Molecular mechanism of HNTX-I activates the intermediate-conductance Ca2+-activated K+ (IK) channels.

The IK channel, a potassium ion channel regulated by calcium ions and voltages in a bidirectional manner, has been implicated in a range of diseases. However, there are currently few compounds available that can target the IK channel with high potency and specificity. Hainantoxin-I (HNTX-I) is the first peptide activator of IK channel discovered so far, but its activity is not ideal, and the underlying mechanism interaction between HNTX-I toxin and IK channel remains unclear. Thus, our study aimed to enhance the potency of IK channel activating peptides derived from HNTX-I and elucidate the molecular mechanism underlying the interaction between HNTX-I and the IK channel. By employing virtual alanine scanning mutagenesis, we generated 11 HNTX-I mutants using site-directed mutagenesis to pinpoint specific residues crucial for the HNTX-I and IK channel interaction. Subsequently, we identified key residues on the IK channel that are involved in the interaction with HNTX-I. Additionally, molecular docking was employed to guide the molecular engineering process and clarify the binding interface between HNTX-I and the IK channel. Our results demonstrate that HNTX-I primarily acts on the IK channel via the N-terminal amino acid, and its interaction with the IK channel is mediated by electrostatic and hydrophobic interactions, specifically the amino acid residues at positions 1, 3, 5, and 7 on HNTX-I. This study provides valuable insights into the peptide toxins that may serve as potential templates for the development of activators with enhanced potency and selectivity for the IK channel.

Modulation of anthocyanin accumulation in storage roots of sweetpotato by transcription factor IbMYB1-2 through direct binding to anthocyanin biosynthetic gene promoters.

The storage roots of purple-fleshed sweetpotato rich in anthocyanins are considered nutrient-rich foods with health effects. However, the molecular mechanism underlying anthocyanin biosynthesis and regulation remains to be revealed. In this study, IbMYB1-2 was isolated from purple-fleshed sweetpotato "Xuzishu8". The phylogenetic and sequence analysis indicated that IbMYB1-2 belongs to the SG6 subfamily with a conserved bHLH motif. Subcellular localization analysis and transcriptional activity assay revealed that IbMYB1-2 is a key transcriptional activator and is specific to the nucleus. Agrobacterium rhizogenes-mediated overexpression of IbMYB1-2 in sweetpotato through in vivo root transgenic system led to an increase in anthocyanins in the root of sweetpotato. qRT-PCR and transcriptome analysis depicted that the transcript levels of IbMYB1-2, IbbHLH42, and eight structural genes that are associated with the synthesis of anthocyanin were upregulated in overexpressed IbMYB1-2 transgenic roots. Dual-luciferase reporter (DLR) assay and yeast one-hybrid (Y1H) assay demonstrated IbMYB1-2 binding to the promoter regions of IbbHLH42 and other anthocyanin biosynthetic genes, including IbCHS, IbCHI, IbF3H, IbDFR, IbANS, IbGSTF12, IbUGT78D2, and IbUF3GT. Moreover, IbbHLH42 was shown to be an active enhancer for the formation of MYB-bHLH-WD40 (MBW) complex, which strongly supports the promoter activities of the IbCHS, IbANS, IbUGT78D2, and IbGSTF12 genes to induce anthocyanin accumulation. Taken together, our findings not only revealed the underlying regulatory molecular mechanism of IbMYB1-2 for anthocyanin accumulation in the storage roots of sweetpotato but also uncovered a potential mechanism by which IbbHLH42 modulated anthocyanin biosynthesis through a positive feedback regulatory loop.

Realgar­induced KRAS mutation lung cancer cell death via KRAS/Raf/MAPK mediates ferroptosis.

