Reverse Reaction Research Articles

PP2C phosphatase Pic6 suppresses MAPK activation and disease resistance in tomato.

Type 2C protein phosphatases (PP2Cs) are essential for regulating plant immune responses to pathogens. Our study focuses on the tomato PP2C-immunity associated candidate 6 (Pic6), elucidating its role in negatively regulating pattern-triggered immunity (PTI) signaling pathways in tomato. Using reverse transcription quantitative polymerase chain reaction (RT-qPCR), we observed that treatment with microbe-associated molecular patterns (MAMPs)- flg22 and flgII-28-significantly increased Pic6 mRNA levels in wild-type (RG-PtoR) tomato plants. Pic6 features a conserved N-terminal kinase-interacting motif (KIM) and a C-terminal PP2C domain. We produced variants of Pic6 with mutations in these regions, demonstrating their involvements in negatively regulating tomato immunity. Agrobacterium-mediated transient overexpression of Pic6 resulted in enhanced growth of the bacterial pathogen Pseudomonas syringae pv. tomato (Pst) strain DC3000ΔhopQ1-1 compared to a YFP control. Additionally, Pic6 overexpression inhibited mitogen-activated protein kinase (MAPK) activation in response to flg22 and flgII-28 treatments. Importantly, Pic6 exhibited phosphatase activity and interacted with tomato Mkk1/Mkk2 proteins and dephosphorylated them in a KIM-dependent manner. Furthermore, we generated RG-pic6 loss-of-function mutants by CRISPR/Cas9, revealing that the absence of Pic6 heightened MAPK activity and increased resistance to Xanthomonas euvesicatoria strain 85-10 (Xe 85-10) when compared with the wild-type (RG-PtoR) plants. Transcript analyses showed that after flg22/flgII-28 treatment, PTI-reporter genes NAC and osmotin were significantly upregulated in RG-pic6 mutants in comparison to the wild-type (RG-PtoR) plants. Overall, our findings indicate that Pic6 acts as a negative regulator of MAPK signaling and playing a pivotal role in modulating tomato immunity against bacterial pathogens.

Increased caveolin 1 by human antigen R exacerbates Porphyromonas gingivali-induced atherosclerosis by modulating oxidative stress and inflammatory responses

Objective: Many different types of infectious oral diseases have been identified clinically, including chronic periodontitis. Porphyromonas gingivalis is the main pathogen causing chronic periodontitis, which is closely related to atherosclerosis (AS) and can promote the expression levels of caveolin 1 (Cav-1) and induced ribonucleic acid (RNA)-binding protein human antigen R (HuR). However, the roles of Cav-1 and its relationship with HuR in P. gingivalis-mediated AS progression remain largely unknown. Here, we aimed to detect the role and molecular mechanisms of Cav-1 in P. gingivalis-mediated AS. Material and Methods: To investigate the role of Cav-1 in P. gingivalis-mediated AS, we infected human umbilical vein endothelial cells (HUVECs) with P. gingivalis at a multiplicity of infection of 100:1 for 6, 12, and 24 h to simulate P. gingivalis-induced AS models in vitro and then transfected them with Cav-1 small interfering RNA to silence Cav-1. Combining molecular biology experimental techniques such as cell counting kit-8 assay, enzyme-linked immunosorbent assay, immunofluorescence staining, flow cytometry, Western blotting, and Oil Red O staining, and apolipoprotein E-deficient AS model mice, the impacts of Cav-1 on cell viability, inflammation, oxidative stress, apoptosis, Cav-1 and intercellular cell adhesion molecule-1 (ICAM-1) levels, and atherosclerotic plaque formation were investigated. Then, the relationship between Cav-1 and HuR was investigated through biotin pull-down and RNA immunoprecipitation assays, reverse transcription quantitative polymerase chain reaction, and Western blot. Results: P. gingivalis can induce Cav-1 expression in a time- and dose-dependent manner (P < 0.05). This effect can inhibit the proliferation of HUVECs (P < 0.05). Cav-1 interference repressed inflammatory response, reactive oxygen species (ROS) and ICAM-1 levels, and apoptosis in the HUVECs (P < 0.05). Cav-1 messenger RNA was stabilized by HuR, which can bind to the 3’ untranslated region of Cav-1. Increase in HuR level reversed the effects of Cav-1 silencing on ROS and ICAM-1 levels and apoptosis in the HUVECs (P < 0.05). In addition, the levels of inflammatory response, oxidative stress, and atherosclerotic plaque formation induced by P. gingivalis in the mouse model were significantly reduced after Cav-1 expression was inhibited (P < 0.05). Conclusion: HuR-activated Cav-1 may promote atherosclerotic plaque formation by modulating inflammatory response and oxidative stress, leading to AS.

