Preparation and Photodegradation of TiO2 Thin Films on the Inner Wall of Quartz Tubes.

Titanium dioxide thin films on the inner wall of quartz tubes were prepared in situ by the sol-gel method. Meanwhile, copper and cerium were loaded onto the surface of the titanium dioxide thin films to enhance photocatalytic activity and broaden the range of light absorption. X-ray diffractometer, X-ray photoelectron spectroscopy, scanning electron microscopy, energy dispersive X-ray spectrum, N2 gas adsorption, UV diffuse reflectance spectroscopy, electron paramagnetic resonance, photoluminescene spectroscopy, and so on were used to characterize the structure, morphology, chemical composition, and optical properties of the prepared photocatalyst. Methylene blue (MB) was used as a simulated organic pollutant to study the photocatalytic performance of the photocatalyst, which was a translucent, structurally stable, and reusable high-efficiency photocatalytic catalyst. Under UV lamp irradiation, the MB photodegradation efficiency was 94.5%, which reached 91.2% after multiple cycles.

Comprehensive mapping and modelling of the rice regulome landscape unveils the regulatory architecture underlying complex traits.

Unraveling the regulatory mechanisms that govern complex traits is pivotal for advancing crop improvement. Here we present a comprehensive regulome atlas for rice (Oryza sativa), charting the chromatin accessibility across 23 distinct tissues from three representative varieties. Our study uncovers 117,176 unique open chromatin regions (OCRs), accounting for ~15% of the rice genome, a notably higher proportion compared to previous reports in plants. Integrating RNA-seq data from matched tissues, we confidently predict 59,075 OCR-to-gene links, with enhancers constituting 69.54% of these associations, including many known enhancer-to-gene links. Leveraging this resource, we re-evaluate genome-wide association study results and discover a previously unknown function of OsbZIP06 in seed germination, which we subsequently confirm through experimental validation. We optimize deep learning models to decode regulatory grammar, achieving robust modeling of tissue-specific chromatin accessibility. This approach allows to predict cross-variety regulatory dynamics from genomic sequences, shedding light on the genetic underpinnings of cis-regulatory divergence and morphological disparities between varieties. Overall, our study establishes a foundational resource for rice functional genomics and precision molecular breeding, providing valuable insights into regulatory mechanisms governing complex traits.

MYC phase separation selectively modulates the transcriptome.

Dysregulation and enhanced expression of MYC transcription factors (TFs) including MYC and MYCN contribute to the majority of human cancers. For example, MYCN is amplified up to several hundredfold in high-risk neuroblastoma. The resulting overexpression of N-myc aberrantly activates genes that are not activated at low N-myc levels and drives cell proliferation. Whether increasing N-myc levels simply mediates binding to lower-affinity binding sites in the genome or fundamentally changes the activation process remains unclear. One such activation mechanism that could become important above threshold levels of N-myc is the formation of aberrant transcriptional condensates through phase separation. Phase separation has recently been linked to transcriptional regulation, but the extent to which it contributes to gene activation remains an open question. Here we characterized the phase behavior of N-myc and showed that it can form dynamic condensates that have transcriptional hallmarks. We tested the role of phase separation in N-myc-regulated transcription by using a chemogenetic tool that allowed us to compare non-phase-separated and phase-separated conditions at equivalent N-myc levels, both of which showed a strong impact on gene expression compared to no N-myc expression. Interestingly, we discovered that only a small percentage (<3%) of N-myc-regulated genes is further modulated by phase separation but that these events include the activation of key oncogenes and the repression of tumor suppressors. Indeed, phase separation increases cell proliferation, corroborating the biological effects of the transcriptional changes. However, our results also show that >97% of N-myc-regulated genes are not affected by N-myc phase separation, demonstrating that soluble complexes of TFs with the transcriptional machinery are sufficient to activate transcription.

Phase separation of YAP-MAML2 differentially regulates the transcriptome.

