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At least 649 records · Page 36

High-efficiency Agrobacterium-mediated transformation of Norway spruce (Picea abies) and loblolly pine (Pinus taeda)

Agrobacterium-mediated gene transfer is the method of choice for many plant biotechnology laboratories; however, large-scale use of this organism in conifer transformation has been limited by difficult propagation of explant material, selection efficiencies and low transformation frequency. We have analyzed co-cultivation conditions and different disarmed strains of Agrobacterium to improve transformation. Additional copies of virulence genes were added to three common disarmed strains. These extra virulence genes included either a constitutively active virG or extra copies of virG and virB, both from pTiBo542. In experiments with Norway spruce, we increased transformation efficiencies 1000-fold from initial experiments where little or no transient expression was detected. Over 100 transformed lines expressing the marker gene beta-glucuronidase (GUS) were generated from rapidly dividing embryogenic suspension-cultured cells co-cultivated with Agrobacterium. GUS activity was used to monitor transient expression and to further test lines selected on kanamycin-containing medium. In loblolly pine, transient expression increased 10-fold utilizing modified Agrobacterium strains. Agrobacterium-mediated gene transfer is a useful technique for large-scale generation of transgenic Norway spruce and may prove useful for other conifer species.

NASA Discipline Plant Biology↗

Laser Hardening Prediction Tool Based On a Solid State Transformations Numerical Model

This paper presents a tool to predict hardening layer in selective laser hardening processes where laser beam heats the part locally while the bulk acts as a heat sink.The tool to predict accurately the temperature field in the workpiece is a numerical model that combines a three dimensional transient numerical solution for heating where is possible to introduce different laser sources. The thermal field was modeled using a kinetic model based on Johnson-Mehl-Avrami equation. Considering this equation, an experimental adjustment of transformation parameters was carried out to get the heating transformation diagrams (CHT). With the temperature field and CHT diagrams the model predicts the percentage of base material converted into austenite. These two parameters are used as first step to estimate the depth of hardened layer in the part.The model has been adjusted and validated with experimental data for DIN 1.2379, cold work tool steel typically used in mold and die making industry. This steel presents solid state diffusive transformations at relative low temperature. These transformations must be considered in order to get good accuracy of temperature field prediction during heating phase. For model validation, surface temperature measured by pyrometry, thermal field as well as the hardened layer obtained from metallographic study, were compared with the model data showing a good adjustment.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗

7.2 kV Three-Port SiC Single-Stage Current-Source Solid-State Transformer With 90 kV Lightning Protection

This article proposes a multiport modular single-stage current-source solid-state transformer (SST) for applications like photovoltaic, energy storage integration, electric vehicle fast charging, data center, etc. The 7.2 kV 50 kVA current-source SST consists of five input-series output-parallel modules, each based on 3.3 kV SiC reverse-blocking MOSFET-plus-diode modules. The proposed SST has some unique features. First, compared to the voltage-source or matrix converter-based SSTs, the current-source SST has a unique advantage of single-stage isolated AC/DC or AC/AC conversion with an inductive DC link, but no medium-voltage (MV) AC experiments have been reported. This article for the first time demonstrates MV AC current-source SST up to 7.5 kV peak. Second, the multiport SST has a buffer port for active power decoupling (APD) or energy storage integration. The double-line-frequency power ripple from single-phase AC grid normally results in a large capacitor size in MV SSTs. The APD scheme is proposed in MV applications for the first time to enable a reduced DC link and the electrolytic capacitor-less SST with high reliability. Third, as a direct grid-connected converter without line-frequency transformer, insulation and protection are critical. A medium-frequency transformer design passes 55 kV basic-insulation level (BIL) and 60 kV high potential dielectrics withstand test with only 0.09% leakage inductance. Importantly, a lightning protection scheme is presented to protect the SST itself from 90 kV BIL impulse. Fourth, the proposed current-source SST topology is a modular soft-switching solid-state transformer (M-S4T) with full-range zero-voltage switching and controlled dv/dt for low electromagnetic interference. Furthermore, these concepts are verified in a three-port M-S4T prototype with forced oil cooling under single-module, stacked-module, steady-state, and dynamic operations.

