ABSTRACT
SERRATE (SE) is a core protein for microRNA (miRNA) biogenesis as well as for mRNA alternative splicing. Investigating the regulatory mechanism of SE expression is hence critical to understanding its detailed function in diverse biological processes. However, little about the control of SE expression has been clarified, especially through long noncoding RNA (lncRNA). Here, we identified an antisense intragenic lncRNA transcribed from the 3' end of SE, named SEAIRa. SEAIRa repressed SE expression, which in turn led to serrated leaves. SEAIRa recruited plant U-box proteins PUB25/26 with unreported RNA binding ability and a ubiquitin-like protein related to ubiquitin 1 (RUB1) for H2A monoubiquitination (H2Aub) at exon 11 of SE. In addition, PUB25/26 helped cleave SEAIRa and release the 5' domain fragment, which recruited the PRC2 complex for H3 lysine 27 trimethylation (H3K27me3) deposition at the first exon of SE. The distinct modifications of H2Aub and H3K27me3 at different sites of the SE locus cooperatively suppressed SE expression. Collectively, our results uncover an epigenetic mechanism mediated by the lncRNA SEAIRa that modulates SE expression, which is indispensable for plant growth and development.
Subject(s)
Arabidopsis Proteins , Arabidopsis , Epigenetic Repression , RNA, Long Noncoding , RNA-Binding Proteins , Epigenesis, Genetic , Histones , RNA, Long Noncoding/genetics , Arabidopsis Proteins/genetics , RNA-Binding Proteins/geneticsABSTRACT
COLD is a major naturally occurring stress that usually causes complex symptoms and severe yield loss in crops. R-loops function in various cellular processes, including development and stress responses, in plants. However, how R-loops function in COLD responses is largely unknown in COLD susceptible crops like rice (Oryza sativa L.). We conducted DRIP-Seq along with other omics data (RNA-Seq, DNase-Seq and ChIP-Seq) in rice with or without COLD treatment. COLD treatment caused R-loop reprogramming across the genome. COLD-biased R-loops had higher GC content and novel motifs for the binding of distinct transcription factors (TFs). Moreover, R-loops can directly/indirectly modulate the transcription of a subset of COLD-responsive genes, which can be mediated by R-loop overlapping TF-centered or cis-regulatory element-related regulatory networks and lncRNAs, accounting for c. 60% of COLD-induced expression of differential genes in rice, which is different from the findings in Arabidopsis. We validated two R-loop loci with contrasting (negative/positive) roles in the regulation of two individual COLD-responsive gene expression, as potential targets for enhanced COLD resistance. Our study provides detailed evidence showing functions of R-loop reprogramming during COLD responses and provides some potential R-loop loci for genetic and epigenetic manipulation toward breeding of rice varieties with enhanced COLD tolerance.
Subject(s)
Arabidopsis , Oryza , Oryza/metabolism , R-Loop Structures , Plant Proteins/metabolism , Plant Breeding , Transcription Factors/metabolism , Arabidopsis/genetics , Gene Expression Regulation, Plant , Cold TemperatureABSTRACT
Cis-regulatory elements (CREs) fine-tune gene transcription in eukaryotes. CREs with sequence variations play vital roles in driving plant or crop domestication. However, how global sequence and structural variations (SVs) are responsible for multilevel changes between indica and japonica rice (Oryza sativa) is still not fully elucidated. To address this, we conducted multiomic studies using MNase hypersensitivity sequencing (MH-seq) in combination with RNA sequencing (RNA-seq), chromatin immunoprecipitation sequencing (ChIP-seq), and bisulfite sequencing (BS-seq) between the japonica rice variety Nipponbare (NIP) and indica rice variety 93-11. We found that differential MNase hypersensitive sites (MHSs) exhibited some distinct intrinsic genomic sequence features between NIP and 93-11. Notably, through MHS-genome-wide association studies (GWAS) integration, we found that key sequence variations may be associated with differences of agronomic traits between NIP and 93-11, which is partly achieved by MHSs harboring CREs. In addition, SV-derived differential MHSs caused by transposable element (TE) insertion, especially by noncommon TEs among rice varieties, were associated with genes with distinct functions, indicating that TE-driven gene neo- or subfunctionalization is mediated by changes of chromatin openness. This study thus provides insights into how sequence and genomic SVs control agronomic traits of NIP and 93-11; it also provides genome-editing targets for molecular breeding aiming at improving favorable agronomic properties.
