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ABSTRACTStreptococcus pneumoniae(S. pneumoniae) represents a major human bacterial pathogen leading to high morbidity and mortality in children and the elderly. Recent research emphasizes the role of extracellular vesicles (EVs) in bacterial pathogenicity. However, the contribution of S. pneumoniaeEVs (pEVs) to host-microbe interactions has remained unclear. Here, we observed that S. pneumoniaeinfections in mice led to severe lung injuries and alveolar epithelial barrier (AEB) dysfunction. Infections of S. pneumoniaereduced the protein expression of tight junction protein OCLN (occludin) and activated macroautophagy/autophagy in lung tissues of mice and A549 cells. Mechanically, S. pneumoniaeinduced autophagosomal degradation of OCLN leading to AEB impairment in the A549 monolayer. S. pneumoniaereleased the pEVs that could be internalized by alveolar epithelial cells. Through proteomics, we profiled the cargo proteins inside pEVs and found that these pEVs contained many virulence factors, among which we identified a eukaryotic-like serine-threonine kinase protein StkP. The internalized StkP could induce the phosphorylation of BECN1 (beclin 1) at Ser93 and Ser96 sites, initiating autophagy and resulting in autophagy-dependent OCLN degradation and AEB dysfunction. Finally, the deletion of stkPin S. pneumoniaecompletely protected infected mice from death, significantly alleviated OCLN degradation in vivo, and largely abolished the AEB disruption caused by pEVs in vitro. Overall, our results suggested that pEVs played a crucial role in the spread of S. pneumoniaevirulence factors. The cargo protein StkP in pEVs could communicate with host target proteins and even hijack the BECN1 autophagy initiation pathway, contributing to AEB disruption and bacterial pathogenicity.Abbreviations: AEB: alveolarepithelial barrier; AECs: alveolar epithelial cells; ATG16L1: autophagy related 16 like 1; ATP:adenosine 5’-triphosphate; BafA1: bafilomycin A1; BBB: blood-brain barrier; CFU: colony-forming unit; co-IP: co-immunoprecipitation; CQ:chloroquine; CTRL: control; DiO: 3,3′-dioctadecylox-acarbocyanineperchlorate; DOX: doxycycline; DTT: dithiothreitol; ECIS: electricalcell-substrate impedance sensing; eGFP: enhanced green fluorescentprotein; ermR: erythromycin-resistance expression cassette; Ery: erythromycin; eSTKs: eukaryotic-like serine-threoninekinases; EVs: extracellular vesicles; HA: hemagglutinin; H&E: hematoxylin and eosin; HsLC3B: human LC3B; hpi: hours post-infection; IP: immunoprecipitation; KD: knockdown; KO: knockout; LAMP1: lysosomal associated membrane protein 1; LC/MS: liquid chromatography-mass spectrometry; MAP1LC3/LC3: microtubule associated protein 1 light chain 3; MVs: membranevesicles; NC:negative control; NETs:neutrophil extracellular traps; OD: optical density; OMVs: outer membrane vesicles; PBS: phosphate-buffered saline; pEVs: S.pneumoniaeextracellular vesicles; protK: proteinase K; Rapa: rapamycin; RNAi: RNA interference; S.aureus: Staphylococcusaureus; SNF:supernatant fluid; sgRNA: single guide RNA; S.pneumoniae: Streptococcuspneumoniae; S.suis: Streptococcussuis; TEER: trans-epithelium electrical resistance; moi: multiplicity ofinfection; TEM:transmission electron microscope; TJproteins: tight junction proteins; TJP1/ZO-1: tight junction protein1; TSA: tryptic soy agar; WB: western blot; WT: wild-type

The eutectic medium-entropy alloys CoFeNiTax(x = 0.4, 0.5, 0.6) were designed successfully based on binary eutectic systems and thermodynamics calculations in this study. With the increase of Ta content, the solidified microstructures evolved from hypoeutectic to full eutectic and finally to hypereutectic. The primary phases in the hypoeutectic and hypereutectic were FCC solid solution and C14-Laves phase, respectively. The solidified microstructures were coarsened and spheroidized by the heat treatment, and a high-density of needle-like Laves phase precipitated in the FCC phase of the CoFeNiTa0.4. The crystallographic orientation relationship between the precipitated phase and matrix phase was [2‾42‾3]Laves//[011]FCC, and the crystal plane (1 1‾1)FCCwas almost parallel to (0 1‾12)Laves, just with a mismatch angle of about 6°. The compressive strength and fracture strain of the heat-treated alloys were within 2232∼2465 MPa and 22.5–32.8 %, respectively, being superior to most high-entropy alloys. The excellent combination of strength and toughness was attributed to the synergistic effects of various strengthening-toughening mechanisms, including the second-phase strengthening, the solid solution strengthening, the interface strengthening and toughening, and the twinning toughening.

When single-phase-to-ground (SPG) or phase-to-phase (PP) faults occur on collection lines of an offshore wind farm with modular multilevel converter-based high voltage direct current (MMC-HVDC) transmission, the fault current level may be low or even approach zero due to the controlled current at both ends of the collection line. As a result, the traditional current differential protection may malfunction. To address this issue, a method is proposed that utilizes open circuit characteristics of the negative-sequence network to indirectly calculate the fault point negative-sequence voltage by the negative-sequence voltage at the protection installation location. The fault distance can then be determined. The introduction of a twiddle factor eliminates the influence of fault resistance, improving the accuracy of fault distance calculation. Based on this calculated fault distance, a high-sensitivity differential protection scheme is proposed. This proposed protection can maintain sufficient sensitivity and trip correctly during collection line SPG and PP faults. It solves the issue of traditional current differential protection malfunctioning in low-fault current scenarios of transmission lines with inverter-based resources (IBRs) on both ends. In addition, it can adapt to different reactive power support strategies and operation modes of offshore wind farms. Test results indicate the effectiveness of the proposed protection.