Single-nucleus transcriptome unveils the role of ferroptosis in ischemic stroke.

Multiple cell death pathways are involved in neuronal death in ischemic stroke (IS). However, the role of different cell death pathways in different cell types has not been elucidated. By analyzing three single-nucleus RNA sequencing (snRNA-seq) data of IS, we first found that a variety of programmed cell death (PCD) -related genes were significantly changed in different cell types. Based on machine learning and virtual gene knockout, we found that ferroptosis related genes, ferritin heavy chain 1 (Fth1) and ferritin light chain (Ftl1), play a key role in IS. Ftl1 and Fth1 can promote microglia activation, as well as the production of inflammatory factors and chemokines. Cell communication analysis showed that activated microglia could enhance chemotactic peripheral leukocyte infiltration, such as macrophages and neutrophils, through Spp1-Cd44 and App-Cd74 signaling, thereby aggravating brain tissue damage. Furthermore, real-time quantitative polymerase chain reaction (RT-qPCR) showed that P2ry12 and Mef2c were significantly decreased in oxygen-glucose deprivation (OGD) group, while Ftl1, Fth1, Apoe, Ctsb, Cd44 and Cd74 were significantly increased in OGD group. Collectively, our findings suggested targeted therapy against microglia Ftl1 and Fth1 might improve the state of microglia, reduce the infiltration of peripheral immune cells and tissue inflammation, and then improve the ischemic brain injury in mouse.

Phosphatidic acid-regulated SOS2 controls sodium and potassium homeostasis in Arabidopsis under salt stress.

The maintenance of sodium/potassium (Na+ /K+ ) homeostasis in plant cells is essential for salt tolerance. Plants export excess Na+ out of cells mainly through the Salt Overly Sensitive (SOS) pathway, activated by a calcium signal; however, it is unknown whether other signals regulate the SOS pathway and how K+ uptake is regulated under salt stress. Phosphatidic acid (PA) is emerging as a lipid signaling molecule that modulates cellular processes in development and the response to stimuli. Here, we show that PA binds to the residue Lys57 in SOS2, a core member of the SOS pathway, under salt stress, promoting the activity and plasma membrane localization of SOS2, which activates the Na+ /H+ antiporter SOS1 to promote the Na+ efflux. In addition, we reveal that PA promotes the phosphorylation of SOS3-like calcium-binding protein 8 (SCaBP8) by SOS2 under salt stress, which attenuates the SCaBP8-mediated inhibition of Arabidopsis K+ transporter 1 (AKT1), an inward-rectifying K+ channel. These findings suggest that PA regulates the SOS pathway and AKT1 activity under salt stress, promoting Na+ efflux and K+ influx to maintain Na+ /K+ homeostasis.

ESP-B4 promotes nasal epithelial cell-derived extracellular vesicles containing miR-146a-5p to modulate Smad3/GATA-3 thus relieving allergic rhinitis: ESP-B4/miR-146a-5p in AR.

BACKGROUND: Though generally a mild affliction, allergic rhinitis (AR) is very common and causes considerable discomfort. Ephedra sinica polysaccharide is a candidate cost-effective therapy to relieve AR symptoms. PURPOSE: We explore the molecular mechanism of pure polysaccharide ESP-B4 action in AR. METHODS: RPMI2650 cells were treated with lipopolysaccharide to induce an in vitro sensitization model, and extracellular vesicles (EVs) were isolated. A rat model of AR was established using ovalbumin as the allergen and was treated with Ephedra sinica polysaccharide to observe changes in rhinitis symptoms, nasal mucosa histopathology and molecular pathology. ESP-B4-treated sensitized cells were adopted in vitro to verify effect of Ephedra sinica polysaccharide on miR-146a-5p expression in RPMI2650 cell-derived EVs and helper T cell differentiation. RESULTS: miR-146a-5p inhibited Smad3, impeded the Smad3/GATA-3 interaction, upregulated IFN-γ expression, and promoted CD4+T cell Th1 differentiation. Treatment with ESP-B4 relieved AR in rats, and elevated miR-146a-5p in the EVs from the nasal epithelial cells, apparently in relation to effects on helper T cell Th1/Th2 equilibrium. CONCLUSION: Overall, ESP-B4 can promote miR-146a-5p secretion, affect the Th1/Th2 balance of helper T cells, and relieve AR symptoms through Smad3/GATA-3 interaction, thus presenting a potential strategy for AR treatment.