KRAS is a biomarker for non­small cell lung cancer­targeted therapy, but there is currently no effective KRAS­targeting medication. Realgar is an impelling anticancer drug, however its significance in KRAS mutant lung cancer is uncertain. According to our findings, the IC50 of H23 (KRAS mutant) cells is 2.99 times lower than that of H1650 (non­KRAS mutant) cells. Flow cytometry and the Hoechst 33258 staining assay revealed that H1650 cells treated with 4 µg/ml realgar had an apoptotic rate of 8.2%, while H23 cells had a rate of 21.46%. Accordingly, realgar was more sensitive to KRAS mutant cells. Transcriptome sequencing test indicated that there were 481 different expression genes in H23 cells treated with realgar. In H23 cells treated with realgar, mitochondria shrank, inner membrane folding was disturbed, and mitochondrial membrane potential crushed. Realgar boosted intracellular Fe2+, reactive oxygen species, malondialdehyde and glutathione levels, which were all reversed by ferroptosis inhibitor Fer­1. Realgar decreased phosphorylated p­Raf, p­ERK1/2 and increased p­p38 and p­JNK, whereas only p­Raf was abolished by Fer­1. Raf inhibitor Sorafenib accelerated the realgar­induced ferroptosis. On H23 cells treated with realgar, the expression of GPX4, SCL7A11 decreased while ACSL4 expression increased; this effect could also be amplified by Sorafenib. In conclusion, the present study indicated that realgar may induce ferroptosis by regulating the Raf, and hence plays a role in anti­KRAS mutant lung cancer.

A sectoral-level analysis of the short- and long-term impacts of the COVID-19 pandemic on China's stock market volatility.

Sampling China's Shenyin & Wanguo Sectoral Indices for 28 industries and 3272 listed firms included in those indices, and using industry- and firm-level daily data up to December 31, 2020, this paper empirically examined the short- and long-run impacts of the COVID-19 pandemic on stock return volatility. The results of the event study and univariate graphic analysis suggested that the market volatilities of the 28 industries were affected by COVID-19 events at various levels and that the events increased the volatility continuously for up to 6 days. The results of the panel data regression models revealed that the COVID-19-related daily new confirmed cases, daily new deaths, and cumulative cured cases were associated with higher volatility for all industries, although the impact levels were small; the daily deaths impacted volatility more than confirmed and cured cases. Finally, positive and significant effects of firm-specific variables such as total assets, turnover ratio and trading volume were recorded, indicating that fundamental aspects of the company and investors' behaviour also made great sense.

Comparative Transcriptome Analysis Reveals Key Genes and Pathways Involved in Prickle Development in Eggplant.

Eggplant is one of the most important vegetables worldwide. Prickles on the leaves, stems and fruit calyxes of eggplant may cause difficulties during cultivation, harvesting and transportation, and therefore is an undesirable agronomic trait. However, limited knowledge about molecular mechanisms of prickle morphogenesis has hindered the genetic improvement of eggplant. In this study, we performed the phenotypic characterization and transcriptome analysis on prickly and prickleless eggplant genotypes to understand prickle development at the morphological and molecular levels. Morphological analysis revealed that eggplant prickles were multicellular, lignified and layered organs. Comparative transcriptome analysis identified key pathways and hub genes involved in the cell cycle as well as flavonoid biosynthetic, photosynthetic, and hormone metabolic processes during prickle development. Interestingly, genes associated with flavonoid biosynthesis were up-regulated in developing prickles, and genes associated with photosynthesis were down-regulated in developing and matured prickles. It was also noteworthy that several development-related transcription factors such as bHLH, C2H2, MYB, TCP and WRKY were specifically down- or up-regulated in developing prickles. Furthermore, four genes were found to be differentially expressed within the Pl locus interval. This study provides new insights into the regulatory molecular mechanisms underlying prickle morphogenesis in eggplant, and the genes identified might be exploited in breeding programs to develop prickleless eggplant cultivars.

Phylogenetic Analysis of the SQUAMOSA Promoter-Binding Protein-Like Genes in Four Ipomoea Species and Expression Profiling of the IbSPLs During Storage Root Development in Sweet Potato (Ipomoea batatas).

