Relapsed and refractory cancers effectively overcome diverse modalities of cancer treatment, whose principal targets are nucleic acids and proteins. The plasma membranes of cancer cells represent an alternative and underutilized target, with the potential for membrane lysis to induce rapid, pro-inflammatory cell death that circumvents the challenges of intratumor heterogeneity and immune evasion. Here, we repurposed StAMP51.2, a stapled magainin 2 peptide previously optimized for selective membrane lysis of gram-negative bacteria, to target cancer cell membranes. The PRISM assay, a high throughput cancer cytotoxicity screen, revealed cancer cells most vulnerable to StAMP51.2 and biomarkers of susceptibility, specifically reduced cholesteryl esters and elevated triacylglycerols. This signature was validated in pairs of sensitive (OCI-AML3, THP-1) and resistant (K562, KSM-11) leukemia cell lines, with their differential responses correlated to distinct lipidomic profiles. Susceptibility of OCI-AML3 cells in culture extended to the in vivo context, where StAMP51.2 suppressed leukemic growth in orthotopic and intraperitoneal models. To further characterize the mechanism of action, StAMP51.2-resistant OCI-AML3 cells were generated, which required four months of low-level exposure. Strikingly, drug-resistant OCI-AML3 cells recapitulated the lipidomic phenotype of naturally resistant K562 cells. Transcriptomic analyses further revealed that lipid reprogramming was accompanied by pervasive downregulation of inflammatory signaling. Thus, in advancing StAMP51.2 as an oncolytic prototype, this study uncovered an immune regulatory axis that links membrane integrity to inflammatory signaling.

Hepatocellular carcinoma (HCC) represents one of the most fatal cancers worldwide, characterized by high mortality rates and poor prognosis. ABT-737, a small-molecule antagonist of the Bcl-2 family, has shown promise as an anticancer agent. However, the potential of ABT-737 to induce PANoptosis-a unique inflammatory programmed cell death pathway encompassing apoptosis, necroptosis, and pyroptosis-remains unexplored in HCC. This study aimed to investigate the potential of ABT-737 to induce PANoptosis in hepatocellular carcinoma cells and elucidate the underlying mechanisms governing its effects on proliferation, migration, and invasion. Two human HCC cell lines (SK-HEP-1 and BEL-7402) were treated with ABT-737 at various concentrations (5, 10, and 20 µM) for 24 and 48h. Cell morphology was examined under microscopy prior to each MTS assay to document cell death characteristics. Cell viability was assessed using MTS assay. BrdU incorporation assays were performed to specifically assess cell proliferation. Migration and invasion capabilities were evaluated through wound healing and transwell assays, respectively. To investigate PANoptosis pathway involvement, cells were co-treated with ABT-737 and specific inhibitors: Z-VAD-FMK (pan-caspase inhibitor, 10 µM), Necrostatin-1 (necroptosis inhibitor, 30 µM), or VX-765 (caspase-1 inhibitor, 10 µM). Flow cytometry analysis using Annexin V/PI staining was performed to directly assess cell death. Xenograft models were established in BALB/c nude mice using SK-HEP-1 and BEL-7402 cells to evaluate tumor formation with combination treatments. Tumor volumes were measured twice weekly, and tumor weights were recorded at the experimental endpoint. Ki-67 immunohistochemistry and H&E staining were performed on tumor sections to evaluate proliferation and necrosis. Western blot analysis was performed to examine the expression of PANoptosis-related proteins including phosphorylated MLKL (pMLKL) in both cultured cells and xenograft tumor tissues. ABT-737 demonstrated significant dose- and time-dependent inhibition of HCC cell proliferation. Microscopic examination revealed characteristic cell death morphology including cell shrinkage, membrane blebbing, and detachment in ABT-737-treated cells. Compared with the control group, ABT-737 treatment groups showed significantly reduced BrdU incorporation in a dose-dependent manner. Annexin V/PI flow cytometry demonstrated that compared with the control group, ABT-737 treatment groups exhibited significantly increased total apoptosis rates in a dose-dependent manner, confirming that the decrease in MTS absorbance reflects both reduced proliferation and increased cell death. Compared with the control group, ABT-737 treatment groups showed significantly reduced migration rates and invasive cell numbers. Co-treatment with PANoptosis pathway inhibitors partially restored cell viability compared with ABT-737 alone as measured by MTS assay. In xenograft models, compared with the control group, ABT-737 treatment group showed significantly reduced tumor volumes and tumor weights, which were partially reversed by pathway-specific inhibitors compared with ABT-737 alone. Compared with the control group, ABT-737-treated tumors showed significantly reduced Ki-67 positive rates, while H&E staining demonstrated markedly increased necrotic areas. Western blot analysis revealed that compared with the control group, ABT-737 treatment group showed upregulation of apoptosis markers (cleaved caspase-3, Bax), necroptosis markers (RIPK1, RIPK3, MLKL, and pMLKL), and pyroptosis markers (NLRP3, ASC, caspase-1), with concurrent downregulation of Bcl-2 in both cell lines and tumor tissues. ABT-737 exerts potent antitumor effects through the induction of PANoptosis in hepatocellular carcinoma, providing a promising therapeutic strategy for HCC treatment.

