Quaternity method for integrated screening, separation, extraction optimization, and bioactivity evaluation of acetylcholinesterase inhibitors from Sophora flavescens Aiton.

INTRODUCTION: Sophora flavescens Aiton (Fabaceae), a ubiquitous plant species in Asia, contains a wide range of pharmacologically active compounds, such as flavonoids, with potential anti-Alzheimer's disease (anti-AD) effects. OBJECTIVES: The objective of the study is to develop a quaternity method for the screening, isolation, extraction optimization, and activity evaluation of acetylcholinesterase (AChE)-inhibiting compounds from S. flavescens to realize high-throughput screening of active substances in traditional Chinese medicine and to provide experimental data for the development of anti-AD drugs. METHODS: With AChE as the target molecule, affinity ultrafiltration and liquid chromatography-mass spectrometry were applied to screen for potential inhibitors of the enzyme in S. flavescens. Orthogonal array experiments combined with the multi-objective Non-Dominated Sorting Genetic Algorithm III was used for the first time to optimize the process for extracting the active substances. Enzyme inhibition kinetics and molecular docking studies were performed to verify the potential anti-AD effects of the active compounds. RESULTS: Five AChE-inhibiting compounds were identified: kushenol I, kurarinone, sophoraflavanone G, isokurarinone, and kushenol E. These were successfully separated at purities of 72.88%, 98.55%, 96.86%, 96.74%, and 95.84%, respectively, using the n-hexane/ethyl acetate/methanol/water (4.0/5.0/4.0/5.0, v/v/v/v), n-hexane/ethyl acetate/methanol/water (5.0/5.0/6.0/4.0, v/v/v/v), and n-hexane/ethyl acetate/methanol/water (4.9/5.1/5.7/4.3, v/v/v/v) mobile phase systems. Enzyme inhibition kinetics revealed that kushenol E had the best inhibitory effect. CONCLUSION: This study elucidates the mechanism of action of five active AChE inhibitors in S. flavescens and provides a theoretical basis for the screening and development of anti-AD and other therapeutic drugs.

Insights into the tolerant function of VWA proteins in terms of expression analysis and RGLG5-VWA crystal structure.

The VWA domain commonly functions as a crucial component of multiprotein complexes, facilitating protein-protein interactions. However, limited studies have focused on the systemic study of VWA proteins in plants. Here, we identified 28 VWA protein genes in Arabidopsis thaliana, categorized into three clades, with one tandem duplication event and four paralogous genes within collinearity blocks. Then, we determined their expression patterns under abiotic stresses by transcriptomic analysis. All five RGLG genes were found to be responsive to at least one kind of abiotic stress, and RGLG5 was identified as a multiple stress-responsive gene, coding an E3 ubiquitin ligase with a VWA domain and a C-terminal RING domain. Subsequently, we explored tolerant function of RGLG5 by determining the crystal structure of its VWA domain. The structural comparison revealed the allosteric regulation mechanism of RGLG5-VWA, wherein the deflection of α7 led to displacement of key residue binding metal ion within MIDAS motif. Our findings provide full-scale knowledge on VWA proteins, and insights into tolerant function of RGLG5-VWA in terms of crystal structure.

Identification and Functional Analysis of KH Family Genes Associated with Salt Stress in Rice.

Salinity stress has a great impact on crop growth and productivity and is one of the major factors responsible for crop yield losses. The K-homologous (KH) family proteins play vital roles in regulating plant development and responding to abiotic stress in plants. However, the systematic characterization of the KH family in rice is still lacking. In this study, we performed genome-wide identification and functional analysis of KH family genes and identified a total of 31 KH genes in rice. According to the homologs of KH genes in Arabidopsis thaliana, we constructed a phylogenetic tree with 61 KH genes containing 31 KH genes in Oryza sativa and 30 KH genes in Arabidopsis thaliana and separated them into three major groups. In silico tissue expression analysis showed that the OsKH genes are constitutively expressed. The qRT-PCR results revealed that eight OsKH genes responded strongly to salt stresses, and OsKH12 exhibited the strongest decrease in expression level, which was selected for further study. We generated the Oskh12-knockout mutant via the CRISPR/Cas9 genome-editing method. Further stress treatment and biochemical assays confirmed that Oskh12 mutant was more salt-sensitive than Nip and the expression of several key salt-tolerant genes in Oskh12 was significantly reduced. Taken together, our results shed light on the understanding of the KH family and provide a theoretical basis for future abiotic stress studies in rice.

