Tyrosine phosphorylation of IRF3 by BLK facilitates its sufficient activation and innate antiviral response.

Viral infection triggers the activation of transcription factor IRF3, and its activity is precisely regulated for robust antiviral immune response and effective pathogen clearance. However, how full activation of IRF3 is achieved has not been well defined. Herein, we identified BLK as a key kinase that positively modulates IRF3-dependent signaling cascades and executes a pre-eminent antiviral effect. BLK deficiency attenuates RNA or DNA virus-induced ISRE activation, interferon production and the cellular antiviral response in human and murine cells, whereas overexpression of BLK has the opposite effects. BLK-deficient mice exhibit lower serum cytokine levels and higher lethality after VSV infection. Moreover, BLK deficiency impairs the secretion of downstream antiviral cytokines and promotes Senecavirus A (SVA) proliferation, thereby supporting SVA-induced oncolysis in an in vivo xenograft tumor model. Mechanistically, viral infection triggers BLK autophosphorylation at tyrosine 309. Subsequently, activated BLK directly binds and phosphorylates IRF3 at tyrosine 107, which further promotes TBK1-induced IRF3 S386 and S396 phosphorylation, facilitating sufficient IRF3 activation and downstream antiviral response. Collectively, our findings suggest that targeting BLK enhances viral clearance via specifically regulating IRF3 phosphorylation by a previously undefined mechanism.

Phased Telomere-to-Telomere Reference Genome and Pangenome Reveal an Expansion of Resistance Genes during Apple Domestication.

The cultivated apple (Malus domestica Borkh.) is a cross-pollinated perennial fruit tree of great economic importance. Previous versions of apple reference genomes were unphased, fragmented, and lacked comprehensive insights into the highly heterozygous genome, which impeded genetic studies and breeding programs in apple. In this study, we assembled a haplotype-resolved telomere-to-telomere reference genome for the diploid apple cultivar Golden Delicious. Subsequently, we constructed a pangenome based on twelve assemblies from wild and cultivated apples to investigate different types of resistance gene analogs (RGAs). Our results revealed the dynamics of the gene gain and loss events during apple domestication. Compared with cultivated species, more gene families in wild species were significantly enriched in oxidative phosphorylation, pentose metabolic process, responses to salt, and abscisic acid biosynthesis process. Interestingly, our analyses demonstrated a higher prevalence of RGAs in cultivated apples than their wild relatives, partially attributed to segmental and tandem duplication events in certain RGAs classes. Other types of structural variations, mainly deletions and insertions, have affected the presence and absence of TIR-NB-ARC-LRR (TNL), NB-ARC-LRR (NL), and CC-NB-ARC-LRR (CNL) genes. Additionally, hybridization/introgression from wild species has also contributed to the expansion of resistance genes in domesticated apples. Our haplotype-resolved T2T genome and pangenome provide important resources for genetic studies of apples, emphasizing the need to study the evolutionary mechanisms of resistance genes in apple breeding programs.

The African swine fever virus I10L protein inhibits the NF-κB signaling pathway by targeting IKKß.

Proinflammatory factors play important roles in the pathogenesis of African swine fever virus (ASFV), which is the causative agent of African swine fever (ASF), a highly contagious and severe hemorrhagic disease. Efforts in the prevention and treatment of ASF have been severely hindered by knowledge gaps in viral proteins responsible for modulating host antiviral responses. In this study, we identified the I10L protein (pI10L) of ASFV as a potential inhibitor of the TNF-α- and IL-1ß-triggered NF-κB signaling pathway, the most canonical and important part of host inflammatory responses. The ectopically expressed pI10L remarkably suppressed the activation of NF-κB signaling in HEK293T and PK-15 cells. The ASFV mutant lacking the I10L gene (ASFVΔI10L) induced higher levels of proinflammatory cytokines production in primary porcine alveolar macrophages (PAMs) compared with its parental ASFV HLJ/2018 strain (ASFVWT). Mechanistic studies suggest that pI10L inhibits IKKß phosphorylation by reducing the K63-linked ubiquitination of NEMO, which is necessary for the activation of IKKß. Morever, pI10L interacts with the kinase domain of IKKß through its N-terminus, and consequently blocks the association of IKKß with its substrates IκBα and p65, leading to reduced phosphorylation. In addition, the nuclear translocation efficiency of p65 was also altered by pI10L. Further biochemical evidence supported that the amino acids 1-102 on pI10L were essential for the pI10L-mediated suppression of the NF-κB signaling pathway. The present study clarifies the immunosuppressive activity of pI10L, and provides novel insights into the understanding of ASFV pathobiology and the development of vaccines against ASF. IMPORTANCE African swine fever (ASF), caused by the African swine fever virus (ASFV), is now widespread in many countries and severely affects the commercial rearing of swine. To date, few safe and effective vaccines or antiviral strategies have been marketed due to large gaps in knowledge regarding ASFV pathobiology and immune evasion mechanisms. In this study, we deciphered the important role of the ASFV-encoded I10L protein in the TNF-α-/IL-1ß-triggered NF-κB signaling pathway. This study provides novel insights into the pathogenesis of ASFV and thus contributes to the development of vaccines against ASF.