Temperature-Sensitive Magnetic Resonance Probes: Leveraging Hyperpolarized Pyridine-2-Carbaldehyde and SABRE for Real-Time Temperature Sensing.

In this study, we developed a novel approach for real-time, temperature-sensitive nuclear magnetic resonance (NMR) measurements using signal amplification by reversible exchange (SABRE). We discovered that pyridine-2-carbaldehyde (A) exhibits different behavior depending on temperature, showing high hyperpolarization efficiency. In contrast, its reversible reaction product, the hemiacetal form (A'), is not affected by temperature. Exploiting this difference, we achieved a reliable polarization enhancement ratio without internal standard materials, even at low concentrations. Our method overcomes challenges associated with SABRE for 2-functionalized pyridines, enabling direct temperature monitoring in real-time NMR studies. We successfully applied it to monitor temperatures in solutions containing SABRE-inactive compounds like caffeine. Furthermore, we demonstrated its efficacy using a 60 MHz benchtop NMR spectrometer, validating its potential in challenging environments where conventional techniques may be limited. This technique shows promise for influencing the magnetic resonance field, potentially facilitating more accurate real-time analyses of molecular reactions and structures. Future research will focus on adapting this method for biological settings, aiming to stimulate advancements in NMR and magnetic resonance imaging (MRI) applications.

Anti-inflammatory role of low-intensity pulsed ultrasound in inhibiting lipopolysaccharide-induced M1 polarization of RAW264.7 cells via Wnt2b/AXIN/β-catenin

Background Low-intensity pulsed ultrasound (LIPUS) is a special type of low-intensity ultrasound. In periodontal disease, LIPUS is applied as an adjuvant and non-invasive treatment. It has been reported that LIPUS significantly shifts the macrophage phenotype from M1 to M2, but the specific mechanism behind this shift is still unknown. Methods RAW264.7 cells were induced to M1/M2 polarization with lipopolysaccharide (LPS)/interleukin-4 (IL4). LIPUS was performed for 25 min two times, 24 h apart, at an intensity of 45 mW/cm2 to stimulate RAW264.7 cells. PolyA mRNA sequencing was conducted of both the LPS-induced RAW264.7 cells and the LPS-induced RAW264.7 cells with LIPUS treatment. The expression of Wnt2b in RAW264.7 cells was downregulated by siRNA. The macrophage surface markers and downstream inflammatory cytokines were detected using flow cytometry. The relative expression of proteins in the Wnt2b/AXIN/β-catenin pathway was assessed using reverse transcription real-time polymerase chain reaction (RT-qPCR) and Western blot. Results LIPUS reversed the M1 polarization of RAW264.7 cells, with decreased expression of CD80 and CD86. In addition, LIPUS enhanced the M2 polarization of RAW264.7 cells, with upregulated expression of CD163 and CD206. The polyA mRNA sequencing results indicated that the Wnt signaling pathway participated in the M1 polarization of LIPUS-treated RAW264.7. The results of the RT-qPCR showed a higher expression of Wnt2b in LIPUS-treated and M1- or M2-polarized RAW264.7 cells. Knocking down Wnt2b was shown to reverse the inhibitory effect of LIPUS on M1 polarization and increase the expression of CD80 and CD86. Wnt2b knockdown also regulated downstream AXIN, β-catenin, and inflammatory factors such as tumor necrosis factor alpha (TNFα) and interleukin-6 (IL6). Conclusions LIPUS plays an anti-inflammatory role by inhibiting LPS-induced M1 polarization of RAW264.7 cells in a Wnt2b/AXIN/β-catenin-dependent way. LIPUS may play a therapeutic role in periodontal diseases by inhibiting inflammation through the regulation of macrophage differentiation.

Advances in Self-Healing Perovskite Solar Cells Enabled by Dynamic Polymer Bonds.