14 SOLAR ENERGY↗

Visualizing Crystallization Dynamics and Transformation Pathways of Disordered Rocksalt Oxides During Thermally Activated Sol–Gel Synthesis

Sol–gel synthesis is a wet-chemical processing route for fabricating functional materials with control over composition and microstructure at relatively low temperatures compared to conventional solid-state synthesis. While sol–gel process initiates with intermixed molecular precursors, the early-stage nucleation pathways are insufficiently understood. Here, in this study, the chemical and structural transformation of ion disordered rocksalt (DRX) Li 1.2 Mn 0.4 Ti 0.4 O 2 (LMTO), a promising cathode material for lithium batteries, is studied by multiscale characterizations. In situ heating transmission electron microscopy (TEM) using a liquid cell visualizes and identifies crystallization pathways at the nanoscale. While some regions follow a classical multi-step transition through thermodynamically stable intermediates, others exhibit a kinetic shortcut via a localized amorphous matrix to directly form the DRX structure. Macroscale Fourier transform infrared spectroscopy corroborates the findings and reveals that transition metal ions are more strongly incorporated into the acetate-coordinated network than lithium. Although in situ heating TEM captures diverse local transformation pathways, in situ synchrotron X-ray diffraction indicates that the macroscopic transformation proceeds predominantly through spinel LMTO and lithium titanates toward DRX-LMTO. The findings uncover the spatiotemporal chemical and structural transformations in sol–gel derived DRX-LMTO materials, and call for fine-tuning of such sol–gel chemistries to manipulate the crystallization pathways and achieve target material homogeneity more efficiently.

cathode material↗

Agrobacterium ‐Mediated Transformation of Brachypodium distachyon

Abstract Brachypodium distachyon is an excellent model system for the grasses and has been adopted as a research organism by many laboratories around the world. It has all of the biological traits required for a model system, including small stature, short life cycle, small genome, simple growth requirements, and a close relationship to major crop plants (cereals). In addition, numerous resources have been developed for working with this species, including genome sequences for many lines, sequenced mutant collections, and a large, freely available germplasm collection. Fortunately, among grasses B. distachyon is one of the most easily transformed species, an absolute necessity for a model system. Agrobacterium ‐mediated transformation is the preferred method to transform plants because it usually results in simple insertions of target DNA. In this article, we describe a method for Agrobacterium ‐mediated transformation of the inbred B. distachyon lines Bd21 and Bd21‐3. Embryogenic callus induced from immature embryos is co‐cultivated with Agrobacterium tumefaciens strain AGL1 or Agrobacterium rhizogenes strain 18r12v. Hygromycin and paromomycin are used as selective agents, with comparable transformation efficiencies (defined as the percentage of co‐cultivated callus that produce transgenic plants) of 40% to 70%. It takes 20 to 30 weeks to obtain T 1 seeds starting from the initial step of dissecting out immature embryos. This protocol has been shown to be efficient and facile in several studies that resulted in the creation of over 22,000 T‐DNA mutants. © 2019 by John Wiley & Sons, Inc.

Chen, Fengjuan↗

Stability of linear systems in second-order form based on structure preserving similarity transformations

This paper deals with two stability aspects of linear systems of the form I ¨ x +B˙ x +Cx = 0 given by the triple (I;B;C). A general transformation scheme is given for a structure and Jordan form preserving transformation of the triple. We investigate how a system can be transformed by suitable choices of the transformation parameters into a new system (I;B1;C1) with a symmetrizable matrix C1. This procedure facilitates stability investigations. We also consider systems with a Hamiltonian spectrum which discloses marginal stability after a Jordan form preserving transformation.

Stoustrup, Jakob↗

Agrobacterium tumefaciens-mediated transformation of oleaginous yeast Lipomyces species

