Preparation of monoclonal antibody against rhoifolin and its application in enzyme-linked immunosorbent assay of rhoifolin and diosmin.

Both rhoifolin and diosmin belong to flavonoids, which are widely present in citrus. Diosmin is not only used in the medical field in the world, but also used as a dietary supplement in the United States. Rhoifolin has a similar structure to diosmin and also exhibits antioxidant and anti-inflammatory properties. In this study, an anti-rhoifolin monoclonal antibody was prepared and an indirect competitive enzyme-linked immunosorbent assay (icELISA) method was established. The half-maximal inhibitory concentration (IC50) of icELISA was determined to be 4.83 ng/mL, and the detection range was 0.97-33.87 ng/mL. The results of UPLC-MS/MS and icELISA generally demonstrate consistency. Moreover, by exploiting the cross-reactivity of the antibody, diosmin in tablets can be detected by icELISA. The results demonstrate that the developed method has good accuracy, reproducibility, and broad application prospects.

LEAFY and WAPO1 jointly regulate spikelet number per spike and floret development in wheat.

In wheat, the transition of the inflorescence meristem to a terminal spikelet (IM→TS) determines the spikelet number per spike (SNS), an important yield component. In this study, we demonstrate that the plant-specific transcription factor LEAFY (LFY) physically and genetically interacts with WHEAT ORTHOLOG OF APO1 (WAPO1) to regulate SNS and floret development. Loss-of-function mutations in either or both genes result in significant and similar reductions in SNS, as a result of a reduction in the rate of spikelet meristem formation per day. SNS is also modulated by significant genetic interactions between LFY and the SQUAMOSA MADS-box genes VRN1 and FUL2, which promote the IM→TS transition. Single-molecule fluorescence in situ hybridization revealed a downregulation of LFY and upregulation of the SQUAMOSA MADS-box genes in the distal part of the developing spike during the IM→TS transition, supporting their opposite roles in the regulation of SNS in wheat. Concurrently, the overlap of LFY and WAPO1 transcription domains in the developing spikelets contributes to normal floret development. Understanding the genetic network regulating SNS is a necessary first step to engineer this important agronomic trait.

Mitochondria-Targeted Natural Antioxidant Nanosystem for Diabetic Vascular Calcification Therapy.

The development of nanotherapy targeting mitochondria to alleviate oxidative stress is a critical therapeutic strategy for vascular calcification (VC) in diabetes. In this study, we engineered mitochondria-targeted nanodrugs (T4O@TPP/PEG-PLGA) utilizing terpinen-4-ol (T4O) as a natural antioxidant and mitochondrial protector, PEG-PLGA as the nanocarrier, and triphenylphosphine (TPP) as the mitochondrial targeting ligand. In vitro assessments demonstrated enhanced cellular uptake of T4O@TPP/PEG-PLGA, with effective mitochondrial targeting. This nanodrug successfully reduced oxidative stress induced by high glucose levels in vascular smooth muscle cells. In vivo studies showed prolonged retention of the nanomaterials in the thoracic aorta for up to 24 h. Importantly, experiments in diabetic VC models underscored the potent antioxidant properties of T4O@TPP/PEG-PLGA, as evidenced by its ability to mitigate VC and restore mitochondrial morphology. These results suggest that these nanodrugs could be a promising strategy for managing diabetic VC.

Data imbalance in cardiac health diagnostics using CECG-GAN.

Heart disease is the world's leading cause of death. Diagnostic models based on electrocardiograms (ECGs) are often limited by the scarcity of high-quality data and issues of data imbalance. To address these challenges, we propose a conditional generative adversarial network (CECG-GAN). This strategy enables the generation of samples that closely approximate the distribution of ECG data. Additionally, CECG-GAN addresses waveform jitter, slow processing speeds, and dataset imbalance issues through the integration of a transformer architecture. We evaluated this approach using two datasets: MIT-BIH and CSPC2020. The experimental results demonstrate that CECG-GAN achieves outstanding performance metrics. Notably, the percentage root mean square difference (PRD) reached 55.048, indicating a high degree of similarity between generated and actual ECG waveforms. Additionally, the Fréchet distance (FD) was approximately 1.139, the root mean square error (RMSE) registered at 0.232, and the mean absolute error (MAE) was recorded at 0.166.