This comprehensive review addresses the self-healing phenomenon in perovskite solar cells (PSCs), emphasizing the reversible reactions of dynamic bonds as the pivotal mechanism. The crucial role of polymers in both enhancing the inherent properties of perovskite and inducing self-healing phenomena in grain boundaries of perovskite films are exhibited. The review initiates with an exploration of the various stability problems that PSCs encounter, underscoring the imperative to develop PSCs with extended lifespans capable of self-heal following damage from moisture and mechanical stress. Owing to the strong compatibility brought by polymer characteristics, many additive strategies can be employed in self-healing PSCs through artful molecular design. These strategies aim to limit ion migration, prevent moisture ingress, alleviate mechanical stress, and enhance charge carrier transport. By scrutinizing the conditions, efficiency, and types of self-healing behavior, the review encapsulates the principles of dynamic bonds in the polymers of self-healing PSCs. The meticulously designed polymers not only improve the lifespan of PSCs through the action of dynamic bonds but also enhance their environmental stability through functional groups. In addition, an outlook on self-healing PSCs is provided, offering strategic guidance for future research directions in this specialized area.

Melatonin attenuates cardiac dysfunction and inflammation in dilated cardiomyopathy via M2 macrophage polarization.

Melatonin is a neuroendocrine hormone that exerts protective effects on the heart. Increasing evidence suggests that macrophage M2-type polarization improves myocardial regeneration and repair. Therefore, this study investigated whether melatonin ameliorates dilated cardiomyopathy (DCM) by modulating M2-type polarization. DCM mice were established by induction with doxorubicin and then treated with melatonin. Cardiac dysfunction was determined by measuring left ventricular ejection fraction (LVEF) and left ventricular internal dimensions at end-diastole and end-systole. Heart injury and fibrosis were determined by hematoxylin and eosin staining and Sirius Red staining, respectively. Serum concentrations of melatonin, tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6) were measured through enzyme-linked immunosorbent assays. M2-type macrophages were analyzed by flow cytometry. Relative mRNA and protein levels were determined by reverse transcription quantitative polymerase chain reaction and western blotting, respectively. Circulating melatonin levels were significantly decreased in DCM mice and were associated with LVEF. Treatment with melatonin markedly ameliorated cardiac dysfunction, improved survival, and alleviated pathological changes and collagen deposition in DCM mice. Furthermore, melatonin-treated DCM mice displayed lower serum and cardiac levels of IL-1β, IL-6, and TNF-α, as well as higher numbers of M2-type macrophages in cardiac tissue, indicating that melatonin treatment could decrease inflammatory responses and facilitate M2 macrophage polarization in DCM mice. Thus, melatonin treatment alleviated cardiac dysfunction and inflammatory responses by promoting M2 macrophage polarization in the DCM mouse model.

An epidemiologic surveillance study based on wastewater and respiratory specimens reveals influenza a virus prevalence and mutations in Taiyuan, China during 2023-2024.

Influenza is a major cause of morbidity and mortality. Influenza A virus (IAV) is one of the most important pathogens causing influenza and often causes global pandemics due to its tendency to mutate. We aim to use epidemiology based on wastewater and respiratory specimens to understand the occurrence of influenza A virus infections in Taiyuan City. A retrospective epidemiology surveillance was carried out at the First Hospital of Shanxi Medical University (FHSMU) and five wastewater treatment plants (WTPs) in Taiyuan city from 2023 to 2024. Reverse transcription real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) was used to detect influenza A viruses in wastewater and respiratory specimens. High-throughput whole genome sequencing was performed on 17 strains obtained in this study, and subsequent analyses included characterization, phylogenetic construction, amino acid mutation analysis, and antigenic structural variability assessment. 520 wastewater samples and 1,203 throat swab samples were collected. We detected RNA concentration from pH1N1 and H3N2 viruses in wastewater and got 17 genome sequences (5 of pH1N1 and 12 of H3N2) in respiratory specimens. Whole-genome sequencing showed co-prevalence of pH1N1 viruses in the branches of 6B.1A.5a.2a.1 and H3N2 viruses in the branches of 3C.2a1b.2a.2a.3a.a in Taiyuan from 2023 to 2024. Moreover, a HA mutation (N138D), predicted to be of high phenotypic consequence, was found in 8 Taiyuan H3N2 sequences. This study highlights the predominant presence of pH1N1 and H3N2 strains in Taiyuan. The analysis also identified amino acid site variations in the HA antigenic epitopes in H3N2 strains, which may contribute to immune escape.

