USP18 Antagonizes Pyroptosis by Facilitating Selective Autophagic Degradation of Gasdermin D.

As a key executioner of pyroptosis, Gasdermin D (GSDMD) plays a crucial role in host defense and emerges as an essential therapeutic target in the treatment of inflammatory diseases. So far, the understanding of the mechanisms that regulate the protein level of GSDMD to prevent detrimental effects and maintain homeostasis is currently limited. Here, we unveil that ubiquitin-specific peptidase 18 (USP18) works as a negative regulator of pyroptosis by targeting GSDMD for degradation and preventing excessive innate immune responses. Mechanically, USP18 recruits E3 ubiquitin ligase mind bomb homolog 2 (MIB2) to catalyze ubiquitination on GSDMD at lysine (K) 168, which acts as a recognition signal for the selective autophagic degradation of GSDMD. We further confirm the alleviating effect of USP18 on LPS-triggered inflammation in vivo. Collectively, our study demonstrates the role of USP18 in regulating GSDMD-mediated pyroptosis and reveals a previously unknown mechanism by which GSDMD protein level is rigorously controlled by selective autophagy.

Open and closed microfluidics for biosensing.

Biosensing is vital for many areas like disease diagnosis, infectious disease prevention, and point-of-care monitoring. Microfluidics has been evidenced to be a powerful tool for biosensing via integrating biological detection processes into a palm-size chip. Based on the chip structure, microfluidics has two subdivision types: open microfluidics and closed microfluidics, whose operation methods would be diverse. In this review, we summarize fundamentals, liquid control methods, and applications of open and closed microfluidics separately, point out the bottlenecks, and propose potential directions of microfluidics-based biosensing.

Microfiber-Patterned Versatile Perfusable Vascular Networks.

Rapid construction of versatile perfusable vascular networks in vitro with cylindrical channels still remains challenging. Here, a microfiber-patterned method is developed to precisely fabricate versatile well-controlled perfusable vascular networks with cylindrical channels. This method uses tensile microfibers as an easy-removable template to rapidly generate cylindrical-channel chips with one-dimensional, two-dimensional, three-dimensional and multilayered structures, enabling the independent and precise control over the vascular geometry. These perfusable and cytocompatible chips have great potential to mimic vascular networks. The inner surfaces of a three-dimensional vascular network are lined with the human umbilical vein endothelial cells (HUVECs) to imitate the endothelialization of a human blood vessel. The results show that HUVECs attach well on the inner surface of channels and form endothelial tubular lumens with great cell viability. The simple, rapid and low-cost technique for versatile perfusable vascular networks offers plenty of promising opportunities for microfluidics, tissue engineering, clinical medicine and drug development.

Magnetic Janus origami robot for cross-scale droplet omni-manipulation.

The versatile manipulation of cross-scale droplets is essential in many fields. Magnetic excitation is widely used for droplet manipulation due to its distinguishing merits. However, facile magnetic actuation strategies are still lacked to realize versatile multiscale droplet manipulation. Here, a type of magnetically actuated Janus origami robot is readily fabricated for versatile cross-scale droplet manipulation including three-dimensional transport, merging, splitting, dispensing and release of daughter droplets, stirring and remote heating. The robot allows untethered droplet manipulation from ~3.2 nL to ~51.14 µL. It enables splitting of droplet, precise dispensing (minimum of ~3.2 nL) and release (minimum of ~30.2 nL) of daughter droplets. The combination of magnetically controlled rotation and photothermal properties further endows the robot with the ability to stir and heat droplets remotely. Finally, the application of the robot in polymerase chain reaction (PCR) is explored. The extraction and purification of nucleic acids can be successfully achieved.

Ferroptosis of Endothelial Cells Triggered by Erythrophagocytosis Contributes to Thrombogenesis in Uremia.

