WeiTsing, a pericycle-expressed ion channel, safeguards the stele to confer clubroot resistance.

Plant roots encounter numerous pathogenic microbes that often cause devastating diseases. One such pathogen, Plasmodiophora brassicae (Pb), causes clubroot disease and severe yield losses on cruciferous crops worldwide. Here, we report the isolation and characterization of WeiTsing (WTS), a broad-spectrum clubroot resistance gene from Arabidopsis. WTS is transcriptionally activated in the pericycle upon Pb infection to prevent pathogen colonization in the stele. Brassica napus carrying the WTS transgene displayed strong resistance to Pb. WTS encodes a small protein localized in the endoplasmic reticulum (ER), and its expression in plants induces immune responses. The cryoelectron microscopy (cryo-EM) structure of WTS revealed a previously unknown pentameric architecture with a central pore. Electrophysiology analyses demonstrated that WTS is a calcium-permeable cation-selective channel. Structure-guided mutagenesis indicated that channel activity is strictly required for triggering defenses. The findings uncover an ion channel analogous to resistosomes that triggers immune signaling in the pericycle.

Mechanistic insights into phosphoactivation of SLAC1 in guard cell signaling.

Stomata in leaves regulate gas (carbon dioxide and water vapor) exchange and water transpiration between plants and the atmosphere. SLow Anion Channel 1 (SLAC1) mediates anion efflux from guard cells and plays a crucial role in controlling stomatal aperture. It serves as a central hub for multiple signaling pathways in response to environmental stimuli, with its activity regulated through phosphorylation via various plant protein kinases. However, the molecular mechanism underlying SLAC1 phosphoactivation has remained elusive. Through a combination of protein sequence analyses, AlphaFold-based modeling and electrophysiological studies, we unveiled that the highly conserved motifs on the N- and C-terminal segments of SLAC1 form a cytosolic regulatory domain (CRD) that interacts with the transmembrane domain(TMD), thereby maintaining the channel in an autoinhibited state. Mutations in these conserved motifs destabilize the CRD, releasing autoinhibition in SLAC1 and enabling its transition into an activated state. Our further studies demonstrated that SLAC1 activation undergoes an autoinhibition-release process and subsequent structural changes in the pore helices. These findings provide mechanistic insights into the activation mechanism of SLAC1 and shed light on understanding how SLAC1 controls stomatal closure in response to environmental stimuli.

Structure and activity of SLAC1 channels for stomatal signaling in leaves.

Stomata in leaves regulate gas exchange between the plant and its atmosphere. Various environmental stimuli elicit abscisic acid (ABA); ABA leads to phosphoactivation of slow anion channel 1 (SLAC1); SLAC1 activity reduces turgor pressure in aperture-defining guard cells; and stomatal closure ensues. We used electrophysiology for functional characterizations of Arabidopsis thaliana SLAC1 (AtSLAC1) and cryoelectron microscopy (cryo-EM) for structural analysis of Brachypodium distachyon SLAC1 (BdSLAC1), at 2.97-Å resolution. We identified 14 phosphorylation sites in AtSLAC1 and showed nearly 330-fold channel-activity enhancement with 4 to 6 of these phosphorylated. Seven SLAC1-conserved arginines are poised in BdSLAC1 for regulatory interaction with the N-terminal extension. This BdSLAC1 structure has its pores closed, in a basal state, spring loaded by phenylalanyl residues in high-energy conformations. SLAC1 phosphorylation fine-tunes an equilibrium between basal and activated SLAC1 trimers, thereby controlling the degree of stomatal opening.

PKCδ serves as a potential biomarker and therapeutic target for microglia-mediated neuroinflammation in Alzheimer's disease.