Phase separation (PS) drives the formation of biomolecular condensates that are emerging biological structures involved in diverse cellular processes. Recent studies have unveiled PS-induced formation of several transcriptional factor (TF) condensates that are transcriptionally active, but how strongly PS promotes gene activation remains unclear. Here, we show that the oncogenic TF fusion Yes-associated protein 1-Mastermind like transcriptional coactivator 2 (YAP-MAML2) undergoes PS and forms liquid-like condensates that bear the hallmarks of transcriptional activity. Furthermore, we examined the contribution of PS to YAP-MAML2-mediated gene expression by developing a chemogenetic tool that dissolves TF condensates, allowing us to compare phase-separated and non-phase-separated conditions at identical YAP-MAML2 protein levels. We found that a small fraction of YAP-MAML2-regulated genes is further affected by PS, which include the canonical YAP target genes CTGF and CYR61, and other oncogenes. On the other hand, majority of YAP-MAML2-regulated genes are not affected by PS, highlighting that transcription can be activated effectively by diffuse complexes of TFs with the transcriptional machinery. Our work opens new directions in understanding the role of PS in selective modulation of gene expression, suggesting differential roles of PS in biological processes.

Poly(ethylene succinate-co-lactic acid) as a Multifunctional Additive for Modulating the Miscibility, Crystallization, and Mechanical Properties of Poly(lactic acid).

Polymer blending offers an effective and economical approach to overcome the performance limitations of poly(lactic acid) (PLA). In this study, a series of copolymers poly(ethylene succinate-co-lactic acid) (PESL) were synthesized, featuring lactic acid (LA) contents that ranged from 20 to 86 wt %. This synthesis involved a one-pot industrial melt polycondensation process using succinic acid (SA), ethylene glycol (EG), and LA, catalyzed by titanium tetraisopropoxide (TTP). The goal was to produce a fully biobased copolymer expected to exhibit partial miscibility with pure poly(lactic acid) (PLA). To assess the capability of PESL copolymers in toughening PLA, we conducted tensile testing on PLA/PESL blends containing 15 wt % PESL. As a result, an elongation at break for the blends with 15 wt % loading of the copolymer PESL72 was directly enhanced to 250% with an ultimate strength of 35 MPa, compared to brittle PLA with less 10% tensile length. The morphological features of interfacial adhesion before and after tensile failure were measured by scanning electron microscopy (SEM). A significant enhancement in the chain mobility of the PLA/PESL blends was further evidenced by differential scanning calorimetry (DSC) and dynamic mechanical analysis (DMA). These findings hold promise for the development of functional packaging materials based on PLA. The proposed copolymer design, which boasts strong industrial feasibility, can serve as a valuable guide for enhancing the toughness of PLA.

Verification of Reinforced Surface Loose Layer of Zinc-Aluminum-Magnesium Steel Plate.

The corrosion resistance of zinc-aluminum-magnesium steel plates (Zn-Al-Mg steel plates) is significantly higher than that of galvanized steel plates. However, the unsatisfactory bonding performance of Zn-Al-Mg steel plates significantly limits their widespread application. In this study, X-ray photoelectron spectroscopy is employed to detect changes in the surface oxygen content of Zn-Al-Mg steel plates after different temperature treatments to confirm the existence of surface loose layers. In particular, changes in the surface oxygen content of the Zn-Al-Mg steel plates after the oxide layer is removed are investigated under saturated H2O vapor and O2 environmental conditions, and the cause of the formation of loose surface layers is determined. The uneven distribution of elements on the surface of the Zn-Al-Mg steel plates is investigated with scanning electron microscopy and energy dispersive spectroscopy. Nuclear magnetic resonance is employed to determine the size of the network spatial structure formed by silane coupling agents under different hydrolysis conditions and to further investigate the bonding performance of hydrolysate-modified Zn-Al-Mg steel plates. Several typical automotive adhesives are utilized to compare and examine the changes in the tensile strength of the Zn-Al-Mg steel plate bonding before and after modification with the silane coupling agent and analyze the structural damage of the adhesive at the bonding interface. The results confirm that the silane coupling agent strengthens the loose layer on the surface of the Zn-Al-Mg steel plate.