BACKGROUND: Although thrombosis events are the leading complication of uremia, their mechanism is largely unknown. The interaction between endothelial cells (ECs) and red blood cells (RBCs) in uremic solutes and its prothrombotic role need to be investigated. METHODS AND RESULTS: Here, we established an in vitro co-incubation model of uremic RBC and EC as well as a uremic rat model induced by adenine. Using flow cytometry, confocal microscopy, and electron microscopy, we found increased erythrophagocytosis by EC accompanied by increased reactive oxygen species, lipid peroxidation, and impairment of mitochondria, indicating that ECs undergo ferroptosis. Further investigations showed increased proteins' expression of heme oxygenase-1 and ferritin and labile iron pool accumulation in EC, which could be suppressed by deferoxamine (DFO). The ferroptosis-negative regulators glutathione peroxidase 4 and SLC7A11 were decreased in our erythrophagocytosis model and could be enhanced by ferrostatin-1 or DFO. In vivo, we observed that vascular EC phagocytosed RBC and underwent ferroptosis in the kidney of the uremic rat, which could be inhibited by blocking the phagocytic pathway or inhibiting ferroptosis. Next, we found that the high tendency of thrombus formation was accompanied by erythrophagocytosis-induced ferroptosis in vitro and in vivo. Importantly, we further revealed that upregulated TMEM16F expression mediated phosphatidylserine externalization on ferroptotic EC, which contributed to a uremia-associated hypercoagulable state. CONCLUSION: Our results indicate that erythrophagocytosis-triggered ferroptosis followed by phosphatidylserine exposure of EC may play a key role in uremic thrombotic complications, which may be a promising target to prevent thrombogenesis of uremia.

S-palmitoylation regulates innate immune signaling pathways: molecular mechanisms and targeted therapies.

S-palmitoylation is a reversible posttranslational lipid modification that targets cysteine residues of proteins and plays critical roles in regulating the biological processes of substrate proteins. The innate immune system serves as the first line of defense against pathogenic invaders and participates in the maintenance of tissue homeostasis. Emerging studies have uncovered the functions of S-palmitoylation in modulating innate immune responses. In this review, we focus on the reversible palmitoylation of innate immune signaling proteins, with particular emphasis on its roles in the regulation of protein localization, protein stability, and protein-protein interactions. We also highlight the potential and challenge of developing therapies that target S-palmitoylation or de-palmitoylation for various diseases.

Strain-induced recognition of molecular and chirality in cholesteric liquid crystal droplets for distance and curvature sensing.

Chirality is universal in nature and in biological systems, and the chirality of cholesteric liquid crystals (Ch-LC) is both controllable and quantifiable. Herein, a strategy for precise chirality recognition in a nematic LC host within soft microscale confined droplets is reported. This approach facilitates applications in distance and curvature sensing as well as on-site characterization of the overall uniformity and bending movements of a flexible device. Due to interfacial parallel anchoring, monodisperse Ch-LC spherical microdroplets show radial spherical structure (RSS) rings with a central radical point-defect hedgehog core. Strain-induced droplet deformation destabilizes the RSS configuration and induces the recognition of chirality, creating "core-shell" structures with distinguishable sizes and colors. In practice, an optical sensor is achieved due to the rich palette of optically active structures that can be utilized for gap distance measuring and the monitoring of curvature bending. The properties reported here and the constructed device have great potential for applications in soft robotics, wearable sensors, and advanced optoelectronic devices.

High-Strength Magnetic Hydrogels with Photoweldability Made by Stepwise Assembly of Magnetic-Nanoparticle-Integrated Aramid Nanofiber Composites.

Hydrogels capable of transforming in response to a magnetic field hold great promise for applications in soft actuators and biomedical robots. However, achieving high mechanical strength and good manufacturability in magnetic hydrogels remains challenging. Here, inspired by natural load-bearing soft tissues, a class of composite magnetic hydrogels is developed with tissue-mimetic mechanical properties and photothermal welding/healing capability. In these hydrogels, a hybrid network involving aramid nanofibers, Fe3O4 nanoparticles, and poly(vinyl alcohol) is accomplished by a stepwise assembly of the functional components. The engineered interactions between nanoscale constituents enable facile materials processing and confer a combination of excellent mechanical properties, magnetism, water content, and porosity. Furthermore, the photothermal property of Fe3O4 nanoparticles organized around the nanofiber network allows near-infrared welding of the hydrogels, providing a versatile means to fabricate heterogeneous structures with custom designs. Complex modes of magnetic actuation are made possible with the manufactured heterogeneous hydrogel structures, suggesting opportunities for further applications in implantable soft robots, drug delivery systems, human-machine interactions, and other technologies.

Modulation of virus-induced neuroinflammation by the autophagy receptor SHISA9 in mice.