INTRODUCTION: To investigate the role of a novel type of protein kinase C delta (PKCδ) in the neuroinflammation of Alzheimer's disease (AD). METHODS: We analyzed PKCδ and inflammatory cytokines levels in cerebrospinal fluid (CSF) of AD and normal controls, as well as their correlations. The cellular expression pattern of PKCδ and the effects of PKCδ modulation on microglia-mediated neuroinflammation were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR), western blot, RNA sequencing (RNA-seq), and immunofluorescence staining. RESULTS: PKCδ levels were increased dramatically in the CSF of AD patients and positively correlated with cytokines. PKCδ is expressed mainly in microglia in the brain. Amyloid beta (Aß) stimulation increased PKCδ expression and secretion, which led to upregulation of the nuclear factor kappa B (NF-κB) pathway and overproduction of proinflammatory cytokines. Downregulation or inhibition of PKCδ attenuated Aß-induced microglial responses and improved cognitive function in an AD mouse model. DISCUSSION: Our study identifies PKCδ as a potential biomarker and therapeutic target for microglia-mediated neuroinflammation in AD. HIGHLIGHTS: Protein kinase C delta (PKCδ) levels increase in cerebrospinal fluid (CSF) of patients with Alzheimer's disease (AD), and positively correlate with elevated inflammatory cytokines in human subjects. PKCδ is expressed mainly in microglia in vivo, whereas amyloid beta (Aß) stimulation increases PKCδ expression and secretion, causing upregulation of the nuclear factor kappa B (NF-κB) pathway and production of inflammatory cytokines. Downregulation or inhibition of PKCδ attenuates Aß-enhanced NF-κB signaling and cytokine production in microglia and improves cognitive function in AD mice. PKCδ serves as a potential biomarker and therapeutic target for microglia-mediated neuroinflammation in AD.

If They Won't Know, I Won't Wait: Anticipated Social Consequences Drive Children's Performance on Self-Control Tasks.

This research evaluated the hypothesis that the act of offering an incentive produces anticipated social benefits that are distinct from the benefits associated with the incentive itself. Across three preregistered studies, 3- to 5-year-old children in China (total N = 210) were given an opportunity to wait for an additional sticker (Studies 1 and 3) or an edible treat (Study 2). Rewards were dispensed via a timer-controlled box that allowed the experimenter's apparent ability to learn how long children waited to be manipulated experimentally. Children waited only about half as long when they believed the experimenter would not find out how long they waited. When children were offered three prizes for waiting, anticipated social benefits still drove behavior at least as much as the reward. The findings demonstrate that children as young as 3 years are sensitive to anticipated social rewards when responding to offers of incentives.

Serum total bilirubin concentration is associated with carotid atherosclerosis in patients with prehypertension.

Objectives: Bilirubin has been demonstrated to be linked with anti-inflammatory and antioxidant progress. We aimed to evaluate the association between serum total bilirubin level and carotid intima-media thick-ness (cIMT) in patients with prehypertension. METHODS: We consecutively enrolled pre-hypertensive patients from a community in Guangzhou between January 2017 and January 2018. All patients underwent carotid artery ultrasonography measurement. The correlation between serum total bilirubin and cIMT was assessed by using the Pearson's correlation coefficient. Multiple logistic regression analysis was performed to assess the independent association between clinical parameters and carotid atherosclerosis. RESULTS: A total of 691 subjects with prehypertension were included in this study. There were 101 patients with increased cIMT (mean age 52.69 ± 11.58 years; 50 male) and 590 subjects with normal cIMT (mean age50.28 ± 10.33 years; 332 male). We found that cIMT was significantly related with systolic blood pressure(r = 0.257, P < 0.001), C-reactive protein (r = 0.327, P < 0.001), total cholesterol (r = 0.218, P = 0.002) and total bilirubin (r =-0.489, P < 0.001). A multiple logistic regression analysis revealed that total bilirubin was an independent factor for atherosclerosis (OR = 0.476; 95%CI: 0.253, 0.764; P < 0.001). CONCLUSION: Our results suggested that serum total bilirubin was inversely related with cIMT, and might be an early clinical marker for predicting the occurrence of subclinical carotid atherosclerosis in patients with prehypertension.

Effects of intensive blood-pressure treatment on myocardial work in elderly hypertensive patients: A subcenter study of the STEP randomized controlled trial.

