Injury, stroke, and neurological diseases cause persistent accumulation of cellular debris that deteriorates lesion microenvironment and impedes central nervous system (CNS) repair. Debris clearance in the injured CNS has long been attributed primarily to microglia and infiltrating macrophages. Here, we identify perivascular cells as previously unrecognized phagocytes that expand after injury and exhibit robust phagocytic activity. Perivascular cell phagocytosis is conserved across multiple mouse models of CNS injury and human stroke lesions. These cells exhibit key hallmarks of phagocytosis, including LC3-associated phagocytosis for efficient lysosomal degradation. Mechanistically, phosphatidylserine serves as the eat-me signal and Axl mediates myelin debris uptake. Myelin phagocytosis drives perivascular cell proliferation, fibrosis and lesion progression. Genetic deletion of Axl in perivascular cells or pharmacological inhibition with the FDA-approved Axl inhibitor Gilteritinib reduces pathology and improves functional recovery after spinal cord injury. Together, these findings establish Axl-dependent perivascular cell phagocytosis as a therapeutic target for CNS repair.

Staphylococcus aureus is a facultative intracellular pathogen that persists within both professional and non-professional phagocytes, contributing to chronic and antibiotic-resistant infections. Autophagy, particularly xenophagy, serves as a central cell-autonomous defense pathway that can capture intracellular S. aureus and deliver it to lysosomes for degradation. However, the bacterium has evolved multiple strategies to subvert xenophagic clearance, including manipulation of bacteria-containing autophagosome maturation, blockade of autophagosome and lysosome fusion, and co-option of autophagy-related machinery to create intracellular survival niches. Recent studies have also identified host pathways that shape infection outcomes, including reprogramming of cell death cascades to simultaneously sustain host viability and suppress xenophagy, and co-option of mitophagy to eliminate bactericidal mitochondrial ROS. As illustrative examples, caspase-8 reprogramming uncouples host survival from effective xenophagy, and the HDAC11/IL10/mTOR/PINK1-PRKN axis drives mitophagy to suppress mitochondrial ROS. We further address the largely unexplored roles of chaperone-mediated autophagy and endosomal microautophagy in S. aureus infection, drawing on mechanistic paradigms from viral and mycobacterial infections. Since S. aureus can impair autophagosome maturation, lysosomal acidification, and fusion between bacteria-containing autophagosomes and lysosomes, enhancing autophagy initiation alone may not always translate into improved bacterial clearance. We therefore propose, as a testable hypothesis, that host-directed strategies aimed at restoring lysosomal competence, improving degradative flux, or neutralizing bacterial virulence mechanisms may complement or outperform upstream autophagy induction in selected infection contexts. However, this concept remains insufficiently validated, and direct comparative studies in relevant host cell types and animal models are needed before lysosome-directed approaches can be prioritized therapeutically.

Soybean aphid (Aphis glycines) is a major piercing-sucking pest of soybean, and drought frequently coincides with aphid outbreaks in the field. Whether exogenous inducers remain effective under water deficit, however, is poorly understood. We evaluated five inducers, including abscisic acid (ABA), β-aminobutyric acid (BABA), benzothiadiazole (BTH), methyl jasmonate (MeJA), and salicylic acid (SA), on soybean grown at 75%, 50%, and 25% field capacity (FC), and measured aphid population growth, defense enzyme activities, defense gene expression, reactive oxygen species (ROS), and callose deposition. All five inducers significantly suppressed aphid populations under every water regime, with MeJA providing the most stable protection. Defense responses formed a clear temporal cascade: defense genes were coordinately reprogrammed at 6 h after infestation (HAI), enzyme activities peaked at 120 HAI, and ROS bursts were closely coupled with callose deposition. Water status reshaped this cascade: ROS showed a rapid burst with efficient clearance under normal water, a strong burst with sustained maintenance under moderate drought, and a delayed burst with rapid collapse under severe drought. Partial least squares path modeling (PLS-PM) identified phenylalanine ammonia-lyase (PAL) as the only stable mediator across water regimes: the PAL2/PR1→PAL→Aphid pathway remained significant under 75% and 50% FC and explained 33.6%–39.3% of aphid variation, but collapsed under severe drought. Induced resistance under water deficit therefore rests largely on PAL-mediated defense under mild to moderate drought, and the effectiveness of any inducer depends on the water regime in which it is deployed.

