exposed to 0.1X sEVs produced a significant response, whereas Phosphate-Buffered Saline (PBS), Bovine Serum Albumin (BSA), and heterologous-antibody controls remained near baseline. Four independent sessions across a 16-day interval yielded coefficients of variation of 1.4% for PBS and 1.7% for sEV measurements. Response was retained when isolated sEVs were spiked into 0.2X clarified saliva. These proof-of-concept findings support reproducible label-free sEV detection while defining the need for direct whole-saliva interference testing, storage studies, and validation in clinically characterized cohorts.

Arachidonate 15-lipoxygenase (ALOX15) metabolises polyunsaturated fatty acids into lipid mediators that regulate inflammation. Depending on the cell type and substrate, ALOX15 influences both pro-inflammatory and pro-resolving pathways. Its dysregulation is associated with asthma, atherosclerosis, neurodegeneration, ferroptosis, inflammation, oxidative stress and disrupted lipid homeostasis. Therefore, targeting ALOX15 offers a promising strategy for modulating pathological lipid oxidation.

Broad evidence confirms that AR (ICD J30.3) is a chronic disease with high prevalence among children and adults worldwide. Its consequences, both from a public health perspective and at the level of the individual patient, are often underestimated, and the condition is inadequately treated. In addition, AR is often related to comorbidities with additional impact on diseases-severity. Allergen provocation of the easily accessible nasal (nasal allergen provocation test: NPT) or conjunctival mucosa (conjunctival allergen provocation test: CPT) reproduces the allergen-dependent, IgE-mediated allergic reaction under controlled and standardized conditions. These measures are regarded as the diagnostic gold standard, particularly in cases of unclear results in primary allergological tests, enabling clinically relevant identification of the causative allergen. Approved test allergens are critical for ensuring the methodological quality of the NPT. However, their availability is increasingly limited, although adequate quantities of allergen solutions remain indispensable for high-quality nasal allergen testing. Unilateral allergen provocation has been established in the clinical routine for decades in Germany. However, scientific evidence for unilateral application of the test allergen is pending. International position papers differ from this recommendation. The CPT is a simple, safe, and well-tolerated measure. It is primarily used in perennial and, in selected cases, seasonal allergic conjunctivitis. This method is less commonly applied for vernal keratoconjunctivitis and atopic keratoconjunctivitis. It is primarily used to confirm the underlying allergen, particularly when establishing the indication for allergen immunotherapy (AIT). It can also be employed to monitor treatment effects such as e.g., AIT or pharmacotherapy. Although highly sensitive, the procedure of nasal ASA provocation remains a time-consuming procedure. While challenging in children, NPT provides clinically relevant additional diagnostic information on the causes of the allergic symptoms. Allergological diagnostic work-up also includes pulmonary assessment, particularly in patients with established asthma or symptoms suggestive of asthma. Both AAR and PNIF are applied as standard procedures for assessing the outcome of NPT in Germany. In individual cases, other rhinometric methods may also be used.

Olfaction plays a prominent role in the evolution of animal behaviour, facilitating both intraspecies and interspecies communication, and predator-prey interactions. Volatile organic compounds (VOCs) underpin these odours, and their analysis is therefore central to characterising and replicating cues such as pheromones, allelochemicals, and other released signals used to attract, deter, or condition animal responses. Thus, this review aims to introduce the reader to the applications of animal odours in animal management and behavioural control, and to provide a critical, practice-oriented guide to selecting and combining VOC sampling and analytical techniques for animal odour studies. Animal odour profiling typically involves three steps: (1) sampling, (2) transfer to the instrument, and (3) detection/identification of VOCs. Gas chromatography-mass spectrometry (GC-MS) is the gold standard for VOC analysis, and is often paired with headspace techniques, including static headspace, dynamic headspace, and solid-phase microextraction (SPME) to facilitate sampling and transfer of VOCs. In addition, liquid and distillation-based extraction methods are discussed, which remain important for capturing less volatile or matrix-bound components in animal odour and semiochemical research. On the analytical side we discuss GC-MS alongside emerging and complementary techniques, including proton transfer reaction-mass spectrometry (PTR-MS), selected ion flow tube-mass spectrometry (SIFT-MS), gas chromatography-olfactometry (GC-O), and electronic noses, emphasising their respective strengths and limitations for qualitative and quantitative VOC analysis. Collectively, the literature shows that no single sampling or analytical approach is universally optimal. Effective animal odour profiling therefore requires matching extraction and detection strategies to the volatility, stability, and behavioural relevance of target VOCs. Careful consideration should also be given to how these methodological choices influence the odour profiles used in ecological, behavioural, and applied management contexts.

Point-of-care compatible therapeutic drug monitoring requires platforms to rapidly quantify low-abundance small-molecule drugs in complex biofluids while resolving spectral interference from coexisting compounds. Surface-enhanced Raman spectroscopy offers molecular fingerprint specificity, yet its quantitative reliability is often compromised by heterogeneous hotspots, matrix interference, and overlapping drug spectra. Here, we develop an integrated microfluidic SERS platform based on charge-directed 3D Au@Ag plasmonic assemblies and machine learning-assisted spectral decoding. Oppositely charged gold nanopolytopes and silver nanospheres spontaneously assemble into ordered 3D structures with dense interparticle nanogaps. Optimized 1:1 Au@Ag assembly delivers enhanced electromagnetic coupling and stable, reproducible SERS signals. Combined with deuterated methanol as a ratiometric internal standard in microfluidic chips, this system achieves low-volume, fluctuation-corrected standardized drug analysis. Six therapeutic molecules are identified via SERS fingerprints, and binary or ternary mixtures are accurately decoded by 3D-LDA and CNN-RF with AUC > 0.98. Reliable discrimination in rat plasma, artificial sweat, and urine demonstrates excellent matrix tolerance. This work establishes a material microfluidic algorithm-integrated SERS strategy for rapid therapeutic drug analysis, paving the way toward portable intelligent drug monitoring systems.

A portable ozone detection strategy is reported based on derivatization-assisted surface-enhanced Raman spectroscopy (SERS). The strategy integrates active sampling, chemical derivatization, and signal enhancement using high-index gold trisoctahedrons embedded in a porous hydrogel (Au TOHs/hydrogel), assembled into a 48-well microdevice. Ozone is selectively captured by eugenol and converted to formaldehyde, which reacts with 3-methyl-2-benzothiazolinone hydrazone (MBTH) to form a Raman-active azo compound, enabling sensitive detection. The SERS signal at 1270 cm[- 1] exhibited a strong linear correlation with the logarithm of analyte concentration (R[2] = 0.996). The limit of detection under simulated air conditions is 5.80 × 10[- 6] mg/m[3]. The SERS sensor demonstrated high stability, spatial uniformity, and reproducibility (RSD < 10%), along with good selectivity against common volatile organic compounds. Validation with air samples and comparison with ultraviolet-visible UV-vis spectrophotometry further supports its applicability. This cost-effective and user-friendly platform shows potential for point-of-use detection of trace ozone.