Neutrophil extracellular traps (NETs) constitute a vital antimicrobial defense mechanism of neutrophils, contributing to various physio-pathological processes; however, the role of plasma membrane asymmetry in this process remains unknown. Here we identify Xk-related protein 8 (XKR8), a plasma membrane phospholipid scramblase, as a pivotal regulator of NETs formation. Upon NETs induction, XKR8 is cleaved by caspase-3, thereby disrupting plasma membrane lipid asymmetry via phospholipid scrambling. Mutation of the caspase-3 cleavage site in XKR8 impairs NET formation. Inhibition of calcium signals before lipid scrambling abrogates NET formation, whereas blockade after scrambling does not. Cleaved XKR8 reorients plasma membrane lipids, altering membrane lipid tension and promoting Ca2+ signals through mechanosensitive channels. XKR8-deficient mice exhibit compromised NET formation and impaired control of Candida albicans pulmonary infection, showing that XKR8 is indispensable for neutrophil-driven immune responses in vivo. These findings define caspase-3-XKR8plasma membrane phospholipid scrambling as a central mechanism controlling NET formation and underscore its critical role in neutrophil-dependent antifungal immunity.

Self-association by small GTPases on membrane is critical for their signaling output and cellular function. However, a mechanistic understanding of how membrane components regulate this process remains incompletely understood. Here, we show that phosphatidylinositol 4,5-bisphosphate [PI(4,5)P2] promotes Arl4D self-association to potentiate downstream Pak1 signaling. We first show that Arl4D self-association is GTP-dependent and occurs at the plasma membrane. Fibronectin stimulation increases this self-association through two cooperative mechanisms: i) direct binding of PI(4,5)P2 by Arl4D via a conserved C-terminal polybasic motif, and ii) phosphorylation of Arl4D at Ser144 by its effector kinase Pak1. As a result, Arl4D membrane residency and protein stability are enhanced, with downstream signaling through Pak1 also amplified. Furthermore, pursuing structural prediction using AlphaFold, we generate an Arl4D mutant defective in self-association but retains GTP binding and membrane targeting, and find that this mutant fails to activate Pak1 for cell migration, while forced self-association of this mutant restores these downstream effects. Collectively, our findings reveal how an extracellular matrix cue leads to directional cell migration through Arl4D assembling into signaling-competent multimers at the plasma membrane, with cooperation between lipid recognition and kinase-mediated feedback playing critical roles.

