Using a New Fe3O4@CPAM Magnetic Flocculant and Microwave to Demulsify Heavy Oil-in-Water Emulsions.

In this study, cationic polyacrylamide (CPAM)-coated magnetic nanoparticles (MNPs) Fe3O4@CPAM were synthesized for treating heavy O/W emulsions. This Fe3O4@CPAM was characterized by Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), and vibrating sample magnetometry (VSM) techniques, and its synergistic performances with microwaves were evaluated in detail with respect to the microwave radiation power, radiation time, and magnetic nanoparticle concentration. On this basis, the distribution of oil droplets and the wettability and chargeability of magnetic nanoparticles were measured without or with microwave radiation using biomicroscopy, contact angle measurement instrument, and a ζ-potential analyzer, thus revealing the synergistic demulsification mechanism between microwave and magnetic nanoparticles. The results showed that excessively high or low microwave radiation parameters had an inhibitory effect on the magnetic nanoparticle demulsification, and microwave promoted the magnetic nanoparticle demulsification only when the radiation parameters were in the optimal range. In addition, the water separation rate showed an increasing and then decreasing trend with the increase of magnetic nanoparticles concentration, with or without microwave action. As an example, the water separation rate of the emulsion for 1 h was 21.34% when the Fe3O4 concentration was 175 mg/L without microwave action, while it increased to 55.56% with microwave action. In contrast, when the concentration of Fe3O4@CPAM was 175 mg/L, the water separation rate was 42.86% without microwave radiation, while it was further increased to 77.38% under microwave radiation. These results indicate that magnetic nanoparticles and their complexes significantly affect the water separation process under different conditions. There is a more obvious coupling synergistic effect between Fe3O4@CPAM and microwave. This was due to the lower absolute potential of Fe3O4@CPAM and its higher hydrophobicity.

N6-methyladenosine-mediated feedback regulation of abscisic acid perception via phase-separated ECT8 condensates in Arabidopsis.

N6-methyladenosine (m6A) is the most abundant internal modification in eukaryotic mRNAs, yet how plants recognize this chemical modification to swiftly adjust developmental plasticity under environmental stresses remains unclear. Here we show that m6A mRNA modification and its reader protein EVOLUTIONARILY CONSERVED C-TERMINAL REGION 8 (ECT8) act together as a key checkpoint for negative feedback regulation of abscisic acid (ABA) signalling by sequestering the m6A-modified ABA receptor gene PYRABACTIN RESISTANCE 1-LIKE 7 (PYL7) via phase-separated ECT8 condensates in stress granules in response to ABA. This partially depletes PYL7 mRNA from its translation in the cytoplasm, thus reducing PYL7 protein levels and compromising ABA perception. The loss of ECT8 results in defective sequestration of m6A-modified PYL7 in stress granules and permits more PYL7 transcripts for translation. This causes overactivation of ABA-responsive genes and the consequent ABA-hypersensitive phenotypes, including drought tolerance. Overall, our findings reveal that m6A-mediated sequestration of PYL7 by ECT8 in stress granules negatively regulates ABA perception, thereby enabling prompt feedback regulation of ABA signalling to prevent plant cell overreaction to environmental stresses.

Arnicolide D induces endoplasmic reticulum stress-mediated oncosis via ATF4 and CHOP in hepatocellular carcinoma cells.