Identification of Aly1 and Aly2 as Modulators of Cytoplasmic pH in Saccharomyces cerevisiae.

The regulation of intracellular pH in yeast (Saccharomyces cerevisiae) cells is critical for cell function and viability. In yeast, protons (H+) can be excreted from the cell by plasma membrane ATPase PMA1 and pumped into vacuoles by vacuolar H+-ATPase. Because PMA1 is critical to the survival of yeast cells, it is unknown whether other compensatory components are involved in pH homeostasis in the absence of PMA1. To elucidate how intracellular pH is regulated independently of PMA1, we employed a screening approach by exposing the yeast haploid deletion mutant library (ver 4.0) to the selective plant plasma membrane H+-ATPase inhibitor PS-1, which we previously reported. After repeated screenings and verification, we identified two proteins, Aly1 and Aly2, that play a role in the regulation of intracellular pH when PMA1 is deficient. Our research uncovers a new perspective on the regulation of intracellular pH related to PMA1 and also preliminarily reveals a role for Aly1 and Aly2 in the regulation of intracellular pH.

PKS5 Confers Cold Tolerance by Controlling Stomatal Movement and Regulating Cold-Responsive Genes in Arabidopsis.

Cold stress limits plant growth and development; however, the precise mechanisms underpinning plant acclimation to cold stress remain largely unknown. In this study, the Ser/Thr protein kinase SOS2-LIKE PROTEIN KINASE5 (PKS5) was shown to play a positive role in plant responses to cold stress. A PKS5 loss-of-function mutant (pks5-1) exhibited elevated sensitivity to cold stress, as well as a lower survival rate and increased ion leakage. Conversely, PKS5 gain-of-function mutants (pks5-3, pks5-4) were more tolerant to cold stress and exhibited higher survival rates and decreased ion leakage. Stomatal aperture analysis revealed that stomatal closure was slower during the first 25 min after cold exposure in pks5-1 compared to wild-type, whereas pks5-3 and pks5-4 displayed accelerated stomatal closure over the same time period. Further stomatal aperture analysis under an abscisic acid (ABA) treatment showed slower closure in pks5-1 and more rapid closure in pks5-3 and pks5-4. Finally, expression levels of cold-responsive genes were regulated by PKS5 under cold stress conditions, while cold stress and ABA treatment can regulate PKS5 expression. Taken together, these results suggest that PKS5 plays a positive role in short-term plant acclimation to cold stress by regulating stomatal aperture, possibly via ABA pathways, and in long-term acclimation by regulating cold-responsive genes.

Phosphatidic acid inhibits SCAB1-mediated F-actin bundling in Arabidopsis.

Stomatal closure-associated actin-binding protein 1 (SCAB1) regulates stomatal closure by mediating actin filament reorganization in Arabidopsis thaliana. Our previous study showed that phosphatidylinositol 3-phosphate (PI3P) binds to SCAB1 and inhibits its oligomerization, thereby inhibiting its activity on F-actin in guard cells during stomatal closure. In this study, we show that another phospholipid, phosphatidic acid (PA), also binds to SCAB1 and inhibits its actin-bundling activity but not its actin-binding activity. F-actin bundling was promoted in vivo by treating Col-0 seedlings with n-butanol, a suppressor of PA production, but this effect was absent in the scab1 mutant. These results indicate that the signaling molecule PA is involved in the modulation of SCAB1 activity in F-actin reorganization.

High specific surface area N-doped activated carbon from hydrothermal carbonization of shaddock peel for the removal of norfloxacin from aqueous solution.

A novel N-doped activated carbon (NAC) derived from shaddock peel was investigated to remove norfloxacin (NFX) from aqueous solution. The Box-Behnken central composite design (BBD) was used to optimize the preparation conditions of NAC. The specific surface area of NAC was 2,481.81 m2 g-1, which was obtained at 1,106 K activation temperature, 2.4 h residence time, and 2.3:1 mass ratio of KOH to hydrochar. Moreover, the equilibrium data were perfectly represented by Langmuir and Koble-Corrigan isotherms, and the adsorption process was precisely described by the pseudo-second-order kinetic model. Besides, the adsorption of NFX on NAC was mainly controlled by π-π electron-donor-acceptor (EDA) interaction, hydrophobic effect, hydrogen-bonding, electrostatic interaction and Lewis acid-base effect. The maximum monolayer adsorption capacity of NFX was 746.29 mg g-1 at 298 K, implying that NAC was a promising adsorbent for the removal of NFX from aqueous solution.