OSCA (hyperosmolality-gated calcium-permeable channel) is an important class of mechanosensitive calcium channels in plants. OSCAs, which was located on the plasma membrane, function as osmosensors that enable plants to perceive hyperosmotic environments, such as drought and salt stress, and initiate downstream calcium ions signaling. However, a systematic genome-wide identification and functional characterization of the OSCA gene family in castor bean (Ricinus communis L.) under abiotic stresses remain unelucidated. In this study, we conducted a comprehensive genome-wide identification and analysis of the OSCA gene family in castor bean using bioinformatics approaches based on its whole-genome sequence. A total of 10 OSCA genes (RcOSCAs) were identified, which were unevenly distributed on five chromosomes. Phylogenetic analysis classified the RcOSCA proteins into four distinct subfamilies and clarified their evolutionary relationships with homologs from Arabidopsis thaliana, Oryza sativa, Triticum aestivum, Zea mays, and Glycine max. All proteins encoded by RcOSCA genes contain three conserved domains including RSN1_TM, PHM7_cyt, and RSN1_7TM. RT-qPCR analysis displayed that the transcript levels of several RcOSCA genes were significantly upregulated in roots, true leaves, or cotyledons under drought or salt stress. Notably, RcOSCA6 was rapidly and strongly induced in true leaves within 6h of salt stress, highlighting its potential key role in castor bean's adaptation to abiotic stress. This study constitutes the first genome-wide identification of the OSCA gene family in castor bean and provides preliminary insights into its evolutionary features and, more importantly, the species- and tissue-specific expression dynamics under salt and drought stresses. Future functional studies are warranted to fully elucidate the roles of these RcOSCAs in stress adaptation. These results contribute to understanding the molecular mechanisms underlying drought and salt tolerance in this resilient oil crop and offer valuable candidate genes for future functional characterization.

Anthracnose, caused by Colletotrichum siamense, is a major limiting factor for global natural rubber production. To develop sustainable control strategies, seven bacterial strains with antagonistic activity against C. siamense were isolated from healthy rubber tree leaves, with strain WR7 demonstrating the most significant antifungal effect, exhibiting an inhibition rate of 82.36%. Pot experiments revealed that WR7 achieved a disease control efficacy of 71.65% against C. siamense-induced anthracnose. Genomic analysis identified WR7 as Bacillus altitudinis. This strain inhibits pathogen growth through multiple mechanisms, including disruption of the pathogen’s cell wall and membrane integrity, induction of reactive oxygen species accumulation in hyphae, and secretion of cellulase, glucanase, protease, and siderophores. Gene cluster analysis further confirmed the potential of WR7 to synthesize antagonistic secondary metabolites such as lichenysin, fengycin, and bacilysin, while its sterile filtrate and volatile compounds also exhibited significant antifungal activity. Moreover, treatment with WR7 activated defense-related enzymes, including catalase and superoxide dismutase in rubber tree leaves, thereby enhancing the plant’s defense responses. This study is the first to report that Bacillus altitudinis WR7 has potential as a biocontrol agent for managing rubber tree anthracnose, offering a novel resource for sustainable disease management in rubber production.

Small molecule discovery and drug development are increasingly being pursued in academic settings, expanding beyond their traditional confinement to the pharmaceutical industry. The initial steps in drug discovery typically include identification and validation of a target, screening of chemical libraries to identify modulators of target activity, and subsequent prioritization and optimization of lead compounds using in vitro systems and animal models, with emphasis on compound potency, selectivity and pharmacological properties. This review focuses on early-stage discovery of small molecules that target plasma membrane transporters on epithelial cells, including absorptive and secretory epithelia in kidney, gastrointestinal tract, lung and eye. Of the estimated 500 distinct epithelial plasma membrane transporters, fewer than a dozen are the targets of approved drugs, most of which have been in clinical use for decades. We discuss the logistics and challenges associated with small molecule discovery in an academic setting. Specific epithelial cell targets are considered, including chloride channels, solute-coupled transporters, urea transporters and aquaporins, with therapeutic implications spanning constipation and secretory diarrheas, cystic fibrosis, dry eye disease, edema, hypertension and kidney stones. We conclude by identifying unmet needs and outlining opportunities to enable next-generation pharmacological modulation of epithelial transport processes.