Subject(s)
Oryza , Oryza/genetics , Chromatin/genetics , Genome-Wide Association Study , Epigenesis, Genetic , DNA Transposable Elements/genetics , Genomics , Genome, Plant/geneticsABSTRACT
I-motifs (iMs) are non-canonical DNA secondary structures that fold from cytosine (C)-rich genomic DNA regions termed putative i-motif forming sequences (PiMFSs). The structure of iMs is stabilized by hemiprotonated C-C base pairs, and their functions are now suspected in key cellular processes in human cells such as genome stability and regulation of gene transcription. In plants, their biological relevance is still largely unknown. Here, we characterized PiMFSs with high potential for i-motif formation in the rice genome by developing and applying a protocol hinging on an iMab antibody-based immunoprecipitation (IP) coupled with high-throughput sequencing (seq), consequently termed iM-IP-seq. We found that PiMFSs had intrinsic subgenomic distributions, cis-regulatory functions and an intricate relationship with DNA methylation. We indeed found that the coordination of PiMFSs with DNA methylation may affect dynamics of transposable elements (TEs) among different cultivated Oryza subpopulations or during evolution of wild rice species. Collectively, our study provides first and unique insights into the biology of iMs in plants, with potential applications in plant biotechnology for improving important agronomic rice traits.
Subject(s)
DNA Transposable Elements , Oryza , Cytosine , DNA Methylation , DNA Transposable Elements/genetics , Genome, Plant , High-Throughput Nucleotide Sequencing , Humans , Oryza/geneticsABSTRACT
Rapid radiative transfer models are crucial to remote sensing and data assimilation. An integrated efficient radiative transfer model named Dayu, which is an updated version of the Efficient Radiative Transfer Model (ERTM) is developed to simulate the imager measurements in cloudy atmospheres. In Dayu model, the Optimized alternate Mapping Correlated K-Distribution model (OMCKD) which is predominant in dealing with the overlap of multiple gaseous lines is employed to efficiently calculate the gaseous absorption. The cloud and aerosol optical properties are pre-calculated and parameterized by the particle effective radius or length. Specifically, the ice crystal model is assumed as a solid hexagonal column, of which the parameters are determined based on massive aircraft observations. For the radiative transfer solver, the original 4-stream Discrete ordinate aDding Approximation (4-DDA) is extended to 2N-DDA (2N is the number of streams) which can calculate not only the azimuthally dependent radiance in the solar spectrum (including solar and infrared spectra overlap) but also the azimuthally averaged radiance in the thermal infrared spectrum through a unified adding method. Then the accuracy and efficiency of Dayu model are evaluated by comparing it with the benchmark model, i.e., Line-By-Line Radiative Transfer Model (LBLRTM) and DIScrete Ordinate Radiative Transfer (DISORT). Under the standard atmospheric profile, the maximum relative biases between Dayu model with 8-DDA / 16-DDA and the benchmark model (OMCKD with 64-stream DISORT) are 7.63% / 2.62% at solar channels but decreased to 2.66% / 1.39% at spectra-overlapping channel (3.7 µm). The computational efficiency of Dayu model with 8-DDA / 16-DDA is approximately three / two orders of magnitude higher than that of the benchmark model. At thermal infrared channels, the brightness temperature (BT) differences between Dayu model with 4-DDA and the benchmark model (LBLRTM with 64-stream DISORT) are bounded by 0.65K. Compared to the benchmark model, Dayu model with 4-DDA improves the computational efficiency by five orders of magnitude. In the application to the practical Typhoon Lekima case, the simulated reflectances and BTs by Dayu model have a high consistency with the imager measurements, demonstrating the superior performance of Dayu model in satellite simulation.