Evolutionary relationship analysis of STARD gene family and VvSTARD5 improves tolerance of salt stress in transgenic tomatoes.

The steroidogenic acute regulatory protein-related lipid transfer domain (STARD) forms a protein that can bind membrane-derived phospholipid second messengers and plasma membranes. Although it has been reported in many plants, the evolutionary relationship of the STARD gene family has not been systematically analyzed, and functions of the HD-START and HD-START-MEKHLA domain subgroup genes under hormone and abiotic stress are also unclear in grapes. This study identified and analyzed 23 VvSTARD genes, which were distinctly divided into five subgroups according to five conserved domain types. The analyses of codon preference, selective pressure, and synteny relationship revealed that grape had higher homology with Arabidopsis compared with rice. Interestingly, the expression levels of VvSTARD genes in subgroups 1, 2, and 3 exhibited significant upregulation under NaCl treatment at 24 h, but VvSTARD genes in subgroups 4 and 5 were upregulated under methyl jasmonate (MeJA) treatment at 24 h. The subcellular localization showed that VvSTARD5 was localized in the nucleus. Additionally, under NaCl treatment at 24 h, there were an obvious decrease in the relative electrical leakages and the content of malondialdehyde (MDA), while the relative expression level of VvSTARD5 and content of proline were obviously enhanced in three transgenic lines. Therefore, the overexpression of VvSTARD5 greatly increased the salt tolerance of transgenic tomatoes. Collectively, this study preliminarily explores the comprehensive function of the STARD gene family in grapes and verifies the function of VvSTARD5 in response to salt.

New Insights into MdSPS4-Mediated Sucrose Accumulation under Different Nitrogen Levels Revealed by Physiological and Transcriptomic Analysis.

Nitrogen nutrition participates in many physiological processes and understanding the physiological and molecular mechanisms of apple responses to nitrogen is very significant for improving apple quality. This study excavated crucial genes that regulates sugar metabolism in response to nitrogen in apples through physiology and transcriptome analysis, so as to lay a theoretical foundation for improving fruit quality. In this paper, the content of sugar and organic acid in apple fruit at different developmental periods under different nitrogen levels (0, 150, 300, and 600 kg·hm-2) were determined. Then, the transcriptomic analysis was performed in 120 days after bloom (DAB) and 150 DAB. The results showed that the fructose and glucose content were the highest at 120 DAB under 600 kg·hm-2 nitrogen level. Meanwhile, different nitrogen treatments decreased malate content in 30 and 60 DAB. RNA-seq analysis revealed a total of 4537 UniGenes were identified as differentially expressed genes (DEGs) under nitrogen treatments. Among these DEGs, 2362 (52.06%) were up-regulated and 2175 (47.94%) were down-regulated. The gene co-expression clusters revealed that most DEGs were significantly annotated in the photosynthesis, glycolysis/gluconeogenesis, pyruvate metabolism, carbon metabolism, carbon fixation in photosynthetic organisms and plant hormone signal transduction pathways. The key transcription factor genes (ERF, NAC, WRKY, and C2H2 genes) were differentially expressed in apple fruit. Sugar and acid metabolism-related genes (e.g., HXK1, SPS4, SS2, PPC16-2, and MDH2 genes) exhibited significantly up-regulated expression at 120 DAB, whereas they were down-regulated at 150 DAB. Furthermore, the MdSPS4 gene overexpression positively promoted sucrose accumulation in apple callus and fruit. In conclusion, the combinational analysis of transcriptome and the functional validation of the MdSPS4 gene provides new insights into apple responses to different nitrogen levels.