Isoflavonoids, a major class of secondary metabolites predominantly found in legumes, serve as vital defenders against both biotic and abiotic stresses, though their molecular mechanisms in mitigating aluminum (Al) toxicity remain incompletely understood. In this study, we introduced the isoflavone synthase gene isolated from a white clover (TrIFS) into soybean. The resulting transgenic soybeans exhibited a noticeable increase in isoflavone accumulation in various organs and stronger tolerance to aluminum toxicity. The dry weight of roots of two transgenic lines, TL124 and TL129, were heavier than that of the wild type (WT) after 200 µM AlCl3 treatment, by 46.1 % and 38.6 % respectively. Higher seed yield per plant was also obtained in these two transgenic lines by 18.9 % and 18.2 % than in the WT under Al stress, respectively. Additionally, TrIFS transgenic soybeans showed significantly reduced levels of aluminum-induced malondialdehyde (MDA) and reactive oxygen species (ROS). RNA transcriptome analysis revealed that the transcriptional level of GmALDH22A2 (Glyma.09g036000, Wm82.a2.v1) was sharply decreased in the TrIFS transgenic lines under Al treatment. The GmALDH22A2 protein interaction with a respiratory burst oxidase homologue (GmRBOHL, Glyma.10g152200, Wm82.a2.v1) was comfirmed by yeast two hybrid, fluorescence complementation and co-immunoprecipitation assays. Furthermore, this ALDH-RBOH interaction could cause increase in ROS production in tobacco, which suggested that the decreased transcription of GmALDH22A2 in the TrIFS transgenic soybeans would attenuate ROS generation. In summary, we provided new insights in improving Al tolerance through isoflavone enhancement.

Integral membrane protein 2B (ITM2B), a transmembrane protein, frequently undergoes cleavage. The physiological functions of ITM2B are primarily studied in the context of neurological disorders, but their roles in cancers are largely overlooked. Here, it is demonstrated that in renal cell carcinoma (RCC) cells, N-terminal truncation of ITM2B facilitates migrasome swelling through the recruitment of TSPAN4 and promotes migrasome formation. Moreover, ITM2B truncation acts as a carrier, sorting active caspase-7 into migrasomes for migracytosis. The active caspase-7-enriched migrasomes are then taken up by macrophages, leading to caspase-7-induced IL-6 secretion from macrophages, which eventually aggravates RCC growth through a feedback mechanism. Physiologically, hyperuricemia enhances ITM2B cleavage to aggravate RCC growth. Clinically, RCC tissues tend to produce ITM2B truncations compared with corresponding para-carcinoma tissues. Moreover, compared with the urine from normal volunteers, that from RCC patients contains higher levels of ITM2B truncation-enriched migrasomes. This study not only highlights novel functions of ITM2B truncation in migrasome formation and active caspase-7 migracytosis but also elucidates the role of hyperuricemia in RCC progression via regulation of the ITM2B truncation-migrasome axis.

It has been shown that in most cells, low glucose leads to activation of AMP-activated protein kinase (AMPK) via the lysosomal glucose-sensing pathway, where glycolytic aldolase acts as the glucose sensor. Here, we show that ALDOC (aldolase C), the predominant isozyme of aldolase in mouse and rat oligodendrocyte precursor cells (OPCs), is acetylated at lysine 14, making the lysosomal glucose-sensing AMPK pathway unable to operate. We find that the blockage of AMPK activation is required for the proper proliferation and differentiation of OPCs into mature oligodendrocytes for myelination during development and for remyelination in areas of demyelination where the local glucose levels are low. Therefore, the acetylation of aldolase acts as a checkpoint for AMPK activation in response to low glucose to ensure the proliferation and differentiation of OPCs for myelination, and remyelination of demyelinated neurons. Inhibition of AMPK activation under low glucose conditions in oligodendrocyte precursor cells is shown to be important for myelination during development and remyelination in neuronal disorders.

