Nanoneedles are emerging as a safe and scalable strategy for the genetic modification of primary human cells. However, a limited understanding of how interactions at the biointerface lead to functional gene expression continues to hinder clinical translation. While direct membrane penetration, permeabilization, and endocytosis have been proposed as intracellular delivery avenues, the mechanistic connection between delivery and successful transfection remains unclear. Here, we identify caveolae-mediated endocytosis, dependent on Caveolin-1, as a key mechanism enabling nanoneedle transfection. By selectively modulating Caveolin-1 expression in primary human regulatory T cells and MG63 cells and investigating endolysosomal processing, we show that although nucleic acids can be efficiently delivered in the absence of Caveolin-1, gene expression occurs only when caveolar endocytosis is present. These findings reveal a mechanistic basis and establish a broader design principle for nanoneedle transfection: interfacing must be accompanied by the engagement of permissive cellular trafficking pathways to achieve gene expression.

An AIE-active triphenylethylene probe for ultrasensitive detection of peroxynitrite and imaging in living cells.

Cationic peptides, particularly those rich in arginine and/or lysine residues, are usually promising antimicrobial agents effective at low concentrations in laboratory settings. However, their applicability in pharmaceutics and biotechnology is currently limited due to their susceptibility to biological enzymatic processes and (in some cases) toxicity to host cells. To address this, we screened eight linear arginine-rich peptides for their haemolytic properties and antimicrobial activity using a set of computational and experimental assays. Inspired by our previous results on R4F4, we then designed three modified peptides based on an R4F4 backbone, R4F4-C16, D-R4F4 and cyclic R4F4, and one based on R4 (R4-C16). Amongst the tested linear peptides containing only natural amino acids, R4F4 exhibited the strongest antibacterial activity; however, this effect was reduced in the presence of human serum and trypsin. Conversely, our study demonstrated that cyclization and substitution to its d-amino acid enantiomer significantly enhanced stability and activity of R4F4, whilst in the presence of proteases. As revealed by fluorescence imaging, microscopy RNA sequencing analysis, the mode of action involves complex and dynamic events. This multifaceted antimicrobial mechanism integrates alterations in membrane permeability, modulation of intracellular reactive oxygen species levels and changes in transcriptomic signature profiles. At the molecular level, notable changes were observed in the bacterial expression of genes associated with metabolic pathways and biological processes. Furthermore, R4F4-derived peptides showed substantial antibiofilm activity in preventing the formation and disruption of mature biofilms, together with good cytocompatibility, highlighting the potential for clinical applicability. In conclusion, this study emphasizes the importance of optimizing the stability of peptide-based antimicrobials, particularly those rich in arginine, and highlights the advantages of incorporating d-amino acids and cyclization for enhanced performance. This information will prove useful in the future design of antimicrobial peptides. In addition, the molecular perspective on peptide-induced gene expression changes, as identified by RNA-seq, broadens our understanding of antimicrobial peptides' activities and provides a clearer picture of their versatile mechanisms.

ABSTRACT Background Mitochondria and lysosomes are pivotal in dictating cell survival or death outcomes. While mitochondrial damage and ROS production are key events in mitochondrial cell death, lysosome membrane permeabilization and cathepsin B release mark lysosomal cell death. We aimed to generate a live-cell approach to concurrently monitor mitochondrial redox alterations and lysosomal permeabilization. This would provide mechanistic insight into their dynamic interplay during cell death and enable the discovery of organelle-specific death inducers. Methods A dual cell sensor, stably expressing tdTomato-CathepsinB and mitochondria-targeted redox GFP (mt-roGFP), was successfully engineered, and simultaneous imaging of both events by real-time confocal imaging was carried out with selected drugs. Results This platform faithfully reported the chronological sequence of organelle-specific events with the progression of cell death, with good temporal and spatial resolution at the single-cell level. Moreover, we have identified and categorised potential lead compounds that predominantly induce lysosomal cell death or mitochondrial cell death, as well as a subset that elicit both events concomitantly. Conclusion The study provided evidence that both organelles contribute to cell death in a context-dependent manner, and the temporal analysis of both events is critical in understanding unique organelle-centred cell death.