In Situ Synthesis of an Immune-Checkpoint Blocker from Engineered Bacteria Elicits a Potent Antitumor Response.

Immune-checkpoint blockade (ICB) reinvigorates T cells from exhaustion and potentiates T-cell responses to tumors. However, most patients do not respond to ICB therapy, and only a limited response can be achieved in a "cold" tumor with few infiltrated lymphocytes. Synthetic biology can be used to engineer bacteria as controllable bioreactors to synthesize biotherapeutics in situ. We engineered attenuated Salmonella VNP20009 with synthetic gene circuits to produce PD-1 and Tim-3 scFv to block immunosuppressive receptors on exhausted T cells to reinvigorate their antitumor response. Secreted PD-1 and Tim-3 scFv bound PD-1+ Tim-3+ T cells through their targeting receptors in vitro and potentiated the T-cell secretion of IFN-γ. Engineered bacteria colonized the hypoxic core of the tumor and synthesized PD-1 and Tim-3 scFv in situ, reviving CD4+ T cells and CD8+ T cells to execute an antitumor response. The bacteria also triggered a strong innate immune response, which stimulated the expansion of IFN-γ+ CD4+ T cells within the tumors to induce direct and indirect antitumor immunity.

Titanium Dioxide and Calcium Sulfate Whiskers Are Used for the Preparation of High Performance Polypropylene and Reduce White Pollution.

The anti-aging agent TiO2-polyacrylonitrile (PAN) and the mechanical strengthening agent CSW-PAN were prepared by radical polymerization using rutile nano-titanium dioxide (TiO2) and anhydrous calcium sulfate whisker (CSW) as raw materials. The structures of TiO2-PAN and CSW-PAN were characterized using Fourier transform infrared spectroscopy (FTIR) and X-ray photoelectron spectroscopy (XPS). Simultaneously, the mechanical properties, aging properties, and thermal stability of TiO2-PAN/CSW-PAN/polypropylene (PP) composites were studied, and the results showed that the surfaces of nano-titanium dioxide and calcium sulfate whiskers were successfully grafted with acrylonitrile. Owing to the introduction of new elements, such as acrylonitrile, nano-titanium dioxide and calcium sulfate whiskers have anti-aging properties. In comparison of the impact strength and tensile strength of TiO2-PAN/PP and TiO2-PAN/CSW-PAN/PP before aging, it can be proven that adding CSW-PAN can significantly enhance the mechanical properties of TiO2-PAN/CSW-PAN/PP. After 1000 h of aging, the tensile strength of the ternary composite TiO2-PAN/CSW-PAN/PP was 19.88 MPa when the addition amount of TiO2-PAN and CSW-PAN was 3%. Moreover, the impact strength of the ternary composite material TiO2-PAN/CSW-PAN/PP after 1000 h of aging is even better than that of non-aging pure PP materials, proving that the service life of improved PP products is extended, unnecessary waste and environmental pollution can be relieved, and the needs of specific engineering fields can be met.

Bioengineered Artificial Extracellular Vesicles Presenting PD-L1 and Gal-9 Ameliorate New-Onset Type 1 Diabetes.

An important factor in the development of type 1 diabetes (T1D) is the deficiency of inhibitory immune checkpoint ligands, specifically programmed cell death ligand 1 (PD-L1) and galectin-9 (Gal-9), in ß-cells. Therefore, modulation of pancreas-infiltrated T lymphocytes by exogenous PD-L1 or Gal-9 is an ideal approach for treating new-onset T1D. We genetically engineered macrophage cells to generate artificial extracellular vesicles (aEVs) overexpressing PD-L1 and Gal-9, which could restrict islet autoreactive T lymphocytes and protect ß-cells from destruction. Intriguingly, overexpression of Gal-9 stimulated macrophage polarization to the M2 phenotype with immunosuppressive attributes. Alternatively, both PD-L1- and Gal-9-presenting aEVs (PD-L1-Gal-9 aEVs) favorably adhered to T cells via the interaction of programmed cell death protein 1/PD-L1 or T-cell immunoglobulin mucin 3/Gal-9. Moreover, PD-L1-Gal-9 aEVs prominently promoted effector T-cell apoptosis and splenic regulatory T (Treg) cell formation in vitro. Notably, PD-L1-Gal-9 aEVs efficaciously reversed new-onset hyperglycemia in NOD mice, prevented T1D progression, and decreased the proportion and activation of CD4+ and CD8+ T cells infiltrating the pancreas, which together contributed to the preservation of residual ß-cell survival and mitigation of hyperglycemia.