Recruitment and polarization typing of tumor-associated macrophages is associated with tumor progression and poor prognosis in Wilms tumor patients.

Tumor-associated macrophages (TAMs) play a crucial role in shaping various tumor microenvironments. However, their recruitment in Wilms tumor (WT), the predominant malignant renal tumor in children, has been inadequately explored. This retrospective cohort study involved the analysis of 148 WT samples to investigate the recruitment and polarization typing of TAMs in WT tissues. WT tissues underwent Western blotting (WB), reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and immunofluorescence (IF) to measure the expression of TAM markers CD68, CD86, and CD163. Statistically analyze the relationship between TAM recruitment levels and patient clinical characteristics, and use Kaplan-Meier curves and the log-rank test to evaluate the association between TAM levels and survival outcomes. The findings indicated a positive correlation between the recruitment levels of total macrophages (Mtotal) and M2 tumor-associated macrophages (M2 TAM) in both chemotherapy and non-chemotherapy groups with the clinical stage. Elevated recruitment of Mtotal and M2 TAM in tumor tissues was linked to a poorer prognosis. Notably, patients with persistently higher recruitment of M2 TAM following preoperative chemotherapy exhibited the worst prognosis. The recruitment and polarization typing of TAM exhibit significant differences in WT patients with various stages and prognosis outcomes, suggesting a potential avenue for future diagnosis and treatment of WT.

A pyroptosis-related lncRNA risk model for the prediction of prognosis and immunotherapy response in head and neck squamous cell carcinoma

BackgroundRecent research has highlighted pyroptosis as a key factor in cancer progression. This study aims to explore the association between pyroptosis-related signatures and overall survival (OS) in head and neck squamous cell carcinoma (HNSC) and develop a pyroptosis-related long non-coding RNA (lncRNA) risk model to predict prognosis and response to immunotherapy in HNSC.MethodsWe extracted expression data for 18 pyroptosis-related genes and identified lncRNA probes specific to HNSC by using datasets from the Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA). Consensus clustering was performed to categorize HNSC patients into distinct subtypes. A six-lncRNA risk score model was constructed through univariate and least absolute shrinkage and selection operator (LASSO) Cox regression analyses. We evaluated the predictive ability of the lncRNA model for patients’ survival and immunotherapy response. Gene expression was evaluated using immunohistochemistry (IHC) and Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR).ResultsOur analysis revealed two distinct pyroptosis-related subtypes in HNSC patients, Cluster A and Cluster B. Notably, patients in Cluster B exhibited significantly poorer overall survival compared to those in Cluster A. Through differential expression analysis, we identified six lncRNAs (AC002331.1, CTA-384D8.35, RP11-291B21.2, AC006262.5, RP1-27K12.2, and RP11-54H7.4) that were differentially expressed between these clusters. A 6-lncRNA risk score model was developed, which successfully stratified patients into high- and low-risk groups with distinct overall survival outcomes. Validation using RT-qPCR confirmed the differential expression of these six lncRNAs in HNSC tumor tissues compared to adjacent normal tissues, we found that the expression of CTA-384D8.35 was significantly increased in the tumor group (t=-6.203, P<0.001). Furthermore, the 6-lncRNA risk score model demonstrated a significant association with patient response to immunotherapy, with the low-risk group exhibiting a higher objective response rate to immune checkpoint blockade (ICB) therapy and longer survival compared to the high-risk group.ConclusionOur study underscores the role of pyroptosis signatures in HNSC prognosis and identifies two distinct pyroptosis subtypes with differing survival outcomes. The six-lncRNA risk score model offers a valuable tool for predicting patient prognosis and potential benefits from ICB therapy. These findings highlight the importance of pyroptosis and associated lncRNAs in the tumor microenvironment, paving the way for novel targeted therapies in HNSC.

Exploring lncRNA expression in follicular fluid exosomes of patients with obesity and polycystic ovary syndrome based on high-throughput sequencing technology.

