Extracellular pH sensing by plant cell-surface peptide-receptor complexes.

The extracellular pH is a vital regulator of various biological processes in plants. However, how plants perceive extracellular pH remains obscure. Here, we report that plant cell-surface peptide-receptor complexes can function as extracellular pH sensors. We found that pattern-triggered immunity (PTI) dramatically alkalinizes the acidic extracellular pH in root apical meristem (RAM) region, which is essential for root meristem growth factor 1 (RGF1)-mediated RAM growth. The extracellular alkalinization progressively inhibits the acidic-dependent interaction between RGF1 and its receptors (RGFRs) through the pH sensor sulfotyrosine. Conversely, extracellular alkalinization promotes the alkaline-dependent binding of plant elicitor peptides (Peps) to its receptors (PEPRs) through the pH sensor Glu/Asp, thereby promoting immunity. A domain swap between RGFR and PEPR switches the pH dependency of RAM growth. Thus, our results reveal a mechanism of extracellular pH sensing by plant peptide-receptor complexes and provide insights into the extracellular pH-mediated regulation of growth and immunity in the RAM.

EIN2-directed translational regulation of ethylene signaling in Arabidopsis.

Ethylene is a gaseous phytohormone that plays vital roles in plant growth and development. Previous studies uncovered EIN2 as an essential signal transducer linking ethylene perception on ER to transcriptional regulation in the nucleus through a "cleave and shuttle" model. In this study, we report another mechanism of EIN2-mediated ethylene signaling, whereby EIN2 imposes the translational repression of EBF1 and EBF2 mRNA. We find that the EBF1/2 3' UTRs mediate EIN2-directed translational repression and identify multiple poly-uridylates (PolyU) motifs as functional cis elements of 3' UTRs. Furthermore, we demonstrate that ethylene induces EIN2 to associate with 3' UTRs and target EBF1/2 mRNA to cytoplasmic processing-body (P-body) through interacting with multiple P-body factors, including EIN5 and PABs. Our study illustrates translational regulation as a key step in ethylene signaling and presents mRNA 3' UTR functioning as a "signal transducer" to sense and relay cellular signaling in plants. VIDEO ABSTRACT.

NLRs guard metabolism to coordinate pattern- and effector-triggered immunity.

Pattern-triggered immunity (PTI) and effector-triggered immunity (ETI) in plants enable them to respond to pathogens by activating the production of defence metabolites that orchestrate immune responses1-4. How the production of defence metabolites is promoted by immune receptors and coordinated with broad-spectrum resistance remains elusive. Here we identify the deubiquitinase PICI1 as an immunity hub for PTI and ETI in rice (Oryza sativa). PICI1 deubiquitinates and stabilizes methionine synthetases to activate methionine-mediated immunity principally through biosynthesis of the phytohormone ethylene. PICI1 is targeted for degradation by blast fungal effectors, including AvrPi9, to dampen PTI. Nucleotide-binding domain, leucine-rich-repeat-containing receptors (NLRs) in the plant immune system, such as PigmR, protect PICI1 from effector-mediated degradation to reboot the methionine-ethylene cascade. Natural variation in the PICI1 gene contributes to divergence in basal blast resistance between the rice subspecies indica and japonica. Thus, NLRs govern an arms race with effectors, using a competitive mode that hinges on a critical defence metabolic pathway to synchronize PTI with ETI and ensure broad-spectrum resistance.

A molecular framework of ethylene-mediated fruit growth and ripening processes in tomato.