ABSTRACT
BACKGROUND: The United Nations calculates there were 703 million adults 65 years and older globally as of 2019 with this number projected to double by 2050. A significant number of older adults live with comorbid health conditions, making the role of a nurse in long-term care (LTC) complex. Our objective was to identify the challenges, facilitators, workload, professional development and clinical environment issues that influence nurses and nursing students to seek work and continue to work in LTC settings. METHODS: Eligibility criteria included being a nurse in a LTC setting and research with a substantial qualitative component. Multiple databases (including Medline and CINAHL) were searched between 2013 and 2019 along with grey literature. Covidence was used to organise a team of 10 into a paired review of titles and abstracts to the final full text screening, extraction and appraisal with the CASP Qualitative Studies Checklist. Analysis involved a thematic synthesis approach. The Enhancing Transparency in Reporting the Synthesis of Qualitative Research (ENTREQ) checklist informed the writing of the review. RESULTS: The search resulted in 18 articles and dissertations. Areas investigated included recruitment, resilience, employment and retention, how nurses perceived their professional work, rewards and difficulties, supervision, student preceptorship and career aspiration, nurses' perceptions of occupational status, along with leadership, education and development needs, and intentions to manage resident deteriorating health. The five themes were (1) perspectives of nursing influenced by the organisation, (2) pride in, and capacity to build relationships, (3) stretching beyond the technical skills, (4) autonomy, and (5) taking on the challenge of societal perceptions. DISCUSSION: This review revealed what is required to recruit nursing students to careers in LTC and retain nurses. To be explored is how staff can work to their full scope of practice and the resultant impact on resident care, including how to maximise a meaningful life for residents and their families. REGISTRATION: National Institute for Health Research UK (Prospero ID: CRD42019125214).
Subject(s)
Long-Term Care , Nurses , Humans , Aged , Students , Employment , Qualitative ResearchABSTRACT
Satellite data assimilation requires a computationally fast and accurate radiative transfer model. Currently, three fast models are commonly used in the Numerical Weather Prediction models (NWP) for satellite data assimilation, including Radiative Transfer for TIROS Operational Vertical Sounder (RTTOV), Community Radiative Transfer Model (CRTM), and Advanced Radiative transfer Modeling System (ARMS). ARMS was initiated in 2018 and is now becoming the third pillar supporting many users in NWP and remote sensing fields. Its radiative transfer solvers (e.g. Doubling Adding method) is inherited from CRTM. In this study, we propose a Discrete Ordinate Adding Method (DOAM) to solve the radiative transfer equation including both solar and thermal source terms. In order to accelerate the DOAM computation, the single scattering approximation is used in the layer with an optical depth less than 10-8 or a single scattering albedo less than 10-10. From principles of invariance, the adding method is then applied to link the radiances between the layers. The accuracy of DOAM is evaluated through four benchmark cases. It is shown that the difference between DOAM and DIScrete Ordinate Radiative Transfer (DISORT) decreases with an increase of stream number. The relative bias of the 4-stream DOAM ranges from -5.03 % to 5.92 % in the triple layers of a visible wavelength case, while the maximum bias of the 8-stream DOAM is only about 1 %. The biases can be significantly reduced by the single scattering correction. Comparing to the visible case, the accuracy of the 4-stream DOAM is much higher in the thermal case with a maximum bias -1.69 %. Similar results are also shown in two multiple-layer cases. In the MacBook Pro (15-inch, 2018) laptop, the 2-stream DOAM only takes 1.68 seconds for calculating azimuthally independent radiance of 3000 profiles in the hyper-spectral oxygen A-band (wavelength ranges from 0.757 µm to 0.775 µm), while the 4-stream DOAM takes 4.06 seconds and the 16-stream DOAM takes 45.93 seconds. The time of the 2-, 4- and 16- stream DOAM are 0.86 seconds, 1.09 seconds and 4.34 seconds for calculating azimuthally averaged radiance. DISORT with 16 streams takes 1521.56 seconds and 127.64 seconds under the same condition. As a new solver, DOAM has been integrated into ARMS and is used to simulate the brightness temperatures at MicroWave Humidity Sounder (MWHS) as well as MicroWave Radiation Imager (MWRI) frequencies. The simulations by DOAM are compared to those by Doubling Adding method and accuracy of both solvers shows a general agreement. All the results show that the DOAM is accurate and computational efficient for applications in NWP data assimilation and satellite remote sensing.