Chemogenetic Minitool for Dissecting the Roles of Protein Phase Separation.

Biomolecular condensate is an emerging structural entity that regulates various cellular processes. Recent studies have discovered the phase-separation (PS) capability of several transcription factors (TFs) including YAP/TAZ upon biological stimuli, which provide new mechanisms of gene regulation. However, it remains mostly unanswered as to whether PS from a diffuse state to a phase-separated state promotes gene transcription. To address this question, we have designed a chemogenetic tool, dubbed SPARK-ON, which manipulates the PS of YAP and TAZ without a biological stimulus, forming condensates that are transcriptionally active, containing the DNA-binding partner TEAD, genomic DNA, transcriptional machinery, and nascent RNA. Most importantly, PS of TAZ increases the transcription of its target genes. Therefore, our data indicate that PS promotes gene transcription of TAZ. SPARK-ON is advantageous to current mutagenesis-based approaches that are often problematic when mutagenesis affects the transcriptional activity of a TF. Furthermore, protein abundance levels also affect gene transcription, but PS depends on protein abundance because PS occurs only when the protein level is above a saturation concentration. SPARK-ON decouples PS from protein abundance levels without introducing mutations and thus will find important applications in understanding the biological roles of PS for many TFs and other biomolecular condensates.

ATM-SPARK: A GFP phase separation-based activity reporter of ATM.

The kinase ataxia telangiectasia mutated (ATM) plays a key role in the DNA damage response (DDR). It is thus essential to visualize spatiotemporal dynamics of ATM activity during DDR. Here, we designed a robust ATM activity reporter based on phosphorylation-inducible green fluorescent protein phase separation, dubbed ATM-SPARK (separation of phases-based activity reporter of kinase). Upon ATM activation, it undergoes phase separation via multivalent interactions, forming intensely bright droplets. The reporter visualizes spatiotemporal dynamics of endogenous ATM activity in living cells, and its signal is proportional to the amount of DNA damage. ATM-SPARK also enables high-throughput screening of biological and small-molecule regulators. We identified the protein phosphatase 4 that blocks ATM activity. We also identified BGT226 as a potent ATM inhibitor with a median inhibitory concentration of ~3.8 nanomolars. Furthermore, BGT226 sensitizes cancer cells to the radiomimetic drug neocarzinostatin, suggesting that BGT226 might be combined with radiotherapeutic treatment. ATM-SPARK achieves large dynamic range, bright fluorescence, and simple signal pattern.

Fluorogenic reporter enables identification of compounds that inhibit SARS-CoV-2.

The coronavirus SARS-CoV-2 causes the severe disease COVID-19. SARS-CoV-2 infection is initiated by interaction of the viral spike protein and host receptor angiotensin-converting enzyme 2 (ACE2). We report an improved bright and reversible fluorogenic reporter, named SURF (split UnaG-based reversible and fluorogenic protein-protein interaction reporter), that we apply to monitor real-time interactions between spike and ACE2 in living cells. SURF has a large dynamic range with a dark-to-bright fluorescence signal that requires no exogenous cofactors. Utilizing this reporter, we carried out a high-throughput screening of small-molecule libraries. We identified three natural compounds that block replication of SARS-CoV-2 in both Vero cells and human primary nasal and bronchial epithelial cells. Cell biological and biochemical experiments validated all three compounds and showed that they block the early stages of viral infection. Two of the inhibitors, bruceine A and gamabufotalin, were also found to block replication of the Delta and Omicron variants of SARS-CoV-2. Both bruceine A and gamabufotalin exhibited potent antiviral activity in K18-hACE2 and wild-type C57BL6/J mice, as evidenced by reduced viral titres in the lung and brain, and protection from alveolar and peribronchial inflammation in the lung, thereby limiting disease progression. We propose that our fluorescent assay can be applied to identify antiviral compounds with potential as therapeutic treatment for COVID-19 and other respiratory diseases.