Background: Because of interest in the production of renewable bio-hydrocarbon fuels, various living organisms have been explored for their potential use in producing fuels and chemicals. The oil-producing (oleaginous) yeast Lipomyces starkeyi is the subject of active research regarding the production of lipids using a wide variety of carbon and nutrient sources. The genome of L. starkeyi has been published, which opens the door to production strain improvements using the tools of synthetic biology and metabolic engineering. However, using these tools for strain improvement requires the establishment of effective and reliable transformation methods with suitable selectable markers (antibiotic resistance or auxotrophic marker genes) and the necessary genetic elements (promoters and terminators) for expression of introduced genes. Chemical-based methods have been published, but suffer from low efficiency or the requirement for targeting to rRNA loci. To address these problems, Agrobacterium-mediated transformation was investigated as an alternative method for L. starkeyi and other Lipomyces species. Results: In this study, Agrobacterium-mediated transformation was demonstrated to be effective in the transformation of both L. starkeyi and other Lipomyces species and that the introduced DNA can be reliably integrated into the chromosomes of these species. The gene deletion of Ku70 and Pex10 was also demonstrated in L. starkeyi. In addition to the bacterial antibiotic selection marker gene hygromycin B phosphotransferase, the bacterial -glucuronidase reporter gene under the control of L. starkeyi translation elongation factor 1 promoter was also stably expressed in seven different Lipomyces species. Conclusion: The results from this study clearly demonstrate that Agrobacterium-mediated transformation is a reliable genetic tool for gene deletion and integration and expression of heterologous genes in L. starkeyi and other Lipomyces species.

Lipomyces doorenjongii↗

Improved high-throughput screening technique to rapidly isolate Chlamydomonas transformants expressing recombinant proteins

Abstract The single-celled eukaryotic green alga Chlamydomonas reinhardtii has long been a model system for developing genetic tools for algae, and is also considered a potential platform for the production of high-value recombinant proteins. Identifying transformants with high levels of recombinant protein expression has been a challenge in this organism, as random integration of transgenes into the nuclear genome leads to low frequency of cell lines with high gene expression. Here, we describe the design of an optimized vector for the expression of recombinant proteins in Chlamydomonas , that when transformed and screened using a dual antibiotic selection, followed by screening using fluorescence activated cell sorting (FACS), permits rapid identification and isolation of microalgal transformants with high expression of a recombinant protein. This process greatly reduces the time required for the screening process, and can produce large populations of recombinant algae transformants with between 60 and 100% of cells producing the recombinant protein of interest, in as little as 3 weeks, that can then be used for whole population sequencing or individual clone analysis. Utilizing this new vector and high-throughput screening (HTS) process resulted in an order of magnitude improvement over existing methods, which normally produced under 1% of algae transformants expressing the protein of interest. This process can be applied to other algal strains and recombinant proteins to enhance screening efficiency, thereby speeding up the discovery and development of algal-derived recombinant protein products. Key points • A protein expression vector using double-antibiotic resistance genes was designed • Double antibiotic selection causes fewer colonies with more positive for phenotype • Coupling the new vector with FACS improves microalgal screening efficiency > 60%

59 BASIC BIOLOGICAL SCIENCES↗

An asymmetric hybrid cryptosystem using equal modulus and random decomposition in hybrid transform domain

In this paper, an asymmetric hybrid cryptosystem with coherent superposition, equal modulus and random decomposition in hybrid transform domain is proposed. To further strengthen the security of the cryptosystem, a hyperchaotic system is used as a pixel-swapping procedure. The hybrid transform is generated by utilizing fractional Fourier transform of various orders and Walsh transform. The hyperchaotic framework’s parameters and starting conditions alongside the fractional orders of the fractional Fourier transform extend the key-space and consequently give extra strength to the proposed cryptosystem. The designed cryptosystem has an extended key-space to avoid any brute-force attack and is non-linear in nature. The scheme is validated on gray-scale images. Computer based simulations have been performed to verify the validity and the performance of the proposed cryptosystem against different types of attacks. Results demonstrate that the proposed cryptosystem not only offers higher protection against noise attacks but is also invulnerable to special attack.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Temperature-dependent micromechanical behavior of medium-Mn transformation-induced-plasticity steel studied by in situ synchrotron X-ray diffraction