ABSTRACT
BACKGROUND: This study aimed to explore the effects of ceramide (Cer) on NLRP3 inflammasome activation and their underlying mechanisms. METHODS: Lipopolysaccharide (LPS)/adenosine triphosphate (ATP)-induced NLRP3 inflammasome activation in J774A.1 cells and THP-1 macrophages was used as an in vitro model of inflammation. Western blotting and real-time PCR (RT-PCR) were used to detect the protein and mRNA levels, respectively. IL-1ß and IL-18 levels were measured by ELISA. ASM assay kit and immunofluorescence were used to detect ASM activity and Cer content. RESULTS: Imipramine, a well-known inhibitor of ASM, significantly inhibited LPS/ATP-induced activity of ASM and the consequent accumulation of Cer. Additionally, imipramine suppressed the LPS/ATP-induced expression of thioredoxin interacting protein (TXNIP), NLRP3, caspase-1, IL-1ß, and IL-18 at the protein and mRNA level. Interestingly verapamil, a TXNIP inhibitor, suppressed LPS/ATP-induced activation of TXNIP/NLRP3 inflammasome but did not affect LPS/ATP-induced ASM activation and Cer formation. TXNIP siRNA and verapamil inhibited C2-Cer-induced upregulation of TXNIP and activation of the NLRP3 inflammasome. In addition, the pretreatment of cells with sulfo-N-succinimidyl oleate (SSO), an irreversible inhibitor of the scavenger receptor CD36, blocked Cer-induced upregulation of nuclear factor-κB (NF-κB) activity, TXNIP expression, and NLRP3 inflammasome activation. Inhibition of NF-κB activation by SN50 prevented Cer-induced upregulation of TXNIP and activation of the NLRP3 inflammasome but did not affect CD36 expression. CONCLUSION: This study demonstrated that the ASM/Cer/TXNIP signaling pathway is involved in NLRP3 inflammasome activation. The results documented that the CD36-dependent NF-κB-TXNIP signaling pathway plays an essential role in the Cer-induced activation of NLRP3 inflammasomes in macrophages.
Subject(s)
Carrier Proteins/genetics , Ceramides/genetics , Inflammation/genetics , NLR Family, Pyrin Domain-Containing 3 Protein/genetics , Adenosine Triphosphate/metabolism , Caspase 1/genetics , Gene Expression Regulation/genetics , Humans , Inflammasomes/genetics , Inflammation/metabolism , Inflammation/pathology , Interleukin-18/genetics , Interleukin-1beta/genetics , Lipopolysaccharides/pharmacology , Macrophages/drug effects , NF-kappa B/antagonists & inhibitors , NF-kappa B/genetics , Oleic Acids/pharmacology , Peptides/pharmacology , Signal Transduction/drug effects , Sphingomyelin Phosphodiesterase/genetics , Succinimides/pharmacology , Verapamil/pharmacologyABSTRACT
R-loops and covalent modifications of N 6 -methyladenine on DNA (D-6 mA) or RNA (R-m6A) have been documented to function in various cellular processes in eukaryotes. However, the relationships between R-loops and both covalent modifications are still elusive in plants. Here, we integrated existing ssDRIP-seq with D-6 mA and R-m6A data from Arabidopsis thaliana. We found that the presence of either of both modifications facilitates R-loop formation and transcription of overlapping genes. Interestingly, our study suggests that the presence of R-m6A is key to affect R-loop intensity and positively regulate gene transcription. Moreover, the presence of D-6 mA plays an additive role to facilitate the effect of R-m6A on R-loop intensity, however, D-6 mA may negatively regulate gene transcription when coexisted with R-m6A. Our analyses indicate that D-6 mA, R-m6A, or histone marks may act individually and cooperatively with R-loops in regulating gene transcription. Our study is the first to link R-loops with D-6 mA and R-m6A in plants, thereby providing new insights into interactions between R-loops with D-6 mA, R-m6A, and histone marks for regulating gene transcription. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01010-5.
ABSTRACT
An efficient radiative transfer model (ERTM) is developed to simulate thermal infrared brightness temperatures observed by the Advanced Himawari Imager (AHI) in this study. The ERTM contains an alternate mapping correlated k-distribution (AMCKD) scheme, a parameterization for cloud optical property, and a rapid infrared radiative transfer scheme. The AMCKD is employed to calculate the gaseous absorption in the inhomogeneous thermodynamic atmosphere. The optical properties of clouds are parameterized by the effective length for ice clouds based on the Voronoi model, and by the effective radius for water clouds based on the Lorenz-Mie theory. The adding method of four-stream discrete ordinates method (4DDA) is extended to be able to calculate the thermal infrared radiative intensity varying with the zenith angle in ERTM. The efficiency and accuracy of ERTM are evaluated by comparing with the benchmark model which is composed of discrete ordinate radiative transfer (DISORT) and line-by-line radiative transfer model (LBLRTM). Under the standard atmospheric profiles, the root mean square error (RMSE) of simulated brightness temperatures reaches a maximum of 0.21K at the B16 (13.28 µm) channel of AHI. The computational efficiency of ERTM is approximately five orders of magnitude higher than that of the benchmark model. Moreover, the simulated brightness temperatures by ERTM are highly consistent with the rigorous results and AHI observations in the application to the Typhoon Mujigae case.
