Small extracellular vesicles (sEVs), which carry lipids, proteins and RNAs from their parent cells, serve as biomarkers for specific cell types and biological states. These vesicles, including exosomes and microvesicles, facilitate intercellular communication by transferring cellular components between cells. Current methods, such as ultracentrifugation and Tim‐4 affinity method, yield high‐purity sEVs. However, despite their small size, purified sEVs remain heterogeneous due to their varied intracellular origins. In this technical note, we used high‐speed atomic force microscopy (HS‐AFM) in conjunction with exosome markers (IgG ^CD63 and IgG ^CD81 ) to explore the intracellular origins of sEVs at single‐sEV resolution. Our results first revealed the nanotopology of HEK293T‐derived sEVs under physiological conditions. Larger sEVs (diameter > 100 nm) exhibited greater height fluctuations compared to smaller sEVs (diameter ≤ 100 nm). Next, we found that mouse‐origin IgG ^CD63 , and rabbit‐origin IgG ^control and IgG ^CD81 , exhibited the iconic ‘Y’ conformation, and similar structural dynamics properties. Last, exosome marker antibodies predominantly co‐localised with sEV _{d ≤ 100 nm} but not with sEV _{d > 100 nm} , demonstrating the CD63‐CD81‐enriched sEV and CD63‐CD81‐depleted sEV subpopulations. In summary, we demonstrate that nanoscopic profiling of surface exosome markers on sEVs using HS‐AFM is feasible for characterising distinct sEV subpopulations in a heterogeneous sEV mixture.

Nuclear pore complexes (NPCs) on the nuclear membrane surface have a crucial function in controlling the movement of small molecules and macromolecules between the cell nucleus and cytoplasm through their intricate core channel resembling a spiderweb with several layers. Currently, there are few methods available to accurately measure the dynamics of nuclear pores on the nuclear membranes at the nanoscale. The limitation of traditional optical imaging is due to diffraction, which prevents achieving the required resolution for observing a diverse array of organelles and proteins within cells. Super-resolution techniques have effectively addressed this constraint by enabling the observation of subcellular components on the nanoscale. Nevertheless, it is crucial to acknowledge that these methods often need the use of fixed samples. This also raises the question of how closely a static image represents the real intracellular dynamic system. High-speed atomic force microscopy (HS-AFM) is a unique technique used in the field of dynamic structural biology, enabling the study of individual molecules in motion close to their native states. Establishing a reliable and repeatable technique for imaging mammalian tissue at the nanoscale using HS-AFM remains challenging due to inadequate sample preparation. This study presents the rapid strainer microfiltration (RSM) protocol for directly preparing high-quality nuclei from the mouse brain. Subsequently, we promptly utilize HS-AFM real-time imaging and cinematography approaches to record the spatiotemporal of nuclear pore nano-dynamics from the mouse brain.

Background and Aim: For years, people have used sodium nitrite as a food preservative. This study determined the effect of okra (Abelmoschus esculentus L.) pod methanol extract (OPME) on mice with hepatotoxicity induced by sodium nitrite. The flavonoid and total phenolic levels, serum biochemistry, and liver histology were examined. Materials and Methods: Green okra pod extraction was performed using ethanol methanol solvent. Thirty adult male BALB/c mice (8-10 weeks, ∼30 g) were divided into six groups: Normal control, negative control (sodium nitrite 50 mg/kg BW exposure), and treatment groups (sodium nitrite exposure and OPME at doses of 50, 100, 200, and 400 mg/kg BW). Subsequently, they were exposed to sodium nitrite and administered multiple doses of OPME for 19 days by gavage. After that, serum was used for biochemical evaluation, and liver histological analysis was performed. All data were statistically analyzed (α=0.05). Results: All doses of OPME reduced the levels of nitric oxide (NO), malondialdehyde (MDA), alanine aminotransferase (ALT), and aspartate aminotransferase (AST). In this research, both superoxide dismutase (SOD) and catalase (CAT) levels increased in all OPME-administered treatments . All doses also reduced necrotic cells, proportion of swollen cells, and inflammation in liver histological analysis. The results of this study showed that OPME exerted hepatoprotective effects by lowering MDA, NO, ALT, and AST levels. It also improved SOD and CAT levels and recovered damaged liver tissue to its normal state. The optimal dose of OPME was 50-100 mg/kg BW. Conclusion: OPME has potential as a natural hepatoprotective agent against sodium nitrite exposure.