BACKGROUND: The benefits and safety of intensive blood pressure treatment in elderly hypertensive patients have been proved in the STEP trial. However, relevant mechanisms for intensive treatment are lacking. HYPOTHESIS: We aimed to explore whether intensive blood pressure treatment is associated with left ventricular systolic function changes as evaluated by myocardial work (MW) parameters in elderly hypertensive patients compared to the standard. METHODS: Patients were randomized to the intensive group (n = 66, median age 66 years, 42.4% male) with a systolic blood pressure (SBP) goal of 110 to <130 mmHg or the standard treatment group (n = 50, median age 63.5 years, 30% male) with an SBP goal of 130-<150 mmHg in this subcenter study of the STEP trial. There was no pre-randomization echocardiographic collected. Echocardiographic exam was produced at 1-year (phase 1) and 3-year (phase 2) post-randomization. RESULTS: In phase 1, SBP was already significantly lower in the intensive treatment group than in the standard treatment group (126.5 vs. 132.1 mmHg, p < .05). During a median follow-up of 40 months, in phase 2, the intensive group still had a lower SBP than the standard treatment group (125.0 vs. 135.3 mmHg, p < .05). Both global work index (GWI) and global constructive work (GCW) decreased significantly in phase in the intensive treatment group but not in the standard group (p < .05). Global wasted work (GWW) increased and global work efficiency (GWE) declined in both groups from phase 1 to phase 2 while no significant difference between the treatment effects. Similarly, left ventricular ejection function (LVEF) and global longitudinal strain (GLS) decreased in the two groups. The multivariate linear regression analysis showed the intensive treatment appeared to be an independent predictor of the ΔGWI (ß = -110.92; 95% CI, -197.78 to -30.07, p = .008) and ΔGCW (ß = -135.11; 95% CI, -220.33 to -49.88, p = .002). CONCLUSIONS: In elderly hypertensive patients, lower SBP was associated with decreased GWI and GCW and intensive BP treatment did not improve global MW efficiency.

Structure and activation mechanism of the rice Salt Overly Sensitive 1 (SOS1) Na+/H+ antiporter.

Salinity is one of the most severe abiotic stresses that adversely affect plant growth and agricultural productivity. The plant Na+/H+ antiporter Salt Overly Sensitive 1 (SOS1) located in the plasma membrane extrudes excess Na+ out of cells in response to salt stress and confers salt tolerance. However, the molecular mechanism underlying SOS1 activation remains largely elusive. Here we elucidate two cryo-electron microscopy structures of rice (Oryza sativa) SOS1, a full-length protein in an auto-inhibited state and a truncated version in an active state. The SOS1 forms a dimeric architecture, with an NhaA-folded transmembrane domain portion in the membrane and an elongated cytosolic portion of multiple regulatory domains in the cytoplasm. The structural comparison shows that SOS1 adopts an elevator transport mechanism accompanied by a conformational transition of the highly conserved Pro148 in the unwound transmembrane helix 5 (TM5), switching from an occluded conformation in the auto-inhibited state to a conducting conformation in the active state. These findings allow us to propose an inhibition-release mechanism for SOS1 activation and elucidate how SOS1 controls Na+ homeostasis in response to salt stress.

Cryo-EM structure and electrophysiological characterization of ALMT from Glycine max reveal a previously uncharacterized class of anion channels.

Aluminum-activated malate transporters (ALMTs) form an anion channel family that plays essential roles in diverse functions in plants. Arabidopsis ALMT12, also named QUAC1 (quick anion channel 1), regulates stomatal closure in response to environmental stimuli. However, the molecular basis of ALMT12/QUAC1 activity remains elusive. Here, we describe the cryo-EM structure of ALMT12/QUAC1 from Glycine max at 3.5-Å resolution. GmALMT12/QUAC1 is a symmetrical dimer, forming a single electropositive T-shaped pore across the membrane. The transmembrane and cytoplasmic domains are assembled into a twisted two-layer architecture, with their associated dimeric interfaces nearly perpendicular. GmALMT12/QUAC1-mediated currents display rapid kinetics of activation/deactivation and a bell-shaped voltage dependency, reminiscent of the rapid (R)-type anion currents. Our structural and functional analyses reveal a domain-twisting mechanism for malate-mediated activation. Together, our study uncovers the molecular basis for a previously uncharacterized class of anion channels and provides insights into the gating and modulation of the ALMT12/QUAC1 anion channel.