DNAzymes are frequently adopted in biosensors for their sensitivity and specificity in detecting and managing food safety concerns. Depending on the DNAzyme variant, they can function as recognition elements or signal reporters in biosensors. This review comprehensively evaluates the incorporation of DNAzymes in optical and electrochemical biosensing. It provides a comprehensive overview and understanding of the development of DNAzyme-based biosensors for food safety detection in the food industry. The brief introduction, development history, and immobilization strategy of DNAzyme biosensors in food were highlighted to improve the working of DNAzyme-based biosensors. The detection process of DNAzyme-based biosensors is formulated, emphasizing the catalytic cleavage of the analytes and the subsequent signal amplification methods used to improve sensitivity. The emerging optical and electrochemical sensors based on DNAzyme are also classified according to their sensing mechanism towards food safety. Lastly, the hurdles and future research expectations of DNAzyme biosensors for food safety detection are highlighted to improve selectivity, specificity, and potential for integration with other technologies. This article provides a comprehensive review of DNAzyme-based biosensors for detecting food safety, highlighting the potential for further development in this field.

Correction for ‘Artificial intelligence-driven dynamic regulation for high-efficiency gentamicin C1a production’ by Feng Xu et al. , Green Chem. , 2025, 27 , 13436–13454, https://doi.org/10.1039/D5GC02507A.

The clinical importance of gentamicin C1a as a broad-spectrum aminoglycoside antibiotic underscores the need for efficient biomanufacturing strategies. In this study, we developed a systematic engineering framework to enhance gentamicin C1a production. First, a genome-scale metabolic model (iFX1172) was reconstructed to pinpoint critical bottlenecks in both regulatory and biosynthetic pathways. Guided by model predictions and experimental validation, we identified genC, metK, and BldD as synergistic targets. Coordinated overexpression of these genes increased gentamicin C1a titers to 198.1 mg/L, representing a 34.3% improvement over the parental strain, and also enhanced the titers of other aminoglycoside antibiotics by up to 1.6-fold, demonstrating the universality of the strategy. Metabolic flux analysis and targeted metabolomics revealed that redox homeostasis and ATP availability are pivotal for biosynthesis. Finally, process optimization in a fed-batch bioreactor using a Bayesian framework, coupled with in situ resin adsorption, yielded 964.1 mg/L gentamicin C1a with a yield of 24.1 mg/g glucose and a productivity of 6.7 mg/L/h.

Coenzyme Q10 fermentation by Rhodobacter sphaeroides (R. sphaeroides) is highly aerobic. However, its biosynthesis and accumulation are paradoxically induced under hypoxic conditions. While oxygen serves as both an induction signal and a key precursor, the mechanisms underlying Coenzyme Q10 accumulation under hypoxia remain elusive, posing a significant bottleneck for yield improvement. This study systematically elucidated these mechanisms through transcriptomic analysis. Results revealed that metabolic pathways for precursor and Coenzyme Q10 biosynthesis were, surprisingly, downregulated under hypoxia. Conversely, genes inhibiting morphological remodeling were downregulated, while those involved in cell membrane biosynthesis were upregulated. Furthermore, a significant positive correlation was observed between cellular morphology and Coenzyme Q10 yield. Specifically, under varying hypoxic conditions, cell morphology and Coenzyme Q10 yield also exhibited strong correlations, with higher yields were associated with larger cell volumes. These findings suggest that R. sphaeroides enhances Coenzyme Q10 accumulation primarily by expanding the cell volume and membrane surface area rather than upregulating biosynthetic pathways. Leveraging this insight, the addition of unsaturated fatty acids further increasing the Coenzyme Q10 yield by 18.9%. This study provides a novel strategy for enhancing Coenzyme Q10 production through morphological engineering and process optimization.

Promoting rapid wound healing is essential for preventing hypertrophic scars. Exosome-based therapies represent a promising approach for improving healing outcomes, but they encounter significant challenges: 1) the isolation of exosomes requires complex, time-consuming, and costly procedures; 2) the stability of exosomes is limited due to the pH-and temperature-sensitive nature of their lipid bilayer. Hydrogels, known for their hydrophilic and porous architectures, offer an attractive solution by enabling the isolation of exosomes from macromolecular impurities while simultaneously protecting them from degradation. In this study, we designed a bilayer polysaccharide hydrogel where pullulan (PL) and calcium ions (Ca) form the inner core, while sodium alginate (SA) creates an effective isolating gel in the exosome-containing precursor solution. By integrating freeze-drying with ultrasonic-assisted dissolution, the approach effectively achieves both the isolation and controlled release of exosomes. The results show that the PL + Ca + SA hydrogel exhibits superior gelation stability compared to other formulations. During isolation, the PL + Ca + SA hydrogel efficiently enriched exosomes while maintaining their characteristic "teacup-shaped" morphology and protein markers. The isolation yield (87.2%) was comparable to that obtained by ultracentrifugation, with no significant differences in particle number or impurities. Furthermore, ultrasonic-assisted dissolution enabled sustained exosome release for at least 7 days, as predicted by the Higuchi diffusion model. Functionally, the EXO@PL + Ca + SA hydrogel enhanced fibroblast proliferation and viability for up to one week, changing the ROS content (decreasing 92%) and the COL-I/COL-III ratio to 0.58 of mouse wound model. Furthermore, transcriptomic analysis revealed significant alterations in related pathways involving key genes. Overall, this bilayer hydrogel offers an integrated platform for exosome isolation and controlled delivery, providing preliminary experimental evidence to support further translational research.