Infertility is a reproductive health problem that attracts worldwide attention. Polycystic ovary syndrome (PCOS) is a major cause of female infertility and patients with obesity and PCOS are particularly common in clinical practice. Long non-coding RNA (lncRNAs) are a functional core in cells that regulate gene expression, transcription, and chromatin modification processes, and participate in epigenetics, cell cycle, and cell differentiation. LncRNAs are assumed to play a role in the occurrence and development of PCOS; however, their specific mechanism of action remains to be elucidated. High-throughput sequencing technology has been used to sequence and analyze lncRNAs in exosomes from the follicular fluid of patients with obesity and PCOS and those who underwent assisted reproductive therapy owing to male factors. Specific expression profiles of patients with obesity and PCOS were obtained and functional information analysis combined with a literature review were performed to screen for differentially expressed lncRNAs, which were validated using real-time reverse transcription quantitative polymerase chain reaction (RT-qPCR). High-throughput sequencing analysis revealed that compared to normal patients with male infertility, patients with obesity and PCOS had a total of 20 lncRNAs with significant expression differences in follicular fluid exosomes. Among them, 17 lncRNAs were upregulated and three were downregulated. Functional analysis showed that differentially expressed genes were mainly enriched in "cell metabolism," "cell adhesion," and other aspects: related gene pathways mainly involved Huntington's disease, Parkinson's disease, spliceosomes, non-alcoholic fatty liver disease, and ribosomes. Verification of differentially expressed lncRNAs revealed that the expression of lncRNAs TPT1-AS1, PTOV1-AS1, PTPRG-AS1, and SNHG14 in follicular fluid exosomes was consistent with the sequencing results. A preliminary differential expression profile of lncRNAs in exosomes of patients with obesity and PCOS was established by transcriptomic analysis of these individuals. Our bioinformatics analysis results may be applicable to further study of the impact mechanism involving obesity and PCOS. These differentially expressed lncRNAs maybe served as potential biomarkers for in-depth studies of the occurrence, development on Follicle quality and function for patients with PCOS in the future.

Crystallographic Dimensionality Determines the Electrochemical Reaction Mechanism in Alkali Transition-Metal Chlorides.

Intense research efforts on transition metal chalcogenides (oxides and sulfides), pnictides (nitrides and phosphides), and fluorides have demonstrated the complex, intertwined effects of structural and chemical changes on their electrochemical response leading to intercalation, conversion, or displacement reactions when reacting with lithium. Prior efforts largely left halides unexplored due to their heightened solubility in classical liquid electrolytes. In this work, we employ superconcentrated electrolytes to demonstrate the composition- and structure-dependent electrochemical reactivity of A2MCl4 compounds (A = Li or Na and M = Cr, Mn, Fe, and Co). Comparing four lithiated compounds, we demonstrate that they all undergo conversion reactions when reacting with 2 Li+ per formula unit, associated with large polarization and limited cycling ability. Nevertheless, combining in situ XRD with post-mortem XPS and STEM/EDS analysis, we demonstrate that Li2CoCl4 first reacts with one Li+ following a displacement reaction providing a reversible capacity of 125 mAh g-1. This reaction is enabled by the formation of a Li6CoCl8 intermediate, which shares a similar anionic framework with pristine Li2CoCl4, ensuring the topotactic insertion of Li+ balanced by the Co2+/Co0 redox couple and the formation of metallic Co nanoparticles. Comparing these compounds, we propose that two criteria are necessary to trigger the displacement reaction in A2MCl4 compounds: the presence of 1D chains of edge-sharing octahedra to favor electronic delocalization and the availability of a metal-deficient intermediate. Screening numerous A2MCl4 compounds, we demonstrate the universality of these design principles, which extend to Na-ion materials by demonstrating a low-polarization, reversible displacement reaction for Na2MnCl4.

Interfacial Proton Precompensation: Suppressing Deprotonation-Driven Lattice Collapse for Enhanced Efficiency and Stability in Perovskite Solar Cells.

The chemical property of the buried interface plays a crucial role in improving the performance and stability of perovskite solar cells (PSCs). The SnO2/perovskite interface prepared from SnO2 alkaline hydrogel with high proton affinity triggers directional migration and irreversible reactions of protons, exacerbating the disintegration of perovskite crystal. In this study, we proposed proton precompensation strategy to suppress the deprotonation effect of the buried interface and improve the durability of the devices. By modulating the chemical environment and surface energy state of the buried interface, the domain-limiting and spontaneous compensation of protons in formamidinium (FA+) under Coulomb force were achieved, thereby stabilizing the perovskite crystal structure. The stability of target perovskite films under UV illumination and heating at 85 °C was significantly enhanced. As a result, the devices can retain around 90% of their initial power conversion efficiency (PCE) after 1000 h of continuous irradiation at the maximum power point (MPP). Moreover, due to the reduction of defect content at the buried interface and the improvement of conductivity and carrier mobility by the precompensation treatment, the interfacial energy loss and non-radiative recombination were substantially diminished. The target PSC devices exhibited much higher PCE of 25.55% than the control devices (23.03%).