Endoplasmic reticulum (ER) stress can trigger various cell death mechanisms beyond apoptosis, providing promise in cancer treatment. Oncosis, characterized by cellular swelling and increased membrane permeability, represents a non-apoptotic form of cell death. In our study, we discovered that Arnicolide D (AD), a natural sesquiterpene lactone compound, induces ER stress-mediated oncosis in hepatocellular carcinoma (HCC) cells, and this process is reactive oxygen species (ROS)-dependent. Furthermore, we identified the activation of the PERK-eIF2α-ATF4-CHOP pathway during ER stress as a pivotal factor in AD-induced oncosis. Notably, the protein synthesis inhibitor cycloheximide (CHX) was found to effectively reverse AD-induced oncosis, suggesting ATF4 and CHOP may hold crucial roles in the induction of oncosis by AD. These proteins play a vital part in promoting protein synthesis during ER stress, ultimately leading to cell death. Subsequent studies, in where we individually or simultaneously knocked down ATF4 and CHOP in HCC cells, provided further confirmation of their indispensable roles in AD-induced oncosis. Moreover, additional animal experiments not only substantiated AD's ability to inhibit HCC tumor growth but also solidified the essential role of ER stress-mediated and ROS-dependent oncosis in AD's therapeutic potential. In summary, our research findings strongly indicate that AD holds promise as a therapeutic agent for HCC by its ability to induce oncosis.

Plant RNA-binding proteins: Phase separation dynamics and functional mechanisms underlying plant development and stress responses.

RNA-binding proteins (RBPs) accompany RNA from synthesis to decay, mediating every aspect of RNA metabolism and impacting diverse cellular and developmental processes in eukaryotes. Many RBPs undergo phase separation along with their bound RNA to form and function in dynamic membraneless biomolecular condensates for spatiotemporal coordination or regulation of RNA metabolism. Increasing evidence suggests that phase-separating RBPs with RNA-binding domains and intrinsically disordered regions play important roles in plant development and stress adaptation. Here, we summarize the current knowledge about how dynamic partitioning of RBPs into condensates controls plant development and enables sensing of experimental changes to confer growth plasticity under stress conditions, with a focus on the dynamics and functional mechanisms of RBP-rich nuclear condensates and cytoplasmic granules in mediating RNA metabolism. We also discuss roles of multiple factors, such as environmental signals, protein modifications, and N6-methyladenosine RNA methylation, in modulating the phase separation behaviors of RBPs, and highlight the prospects and challenges for future research on phase-separating RBPs in crops.

A dephosphorylation-dependent molecular switch for FT repression mediates flowering in Arabidopsis.

The reproductive success of flowering plants relies greatly on precise timing of the floral transition, which is finely modulated by a complex network of floral regulators. As a main floral integrator, FLOWERING LOCUS T (FT) is also an essential constituent of the florigen that is transported from leaves to shoot apices to induce flowering. FT is specifically transcribed in leaf vascular tissues, where its production is suppressed by many flowering repressors, including the MYB transcription factor EARLY FLOWERING MYB PROTEIN (EFM). Here, we show that a plant CTD phosphatase, C-TERMINAL DOMAIN PHOSPHATASE-LIKE 2 (CPL2), suppresses FT expression in leaf vascular tissues by modulating the binding activity of EFM. CPL2 interacts with and dephosphorylates EFM to facilitate the binding of dephosphorylated EFM to FT chromatin, thereby inhibiting flowering. Our results suggest that CPL2-mediated dephosphorylation of the floral repressor EFM serves as a molecular switch, adding another layer of regulation to fine-tune FT transcription and ensure that flowering occurs at an appropriate time.

In Vitro and In Vivo Anti-Cancer Activity of Lasiokaurin in a Triple-Negative Breast Cancer Model.

Due to its intricate heterogeneity, high invasiveness, and poor prognosis, triple-negative breast cancer (TNBC) stands out as the most formidable subtype of breast cancer. At present, chemotherapy remains the prevailing treatment modality for TNBC, primarily due to its lack of estrogen receptors (ERs), progesterone receptors (PRs), and human epidermal growth receptor 2 (HER2). However, clinical chemotherapy for TNBC is marked by its limited efficacy and a pronounced incidence of adverse effects. Consequently, there is a pressing need for novel drugs to treat TNBC. Given the rich repository of diverse natural compounds in traditional Chinese medicine, identifying potential anti-TNBC agents is a viable strategy. This study investigated lasiokaurin (LAS), a natural diterpenoid abundantly present in Isodon plants, revealing its significant anti-TNBC activity both in vitro and in vivo. Notably, LAS treatment induced cell cycle arrest, apoptosis, and DNA damage in TNBC cells, while concurrently inhibiting cell metastasis. In addition, LAS effectively inhibited the activation of the phosphatidylinositol-3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/Akt/mTOR) pathway and signal transducer and activator of transcription 3 (STAT3), thus establishing its potential for multitarget therapy against TNBC. Furthermore, LAS demonstrated its ability to reduce tumor growth in a xenograft mouse model without exerting detrimental effects on the body weight or vital organs, confirming its safe applicability for TNBC treatment. Overall, this study shows that LAS is a potent candidate for treating TNBC.

