Pre-mRNA splicing repression triggers abiotic stress signaling in plants

[EN] Alternative splicing (AS) of precursor RNAs enhances transcriptome plasticity and proteome diversity in response to diverse growth and stress cues. Recent work has shown that AS is pervasive across plant species, with more than 60% of intron-containing genes producing different isoforms. Mammalian cell-based assays have discovered various inhibitors of AS. Here, we show that the macrolide pladienolide B (PB) inhibits constitutive splicing and AS in plants. Also, our RNA sequencing (RNA-seq) data revealed that PB mimics abiotic stress signals including salt, drought and abscisic acid (ABA). PB activates the abiotic stress-and ABA-responsive reporters RD29A::LUC and MAPKKK18::uidA in Arabidopsis thaliana and mimics the effects of ABA on stomatal aperture. Genome-wide analysis of AS by RNA-seq revealed that PB perturbs the splicing machinery and leads to a striking increase in intron retention and a reduction in other forms of AS. Interestingly, PB treatment activates the ABA signaling pathway by inhibiting the splicing of clade A PP2C phosphatases while still maintaining to some extent the splicing of ABA-activated SnRK2 kinases. Taken together, our data establish PB as an inhibitor and modulator of splicing and a mimic of abiotic stress signals in plants. Thus, PB reveals the molecular underpinnings of the interplay between stress responses, ABA signaling and post-transcriptional regulation in plants.We wish to thank members of the Laboratory for Genome Engineering at King Abdullah University of Science and Technology for helpful discussions and comments on the manuscript. We wish to thank Moussa Benhamed for helpful discussions and suggestions and for providing key materials. We wish to thank Sean Cutler for providing Arabidopsis seeds of MAKPKKK18-uidA. This study was supported by King Abdullah University of Science and Technology. Work in PR's laboratory was funded by grant BIO2014-52537-R from MINECO. Work in PD's laboratory is funded by grant PTDC/BIA-PLA/1084/2014 from FCT. The authors declare no conflicts of interest.Ling, Y.; Alshareef, S.; Butt, H.; Lozano Juste, J.; Li, L.; Galal, AA.; Moustafa, A.... (2017). Pre-mRNA splicing repression triggers abiotic stress signaling in plants. The Plant Journal. 89(2):291-309. https://doi.org/10.1111/tpj.13383S29130989

Pre-mRNA splicing repression triggers abiotic stress signaling in plants

Alternative splicing (AS) of precursor RNAs enhances transcriptome plasticity and proteome diversity in response to diverse growth and stress cues. Recent work showed that AS is pervasive across plant species, with more than 60% of intron-containing genes producing different isoforms. Mammalian cell-based assays have discovered various AS inhibitors. Here, we show that the macrolide Pladienolide B (PB) inhibits constitutive splicing and AS in plants. Also, our RNA-seq data revealed that PB mimics abiotic stress signals including salt, drought, and abscisic acid (ABA). PB activates the abiotic stress- and ABA-responsive reporters RD29A::LUC and MAPKKK18::GUS in Arabidopsis thaliana and mimics the effects of ABA on stomatal aperture. Genome-wide analysis of AS by RNA-seq revealed that PB perturbs the splicing machinery and leads to a striking increase in intron retention and a reduction in other forms of AS. Interestingly, PB treatment activates the ABA signaling pathway by inhibiting the splicing of clade A PP2Cs phosphatases while still maintaining to some extent the splicing of ABA-activated SnRK2 kinases. Taken together, our data establish PB as an inhibitor and modulator of splicing and a mimic of abiotic stress signals in plants. Thus, PB reveals the molecular underpinnings of the interplay between stress responses, ABA signaling, and post-transcriptional regulation in plants.</p

An exploration of the determinants for decision to migrate existing resources to cloud computing using an integrated TOE-DOI model