As a major plant-specific transcription factor family, SPL genes play a crucial role in plant growth, development, and stress tolerance. The SPL transcription factor family has been widely studied in various plant species; however, systematic studies on SPL genes in the genus Ipomoea are lacking. Here, we identified a total of 29, 27, 26, and 23 SPLs in Ipomoea batatas, Ipomoea trifida, Ipomoea triloba, and Ipomoea nil, respectively. Based on the phylogenetic analysis of SPL proteins from model plants, the Ipomoea SPLs were classified into eight clades, which included conserved gene structures, domain organizations and motif compositions. Moreover, segmental duplication, which is derived from the Ipomoea lineage-specific whole-genome triplication event, was speculated to have a predominant role in Ipomoea SPL expansion. Particularly, tandem duplication was primarily responsible for the expansion of SPL subclades IV-b and IV-c. Furthermore, 25 interspecific orthologous groups were identified in Ipomoea, rice, Arabidopsis, and tomato. These findings support the expansion of SPLs in Ipomoea genus, with most of the SPLs being evolutionarily conserved. Of the 105 Ipomoea SPLs, 69 were predicted to be the targets of miR156, with seven IbSPLs being further verified as targets using degradome-seq data. Using transcriptomic data from aboveground and underground sweet potato tissues, IbSPLs showed diverse expression patterns, including seven highly expressed IbSPLs in the underground tissues. Furthermore, the expression of 11 IbSPLs was validated using qRT-PCR, and two (IbSPL17/IbSPL28) showed significantly increased expression during root development. Additionally, the qRT-PCR analysis revealed that six IbSPLs were strongly induced in the roots under phytohormone treatments, particularly zeatin and abscisic acid. Finally, the transcriptomic data of storage roots from 88 sweet potato accessions were used for weighted gene co-expression network analysis, which revealed four IbSPLs (IbSPL16/IbSPL17/IbSPL21/IbSPL28) clusters with genes involved in "regulation of root morphogenesis," "cell division," "cytoskeleton organization," and "plant-type cell wall organization or biogenesis," indicating their potential role in storage root development. This study not only provides novel insights into the evolutionary and functional divergence of the SPLs in the genus Ipomoea but also lays a foundation for further elucidation of the potential functional roles of IbSPLs on storage root development.

Genome-wide analysis of expression quantitative trait loci (eQTLs) reveals the regulatory architecture of gene expression variation in the storage roots of sweet potato.

Dissecting the genetic regulation of gene expression is critical for understanding phenotypic variation and species evolution. However, our understanding of the transcriptional variability in sweet potato remains limited. Here, we analyzed two publicly available datasets to explore the landscape of transcriptomic variations and its genetic basis in the storage roots of sweet potato. The comprehensive analysis identified a total of 724,438 high-confidence single nucleotide polymorphisms (SNPs) and 26,026 expressed genes. Expression quantitative trait locus (eQTL) analysis revealed 4408 eQTLs regulating the expression of 3646 genes, including 2261 local eQTLs and 2147 distant eQTLs. Two distant eQTL hotspots were found with target genes significantly enriched in specific functional classifications. By combining the information from regulatory network analyses, eQTLs and association mapping, we found that IbMYB1-2 acts as a master regulator and is the major gene responsible for the activation of anthocyanin biosynthesis in the storage roots of sweet potato. Our study provides the first insight into the genetic architecture of genome-wide expression variation in sweet potato and can be used to investigate the potential effects of genetic variants on key agronomic traits in sweet potato.

Rapidly monitoring the quality of flavoring essence based on microwave-induced plasma ionization mass spectrometry and multivariate statistical analysis.

Microwave-induced plasma ionization mass spectrometry (MIPI-MS) combined with multivariate statistical analysis was first applied to rapidly monitor the quality of tobacco flavoring essence. A small isolation and reaction chamber was set up between the ion source and the injection port of mass spectrometer to effectively eliminate the interference of external environment in the process of analyzing samples. The improved experimental apparatus (MIPI-MS) could achieve online and high-throughput analysis, with minimal sample preparation steps. Further, two types of tobacco flavoring essences with the similar appearance and physicochemical parameters were employed to verify the usability of the promising method in the field of quality monitoring. Firstly, the mass spectral fingerprint of each essence was established by the improved MIPI-MS method within 2 min. Then, two multivariate statistical processes were carried out to analyze mass spectral data. The similarity results indicated that the thresholds of tobacco flavoring essences from different batches were 1.512 and 2.638, respectively. The first three principal components of the established PLS-DA described 93.6% of the total variability, and provided a visualized comparison for the two types of flavoring materials. Finally, the adulterated samples were successfully distinguished by employing the two multivariate statistical processes.

Identification of nicotianamine synthase genes in Triticum monococcum and their expression under different Fe and Zn concentrations.

