Single-Cell RNA-Sequencing of Soybean Reveals Transcriptional Changes and Antiviral Functions of GmGSTU23 and GmGSTU24 in Response to Soybean Mosaic Virus.

Soybean mosaic virus (SMV) stands as a prominent and widespread threat to soybean (Glycine max L. Merr.), the foremost legume crop globally. Attaining a thorough comprehension of the alterations in the transcriptional network of soybeans in response to SMV infection is imperative for a profound insight into the mechanisms of viral pathogenicity and host resistance. In this investigation, we isolated 50 294 protoplasts from the newly developed leaves of soybean plants subjected to both SMV infection and mock inoculation. Subsequently, we utilized single-cell RNA sequencing (scRNA-seq) to construct the transcriptional landscape at a single-cell resolution. Nineteen distinct cell clusters were identified based on the transcriptomic profiles of scRNA-seq. The annotation of three cell types-epidermal cells, mesophyll cells, and vascular cells-was established based on the expression of orthologs to reported marker genes in Arabidopsis thaliana. The differentially expressed genes between the SMV- and mock-inoculated samples were analyzed for different cell types. Our investigation delved deeper into the tau class of glutathione S-transferases (GSTUs), known for their significant contributions to plant responses against abiotic and biotic stress. A total of 57 GSTU genes were identified by a thorough genome-wide investigation in the soybean genome G. max Wm82.a4.v1. Two specific candidates, GmGSTU23 and GmGSTU24, exhibited distinct upregulation in all three cell types in response to SMV infection, prompting their selection for further research. The transient overexpression of GmGSTU23 or GmGSTU24 in Nicotiana benthamiana resulted in the inhibition of SMV infection, indicating the antiviral function of soybean GSTU proteins.

Research Progress on the Preparation and Function of Antioxidant Peptides from Walnuts.

Food-derived peptides have good antioxidant activity and are highly safe for humans; consequently, there has been continuous growth in research on antioxidants, with potential applications in food, medicine, cosmetics, and other fields. Among food-derived peptides, walnut-derived peptides have attracted increasing attention as food-derived peptides rich in eight essential amino acids. This review summarizes the progress made in the development and identification of antioxidant peptides in walnut proteins. This article mainly describes the interaction between reactive oxygen species and cellular antioxidant products, modulation of enzyme content and activity, and regulation of the redox signaling pathways and analyzes the mechanisms of reduction in oxidative stress. Finally, the complex structure-activity relationships of walnut-derived peptides are analyzed based on their amino acid composition and secondary structure of the polypeptides. This review provides a theoretical basis for the production of walnut-derived antioxidant peptides and could help promote the development of the walnut industry.

GsRSS3L, a Candidate Gene Underlying Soybean Resistance to Seedcoat Mottling Derived from Wild Soybean (Glycine soja Sieb. and Zucc).

Soybeans are a major crop that produce the best vegetable oil and protein for use in food and beverage products worldwide. However, one of the most well-known viral infections affecting soybeans is the Soybean Mosaic Virus (SMV), a member of the Potyviridae family. A crucial method for preventing SMV damage is the breeding of resistant soybean cultivars. Adult resistance and resistance of seedcoat mottling are two types of resistance to SMV. Most studies have focused on adult-plant resistance but not on the resistance to seedcoat mottling. In this study, chromosome segment-substituted lines derived from a cross between Suinong14 (cultivated soybean) and ZYD00006 (wild soybean) were used to identify the chromosome region and candidate genes underlying soybean resistance to seed coat mottling. Herein, two quantitative trait loci (QTLs) were found on chromosome 17, and eighteen genes were found in the QTL region. RNA-seq was used to evaluate the differentially expressed genes (DEGs) among the eighteen genes located in the QTLs. According to the obtained data, variations were observed in the expression of five genes following SMV infection. Furthermore, Nicotiana benthamiana was subjected to an Agrobacterium-mediated transient expression assay to investigate the role of the five candidate genes in SMV resistance. It has also been revealed that Glyma.17g238900 encoding a RICE SALT SENSITIVE 3-like protein (RSS3L) can inhibit the multiplication of SMV in N.benthamiana. Moreover, two nonsynonymous single-nucleotide polymorphisms (SNPs) were found in the coding sequence of Glyma.17g238900 derived from the wild soybean ZYD00006 (GsRSS3L), and the two amino acid mutants may be associated with SMV resistance. Hence, it has been suggested that GsRSS3L confers seedcoat mottling resistance, shedding light on the mechanism of soybean resistance to SMV.