Genomic insights into the origin and evolution of spelt (Triticum spelta L.) as a valuable gene pool for modern wheat breeding.

Spelt (Triticum aestivum ssp. spelta) is an important wheat subspecies mainly cultivated in Europe before the 20th century that has contributed to modern wheat breeding as a valuable genetic resource. However, relatively little is known about the origins and maintenance of spelt populations. Here, using resequencing data from 416 worldwide wheat accessions, including representative spelt wheat, we demonstrate that European spelt emerged when primitive hexaploid wheat spread to the west and hybridized with pre-settled domesticated emmer, the putative maternal donor. Genomic introgression regions from domesticated emmer confer spelt's primitive morphological characters used for species taxonomy, such as tenacious glumes and later flowering. We propose a haplotype-based "spelt index" to identify spelt-type wheat varieties and to quantify utilization of the spelt gene pool in modern wheat cultivars. This study reveals the genetic basis for the establishment of the spelt wheat subspecies in a specific ecological niche and the vital role of the spelt gene pool as a unique germplasm resource in modern wheat breeding.

ROS-responsive curcumin-encapsulated nanoparticles for AKI therapy via promoting lipid degradation in renal tubules.

Lipid accumulation is a factor contributing to the pathogenesis of acute kidney injury (AKI), yet there are currently no approved pharmacotherapies aside from adjuvant therapy. A developed reactive oxygen species (ROS)-responsive drug delivery system (NPSBG@Cur) was developed to deliver the autophagy activator curcumin (Cur) in order to alleviate AKI by activating autophagy and promoting lipid droplet degradation. The nanoparticles were shown to be ROS-responsive in the H2O2 medium and demonstrate ROS-responsive uptake in palmitate (PA)-induced oxidative stress-damaged cells. NPSBG@Cur was found to effectively inhibit lipid accumulation by autophagosome transport in kidney tubular cells. Additionally, in a mouse AKI model, NPSBG@Cur was observed to significantly ameliorate renal damage by activating autophagy flux and improving lipid transport. These results suggest that the ROS-responsive drug delivery system augmented the therapeutic effect of Cur on AKI by improving lipid metabolism through autophagy activation. Therefore, targeting lipid metabolism with NPSBG@Cur may be a promising AKI treatment strategy.

Preparation of high-performance anti-aging polypropylene by modified nano-TiO2 and calcium sulfate whisker grafted acrylonitrile composite PP.

By employing the radical polymerization method, acrylonitrile (AN) was grafted on the surface of nano titanium dioxide (TiO2), and the calcium sulfate whisker (CSW) was modified using the coupling agent KH570 to provide ultraviolet (UV)-absorption capacity. The prepared TiO2-PAN and CSW-PAN materials can improve the anti-aging performance and mechanical properties of polypropylene (PP) and meet the application requirements of high-performance polypropylene. Further, the obtained PP composites show prolonged service life and application scope, which can effectively reduce white waste and avoid both resource waste and environmental pollution.

Research on data imbalance in intrusion detection using CGAN.

To address the problems of attack category omission and poor generalization ability of traditional Intrusion Detection System (IDS) when processing unbalanced input data, an intrusion detection strategy based on conditional Generative Adversarial Networks (cGAN) is proposed. The cGAN generates attack samples that approximately obey the distribution pattern of input data and are randomly distributed within a certain bounded interval, which can avoid the redundancy caused by mechanical data widening. The experimental results show that the strategy has better performance indexes and stronger generalization ability in overall performance, which can solve insufficient classification performance and detection omission caused by unbalanced distribution of data categories and quantities.

A Sensor-Fault-Estimation Method for Lithium-Ion Batteries in Electric Vehicles.