Microglia and astrocytes are subgroups of brain glia cells that support and protect neurons within the central nervous system (CNS). At early stages of viral infection in the CNS, they are predominant responding cells and lead to recruitment of peripheral immune cells for viral clearance. Inhibitor of nuclear factor κB kinase subunit epsilon (IKKi) is critical for type I interferon signalling and inflammation, which modulate heterogenic immune responses during CNS infection. Balanced autophagy is vital to maintain brain integrity, yet regulation of autophagy and immune activity within brain glia cells is poorly understood. Here we identify SHISA9 as an autophagy cargo receptor that mediates the autophagy-dependent degradation of IKKi during herpes simplex virus type 1 infection. IKKi is recognized by SHISA9 through unanchored K48-linked poly-ubiquitin chains and bridged to autophagosome membrane components GABARAPL1. Single-cell RNA sequencing analysis shows that SHISA9 has temporal characteristics while modulating both antiviral and inflammatory responses in microglia and astrocytes at different stages during viral infection. We found that Shisa9-/- mice are highly susceptible to herpes simplex virus encephalitis, have pathogenic astrocytes and display more severe neuroinflammation compared with wild-type mice. Taken together, our study unravels a critical role of selective autophagy by orchestrating immune heterogeneity of different CNS resident cells through the SHISA9-IKKi axis.

Palmitoylation promotes chaperone-mediated autophagic degradation of NLRP3 to modulate inflammation.

The critical intracellular pattern recognition receptor NLRP3 senses pathogenic organisms and endogenous danger signals via forming inflammasomes to orchestrate innate immune responses. Dysfunction of NLRP3 inflammasomes is implicated in several inflammatory disorders. Hence, it is important to uncover the mechanisms preventing sustained NLRP3 inflammasome activation. Recently, we revealed that ZDHHC12-mediated palmitoylation enhances NLRP3 degradation through the chaperone-mediated autophagy pathway, and identified gain-of-function variants of NLRP3 in autoinflammatory diseases, which induce excessive NLRP3 inflammasome activation through decreased NLRP3 palmitoylation level and impaired chaperone-mediated autophagic degradation of NLRP3.

Metabolic enzyme UAP1 mediates IRF3 pyrophosphorylation to facilitate innate immune response.

Post-translational modifications (PTMs) of proteins are crucial to guarantee the proper biological functions in immune responses. Although protein phosphorylation has been extensively studied, our current knowledge of protein pyrophosphorylation, which occurs based on phosphorylation, is very limited. Protein pyrophosphorylation is originally considered to be a non-enzymatic process, and its function in immune signaling is unknown. Here, we identify a metabolic enzyme, UDP-N-acetylglucosamine pyrophosphorylase 1 (UAP1), as a pyrophosphorylase for protein serine pyrophosphorylation, by catalyzing the pyrophosphorylation of interferon regulatory factor 3 (IRF3) at serine (Ser) 386 to promote robust type I interferon (IFN) responses. Uap1 deficiency significantly impairs the activation of both DNA- and RNA-viruse-induced type I IFN pathways, and the Uap1-deficient mice are highly susceptible to lethal viral infection. Our findings demonstrate the function of protein pyrophosphorylation in the regulation of antiviral responses and provide insights into the crosstalk between metabolism and innate immunity.

Palmitoylation prevents sustained inflammation by limiting NLRP3 inflammasome activation through chaperone-mediated autophagy.

As a key component of the inflammasome, NLRP3 is a critical intracellular danger sensor emerging as an important clinical target in inflammatory diseases. However, little is known about the mechanisms that determine the kinetics of NLRP3 inflammasome stability and activity to ensure effective and controllable inflammatory responses. Here, we show that S-palmitoylation acts as a brake to turn NLRP3 inflammasome off. zDHHC12 is identified as the S-acyltransferase for NLRP3 palmitoylation, which promotes its degradation through the chaperone-mediated autophagy pathway. Zdhhc12 deficiency in mice enhances inflammatory symptoms and lethality following alum-induced peritonitis and LPS-induced endotoxic shock. Notably, several disease-associated mutations in NLRP3 are associated with defective palmitoylation, resulting in overt NLRP3 inflammasome activation. Thus, our findings identify zDHHC12 as a repressor of NLRP3 inflammasome activation and uncover a previously unknown regulatory mechanism by which the inflammasome pathway is tightly controlled by the dynamic palmitoylation of NLRP3.