Migrating existing resources to cloud computing is a strategic organisational decision that can be difficult. It requires the consideration and evaluation of a wide range of technical and organisational aspects. Although a significant amount of attention has been paid by many industrialists and academics to aid migration decisions, the procedure remains difficult. This is mainly due to underestimation of the range of factors and characteristics affecting the decision for cloud migration. Further research is needed to investigate the level of effect these factors have on migration decisions and the overall complexity. This paper aims to explore the level of complexity of the decision to migrate the cloud. A research model based on the diffusion of innovation (DOI) theory and the technology-organization-environment (TOE) framework was developed. The model was tested using exploratory and confirmatory factor analysis. The quantitative analysis shows the level of impact of the identified variables on the decision to migrate. Seven determinants that contribute to the complexity of the decisions are identified. They need to be taken into account to ensure successful migration. This result has expanded the collective knowledge about the complexity of the issues that have to be considered when making decisions to migrate to the cloud. It contributes to the literature that addresses the complex and multidimensional nature of migrating to the cloud

Effects of hospital facilities on patient outcomes after cancer surgery: an international, prospective, observational study

Background Early death after cancer surgery is higher in low-income and middle-income countries (LMICs) compared with in high-income countries, yet the impact of facility characteristics on early postoperative outcomes is unknown. The aim of this study was to examine the association between hospital infrastructure, resource availability, and processes on early outcomes after cancer surgery worldwide.Methods A multimethods analysis was performed as part of the GlobalSurg 3 study-a multicentre, international, prospective cohort study of patients who had surgery for breast, colorectal, or gastric cancer. The primary outcomes were 30-day mortality and 30-day major complication rates. Potentially beneficial hospital facilities were identified by variable selection to select those associated with 30-day mortality. Adjusted outcomes were determined using generalised estimating equations to account for patient characteristics and country-income group, with population stratification by hospital.Findings Between April 1, 2018, and April 23, 2019, facility-level data were collected for 9685 patients across 238 hospitals in 66 countries (91 hospitals in 20 high-income countries; 57 hospitals in 19 upper-middle-income countries; and 90 hospitals in 27 low-income to lower-middle-income countries). The availability of five hospital facilities was inversely associated with mortality: ultrasound, CT scanner, critical care unit, opioid analgesia, and oncologist. After adjustment for case-mix and country income group, hospitals with three or fewer of these facilities (62 hospitals, 1294 patients) had higher mortality compared with those with four or five (adjusted odds ratio [OR] 3.85 [95% CI 2.58-5.75]; p&lt;0.0001), with excess mortality predominantly explained by a limited capacity to rescue following the development of major complications (63.0% vs 82.7%; OR 0.35 [0.23-0.53]; p&lt;0.0001). Across LMICs, improvements in hospital facilities would prevent one to three deaths for every 100 patients undergoing surgery for cancer.Interpretation Hospitals with higher levels of infrastructure and resources have better outcomes after cancer surgery, independent of country income. Without urgent strengthening of hospital infrastructure and resources, the reductions in cancer-associated mortality associated with improved access will not be realised

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Proximal contact tightness of class bulk-fill composite resin restorations: An in vitro study.

This study investigated the effect of bulk-fill composites on proximal contact tightness (PCT) of composite restorations using different matrix systems. 150/standardized-MO-ivorine cavity preparations were divided into 5 groups; Smart Dentin Replacement (SDR), SonicFill (SF), Tetric EvoCeram Bulk-Fill (TEB), G-aenial Universal Flo (GF) and Tetric EvoCeram (TE). Each group was subdivided into 3 sub-groups (n=10); Dixieland band in Tofflemire retainer, FenderMate and Palodent plus matrix systems. PCT was measured 24 h post-curing using Tooth Pressure Meter. PCT means were calculated and statistically-analyzed using ANOVA and Tukey's post-hoc test (p<0.05). Means and SD of PCT for Tofflemire subgroup were: 1.75(0.13), 3.21(0.1), 3.06(0.19), 2.49(0.21) and 3.18(0.1) for (SDR), (SF), (TEB), (GF) and (TE), respectively. Using FenderMate, values were: 1.87(0.08), 3.35(0.12), 3.17(0.16), 2.64(0.1) and 3.26(0.11) for (SDR), (SF), (TEB), (GF) and (TE), respectively, while with Palodentplus; 3.16(0.17), 4.23(0.11), 4.1(0.1), 3.46(0.17) and 3.98(0.1) for (SDR), (SF), (TEB), (GF) and (TE), respectively. ANOVA revealed significant differences (p<0.05) between all samples except between (SF), (TEB) and (TE) and also significant difference between Palodentplus and two subgroups. Effect of bulk-fill composites on PCT is material dependent. Separation ring is recommended for proper PCT