In graminaceous plants, nicotianamine (NA) is an important component of metal acquisition. NA is synthesized from S-adenosyl-l-methionine (SAM) catalyzed by nicotianamine synthase (NAS). Here, eight Triticum monococcum NAS (TmNAS) genes were cloned and characterized. Amino acid sequence analysis showed that TmNAS genes had high sequence identity with those from Triticum aestivum, Zea mays, Oryza sativa and Hordeum vulgare. Phylogenetic analysis showed that NAS genes were classified into two distinct groups, e.g. group I and group II. Expression analysis demonstrated that two of the TmNAS genes in group II were highly expressed in shoot tissues, and the other six TmNAS genes in group I were expressed in root tissues. Further analysis indicated that root-specific TmNAS genes were up-regulated under conditions of Fe- or Zn-deficiency growth, while shoot-specific TmNAS genes were up-regulated under conditions of Fe- or Zn-sufficiency. These results help us understand the NAS genes in T. monococcum and provide novel genetic resources for improving Fe and Zn concentrations in common wheat.

Preliminary construction of integral analysis for characteristic components in complex matrices by in-house fabricated solid-phase microextraction fibers combined with gas chromatography-mass spectrometry.

Integral analysis plays an important role in study and quality control of substances with complex matrices in our daily life. As the preliminary construction of integral analysis of substances with complex matrices, developing a relatively comprehensive and sensitive methodology might offer more informative and reliable characteristic components. Flavoring mixtures belonging to the representatives of substances with complex matrices have now been widely used in various fields. To better study and control the quality of flavoring mixtures as additives in food industry, an in-house fabricated solid-phase microextraction (SPME) fiber was prepared based on sol-gel technology in this work. The active organic component of the fiber coating was multi-walled carbon nanotubes (MWCNTs) functionalized with hydroxyl-terminated polydimethyldiphenylsiloxane, which integrate the non-polar and polar chains of both materials. In this way, more sensitive extraction capability for a wider range of compounds can be obtained in comparison with commercial SPME fibers. Preliminarily integral analysis of three similar types of samples were realized by the optimized SPME-GC-MS method. With the obtained GC-MS data, a valid and well-fit model was established by partial least square discriminant analysis (PLS-DA) for classification of these samples (R2X=0.661, R2Y=0.996, Q2=0.986). The validity of the model (R2=0.266, Q2=-0.465) has also approved the potential to predict the "belongingness" of new samples. With the PLS-DA and SPSS method, further screening out the markers among three similar batches of samples may be helpful for monitoring and controlling the quality of the flavoring mixtures as additives in food industry. Conversely, the reliability and effectiveness of the GC-MS data has verified the comprehensive and efficient extraction performance of the in-house fabricated fiber.

Near Infrared Quantum Cutting of Tb3+-Yb3+ Co-Doped CeF3 Nanophosphors.

In this paper, Tb3+-Yb3+ Co-doped CeF3 nanophosphors were synthesized using the microwave-assisted heating hydrothermal method (M-H). The excitation and emission spectra of the samples at room temperature show that the samples absorb ultraviolet light from 250 nm to 280 nm, and emit light at 300 nm. This corresponds to the transitions from 5D to 4F of Ce3+, 480 nm, 540 nm, 583 nm, 620 nm which correspond to the transitions from 5D4 to 7F6,5,4,3 of Tb3+, 973 nm which corresponds to the transitions from 2F5/2-2F7/2 of Yb3+. In the emission spectra, it is clear that the emission intensity of Ce3+ and Tb3+ decreases, and Yb3+ increases with increasing Yb3+. This suggests that energy transfer from Ce3+ to Yb3+, and Ce3+ to Tb3+ to Yb3+ may occur. In the near infrared emission area, it is noted that a distinct emission centered at 973 nm was observed under 260 nm excitation. This is due to transitions among the different Stark levels of 2FJ(J=5/2,7/2) Yb3+ ions. This also suggests an energy transfer from Ce3+ ions to Tb3+ and then to Yb3+. The energy transfers from Tb3+-Yb3+ Co-doped CeF3 nanophosphors, which lead to intense NIR emissions at 900-1050 nm, match the energy of Si band gaps of Si-based solar cells. Therefore, these kinds of materials are promising candidates for applications that require modifying if solar spectrums and enhancement of conversion efficiency of Si-based solar cells.