ABSTRACT
Salvianolic acid B (Sal B) exerts strong antioxidant activity and eliminates the free radical effect. However, how it affects the antioxidant pathway is not very clear. The objective of this study was to investigate the underlying mechanism of Sal B in CCl4-induced acute liver injury, especially its effect on the Nrf2/HO-1 signaling pathway. For the in vivo experiment, an acute liver injury model was induced using CCl4 and treated with Sal B. For the in vitro experiment, an oxidative damage model was established followed by Sal B treatment. Serum biochemical indicators and reactive oxygen species activity were detected using corresponding kits. Oxidant/antioxidant status was determined based on the levels of malondialdehyde, glutathione, and superoxide dismutase. Nrf2 and HO-1 levels were analyzed by Western blotting and immunohistochemical staining. Sal B treatment improved liver histology, decreased the aminotransferase levels, and attenuated oxidative stress in the acute liver injury model. Nrf2 and HO-1 levels were increased both in vivo and in vitro. Sal B suppresses acute liver injury and Nrf2/HO-1 signaling plays a key role in this process.
Subject(s)
Benzofurans/pharmacology , Carbon Tetrachloride/toxicity , Chemical and Drug Induced Liver Injury/prevention & control , Gene Expression Regulation/drug effects , Heme Oxygenase-1/metabolism , Membrane Proteins/metabolism , NF-E2-Related Factor 2/metabolism , Animals , Apoptosis , Chemical and Drug Induced Liver Injury/etiology , Chemical and Drug Induced Liver Injury/metabolism , Chemical and Drug Induced Liver Injury/pathology , Glutathione/metabolism , Heme Oxygenase-1/genetics , Male , Membrane Proteins/genetics , Mice , NF-E2-Related Factor 2/genetics , Oxidative Stress/drug effects , Reactive Oxygen Species/metabolism , Signal TransductionABSTRACT
A multi-layer solar radiative transfer (RT) scheme is proposed to deal with the vertical variation of inherent microphysical properties of clouds in this study. The exponential expressions are used to represent the vertical variation of optical properties caused by inhomogeneous microphysical properties. A perturbation method, coupled with the Eddington approximation, is used to solve the RT equation. In order to have a more accurate estimation of reflectance/transmittance for every single layer, the optical properties are adjusted following the theory of delta scaling in the proposed scheme. In addition, a modified adding method based on Chandrasekhar's invariance principle is introduced to solve the multi-layer RT. The accuracy of the proposed scheme is investigated by comparing the reflectance/absorptance to the benchmark for two double-layer cases, and each layer with vertically inhomogeneous optical properties. Results show that the bias related to vertically inhomogeneous optical properties reaches 13.8 % for reflectance and 29.2 % for absorptance while the bias of the proposed scheme is only -0.8 % for reflectance and 1.7 % for absorptance. We also apply the proposed scheme as well as the conventional Eddington approximation to the CanadianClimate Center RT model which handle RT in CanAM4. The calculations are performed in the following four solar wavenumber bands 2500-4200, 4200-8400, 8400-14500 and 14500-50000 cm -1. The result also shows that the proposed scheme also improved the accuracy in both flux and heating rate calculation by taking the vertical variation of inherent microphysical properties into account. The proposed scheme is approximately three times more computationally expensive compared to the Eddington approximation when we only consider the algorithm itself. The computational time is doubled compared to the Eddington approximation when we take the complete radiative transfer process into account. Due to its accuracy and efficiency, the proposed scheme is suitable to improve the RT calculations regarding the vertical variation of inherent microphysical properties in climate models.