In recent years, electric vehicles powered by lithium-ion batteries have developed rapidly, and the safety and reliability of lithium-ion batteries have been a paramount issue. Battery management systems are highly dependent on sensor measurements to ensure the proper functioning of lithium-ion batteries. Therefore, it is imperative to develop a suitable fault diagnosis scheme for battery sensors, to realize a diagnosis at an early stage. The main objective of this paper is to establish validated electrical and thermal models for batteries, and address a model-based fault diagnosis scheme for battery sensors. Descriptor proportional and derivate observer systems are applied for sensor diagnosis, based on electrical and thermal models of lithium-ion batteries, which can realize the real-time estimation of voltage sensor fault, current sensor fault, and temperature sensor fault. To verify the estimation effect of the proposed scheme, various types of faults are utilized for simulation experiments. Battery experimental data are used for battery modeling and observer-based fault diagnosis in battery sensors.

Dual-responsive antibiotic and baicalein co-delivery nanoparticles with enhanced synergistic antibacterial activity.

Globally, due to the rapid development of bacterial resistance, bacterial infections lead to significant mortality and morbidity which require efficient strategies to eradicate these infections. Herein, we prepared a dual-responsive synergistic drug delivery nanoparticle carrier (NPS@Bai/Cip), which responds to sub-acid bacterial microenvironments and targets phosphatase or phospholipase at infection sites. Nanoparticles surfaces were positively (10.0 mV) charged under acidic conditions, leading to good bacterial adhesion and enhanced drug accumulation. NPS@Bai/Cip showed good antibacterial and anti-biofilm activity against drug-resistant Pseudomonas aeruginosa. NPS@Bai/Cip could inhibit the biofilm formation via affecting the swimming, swarming, and twitching motilities of P. aeruginosa. NPS@Bai/Cip was used to treat drug-resistance P. aeruginosa-induced infection in rats by improving wound healing and reducing inflammatory responses. Thus, NPS@Bai/Cip functioned as an antibacterial and antibiofilm agent with good potential for treating bacteria-induced infections.

Synthesis and Characterization of a Novel DOPO-Based Flame Retardant Intermediate and Its Flame Retardancy as a Polystyrene Intrinsic Flame Retardant.

The research on intrinsic flame retardant has become a hot topic in the field of flame retardant. The synthesis of reactive flame-retardant monomer is one of the effective methods to obtain an intrinsic flame retardant. In addition, in view of the small molecular flame retardant easily migrates from the polymer during the use process, which leads to the gradual reduction of the flame retardant effect and even the gradual loss of flame retardant performance, and the advantages of atom transfer radical polymerization (ATRP) technology in polymer structure design and function customization, we first synthesized reactive flame retardant monomer 6-(hydroxymethyl)dibenzo[c,e][1,2]oxaphosphinine 6-oxide (FAA-DOPO), then synthesized polystyrene bromine (PS148-Br) macromolecular initiator by ATRP technology, and finally obtained block copolymer polystyrene-b-poly{6-(hydroxymethyl)dibenzo[c,e][1,2]oxaphosphinine 6-oxide} (PS-b-PFAA-DOPO) by the polymerization of FAA-DOPO initiated by macromolecular initiator PS148-Br by ATRP technology. The chemical structure of FAA-DOPO was characterized by 1D and 2D NMR (1H, 13C, DEPT 135, HSQC, COSY, NOE, and HMBC) spectra, Fourier transform infrared spectroscopy (FTIR), liquid chromatography-tandem mass spectrometry (LC-MS) and X-ray photoelectron spectroscopy (XPS). The chemical structure and molecular weight of PS-b-PFAA-DOPO were characterized by FTIR and gel permeation chromatography (GPC). The thermal and flame-retardant properties of PS-b-PFAA-DOPO were characterized by thermogravimetry analysis (TG), UL-94, limiting oxygen index (LOI), and microscale combustion calorimetry (MCC). It was found that FAA-DOPO could be used as a monomer for polymerization, although FAA-DOPO had a large steric hindrance from the chemical structure of FAA-DOPO, the UL-94 grade of PS-b-PFAA-DOPO reached the V-0 grade, and the LOI increased by 59.12% compared with PS148-Br.

Synthesis of High-Molecular-Weight Bifunctional Additives with both Flame Retardant Properties and Antistatic Properties via ATRP.