Although the role of ethylene in tomato (Solanum lycopersicum) fruit ripening has been intensively studied, its role in tomato fruit growth remains poorly understood. In addition, the relationship between ethylene and the developmental factors NON-RIPENING (NOR) and RIPENING INHIBITOR (RIN) during ripening is under debate. Here, we carried out comprehensive genetic analyses of genome-edited mutants of tomato ETHYLENE INSENSITIVE 2 (SlEIN2), four EIN3-like genes (SlEIL1-4), and three EIN3 BINDING F-box protein genes (SlEBF1-3). Both slein2-1 and the high-order sleil mutant (sleil1 sleil2 sleil3/SlEIL3 sleil4) showed reduced fruit size, mainly due to decreased auxin biosynthesis. During fruit maturation, slein2 mutants displayed the complete cessation of ripening, which was partially rescued by slebf1 but not slebf2 or slebf3. We also discovered that ethylene directly activates the expression of the developmental genes NOR, RIN, and FRUITFULL1 (FUL1) via SlEIL proteins. Indeed, overexpressing these genes partially rescued the ripening defects of slein2-1. Finally, the signal intensity of the ethylene burst during fruit maturation was intimately connected with the progression of full ripeness. Collectively, our work uncovers a critical role of ethylene in fruit growth and supports a molecular framework of ripening control in which the developmental factors NOR, RIN, and FUL1 act downstream of ethylene signaling.

Chemical genetic screening identifies nalacin as an inhibitor of GH3 amido synthetase for auxin conjugation.

Auxin inactivation is critical for plant growth and development. To develop plant growth regulators functioning in auxin inactivation pathway, we performed a phenotype-based chemical screen in Arabidopsis and identified a chemical, nalacin, that partially mimicked the effects of auxin. Genetic, pharmacological, and biochemical approaches demonstrated that nalacin exerts its auxin-like activities by inhibiting indole-3-acetic acid (IAA) conjugation that is mediated by Gretchen Hagen 3 (GH3) acyl acid amido synthetases. The crystal structure of Arabidopsis GH3.6 in complex with D4 (a derivative of nalacin) together with docking simulation analysis revealed the molecular basis of the inhibition of group II GH3 by nalacin. Sequence alignment analysis indicated broad bioactivities of nalacin and D4 as inhibitors of GH3s in vascular plants, which were confirmed, at least, in tomato and rice. In summary, our work identifies nalacin as a potent inhibitor of IAA conjugation mediated by group II GH3 that plays versatile roles in hormone-regulated plant development and has potential applications in both basic research and agriculture.

Modulating Aryl Azide Photolysis: Synthesis of a Room-Temperature Phosphorescent Carboline in Cucurbit[7]uril Host.

Cucurbit[7]uril (CB7), a supramolecular host, is employed to control the pathway of photolysis of an aryl azide in an aqueous medium. Normally, photolysis of aryl azides in bulk water culminates predominantly in the formation of azepine derivatives via intramolecular rearrangement. Remarkably, however, when this process unfolds within the protective confinement of the CB7 cavity, it results in a carboline derivative, as a consequence of a C─H amination reaction. The resulting carboline caged by CB7 reveals long-lived room temperature phosphorescence (RTP) in the solid state, with lifetimes extending up to 2.1 s. These findings underscore the potential of supramolecular hosts to modulate the photolysis of aryl azides and to facilitate novel phosphorescent materials.

Cathepsin V is correlated with the prognosis and tumor microenvironment in liver cancer.

Recent studies have shown that high cell cycle activity negatively correlates with antitumor immunity in certain cancer types. However, a similar correlation has not been proven in liver cancer. We downloaded transcriptomic profiles of the cancer genome atlas-liver hepatocellular carcinoma (TCGA-LIHC) and assessed the cell cycle distribution of samples using single sample gene set enrichment analysis (ssGSEA), termed the cell cycle score (CCS). We obtained cell cycle-related differentially expressed prognostic genes and identified CENPA, CDC20, and CTSV using LASSO regression. We studied the effect of CTSV on clinical features and immune alterations in liver cancer based on TCGA-LIHC data. In vitro and in vivo experiments were performed to validate the role of CTSV in liver cancer using liver cancer cell lines and tissues. We found that the CCS closely correlated with the clinical features and prognosis of patients in TCGA-LIHC. Analysis of differentially expressed genes (DEGs), univariate Cox regression, and least absolute shrinkage and selection operator (LASSO) regression identified cathepsin V (CTSV) with prognostic significance in LIHC. Importantly, single-gene survival analysis of CTSV using microarray and sequencing data indicated that high levels of CTSV expression correlated with an unfavorable prognosis in various cancers. Gene set enrichment analysis revealed that high CTSV expression closely correlated with decreased expression of metabolic genes and increased expression of cell cycle genes. Furthermore, difference and correlation analyses of the relationship between CTSV expression and immune infiltrates, determined using CIBERSORT and TIMER algorithms, revealed that CTSV expression correlated with macrophages and CD4+ T cells. In vitro and in vivo experiments revealed that knockdown of CTSV inhibited liver cancer cells proliferation. Immunohistochemical staining showed that high CTSV expression correlated with macrophage infiltration in liver cancer tissues, predicted a poor prognosis, and is associated with the effectiveness of hepatocellular carcinoma treatment. In couclusion, CTSV is a novel cell cycle-associated gene with clinical significance in HCC.