In this study, Se-doped CeO2@Fe3O4 nanoparticles (NPs) were synthesized and applied to a Carthamus tinctorius (safflower) cell suspension culture using liquid medium (B5). The application of these NPs at various levels (0, 5, 10, 15 and 20 mg L-1) was studied for its effects on cell growth, physio-biochemical traits and antioxidative activities. The addition of NPs to the culture media significantly improved the cell biomass, antioxidant potential and phenolic contents. The addition of NPs at the rate of 15 mg L-1 (T3 treatment group) significantly improved the dry biomass of cells (128.72%), total chlorophyll contents (76.02%), and reduced levels of hydrogen peroxide (5.15%) and reactive oxygen (26.51%) compared to the control group (0 mg L-1). Furthermore, this study identified 29 differentially expressed genes (DEGs) in the jasmonate signalling pathway. Notably, only the two DEGs from the MYC2 family showed mixed expression at different time points (6 h, 24 h, 48 h, and 72 h) following treatment with Se-doped CeO2@Fe3O4 NPs. In conclusion, these findings demonstrate that this approach is effective, adaptable, biocompatible, and cost-efficient, offering a promising strategy for enhancing the production of antioxidant and bioactive metabolites in industrial-scale safflower cultivation.

Organoid-based therapy, as an approach in regenerative medicine, offers new options for previously untreatable diseases. However, the limitations of organoid transplantation, such as immunogenicity, tumorigenic potential, and ethical issues, restrict its clinical translation. For the first time, a novel three-dimension matrigel-free suspension culture system is developed to generate liver ductal organoid-derived extracellular vesicles (3D OEVs) for primary sclerosing cholangitis (PSC) treatment. The results show that the developed suspension culture provides a more favorable mechanical microenvironment, which significantly enhances the functional maturation of liver ductal organoids and their extracellular vesicles. This improvement is facilitated by a multidimensional regulatory network that encompasses the PI3K-AKT and RAP1 pathways. Moreover, 3D OEVs significantly attenuate hepatic inflammation and fibrosis because they can remodel the immune microenvironment, polarizing more macrophages into the anti-inflammatory M2 phenotypic macrophages. Furthermore, miR-1299 enriched in 3D OEVs is identified as the core effector molecule for macrophage reprogramming, which was confirmed by both in vivo and in vitro experiments to effectively repair biliary damage through suppressing the EGR1/FOS/RAS signaling axis. As a result, a scalable platform for the production of therapeutic OEVs was successfully developed in a 50 mL bioreactor utilizing a matrigel-free methodology, thus offering a bioactive material for liver regeneration.

BACKGROUND:Synthesis of novel enzyme inhibitors and antioxidants is active research in pharmaceuticals sciences. Novel compounds with potential therapeutic value can be synthesized to help the body combat oxidative-stress related diseases. OBJECTIVES:This study comprised synthesis of 4-(3-bromophenyl)-[2,2'-bipyridine]-6-carboxylic acid, its complexation with Fe(II), Cu(II) and Ni(II), and the evaluation of these compounds as lipoxygenase inhibitors and antioxidant agents. METHODS:The ligand and its complexes were characterized using NMR, IR, UV-Vis and mass spectroscopies. The synthesized compounds were also screened to evaluate their antioxidant and lipoxygenase properties. Two sample t-test and ANOVA were used to evaluate the significance of biological activity of the complexes. RESULTS:The ligand acted as a tridentate donor forming ML2 complexes. Cu(II) and Ni(II) complexes showed significant lipoxygenase inhibition with the respective IC50 values of 50.6 and 59.8 µM, while antioxidant activity was largely attributed to the ligand itself with IC50 value of 56.4 µM. CONCLUSION:The study demonstrated that Cu(II) and Ni(II) complexes of the synthesized ligand were promising candidates for lipoxygenase inhibition, while the free ligand exhibited significant antioxidant activity.