MdbZIP44-MdCPRF2-like-Mdα-GP2 regulate starch and sugar metabolism in apple under nitrogen supply.

Nitrogen (N) is regarded as an essential macronutrient and is tightly associated with carbon (C) metabolism in plants. The transcriptome data obtained from this study showed that the expression level of the apple basic leucine zipper (bZIP) transcription factor (TF) MdbZIP44 was up-regulated in 'Oregon Spur Delicious' (Malus domestica Borkh.) apple fruits under nitrogen supply. MdbZIP44 bound to the promoter of Mdα-GP2 gene and inhibited its expression, thereby promoting starch accumulation and decreasing glucose content in apple and tomato fruits. Besides, overexpression of MdbZIP44 promoted sucrose accumulation by regulating the activities of sucrose metabolism-related enzymes and the expression of sugar metabolism-related genes in apple callus and tomato fruits. Furthermore, biochemical assays indicated that MdbZIP44 directly interacted with MdCPRF2-like, another bZIP gene in apple. Meanwhile, this study found that MdCPRF2-like, along with the MdbZIP44 and MdCPRF2-like complex, could activate the expression of Mdα-GP2, respectively. In conclusion, this study provides a new reference for potential mechanisms underlying that MdbZIP44-MdCPRF2-like-Mdα-GP2 regulates starch and sugar metabolism under nitrogen supply.

Experimental study on the impact of "IDS + JFCS" complex wetting agent on the characteristics of coal bodies.

As China's coal mines have transitioned to deep mining, the ground stress within the coal seams has progressively increased, resulting in reduced permeability and poor wetting ability of conventional wetting agents. Consequently, these agents have become inadequate in fulfilling the requirements for preventing washouts during deep mining operations. In response to the aforementioned challenges, a solution was proposed to address the issues by formulating a composite wetting agent. This composite wetting agent combines a conventional surfactant with a chelating agent called tetrasodium iminodisuccinate (IDS). By conducting a meticulous screening of surfactant monomer solutions, the ideal formulation for the composite wetting agent was determined by combining the monomer surfactant with IDS. Extensive testing, encompassing evaluations of the composite solution's apparent strain, contact angle measurements, and alterations in the oxygenated functional groups on the coal surface, led to the identification of the optimal composition. This composition consisted of IDS serving as the chelating agent and fatty alcohol polyoxyethylene ether (JFCS).Subsequent assessment of the physical and mechanical performance of the coal briquettes treated with the composite wetting agent revealed notable enhancements. These findings signify significant advancements in the field and hold promising implications. Following the application of the composite wetting agent, notable reductions were observed in the dry basis ash and dry basis full sulfur of coal. Additionally, the water content within the coal mass increased significantly, leading to a substantial enhancement in the wetting effect of the coal body. This enhanced wetting effect effectively mitigated the coal body's inclination towards impact, thereby offering technical support for optimizing water injection into coal seams and preventing as well as treating impact ground pressure.

Discovery of electromagnetic polarization in Asian rice wine deterioration process and its applications.

Rice wine, known as yellow wine in China and Japan, possesses considerable nutritional value and holds significant global influence. This study addresses the challenge of preserving rice wine, which is prone to rancidity due to its low alcohol content. Conventional storage techniques employing pottery jars often result in substantial spoilage losses. Through rigorous investigation, this research identifies a polarization phenomenon exhibited by degraded rice wine when subjected to high-frequency microwaves(>60GHz), presenting a pioneering method for detecting spoilage, even within sealed containers. Employing a multi-channel microwave radar apparatus, the study delves into the susceptibility of rice wine to electromagnetic waves across various frequencies, uncovering pronounced polarization traits in deteriorated samples within the E-band microwave spectrum. Furthermore, lab-controlled simulations elucidate a direct correlation between physicochemical alterations and high-frequency Radar Cross Section (RCS) signals during the wine's deterioration process. A novel six-membered Hydrated Cluster hypothesis is proposed, offering insights into the molecular mechanisms underlying this phenomenon. Additionally, dielectric property assessments conducted using vector network analyzers (VNA) reveal noteworthy enhancements in the dielectric constant of deteriorated rice wine, particularly within the high-frequency domain, thereby augmenting detectability. These findings carry implications for refining rice wine preservation techniques and contribute to the advancement of non-destructive testing technologies, enabling the detection of rice wine deterioration or indications thereof, even within sealed vessels.

BLK positively regulates TLR/IL-1R signaling by catalyzing TOLLIP phosphorylation.