EIN3 and RSL4 interfere with an MYB-bHLH-WD40 complex to mediate ethylene-induced ectopic root hair formation in Arabidopsis.

The alternating cell specifications of root epidermis to form hair cells or nonhair cells in Arabidopsis are determined by the expression level of GL2, which is activated by an MYB-bHLH-WD40 (WER-GL3-TTG1) transcriptional complex. The phytohormone ethylene (ET) has a unique effect of inducing N-position epidermal cells to form root hairs. However, the molecular mechanisms underlying ET-induced ectopic root hair development remain enigmatic. Here, we show that ET promotes ectopic root hair formation through down-regulation of GL2 expression. ET-activated transcription factors EIN3 and its homolog EIL1 mediate this regulation. Molecular and biochemical analyses further revealed that EIN3 physically interacts with TTG1 and interferes with the interaction between TTG1 and GL3, resulting in reduced activation of GL2 by the WER-GL3-TTG1 complex. Furthermore, we found through genetic analysis that the master regulator of root hair elongation, RSL4, which is directly activated by EIN3, also participates in ET-induced ectopic root hair development. RSL4 negatively regulates the expression of GL2, likely through a mechanism similar to that of EIN3. Therefore, our work reveals that EIN3 may inhibit gene expression by affecting the formation of transcription-activating protein complexes and suggests an unexpected mutual inhibition between the hair elongation factor, RSL4, and the hair specification factor, GL2. Overall, this study provides a molecular framework for the integration of ET signaling and intrinsic root hair development pathway in modulating root epidermal cell specification.

Salicylic Acid Suppresses Apical Hook Formation via NPR1-Mediated Repression of EIN3 and EIL1 in Arabidopsis.

Salicylic acid (SA) and ethylene (ET) are important phytohormones that regulate numerous plant growth, development, and stress response processes. Previous studies have suggested functional interplay of SA and ET in defense responses, but precisely how these two hormones coregulate plant growth and development processes remains unclear. Our present work reveals antagonism between SA and ET in apical hook formation, which ensures successful soil emergence of etiolated dicotyledonous seedlings. Exogenous SA inhibited ET-induced expression of HOOKLESS1 (HLS1) in Arabidopsis (Arabidopsis thaliana) in a manner dependent on ETHYLENE INSENSITIVE3 (EIN3) and EIN3-LIKE1 (EIL1), the core transcription factors in the ET signaling pathway. SA-activated NONEXPRESSER OF PR GENES1 (NPR1) physically interacted with EIN3 and interfered with the binding of EIN3 to target gene promoters, including the HLS1 promoter. Transcriptomic analysis revealed that NPR1 and EIN3/EIL1 coordinately regulated subsets of genes that mediate plant growth and stress responses, suggesting that the interaction between NPR1 and EIN3/EIL1 is an important mechanism for integrating the SA and ET signaling pathways in multiple physiological processes. Taken together, our findings illuminate the molecular mechanism underlying SA regulation of apical hook formation as well as the antagonism between SA and ET in early seedling establishment and possibly other physiological processes.

Peripheral benzodiazepine receptor TSPO needs to be reconsidered before using as a drug target for a pigmentary disorder.