Our previous studies have shown that major vault protein (MVP) is a virus-induced host factor that participates in the innate immune response. However, little is known about the role of MVP in Influenza A virus (IAV)- induced ferroptosis. In this study, the expression of MVP was found to positively correlate with that of interferon regulatory factor 1 (IRF1) and ferroptosis suppressor protein 1 (FSP1), but not with glutathione peroxidase 4 (GPX4), in peripheral blood mononuclear cells from patients with IAV. In vitro and in vivo evidence indicate that MVP is a potent factor in ferroptosis resistance during IAV infection. Upon investigating the mechanisms underlying this event, MVP was found to sequester IRF1 from tumor necrosis factor receptor-associated factor 6 (TRAF6), thereby suppressing its polyubiquitination and nuclear localization. Therefore, the transcription inhibition of IRF1 on the FSP1 promoter was removed, thereby enhancing FSP1 expression. A second wave of MVP regulation for IAV-induced ferroptosis also occurs. In the presence of the MVP, transcriptionally induced FSP1 is released from IRF1, leading to its ubiquitination and myristoylation, which enable its recruitment to the plasma membrane, where it functions as an oxidoreductase. These findings define a ferroptosis suppression pathway during IAV infection.

Drought is a major problem to mungbean ( Vigna radiata L.) productivity, necessitating the identification of tolerant genotypes and the exploration of their adaptive mechanisms. This study evaluated seven mungbean genotypes ‘BARI Mung-8’, ‘BMX-010015’, ‘K851’, ‘L-92’, ‘BARI Mung-1’, ‘FH-18’, and ‘PDM-139’ under control and drought treatments to characterize their physiological, biochemical, and molecular responses. Physiological traits, including chlorophyll content, photosynthesis rate (Pn), cell membrane stability (CMS), and relative water content (RWC), varied significantly (p≤ 0.05). Under drought, ‘BARI Mung-8’, ‘BMX-010015’, and ‘K851’ maintained chl content of 1.85–2.10 mg g -1 FW and Pn of 138–145 μmol m -2 s -1 , compared to 1.25 mg g -1 FW and 78 μmol m -2 s -1 in ‘BARI Mung-1’. These tolerant lines also retained high RWC (89–92%) and CMS (84–86%). Biochemically, they accumulated greater osmolytes, proline (38.7–42.1 µg g -1 FW) and glycine betaine (118–132 µg g -1 FW), and depicted enhanced antioxidant enzyme activities, including SOD (39.8–41.2 U mg -1 protein) and CAT (14.5–15.2 U mg -1 protein). Principal component analysis (PCA) and heatmap clustering grouped tolerant genotypes with these key adaptive traits, illustrating combined stress-response processes. Gene expression profiling showed significant upregulation (2.5–4.8 fold) of osmotic adjustment genes ( VrP5CS1 , VrBADH ), antioxidant defense genes ( VrSOD1 , VrCAT1 , VrPOD1 ), water transport gene ( VrPIP2-1 ), and stress signaling genes ( VrDREB2A , VrLEA3 ). The aquaporin gene VrPIP2–1 was associated with higher RWC, while VrCHLH stability supported chl retention. Integration of physiological, biochemical, and molecular data proved that drought tolerance in mungbean is regulated by coordinated cellular hydration, osmotic regulation, ROS detoxification, and transcriptional activation. “BARI Mung-8’, ‘BMX-010015’, ‘K851’, and ‘L-92’ emerged as eminent candidates for breeding programs targeting drought-prone environments, and the identified genes provide potential markers for selection of genotypes in climate-resilient legume improvement.