ABSTRACT
KEY MESSAGE: We report coding and long noncoding RNAs in maize upon phytohormone gibberellin stimulation. Plant hormone gibberellin (GA) orchestrates various facets of biological processes. Dissection the transcriptomic dynamics upon GA stimulation has biological significance. Feature of maize transcriptome in response to GA application remains largely elusive. Herein, two types of plants, one was with normal height, the other was GA-sensitive dwarfism, were selected from advanced backcross population for GA3 treatment with different concentrations. In control and GA3-treated plants, we identified a large number of coding and long noncoding RNAs (lncRNAs) through sequencing eight ribosomal-depleted RNA libraries. Transcripts encoding GA biosynthetic and metabolic enzymes KS, GA20ox, GA3ox, and GA2ox were significantly differentially expressed in GA3-treated samples. A total of 78 protein-coding transcripts were shared between GA3-treated normal height and dwarf plants. Shared transcripts encoding terpene synthase, MYB transcription factor, and receptor-like protein kinase were co-regulated with their corresponding partners. Out of identified lncRNAs, 22 and 34 significantly differentially expressed lncRNAs were responsive to GA application in normal height and dwarf plants, respectively. Shared GA-responsive lncRNAs were found in GA3-treated normal height and dwarf plants. Some lncRNAs corresponded to precursors of known miRNA, such as zma-miR528a and zma-miR528b. Multiple promising targets of significantly differentially expressed lncRNAs were discovered, including Lazy plant1 for auxin- and GA-mediated shoot gravitropism, bZIP transcription factor member for GA-controlled cell elongation. This study will improve our knowledge of GA-triggered transcriptome change and facilitate a comprehensive understanding of regulatory cascade centering on GA.
Subject(s)
Gibberellins/pharmacology , Plant Growth Regulators/pharmacology , RNA, Long Noncoding/drug effects , Zea mays/genetics , Gene Expression Regulation, Plant/drug effects , Gene Expression Regulation, Plant/genetics , Gene Ontology , RNA, Long Noncoding/genetics , RNA, Long Noncoding/metabolism , Real-Time Polymerase Chain Reaction , Sequence Analysis, RNA , Zea mays/drug effects , Zea mays/metabolismABSTRACT
This is the protocol for a Cochrane Review. The objectives are as follows: The aim of this systematic review is to advance our understanding of the key characteristics of effective preschool-based interventions designed to foster self-regulation. To accomplish this, the review addresses the following questions: 1. What types of preschool-based interventions have been developed to promote self-regulation? 2. What is the average effect of these preschool-based interventions on self-regulation, focusing on four key constructs: integrative effortful control, integrative executive function, self-regulation, and self-regulated learning? 3. What characteristics-such as Resource Allocation, Activity Type, and Instruction Method-could potentially contribute to the effects of preschool-based interventions in promoting self-regulation?
ABSTRACT
Trastuzumab is effective in the treatment of HER2/neu over-expressing breast cancer, but not all patients benefit from it. In vitro data suggest a role for HER3 in the initiation of signaling activity involving the AKTmTOR pathway leading to trastuzumab insensitivity. We sought to investigate the potential of HER3 alone and in the context of p95HER2 (p95), a trastuzumab resistance marker, as biomarkers of trastuzumab escape. Using the VeraTag® assay platform, we developed a dual antibody proximity-based assay for the precise quantitation of HER3 total protein (H3T) from formalin-fixed paraffin-embedded (FFPE) breast tumors. We then measured H3T in 89 patients with metastatic breast cancer treated with trastuzumab-based therapy, and correlated the results with progression-free survival and overall survival using KaplanMeier and decision tree analyses that also included HER2 total (H2T) and p95 expression levels. Within the sub-population of patients that over-expressed HER2, high levels of HER3 and/or p95 protein expression were significantly associated with poor clinical outcomes on trastuzumab-based therapy. Based on quantitative H3T, p95, and H2T measurements, multiple subtypes of HER2-positive breast cancer were identified that differ in their outcome following trastuzumab therapy. These data suggest that HER3 and p95 are informative biomarkers of clinical outcomes on trastuzumab therapy, and that multiple subtypes of HER2-positive breast cancer may be defined by quantitative measurements of H3T, p95, and H2T.