Physicochemical and Colon Cancer Cell Inhibitory Properties of Theabrownins Prepared by Weak Alkali Oxidation of Tea Polyphenols.

Existing studies on the biological activity of theabrownins are not based on their free state but on the complexes of theabrownins, polysaccharides, proteins, and flavonoids. In this study, theabrownins (TBs-C) were prepared by weak alkali oxidation of tea polyphenols. The ultraviolet-visible scanning spectrum of TBs-C showed two characteristic absorption peaks at 203 and 270 nm. The zeta potential of the TBs-C aqueous solution was negative, and the values varied from - 6.26 to -19.55 mV with a solution pH of 3-9. Storage conditions of pH 5.0-7.0 and around 25 °C were beneficial for the physical and chemical stability of the TBS-C solution. Cells were treated with series concentrations and examined by MTT, HE staining, PI immunofluorescence staining, flow cytometry, and real-time PCR to investigate the antiproliferative effect of TBs-C on human colon cancer HT-29 cells. The results showed that TBs-C, particularly at 500 µg/mL, inhibited cell growth. TBs-C induced HT-29 cell apoptosis, as confirmed by morphological changes, nucleus propidium iodide staining, and distributions of the cell cycle. The apoptotic mechanism may be due to the intracellular redox imbalance induced by TBs-C.

Hepatotoxicity and lipid metabolism disorders of 8:2 polyfluoroalkyl phosphate diester in zebrafish: In vivo and in silico evidence.

8:2 polyfluoroalkyl phosphate diester (8:2 diPAP) has been shown to accumulate in the liver, but whether it induces hepatotoxicity and lipid metabolism disorders remains largely unknown. In this study, zebrafish embryos were exposed to 8:2 diPAP for 7 d. Hepatocellular hypertrophy and karyolysis were noted after exposure to 0.5 ng/L 8:2 diPAP, suggesting suppressed liver development. Compared to the water control, 8:2 diPAP led to significantly higher triglyceride and total cholesterol levels, but markedly lower levels of low-density lipoprotein, implying disturbed lipid homeostasis. The levels of two peroxisome proliferator activated receptor (PPAR) subtypes (pparα and pparγ) involved in hepatotoxicity and lipid metabolism were significantly upregulated by 8:2 diPAP, consistent with their overexpression as determined by immunohistochemistry. In silico results showed that 8:2 diPAP formed hydrogen bonds with PPARα and PPARγ. Among seven machine learning models, Adaptive Boosting performed the best in predicting the binding affinities of PPARα and PPARγ on the test set. The predicted binding affinity of 8:2 diPAP to PPARα (7.12) was higher than that to PPARγ (6.97) by Adaptive Boosting, which matched well with the experimental results. Our results revealed PPAR - mediated adverse effects of 8:2 diPAP on the liver and lipid metabolism of zebrafish larvae.

Effects of a Parent-Child Single-Session Growth Mindset Intervention on Adolescent Depression and Anxiety Symptoms: Protocol of a 3-Arm Waitlist Randomized Controlled Trial.

BACKGROUND: Depression and anxiety are common mental health problems among adolescents worldwide. Extant research has found that intelligence, emotion, and failure-is-debilitating beliefs (fixed mindsets) are closely related to more depression and anxiety symptoms, hopelessness, and suicidality. Recent research also points to the importance of parental mindset, which can strongly influence children's affect, behavior, and mental health. However, the effects of parent-child mindset interventions on a child's internalizing problems have not yet been empirically examined. As recent evidence has shown the promise of single-session interventions in reducing and preventing youth internalizing problems, this study develops and examines a parent and child single-session intervention on mindsets of intelligence, failure, and emotion (PC-SMILE) to tackle depression and anxiety in young people. OBJECTIVE: Using a 3-arm randomized controlled trial, this study will examine the effectiveness of PC-SMILE in reducing depression and anxiety symptoms among children. We hypothesize that compared to the waitlist control group, the PC-SMILE group and child single-session intervention on mindsets of intelligence, failure, and emotion (C-SMILE) group will significantly improve child depression and anxiety (primary outcome) and significantly improve secondary outcomes, including children's academic self-efficacy, hopelessness, psychological well-being, and parent-child interactions and relationships, and the PC-SMILE is more effective than the C-SMILE. METHODS: A total of 549 parent-child dyads will be recruited from 8 secondary schools and randomly assigned to either the PC-SMILE intervention group, the C-SMILE intervention group, or the no-intervention waitlist control group. The 45-minute interventions include parent-version and child-version. Both parents and students in the PC-SMILE group receive the intervention. Students in C-SMILE group receive intervention and their parents will receive intervention after all follow-up ends. Students in 3 groups will be assessed at 3 time points, baseline before intervention, 2 weeks post intervention, and 3 months post intervention, and parents will be assessed in baseline and 3-month follow-up. The intention-to-treat principle and linear-regression-based maximum likelihood multilevel models will be used for data analysis. RESULTS: Recruitment started in September 2023. The first cohort of data collection is expected to begin in May 2024 and the second cohort will begin in September 2024. The final wave of data is expected to be collected by the end of the first quarter of 2025. The results are expected to demonstrate improved anxiety and depression among students assigned to the intervention condition, as well as the secondary outcomes compared to those in the control group. The efficacy and effectiveness of the intervention will be discussed. CONCLUSIONS: This study is the first attempt to develop a web-based single-session intervention for students and their parents to enhance their well-being in Hong Kong and beyond, which potentially contributes to providing evidence-based recommendations for the implementation of brief digital parent-child interventions. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): PRR1-10.2196/63220.