Sodium Citrate Inhibits Endoplasmic Reticulum Stress in Rats with Adenine-Induced Chronic Renal Failure.

BACKGROUND/AIMS: Endoplasmic reticulum stress (ERS) is an important self-protective cellular response to harmful stimuli that contribute to various diseases, including chronic renal failure (CRF). Sodium citrate plays an important role in antioxidant and cellular immunity, but whether it improves ERS in CRF is unclear. METHODS: The rats were randomly divided into five groups: the control group, the sodium citrate control group, the model group, model rats with low dose sodium citrate (216 mg/kg), and model rats with a high dose of sodium citrate (746 mg/kg). The rats were euthanized at 6, 8, 12, and 16 weeks with their blood and renal tissue in detection. RESULTS: The increased concentrations of blood urea nitrogen and serum creatinine in the model group were significantly decreased by sodium citrate treatment. Hematoxylin-eosin and Masson staining showed that sodium citrate treatment apparently improved renal pathological changes in CRF rats. Western blot analysis showed that sodium citrate treatment decreased the protein levels of transforming growth factor-beta 1 and collagen type IV, which were increased in model rats. Moreover, immunohistochemical staining demonstrated that sodium citrate could effectively reduce the protein expression of glucose-regulated protein 78 kDa and CCAAT/enhancer-binding protein homologous protein in the model rats, which was consistent with western blot results. Additionally, the high dose of sodium citrate had a stronger protective effect in CRF rats than the low dose of sodium citrate. CONCLUSIONS: Sodium citrate has a protective effect on CRF through its effects on ERS.

Identification of wheat non-specific lipid transfer proteins involved in chilling tolerance.

KEY MESSAGE: Three TaLTPs were found to enhance chilling tolerance of transgenic Arabidopsis, which were characterized by analyzes of promoter-GUS activity, subcellular localization, chromosomal location and transcriptional profile. Non-specific lipid transfer proteins (nsLTP) are abundantly expressed in plants, however, their functions are still unclear. In this study, we primarily characterized the functions of 3 type I TaLTP genes that were localized on chromosomes 3A, 3B, and 5D, respectively. The transcripts of TaLTPIb.1 and TaLTPIb.5 were induced under chilling, wound, and drought conditions, while TaLTPId.1 was only up-regulated by dark treatment. All the 3 TaLTP genes could be stimulated by the in vitro treatment of salicylic acid, while TaLTPId.1 was also positively regulated by methyljasmonic acid. Furthermore, the promoter-reporter assay of TaLTPIb.1 in the transgenic brachypodium showed a typical epidermis-specific expression pattern of this gene cluster. When fused with EGFP, all the 3 proteins were shown to localize on the plasma membrane in transgenic tobacco, although a signal in chloroplasts was also observed for TaLTPId.1. Heterogeneous overexpression of each of the TaLTP genes in Arabidopsis resulted in longer root length compared with wild type plants under chilling condition. These results suggest that type I TaLTPs may have a conserved functionality in chilling tolerance by lipid permeation in the plasma membrane of epidermal cells. On the other hand, the type I TaLTPs may exert functional divergence mainly through regulatory subfunctionalization.

[Upconversion properties of sodium and aluminum fluorides coated BaF2 material].

Sodium and aluminum fluorides coated BaF2 was synthesized by hydrothermal method. It was characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and fluorescence spectrophotometer, respectively. The analysis results indicated that sodium and aluminum fluorides existed in the form of sodium fluoride and sodium fluoroaluminate, which combined with BaF2 surface via chemical bonds, and exhibited coated structure. Fluorescence spectroscopy testing showed that there were two wide upconversion emission peaks with maxima at 304 and 324 nm under the excitation at 583 and 863 nm, respectively. Its phosphorescence life time was greater than 18.4 ms, and the emission intensity exhibited a rising process from the beginning and a decaying process after 15 ms. The luminescence mechanism was proposed according to the energy conversion process of the upconversion emission, the results showed that the upconversion emission belonged to the quantum confinement effect-interface light emission center radiative recombination.