Subject(s)
Biomarkers, Tumor/analysis , Breast Neoplasms/secondary , Fluorescent Antibody Technique, Indirect , Gene Expression Regulation, Neoplastic , Genes, erbB-2 , Neoplasm Proteins/biosynthesis , Receptor, ErbB-2/analysis , Receptor, ErbB-3/analysis , Antibodies, Monoclonal/immunology , Antibodies, Monoclonal, Humanized/pharmacology , Antibodies, Monoclonal, Humanized/therapeutic use , Antineoplastic Agents/pharmacology , Antineoplastic Agents/therapeutic use , Biomarkers, Tumor/genetics , Biomarkers, Tumor/immunology , Breast Neoplasms/classification , Breast Neoplasms/drug therapy , Breast Neoplasms/genetics , Breast Neoplasms/metabolism , Breast Neoplasms/mortality , Cell Line, Tumor , Cohort Studies , Decision Trees , Disease-Free Survival , Drug Resistance, Neoplasm , Female , Humans , Kaplan-Meier Estimate , Neoplasm Proteins/genetics , Peptide Fragments/analysis , Peptide Fragments/immunology , Prognosis , Protein Structure, Tertiary , Receptor, ErbB-2/genetics , Receptor, ErbB-2/immunology , Receptor, ErbB-3/genetics , Receptor, ErbB-3/immunology , Retrospective Studies , Single-Blind Method , Trastuzumab , Treatment OutcomeABSTRACT
A hierarchical S/MWCNT nanomicrosphere for lithium/sulfur batteries with a high power and energy density as well as an excellent cycle life is introduced. Sulfur was uniformly coated on the surface of functional MWCNTs, which serves as a carbon matrix, to form a typical nanoscale core-shell structure with a sulfur layer of thickness 10-20 nm. Then the nanoscale sulfur intermediate composite was ball-milled to form interwoven and porous sphere architecture with large pores (around 1 µm to 5 µm). Different from most sulfur/carbon materials with micropore and mesopore structure, the micrometre scale S/MWCNT nanomicrosphere with a large pore structure could also exhibit high sulfur utilization and cycle retention. It could maintain a reversible capacity of 1000 mA h g(-1) after 100 cycles at 0.3 A g(-1) current density. And it even remained 780 mA h g(-1) after 200 cycles at 0.5 A g(-1) and 650 mA h g(-1) after 200 cycles at 1 A g(-1), showing a significant cyclability enhancement. It is believed that under the collective effect of hierarchical architecture, as well as the existence of carboxyl functional groups, sulfur/carbon materials with large pores could also exhibit an excellent electrochemical performance. The synthesis process introduced here is simple and broadly applicable, which would not only be beneficial to design new materials for lithium sulfur batteries but can also be extended to many different electrode materials for lithium ion batteries.
Subject(s)
Electric Power Supplies , Lithium/chemistry , Nanotubes, Carbon/chemistry , Sulfur/chemistry , MicrospheresABSTRACT
BACKGROUND: Pyroptosis of endothelial cells is a new cause of endothelial dysfunction in multiple diseases. Ceramide acts as a potential bioactive mediator of inflammation and increases vascular endothelial permeability in many diseases, whether it can aggravate vascular endothelial injury by inducing cell pyroptosis remains unknown. This study was established to explore the effects of C8-ceramide (C8-Cer) on human umbilical vein vascular endothelial cells (HUVECs) and its possible underlying mechanism. METHODS: HUVECs were exposed to various concentrations of C8-Cer for 12 h, 24 h, 48 h. The cell survival rate was measured using the cell counting kit-8 assay. Western blotting and Real-time polymerase chain reaction (RT-PCR) were used to detect the pyroptosis-releated protein and mRNA expressions, respectively. Caspase-1 activity assay was used to detect caspase-1 activity. Hoechst 33342/propidium iodide double staining and flow cytometry were adopted to measure positive staining of cells. Lactate dehydrogenase release assay and enzyme-linked immunosorbent assay were adopted to measure leakage of cellular contents. FITC method was used to detect the permeability of endothelial cells. ROS fluorescence intensity were detected by flow cytometry. RESULTS: The viability of HUVECs decreased gradually with the increase in ceramide concentration and time. Ceramide upregulated the expression of thioredoxin interacting protein (TXNIP), NLRP3, GSDMD, GSDMD-NT, caspase-1 and Casp1 p20 at the protein and mRNA level in a dose-dependent manner. It also enhanced the PI uptake in HUVECs and upregulated caspase-1 activity. Moreover, it promoted the release of lactate dehydrogenase, interleukin-1ß, and interleukin-18. Meanwhile, we found that ceramide led to increased vascular permeability. The inhibitor of NLRP3 inflammasome assembly, MCC950, was able to disrupt the aforementioned positive loop, thus alleviating vascular endothelial cell damage. Interestingly, inhibition of TXNIP either chemically using verapamil or genetically using small interfering RNA (siRNA) can effectively inhibit ceramide-induced pyroptosis and improved cell permeability. In addition, ceramide stimulated reactive oxygen species (ROS) generation. The pretreatment of antioxidant N-acetylcysteine (NAC), ROS scavenger, blocked the expression of pyroptosis markers induced by C8-cer in HUVECs. CONCLUSION: The current study demonstrated that C8-Cer could aggravate vascular endothelial cell damage and increased cell permeability by inducing cell pyroptosis. The results documented that the ROS-dependent TXNIP/NLRP3/GSDMD signalling pathway plays an essential role in the ceramide-induced pyroptosis in HUVECs.