A chromosome-scale assembly of the early-flowering Prunus campanulata and comparative genomics of cherries.

Prunus campanulata is an important flowering cherry germplasm of high ornamental value. Given its early-flowering phenotypes, P. campanulata could be used for molecular breeding of ornamental species and fruit crops belonging to the subgenus Cerasus. Here, we report a chromosome-scale assembly of P. campanulata with a genome size of 282.6 Mb and a contig N50 length of 12.04 Mb. The genome contained 24,861 protein-coding genes, of which 24,749 genes (99.5%) were functionally annotated, and 148.20 Mb (52.4%) of the assembled sequences are repetitive sequences. A combination of genomic and population genomic analyses revealed a number of genes under positive selection or accelerated molecular evolution in P. campanulata. Our study provides a reliable genome resource, and lays a solid foundation for genetic improvement of flowering cherry germplasm.

Probing the bioconcentration and metabolism disruption of bisphenol A and its analogues in adult female zebrafish from integrated AutoQSAR and metabolomics studies.

Plenty of emerging bisphenol A (BPA) substitutes rise to wait for assessment of bioconcentration and metabolism disruption. Computational methods are useful to fill the data gap in chemical risk assessment, such as automated quantitative structure-activity relationship (AutoQSAR). It is not clear how AutoQSAR performs in predicting the bioconcentration factor (BCF) in adult zebrafish. Herein, AutoQSAR was used to predict the logBCFs of BPA, bisphenol AF (BPAF), bisphenol B, bisphenol F and bisphenol S (BPS). For the test set, a linear relationship was shown between the observed and predicted logBCFs with a slope of 0.97. The predicted logBCFs of these five bisphenols were quite close to their experimental data with a slope of 0.94, suggesting better performance than directed message passing neural networks and EPI Suite with a slope of 0.69 and 0.61, respectively. Thus, AutoQSAR is powerful in modeling logBCFs in fish with minimal time and expertise. To link bioconcentration with metabolic effects, female zebrafish were exposed to BPA, BPAF and BPS for metabolomics analysis. BPA caused a significant disturbance in amino acid metabolism, while BPAF and BPS significantly altered another three metabolic pathways, showing chemical-specific responses. BPAF with the highest logBCF elicited the strongest metabolomic responses reflected by the metabolic effect level index, followed by BPA and BPS. Thus, BPAF and BPS elicited higher or similar metabolism disruption compared with BPA in female zebrafish, respectively, reflecting consequences of bioconcentration.

Effects of hydrocolloids as fat-replacers on the physicochemical and structural properties of salt-soluble protein isolated from water-boiled pork meatballs.

Konjac glucomannan (KGM), xanthan gum (XG), guar gum (GG), and κ-carrageenan (KC), as substituent, are commonly used in ground pork products. Here, the content of these (0.5, 1.0, 1.5, and 2.0%, w/w) on the gel properties, thermal properties, and interaction forces of salt-soluble protein (SSP) isolated from water-boiled pork meatballs were investigated. We found 1.0% KGM, 0.5% XG, 0.5-2.0% GG, and 0.5-2.0% KC to water-boiled pork meatballs exerted a positive effect on the denaturation temperature, hydrogen bonds, hydrophobic interactions, disulfide bonds, α-helix, and ß-sheet content of SSP, as well as the strength and storage modulus of the modified protein gel. The addition of these hydrocolloids with the addition of 1.0% aggregated myosin and actin, led to the enhancement of the bands corresponding to myosin heavy chain and actin. The prediction model of gel strength showed that the gel strength was negatively correlated with Tpeak1, Tpeak2, and ionic bond. This study provides theoretical guidance for improving the application of hydrocolloids in pork-based foods.