Interfacial Proton Precompensation: Suppressing Deprotonation‐Driven Lattice Collapse for Enhanced Efficiency and Stability in Perovskite Solar Cells

The chemical property of the buried interface plays a crucial role in improving the performance and stability of perovskite solar cells (PSCs). The SnO2/perovskite interface prepared from SnO2 alkaline hydrogel with high proton affinity triggers directional migration and irreversible reactions of protons, exacerbating the disintegration of perovskite crystal. In this study, we proposed proton precompensation strategy to suppress the deprotonation effect of the buried interface and improve the durability of the devices. By modulating the chemical environment and surface energy state of the buried interface, the domain‐limiting and spontaneous compensation of protons in formamidinium (FA+) under Coulomb force were achieved, thereby stabilizing the perovskite crystal structure. The stability of target perovskite films under UV illumination and heating at 85 °C was significantly enhanced. As a result, the devices can retain around 90% of their initial power conversion efficiency (PCE) after 1000 h of continuous irradiation at the maximum power point (MPP). Moreover, due to the reduction of defect content at the buried interface and the improvement of conductivity and carrier mobility by the precompensation treatment, the interfacial energy loss and non‐radiative recombination were substantially diminished. The target PSC devices exhibited much higher PCE of 25.55% than the control devices (23.03%).

Microwave‐Assisted Fabrication of Highly Crystalline, Robust COF Membrane for Organic Solvent Nanofiltration

AbstractFabrication of crystalline, robust covalent organic framework (COF) membranes based on disorder‐to‐order strategy is promising yet highly challenging. Herein, a microwave‐assisted method for fabricating COF membranes is proposed. Initially, monomers polymerize rapidly on the surface of porous Al2O3 substrate at room temperature to form an amorphous pristine membrane. Subsequently, a microwave field is exerted to trigger fast crystallization, acquiring a crystalline COF membrane within 60 min. The amorphous pristine membrane exhibits a high dissipation factor, indicating excellent microwave absorption capability, which accelerates the dynamic reversible reactions during the microwave treatment and thus ensures a rapid transition from the amorphous to the crystalline state. Owing to the high‐crystallinity and robust structure, the COF membranes exhibit high rejection rates for solute molecules with molecular weights exceeding 700 Da (e.g., Evans blue: 98.7%) and high solvent permeance for organic solvents (e.g., ethanol: 87.8 Lm−2h−1 bar−1, n‐hexane: 222.3 Lm−2h−1 bar−1). Surprisingly, the COF membranes exhibit superior mechanical properties, with Young's modulus of 33.91 ± 3.94 GPa, outperforming previously reported polycrystalline COF membranes and are close to those for inorganic zeolite membranes. The microwave‐assisted COF crystallization method opens a new avenue to fabricating a variety of crystalline membranes for advanced separations.

Effects of storage temperature on throat swabs preserved in different transport media for detection of SARS-CoV-2 by reverse transcriptase polymerase chain reaction