Sharing Reward Program Based on Face Consciousness in Social Media.

Purpose: In social marketing, sharing reward program (SRP) is a common way to improve the marketing effect. However, few studies have explored the impact of consumers' self-presentation and face consciousness on enterprise SRP. This study aims to explore the influence of these two factors on the optimal SRP. Methods: A Stackelberg game between enterprises, sharers and potential consumers is developed to study the impact of sharers' face consciousness on enterprise's SRP. In order to discuss the impact of face consciousness on SRP in detail, we introduced status identity of commodity information, sharer's self-presentation preference and commodity price as exogenous variables in the research. Results: The results have shown that when the face consciousness of sharers is high, enterprises are advised to adopt the strategy of low reward and low requirement. But when the face consciousness is low, it would be better for them adopt the strategy of high reward and high requirement. In addition, with the low face consciousness, the optimal SRP is also affected by the relationship between the price of goods and the number of WeChat friends of sharers. Conclusion: The results suggest that when enterprises make incentive policies, considering consumers' self-presentation preference and face consciousness, the profit level can be effectively improved.

Influence of Mineralogical Structure of Mold Flux Film on Heat Transfer in Mold during Continuous Casting of Peritectic Steel.

The mineralogical structure of flux films is a critical factor in controlling heat transfer in the mold and avoiding the longitudinal cracking of slabs during the continuous casting of peritectic steel. In this study, the layered structure, crystallization ratio, mineralogical species, and morphology features of flux films were characterized by optical microscopy, X-ray diffraction, and electron-probe microanalysis. Microstructural observation revealed that the normal flux films for peritectic steels present a multilayered structure and high crystallization ratio (60~90 vol%), mainly composed of well-developed crystalline akermanite and cuspidine. In contrast, the films with outstanding flux characteristics with abundant longitudinal cracks on the slab surface have a low crystallization ratio (<50 vol%) or vast crystallite content (>80 vol%). Furthermore, heat transfer analysis showed that the low crystallization ratio and the vast crystallite content of flux films worsen the heat transfer rate or uniformity in the mold, whereas the appropriate thickness and cuspidine content of flux films can improve the heat transfer performance. From the above results, it is concluded that using strong crystalline flux to obtain the ideal mineral phase structure of flux film is one of the important measures for reducing longitudinal cracks during continuous casting of peritectic steel slabs.

Salt sealing induced in situ N-doped porous carbon derived from wheat bran for the removal of doxycycline from aqueous solution.

In situ N-doped porous carbon (NPC) derived from wheat bran via a convenient salt sealing and air-assisted strategy was prepared for the removal of doxycycline (DOX) from aqueous solution. The NPC was precisely characterized by SEM, FTIR, XPS and BET analysis. Additionally, the experimental variables including contact time, adsorbent dosage of NPC and pH were optimized by using Box-Behnken design (BBD) under response surface methodology (RSM). The predicted adsorption capacity of DOX was found to be 291.14 mg g-1 under optimalizing experimental conditions of 196 min contact time, 0.2 g L-1 adsorbent dosage and pH 5.78. The adsorption experimental data fitted Langmuir, Koble-Corrigan and Redlich-Peterson models well, and the pseudo-second-order model perfectly described the DOX adsorption process onto NPC. Thermodynamic parameters of DOX adsorbed onto NPC indicated that the adsorption process was spontaneous and endothermic. Moreover, the adsorption of DOX on NPC was mostly controlled by electrostatic interaction, π-π electron-donator-acceptor (EDA) interaction, hydrogen-bonding and Lewis acid-base effect. Besides, the N element of NPC also played a role in capturing DOX. The maximum monolayer adsorption capacity of DOX was turn out to be 333.23 mg g-1 at 298 K, which suggested that the NPC could be a prospectively adsorbent for the removal of DOX from wastewater.

Optimized preparation of activated carbon from furfural residue using response surface methodology and its application for bisphenol S adsorption.