Photonic Crystal-Integrated Optoelectronic Devices with Naked-Eye Visualization and Digital Readout for High-Resolution Detection of Ultratrace Analytes.

The detection of ultratrace analytes is highly desirable for the non-invasive monitoring of human diseases. However, a major challenge is fast, naked-eye, high-resolution ultratrace detection. Herein, a rectangular 3D composite photonic crystal (PC)-based optoelectronic device is first designed that combines the sensitivity-enhancing effects of PCs and optoelectronic devices with fast and real-time digital monitoring. A crack-free, centimeter-scale, mechanically robust ellipsoidal composite PCs with sufficient hardness and modulus, even exceeding most plastics and aluminum alloys, are developed. The high mechanical strength of ellipsoidal composite PCs allows them to be hand-machined into rectangular geometries that can be conformally covered with the centimeter-scale flat light-detection area without interference from ambient light, easily integrating 3D composite PC-based optoelectronic devices. The PC-based device's signal-to-noise ratio increases dramatically from original 30-40 to ≈60-70 dB. Droplets of ultratrace analytes on the device are identified by fast digital readout within seconds, with detection limits down to 5 µL, enabling rapid identification of ultratrace glucose in artificial sweat and diabetes risk. The developed 3D PC-based sensor offers the advantages of small size, low cost, and high reliability, paving the way for wider implementation in other portable optoelectronic devices.

Lentinan stabilized bimetallic PdPt3 dendritic nanoparticles with enhanced oxidase-like property for L-cysteine detection.

The development of nanozymes with enhanced catalytic activity has been drawing great interest. Lentinan with special structure may be used to prepare bimetallic nanomaterials to enhance their catalytic activity. Herein, lentinan stabilized PdPt3 dendritic nanoparticles (PdPt3-LNT NDs) were prepared through reduction of Na2PdCl4 and K2PtCl4 with a molar ratio of 1:3 using lentinan as a biological template. PdPt3-LNT NDs had dendritic shape with size of 10.76 ± 1.82 nm. PdPt3-LNT NDs had the hydrodynamic size about 25.7 nm and the zeta potential between -1.4 mV and - 4.9 mV at different pH. Furthermore, PdPt3-LNT NDs catalyzed 3,3',5,5'-tetramethylbenzidine (TMB) to produce oxidized TMB, suggesting their oxidase-like property. The catalytic activity of PdPt3-LNT NDs was the highest when pH was 4 and the temperature was 40 °C. The catalytic mechanism was the generation of reactive oxygen species- from O2 catalyzed by PdPt3-LNT NDs. More importantly, L-cysteine detection method was set up based on the oxidase-like property of PdPt3-LNT NDs. This method had wide linear range for 0-200 µM and low detection limit for 3.099 µM. Taken together, PdPt3-LNT NDs have good potential applications in bio-related detection in the future.

TMEM16F mediated phosphatidylserine exposure and microparticle release on erythrocyte contribute to hypercoagulable state in hyperuricemia.

The link between hyperuricemia (HUA) and the risk of venous thromboembolism (VTE) has been well established. However, the mechanisms of thrombus generation and the effect of HUA on procoagulant activity (PCA) of erythrocytes remain unclear no matter in uremia or hyperuricemia. Here, phosphatidylserine (PS) exposure, microparticles (MPs) release, cytosolic Ca2+, TMEM16F expression, reactive oxygen species (ROS) and lipid peroxidation of erythrocyte were detected by flow cytometer. PCA was assessed by coagulation time, purified coagulation complex and fibrin production assays. The fibrin formation was observed by scanning electron microscopy (SEM). We found that PS exposure, MPs generation, TMEM16F expression and consequent PCA of erythrocyte in HUA patients significantly increased compared to those in healthy volunteers. Furthermore, high UA induced PS exposure, and MPs release of erythrocyte in concentration and time-dependent manners in vitro, which enhanced the PCA of erythrocyte and was inhibited by lactadherin, a PS inhibitor. Additionally, using SEM, we also observed compact fibrin clots with highly-branched networks and thin fibers supported by red blood cells (RBCs) and RBC-derived MPs (RMPs). Importantly, we demonstrated UA enhanced the production of ROS and lipid peroxidation and reduced the generation of glutathione (GSH) of erythrocyte, which enhanced TMEM16F activity and followed PS externalization and RMPs formation. Collectively, these results suggest that Ca2+-dependent TMEM16F activation may be responsible for UA-induced PS exposure and MPs release of RBC, which thereby contribute to the prothrombotic risk in HUA.