Polystyrene (PS) is widely used in our daily life, but it is flammable and produces a large number of toxic gases and high-temperature flue gases in the combustion process, which limit its application. Improving the flame retardancy of PS has become an urgent problem to be solved. In addition, in view of the disadvantage that small-molecule flame retardants can easily migrate from polymers during use, which leads to the gradual reduction of the flame retardant effect or even loss of flame retardant performance, and the outstanding advantages of ATRP technology in polymer structure design and function customization, we used ATRP technology to synthesize the high-molecular-weight bifunctional additive PFAA-DOPO-b-PDEAEMA, which has flame retardant properties and antistatic properties. The chemical structure and molecular weight of PFAA-DOPO-b-PDEAEMA were characterized by FTIR, 1H NMR, GPC, and XPS. When the addition of PFAA-DOPO-b-PDEAEMA was 15 wt %, the limiting oxygen index (LOI) of polystyrene composites was 28.4%, which was 53.51% higher than that of pure polystyrene, the peak of the heat release rate (pHRR) was 37.61% lower than that of pure polystyrene, UL-94 reached V-0 grade, and the flame retardant index (FRI) was 2.98. In addition, when the PFAA-DOPO-b-PDEEMA content is 15 wt %, the surface resistivity and volume resistivity of polystyrene composites are 2 orders of magnitude lower than those of polystyrene. This research work provides a reference for the design of bifunctional and even multifunctional polymers.

Research on Microstructure and Mechanical Properties of Nylon6/Basalt Fiber/High-Density Polyethylene Composites.

As a representative polyolefin, high-density polyethylene (HDPE) has become one of the most commonly used commercial plastics with a wide range of applications in the world. However, its applications are limited due to poor mechanical properties. Hence, it is indispensable to develop composites with improved mechanical properties to overcome this disadvantage. In our work, basalt fiber (BF) and polyamide 6 (PA6)-reinforced HDPE composites were prepared. The effects of adding fiber, organic filler, and polar component maleic anhydride (MAH) on the microstructural characteristics of composites were investigated. Microstructural characterization evidenced that the binary-dispersed phase (PA6/BF) possesses a core-shell structure in which the component PA6 encapsulates the component BF, and the extent of encapsulation declines with the increase of MAH addition. It has been confirmed by scanning electron microscopy (SEM) observation that the microstructure is related to the interfacial tension of components. The effects of multicomponents on the crystallization behavior of composites were studied. The differential scanning calorimeter (DSC) analysis exhibited a significant change in the HDPE microstructure. Results showed that, as nucleating agents, PA6 and BF improve the crystallization rate in the cooling process. Furthermore, the rheological behavior of multicomponent composites was studied. With the increase of MAH, a clear improvement of complex viscosity and storage modulus was observed, of which the mechanism has been discussed in detail. The relationship between microstructure and heat resistance of composites was studied by a thermal deformation test under static fore. It is confirmed that the thermally conductive fiber BF and other components can form a thermally conductive network and channels, thus improving the heat resistance. It can become a composite material, which is suitable for special environments.

Dual-Responsive Curcumin-Loaded Nanoparticles for the Treatment of Cisplatin-Induced Acute Kidney Injury.

Acute kidney injury (AKI) has been a global public health concern leading to high patient morbidity and mortality in the world. Nanotechnology-mediated antioxidative therapy has facilitated the treatment of AKI. Herein, a hierarchical curcumin-loaded nanodrug delivery system (NPS@Cur) was fabricated for antioxidant therapy to ameliorate AKI. The nanoplatform could respond to subacidic and reactive oxygen species (ROS) microenvironments. The subacidic microenvironment led to a smaller size (from 140.9 to 99.36 nm) and positive charge (from -4.9 to 12.6 mV), contributing to the high accumulation of nanoparticles. An excessive ROS microenvironment led to nanoparticle degradation and drug release. In vitro assays showed that NPS@Cur could scavenge excessive ROS and relieve oxidative stress in H2O2-induced HK-2 cells through reduced apoptosis, activated autophagy, and decreased endoplasmic reticulum stress. Results from cisplatin-induced AKI models revealed that NPS@Cur could effectively alleviate mitochondria injury and protect kidneys via antioxidative protection, activated autophagy, decreased endoplasmic reticulum stress, and reduced apoptosis. NPS@Cur showed excellent biocompatibility and low toxicity to primary tissues in mice. These results revealed that NPS@Cur may be a potential therapeutic strategy for efficiently treating cisplatin or other cause-induced AKI.

WAPO-A1 is the causal gene of the 7AL QTL for spikelet number per spike in wheat.