The temperature-dependent micromechanical behavior of medium-Mn transformation-induced-plasticity (TRIP) steel with a nominal chemical composition of Fe-0.1C-10Mn-2Al (mass%) fabricated by intercritical annealing 650 ºC for 1 h after cold-rolling, was investigated using in situ high-energy X-ray diffraction (HE-XRD) with uniaxial tensile tests at temperatures of 100, 25 and -50 ºC. We find that as the deformation temperature decreases, the Luders strain decreases and more austenite transforms to martensite during Luders band propagation. The Luders bands are associated with sudden changes of lattice strain in austenite. At 100 ºC, austenite is too stable to transform to martensite, resulting in limited work-hardening capability and a relatively low strain to failure. At 25 ºC, the austenite is found to transform in bursts during applied loading. These transformations correlate with stepwise peak broadening in the austenite phase and are attributed to Portevin-Le Chatelier (PLC) band propagation. At -50 ºC, we then observe a more intense TRIP effect which suppresses PLC band formation and leads to a high ultimate tensile strength. In addition to the TRIP effect, we find that work hardening in the deformation-induced martensite phase plays an important role in the plastic stability at low temperature, by accommodating stress compatibility between grains and phases. Our studies provide the first direct experimental evidence for the existence of large stress concentrations in the austenite phase near the Luders band propagation front. These observations help to deepen understanding of the complex temperature-dependent micromechanical behaviors of advanced medium-Mn TRIP steels.

36 MATERIALS SCIENCE↗

C6 glioma-conditioned medium induces malignant transformation of mesenchymal stem cells: Possible role of S100B/RAGE pathway

Highlights: • C6 glioma-conditioned medium induces MSCs malignant transformation. • The transformed cells are tumorigenesis and phenotypically distinct from MSCs. • GCM increases MSCs RAGE, Akt1 and STAT3 expression. • S100B increases MSCs RAGE, Akt1, STAT3 and proliferation, migration and invasion. • RAGE inhibitor abolish S100B induced Akt1, STAT3 activation and cellular functions. Mesenchymal stem cells (MSCs) have been widely studied as an attractive therapeutic agent for the treatment of tumors. However, the adverse effects of the tumor paracrine factors who affect MSCs are still unclear. In this study, we report for the first time that C6 glioma-conditioned medium (GCM) induces malignant transformation of MSCs. In contrast to MSCs, the transformed mesenchymal stem cells (TMCs) exhibited tumor cell characterizations in vitro and highly tumorigenic in vivo. Furthermore, GCM and recombinant S100B increased receptor for advanced glycation end products (RAGE) and its downstream Akt1, STAT3 genes expression as well as phosphorylation and transcriptional activation. Finally, blockage of S100B-RAGE interaction by RAGE inhibitor FPS-ZM1 attenuated GCM and S100B-induced Akt1, STAT3 activation, abolished its cell proliferation, migration and invasion actions. Together, these results suggest that the RAGE pathway may play a possible role in malignant transformation procedure of MSCs, and that this process may be mediated through S100B.

60 APPLIED LIFE SCIENCES↗

Stable plastid transformation of rice, a monocot cereal crop

Highlights: • The homoplastomic rice plant was firstly obtained by plastid transformation. • The results was confirmed by both molecular evidences and detecting localization of GFP. • The results will be helpful to transform plastid genome of monocotyledonous crops with recalcitrant nature. Rice is one of the most important cereal crops and its biotechnology has been pursued to meet the food demand of ever-growing global population. Rice plastid transformation has been a great challenge to achieve homoplastomic plants due to its low efficiency of regeneration. In this experiment, Japonica rice line 19 was chosen to be the receptor for plastid transformation. A vector harboring smGFP gene was constructed and transferred into rice plastid genome by bombardment. The resistant callus was obtained after long-lasting multiple selections and proved to be in homoplastomic status by molecular testing. The plantlet was regenerated from homoplastomic callus and grown to seeding stage. This is the first case so far to achieve the homoplastomic rice and will be helpful to transform plastid genome of monocotyledonous crops with recalcitrant nature.

60 APPLIED LIFE SCIENCES↗

Understanding the long-term influence of EIA on organisational learning and transformation