Axoneme assembly constitutes a pivotal process in male gametogenesis of Plasmodium. Plasmodium possesses a unique nuclear envelope-anchored basal body that templates axoneme assembly, distinct from the basal body that templates the axoneme of cilia or flagella to protrude from the cell surface. In the canonical basal body, the microtubule (MT) triplet extends and forms the axonemal MT doublet. However, this characteristic MT triplet has not been detected in Plasmodium. Indeed, the MT organization and the mechanism underlying the axonemal MT doublet assembly remain elusive in Plasmodium. Here we utilize high-resolution imaging methods including iterative ultrastructure expansion microscopy (iU-ExM) and cryo-electron tomography (cryo-ET) to resolve the native MT organization in the basal body of male gametes from the rodent malaria parasite P. yoelii. The parasite exhibits an MT singlet-to-doublet transition, distinct from the canonical MT triplet-to-doublet transition. Furthermore, we reveal that δ-Tubulin and ε-Tubulin are expressed in male gametocytes and regulate axoneme formation during male gametogenesis. δ-Tubulin is localized at the proximal end of the MT B-tubule and modulates B-tubule assembly of MT doublet. Our work provides the native architecture of MT singlet-to-doublet transition and reveals the key role of δ-Tubulin and ε-Tubulin in MT singlet-to-doublet transition in the basal body of Plasmodium.

The cyclic GMP-AMP synthase (cGAS)/stimulator of IFN genes (STING) pathway is intimately associated with antitumoral immunity; however, the direct involvement of this pathway in tumor cell demise remains elusive. Here, we identified a compound, dodecyl 6-hydroxy-2-naphthoate (DHN), that induces pyroptosis in melanoma cells by activating noncanonical cGAS/STING signaling. DHN targets mitochondrial protein cyclophilin D (CypD) to induce the release of mitochondrial DNA, leading to cGAS activation and cyclic GMP-AMP (cGAMP) generation. Meanwhile, DHN-caused intracellular acidification induces protein kinase R-like endoplasmic reticulum kinase (PERK) activation, which promotes STING phosphorylation and polymerization in the presence of cGAMP, thereby facilitating the aggregation of STING in the ER, which serves as a platform to recruit Fas-associated via death domain (FADD) and caspase-8, leading to caspase-8 activation and subsequent gasdermin E cleavage, which ultimately results in pyroptosis of tumor cells and tumor regression in mouse models. The occurrence of this noncanonical cGAS/STING pathway-associated pyroptosis is also observed when both cGAS is activated and intracellular pH declines. Collectively, our findings reveal a pathway that links noncanonical cGAS/STING signaling to gasdermin E-mediated pyroptosis, thereby offering valuable insights for tumor therapy.

Microcystis aeruginosa blooms have become a global environmental concern. In particular, microcystins (MC) produced by algal cells pose a great threat to aquatic ecosystem security and human health. Therefore, it is necessary to develop environmentally friendly biological measures to reduce bloom hazards. In this study, a heterotrophic flagellate, NY1, that could quickly eliminate M. aeruginosa and efficiently remove MC was isolated and identified as Poteriospumella lacustris. When the initial cell density ratio of P. lacustris to M. aeruginosa was 1:200, P. lacustris rapidly ingested M. aeruginosa, and the clearance rates were more than 90 % within 24 h. P. lacustris effectively removed 83.06 % of MC (more than 200 mu g/L) in 24 h and 93.88 % in 72 h. Although temperatures lower than 15 degrees C and acidic conditions (pH 6) slightly and temporarily inhibited the ingestion and MC removal activities of P. lacustris within 24 h of treatment, it rapidly recovered and ingested more than 90 % of M. aeruginosa and eliminated 97 % of MC within 72 h at various temperatures (15-35 degrees C) and different pH values (pH 6-10). The fluorescence emission-excitation matrix spectra analysis and acute toxicity test showed that P. lacustris treatment reduced dissolved organic matter (DOM) and eliminated the toxicity of M. aerugionsa culture to three representative species at different trophic levels (Photobacterium phosphoreum, Daphnia magna, and Ctenopharyngodon idellus), indicating the promising potential of P. lacustris in improving water safety. In addition to M. aeruginosa, P. lacustris ingested and digested Platymonas subcordiformis, Chlorella vulgaris, and Synechocystis sp. This study provides a natural and biological solution to global Microcystis blooms by using the grazing effect of P. lacustris to remove toxic cyanobacteria and eliminate MC and DOM, ultimately improving water quality and safety.