Pan-centromere reveals widespread centromere repositioning of soybean genomes.

Centromere repositioning refers to a de novo centromere formation at another chromosomal position without sequence rearrangement. This phenomenon was frequently encountered in both mammalian and plant species and has been implicated in genome evolution and speciation. To understand the dynamic of centromeres on soybean genome, we performed the pan-centromere analysis using CENH3-ChIP-seq data from 27 soybean accessions, including 3 wild soybeans, 9 landraces, and 15 cultivars. Building upon the previous discovery of three centromere satellites in soybean, we have identified two additional centromere satellites that specifically associate with chromosome 1. These satellites reveal significant rearrangements in the centromere structures of chromosome 1 across different accessions, consequently impacting the localization of CENH3. By comparative analysis, we reported a high frequency of centromere repositioning on 14 out of 20 chromosomes. Most newly emerging centromeres formed in close proximity to the native centromeres and some newly emerging centromeres were apparently shared in distantly related accessions, suggesting their emergence is independent. Furthermore, we crossed two accessions with mismatched centromeres to investigate how centromere positions would be influenced in hybrid genetic backgrounds. We found that a significant proportion of centromeres in the S9 generation undergo changes in size and position compared to their parental counterparts. Centromeres preferred to locate at satellites to maintain a stable state, highlighting a significant role of centromere satellites in centromere organization. Taken together, these results revealed extensive centromere repositioning in soybean genome and highlighted how important centromere satellites are in constraining centromere positions and supporting centromere function.

Anti-osteoporosis bioactivity evaluation in zebrafish model of raw and salt-processed Achyranthes bidentata followed by liquid chromatography-mass spectrometry analysis and correlation analysis.

Achyranthes bidentata has been found to possess beneficial effects against osteoporosis, but there is still a lack of comprehensive studies on its anti-osteoporotic compounds. Therefore, in this study, we established a zebrafish osteoporosis model to evaluate the anti-osteoporotic effect of different fractions of raw and salt-processed A. bidentata. Among these fractions, the dichloromethane fraction showed the most promising anti-osteoporotic effect. To further investigate the active compounds responsible for the anti-osteoporosis effects, we prepared and analyzed the dichloromethane fraction of 10 batches of raw and salt-processed A. bidentata using liquid chromatography-mass spectrometry. As a result, we tentatively identified 19 compounds, including 11 saponins, three phenolic amides, three unsaturated fatty acids and two other compounds. To further narrow down the potential active compounds, we employed both orthogonal partial least squares discriminant analysis and gray relationship analysis. Through these analyses, we were able to identify eight compounds that showed a high correlation with the anti-osteoporosis effects of the dichloromethane fraction. Furthermore, we validated the anti-osteoporotic effects of ß-ecdysterone, wogonin, ginsenoside Ro, oleanolic acid, linoleic acid and palmitic acid using the zebrafish model. These compounds demonstrated significant anti-osteoporotic effects, further supporting their potential as active compounds in A. bidentata.

An in vitro Assay to Probe the Formation of Biomolecular Condensates.

Biomolecular condensates are membrane-less assemblies of proteins and nucleic acids formed through liquid-liquid phase separation (LLPS). These assemblies are known to temporally and spatially regulate numerous biological activities and cellular processes in plants and animals. In vitro phase separation assay using recombinant proteins represents one of the standard ways to examine the properties of proteins undergoing LLPS. Here, we present a detailed protocol to investigate in vitro LLPS using in vitro expressed and purified recombinant proteins.