Effects of Ce-Dy rare earths co-doping on various features of Ni-Co spinel ferrite microspheres prepared via hydrothermal approach

The effects of Ce-Dy co-doping on the crystal structure, optical, dielectric, magnetic properties, and hyperfine interactions of Ni-Co spinel ferrite microspheres synthesized hydrothermally have been studied. A series of ferrites with the general formula Ni0.5-Co0.5CexDyxFe2-2xO4 were synthesized with x values ranging from 0.00 to 0.10. The phase, crystallinity, and morphology of ferrite microspheres were analyzed by X-ray powder diffractometry (XRD), scanning and transmission electron microscopes (SEM and TEM), respectively. The structural analyses of the synthesized ferrite microspheres confirmed their high purity and cubic crystalline phase. The Diffuse reflectance spectroscopic (DRS) measurements were presented to calculate direct optical energy band gaps (E-g) and is found in the range 1.63 eV - 1.84 eV. Fe-57 Mossbauer spectroscopy showed that the hyperfine magnetic field of tetrahedral (A) and octahedral (B) sites decreased with the substitution of Dy3+-Ce3+ ions that preferrentially occupy the B site. The impact of the rare-earth content (x) on the magnetic features of the prepared NiCo ferrite microspheres was investigated by analyzing M-H loops, which showed soft ferrimagnetism. The magnetic features illustrate a great impact of the incorporation of Ce3+-Dy3+ ions within the NiCo ferrite structure. The saturation magnetization (M-s), remanence (M-r), and coercivity (H-c) increased gradually with increasing Ce-Dy content. At x = 0.04, M-s, M-r, and H-c attain maximum values of about 31.2 emu/g, 11.5 emu/g, and 512.4 Oe, respectively. The Bohr magneton (n(B)) and magneto-crystalline anisotropy constant (K-eff) were also determined and evaluated with correlation to other magnetic parameters. Further increase in Ce3+-Dy3+ content (i.e., x >= 0.06) was found to decrease M-s, M-r, and H-c values. The variations in magnetic parameters (M-s, M-r, and H-c) were largely caused by the surface spins effect, the variations in crystallite/particle size, the distribution of magnetic ions into the different sublattices, the evolutions of magneto-crystalline anisotropy, and the variations in the magnetic moment (n(B)). The squareness ratios were found to be lower than the predicted theoretical value of 0.5 for various samples, indicating that the prepared Ce-Dy substituted NiCo ferrite microspheres are composed of NPs with single-magnetic domain (SMD). Temperature and frequency-dependent electrical and dielectric measurements have been done to estimate the ac/dc conductivity, dielectric constant, and tangent loss values for all the samples. The ac conductivity measurements confirmed the power-law rules, largely dependent on Ce-Dy content. Impedance analysis stated that the conduction mechanisms in all samples are mainly due to the grains-grain boundaries. The dielectric constant of NiCo ferrite microspheres give rise to normal dielectric distribution, with the frequency depending strongly on the Ce-Dy content. The observed variation in tangential loss with frequency can be attributed to the conduction mechanism in ferrites, like Koop's phenomenological model. (C) 2021 The Author(s). Published by Elsevier B.V.Deanship for Scientific Research of Imam Abdulrahman Bin Faisal University [2020-164-IRMC]The authors acknowledge the financial supports provided by the Deanship for Scientific Research of Imam Abdulrahman Bin Faisal University (Project application No. 2020-164-IRMC)