Genetic mapping reveals a candidate gene CmoFL1 controlling fruit length in pumpkin.

Fruit length (FL) is an important economical trait that affects fruit yield and appearance. Pumpkin (Cucurbita moschata Duch) contains a wealth genetic variation in fruit length. However, the natural variation underlying differences in pumpkin fruit length remains unclear. In this study, we constructed a F2 segregate population using KG1 producing long fruit and MBF producing short fruit as parents to identify the candidate gene for fruit length. By bulked segregant analysis (BSA-seq) and Kompetitive Allele-Specific PCR (KASP) approach of fine mapping, we obtained a 50.77 kb candidate region on chromosome 14 associated with the fruit length. Then, based on sequence variation, gene expression and promoter activity analyses, we identified a candidate gene (CmoFL1) encoding E3 ubiquitin ligase in this region may account for the variation of fruit length. One SNP variation in promoter of CmoFL1 changed the GT1CONSENSUS, and DUAL-LUC assay revealed that this variation significantly affected the promoter activity of CmoFL1. RNA-seq analysis indicated that CmoFL1 might associated with the cell division process and negatively regulate fruit length. Collectively, our work identifies an important allelic affecting fruit length, and provides a target gene manipulating fruit length in future pumpkin breeding.

Four-wavelength laser interferometry for the demodulation of dual-cavity fiber-optic extrinsic Fabry-Perot interferometric sensors.

A four-wavelength passive demodulation algorithm is proposed and experimentally demonstrated for the interrogation of the one cavity in a dual-cavity extrinsic Fabry-Perot interferometric (EFPI) sensor. The lengths of two cavities are adjusted to generate four quadrature signals for each individual cavity. Both simulation and experimental results are presented to validate the performance of this technique. The experimental results demonstrate that dynamic signals at frequencies of 100 Hz, 200 Hz, and 300 Hz with varying amplitude are successfully extracted from a dual-cavity EFPI sensor with initial lengths of 93.4803 µm and 94.0091 µm. The technique shows the potential application to measure dynamic signals in dual-cavity fiber-optic EFPI sensors.

All-sapphire fiber-optic sensor for the simultaneous measurement of ultra-high temperature and high pressure.

An all-sapphire fiber-optic extrinsic Fabry-Perot interferometric (EFPI) sensor for the simultaneous measurement of ultra-high temperature and high pressure is proposed and experimentally demonstrated. The sensor is fabricated based on all-sapphire, including a sapphire fiber, a sapphire capillary and a sapphire wafer. A femtosecond (fs) laser is employed to drill a through hole at the side wall of the sapphire capillary to allow gas flow. The sapphire fiber is inserted from one side of the sapphire capillary. The sapphire wafer is fixed at the other side of the sapphire capillary. The first Fabry-Perot (FP) cavity, composed of the end face of the sapphire fiber and the front surface of the sapphire wafer, is used for measuring pressure, while the second FP cavity, composed of the two surfaces of the sapphire wafer, is used for measuring temperature. Experimental results show that the sensor can simultaneously measure ultra-high temperature and gas pressure within the temperature range of 20 - 1400 °C and the pressure range of 0 - 5 MPa. The temperature sensitivity is 0.0033 µm/°C, and the pressure sensitivity decreases as the temperature increases, reaching 1.8016 µm/MPa and 0.3253 µm/MPa at temperatures of 20 °C and 1400 °C, respectively.

Transmission mechanism of antibiotic resistance genes and their differences between water and sediment in the Weihe River Basin.