Surface coating preparation of spherical magnetic materials.

Magnetic materials are being increasingly used in anti-counterfeiting coatings, but the dark colors of magnetic materials greatly limit their applications. This necessitates the development of light-colored magnetic materials. In this study, the heterogeneous precipitation method was used to deposit a layer of titanium dioxide (TiO2) on the surface of magnetic spherical metal particles, followed by the deposition of a layer of Ag by the reduction method, in order to achieve a light color. In the experiment, the particles were initially coated with a few tens of nanometers of TiO2 with a strong shading effect, followed by a further coating of Ag of the same thickness with a similar shading performance. Not only did this achieve a lighter color, but there was no reduction in the magnetic properties of the material after the application of the coating. Scanning electron microscopy (SEM), scanning electron microscopy and energy-dispersive spectroscopy (SEM-EDS), X-ray diffractometry (XRD), and other methods were used to study the changes in morphology and composition before and after the magnetic material was coated. A magnetic tester was used to study the changes in magnetic strength before and after the magnetic material was coated.

Ring-opening polymerization-induced self-assembly (ROPISA) of salicylic acid o-carboxyanhydride.

Here is the first report on polyester-based nanocarriers fabricated via the ring-opening polymerization-induced self-assembly (ROPISA) of salicylic acid o-carboxyanhydride (SAOCA). This ROPISA process affords well-defined diblock copolymers that interestingly form an original cylindrical morphology.

Ethacridine inhibits SARS-CoV-2 by inactivating viral particles.

The respiratory disease COVID-19 is caused by the coronavirus SARS-CoV-2. Here we report the discovery of ethacridine as a potent drug against SARS-CoV-2 (EC50 ~ 0.08 µM). Ethacridine was identified via high-throughput screening of an FDA-approved drug library in living cells using a fluorescence assay. Plaque assays, RT-PCR and immunofluorescence imaging at various stages of viral infection demonstrate that the main mode of action of ethacridine is through inactivation of viral particles, preventing their binding to the host cells. Consistently, ethacridine is effective in various cell types, including primary human nasal epithelial cells that are cultured in an air-liquid interface. Taken together, our work identifies a promising, potent, and new use of the old drug via a distinct mode of action for inhibiting SARS-CoV-2.

Wheat TaPUB1 protein mediates ABA response and seed development through ubiquitination.

Abscisic acid (ABA) is an important regulator of plant growth, development, and biotic and abiotic stress responses. Ubiquitination plays important roles in regulating ABA signaling. E3 ligase, a key member in ubiquitination, actively participates in the regulation of biosynthesis, de-repression, and activation of ABA response and degradation of signaling components. In this study, we found that that overexpression of wheat E3 ligase TaPUB1 decreased the sensitivity of wheat seedlings to ABA, whereas TaPUB1-RNA interference (TaPUB1-RNAi) lines increased wheat sensitivity to ABA during germination, root growth, and stomatal opening. TaPUB1 influenced the expression of several ABA-responsive genes, and also interacted with TaPYL4 and TaABI5, which are involved in ABA signal transduction, and promoted their degradation. Additionally, we observed that TaPUB1-OE lines resulted in lower single-split grain numbers, larger seed size, and higher thousand kernel weight, when compared with the WT lines. Contrasting results were obtained for TaPUB1-RNAi lines. It suggests that TaPUB1 acts as a negative regulator in the ABA signaling pathway by interacting with TaPYL4 and TaABI5, subsequently affecting seed development in wheat. In addition, the enhanced abiotic tolerance of overexpression lines due to enhanced photosynthesis and root development may be related to the degradation of TaABI5 by TaPUB1.

Wheat TaPUB1 Regulates Cd Uptake and Tolerance by Promoting the Degradation of TaIRT1 and TaIAA17.