Single-session intervention on growth mindset on negative emotions for university student mental health (U-SIGMA): a protocol of two-armed randomized controlled trial.

BACKGROUND: The university years are a developmentally crucial phase and a peak period for the onset of mental disorders. The beliefs about the changeability of negative emotion may play an important role in help-seeking. The brief digital growth mindset intervention is potentially scalable and acceptable to enhance adaptive coping and help-seeking for mental health needs in university students. We adapted the Single-session Intervention on Growth Mindset for adolescents (SIGMA) to be applied in university students (U-SIGMA). This protocol introduces a two-armed waitlist randomized controlled trial study to examine the effectiveness and acceptability of U-SIGMA in promoting help-seeking among university students in the Greater Bay Area. METHODS: University students (N = 250, ages 18-25) from universities in the Greater Bay Area will be randomized to either the brief digital growth mindset intervention group or the waitlist control group. Participants will report on the mindsets of negative emotions, perceived control over anxiety, attitude toward help-seeking, physical activity, hopelessness, psychological well-being, depression, anxiety, and perceived stress at baseline and the 2-week and 8-week follow-ups through web-based surveys. A 30-min digital intervention will be implemented in the intervention group, with a pre- and post-intervention survey collecting intervention feedback, while the control group will receive the link for intervention after 8 weeks. DISCUSSION: This protocol introduces the implementation plan of U-SIMGA in multi-cities of the Greater Bay Area. The findings are expected to help provide pioneer evidence for the effectiveness and acceptability of the brief digital intervention for university students in the Chinese context and beyond and contribute to the development of accessible and effective prevention and early intervention for university students' mental health. TRIAL REGISTRATION: HKU Clinical Trials Registry: HKUCTR-3012; Registered 14 April 2023.  http://www.hkuctr.com/Study/Show/7a3ffbc0e03f4d1eac0525450fc5187e .

The Mental Health Impact of the COVID-19 Pandemic on Older Adults in China: A Systematic Review.

Considered at a high risk during the COVID-19 pandemic, older adults in China not only face the disadvantages caused by their relatively low immune systems, but also the challenges brought about by the complex psychological environment in which they spend this special period of their life. However, a thorough study on the impact of the pandemic on older adults' mental health in China remains scant. Hence, this research aimed to investigate the question: What are the mental health outcomes and associated risk factors of the COVID-19 pandemic on older adults in China? Two Chinese academic databases (China National Knowledge Infrastructure and WANFANG DATA) as well as six English academic databases (PubMed, Scopus, Web of Science, MEDLINE, Social Science, and Google Scholar) were searched while following PRISMA guidelines. Studies were selected according to the predetermined inclusion criteria. Further, relatively high detective rates of mental health disorders, including anxiety symptoms (4.9% to 48.6%), depression symptoms (13.8% to 58.7%), hypochondria (11.9%), suicidal ideation (4.1%), along with worries and fear (55.7%) were all reported. The COVID-19 pandemic has presented a threat to not only the physical, but also the psychological health of Chinese older adults. The most common risk factors of psychological distress among Chinese older adults were found in female gender, living in rural areas, coexisting chronic diseases, and insufficient knowledge about the COVID pandemic. As a result, government policy and psychological guidelines that are created in order to alleviate the adverse effects of COVID-19 on older adults' mental health, need to be further developed.

Genomic population structure and local adaptation of the wild strawberry Fragaria nilgerrensis.

The crop wild relative, Fragaria nilgerrensis, is adapted to a variety of diverse habitats across its native range in China. Thus, discoveries made in this species could serve useful to guide the development of new superior strawberry cultivars that are resilient to new or variable environments. However, the genetic diversity and genetic architecture of traits in this species underlying important adaptive traits remain poorly understood. Here, we used whole-genome resequencing data from 193 F. nilgerrensis individuals spanning the distribution range in China to investigate the genetic diversity, population structure and the genomic basis of local adaptation. We identified four genetic groups, with the western group located in Hengduan Mountains exhibited the highest genetic diversity. Redundancy analysis suggests that both environment and geographic variables shaped a significant proportion of genomic variation. Our analyses revealed that the environmental difference explains more of the observed genetic variation than geographic distance. This suggests that adaptation to distinct habitats, unique combination of abiotic factors, likely drove genetic differentiation. Lastly, by implementing selective sweeps scans and genome-environment association analysis throughout the genome, we identified the genetic variation associated with local adaptation and investigated the functions of putative candidate genes in F. nilgerrensis.

