Calix[6]arene (CX6) was found to be an efficient ion transmembrane channel, which could be blocked by methylene blue (MB) through host-guest interactions. The blocked CX6 channel could be reopened by 4-sulfonated calix[6]arene owing to its stronger affinity with MB, thereby achieving a reversible ON-OFF-ON type switch.
Cells respond to the mechanical properties of the extracellular matrix (ECM) through formation of focal adhesions (FAs), re-organization of the actin cytoskeleton and adjustment of cell contractility. These are energy-demanding processes, but a potential causality between mechanical cues (matrix stiffness) and cellular (energy) metabolism remains largely unexplored. Here, we cultured human mesenchymal stem cells (hMSCs) on stiff (20 kPa) or soft (1 kPa) substrate and demonstrate that cytoskeletal reorganization and FA formation spreading on stiff substrates lead to a drop in intracellular ATP levels, correlating with activation of AMP-activated protein kinase (AMPK). The resulting increase in ATP levels further facilitates cell spreading and reinforces cell tension of the steady state, and coincides with nuclear localization of YAP/TAZ and Runx2. While on soft substrates (1 kPa), lowered ATP levels limit these cellular mechanoresponses. Furthermore, genetic ablation of AMPK lowered cellular ATP levels on stiff substrate and strongly reduced responses to substrate stiffness. Together, these findings reveal a hitherto unidentified relationship between energy expenditure and the cellular mechanoresponse, and point to AMPK as a key mediator of stem cell fate in response to ECM mechanics.
The determination of T4 polynucleotide kinase (PNK) activity and the screening of PNK inhibitors are critical to disease diagnosis and drug discovery. Numerous electrochemical strategies have been developed for the sensitive measurement of PNK activity and inhibition. However, they often suffer from additional labels and multiple steps of the detection process for the electrochemical readout. Herein, we have demonstrated an electrochemical DNA (E-DNA) sensor for the one-step detection of PNK with "signal-on" readout with no need for additional labels. In our design, the highly switchable double-stranded DNA (dsDNA) probes are immobilized on the gold nanoparticle-decorated molybdenum disulfide nanomaterial (MoS2-AuNPs), which possesses large surface area and high conductivity for elevating the signal gain in the PNK detection. This signal-on E-DNA sensor integrated with MoS2-AuNPs exhibits a much higher sensitivity than that without MoS2-AuNPs, showing a detection limit of 2.18 × 10–4 U/mL. Furthermore, this assay shows high selectivity, with the ability to discriminate PNK from other enzymes and proteins, and can be utilized to screen inhibitors. The proposed sensor is easy to operate with one-step readout and robust for PNK detection in the biological matrix and shows great potential for point-of-care in clinical diagnostics and drug screening.
Hyperactive angiogenesis contributes to the immunosuppressive microenvironment important for immunotherapy. Galectin‐1, encoded by LGALS1 , can trigger the vascular signaling programs and mediate the anti‐angiogenic treatment response. However, the mechanism through which galectin‐1 regulates angiogenesis is poorly understood. It has been suggested that galectin‐1 may associate with mRNAs in cells. This study applied the iRIP‐seq methodology to study the potential role of galectin‐1 as an RNA‐binding protein. We found that galectin‐1 interacts with a large number of mRNAs, with a preference for binding near stop codons and a preference for UGCA/UGGA and GAGCAG as binding motifs. Galectin‐1 binds to the mRNAs of angiogenesis‐associated genes including VEGFA , EGR1, and LAMA5 , suggesting that galectin‐1 may regulate angiogenesis via its mRNA‐binding activity. We further show that sh LGALS1 inhibits capillary tube formation in an in vitro angiogenesis assay and alters the expression levels of several galectin‐1‐bound angiogenesis‐associated mRNAs. These results uncover a previously unrecognized mRNA‐binding activity of galectin‐1.
In this study, a releasable disulfide carbonate linker is used to prepare disulfide-containing crosslinked polyethyleneimines (PEI-SS-CLs) for gene delivery. ESI-MS analysis shows that after being incubated with 1,4-dithio-DL-threitol (DTT), the degradable linkages in the polyethyleneimine (PEI) derivatives undergo disulfide bond cleavage followed by intramolecular cyclization and cleavage of the neighboring carbamate bond. Moreover, it is observed that thiol/polyanions trigger the release of DNA from polyplexes via the reductive degradability of PEI-SS-CLs and ion-exchange. In vitro transfection results indicate that PEI-SS-CL-1.5 (at a crosslinker/PEI feed molar ratio of 1.5) exhibits higher transfection efficacy than the commercially available reagents such as the high molecular weight PEI with a molecular weight of 25 kDa (PEI 25k) and the versatile liposomes Lipofectamine 2000. PEI-SS-CL-1.5 also demonstrates significantly lower cytotoxicity, compared with PEI 25k and Lipofectamine 2000. Our study indicates that incorporating the thiol-specific cleavable disulfide bond into crosslinked PEIs and implementing regulated release of DNA are effective strategies for designing safe and effective gene vectors.
As video generation models advance rapidly, assessing the quality of generated videos has become increasingly critical. Existing metrics, such as Fr\'echet Video Distance (FVD), Inception Score (IS), and ClipSim, measure quality primarily in latent space rather than from a human visual perspective, often overlooking key aspects like appearance and motion consistency to physical laws. In this paper, we propose a novel metric, VAMP (Visual Appearance and Motion Plausibility), that evaluates both the visual appearance and physical plausibility of generated videos. VAMP is composed of two main components: an appearance score, which assesses color, shape, and texture consistency across frames, and a motion score, which evaluates the realism of object movements. We validate VAMP through two experiments: corrupted video evaluation and generated video evaluation. In the corrupted video evaluation, we introduce various types of corruptions into real videos and measure the correlation between corruption severity and VAMP scores. In the generated video evaluation, we use state-of-the-art models to generate videos from carefully designed prompts and compare VAMP's performance to human evaluators' rankings. Our results demonstrate that VAMP effectively captures both visual fidelity and temporal consistency, offering a more comprehensive evaluation of video quality than traditional methods.
A perylene diimide-based metal-organic cage (MOC4c) was found to be an efficient transmembrane transporter for ions and small molecules through the internal cavity of the cage. MOC4c could selectively transport different anions, as evidenced by vesicle-based fluorescenceassays and planar lipid bilayer-based current recordings.Furthermore, MOC4c appears tofacilitate calcein transport across the lipid bilayer membrane of a livingcell, suggesting that MOC4c could be used as a biologicaltool for small molecule drugstransmembrane transportation.
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