Antibiotic resistance genes (ARGs) are emerging microbial pollutants that are regulated by many factors and pose potential threats to aquatic environments. In this study, we used network analysis, correlation analysis, and constructed models based on metagenomic sequencing results to explore the spatial patterns, impact mechanisms, transmission risks and differences in ARGs in the water and sediment of the Weihe River Basin. The findings revealed notable disparities in ARGs, mobile genetic elements (MGEs), and bacterial communities. In the sediment, the abundance of ARGs was considerably greater than that in water. Moreover, the percentage of ARGs shared by the two components reached a value of 85.8%. Through network analysis, it was determined that the presence of 16 MGEs and 20 bacterial phyla was strongly associated with ARGs (R2 > 0.7, P < 0.05). The Mantel test showed that abiotic factors including DO, pH, nutrients, and heavy metals played important roles in the distribution of ARGs (P < 0.05). A structural equation model revealed that the key factors influencing the distribution of ARGs in water were bacterial diversity and environmental parameters (standardized effects of -0.730 and -0.667), and those in sediment were bacterial diversity and MGEs (standardized effects of -0.751 and 0.851). Neutral modeling indicated that deterministic processes played an important role in the assembly of ARGs in the water of the Weihe River Basin, and stochastic processes were dominant in the sediment. There was a highly significant positive linear correlation between ARGs and pathogens, and there was more complex co-occurrence in the water than in the sediment (R2 > 0.9, P < 0.05), with stronger migration and transmission occurring. Exploring ARGs in large-scale watersheds is immensely important for elucidating their traits and transmission mechanisms and consequently paving the way for the formulation of efficient strategies to mitigate resistance threats.

[Establishment of a rapid detection method of colloidal gold immunochromatographic competition assay for organophosphorus pesticides].

Objective To establish a colloidal gold immunochromatography and develop the corresponding test strip for detecting organophosphorus compounds including omethoate, phoxim, dipterex, and parathion in fruits, vegetables and drinking water. Methods Artificial antigen molecules of organophosphorus compounds were synthesized using N-hydroxysuccinimide esters. Acetylcholinesterase antigen was prepared and purified, and the serum containing the corresponding antibody was prepared, purified, and labeled. The working parameters of the test strip were optimized, and the performance evaluation of it was conducted. Results The titer of the antisera ranged from 1:32 to 1:64, with a protein content of approximately 2 mg/mL. The purified polyclonal antibodies displayed target bands at relative molecular masses (Mr) of 25 000 and 55 000, indicating satisfactory purity. The reaction time of the test strips was between 5 to 10 minutes, with a detection limit for samples at 200 ng/mL. Both specificity and accuracy were satisfactory, and the test strip remained valid for 6 months. Conclusion A simple and rapid colloidal gold immunochromatography is established successfully for detecting several organophosphorus compounds and may be useful for on-site preliminary screening of samples in large quantities.

Mechanisms for synergistically enhancing cadmium remediation performance of biochar: Silicon activation and functional group effects.

This work proposes an advanced biochar material (ß-CD@SiBC) for controllable transformation of specific silicon (Si) forms through endogenous Si activation and functional group introduction for efficient cadmium (Cd) immobilization and removal. The maximum adsorption capacity of ß-CD@SiBC for Cd(II) reached 137.6 mg g-1 with a remarkable removal efficiency of 99 % for 200 mg L-1Cd(II). Moreover, the developed ß-CD@SiBC flow column exhibited excellent performance at the environmental Cd concentration, with the final concentration meeting the environmental standard for surface water quality (0.05 mg L-1). The remediation mechanism of ß-CD@SiBC could be mainly attributed to mineral precipitation and ion exchange, which accounted for 42 % and 29 % of the remediation effect, respectively, while functional group introduction enhanced its binding stability with Cd. Overall, this work proposes the role and principle of transformation of Si forms within biochar, providing new strategies for better utilizing endogenous components in biomass.

A study of the impact of staggered boards on corporate financialization: from the perspective of board governance.

Introduction: The objective of this study is to assess the influence of staggered boards on corporate financialization and the role that incentive and supervision mechanisms play in this process. Methods: We employ a total of 20,647 panel data samples of Chinese A-share listed companies over the period 2011-2020 to empirically test the impact of staggered boards on corporate financialization in the Chinese context. Results: The results indicate that implementing staggered boards significantly increases levels of corporate financialization. On the one hand, the implementation of a staggered board structure can exacerbate the speculative mindset and profit-driven behavior among board members, leading management to prioritize financial investments for personal gain. On the other hand, a staggered board system may also amplify managerial laziness, potentially incentivizing them to rely heavily on financial investments in order to swiftly achieve performance targets with minimal effort. Furthermore, both managerial ownership and audit supervision are found to be critical factors in mitigating this positive impact and preventing excessive financial investment behavior. Discussion: This paper offers guidance on comprehending the applicability of staggered board provisions and mitigating financial risks in enterprises.