MED8 regulates floral transition in Arabidopsis by interacting with FPA.

Success in plant reproduction is highly dependent on the correct timing of the floral transition, which is tightly regulated by the flowering pathways. In the model plant Arabidopsis thaliana, the central flowering repressor FLOWERING LOCUS C (FLC) is precisely regulated by multiple flowering time regulators in the vernalization pathway and autonomous pathway, including FPA. Here we report that Arabidopsis MEDIATOR SUBUNIT 8 (MED8) promotes floral transition in Arabidopsis by recruiting FPA to the FLC locus to repress FLC expression. Loss of MED8 function leads to a significant late-flowering phenotype due to increased FLC expression. We further show that MED8 directly interacts with FPA in the nucleus and recruits FPA to the FLC locus. Moreover, MED8 is indispensable for FPA's function in controlling flowering time and regulating FLC expression. Our study thus reveals a flowering mechanism by which the Mediator subunit MED8 represses FLC expression by facilitating the binding of FPA to the FLC locus to ensure appropriate timing of flowering for reproductive success.

Periodic changes in chain lengths distribution parameters of wheat starch during endosperm development.

This study analyzed the molecular structure of developing wheat endosperm starch at different stages after anthesis (DAA) using chain length distribution analysis by size exclusion chromatography (SEC) and fluorophore-assisted carbohydrate electrophoresis. Our results revealed periodic changes in the content of both amylose and amylopectin fractions. Specifically, the content of amylose chains with a degree of polymerization (DP) > 100 significantly decreased from 5 to 10 DAA (28% to 21%) and from 15 to 20 DAA (29% to 26%), but increased between 10 and 15 DAA (21% to 29%) and 20 to 25 DAA (30.0% to 33%). Conversely, the content of short amylopectin chains with DP ≤ 32 showed the opposite trend. Interestingly, mRNA expression levels of key starch biosynthesis genes did not exhibit periodic changes. These findings contribute to our understanding of starch biosynthesis and provide important insights for the development of starch-based products.

Nuclear translocation of OsMADS25 facilitated by OsNAR2.1 in reponse to nitrate signals promotes rice root growth by targeting OsMADS27 and OsARF7.

Nitrate is an important nitrogen source and signaling molecule that regulates plant growth and development. Although several components of the nitrate signaling pathway have been identified, the detailed mechanisms are still unclear. Our previous results showed that OsMADS25 can regulate root development in response to nitrate signals, but the mechanism is still unknown. Here, we try to answer two key questions: how does OsMADS25 move from the cytoplasm to the nucleus, and what are the direct target genes activated by OsMADS25 to regulate root growth after it moves to the nucleus in response to nitrate? Our results demonstrated that OsMADS25 moves from the cytoplasm to the nucleus in the presence of nitrate in an OsNAR2.1-dependent manner. Chromatin immunoprecipitation sequencing, chromatin immunoprecipitation qPCR, yeast one-hybrid, and luciferase experiments showed that OsMADS25 directly activates the expression of OsMADS27 and OsARF7, which are reported to be associated with root growth. Finally, OsMADS25-RNAi lines, the Osnar2.1 mutant, and OsMADS25-RNAi Osnar2.1 lines exhibited significantly reduced root growth compared with the wild type in response to nitrate supply, and expression of OsMADS27 and OsARF7 was significantly suppressed in these lines. Collectively, these results reveal a new mechanism by which OsMADS25 interacts with OsNAR2.1. This interaction is required for nuclear accumulation of OsMADS25, which promotes OsMADS27 and OsARF7 expression and root growth in a nitrate-dependent manner.

Recent advances in the plant epitranscriptome.