Mutations in a Plasmodium de-ubiquitinase UBP1 have been linked to antimalarial drug resistance. However, the UBP1-mediated drug-resistant mechanism remains unknown. Through drug selection, genetic mapping, allelic exchange, and functional characterization, here we show that simultaneous mutations of two amino acids (I1560N and P2874T) in the Plasmodium yoelii UBP1 can mediate high-level resistance to mefloquine, lumefantrine, and piperaquine. Mechanistically, the double mutations are shown to impair UBP1 cytoplasmic aggregation and de-ubiquitinating activity, leading to increased ubiquitination levels and altered protein localization, from the parasite digestive vacuole to the plasma membrane, of the P. yoelii multidrug resistance transporter 1 (MDR1). The MDR1 on the plasma membrane enhances the efflux of substrates/drugs out of the parasite cytoplasm to confer multidrug resistance, which can be reversed by inhibition of MDR1 transport. This study reveals a previously unknown drug-resistant mechanism mediated by UBP1 through altered MDR1 localization and substrate transport direction in a mouse model, providing a new malaria treatment strategy.

The shift of carbon utilization from primarily glucose to other nutrients is a fundamental metabolic adaptation to cope with decreased blood glucose levels and the consequent decline in glucose oxidation. AMP-activated protein kinase (AMPK) plays crucial roles in this metabolic adaptation. However, the underlying mechanism is not fully understood. Here, we show that PDZ domain containing 8 (PDZD8), which we identify as a new substrate of AMPK activated in low glucose, is required for the low glucose-promoted glutaminolysis. AMPK phosphorylates PDZD8 at threonine 527 (T527) and promotes the interaction of PDZD8 with and activation of glutaminase 1 (GLS1), a rate-limiting enzyme of glutaminolysis. In vivo, the AMPK-PDZD8-GLS1 axis is required for the enhancement of glutaminolysis as tested in the skeletal muscle tissues, which occurs earlier than the increase in fatty acid utilization during fasting. The enhanced glutaminolysis is also observed in macrophages in low glucose or under acute lipopolysaccharide (LPS) treatment. Consistent with a requirement of heightened glutaminolysis, the PDZD8-T527A mutation dampens the secretion of pro-inflammatory cytokines in macrophages in mice treated with LPS. Together, we have revealed an AMPK-PDZD8-GLS1 axis that promotes glutaminolysis ahead of increased fatty acid utilization under glucose shortage.

Clinical information of patients with breast cancer.

The shift of carbon utilisation from glucose to other nutrients is a fundamental metabolic adaptation to cope with the decreased glucose oxidation during fasting or starvation 1 . AMP-activated protein kinase (AMPK) plays crucial roles in manifesting physiological benefits accompanying glucose starvation or calorie restriction 2 . However, the underlying mechanisms are unclear. Here, we show that low glucose-induced activation of AMPK plays a decisive role in the shift of carbon utilisation from glucose to glutamine. We demonstrate that endoplasmic reticulum (ER)-localised PDZD8, which we identify to be a new substrate of AMPK, is required for the glucose starvation-promoted glutaminolysis. AMPK phosphorylates PDZD8 at threonine 527 (T527), and promotes it to interact with and activate the mitochondrial glutaminase 1 (GLS1), a rate-limiting enzyme of glutaminolysis 3–5 , and as a result the ER-mitochondria contact is strengthened. In vivo, PDZD8 enhances glutaminolysis, and triggers mitohormesis that is required for extension of lifespan and healthspan in Caenorhabditis elegans subjected to glucose starvation or caloric restriction. Muscle-specific re-introduction of wildtype PDZD8, but not the AMPK-unphosphorylable PDZD8-T527A mutant, to PDZD8 −/− mice is able to rescue the increase of glutaminolysis, and the rejuvenating effects of caloric restriction in aged mice, including grip strength and running capacity. Together, these findings reveal an AMPK-PDZD8-GLS1 axis that promotes glutaminolysis and executes the anti-ageing effects of calorie restriction by promoting inter-organelle crosstalk between ER and mitochondria.