Magnetic Field-Assisted Fission of a Ferrofluid Droplet for Large-Scale Droplet Generation.

With the presence of an external magnetic field, a ferrofluid droplet exhibits a rich variety of interesting phenomena notably different from nonmagnetic droplets. Here, a ferrofluid droplet impacting on a liquid-repellent surface is systematically investigated using high-speed imaging. The pre- and post-impact, including the droplet stretching, maximum spreading diameter, and final impact modes, are shown to depend on the impact velocity and the magnitude of the external magnetic field. A scaling relation involving the Weber and magnetic Bond numbers is fitted to predict the maximum spreading diameter based on the magnetic field-induced effective surface tension. The impact outcome is also investigated and classified into three patterns depending on the occurrence of the rim interface instability and the fission phenomenon. Two types of fission (i.e., evenly and unevenly distributed sizes of the daughter droplets) are first identified, and the corresponding mechanism is revealed. Last, according to Rayleigh-Taylor instability, a semiempirical formula is proposed to estimate the number of the daughter droplets in the regime of evenly distributed size, which agrees well with the experimental data. The present study can provide more insight into large-scale droplet generation with monodispersive sizes.

Droplet Bouncing: Fundamentals, Regulations, and Applications.

Droplet impact is a ubiquitous phenomenon in nature, daily life, and industrial processes. It is thus crucial to tune the impact outcomes for various applications. As a special outcome of droplet impact, the bouncing of droplets keeps the form of the droplets after the impact and minimizes the energy loss during the impact, being beneficial in many applications. A unified understanding of droplet bouncing is in high demand for effective development of new techniques to serve applications. This review shows the fundamentals, regulations, and applications of millimeter-sized droplet bouncing on solid surfaces and same/miscible liquids (liquid pool and another droplet). Regulation methods and current applications are summarized, and potential directions are proposed.

Microfluidic Applications in Drug Development: Fabrication of Drug Carriers and Drug Toxicity Screening.

Microfluidic technology has been highly useful in nanovolume sample preparation, separation, synthesis, purification, detection and assay, which are advantageous in drug development. This review highlights the recent developments and trends in microfluidic applications in two areas of drug development. First, we focus on how microfluidics has been developed as a facile tool for the fabrication of drug carriers including microparticles and nanoparticles. Second, we discuss how microfluidic chips could be used as an independent platform or integrated with other technologies in drug toxicity screening. Challenges and future perspectives of microfluidic applications in drug development have also been provided considering the present technological limitations.

Three-dimensional interconnected ultrathin manganese dioxide nanosheets grown on carbon cloth combined with Ti3C2Tx MXene for high-capacity zinc-ion batteries.

Aqueous zinc-ion batteries (ZIBs) are receiving a continuously increasing attention for the flexible and wearable electronics, due to their non-toxicity, non-flammability, and low-cost features. The development of high-performance flexible cathodes is of great significance to the development of flexible ZIBs. In this work, the flexible electrode of three-dimensional (3D) interconnected ultrathin MnO2 nanosheets on carbon cloth (CC@MnO2) coated with Ti3C2Tx MXene (CC@MnO2@MXene) is prepared by electrodeposition and dipping methods, in which CC@MnO2 is put into MXene dispersion for impregnation treatment to make the CC@MnO2 fibers wrapped with MXene completely. The results show that the coating of MXene improves the conductivity of the composite, and the interface between MXene and MnO2 provides more active sites. Therefore, CC@MnO2@MXene-10 electrode as the cathode of zinc ion battery provides high charge storage performance (517.0 mAh g-1 at 0.1 A g-1), excellent cycling stability (80.6 mAh g-1 after 800 cycles at 1 A g-1) and excellent energy density (701.3 Wh kg-1 at 133.8 W kg-1). Finally, flexible quasi-solid battery based on CC@MnO2@MXene composite as cathode was assembled, and the flexible electrodes show potential for application.