Antiviral substances are considered emerging contaminants. Once released into the environment, they may affect organisms through complex and often still-unknown mechanisms. This study focuses on a class of antiviral substances with potential use in treating COVID-19 patients, aiming to identify specific structural characteristics that significantly contribute to their ecotoxicity. An empirical approach called quantitative structure–activity relationship (QSAR) was used for this purpose. The study examined 13 antiviral substances: atazanavir, daclatasvir, darunavir, emtricitabine, favipiravir, lopinavir, nirmatrelvir, oseltamivir, remdesivir, ribavirin, ritonavir, and sofosbuvir. The ecotoxicity of these antivirals was assessed using three tests: the Aliivibrio fischeri test, the Chlorella sp. test, and the Pseudomonas putida test. These three microorganisms represent different trophic levels in aquatic and soil ecosystems. Ecotoxicity was expressed as EC20 and EC50, and these values served as the dependent variables in the QSAR models. A large set of numerical descriptors calculated from the molecular structures of the antivirals was used as an independent variable. EC20-based QSAR models offer insight into the effects of antivirals under sub-lethal exposure conditions. The results indicated that sub-lethal exposure in Aliivibrio fischeri was associated with favorable electronic properties and compact structures that promote cellular accumulation, while long-range fragments reduced toxicity. In Chlorella sp., sub-lethal exposure was driven by optimal molecular size, chain length, and specific electronic groups enabling cell penetration and biochemical inhibition. For sub-lethal exposure in P. putida, lipophilicity and reactive group geometry enhanced toxicity, while high short-range polarity mitigated it by limiting membrane permeability. Acute toxicity patterns showed similar trade-offs: strong electronic reactivity increased potency, but steric bulk, long-range polarity, or unfavorable mass distribution frequently restricted bioavailability and reduced toxic effects. Overall, the models demonstrated that antiviral toxicity results from a balance of electronic activity, structural accessibility, and physicochemical constraints, providing a mechanistic basis for predicting the environmental risk of selected antiviral substances.

Butyl Rubber (IIR) and Brominated Butyl Rubber (BIIR) are potential candidates for the sealing solutions in the lithium-ion battery area. In this work, the electrolyte resistance of IIR- and BIIR-based membranes under various thermo-oxidative conditions is characterized by the evolution of macromolecular structures, mechanical properties, and fluid transport performance. The changes in mechanical properties of the BIIR are more evident than those of IIR after aging under various conditions. FTIR analysis reveals that -Br groups induce aberrant cross-linking and oxidation of BIIR, generating carbonyl groups at 1710 cm-1. Transport kinetics indicates that BIIR exhibits a higher diffusion coefficient and permeability coefficient, while IIR maintains a lower uptake ratio and dimensional stability within the range of 45-80 °C. The electrolyte swelling experiment shows that the residual electrolyte crystal powder on the surface of IIR and BIIR samples increases with the temperature. The permeation activation energies (EP) accounting for the electrolyte resistance are fitted by the Arrhenius equation for IIR and BIIR samples, being in the range of 177.4-208.8 kJ mol-1 and 170.8-203.0 kJ mol-1, respectively. Theoretical support can be obtained from this work for the selection of sealing membrane materials and prediction of service life in the lithium-ion battery area under enhanced thermal-oxidative conditions.

Lactobionic acid (LBA) has demonstrated antibacterial activities against multiple foodborne bacteria; however, few studies have reported on its effect against Cronobacter sakazakii. In this study, the antibacterial activity and mode of LBA against C. sakazakii were explored. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of LBA against C. sakazakii were 12.5 and 25 mg/mL, respectively. LBA exhibited bacteriostatic activity at sub-MIC and bactericidal activity at concentrations ≥ MIC. Alkaline phosphatase (AKP) activity, cell outer membrane (OM) permeability, protein leakage, and gel electrophoresis results suggested that LBA increased the permeability of the cell wall and OM, leading to intracellular protein leakage and a decrease in protein contents and activity, indicating LBA damage to the cell wall and membrane. Among these, the rapid AKP activity surge reached 4.37 U/gprot at 2 MIC, and the OM permeability dramatically increased up to 10 min and stabilized after 30 min. Microscopic observations confirm the disruption to the cell wall and membrane, further showing that LBA disrupted the integrity of the cell wall and membrane. Moreover, LBA disturbs normal cellular functions by binding to deoxyribonucleic acid (DNA), as reflected by the competitive binding assay. Overall, LBA possesses potential multiple applications in the food industry due to its natural and antibacterial properties.

Colistin is considered one of the last-resort antibiotics for treating infections caused by multidrug-resistant (MDR) Gram-negative bacteria. However, the emergence and dissemination of mobile colistin resistance gene, mcr, have severely compromised the clinical utility of colistin. Combination therapy has emerged as a promising strategy to restore and enhance antibiotic efficacy against such bacterial infections. In this study, we identified broxyquinoline (BRO), an antiprotozoal compound, as a potent colistin adjuvant that significantly enhanced colistin activity against both colistin-susceptible and colistin-resistant Gram-negative bacteria by markedly reducing the minimum inhibitory concentration. Mechanistically, BRO disrupts bacterial membrane integrity, increases membrane permeability and fluidity, collapses the proton motive force, induces reactive oxygen species (ROS) accumulation, and depletes intracellular ATP, collectively disturbing bacterial homeostasis. Additionally, BRO exhibited high-affinity binding to lipopolysaccharide (LPS) and attenuated subsequent LPS-induced inflammatory responses in host cells. In murine thigh and lung infection models, the BRO-colistin combination restored colistin efficacy in vivo, evidenced by significantly reduced bacterial loads. In the lung infection model, this combination further improved survival, alleviated pulmonary pathological damage, and reduced the levels of pro-inflammatory cytokines (TNF-α, IL-1β) in bronchoalveolar lavage fluid. Collectively, these findings support the BRO-colistin combination as a promising therapeutic strategy to overcome colistin resistance and combat MDR Gram-negative infections.