Supplementary materials and methods and supplementary figure legends.

This file contains 7 supplementary figures supporting that Flightless-I blocks p62-mediated recognition of LC3 to impede selective autophagy and promote breast cancer progression.

Morphogenesis of many protozoans depends on a polarized establishment of cortical cytoskeleton containing the subpellicular microtubules (SPMTs), which are apically nucleated and anchored by the apical polar ring (APR). In malaria parasite Plasmodium, APR emerges in the host-invading stages, including the ookinete for mosquito infection. So far, the fine structure and molecular components of APR as well as the underlying mechanism of APR-mediated apical positioning of SPMTs are largely unknown. Here, we resolve an unprecedented APR structure composed of a top ring plus approximate 60 radiating spines. We report an APR-localizing and SPMT-binding protein APR2. APR2 disruption impairs ookinete morphogenesis and gliding motility, leading to Plasmodium transmission failure in mosquitoes. The APR2-deficient ookinetes display defective apical anchorage of APR and SPMT due to the impaired integrity of APR. Using protein proximity labeling, we obtain a Plasmodium ookinete APR proteome and validate ten undescribed APR proteins. Among them, APRp2 and APRp4 directly interact with APR2 and also mediate the apical anchorage of SPMTs. This study sheds light on the molecular basis of APR in the organization of Plasmodium ookinete SPMTs.

为了探究UV?B辐射对大豆异黄酮合成调控的分子机理,本研究以UV?B辐射处理大豆V1期幼苗,采用HPLC和定量PCR方法分别测定处理前后各部位异黄酮含量和基因的表达,并克隆出GmUVR8光受体基因.发现UV?B辐射处理大豆V1期幼苗8 h后,叶中总异黄酮含量提高1.3倍,大豆苷元和染料木素分别提高15.8和16.5倍.大豆根、茎和叶中CHS和IFS基因对UV?B反应的时间和强度存在明显差异,CHS11在处理2 h后表达量增加38.4倍,IFS1和IFS2基因的最高表达量分别比处理前增加4.7和18.3倍.克隆出了大豆中编码UVR8光受体的GmUVR8a、GmUVR8b和GmUVR8c基因,其氨基酸序列与拟南芥AtUVR8的同源度为74％;GmUVR8a、GmUVR8b、GmUVR8c基因在叶中表达存在显著差异,以GmUVR8b表达量最高.结果表明在大豆中UV?B辐射可能通过多种UVR8光受体调控苯基丙酸类途径关键酶基因的表达,进而影响异黄酮的合成.

Dysfunction of protein trafficking has been intensively associated with neurological diseases, including neurodegeneration, but whether and how protein transport contributes to oligodendrocyte (OL) maturation and myelin repair in white matter injury remains unclear. ER-to-Golgi trafficking of newly synthesized proteins is mediated by coat protein complex II (COPII). Here, we demonstrate that the COPII component Sec13 was essential for OL differentiation and postnatal myelination. Ablation of Sec13 in the OL lineage prevented OPC differentiation and inhibited myelination and remyelination after demyelinating injury in the central nervous system (CNS), while improving protein trafficking by tauroursodeoxycholic acid (TUDCA) or ectopic expression of COPII components accelerated myelination. COPII components were upregulated in OL lineage cells after demyelinating injury. Loss of Sec13 altered the secretome of OLs and inhibited the secretion of pleiotrophin (PTN), which was found to function as an autocrine factor to promote OL differentiation and myelin repair. These data suggest that Sec13-dependent protein transport is essential for OL differentiation and that Sec13-mediated PTN autocrine signaling is required for proper myelination and remyelination.