Furfural residue (FR), a solid waste, was applied as the precursor to prepare activated carbon by steam activation. The Box-Behnken design (BBD) approach-based response surface methodology (RSM) was utilized to optimize the preparation conditions to evaluate their effects on the performance of activated carbon from furfural residue (FRAC). The optimum preparation conditions of FRAC were found as follows: activation temperature of 922 °C, activation time of 62 min, and the mass ratio of char to H2O of 1:4.5, resulting in 1,501.84 mg/g of iodine adsorption capacity and 1,662.41 m2/g of specific surface area. The FRAC was characterized and then the adsorption performance of bisphenol S (BPS) on FRAC was investigated. Langmuir and Koble-Corrigan isotherm models were well fitted to the experimental data, and the adsorption kinetics process was perfectly described by the pseudo-second-order model. Thermodynamic parameters showed that the adsorption of BPS was a spontaneous exothermic process. Besides, the regeneration efficiency of FRAC was over 97% after five consecutive cycles. The maximum monolayer adsorption capacity of FRAC for BPS was 3.2848 mmol/g at 298 K, indicating that the FRAC was an excellent adsorbent for the removal of BPS from aqueous solutions.

Phospholipids in Salt Stress Response.

High salinity threatens crop production by harming plants and interfering with their development. Plant cells respond to salt stress in various ways, all of which involve multiple components such as proteins, peptides, lipids, sugars, and phytohormones. Phospholipids, important components of bio-membranes, are small amphoteric molecular compounds. These have attracted significant attention in recent years due to the regulatory effect they have on cellular activity. Over the past few decades, genetic and biochemical analyses have partly revealed that phospholipids regulate salt stress response by participating in salt stress signal transduction. In this review, we summarize the generation and metabolism of phospholipid phosphatidic acid (PA), phosphoinositides (PIs), phosphatidylserine (PS), phosphatidylcholine (PC), phosphatidylethanolamine (PE) and phosphatidylglycerol (PG), as well as the regulatory role each phospholipid plays in the salt stress response. We also discuss the possible regulatory role based on how they act during other cellular activities.

Dynamic changes of phosphatidylinositol and phosphatidylinositol 4-phosphate levels modulate H+-ATPase and Na+/H+ antiporter activities to maintain ion homeostasis in Arabidopsis under salt stress.

Plant metabolites are dynamically modified and distributed in response to environmental changes. However, it is poorly understood how metabolic change functions in plant stress responses. Maintaining ion homeostasis under salt stress requires coordinated activation of two types of central regulators: plasma membrane (PM) H+-ATPase and Na+/H+ antiporter. In this study, we used a bioassay-guided isolation approach to identify endogenous small molecules that affect PM H+-ATPase and Na+/H+ antiporter activities and identified phosphatidylinositol (PI), which inhibits PM H+-ATPase activity under non-stress conditions in Arabidopsis by directly binding to the C terminus of the PM H+-ATPase AHA2. Under salt stress, the phosphatidylinositol 4-phosphate-to-phosphatidylinositol (PI4P-to-PI) ratio increased, and PI4P bound and activated the PM Na+/H+ antiporter. PI prefers binding to the inactive form of PM H+-ATPase, while PI4P tends to bind to the active form of the Na+/H+ antiporter. Consistent with this, pis1 mutants, with reduced levels of PI, displayed increased PM H+-ATPase activity and salt stress tolerance, while the pi4kß1 mutant, with reduced levels of PI4P, displayed reduced PM Na+/H+ antiporter activity and salt stress tolerance. Collectively, our results reveal that the dynamic change between PI and PI4P in response to salt stress in Arabidopsis is crucial for maintaining ion homeostasis to protect plants from unfavorable environmental conditions.

Study on the differential proteomics of rat hippocampal mitochondria during deep hypothermic circulatory arrest.