Characterization of Theabrownins Prepared From Tea Polyphenols by Enzymatic and Chemical Oxidation and Their Inhibitory Effect on Colon Cancer Cells.

Theabrownins (TBs) are prepared from dark tea and contain a large number of complex heterogeneous components, such as carbohydrates, proteins, and flavonoids, which are difficult to remove. In addition, some toxic and harmful extraction solvents are used to purify TBs. These obstacles hinder the utilization and industrialization of TBs. In this study, tea polyphenols were used as substrates and polyphenol oxidase and sodium bicarbonate (NaHCO3) were used successively to prepare theabrownins (TBs-E). The UV-visible characteristic absorption peaks of the TBs-E were located at 203 and 270 nm and Fourier-transform IR analysis showed that they were polymerized phenolic substances containing the hydroxy and carboxyl groups. The TBs-E aqueous solution was negatively charged and the absolute values of the zeta potential increased with increasing pH. A storage experiment showed that TBs-E were more stable at pH 7.0 and in low-temperature environments around 25°C. HT-29 human colon cancer cells were used to evaluate the biological activity of TBs-E through 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide (MTT), H&E staining, propidium iodide immunofluorescent staining, flow cytometry, and real-time PCR assays. The TBs-E significantly inhibited cell growth and caused late apoptosis, particularly at the dose of 500 µg/ml. The TBs-E markedly reduced the expression of antioxidant enzyme genes and increased the generation of reactive oxygen species to break the redox balance, which may have led to cell damage and death. These results will promote research and industrialization of TBs.

Lignification and lignin heterogeneity for various age classes of bamboo (Phyllostachys pubescens) stems.

The lignification process and lignin heterogeneity of fibre, vessel and parenchyma cell walls for various age classes of bamboo stems of Phyllostachys pubescens Mazel were investigated. It was shown that protoxylem vessels lignified in the early stage of vascular bundle differentiation, metaxylem vessel and fibre walls initiated lignification from the middle lamella and cell corners after the completion of vascular bundle differentiation. Most of the parenchyma cell walls lignified after the stem reached its full height, while a few parenchyma cells remained non-lignified even in the mature culm. The cell walls of fibres and most parenchyma cells thickened further during the stem growth to form polylamellate structure and the lignification process of these cells may last even up to 7 years. The fibre walls were rich in guaiacyl lignin in the early stage of lignification, and lignin rich in syringyl units were deposited in the later stage. Vessel walls mainly contained guaiacyl lignin, while both guaiacyl and syringyl lignin were present in the fibre and parenchyma cell walls.

Casparian strips in needles of Pinus bungeana: isolation and chemical characterization.

By using cell wall degrading enzymes, Casparian strips were for the first time isolated from Pinus bungeana needle endodermis. They appeared as a fine network, similar to those isolated from roots. Fourier transform infrared spectroscopic analysis provided evidence that the Casparian strips were impregnated with lignin, suberin, cellulose and cell wall proteins.

Inhibition of RNA and protein synthesis in pollen tube development of Pinus bungeana by actinomycin D and cycloheximide.

* The effects of actinomycin D and cycloheximide on RNA and protein synthesis were investigated during pollen tube development of Pinus bungeana. * RNA and protein contents, protein expression patterns, cell wall components and ultrastructural changes of pollen tubes were studied using spectrophotometry, SDS-PAGE electrophoresis, Fourier transformed infrared (FTIR) microspectroscopy and transmission electron microscopy (TEM). * Pollen grains germinated in the presence of actinomycin D, but tube elongation and RNA synthesis were inhibited. By contrast, cycloheximide inhibited pollen germination and protein synthesis, induced abnormal tube morphology, and retarded the tube growth rate. SDS-PAGE analysis showed that protein expression patterns changed distinctly, with some proteins being specific for each phase. FTIR microspectroscopy established significant changes in the chemical composition of pollen tube walls. TEM analysis revealed the inhibitors caused disintegration of organelles involved in the secretory system. * These results suggested RNA necessary for pollen germination and early tube growth were present already in the pollen grains before germination, while the initiation of germination and the maintenance of pollen tube elongation depended on continuous protein synthesis.