Integrated agronomic, physiological, microstructure, and whole-transcriptome analyses reveal the role of biomass accumulation and quality formation during Se biofortification in alfalfa.

Se-biofortified agricultural products receive considerable interest due to the worldwide severity of selenium (Se) deficiency. Alfalfa (Medicago sativa L.), the king of forage, has a large biomass, a high protein content, and a high level of adaptability, making it a good resource for Se biofortification. Analyses of agronomic, quality, physiological, and microstructure results indicated the mechanism of biomass increase and quality development in alfalfa during Se treatment. Se treatment effectively increased Se content, biomass accumulation, and protein levels in alfalfa. The enhancement of antioxidant capacity contributes to the maintenance of low levels of reactive oxygen species (ROS), which, in turn, serves to increase alfalfa's stress resistance and the stability of its intracellular environment. An increase in the rate of photosynthesis contributes to the accumulation of biomass in alfalfa. To conduct a more comprehensive investigation of the regulatory networks induced by Se treatment, the transcriptome sequencing of non-coding RNA (ncRNA) was employed to compare 100 mg/kg Se treatment and control groups. The analysis identified 1,414, 62, and 5 genes as DE-long non-coding RNAs (DE-lncRNA), DE-microRNAs (DE-miRNA), and DE-circular RNA (DE-circRNA), respectively. The function of miRNA-related regulatory networks during Se biofortification in alfalfa was investigated. Subsequent enrichment analysis revealed significant involvement of transcription factors, DNA replication and repair mechanisms, photosynthesis, carbohydrate metabolism, and protein processing. The antioxidant capacity and protein accumulation of alfalfa were regulated by the modulation of signal transduction, the glyoxalase pathway, proteostasis, and circRNA/lncRNA-related regulatory networks. The findings offer new perspectives on the regulatory mechanisms of Se in plant growth, biomass accumulation, and stress responses, and propose potential strategies for enhancing its utilization in the agricultural sector.

Triple-kernel gated attention-based multiple instance learning with contrastive learning for medical image analysis.

In machine learning, multiple instance learning is a method evolved from supervised learning algorithms, which defines a "bag" as a collection of multiple examples with a wide range of applications. In this paper, we propose a novel deep multiple instance learning model for medical image analysis, called triple-kernel gated attention-based multiple instance learning with contrastive learning. It can be used to overcome the limitations of the existing multiple instance learning approaches to medical image analysis. Our model consists of four steps. i) Extracting the representations by a simple convolutional neural network using contrastive learning for training. ii) Using three different kernel functions to obtain the importance of each instance from the entire image and forming an attention map. iii) Based on the attention map, aggregating the entire image together by attention-based MIL pooling. iv) Feeding the results into the classifier for prediction. The results on different datasets demonstrate that the proposed model outperforms state-of-the-art methods on binary and weakly supervised classification tasks. It can provide more efficient classification results for various disease models and additional explanatory information.

Factors affecting physical examinations among patients with severe mental disorders in Shaoxing City / 预防医学

Objective@#To investigate the proportion of physical examinations among patients with severe mental disorders and its influencing factors in Shaoxing City, Zhejiang Province, so as to provide the evidence for improving the proportion of physical examinations among patients with severe mental disorders. @*Methods @#The epidemiological and clinical features of patients with severe mental disorders included in community management in Shaoxing City in 2022 were collected from Zhejiang Provincial Severe Mental Disorder Management Information System, including demographics, disease diagnosis and treatment, physical examination, and rescue and assistance. Factors affecting the physical examination were identified among patients with severe mental disorders using a multivariable logistic regression model. @*Results@#A total of 25 468 patients with severe mental disorders were enrolled in Shaoxing City in 2022, including 12 151 males and 13 317 females, with a male to female ratio of 0.91∶1, and the participants had a mean age of (54.34±14.71) years. Schizophrenia was the predominant type of severe mental disorders (15 419 cases, 60.54%), and 21 374 subjects participating in the physical examinations in 2022 (83.92%). Multivariable logistic regression analysis showed that female (OR=0.901, 95%CI: 0.832-0.975), urban areas (OR=0.506, 95%CI: 0.468-0.547), mental disorders due to epilepsy (OR=1.779, 95%CI: 1.104-2.866), hospitalized treatment (6 to 10 times, OR=0.523, 95%CI: 0.401-0.681; 11 times and more, OR=0.177, 95%CI: 0.108-0.288), special diseases in outpatient (OR=1.738, 95%CI: 1.597-1.891), receiving medical assistance (OR=2.851, 95%CI: 2.616-3.107), targets of the community care and assistance groups (OR=1.653, 95%CI: 1.471-1.857) and guardian (spouse, OR=1.777, 95%CI: 1.513-2.086; children, OR=1.277, 95%CI: 1.069-1.526; parents, OR=1.342, 95%CI: 1.143-1.576) were statistically associated with the proportion of physical examinations. @*Conclusions@#The proportion of health examinations was 83.92% among patients with severe mental disorders in Shaoxing City in 2022. Gender, residence, guardian, disease diagnosis, times of hospitalized treatment, medical assistance, special diseases in outpatients and target of community care and assistance groups were factors affecting health examinations among patients with severe mental disorders.