Chemical modifications of RNAs, known as the epitranscriptome, are emerging as widespread regulatory mechanisms underlying gene regulation. The field of epitranscriptomics advances recently due to improved transcriptome-wide sequencing strategies for mapping RNA modifications and intensive characterization of writers, erasers, and readers that deposit, remove, and recognize RNA modifications, respectively. Herein, we review recent advances in characterizing plant epitranscriptome and its regulatory mechanisms in post-transcriptional gene regulation and diverse physiological processes, with main emphasis on N6-methyladenosine (m6A) and 5-methylcytosine (m5C). We also discuss the potential and challenges for utilization of epitranscriptome editing in crop improvement.

Functional interdependence of N6-methyladenosine methyltransferase complex subunits in Arabidopsis.

Addition of N6-methyladenosine (m6A), the most prevalent internal mRNA modification in eukaryotes, is catalyzed by an evolutionarily conserved m6A methyltransferase complex. In the model plant Arabidopsis thaliana, the m6A methyltransferase complex is composed of 2 core methyltransferases, mRNA adenosine methylase (MTA) and MTB, and several accessory subunits such as FK506-BINDING PROTEIN 12 KD INTERACTING PROTEIN 37KD (FIP37), VIRILIZER (VIR), and HAKAI. It is yet largely unknown whether these accessory subunits influence the functions of MTA and MTB. Herein, I reveal that FIP37 and VIR are indispensable for stabilizing the methyltransferases MTA and MTB, thus functioning as key subunits to maintain the functionality of the m6A methyltransferase complex. Furthermore, VIR affects FIP37 and HAKAI protein accumulation, while MTA and MTB mutually influence each other. In contrast, HAKAI has little effect on protein abundance or localization of MTA, MTB, and FIP37. These findings uncover unique functional interdependence at the post-translational level among individual components in the Arabidopsis m6A methyltransferase complex, suggesting that maintenance of protein homeostasis among various subunits of the m6A methyltransferase complex is essential for maintaining the protein stoichiometry required for the proper function of the m6A methyltransferase complex in m6A deposition in plants.

Zoonotic fungus Arthroderma multifidum causing chronic pulmonary infection.

A rare case of fungus Arthroderma multifidum infection occurred in a 63-year-old man. The patient had some risk factors, including occupational exposure, immunosuppressive state, and structural basis following pulmonary tuberculosis and pneumothorax surgery. The pathogen was repeatedly isolated from bronchoalveolar lavage fluid and identified by gene sequencing. It is the first report of human infection caused by A. multifidum. Whole genome sequencing and analysis of its genomic characterization are completed. The findings provide us with a key clinical insight that the combination of immune suppression and environmental exposure could create an ideal condition for zoonotic fungal infections.

Shaping the landscape of N6-methyladenosine RNA methylation in Arabidopsis.

N 6-methyladenosine (m6A) modification on messenger RNAs (mRNAs) is deposited by evolutionarily conserved methyltransferases (writers). How individual m6A writers sculpt the overall landscape of the m6A methylome and the resulting biological impact in multicellular organisms remains unknown. Here, we systematically surveyed the quantitative m6A methylomes at single-nucleotide resolution and their corresponding transcriptomes in Arabidopsis (Arabidopsis thaliana) bearing respective impaired m6A writers. The m6A sites associated with the five Arabidopsis writers were located mostly within 3' untranslated regions with peaks at around 100 bp downstream of stop codons. m6A predominantly promoted the usage of distal poly(A) sites but had little effect on RNA splicing. Notably, impaired m6A writers resulted in hypomethylation and downregulation of transcripts encoding ribosomal proteins, indicating a possible correlation between m6A and protein translation. Besides the common effects on mRNA metabolism and biological functions uniquely exerted by different Arabidopsis m6A writers compared with their counterparts in human cell lines, our analyses also revealed the functional specificity of individual Arabidopsis m6A writers in plant development and response to stresses. Our findings thus reveal insights into the biological roles of various Arabidopsis m6A writers and their cognate counterparts in other multicellular m6A methyltransferase complexes.

NIN-at the heart of NItrogen-fixing Nodule symbiosis.