Intratumoral redox homeostasis is often implicated in the development of multidrug resistance (MDR), which compromises the oxidative cytotoxicity of anthracycline chemotherapeutics such as doxorubicin (DOX). To overcome the redox homeostasis-driven MDR, we analyzed the transcriptomes of 216 breast cancer patients, revealing a noteworthy association between the TRPA1 (a regulator of redox homeostasis) overexpression and drug resistance. Moreover, a smart magnetic nanozyme, HADAF (HA@Anti-TRPA1@DOX@Au@Fe3O4), was rationally designed to target TRPA1 and reverse MDR. HADAF nanozyme integrates Au@Fe3O4 nanosheets with DOX, antisense oligonucleotides of TRPA1 and hyaluronic acid (HA). Notably, the dynamic regulation of HADAF is triggered spatiotemporally by a controllable low-frequency vibrational magnetic field (VMF) and near-infrared (NIR) laser irradiation. In vitro and in vivo experiments showed that HADAF effectively inhibited the TRPA1/PI3K/mTOR/MCL-1 signaling pathway, promoting apoptosis and ferroptosis, and achieving a 90% inhibition rate in MCF-7/ADR cells. As a result of this engineered therapeutic approach, the combination therapy reduced the IC50 of DOX by 87.7-fold compared to free DOX. In summary, this multimodal nanozyme provides a novel strategy to target redox homeostasis and overcome MDR in cancer therapy.

Chinese hamster ovary (CHO) cells are the primary platform for therapeutic antibody production. Although histone deacetylase inhibitors such as sodium butyrate (NaBu) can enhance recombinant expression, their growth-inhibitory and cytotoxic effects often limit volumetric productivity. Here, a structure-guided docking strategy was applied to prioritize NaBu-derived small-molecule additives (SMAs) for experimental screening in CHO antibody-producing cell lines. A lead combination (D1 +D4) increased volumetric antibody titers by approximately 2-3 fold while maintaining high cell viability (>95%) under the tested conditions. Cell-based analyses indicated reduced apoptotic markers and altered cell-cycle distributions in D1 +D4-treated cultures relative to NaBu. Metabolite profiling further revealed reduced by-product accumulation, including lactate and ammonia, together with a distinct intracellular energy and redox state compared with control cultures. Importantly, N-glycan profiles and charge variants of the produced antibodies remained comparable between control and D1 +D4-treated cultures. The productivity benefit of D1 +D4 was further maintained in bench-scale stirred-tank and wave bioreactor systems, supporting process relevance. Together, these results demonstrate that structure-guided NaBu analog screening can identify practical additives that enhance CHO antibody productivity with minimal impact on product quality.

This study investigates the enzyme inhibition and antioxidant activities of various nanoparticles loaded with three isatin thiazole derivatives. For drug encapsulation, two different nanoparticle systems were examined: polysaccharide-based nanoparticles, including alginate-chitosan nanoparticles (ACN) and gum-chitosan nanoparticles (GCN), and polymer-lipid hybrid nanoparticles based on the lipid soya lecithin in combination with either sodium alginate (PLHN-A) or gum acacia (PLHN-G). The biological activities of the drug-loaded nanoparticles were evaluated against key targets, including antioxidant activity, urease, lipoxygenase, and butyrylcholinesterase. The results were expressed as mean +/- SEM, using eserine, butylated hydroxyanisole (BHA), and thiourea as reference standards. Among the tested systems, PLHN-A loaded with derivative-3 (D-3) exhibited the highest antioxidant activity (30.2 +/- 0.45). Alginate-chitosan nanoparticles loaded with derivative-3 (ACN-D-3) demonstrated notable anti-urease activity (21.2 +/- 0.12), while derivative-1(D-1) encapsulated in the same nanoparticle system (ACN-D-1) exhibited significant lipoxygenase inhibition (13.4 +/- 0.48). Gum-chitosan nanoparticles loaded with derivative-1 (GCN-D-1) demonstrated strong butyrylcholinesterase inhibition (IC50 = 21.6 +/- 0.17 & micro;M). Overall, ACN and PLHN-A with all three derivatives (D-1, D(2 )and D-3) emerged as promising candidates due to their excellent antioxidant and enzyme inhibition properties, underscoring the effect of nanoparticle composition on their biological activity and therapeutic potential.