Pseudomonas syringae functions as a model phytopathogen causing numerous crop diseases, resulting in substantial economic losses in global agriculture. Presently, management of P. syringae predominantly depends on chemical pesticides; however, their prolonged application has contributed to escalating resistance and environmental contamination, highlighting urgent requirement for sustainable biological control approaches. In this review, we examine recent advances in the utilization and mechanistic understanding of natural products derived from plants, animals, and microorganisms for the control of P. syringae. Plant-derived compounds—including flavonoids, terpenoids, and alkaloids—inhibit P. syringae infection by targeting the bacterial type III secretion system (T3SS), disrupting cell membrane integrity, promoting reactive oxygen species (ROS) accumulation, and activating plant immune signaling pathways such as salicylic acid (SA) and jasmonic acid (JA) cascades. Animal-derived substances, such as chitosan, propolis, and antimicrobial peptides, primarily exert antibacterial effects through membrane disruption and immune system stimulation. Microbial-derived natural products contribute to synergistic disease suppression by modulating host immunity and interfering with the pathogen’s quorum sensing mechanisms. Evidence indicates that these natural products possess multi-target antimicrobial properties, offering a rich repository of candidate molecules, such as baicalein, lignans, and carvacrol, for the development of eco-friendly antibacterial agents. Future investigations should focus on detailed characterization of these bioactive compounds and their specific disease targets, optimization of extraction methodologies to improve stability and bioavailability, and comprehensive assessment of environmental safety to advance the industrial implementation of sustainable biocontrol strategies

Photodynamic therapy (PDT) faces significant challenges in clinical applications, including tumor hypoxia, the poor water solubility of photosensitizers, and insufficient targeting specificity. To address these limitations, we developed a biomimetic nanoagonist, 4T1@MFCB, by coloading a near-infrared photosensitizer (CyI) and a GLUT1 inhibitor (BAY-876) into a manganese/iron bimetallic metal-organic framework (MOF), followed by coating with the homologous 4T1 cell membrane. The manganese component catalyzes the decomposition of endogenous H2O2 to generate oxygen, alleviating tumor hypoxia and enhancing PDT efficacy, while the iron component promotes ferroptosis via the Fenton reaction. Meanwhile, BAY-876 inhibits glucose uptake, disrupting NADPH production and the cystine-to-cysteine reduction process, which leads to cystine accumulation and glutathione (GSH) depletion. These effects collectively suppress the Solute Carrier Family 7 Member 11(SLC7A11)/glutathione (GSH)/glutathione peroxidase 4 (GPX4) antioxidant axis, thereby triggering disulfidptosis. Under near-infrared irradiation, CyI mediates effective photodynamic and photothermal therapy (PTT). Both in vitro and in vivo studies demonstrate that 4T1@MFCB enables synergistic PDT/ferroptosis/disulfidptosis therapy, significantly inhibiting tumor growth without obvious systemic toxicity. This work highlights a multimodal treatment strategy that integrates metabolic intervention with nanocatalytic therapy, providing a promising approach for the precision treatment of hypoxic and therapy-resistant tumors.

The establishment and architectural growth of terrestrial plants critically depend on polar auxin transport, a process primarily driven by the asymmetric distribution of PIN proteins within the plasma membrane. In Arabidopsis, the exocyst, an evolutionarily conserved octameric vesicle-tethering complex, orchestrates the recycling of PIN proteins, with subunits such as SEC6, SEC8, and EXO70A1 having established roles in this process. However, how the exocyst is mechanistically integrated into PIN recycling networks and the regulatory hierarchy remain unsolved. Here, SEC3A driven by pollen-specific ProLAT52 promoter was transformed into sec3a/ + background, generating two independent pollen rescued (PRsec3a) mutant lines PRsec3a-1 and PRsec3a-2, and enabling investigation of SEC3A function in sporophytic growth related to PIN protein dynamics. PRsec3a exhibited stunted primary root elongation, disrupted gravitropism, and reduced auxin accumulation. Notably, endocytic recycling of PIN1, PIN2, and BRI1 proteins from the Brefeldin A-induced compartments was compromised in PRsec3a. Furthermore, SEC3A was identified as an effector of the small GTPase RabE1b, defining a novel regulatory axis. Together, these findings advance the understanding of Exocyst-mediated membrane trafficking and its pleiotropic roles in plant development.