Objectives: The coronavirus disease 2019 (COVID-19) pandemic resulted in an increased need for molecular diagnostic testing. Delay in the specimen processing and storage of samples in laboratories leads to degradation of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) viral RNA. Inactivation transport medium (ITM) contains chaotropic agents that inactivate the virus and stabilize SARS-CoV-2 RNA for a longer duration, even at room temperature. The effect of different temperatures and duration of storage of samples in viral transport media (VTM) and ITM for detection of SARS-CoV-2 RNA was assessed. Materials and Methods: Samples from COVID-19 patients were aliquoted in ITM and VTM and kept at ambient temperature, 37°C and 45°C. SARS-CoV-2 viral RNA was extracted. Multiplex real-time polymerase chain reaction was done on days 0, 1, 3, and 5, and cycle threshold (Ct) values were noted. Statistical Analysis: Data were analyzed using the Statistical Package for the Social Sciences version 26.0. Linear variables were summarized as mean and standard deviations. One-way analysis of variance test with post hoc Tukey honestly significant difference was used to compare mean value between different loops and for pair-wise comparison. P < 0.05 was taken as significant. Results: The mean Ct values of both the Orf and E genes of the samples in VTM and ITM were stable across all temperature conditions on day 1. On day 5, the increase in Ct values for both E and Orf genes were significantly higher for VTM than ITM at ambient temperature, 37°C and 45°C. Ribonuclease P failure was significantly higher for VTM than ITM at ambient temperature and 37°C on day 3 and at all temperatures on day 5. Conclusions: ITM is a valuable transport media that can preserve SARS-CoV-2 for up to 5 days at ambient temperature and 37°C. As it renders the samples non-infectious, thus reducing the potential of biohazard events, this transport medium can be used effectively for the collection and transportation of SARS-CoV-2 samples, especially from remote or isolated healthcare facilities.

Debondable Epoxy-Acrylate Adhesives using β-Amino Ester Chemistry.

The reuse of multilayered materials, which are held together by structural epoxy adhesives, is a major challenge since the bonded substrates cannot be easily separated for recycling. In this research, we explore a one-pot strategy based on β-amino ester chemistry for the development of modified epoxy adhesives with on-demand debonding potential. For this, a formulation of commercially available acrylate, epoxy and amine compounds is used. The research starts with a systematic study, demonstrating the influence of the different compounds on the thermal and adhesive properties of the materials. Subsequently, the potential for debonding is demonstrated using rheological measurements and tensile tests. The fast, catalyst-free Aza-Michael reaction enables the straightforward preparation of such epoxy-based adhesives, while the reverse reaction allows for debonding at 120 °C. In general, a chemical design is demonstrated for producing an industrially attractive generation of debondable epoxy-based adhesives.

PNSC5325 prevents acute respiratory distress syndrome by alleviating inflammation and inhibiting extracellular matrix degradation of alveolar macrophages

BackgroundAcute respiratory distress syndrome (ARDS) is characterized by severe inflammation and significant extracellular matrix (ECM) degradation in the lungs. Our prior research identified the CtBP2-p300-NF-κB (C-terminal-binding protein 2-histone acetyltransferase p300-nuclear factor kappa B) transcriptional complex as critical in ARDS by activating pro-inflammatory cytokine genes. MethodsAn ARDS mouse model was established using intratracheal instillation of lipopolysaccharide (LPS). Small molecules that inhibit the CtBP2-p300 interaction were identified through AlphaScreen. RNA sequencing (RNA-Seq) was conducted to determine differential gene expression. Immunoprecipitation and co-immunoprecipitation analyzed protein interactions. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and immunoblotting detected gene and protein expression. Histological staining evaluated tissue damage. ResultsThrough AlphaScreen, two natural compounds, PNSC2477 and PNSC5325, were identified for their ability to inhibit the CtBP2-p300 interaction. While PNSC2477 demonstrated toxicity and was deemed unsuitable for further research, PNSC5325 exhibited minimal toxicity. PNSC5325 effectively inhibited the CtBP2-p300 interaction and reduced pro-inflammatory cytokine gene expression. RNA-Seq analysis of PNSC5325-treated cells indicated significant suppression of pro-inflammatory cytokine genes and matrix metalloproteinases (MMPs). Further molecular studies revealed that the CtBP2-p300 complex, in conjunction with activator protein 1 (AP1), activates MMP expression. PNSC5325 simultaneously suppressed both pro-inflammatory cytokines and MMPs by targeting the CtBP2-p300 complex. In LPS-injected mice, PNSC5325 administration significantly reduced ARDS incidence by inhibiting inflammatory and MMP genes. ConclusionThese findings suggest that PNSC5325 protects against ARDS by inhibiting key inflammatory and ECM degradation pathways, highlighting its potential as a novel therapeutic agent for ARDS and paving the way for further clinical investigations.

SNAP25 as a prognostic marker in transcriptome analysis of meningioma.