Cadmium (Cd) accumulation in agricultural soils is an increasingly serious problem, as plants absorb Cd, which inhibits their growth and development. Nonetheless, the molecular mechanisms underlying Cd detoxification and accumulation in wheat (Triticum aestivum L.) are unclear. Here, we isolated the U-box E3 ligase TaPUB1 from wheat and reported the functional characterization of TaPUB1 in Cd uptake and tolerance in wheat. Under Cd stress, TaPUB1 overexpression lines displayed higher photosynthetic rates than the wild type; opposite results were observed in the TaPUB1-RNAi lines. In addition, TaPUB1 overexpression lines showed reduced Cd uptake and accumulation, whereas RNAi plants exhibited a significant increase in Cd accumulation after Cd treatment. We further found that TaPUB1 enhanced the resistance of wheat to Cd stress in three ways. First, TaPUB1 interacts with and ubiquitinates TaIRT1, resulting in the inhibition of Cd uptake. Second, TaPUB1 interacts directly with and ubiquitinates TaIAA17, facilitates its degradation, and results in primary root elongation by activating the Aux signaling pathway under Cd stress. Moreover, TaPUB1 decreases ROS accumulation by regulating antioxidant-related gene expression and antioxidant enzyme activity under Cd stress. Thus, a molecular mechanism by which TaPUB1 regulates Cd uptake and tolerance by modulating the stability of TaIRT1 and TaIAA17 proteins was revealed.

Ethacridine inhibits SARS-CoV-2 by inactivating viral particles in cellular models.

SARS-CoV-2 is the coronavirus that causes the respiratory disease COVID-19, which is now the third-leading cause of death in the United States. The FDA has recently approved remdesivir, an inhibitor of SARS-CoV-2 replication, to treat COVID-19, though recent data from the WHO shows little to no benefit with use of this anti-viral agent. Here we report the discovery of ethacridine, a safe antiseptic use in humans, as a potent drug for use against SARS-CoV-2 (EC50 ~ 0.08 µM). Ethacridine was identified via high-throughput screening of an FDA-approved drug library in living cells using a fluorescent assay. Interestingly, the main mode of action of ethacridine is through inactivation of viral particles, preventing their binding to the host cells. Indeed, ethacridine is effective in various cell types, including primary human nasal epithelial cells. Taken together, these data identify a promising, potent, and new use of the old drug possessing a distinct mode of action for inhibiting SARS-CoV-2.

Expansin gene TaEXPA2 positively regulates drought tolerance in transgenic wheat (Triticum aestivum L.).

Expansins loosen plant cell walls and are involved in cell enlargement and various abiotic stresses. In previous studies, we cloned the expansin gene TaEXPA2 from the wheat cultivar HF9703. Here, we studied its function and regulation in wheat drought stress tolerance. The results indicated that TaEXPA2-overexpressing wheat plants (OE) exhibited drought tolerant phenotypes, whereas down-regulation of TaEXPA2 by RNA interference (RNAi) resulted in elevated drought sensitivity, as measured by survival rate, photosynthetic rate and water containing ability under drought stress. Overexpression of TaEXPA2 enhanced the antioxidant capacity in wheat plants, via elevation of antioxidant enzyme activity and the increase of the transcripts of some ROS scavenging enzyme-related genes. Further investigation revealed that TaEXPA2 positively influenced lateral root formation under drought conditions. A MYB transcription factor of wheat named TaMPS activates TaEXPA2 expression directly by binding to its promoter. Overexpression of TaMPS in Arabidopsis conferred drought tolerance associated with improved lateral root number, and the close homolog genes of TaEXPA2 were up-regulated in Arabidopsis roots overexpressing TaMPS, which suggest that TaMPS may function as one of the regulator of TaEXPA2 gene expression in the root lateral development under drought stress. These findings suggest that TaEXPA2 positively regulates drought stress tolerance in wheat.

The involvement of wheat U-box E3 ubiquitin ligase TaPUB1 in salt stress tolerance.