Spontaneous hypertension (SH) is a prevalent chronic cardiovascular disorder characterized by the synergistic elevation of hydrogen peroxide (H2O2) levels and tyrosine hydroxylase (TH) activity in the brainstem nucleus tractus solitarius (NTS). Traditional detection techniques lack the specificity and spatiotemporal resolution to monitor the dynamic interplay of these two core pathological biomarkers, hindering the in-depth exploration of SH pathogenesis. Herein, a series of novel cascade-activated fluorescent probes (PPTHs) were rationally designed and synthesized based on a purine core, which achieve specific fluorescence responses only upon sequential activation by H2O2 and TH. In vitro assays demonstrated that the probes exhibited high sensitivity toward H2O2 in SH-SY5Y cell lysates, with a reliable limit of detection (LOD) and ideal anti-interference capability. Live-cell imaging further confirmed that purine-based molecules not only successfully mitigated probe adsorption on the cell membrane but also effectively improved the imaging signal-to-noise (S/N) ratio. Notably, PPTH-2-assisted confocal imaging clearly distinguished the differential fluorescence signals between normotensive control and SHR groups, which correlated with endogenous H2O2 level and TH activity in the NTS region. Our study presents a robust fluorescent probe platform for the synchronous detection of H2O2 and TH, offering a promising molecular tool for the early diagnosis and elucidation of the pathological mechanisms of SH.

Heterozygous pathogenic variants in ATP2B1 (encoding PMCA1) cause autosomal dominant intellectual developmental disorder 66 (MRD66; OMIM #619910). To date, only 12 pathogenic de novo ATP2B1 variants have been reported in MRD66. This study aimed to identify the genetic etiology in a Chinese infant with a neurodevelopmental disorder characterized by early-onset seizures and global developmental delay (GDD) and functionally characterize a novel ATP2B1 missense variant. Trio-based whole-exome sequencing revealed a heterozygous de novo ATP2B1 variant (c.2140A>C, p.Thr714Pro) in the proband. The proband presented with infantile spasms, GDD (Gesell Developmental Quotient: 65–74), and severe growth restriction (height/weight <−2 SD). To investigate the variant’s pathogenicity, the wild-type (WT) and mutant ATP2B1 constructs, N-terminally tagged with mScarlet, were transfected into HEK293T cells. Confocal imaging demonstrated profound cytoplasmic mislocalization of the p.Thr714Pro mutant protein, contrasting sharply with the characteristic plasma membrane localization of WT ATP2B1. Measurement of intracellular Ca 2+ levels using Fluo-4 AM showed a significant 2.07-fold increase in basal Ca 2+ levels in cells expressing the mutant compared to WT. This finding expands the spectrum of ATP2B1 variants associated with MRD66 and confirms calcium dyshomeostasis as the core pathomechanism. This case of MRD66 demonstrates a very early onset of seizures, consistent with the recognized phenotypic variability and the critical role of PMCA1 in early neurodevelopment.

Cyclic peptides are increasingly recognised as a therapeutic modality for modulating intracellular protein–protein interactions (PPIs), including those considered ‘undruggable’ by small molecules or biologics. Cyclic gomesin (cGm), an 18-residue β-hairpin peptide containing two disulfide bonds and a cyclised backbone, combines high chemical stability with amphipathic character that promotes selective interaction with negatively charged cancer cell membranes. We previously showed that cGm enters cancer cells at non-toxic concentrations by endocytosis and direct membrane partitioning, outperforming established cell-penetrating peptides, and that it can be engineered to incorporate a sequence that inhibits PPIs involved in lactate dehydrogenase-5 tetramerisation. Here, we further assess its grafting capacity by incorporating bioactive loop sequences of varying size, charge and hydrophobicity into the cGm framework. Structural and biophysical analyses confirmed that grafted analogues retained the three-dimensional fold and had membrane-binding features, anticancer activity, melanoma selectivity and low toxicity toward non-cancerous cells. These findings demonstrate the tolerance of cGm to sequence variation and support its development as a modular scaffold for designing intracellularly active cyclic peptide therapeutics.