Site-specific propynylation modification of apigeninidin enhances anti-cervical cancer activity by targeting PARP-1.

Tunable gas permeability and interfacial engineering of poly(propylene carbonate)/bamboo powder composite films for sustainable packaging.

Oxygen permeability and stability in the entropy-stabilized Co-based perovskite oxygen permeable membranes

ABSTRACTDysregulation of autophagy and lysosomal function is central to Parkinson's disease (PD), yet the upstream mechanisms leading to lysosomal failure remain unclear. Across primary mouse cortical neurons, MT‐3 deficient primary mouse astrocytes, human iPSC‐derived midbrain dopaminergic neurons, and Rho0 CHO cells lacking mitochondrial respiration, we investigated how mitochondrial stress perturbs zinc (Zn2+) homeostasis and lysosomal integrity. We identify intracellular zinc as a critical mediator linking mitochondrial dysfunction to lysosomal membrane permeabilization (LMP) and neuronal death. Inhibition of mitochondrial complex I by 1‐methyl‐4‐phenylpyridinium (MPP+) elevated reactive oxygen species (ROS) and intracellular zinc, jointly driving LMP. Blocking either ROS or zinc markedly attenuated lysosomal damage and cell death, demonstrating that both act upstream of LMP. To define zinc regulation, we examined metallothionein‐3 (MT‐3), a brain‐enriched zinc‐binding protein. MT‐3‐deficient astrocytes were more vulnerable to MPP+ and zinc overload (ZnCl2) but paradoxically resistant to hydrogen peroxide (H2O2), suggesting that MT‐3 buffers cytosolic zinc during mitochondrial injury or extracellular zinc influx yet can release bound zinc under oxidative conditions. Using Rho0 cells, we show that MPP+ toxicity depends on mitochondrial ROS, as loss of mitochondrial function nearly abolished cell death. However, Rho0 cells were highly sensitive to ZnCl2 and H2O2 and exhibited markedly reduced lysosomal abundance, indicating limited capacity to sequester zinc and increased susceptibility to zinc‐mediated injury. These findings support a coordinated system in which lysosomes and zinc‐binding proteins maintain zinc homeostasis. When cytosolic zinc rises, its accumulation within lysosomes induces LMP and accelerates cell death. Collectively, our results identify intracellular zinc as an upstream trigger of lysosomal dysfunction and neurodegeneration. Zinc‐mediated LMP provides a mechanistic link between mitochondrial injury, impaired autophagic flux, and α‐synuclein pathology in PD. Enhancing zinc homeostasis and lysosomal resilience may offer promising therapeutic strategies.

Engineering lysosomal collapse for cancer therapy: From mechanistic insights to nanotherapeutic innovations.

ZIP14 upregulation leads to ferroptosis and lysosomal dysfunction through intracellular iron overload and induces myocardial ischemia/reperfusion injury in mouse hearts.

Real-world aged microplastics exacerbate antibiotic resistance genes dissemination in anaerobic sludge digestion via enhancing microbial metabolite communication-driven pilus conjugative transfer.

Discovery of novel pyrrolidine-2,5-dione scaffold PICK1 PDZ inhibitors as anti-ischemic stroke agents.

New-generation advanced nano-PROTACs as potential therapeutic agents in cancer therapy.

Responses and regulatory mechanisms of soil microbiome and antibiotic resistome to carbendazim and ZnO nanoparticles.

A review of ammonia toxicity on aquatic organisms: Species-specific responses, microbial shifts, and environmental interactions.

Comamonas aquatica, an emerging nosocomial pathogen, poses significant clinical challenges through biofilm-mediated antimicrobial resistance. This study investigated the efficacy of cuminaldehyde combined with tetracycline against C. aquatica biofilms using an integrated approach. In silico predictions (PASS online, SwissADME, PROTOX 3.0, OSIRIS) indicated that cuminaldehyde exhibited favorable oral bioavailability with acceptable toxicity profiles, while tetracycline showed limited oral absorption due to molecular size and polarity constraints. Experimentally, individual minimum inhibitory concentrations (MICs) were determined as 300 μg/mL for cuminaldehyde and 0.2 μg/mL for tetracycline. The fractional inhibitory concentration index (FICI) of 0.66 demonstrated additive interactions between the compounds (cuminaldehyde and tetracycline). The result indicated that the combinatorial application of compounds exhibited enhanced antimicrobial potential against the test organism. Furthermore, co-application of cuminaldehyde and tetracycline was found to show increased antibiofilm potential against the same organism. The result showed that the biofilm inhibition under the influence of the combinatorial application could be attributed to the enhancement of bacterial cell membrane permeability and accumulation of intracellular reactive oxygen species. In a nutshell, the findings of this study highlight a promising strategy of using combinatorial therapy involving cuminaldehyde-tetracycline for dealing with biofilm-associated infections caused by C. aquatica.