This research is an attempt to verify the notion postulated by Robert Bartlett and Lynton Caldwell that the full benefits of environmental impact assessment (EIA) would take decades to be realized. While EIA is intended to directly influence decision-making regarding new development proposals, the process is also expected to lead to organisational learning and transformation over time. Our aim was to examine the influence of EIA on a single Western Australian proponent with sustained experience in the process to understand how EIA is used within the organisation and to seek evidence of transformation of the organisation's purpose and mission. The research reviewed literature in order to identify key influences of EIA on organisations, along with semi-structured staff interviews and document analysis for the case study organisation. Ascertaining causality that involvement in EIA processes influences or effects organisational learning and transformation is a challenge in the face of other societal events. Document analysis and interviewee data indicates that the action-forcing nature of EIA did influence proponent behavior through the creation of internal processes seeking to ensure robust design of new projects that would satisfy environmental protection expectations, without the need to trigger formal EIA. Evidence of EIA values and thinking were apparent within internal documentation, including the evolving mission statement. Our research indicates that participation in the EIA process can positively influence organisational learning and transformation by guiding internal change for decision-making. - Highlights: • The long-term influence of EIA on a proponent organisation is investigated. • EIA promotes internal organisational learning and transformation. • Analysis of mission statements can indicate the influence of EIA on organisations. • Organisations aligned with EIA values can reduce the need to engage in formal EIA.

54 ENVIRONMENTAL SCIENCES↗

Mercury transformation processes in nature: Critical knowledge gaps and perspectives for moving forward

We report the transformation of mercury (Hg) in the environment plays a vital role in the cycling of Hg and its risk to the ecosystem and human health. Of particular importance are Hg oxidation/reduction and methylation/demethylation processes driven or mediated by the dynamics of light, microorganisms, and organic carbon, among others. Advances in understanding those Hg transformation processes determine our capacity of projecting and mitigating Hg risk. Here, we provide a critical analysis of major knowledge gaps in our understanding of Hg transformation in nature, with perspectives on approaches moving forward. Our analysis focuses on Hg transformation processes in the environment, as well as emerging methodology in exploring these processes. Future avenues for improving the understanding of Hg transformation processes to protect ecosystem and human health are also explored.

54 ENVIRONMENTAL SCIENCES↗

Bundling measures for food systems transformation: a global, multimodel assessment

Background Current food systems leave one in ten individuals at risk of hunger while driving unsustainable environmental impacts. Inaction risks further exacerbating negative impacts on both human and planetary health. These challenges emerge from complex system interactions, requiring approaches that engage with this complexity and consider how transformation measures interact across food systems. We aimed to quantify the magnitude and uncertainty of the impacts of key food systems transformation measures both individually and in a bundle using an ensemble of global economic models. Methods In this global multimodel assessment, we applied an ensemble of ten state-of-the-art global economic models to evaluate the potential of four key measures in transforming food systems: increasing agricultural productivity, halving food loss and waste, shifting towards healthier diets, and economy-wide climate mitigation policies aligned with limiting warming to 1·5°C. The scenarios used a middle-of-the-road shared socioeconomic pathway for population and gross domestic product growth, climate impact data from Jägermeyr and colleagues, Thornton and colleagues, and Nelson and colleagues, and dietary targets based on the EAT–Lancet healthy reference diet, with model simulations conducted from 2020 to 2050. We then assessed the effect of these measures in isolation and in combination in a bundled scenario. To further understand the interactions between these measures, we conducted a decomposition analysis that distinguishes between the individual effects of a measure (effect when implemented alone), total effects (its contribution within the bundle), and interaction effects (the difference between total and individual effects). This approach aimed to show complementarities and trade-offs that emerge when multiple measures are implemented simultaneously. Findings Our analysis showed that individual measures in isolation are insufficient to achieve high-level environmental objectives and might generate unintended consequences. In contrast, bundling measures produces co-benefits: avoiding 50% of projected agricultural greenhouse gas emissions by 2050 and almost 20% of anticipated land conversion, while moderating food price increases associated with ambitious climate change mitigation policies. Our decomposition analysis further shows that measures can have varying effects across different dimensions. Although dietary shifts and climate mitigation policies are the largest drivers of environmental benefits (each contributing to a median decline of >10 percentage points in non-CO 2 emissions and 5 percentage points in agricultural land use globally), productivity improvements and reducing food loss and waste play essential roles in moderating price increases (each contributing to a median decline of >5 percentage points in average prices). Interpretation This study highlights the importance of implementing coordinated approaches to food system transformation and climate change mitigation rather than relying on isolated interventions. Comprehensive transformation requires understanding how supply-side and demand-side changes can interact with climate mitigation policies, enabling policy makers to design intervention packages that maximise benefits while minimising trade-offs across environmental, economic, and social dimensions.