Legumes and actinorhizal plants establish symbiotic relationships with nitrogen-fixing bacteria, resulting in the formation of nodules. Nodules create an ideal environment for nitrogenase to convert atmospheric nitrogen into biological available ammonia. NODULE INCEPTION (NIN) is an indispensable transcription factor for all aspects of nodule symbiosis. Moreover, NIN is consistently lost in non-nodulating species over evolutions. Here we focus on recent advances in the signaling mechanisms of NIN during nodulation and discuss the role of NIN in the evolution of nitrogen-fixing nodule symbiosis.

The lead and cadmium content in rice and risk to human health in China: A systematic review and meta-analysis.

Numerous studies have investigated concentrations of lead (Pb) and cadmium (Cd) in rice in China, but have come to divergent conclusions. Therefore we systematically reviewed and meta-analyzed the available evidence on levels of Pb and Cd in rice in different regions of China in order to assess the potential risk to human health. The meta-analysis included 24 studies of Pb levels and 29 studies of Cd levels, published in 2011-2021. The pooled Pb concentration in rice was 0.10 mg per kg dry weight (95% CI 0.08-0.11), while the pooled Cd concentration was 0.16 mg per kg dry weight (95% CI 0.14-0.18). These levels are within the limits specified by national food safety standards. However, the total target hazard quotient for both metals exceeded 1.0 for adults and children, suggesting that rice consumption poses a health risk.

Experimental Study on Synergistic Demulsification of Microwave-Magnetic Nanoparticles.

Owing to the difficulty in the demulsification of heavy oil-in-water (O/W) emulsions, the demulsification rules of magnetic nanoparticles, microwave radiation, and magnetic-nanoparticle-assisted microwaves were investigated in this study. The surface potential and droplet size of the emulsion under different demulsification conditions were investigated by using a ζ potentiometer and polarizing microscopy to reveal the mechanism of demulsification. The results showed that γ-Fe2O3 exhibited the best demulsification performance among the six magnetic nanoparticles used for demulsification. With an increase in the concentration of γ-Fe2O3, the water separation of the heavy O/W emulsion first increased and then decreased, and with a decrease in pH, the demulsification performance gradually increased. The experimental results showed that microwave demulsification had an optimal power. The demulsification efficiency was significantly improved at the synergistic action between magnetic nanoparticles and the microwave, proving that magnetic nanoparticles had a promoting effect on microwave demulsification. In addition, the recycling experiment results showed that the magnetic nanoparticles exhibited good recyclability and reusability. Finally, a temperature field model of the emulsion under the synergistic effect of microwaves and magnetic nanoparticles was established and evaluated. Both before and after the addition of the magnetic nanoparticles, the theoretical temperature of the heavy O/W emulsion was consistent with the experimental temperature at different microwave powers and radiation times.

Insight into the Evolution of Ordered Mesoporous sp2 Carbonaceous Material Derived from Self-Assembly of a Block Copolymer.

Block-copolymer-derived ordered mesoporous carbon (OMC) materials have great potential in many applications, such as adsorption, catalysis, and energy conversions; however, their formation process and the kinetic mechanism remain unclear. Herein, a N-doped OMC (N-OMC) with sp2-bonded C atoms is developed via self-assembly of the polystyrene-block-poly(4-vinyl pyridine) block copolymer. By correlating the external morphologies with the internal chemical states, the formation process can be concluded as follows: (1) pore evolution via polystyrene domain degradation and (2) regularization and graphitization of the residual carbon via the removal of sp3 C atoms. In addition, the thickness of the N-OMC shows a power function relationship with the spin-coating rate, and the N content can be incredibly increased up to 26.34 at. % in an NH3 carbonization atmosphere. With the as-prepared N-OMC as the support for loading of the pseudo-atomic-scale Pt (Pt/N-OMC), a high electrochemical active surface area value of 99.64 m2·g-1 and a half-wave potential (E1/2) of 0.850 VRHE are achieved, showing great potential in developing single-atom electrocatalysts.