Efficient Carrier Multiplication in Self-Powered Near-Ultraviolet γ-InSe/Graphene Heterostructure Photodetector with External Quantum Efficiency Exceeding 161.

Carrier multiplication (CM) in semiconductors, the process of absorbing a single high-energy photon to form two or more electron-hole pairs, offers great potential for the high-response detection of high-energy photons in the ultraviolet spectrum. However, compared to two-dimensional semiconductors, conventional bulk semiconductors not only face integration and flexibility bottlenecks but also exhibit inferior CM performance. To attain efficient CM for ultraviolet detection, we designed a two-terminal photodetector featuring a unilateral Schottky junction based on a two-dimensional γ-InSe/graphene heterostructure. Benefiting from a strong built-in electric field, the photogenerated high-energy electrons in γ-InSe, an ideal ultraviolet light-absorbing layer, can efficiently transfer to graphene without cooling. It results in efficient CM within the graphene, yielding an ultrahigh responsivity of 468 mA/W and a record-high external quantum efficiency of 161.2% when it is exposed to 360 nm light at zero bias. This work provides valuable insights into developing next-generation ultraviolet photodetectors with high performance and low-power consumption.

Advances on the Mechanisms and Therapeutic Strategies in Non-coding CGG Repeat Expansion Diseases.

Non-coding CGG repeat expansions within the 5' untranslated region are implicated in a range of neurological disorders, including fragile X-associated tremor/ataxia syndrome, oculopharyngeal myopathy with leukodystrophy, and oculopharyngodistal myopathy. This review outlined the general characteristics of diseases associated with non-coding CGG repeat expansions, detailing their clinical manifestations and neuroimaging patterns, which often overlap and indicate shared pathophysiological traits. We summarized the underlying molecular mechanisms of these disorders, providing new insights into the roles that DNA, RNA, and toxic proteins play. Understanding these mechanisms is crucial for the development of targeted therapeutic strategies. These strategies include a range of approaches, such as antisense oligonucleotides, RNA interference, genomic DNA editing, small molecule interventions, and other treatments aimed at correcting the dysregulated processes inherent in these disorders. A deeper understanding of the shared mechanisms among non-coding CGG repeat expansion disorders may hold the potential to catalyze the development of innovative therapies, ultimately offering relief to individuals grappling with these debilitating neurological conditions.

Genome-Wide Identification and Expression Analysis of ent-kaurene synthase-like Gene Family Associated with Abiotic Stress in Rice.

Rice (Oryza sativa) is one of the most important crops for humans. The homologs of ent-kaurene synthase (KS) in rice, which are responsible for the biosynthesis of gibberellins and various phytoalexins, are identified by their distinct biochemical functions. However, the KS-Like (KSL) family's potential functions related to hormone and abiotic stress in rice remain uncertain. Here, we identified the KSL family of 19 species by domain analysis and grouped 97 KSL family proteins into three categories. Collinearity analysis of KSLs among Poaceae indicated that the KSL gene may independently evolve and OsKSL1 and OsKSL4 likely play a significant role in the evolutionary process. Tissue expression analysis showed that two-thirds of OsKSLs were expressed in various tissues, whereas OsKSL3 and OsKSL5 were specifically expressed in the root and OsKSL4 in the leaf. Based on the fact that OsKSL2 participates in the biosynthesis of gibberellins and promoter analysis, we detected the gene expression profiles of OsKSLs under hormone treatments (GA, PAC, and ABA) and abiotic stresses (darkness and submergence). The qRT-PCR results demonstrated that OsKSL1, OsKSL3, and OsKSL4 responded to all of the treatments, meaning that these three genes can be candidate genes for abiotic stress. Our results provide new insights into the function of the KSL family in rice growth and resistance to abiotic stress.

Inhibiting HSD17B8 suppresses the cell proliferation caused by PTEN failure.