Sundiang, Marina [Cornell Univ., Ithaca, NY (Unite↗

The effect of annealing on the transformation and the microstructure of Mn{sub 1−x}Cr{sub x}CoGe alloys

In this work the effect of different thermal treatments on the transformation behavior of Mn{sub 1−x}Cr{sub x}CoGe alloys, with x = 0.15 and 0.20 has been analyzed. The changes in the transformation temperatures have been studied by differential scanning calorimetry (DSC). The results show that the structural transition temperature depends on the previous annealing. However, under the same heat treatment no significant change is observed on the transformation temperatures when replacing Mn by Cr. The microstructural evolution has been monitored using in-situ X-ray diffraction and transmission electron microscopy. The effect of an applied magnetic field on the transformation has been explored by SQUID magnetometry. Two different magnetic transitions are found: a first order paramagnetic (PM) to ferromagnetic (FM) corresponding to the transformation observed by calorimetry and a re-entrant spin glass to ferromagnetic transition. - Highlights: • The annealing conditions determine the magnetostructural transition. • A laminar microstructure along preferential planes observed by TEM • A RSG to ferromagnetic transition at low temperatures is detected.

36 MATERIALS SCIENCE↗

Positive and negative effects of dielectric breakdown in transformer oil based magnetic fluids

The transformer oil based magnetic fluids can be considered as the next-generation insulation fluids because they offer exciting new possibilities to enhance dielectric breakdown voltage as well as heat transfer performance compared to pure transformer oils. In this study, we have investigated the dielectric breakdown strength of the fluids with the various volume concentrations of nanoparticles in accordance with IEC 156 standard and have tried to find the reason for changing the dielectric breakdown voltage of the fluids from the magnetic field analysis. It was found that the dielectric breakdown voltage of pure transformer oil is around 12 kV with the gap distance of 1.5 mm. In the case of our transformer oil-based magnetic fluids with 0.08% < Φ < 0.6% (Φ means the volume concentration of magnetic nanoparticles), the dielectric breakdown voltage shows above 40 kV, which is 3.3 times higher positively than that of pure transformer oil. Negatively in the case when the volume concentration of magnetic nanoparticles is above 0.65%, the dielectric breakdown voltage decreases reversely. From the magnetic field analysis, the reason might be considered as two situations: the positive is for the conductive nanoparticles dispersed well near the electrodes, which play an important role in converting fast electrons to slow negatively charged particles, and the negative is for the agglomeration of the particles near the electrodes, which leads to the breakdown initiation.

36 MATERIALS SCIENCE↗

Arginine inhibits the malignant transformation induced by interferon-gamma through the NF-κB-GCN2/eIF2α signaling pathway in mammary epithelial cells in vitro and in vivo

Highlights: • Arginine could antagonize the malignant transformation of mammary epithelial cells induced by IFN-γ both in vitro and in vivo. • We found fewer arginine and more IFN-γ in patients with breast cancer than those with benign breast disease, • The expressions of arginine and IFN-γ are related closely to TNM stage, ER (-/+), PR (-/+) and so on. Breast cancer is the most common female malignant tumors in the world. It seriously affects women's physical and mental health and the leading cause of cancer death among women. Our previous study demonstrated that diet-derived IFN-γ promoted the malignant transformation of primary bovine mammary epithelial cells by accelerating arginine depletion. The current study aimed to explore whether arginine addition could inhibit the degree of malignant transformation and its molecular mechanism. The results indicate that arginine addition could alleviate the malignant transformation of mammary epithelial cells induced by IFN-γ, including reducing cell proliferation, cell migration and colony formation, through the NF-κB-GCN2/eIF2α pathway. The in vivo experiments also consistently confirmed that arginine supplementation could significantly inhibit tumor growth in tumor-bearing mice. Furthermore, the investigation of the clinical data also revealed that the plasma or tissue from human breast cancer patients owned lower arginine level and higher IFN-γ level than that from patients with benign breast disease, showing IFN-γ may be a potential control target. Our findings demonstrate that arginine supplement could antagonize the malignant transformation of mammary epithelial cells induced by IFN-γ (nutritionally induced) both in vitro and in vivo, and IFN-γ was higher in breast cancer women. This might provide a novel strategy for the prevention and treatment of breast cancer regarding to nutrition.

60 APPLIED LIFE SCIENCES↗