Improving our understanding of the genes regulating grain yield can contribute to the development of more productive wheat varieties. Previously, a highly significant QTL affecting spikelet number per spike (SNS), grain number per spike (GNS) and grain yield was detected on chromosome arm 7AL in multiple genome-wide association studies. Using a high-resolution genetic map, we established that the A-genome homeolog of WHEAT ORTHOLOG OF APO1 (WAPO-A1) was a leading candidate gene for this QTL. Using mutants and transgenic plants, we demonstrate in this study that WAPO-A1 is the causal gene underpinning this QTL. Loss-of-function mutants wapo-A1 and wapo-B1 showed reduced SNS in tetraploid wheat, and the effect was exacerbated in wapo1 combining both mutations. By contrast, spikes of transgenic wheat plants carrying extra copies of WAPO-A1 driven by its native promoter had higher SNS, a more compact spike apical region and a smaller terminal spikelet than the wild type. Taken together, these results indicate that WAPO1 affects SNS by regulating the timing of terminal spikelet formation. Both transgenic and wapo1 mutant plants showed a wide range of floral abnormalities, indicating additional roles of WAPO1 on wheat floral development. Previously, we found three widespread haplotypes in the QTL region (H1, H2 and H3), each associated with particular WAPO-A1 alleles. Results from this and our previous study show that the WAPO-A1 allele in the H1 haplotype (115-bp deletion in the promoter) is expressed at significantly lower levels in the developing spikes than the alleles in the H2 and H3 haplotypes, resulting in reduced SNS. Field experiments also showed that the H2 haplotype is associated with the strongest effects in increasing SNS and GNS (H2>H3>H1). The H2 haplotype is already present in most modern common wheat varieties but is rare in durum wheat, where it might be particularly useful to improve grain yield.

Stress granule-associated TaMBF1c confers thermotolerance through regulating specific mRNA translation in wheat (Triticum aestivum).

Heat stress is a major limiting factor for global wheat production and causes dramatic yield loss worldwide. The TaMBF1c gene is upregulated in response to heat stress in wheat. Understanding the molecular mechanisms associated with heat stress responses will pave the way to improve wheat thermotolerance. Through CRISPR/Cas9-based gene editing, polysome profiling coupled with RNA-sequencing analysis, and protein-protein interactions, we show that TaMBF1c conferred heat response via regulating a specific gene translation in wheat. The results showed that TaMBF1c is evolutionarily conserved in diploid, tetraploid and hexaploid wheat species, and its knockdown and knockout lines show increased heat sensitivity. TaMBF1c is colocalized with the stress granule complex and interacts with TaG3BP. TaMBF1c affects the translation efficiency of a subset of heat responsive genes, which are significantly enriched in the 'sequence-specific DNA binding' term. Moreover, gene expression network analysis demonstrated that TaMBF1c is closely associated with the translation of heat shock proteins. Our findings reveal a contribution of TaMBF1c in regulating the heat stress response via the translation process, and provide a new target for improving heat tolerance in wheat breeding programs.

A C-terminal encoded peptide, ZmCEP1, is essential for kernel development in maize.

C-terminal encoded peptides (CEPs) are peptide hormones which act as mobile signals coordinating important developmental programs. Previous studies have unraveled that CEPs are able to regulate plant growth and abiotic stress via cell-to-cell communication in Arabidopsis and rice; however, little is known about their roles in maize. Here, we examined the spatiotemporal expression pattern of ZmCEP1 and showed that ZmCEP1 is highly expressed in young ears and tassels of maize, particularly in the vascular bundles of ears. Heterologous expression of ZmCEP1 in Arabidopsis results in smaller plants and seed size. Similarly, overexpression of ZmCEP1 in maize decreased the plant and ear height, ear length, kernel size, and 100-kernel weight. Consistently, exogenous application of the synthesized ZmCEP1 peptide to the roots of Arabidopsis and maize inhibited root elongation. Knock-out of ZmCEP1 through CRISPR/Cas9 significantly increased plant and ear height, kernel size and 100-kernel weight. Transcriptome analysis revealed that knock-out of ZmCEP1 up-regulated a subset of genes involved in nitrogen metabolism, nitrate transport, sugar transport and auxin response. Thus, these results provide new insights into the genetic and molecular function of ZmCEP1 in regulating kernel development and plant growth, providing novel opportunities for maize breeding.