Loss of the tumor suppressor PTEN homolog daf-18 in Caenorhabditis elegans (C. elegans) triggers diapause cell division during L1 arrest. While prior studies have delved into established pathways, our investigation takes an innovative route. Through forward genetic screening in C. elegans, we pinpoint a new player, F12E12.11, regulated by daf-18, impacting cell proliferation independently of PTEN's typical phosphatase activity. F12E12.11 is an ortholog of human estradiol 17-beta-dehydrogenase 8 (HSD17B8), which converts estradiol to estrone through its NAD-dependent 17-beta-hydroxysteroid dehydrogenase activity. We found that PTEN engages in a physical interplay with HSD17B8, introducing a distinctive suppression mechanism. The reduction in estrone levels and accumulation of estradiol may arrest tumor cells in the G2/M phase of the cell cycle through MAPK/ERK. Our study illuminates an unconventional protein interplay, providing insights into how PTEN modulates tumor suppression by restraining cell division through intricate molecular interactions.

Aptamer-Based Nongenetic Reprogramming of CARs Enables Flexible Modulation of T Cell-Mediated Tumor Immunotherapy.

Innovating the design of chimeric antigen receptors (CARs) beyond conventional structures would be necessary to address the challenges of efficacy, safety, and applicability in T cell-based cancer therapy, whereas excessive genetic modification might complicate CAR design and manufacturing, and increase gene editing risks. In this work, we used aptamers as the antigen-recognition unit to develop a nongenetic CAR engineering strategy for programming the antitumor activity and specificity of CAR T cells. Our results demonstrated that aptamer-functionalized CAR (Apt-CAR) T cells could be directly activated by recognizing target antigens on cancer cells, and then impart a cytotoxic effect for cancer elimination in vitro and in vivo. The designable antigen recognition capability of Apt-CAR T cells allows for easy modulation of their efficacy and specificity. Additionally, multiple features, e.g., tunable antigen-binding avidity and the tumor microenvironment responsiveness, could be readily integrated into Apt-CAR design without T cell re-engineering, offering a new paradigm for developing adaptable immunotherapeutics.

Novel potential lncRNA biomarker in B cells indicates essential pathogenic pathway activation in patients with SLE.

OBJECTIVES: Systemic lupus erythematosus (SLE) is a highly heterogeneous disease, and B cell abnormalities play a central role in the pathogenesis of SLE. Long non-coding RNAs (lncRNAs) have also been implicated in the pathogenesis of SLE. The expression of lncRNAs is finely regulated and cell-type dependent, so we aimed to identify B cell-expressing lncRNAs as biomarkers for SLE, and to explore their ability to reflect the status of SLE critical pathway and disease activity. METHODS: Weighted gene coexpression network analysis (WGCNA) was used to cluster B cell-expressing genes of patients with SLE into different gene modules and relate them to clinical features. Based on the results of WGCNA, candidate lncRNA levels were further explored in public bulk and single-cell RNA-sequencing data. In another independent cohort, the levels of the candidate were detected by RT-qPCR and the correlation with disease activity was analysed. RESULTS: WGCNA analysis revealed one gene module significantly correlated with clinical features, which was enriched in type I interferon (IFN) pathway. Among non-coding genes in this module, lncRNA RP11-273G15.2 was differentially expressed in all five subsets of B cells from patients with SLE compared with healthy controls and other autoimmune diseases. RT-qPCR validated that RP11-273G15.2 was highly expressed in SLE B cells and positively correlated with IFN scores (r=0.7329, p<0.0001) and disease activity (r=0.4710, p=0.0005). CONCLUSION: RP11-273G15.2 could act as a diagnostic and disease activity monitoring biomarker for SLE, which might have the potential to guide clinical management.

Investigation of the formation mechanism of the pepper starch-piperine complex.

In this study, a new carrier for loading piperine was prepared using pepper starch, and its interaction mechanism was investigated. The porous pepper starch-piperine complex (PPS-PIP) showed higher loading efficiency (76.15 %) compared to the porous corn starch-piperine complex (PCS-PIP (52.34 %)). This may be ascribed to the hemispherical shell structure of porous pepper starch (PPS) compared to the porous structure of porous corn starch (PCS) based on the SEM result. PPS-PIP had smaller particle size (10.53 µm), higher relative crystallinity (38.95 %), and better thermal stability (87.45 °C) than PCS-PIP (17.37 µm, 32.17 %, 74.35 °C). Fourier transform infrared spectroscopy (FTIR) results implied that piperine not only forms a complex with amylose but may also be physically present in porous starch. This was demonstrated by the short-range order and X-ray type. Molecular dynamics simulations confirmed that hydrogen bonding is the primary interaction between amylose and piperine. Besides the formation of the amylose-piperine complex, some of the piperine is also present in physical form.