Subject(s)
NLR Family, Pyrin Domain-Containing 3 Protein , Pyroptosis , Humans , Pyroptosis/genetics , NLR Family, Pyrin Domain-Containing 3 Protein/genetics , NLR Family, Pyrin Domain-Containing 3 Protein/metabolism , Reactive Oxygen Species/metabolism , Ceramides/pharmacology , Caspase 1/genetics , Caspase 1/metabolism , Caspase 1/pharmacology , Human Umbilical Vein Endothelial Cells , RNA, Messenger/genetics , Lactate Dehydrogenases/metabolism , Carrier Proteins , Phosphate-Binding Proteins/metabolism , Phosphate-Binding Proteins/pharmacology , Pore Forming Cytotoxic Proteins/metabolismABSTRACT
BACKGROUND: Maize mesophyll (M) cells play important roles in various biological processes such as photosynthesis II and secondary metabolism. Functional differentiation occurs during M-cell development, but the underlying mechanisms for regulating M-cell development are largely unknown. RESULTS: We conducted single-cell RNA sequencing (scRNA-seq) to profile transcripts in maize leaves. We then identified coregulated modules by analyzing the resulting pseudo-time-series data through gene regulatory network analyses. WRKY, ERF, NAC, MYB and Heat stress transcription factor (HSF) families were highly expressed in the early stage, whereas CONSTANS (CO)-like (COL) and ERF families were highly expressed in the late stage of M-cell development. Construction of regulatory networks revealed that these transcript factor (TF) families, especially HSF and COL, were the major players in the early and later stages of M-cell development, respectively. Integration of scRNA expression matrix with TF ChIP-seq and Hi-C further revealed regulatory interactions between these TFs and their targets. HSF1 and COL8 were primarily expressed in the leaf bases and tips, respectively, and their targets were validated with protoplast-based ChIP-qPCR, with the binding sites of HSF1 being experimentally confirmed. CONCLUSIONS: Our study provides evidence that several TF families, with the involvement of epigenetic regulation, play vital roles in the regulation of M-cell development in maize.
Subject(s)
Transcription Factors , Zea mays , Epigenesis, Genetic , Gene Expression Regulation, Plant , Humans , Mesophyll Cells/metabolism , Transcription Factors/metabolism , Transcriptome/genetics , Zea mays/genetics , Zea mays/metabolismABSTRACT
Ceramide is a biologically active sphingomyelin that inhibits cell growth and proliferation. In previous studies, it was demonstrated that the use of lipopolysaccharides induces acid sphingomyelinases to produce ceramide, promoting lung cancer cell apoptosis; however, the specific mechanisms of this action remain unclear. Thioredoxininteracting protein (Txnip) plays an important role in the signal transmission of redox reactions inside and outside the cell. Thus, it was hypothesized that ceramide induces apoptosis in lung adenocarcinoma cells (A549 and PC9) by modulating the Txnip/Trx1 complex. In the present study, the Cell Counting kit8 method was used to detect cell activity and the drug concentration. Hoechst 33258 staining and flow cytometry were used to detect cell apoptosis, and the positional association between Txnip and Trx1 upregulated by ceramide was observed by immunofluorescence confocal microscopy. Reverse transcriptionquantitative polymerase chain reaction and western blot analysis were used to detect the changes in related gene, mRNA and protein expression levels. The results revealed that ceramide treatment resulted in the upregulation of Txnip and in the reduction of Trx1 activities. However, the Txnip inhibitor, verapamil, reversed these changes. The analysis of mRNA expression further verified the changes observed in the protein expression of Txnip, Trx1 and apoptosisrelated proteins. On the whole, the present study demonstrates that ceramide induces the apoptosis of lung cancer cells by regulating the Txnip/Trx1 complex.