BACKGROUND: This study aimed to investigate the effect of deep hypothermic circulatory arrest (DHCA) on rat hippocampal mitochondrial protein expression and its differential proteomics, and explore the potential mechanisms behind the effect. METHODS: We used internal jugular vein reflux and tail artery perfusion methods to establish the rat cardiopulmonary bypass (CPB) model. Rats were dissected to obtain the hippocampus, and the hippocampal mitochondria were purified. The mitochondrial morphology and the mitochondrial marker cytochrome C oxidase (COX) qualitatively examined via transmission electron microscopy and western-blot analysis, respectively. The qualified samples were subjected to isobaric tags for relative and absolute quantification (iTRAQ); we then established the CPB model again to obtain the rat hippocampus for cryoultramicrotomy, and used immunofluorescent double staining technique to qualitatively and semi-quantitatively verify two representative differentially expressed proteins. RESULTS: By searching the Mascot 2.2 database, 29 differentially expressed proteins were obtained with statistical significance, including 21 known proteins and 8 unknowns. The expression level of COX and monoacylglycerol lipase did not change significantly (P>0.05) during the hyperacute phase; however, their intracellular localizations were altered. CONCLUSIONS: DHCA induced the differential expression of 29 rat hippocampal mitochondrial proteins, some of which had altered intracellular localization. We speculated that the localized alteration of these proteins is one of the neuroprotection mechanisms that occurs during DHCA.

Prime editing efficiently generates W542L and S621I double mutations in two ALS genes in maize.

Prime editing is a novel and universal CRISPR/Cas-derived precision genome-editing technology that has been recently developed. However, low efficiency of prime editing has been shown in transgenic rice lines. We hypothesize that enhancing pegRNA expression could improve prime-editing efficiency. In this report, we describe two strategies for enhancing pegRNA expression. We construct a prime editing vector harboring two pegRNA variants for W542L and S621I double mutations in ZmALS1 and ZmALS2. Compared with previous reports in rice, we achieve much higher prime-editing efficiency in maize. Our results are inspiring and provide a direction for the optimization of plant prime editors.

Green synthesis of stable Fe,Cu oxide nanocomposites from loquat leaf extracts for removal of Norfloxacin and Ciprofloxacin.

Mass production of nanomaterials to remove pollutants from water still faces many challenges, mainly due to the complexity of the synthesis methods involved and the use of dangerous reagents. The green method of preparation of nanomaterials from plants can effectively solve these problems. Fe,Cu oxide nanocomposites (Fe-Cu-NCs) were synthesized by a green and single-step method using loquat leaf extracts, and were used as an adsorbent for removal of Norfloxacin (NOR) and Ciprofloxacin (CIP) from aqueous solution. The synthesized adsorbent showed excellent adsorption properties for NOR and CIP. The experimental equilibrium data fitted the Redlich-Peterson and Koble-Corrigan models well and the maximum adsorption capacities of Fe-Cu-NCs calculated by the Langmuir model for NOR and CIP were 1.182 mmol/g and 1.103 mmol/g, respectively, at 293 K. Additionally, the morphologies and properties of Fe-Cu-NCs were characterized by transmission electron microscopy (TEM), scanning electron microscopy X-ray energy-dispersive spectroscopy (SEM-EDS), X-ray photoelectron spectroscopy (XPS), X-ray diffraction (XRD) and Fourier transform infrared spectroscopy (FTIR) analysis and the adsorption mechanism of NOR and CIP onto Fe-Cu-NCs was discussed. Thermodynamic parameters revealed that the adsorption process was spontaneous and endothermic. This study indicated that Fe-Cu-NCs are a potential adsorbent and provide a simple and convenient strategy for the purification of antibiotics-laden wastewater.

An improved protein lipid overlay assay for studying lipid-protein interactions.

BACKGROUND: Lipids perform multiple functions in the cell, and lipid-protein interactions play a key role in metabolism. Although various techniques have been developed to study lipid-protein interactions, the interacting protein partners that bind to most lipids remain unknown. The protein lipid overlay (PLO) assay has revealed numerous lipid-protein interactions, but its application suffers from unresolved technical issues. RESULTS: Herein, we found that blocking proteins may interfere with interactions between lipids and their binding proteins if a separate blocking step is carried out before the incubation step in the PLO assay. To overcome this, we modified the PLO assay by combining an incubation step alongside the blocking step. Verification experiments included phosphatidylinositol-3-phosphate (PI3P) and its commercially available interacting protein G302, C18:1, C18:2, C18:3 and the Arabidopsis plasma membrane H+-ATPase (PM H+-ATPase) AHA2 C-terminus, phosphatidylglycerol (PG) and AtROP6, and phosphatidylserine (PS) and the AHA2 C-terminus. The lipid-protein binding signal in the classical PLO (CPLO) assay was weak and not reproducible, but the modified PLO (MPLO) assay displayed significantly improved sensitivity and reproducibility. CONCLUSIONS: This work identified a limitation of the CPLO assay, and both sensitivity and reproducibility were improved in the modified assay, which could prove to be more effective for investigating lipid-protein interactions.