The peripheral benzodiazepine receptor (TSPO/PBR) is highly conserved among different species but with perplexing biochemical functions. Multiple ligands of TSPO show commendable regulatory activities in lots of biological functions, such as neuro-protection, cholesterol transport, and so on. These researches support that TSPO may be a potential target for disease treatment and drug development. Previous studies have shown that its ligands benzodiazepines show a satisfactory effect on melanogenic promotion. However, the potential application of TSPO in drug development for pigmentary disorder needs further investigation. In this study, we confirmed the melanogenesis induction of TSPO ligand, Ro5-4864 in mouse melanoma cell lines, human skin tissue, and zebrafish embryos by inducing melanin synthesis and melanosome transport. Molecular genetics and pharmacological studies showed that TSPO deficiency did not affect melanin production in B16F10 cells and zebrafish embryos, nor did it affect the melanin promotion effect of Ro5-4864. Whether or not TSPO exists, the expression of lots of melanogenesis-related proteins, such as TYR, TRP-1, DCT, Mlph, and Rab27 was upregulated with the Ro5-4864 administration. These results indicated that Ro5-4864 induces melanogenesis in a TSPO-independent manner, which is inconsistent with previous research. This research is a reminder that we need to be very careful during target validation in drug development.

Research on the Security of IPv6 Communication Based on Petri Net under IoT.

The distribution of wireless network systems challenges the communication security of Internet of Things (IoT), and the IPv6 protocol is gradually becoming the main communication protocol under the IoT. The Neighbor Discovery Protocol (NDP), as the base protocol of IPv6, includes address resolution, DAD, route redirection and other functions. The NDP protocol faces many attacks, such as DDoS attacks, MITM attacks, etc. In this paper, we focus on the communication-addressing problem between nodes in the Internet of Things (IoT). We propose a Petri-Net-based NS flooding attack model for the flooding attack problem of address resolution protocols under the NDP protocol. Through a fine-grained analysis of the Petri Net model and attacking techniques, we propose another Petri-Net-based defense model under the SDN architecture, achieving security for communications. We further simulate the normal communication between nodes in the EVE-NG simulation environment. We implement a DDoS attack on the communication protocol by an attacker who obtains the attack data through the THC-IPv6 tool. In this paper, the SVM algorithm, random forest algorithm (RF) and Bayesian algorithm (NBC) are used to process the attack data. The NBC algorithm is proven to exhibit high accuracy in classifying and identifying data through experiments. Further, the abnormal data are discarded through the abnormal data processing rules issued by the controller in the SDN architecture, to ensure the security of communications between nodes.

To curve for survival: Apical hook development.

Apical hook is a simple curved structure formed at the upper part of hypocotyls when dicot seeds germinate in darkness. The hook structure is transient but essential for seedlings' survival during soil emergence due to its efficient protection of the delicate shoot apex from mechanical injury. As a superb model system for studying plant differential growth, apical hook has fascinated botanists as early as the Darwin age, and significant advances have been achieved at both the morphological and molecular levels to understand how apical hook development is regulated. Here, we will mainly summarize the research progress at these two levels. We will also briefly compare the growth dynamics between apical hook and hypocotyl gravitropic bending at early seed germination phase, with the aim to deduce a certain consensus on their connections. Finally, we will outline the remaining questions and future research perspectives for apical hook development.

Cytokinin regulates apical hook development via the coordinated actions of EIN3/EIL1 and PIF transcription factors in Arabidopsis.

The apical hook is indispensable for protecting the delicate shoot apical meristem while dicot seedlings emerge from soil after germination in darkness. The development of the apical hook is co-ordinately regulated by multiple phytohormones and environmental factors. Yet, a holistic understanding of the spatial-temporal interactions between different phytohormones and environmental factors remains to be achieved. Using a chemical genetic approach, we identified kinetin riboside, as a proxy of kinetin, which promotes apical hook development of Arabidopsis thaliana in a partially ethylene-signaling-independent pathway. Further genetic and biochemical analysis revealed that cytokinin is able to regulate apical hook development via post-transcriptional regulation of the PHYTOCHROME INTERACTING FACTORs (PIFs), together with its canonical roles in inducing ethylene biosynthesis. Dynamic observations of apical hook development processes showed that ETHYLENE INSENSITVE3 (EIN3) and EIN3-LIKE1 (EIL1) are necessary for the exaggeration of hook curvature in response to cytokinin, while PIFs are crucial for the cytokinin-induced maintenance of hook curvature in darkness. Furthermore, these two families of transcription factors display divergent roles in light-triggered hook opening. Our findings reveal that cytokinin integrates ethylene signaling and light signaling via EIN3/EIL1 and PIFs, respectively, to dynamically regulate apical hook development during early seedling development.