BACKGROUND:This study presents a simple and eco-friendly green synthesis approach for the preparation of amino acid-assisted zinc oxide nanostructures (ZnONSs). OBJECTIVES:The objectives of synthesizing amino acid-assisted ZnONSs were to evaluate their antibacterial applications. METHODS:Among the amino acids selected are Arginine, aspartic acid, cystine, and lysine. These amino acids were used as capping and stabilizing agents to tailor the structural and surface properties of ZnONSs. The synthesized nanostructures were characterized using X-ray diffraction (XRD), UV-Visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDS) and disk diffusion method for antibacterial activity analysis. RESULTS:XRD analysis confirmed the formation of ZnO with altered crystallinity due to amino acid incorporation, while UV-Vis spectroscopy verified successful synthesis of ZnONSs. FTIR spectra demonstrated effective surface functionalization by amino acid-related functional groups, and SEM revealed amino acid-dependent morphological variations, including rod- and flower-like structures with increased surface area. EDS further verified the predominance of Zn in the samples. The disk diffusion study confirmed effective antibacterial activity against Staphylococcus aureus (Gram-positive) and Escherichia coli (Gram-negative), with performance varying according to the amino acid used during synthesis. Among the samples, arginine-assisted ZnONSs demonstrated superior antibacterial efficacy, likely due to enhanced surface functionalization and electrostatic attraction with negatively charged bacterial cell membranes. CONCLUSION:This work highlights amino acid-assisted green synthesis as a promising route for developing eco-friendly ZnO-based antibacterial nanomaterials.

The declining efficacy of azithromycin-the primary oral treatment for typhoid fever in XDR-endemic regions-highlights the urgency against drug-resistant Salmonella Typhi. In response, we developed two ƞ6-arene ruthenium(II) series (9-20 and 37-45) by integrating natural product-based antimicrobial motifs with the DNA-intercalating/ROS-generating features of Ru-scaffolds. Among these, complex 11 and 16 exhibited potent activity against MDR and XDR S. Typhi, with lead complex 16 showing MIC90 as low as 0.06 μg/mL and MBC values significantly superior to azithromycin. Time-kill kinetics showed rapid, concentration-dependent bactericidal effect (≥3-log10 CFU reductions within 8 h), while negligible hemolysis (<2 % at 32 μg/mL) confirmed biocompatibility. Mechanistic studies revealed dual action-membrane disruption and DNA replication inhibition-supported by delayed invA amplification (increased Cq at 1 × /2 × MIC90) and strong binding to DNA polymerase III (docking S-scores: -8.01). Favorable ADMET profiling, including high intestinal absorption (TPSA ∼70 Å2) and no CYP450 inhibition, further highlights the potential of these complexes as new option against untreatable typhoid.

Breast cancer poses a significant health threat to women, as traditional chemotherapy often fails due to multidrug resistance and tumor adaptability. Moreover, the anti-apoptotic nature of these tumors limits the effectiveness of conventional treatments. Current therapeutic approaches, including photodynamic therapy (PDT), face limitations in clinical application due to poor water solubility of photosensitizers and challenges in targeted delivery. This study aims to develop a multifunctional nanosystem that integrates PDT with gene therapy to enhance breast cancer treatment efficacy. Firstly, Fe3O4 hollow nanospheres were prepared to carry Ce6 drug to kill breast cancer cells via chemodynamic therapy and PDT, respectively. To overcome the drawback of poor water solubility of PDT, HA/G3139/Ce6@Fe3O4 nanosystem was synthesized, which combines the benefits of G3139 antisense oligonucleotides for gene therapy and hyaluronic acid for encapsulation and targeting CD44 receptor. The developed novel nanosystem exhibits excellent biocompatibility and pH responsiveness, significantly enhancing its cancer cell-killing capability under light exposure. When the therapeutic system enters cancer cells and decomposes in the acidic environment, it releases Ce6 and G3139. On one hand, the photosensitizer Ce6 generates cytotoxic ROS upon irradiation, killing cancer cells; on the other hand, G3139 binds to the anti-apoptotic gene BCL-2 in cancer cells, downregulating the protein and inhibiting tumor proliferation. The novel nanosystem demonstrates synergistic anti-cancer effects by combining PDT, gene therapy, and chemodynamic therapy, leading to enhanced apoptosis in breast cancer cells. In conclusion, this approach offers a promising strategy for more effective and targeted breast cancer treatment.

Biothiols (Cys/Hcy/GSH) have emerged as prime biomarker of cancer benefiting in the recent research playing a source of detection via fluorescence imaging. Pre-eminent present-day tools for biothiol detection are fluorescent probes that overcome the limitations of conventional detection methods by dispensing high selectivity, admiring sensitivity, low background-to-noise ratio, low anti-interference, and quick low-cost detection. Here, we report a freshly synthesized red-emitting BD-MSN fluorescent scaffold for the efficient in vivo detection of endogenous biothiols (Cys/Hcy/GSH) in living HeLa cancer cells and zebrafish larvae possessing the aforementioned features along with high water solubility, excellent biocompatibility, and low cytotoxicity. The reported probe has been amalgamated by the courtesy of nucleophilic substitution reactions that ensure biothiols-influenced hypersensitive detachment of functional group (BD) from the fluorophore (MSN-OH) offering strong red fluorescence emission at 635 nm. Moreover, successful in vitro and in vivo trials prove that BD-MSN can potentially be utilized for cancer detection in living organisms.