TLR/IL-1R signaling plays a critical role in sensing various harmful foreign pathogens and mounting efficient innate and adaptive immune responses, and it is tightly controlled by intracellular regulators at multiple levels. In particular, TOLLIP forms a constitutive complex with IRAK1 and sequesters it in the cytosol to maintain the kinase in an inactive conformation under unstimulated conditions. However, the underlying mechanisms by which IRAK1 dissociates from TOLLIP to activate TLR/IL-1R signaling remain obscure. Herein, we show that BLK positively regulates TLR/IL-1R-mediated inflammatory response. BLK-deficient mice produce less inflammatory cytokines and are more resistant to death upon IL-1ß challenge. Mechanistically, BLK is preassociated with IL1R1 and IL1RAcP in resting cells. IL-1ß stimulation induces heterodimerization of IL1R1 and IL1RAcP, which further triggers BLK autophosphorylation at Y309. Activated BLK directly phosphorylates TOLLIP at Y76/86/152 and further promotes TOLLIP dissociation from IRAK1, thereby facilitating TLR/IL-1R-mediated signal transduction. Overall, these findings highlight the importance of BLK as an active regulatory component in TLR/IL-1R signaling.

Kornblum Oxidation Reaction-Induced Collective Transformation of Lead Polyhalides for Stable Perovskite Photovoltaics.

The iodide vacancy defects generated during the perovskite crystallization process are a common issue that limits the efficiency and stability of perovskite solar cells (PSCs). Although excessive ionic iodides have been used to compensate for these vacancies, they are not effective in reducing defects through modulating the perovskite crystallization. Moreover, these iodide ions present in the perovskite films can act as interstitial defects, which are detrimental to the stability of the perovskite. Here, an effective approach to suppress the formation of vacancy defects by manipulating the coordination chemistry of lead polyhalides during perovskite crystallization is demonstrated. To achieve this suppression, an α-iodo ketone is introduced to undergo a process of Kornblum oxidation reaction that releases halide ions. This process induces a rapid collective transformation of lead polyhalides during the nucleation process and significantly reduces iodide vacancy defects. As a result, the ion mobility is decreased by one order of magnitude in perovskite film and the PSC achieves significantly improved thermal stability, maintaining 82% of its initial power conversion efficiency at 85 °C for 2800 h. These findings highlight the potential of halide ions released by the Kornblum oxidation reaction, which can be widely used for achieving high-performance perovskite optoelectronics.

Comparative population genomics reveals convergent and divergent selection in the apricot-peach-plum-mei complex.

The economically significant genus Prunus includes fruit and nut crops that have been domesticated for shared and specific agronomic traits; however, the genomic signals of convergent and divergent selection have not been elucidated. In this study, we aimed to detect genomic signatures of convergent and divergent selection by conducting comparative population genomic analyses of the apricot-peach-plum-mei (APPM) complex, utilizing a haplotype-resolved telomere-to-telomere (T2T) genome assembly and population resequencing data. The haplotype-resolved T2T reference genome for the plum cultivar was assembled through HiFi and Hi-C reads, resulting in two haplotypes 251.25 and 251.29 Mb in size, respectively. Comparative genomics reveals a chromosomal translocation of ~1.17 Mb in the apricot genomes compared with peach, plum, and mei. Notably, the translocation involves the D locus, significantly impacting titratable acidity (TA), pH, and sugar content. Population genetic analysis detected substantial gene flow between plum and apricot, with introgression regions enriched in post-embryonic development and pollen germination processes. Comparative population genetic analyses revealed convergent selection for stress tolerance, flower development, and fruit ripening, along with divergent selection shaping specific crop, such as somatic embryogenesis in plum, pollen germination in mei, and hormone regulation in peach. Notably, selective sweeps on chromosome 7 coincide with a chromosomal collinearity from the comparative genomics, impacting key fruit-softening genes such as PG, regulated by ERF and RMA1H1. Overall, this study provides insights into the genetic diversity, evolutionary history, and domestication of the APPM complex, offering valuable implications for genetic studies and breeding programs of Prunus crops.

Who is Facing Higher Increased Demand After the Online-Offline Channel Integration? Evidence from Public Hospitals in China.