Corncob-derived activated carbon for roxarsone removal from aqueous solution: isotherms, kinetics, and mechanism.

In this study, the adsorption of roxarsone (ROX) onto corncob-derived activated carbon (AC) was optimized using response surface methodology (RSM). Following this, the AC was comprehensively characterized by FT-IR, SEM, and EDS analysis. The results showed that the highest ROX adsorption efficiency of 304.34 mg/g was obtained at the contact time of 262 min, initial pH of 2.5, adsorbent dosage of 0.4 g/L, and initial concentration of 240 mg/L. Besides, it was found that the adsorption equilibrium data was fitted well to the Langmuir and Sips isotherm models. The thermodynamic parameters (e.g., ΔG, ΔH, and ΔS) revealed the spontaneous and exothermic nature of ROX adsorption. As indicated by pseudo second-order kinetics model, the adsorption of ROX onto AC could be achieved through the hydrogen bond, π-π adsorbate-adsorbent interaction, and electrostatic interaction between AC surface functional group and molecular species variations of ROX at different pH values. Overall, it can be concluded that corncob-derived AC is an alternative option for removing ROX from aqueous solution.

The effect of interface atomic structure on the deformation mechanisms of Ti2AlN/TiAl composite under nanoindentation using MD simulations.

In this study, we conduct molecular dynamics simulations to investigate the effect of interface atomic structure on the deformation mechanisms of Ti2AlN/TiAl composite under nanoindentation. It is found that the interface atomic structure has a remarkable effect on the deformation behavior of Ti2AlN/TiAl composite due to significantly different interface-dislocation interactive mechanisms. For the Ti2AlN(0 0 0 1)/TiAl(1 1 1) coherent interface system, although plenty of dislocations nucleate and propagate through the TiAl layer during the indentation process, there are no dislocations transmitting across the coherent interface. The formation of stair-rod dislocation and dislocation tangle is the major mechanism for blocking the slip motion of dislocations by the coherent interface. Thus, the Ti2AlN(0 0 0 1)/TiAl(1 1 1) coherent interface is beneficial for the strengthening effect of Ti2AlN/TiAl composite. For the Ti2AlN(1 0 [Formula: see text] 3)/TiAl(1 1 1) incoherent interface system, the incoherent interface has some ability to annihilate the dislocations nucleated from the TiAl during the indentation process, but it cannot simultaneously annihilate plenty of dislocations. These dislocations accumulate in the incoherent interface and cause the stress concentration, providing the driving force for dislocation nucleation in Ti2AlN from the poor matching regions in the Al atomic arrays at the incoherent interface. Therefore, the incoherent interface can provide the access of dislocation transmission from TiAl to Ti2AlN, which benefits the ductility of Ti2AlN/TiAl composite.

FLOWERING LOCUS T2 regulates spike development and fertility in temperate cereals.

FLOWERING LOCUS T2 (FT2) is the closest paralog of the FT1 flowering gene in the temperate grasses. Here we show that overexpression of FT2 in Brachypodium distachyon and barley results in precocious flowering and reduced spikelet number, while down-regulation by RNA interference results in delayed flowering and a reduced percentage of filled florets. Similarly, truncation mutations of FT2 homeologs in tetraploid wheat delayed flowering (2-4 d) and reduced fertility. The wheat ft2 mutants also showed a significant increase in the number of spikelets per spike, with a longer spike development period potentially contributing to the delayed heading time. In the wheat leaves, FT2 was expressed later than FT1, suggesting a relatively smaller role for FT2 in the initiation of the reproductive phase. FT2 transcripts were detected in the shoot apical meristem and increased during early spike development. Transversal sections of the developing spike showed the highest FT2 transcript levels in the distal part, where new spikelets are formed. Our results suggest that, in wheat, FT2 plays an important role in spike development and fertility and a limited role in the timing of the transition between the vegetative and reproductive shoot apical meristem.