Modeling Analysis of SM2 Construction Attacks in the Open Secure Sockets Layer Based on Petri Net.

The detection and defense of malicious attacks are critical to the proper functioning of network security. Due to the diversity and rapid updates of the attack methods used by attackers, traditional defense mechanisms have been challenged. In this context, a more effective method to predict vulnerabilities in network systems is considered an urgent need to protect network security. In this paper, we propose a formal modeling and analysis approach based on Petri net vulnerability exploitation. We used the Common Vulnerabilities and Exposures (CVE)-2021-3711 vulnerability source code to build a model. A patch model was built to address the problems of this model. Finally, the time injected by the actual attacker and the time simulated by the software were calculated separately. The results showed that the simulation time was shorter than the actual attack time, and ultra-real-time simulation could be achieved. By modeling the network system with this method, the model can be found to arrive at an illegitimate state according to the structure of Petri nets themselves and thus discover unknown vulnerabilities. This method provides a reference method for exploring unknown vulnerabilities.

Trusted Data Storage Architecture for National Infrastructure.

National infrastructure is a material engineering facility that provides public services for social production and residents' lives, and a large-scale complex device or system is used to ensure normal social and economic activities. Due to the problems of difficult data collection, long project period, complex data, poor security, difficult traceability and data intercommunication, the archives management of most national infrastructure is still in the pre-information era. To solve these problems, this paper proposes a trusted data storage architecture for national infrastructure based on blockchain. This consists of real-time collection of national infrastructure construction data through sensors and other Internet of Things devices, conversion of heterogeneous data source data into a unified format according to specific business flows, and timely storage of data in the blockchain to ensure data security and persistence. Knowledge extraction of data stored in the chain and the data of multiple regions or fields are jointly modeled through federal learning. The parameters and results are stored in the chain, and the information of each node is shared to solve the problem of data intercommunication.

A Fault-Diagnosis Method for Railway Turnout Systems Based on Improved Autoencoder and Data Augmentation.

In recent years, with the rapid increase in coverage and lines, security maintenance has become one of the top concerns with regard to railway transportation in China. As the key transportation infrastructure, the railway turnout system (RTS) plays a vital role in transportation, which will cause incalculable losses when accidents occur. The traditional fault-diagnosis and maintenance methods of the RTS are no longer applicable to the growing amount of data, so intelligent fault diagnosis has become a research hotspot. However, the key challenge of RTS intelligent fault diagnosis is to effectively extract the deep features in the signal and accurately identify failure modes in the face of unbalanced datasets. To solve the above two problems, this paper focuses on unbalanced data and proposes a fault-diagnosis method based on an improved autoencoder and data augmentation, which realizes deep feature extraction and fault identification of unbalanced data. An improved autoencoder is proposed to smooth the noise and extract the deep features to overcome the noise fluctuation caused by the physical characteristics of the data. Then, synthetic minority oversampling technology (SMOTE) is utilized to effectively expand the fault types and solve the problem of unbalanced datasets. Furthermore, the health state is identified by the Softmax regression model that is trained with the balanced characteristics data, which improves the diagnosis precision and generalization ability. Finally, different experiments are conducted on a real dataset based on a railway station in China, and the average diagnostic accuracy reaches 99.13% superior to other methods, which indicates the effectiveness and feasibility of the proposed method.

Growth asymmetry precedes differential auxin response during apical hook initiation in Arabidopsis.

The development of a hook-like structure at the apical part of the soil-emerging organs has fascinated botanists for centuries, but how it is initiated remains unclear. Here, we demonstrate with high-throughput infrared imaging and 2-D clinostat treatment that, when gravity-induced root bending is absent, apical hook formation still takes place. In such scenarios, hook formation begins with a de novo growth asymmetry at the apical part of a straightly elongating hypocotyl. Remarkably, such de novo asymmetric growth, but not the following hook enlargement, precedes the establishment of a detectable auxin response asymmetry, and is largely independent of auxin biosynthesis, transport and signaling. Moreover, we found that functional cortical microtubule array is essential for the following enlargement of hook curvature. When microtubule array was disrupted by oryzalin, the polar localization of PIN proteins and the formation of an auxin maximum became impaired at the to-be-hook region. Taken together, we propose a more comprehensive model for apical hook initiation, in which the microtubule-dependent polar localization of PINs may mediate the instruction of growth asymmetry that is either stochastically taking place, induced by gravitropic response, or both, to generate a significant auxin gradient that drives the full development of the apical hook.