Transcriptional control of lysosome biogenesis is an important mechanism underlying cellular adaptation to stress. It is largely unclear how cell surface changes or signals induce alteration in lysosome numbers. By developing a Caenorhabditis elegans-based heterologous TFE3 activation system, we here identify the non-receptor tyrosine kinases SRC-1/-2 (C. elegans) and FGR (mammals) as critical regulators of lysosome biogenesis. In C. elegans, inactivation of src-1/-2 leads to nuclear enrichment of ectopically expressed TFE3 and increased intensity of lysosomal markers. In mammalian cells, FGR inhibition or deficiency similarly results in TFEB/TFE3-dependent lysosomal increase. FGR acts through AKT2 by promoting the activation of the latter. FGR associates with the plasma membrane but is internalized onto endosomes and reaches lysosomes along the endosome-lysosome pathway following endocytosis. Lysosomal FGR promotes AKT2 recruitment to lysosomes, where it phosphorylates TFEB/TFE3 to prevent their activation. Together, these findings reveal a plasma membrane-to-lysosome signaling axis that is required for endocytosis-associated lysosome homeostasis.

In this study, gold nanoparticles were synthesized using an extract ofKalanchoe daigremontiana(K. daigremontiana) and biologically evaluated against the crude extract. The effects of both treatments were assessed in Jurkat T-cell leukemia cells, as a cancer model, and in 3T3-L1 fibroblasts, as a non-malignant control. Nanoparticle characterization revealed polyhedral AuNPs with an average diameter of 125.49 nm and an organic surface coating derived from the plant extract. Biological activity was evaluated using cell viability (MTT assay), intracellular reactive oxygen species (ROS) quantification, and plasma membrane permeability assays. TheK. daigremontiana-derived AuNPs exhibited selective, concentration-dependent cytotoxicity toward Jurkat cells through a multifactorial mechanism involving enhanced cellular uptake, oxidative stress induction, and membrane integrity disruption, while sparing non-malignant fibroblasts. In contrast, the crude extract induced a plateau-like cytotoxic response in 3T3-L1 fibroblasts (maximum cell death of 37.72%) and only mild cytotoxicity in Jurkat cells (18.23% at 150 μg ml-1), consistent with predominantly ROS-independent activity. This comparative analysis demonstrates that green synthesis of AuNPs fundamentally alters the biological mechanism ofK. daigremontiana, highlighting the added therapeutic potential of plant-derived AuNPs for leukemia treatment.

Red blood cells represent a widely used cellular model in cytotoxicity studies, particularly in hemocompatibility assessments. As enucleated cells, which are abundant and easily accessible in both humans and animals, red blood cells allow for rapid, reproducible, and low-cost evaluation of the toxicity of bioactive compounds, whether natural, synthetic, or nanoparticulate. From a functional perspective, the red blood cell membrane is highly sensitive to physical and chemical environmental changes (osmolarity, temperature, pH, and the presence of oxidizing agents). This sensitivity makes red blood cells an effective biosensor for detecting membrane damage, hemolysis, oxidative stress, methemoglobin formation, and aggregation processes. Therefore, in vitro tests using red blood cells allow for the preliminary evaluation in preclinical development, particularly for the early screening of cytotoxicity, membrane-disruptive effects, and hemocompatibility of small molecules, nanomaterials, and blood-contacting biomaterials. These techniques include hemocompatibility tests, evaluation of oxidative and osmotic damage, and evaluation of erythrocyte aggregation and function. However, the use of red blood cells as a cytotoxicity model also has significant limitations. As anucleate cells, erythrocytes lack organelles such as nuclei, mitochondria, or lysosomes, which prevents the evaluation of their effects on key intracellular processes such as protein synthesis, cell signaling, apoptosis, or endoplasmic reticulum stress. This lack of cellular complexity limits their usefulness as a sole model in studies of systemic toxicity or tissue-specific cytotoxicity. These tools offer an effective preliminary approach to anticipating risks in biomedical and pharmacological research.

Phase-separated plasma membranes show elevated viscosity and reduced line tension of liquid-ordered domains.

Repair of amyloid-β-induced plasma membrane damage via coordinated P21-activated kinase activation and Rab3a-directed vesicle fusion.