Phaeocystis globosa causes severe marine pollution by forming harmful algal blooms and releasing hemolytic toxins and is therefore harmful to marine ecosystems and aquaculture industries. In this study, Microbulbifer sp. YX04 exerted high algicidal activity against P. globosa by producing and secreting metabolites. The algicidal activity of the YX04 supernatant was stable after exposure to different temperatures (-80 to 100°C) and pH values (4 to 12) for 2 h, suggesting that algicidal substances could temporarily be stored under these temperature and pH value conditions. To explore the algicidal process and mechanism, morphological and structural changes, oxidative stress, photosynthesis, autophagic flux, and global gene expression were investigated. Biochemical analyses showed that the YX04 supernatant induced reactive oxygen species (ROS) overproduction, which caused lipid peroxidation and malondialdehyde (MDA) accumulation in P. globosa. Transmission electron microscopy (TEM) observation and the significant decrease in both maximum photochemical quantum yield (Fv/Fm) and relative electron transfer rate (rETR) indicated damage to thylakoid membranes and destruction of photosynthetic system function. Immunofluorescence, immunoblot, and TEM analyses indicated that cellular damage caused autophagosome formation and triggered large-scale autophagic flux in P. globosa. Transcriptome analysis revealed many P. globosa genes that were differentially expressed in response to YX04 stress, most of which were involved in photosynthesis, respiration, cytoskeleton, microtubule, and autophagosome formation and fusion processes, which may trigger autophagic cell death. In addition to P. globosa, the YX04 supernatant showed high algicidal activity against Thalassiosira pseudonana, Thalassiosira weissflogii, Skeletonema costatum, Heterosigma akashiwo, and Prorocentrum donghaiense. This study highlights multiple mechanisms underlying YX04 supernatant toxicity toward P. globosa and its potential for controlling the occurrence of harmful algal blooms. IMPORTANCEP. globosa is one of the most notorious harmful algal bloom (HAB)-causing species, which can secrete hemolytic toxins, frequently cause serious ecological pollution, and pose a health hazard to animals and humans. Hence, screening for bacteria with high algicidal activity against P. globosa and studies on the algicidal characteristics and mechanism will contribute to providing an ecofriendly microorganism-controlling agent for preventing the occurrence of algal blooms and reducing the harm of algal blooms to the environment. Our study first reported the algicidal characteristic and mechanism of Microbulbifer sp. YX04 against P. globosa and demonstrated that P. globosa shows different response mechanisms, including movement ability, antioxidative systems, photosynthetic systems, gene expression, and cell death mode, to adapt to the adverse environment when algicidal compounds are present.

GLS1 orchestrates glutaminolysis and promotes cell proliferation when glutamine is abundant by regenerating TCA cycle intermediates and supporting redox homeostasis. CB-839, an inhibitor of GLS1, is currently under clinical investigation for a variety of cancer types. Here, we show that GLS1 facilitates apoptosis when glutamine is deprived. Mechanistically, the absence of exogenous glutamine sufficiently reduces glutamate levels to convert dimeric GLS1 to a self-assembled, extremely low-K-m filamentous polymer. GLS1 filaments possess an enhanced catalytic activity, which further depletes intracellular glutamine. Functionally, filamentous GLS1-dependent glutamine scarcity leads to inadequate synthesis of asparagine and mitogenome-encoded proteins, resulting in ROS-induced apoptosis that can be rescued by asparagine supplementation. Physiologically, we observed GLS1 filaments in solid tumors and validated the tumor-suppressive role of constitutively active, filamentous GLS1 mutants K320A and S482C in xenograft models. Our results change our understanding of GLS1 in cancer metabolism and suggest the therapeutic potential of promoting GLS1 filament formation.