More Is Not Always Better: Impacts of AI-Generated Confidence and Explanations in Human-Automation Interaction.

OBJECTIVE: The study aimed to enhance transparency in autonomous systems by automatically generating and visualizing confidence and explanations and assessing their impacts on performance, trust, preference, and eye-tracking behaviors in human-automation interaction. BACKGROUND: System transparency is vital to maintaining appropriate levels of trust and mission success. Previous studies presented mixed results regarding the impact of displaying likelihood information and explanations, and often relied on hand-created information, limiting scalability and failing to address real-world dynamics. METHOD: We conducted a dual-task experiment involving 42 university students who operated a simulated surveillance testbed with assistance from intelligent detectors. The study used a 2 (confidence visualization: yes vs. no) × 3 (visual explanations: none, bounding boxes, bounding boxes and keypoints) mixed design. Task performance, human trust, preference for intelligent detectors, and eye-tracking behaviors were evaluated. RESULTS: Visual explanations using bounding boxes and keypoints improved detection task performance when confidence was not displayed. Meanwhile, visual explanations enhanced trust and preference for the intelligent detector, regardless of the explanation type. Confidence visualization did not influence human trust in and preference for the intelligent detector. Moreover, both visual information slowed saccade velocities. CONCLUSION: The study demonstrated that visual explanations could improve performance, trust, and preference in human-automation interaction without confidence visualization partially by changing the search strategies. However, excessive information might cause adverse effects. APPLICATION: These findings provide guidance for the design of transparent automation, emphasizing the importance of context-appropriate and user-centered explanations to foster effective human-machine collaboration.

Non-coding DNA variants for risk in lupus.

Systemic Lupus Erythematosus (SLE) is a multifactorial autoimmune disease that arises from a dynamic interplay between genetics and environmental triggers. The advent of sophisticated genomics technology has catalyzed a shift in our understanding of disease etiology, spotlighting the pivotal role of non-coding DNA variants in SLE pathogenesis. In this review, we present a comprehensive examination of the non-coding variants associated with SLE, shedding light on their role in influencing disease risk and progression. We discuss the latest methodological advancements that have been instrumental in the identification and functional characterization of these genomic elements, with a special focus on the transformative power of CRISPR-based gene-editing technologies. Additionally, the review probes into the therapeutic opportunities that arise from modulating non-coding regions associated with SLE. Through an exploration of the complex network of non-coding DNA, this review aspires to decode the genetic puzzle of SLE and set the stage for groundbreaking gene-based therapeutic interventions and the advancement of precision medicine strategies tailored to SLE management.

Peer Stress Spills Over to Family Stress in the Context of Emotion Regulation Difficulties: A Daily Diary Study with Chinese Adolescents.

Conflict in peer and family relationships becomes more common in the adolescent period when compared to previous developmental periods. These typical developmental challenges can be exacerbated in the context of poor emotion regulation skills. Using daily diary data, the current study examined the stress spillover effects of peer and family stress on one another, as well as the moderating role of emotion regulation challenges (i.e., emotional inhibition, dysregulation). A sample of 310 Chinese adolescents (Mage = 13.02 years, SD = 0.76 years, 50.7% boys) completed an initial measure of emotion regulation difficulties, then reported on peer and family stress for 10 consecutive weekdays. Results indicated that there was an overall same-day peer stress spillover effect in which adolescents' peer stress on a given day was negatively associated with later conflictual interactions with their parents. Further, the relation between peer stress and same- and next-day family stress was exacerbated in the context of high levels of emotional inhibition. Family stress did not significantly relate to next-day peer stress, nor was this association moderated by difficulties with emotion regulation. These results highlight the temporal sequence of daily peer-to-family stress spillover. Though emotional inhibition may be culturally adaptive for maintaining interpersonal harmony, it can be maladaptive in managing stress for Chinese adolescents.