U-box E3 ubiquitin ligases play important roles in the ubiquitin/26S proteasome machinery and in abiotic stress responses. TaPUB1-overexpressing wheat (Triticum aestivum L.) were generated to evaluate its function in salt tolerance. These plants had more salt stress tolerance during seedling and flowering stages, whereas the TaPUB1-RNA interference (RNAi)-mediated knock-down transgenic wheat showed more salt stress sensitivity than the wild type (WT). TaPUB1 overexpression upregulated the expression of genes related to ion channels and increased the net root Na+ efflux, but decreased the net K+ efflux and H+ influx, thereby maintaining a low cytosolic Na+ /K+ ratio, compared with the WT. However, RNAi-mediated knock-down plants showed the opposite response to salt stress. TaPUB1 could induce the expression of some genes that improved the antioxidant capacity of plants under salt stress. TaPUB1 also interacted with TaMP (Triticum aestivum α-mannosidase protein), a regulator playing an important role in salt response in yeast and in plants. Thus, low cytosolic Na+ /K+ ratios and better antioxidant enzyme activities could be maintained in wheat with overexpression of TaPUB1 under salt stress. Therefore, we conclude that the U-box E3 ubiquitin ligase TaPUB1 positively regulates salt stress tolerance in wheat.

Wheat F-box Protein TaFBA1 Positively Regulates Plant Drought Tolerance but Negatively Regulates Stomatal Closure.

The phytohormone abscisic acid (ABA) regulates plant growth and development, as well as responses to various stresses, such as salt and drought. The wheat TaFBA1 gene, which encodes an F-box protein, was previously identified in our laboratory by homologous cloning. We previously found that TaFBA1 expression was induced by ABA and drought stress. In this study, wild-type (WT), TaFBA1 over-expressing (OEs), TaFBA1 homologous gene mutants, and TaFBA1 recovery (Rs) Arabidopsis plants were used. We found that the germination rate, the cotyledon greening rate, the root length, and the photosynthetic performance of TaFBA1 OE plants were better than those of WT under drought and ABA conditions, but mutant plants showed the opposite trend, and overexpression of TaFBA1 in mutants can recover their phenotype. In addition, TaFBA1 was found to be a negative regulator of ABA-induced stoma movement; mRNA transcription of certain ABA signaling-related genes was lower in TaFBA1 OE plants than in WT plants following ABA treatment. Further, we found that TaFBA1 can interact with RCAR1 (an ABA receptor) and ABI5. BiFC assay showed that TaFBA1 may interact with RCAR1 in the plasma membrane. In addition, accumulation of ROS and MDA in TaFBA1 OE plants was lower than that in the WT plants after ABA and drought treatments. Based on these results, we suggest that TaFBA1-regulated ABA insensitivity may be dependent on regulating ABA-mediated gene expression through interacting with RCAR1 and ABI5. Increased antioxidant competence and decreased ROS accumulation may be an important mechanism that underlies improved drought tolerance in TaFBA1 OE plants.

Designing a Green Fluorogenic Protease Reporter by Flipping a Beta Strand of GFP for Imaging Apoptosis in Animals.

A family of proteases called caspases mediate apoptosis signaling in animals. We report a GFP-based fluorogenic protease reporter, dubbed "FlipGFP", by flipping a beta strand of the GFP. Upon protease activation and cleavage, the beta strand is restored, leading to reconstitution of the GFP and fluorescence. FlipGFP-based TEV protease reporter achieves 100-fold fluorescence change. A FlipGFP-based executioner caspase reporter visualized apoptosis in live zebrafish embryos with spatiotemporal resolution. FlipGFP also visualized apoptotic cells in the midgut of Drosophila. Thus, the FlipGFP-based caspase reporter will be useful for monitoring apoptosis during animal development and for designing reporters of proteases beyond caspases. The design strategy can be further applied to a red fluorescent protein for engineering a red fluorogenic protease reporter.