Background: In recent years, the medical industry has been significantly impacted by internet technology, especially with the growing integration of online and offline channel within public hospitals. Objective: Our study aims to explore the effects of hospitals' online-offline channel integration on doctors' offline visits and investigate how the effects of integration varied across doctors with different professional titles. Methods: Our study employs a panel dataset from a large comprehensive hospital in China and conducts staggered difference-in-differences (DID) approach. Results: We find that online-offline channel integration within public hospitals is associated with about 15.5% increase in offline visits, and the 1% growth of monthly number of online visits is associated with about 10.6% monthly offline visits increase. Furthermore, our results indicate that the effectiveness of online-offline channel integration is more pronounced for doctors with lower professional titles compared to those with higher professional titles. Conclusion: Our study provides evidence for policymakers and hospital managers that integrating online and offline channels can optimize the distribution of medical personnel resources within public hospitals. We recommend that young or less-experienced doctors actively participate in hospital-operated online platforms to enhance their professional skills through practical experience.

Physiological and transcriptome analyses of the effects of excessive water deficit on malic acid accumulation in apple.

Acidity is a determinant of the organoleptic quality of apple, whereas its regulatory mechanism under water stress remains obscure. Fruit from apple 'Yanfu 3' of Fuji trees grown under normal water irrigation (CK), excessive water deficit treatment (DRT) and excessive water irrigation treatment (WAT) were sampled at 85, 100, 115, 130, 145, 160 and 175 days after full bloom designated stages S1, S2, S3, S4, S5, S6 and S7, respectively. DRT treatment reduced the individual fruit weight and fruit moisture content, and increased fruit firmness. The malate content of DRT treatment was higher than that of CK and WAT from stages S1 to S7. RNA sequencing (RNA-seq) analysis of the transcriptome at stages S4, S6 and S7 indicated that malate anabolism was associated with cysteine and methionine, auxin signaling, glyoxylate and dicarboxylate and pyruvate metabolism. Overexpression of MdPEPC4 increased the malate content in apple calli induced by 4% PEG. Our study provides novel insights into the effects of water stress on the molecular mechanism underlying apple fruit acidity.

Grape BES1 transcription factor gene VvBES1-3 confers salt tolerance in transgenic Arabidopsis.

BRI1-EMS-Suppressor 1 (BES1) regulates plant growth, development, and stress resistance, and plays a pivotal role in the brassinosteroid (BR) signal transduction pathway. In this study, a total of 12 BES1 genes were identified in the grape (Vitis vinifera) genome. Phylogenetic, structure, and motif sequence analyses of these genes provided insights into their evolutionary characteristics. Hormone-, stress-, and light-responsive and organ-specific cis-acting elements were identified in VvBES1 gene promoters. Microarray data analysis showed that VvBES1 family members exhibit diverse expression patterns in different organs. Quantitative real-time PCR (qRT-PCR) analysis showed that the expression levels of VvBES1 genes differed in response to BR, methyl jasmonate (MeJA), cold (4 °C), NaCl, and polyethylene glycol (PEG) treatments. The expression of VvBES1-3 was 29-fold higher under salt stress than control at 12 h. Moreover, VvBES1-3-overexpessing Arabidopsis thaliana plants showed lower malondialdehyde content, higher proline content, enhanced antioxidant enzyme (catalase, superoxide dismutase, peroxidase) activities, and higher salt-responsive gene expression levels than wild-type plants under salt stress, indicating that VvBES1-3 overexpression enhances salt stress tolerance in transgenic Arabidopsis. These results will contribute to further understanding the functions of BES1 transcription factors in the abiotic stress response.

Synthesis and evolution of PbS nanocrystals through a surfactant-assisted solvothermal route.

We present a surfactant-assisted solvothermal approach for the controllable synthesis of a PbS nanocrystal at low temperature (85 degrees C). Nanotubes (400 nm in length with an outer diameter of 30 nm), bundle-like long nanorods (about 5-15 mum long and an average diameter of 100 nm), nanowires (5-20 mum in length and with a diameter of 20-50 nm), short nanorods (100-300 nm in length and an axial ratio of 5-10), nanoparticles (25 nm in width with an aspect ratio of 2), and nanocubes (a short axis length of 10 nm and a long axis length of 15 nm) were successfully prepared and characterized by transmission electron microscopy, scanning electron microscopy, and powder X-ray diffraction pattern. A series of experimental results indicated that several experimental factors, such as AOT concentration, ratio of [water]/[surfactant], reaction time, and ratio of the reagents, play key roles in the final morphologies of PbS. Possible formation mechanisms of PbS nanorods and nanotubes were proposed.