Cancer remains a major health issue, and Bcl-2 inhibits apoptosis. This study explored the phytochemicals of Codiaeum variegatum as potential Bcl-2 inhibitors. Receptor–ligand docking was conducted with YASARA, followed by 300 ns MD using AMBER14. ADME and DFT analyses were used to characterize the pharmacokinetics and electronic properties of the inhibitor candidates. The large-scale data generated from the 300-nanosecond MD simulations were further processed to map the energy landscapes using PCA and DCCM analysis in R Studio. We successfully applied the QM-AM1 method to the preparation and execution of molecular docking studies. Among the 17 phytochemical compounds isolated from C. variegatum, taraxerol demonstrated the most promising binding energy, with a docking score of −8.868 kcal/mol, which is comparable to that of the co-crystallized reference ligand, venetoclax. Taraxerol also satisfied the ADME criteria based on Lipinski’s Rule of Five, with only a single rule violation, and DFT calculations confirmed its stability, exhibiting an energy gap of −0.1805 eV. Furthermore, a 300 ns MD simulation revealed that the binding energy of taraxerol, computed using the MM-PBSA method, was more favorable than that of venetoclax, with taraxerol yielding 303.235 kJ/mol in complex with Bcl-2, compared to 291.115 kJ/mol for venetoclax. PCA and DCCM analyses additionally supported the dynamic stability of taraxerol as a potential Bcl-2 inhibitor. Although ADME predictions suggest a favorable safety profile, further in vitro and in vivo validation is required to substantiate its therapeutic potential.

Selenium nanoparticles (SeNPs) have gained attention in the medical field because of their enhanced bioactive properties and bioavailability. The green synthesis of nanoparticles offers an eco-friendly synthesis method. In this study, Abelmoschus esculentus (okra) extract was used for the green synthesis of SeNPs. The synthesized SeNPs were characterized using UV-visible (UV-vis), scanning electron microscopy-energy dispersive X-ray (SEM-EDX), Fourier-transform infrared spectroscopy (FTIR), and particle size analyzer (PSA). UV-vis analysis showed a peak at a wavelength of 288 nm, and the SeNPs demonstrated stability at low temperatures for up to 30 days. FTIR analysis indicated interactions between the pectin functional groups on okra and the surface of SeNPs. The SeNPs had a 6.48 nm diameter and polydispersity index of 0.13. A spherical morphology was observed in the nanoparticles. The antioxidant activity of SeNPs was evaluated using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid (ABTS) methods. The results showed a dose-dependent increase in free radical inhibition with 31.44 and 15.37% by DPPH and ABTS tests, respectively. The green synthesis of SeNPs using okra extract produced nanoparticles with unique properties based on several characterization results. In conclusion, SeNPs exhibited promising antioxidant activity, highlighting their potential for biomedical applications.

Microtubule catastrophe is widely attributed to an end-driven loss of the GTP cap, implying that structural failure is confined to filament termini. Here, we show that this view is incomplete. Using high-speed atomic force microscopy (HS-AFM), we directly visualize nanoscale structural failure in paclitaxel-treated microtubules in real time. We identify spontaneous lattice nanodefects that emerge within the filament shaft and act as internal initiation sites for disassembly. These defects arise from localized protofilament decoupling and lattice strain accumulation, which disrupt the cooperative interactions within the microtubule wall. Once a critical threshold is reached, these sites nucleate disassembly that propagates as distinct collapse trajectories, including bidirectional spreading from intralattice defects and inward convergence from opposing ends, in addition to classical end-initiated depolymerization. Rather than uniformly stabilizing the lattice, paclitaxel maintains microtubules in a metastable state in which internal stress heterogeneity persists and enables defect-driven failure. Our results establish lattice decoupling as a primary structural trigger of microtubule catastrophe and redefine it as a defect-mediated, spatially encoded transition. These findings provide a nanoscale mechanistic framework for understanding heterogeneous responses to microtubule-targeting chemotherapeutics, suggesting that drug resistance may arise from modulation of lattice integrity and defect propagation rather than drug binding alone.