Meningiomas are the most common intracranial tumors and their diagnosis relies mostly on neuroimaging and histology. However, the histology grades cannot predict the outcome exactly and some meningiomas tend to recur after resection of even benign tumors. Therefore, it is necessary to explore prognostic and diagnostic molecular targets. Differential expression analysis between meningiomas and meninges was performed based on the merged data of GSE43290 and GSE84263. Next, we performed gene set enrichment analysis (GSEA), immune cell infiltration analysis, protein-protein interaction analysis, and survival analysis using public data. The expression level of Synaptosome-associated-protein-25kDa (SNAP25) was verified by reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) and Western blotting in meningioma tissues. There were 263 upregulated and 592 downregulated genes identified in meningiomas by differential expression analysis. GSEA results revealed that meningiomas were negatively related to the pathway of soluble N-ethylmaleimide sensitive factor attachment protein receptor interactions in vascular transport and chemokine signaling. SNAP25 was characterized as a hub gene and downregulated in meningiomas. The Kaplan-Meier plot indicated that high expression of SNAP25 is a favorable factor. SNAP25 was downregulated and identified as a potential prognostic marker in meningioma.

Sodium-Rich, Co-Ni-Free P2-Layered Manganese Oxide Cathodes for Sodium-Ion Batteries.

The rapid capacity loss attributed to irreversible phase reactions and structural instability has consistently affected the development of P2-layered cathode materials. Moreover, the introduction of costly elements such as single or multiple dopants has failed to resolve the sustainability challenges in designing an optimal Mn-based layered oxide cathode. This study proposes a Co-Ni-free, poly-elemental doping strategy (Li, Mg, and Cu) combined with high sodium content for an Mn-based P2-layered cathode designed for Na+ ion storage. In situ X-ray diffraction analysis confirms the absence of P2 - O2 phase transitions during cycling for both NLMMC87 (Na0.87Li0.1Mg0.1Mn0.7Cu0.1O2) and NLMMC77 (Na0.77Li0.1Mg0.1Mn0.7Cu0.1O2). This can be attributed to the co-substitution of the electronegative Cu element and the stabilizing dopants Li and Mg, which suppress oxygen evolution. Simultaneously, the high sodium content within the host structure promotes high reversible capacity, enhances structural stability, and minimizes internal stress due to volume changes. Moreover, NLMMC87 demonstrates a reversible capacity of 167.9mA h g-1 at 0.1 C (97% Coulombic efficiency) and maintains excellent stability across various current rates. Further investigations into the practical application of NLMMC87 in a sodium full cell will be a critical step toward realizing an ideal cathode for sodium-ion batteries.

Specific human gene expression in response to infection is an effective marker for diagnosis of latent and active tuberculosis

RNA sequencing and microarray analysis revealed transcriptional markers expressed in whole blood can differentiate active pulmonary TB (ATB) from other respiratory diseases (ORDs), and latent TB infection (LTBI) from healthy controls (HC). Here we describe a streamlined reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) assay that could be applied at near point-of-care for diagnosing and distinguishing ATB from ORDs and LTBI from HC. A literature review was undertaken to identify the most plausible host-gene markers (HGM) of TB infection. Primers, and dual labelled hydrolysis probes were designed and analytically evaluated for accuracy in an in-vitro model of infection using a lung fibroblast cell-line. Best performing genes were multiplexed into panels of 2–4 targets and taken forward for clinical evaluation. Mycobacteria Growth Indicator Tube and QuantiFERON-TB Gold Plus were used as reference tests for ATB and LTBI respectively. A total of 16 HGM were selected and incorporated into five multiplex RT-qPCR panels. PCR assay efficiency of all evaluated targets was ≥ 90% with a median analytical sensitivity of 292 copies/µl [IQR: 215.0-358.3 copies/µl], and a median limit of quantification of 61.7 copies/µl [IQR: 29.4-176.3 copies/µl]. Clinically, ATB was characterised by higher gene expression than ORDs, while LTBI was associated with lower gene expression than HC, Kruskal-Wallis p < 0.0001. Crucially, BATF2, CD64, GBP5, C1QB, GBP6, DUSP3, and GAS6 exhibited high differentiative ability for ATB from ORDs, LTBI or HC while KLF2, PTPRC, NEMF, ASUN, and ZNF296 independently discriminated LTBI from HC. Our results show that different HGM maybe required for ATB and LTBI differentiation from ORDs or HC respectively and demonstrate the feasibility of host gene-based RT-qPCR to diagnose ATB and LTBI at near point-of-care.