Eliciting positive emotion through strategic responses to COVID-19 crisis: Evidence from the tourism sector.

While social media are effective means of communicating with adverse customer emotions during a crisis, it remains unclear how tourism organisations can respond to pandemic crisis on social media to prevent negative aftermaths. Using a set-theoretical approach, we investigate how COVID-19 response strategies and linguistic cues of responses are intertwined to evoke positive emotions among consumers. This study entails a qualitative content analysis of tourism organisations' COVID-19 announcements and a social media analytics approach that captures consumers' emotional reactions to these announcements via their Twitter replies. Our results extend some well-established findings in the tourism crisis literature by suggesting that combining innovative response strategy, argument quality, and assertive language can reinforce positive emotions during the COVID-19 crisis. Taking organisational characteristics into consideration, we suggest that young established hotels utilise innovative response strategies, whereas retrenchment response strategies for all types of restaurants should be avoided during the COVID-19 crisis.

Peritumoral Microgel Reservoir for Long-Term Light-Controlled Triple-Synergistic Treatment of Osteosarcoma with Single Ultra-Low Dose.

Local minimally invasive injection of anticancer therapies is a compelling approach to maximize the utilization of drugs and reduce the systemic adverse drug effects. However, the clinical translation is still hampered by many challenges such as short residence time of therapeutic agents and the difficulty in achieving multi-modulation combination therapy. Herein, mesoporous silica-coated gold nanorods (AuNR@SiO2 ) core-shell nanoparticles are fabricated to facilitate drug loading while rendering them photothermally responsive. Subsequently, AuNR@SiO2 is anchored into a monodisperse photocrosslinkable gelatin (GelMA) microgel through one-step microfluidic technology. Chemotherapeutic drug doxorubicin (DOX) is loaded into AuNR@SiO2 and 5,6-dimethylxanthenone-4-acetic acid (DMXAA) is loaded in the microgel layer. The osteosarcoma targeting ligand alendronate is conjugated to AuNR@SiO2 to improve the tumor targeting. The microgel greatly improves the injectability since they can be dispersed in buffer and the injectability and degradability are adjustable by microfluidics during the fabrication. The drug release can, in turn, be modulated by multi-round light-trigger. Importantly, a single super low drug dose (1 mg kg-1 DOX with 5 mg kg-1 DMXAA) with peritumoral injection generates long-term therapeutic effect and significantly inhibited tumor growth in osteosarcoma bearing mice. Therefore, this nanocomposite@microgel system can act as a peritumoral reservoir for long-term effective osteosarcoma treatment.

Interaction between BZR1 and EIN3 mediates signalling crosstalk between brassinosteroids and ethylene.

Plant growth and development are coordinated by multiple environmental and endogenous signals. Brassinosteroid (BR) and ethylene (ET) have overlapping functions in a wide range of developmental processes. However, the relationship between the BR and ET signalling pathways has remained unclear. Here, we show that BR and ET interdependently promote apical hook development and cell elongation through a direct interaction between BR-activated BRASSINOZALE-RESISTANT1 (BZR1) and ET-activated ETHYLENE INSENSITIVE3 (EIN3). Genetic analysis showed that BR signalling is required for ET promotion of apical hook development in the dark and cell elongation under light, and ET quantitatively enhances BR-potentiated growth. BZR1 interacts with EIN3 to co-operatively increase the expression of HOOKLESS1 and PACLOBUTRAZOL RESISTANCE FACTORs (PREs). Furthermore, we found that BR promotion of hook development requires gibberellin (GA), and GA restores the hookless phenotype of BR-deficient materials by activating EIN3/EIL1. Our findings shed light on the molecular mechanism underlying the regulation of plant development by BR, ET and GA signals through the direct interaction of master transcriptional regulators.