Integrated eco-physiological, biochemical, and molecular biological analyses of selenium fortification mechanism in alfalfa.

MAIN CONCLUSION: Foliar Se (IV) application at 100 mg/kg can act as a positive bio-stimulator of redox, photosynthesis, and nutrient metabolism in alfalfa via phenotypes, nutritional compositions, biochemistry, combined with transcriptome analysis. Selenium (Se) is an essential element for mammals, and plants are the primary source of dietary Se. However, Se usually has dual (beneficial/toxic) effects on the plant itself. Alfalfa (Medicago sativa L.) is one of the most important forage resources in the world due to its high nutritive value. In this study, we have investigated the effects of sodium selenite (Se (IV)) (0, 100, 200, 300, and 500 mg/kg) on eco-physiological, biochemical, and transcriptional mechanisms in alfalfa. The phenotypic and nutritional composition alterations revealed that lower Se (IV) (100 mg/kg) levels positively affected alfalfa; it enhanced the antioxidant activity, which may contribute to redox homeostasis and chloroplast function. At 100 mg/kg Se (IV) concentration, the H2O2, and malondialdehyde (MDA) contents decreased by 36.72% and 22.62%, respectively, whereas the activity of glutathione peroxidase (GPX) increased by 31.10%. Se supplementation at 100 mg/kg increased the plant pigments contents, the light-harvesting capacity of PSII (Fv/Fm) and PSI (ΔP700max), and the carbon fixation efficiency, which was demonstrated by enhanced photosynthesis (37.6%). Furthermore, alfalfa shifted carbon flux to protein synthesis to improve quality at 100 mg/kg of Se (IV) by upregulating carbohydrate and amino acid metabolic genes. On the contrary, at 500 mg/kg, Se (IV) became toxic. Higher Se (IV) disordered the plant antioxidant system, increasing H2O2 and MDA by 14.2 and 4.3%, respectively. Moreover, photosynthesis was inhibited by 20.2%, and more structural substances, such as lignin, were synthesized. These results strongly suggest that Se (IV) at a concentration of 100 mg/kg act as the positive bio-stimulator of redox metabolism, photosynthesis, and nutrient in alfalfa.

Transcriptome analysis revealed accumulation-assimilation of selenium and physio-biochemical changes in alfalfa (Medicago sativa L.) leaves.

BACKGROUND: Selenium (Se) is an increasing concern for investigators predominantly because of its consumption in the human body mainly from crops. As the fourth largest plant crop globally, alfalfa is one of the most important forages. Alfalfa was fertilized with selenium(IV) (Se(IV)) under field conditions to study the accumulation and assimilation of Se(IV) and to assess the impact of Se fertilization. RESULTS: It was analyzed that the physio-biochemistry, Se species, combined with transcriptome after spraying Se(IV) at different times (0, 12, and 48 h). 9402 and 12 607 differentially expressed genes (DEGs) were identified at 12 h (versus 0 h) and 48 h (versus 12 h). DEG functional enrichments proposed two time-specific biological processes: Se(IV) accumulation was the primary process at 0-12 h, and its assimilation mainly occurred during 12-48 h. This was further proved by the separation of various Se speciation at different times. It showed that Se-supplementation also affected the soluble protein, soluble sugar, pigment contents and antioxidant capacity. Selenium-biofortification could improve the stress resistance of alfalfa by enhancing antioxidant system to scavenge reactive oxygen species (e.g. hydrogen peroxide) and boosting carbohydrate metabolism. CONCLUSION: By integrating physio-biochemistry, Se-related metabolites, and transcriptome under Se(IV) treatment, this study provides data to guide further work on Se-fortification in alfalfa. © 2022 Society of Chemical Industry.

Relative contribution of environmental medium and internal organs to lead accumulation of wheat grain.

Lead (Pb) pollution in wheat has received considerable research attention globally due to its persistence and ease of accumulation, posing severe health risks to humans. This study explored the relative contribution of the environmental medium (atmospheric deposition and soil) and wheat internal organs to Pb accumulation in wheat grains, using field experiments by contrasting treatments. The concentration and bioavailability of Pb in the soil were significantly lower than those of atmospherically deposited Pb (P < 0.05). Pb accumulation rate in wheat grains was consistent with the grain filling rate, which first increased and then decreased, reaching the highest level at the middle filling stage. Pb isotope analysis showed that atmospheric deposition was the main source of Pb in the shoots of wheat plants, contributing more than 80.0% of Pb in grains. Although the roots had the highest Pb concentration, the spikes had the greatest relative contribution (58.4%) to Pb accumulation in the wheat grains, followed by that of the leaves (24.5%), whereas the contribution of roots was the lowest (17.1%) among all plant organs. In addition, among all leaves, the contribution of flag leaves to Pb accumulation in the grain was higher than the cumulative contribution of all other leaves, where flag leaves and other leaves contributed 13.8% and 10.7%, respectively. Collectively, the absorption of atmospherically deposited Pb by wheat spikes is the leading cause of Pb pollution in wheat grains. These results may aid in formulating strategies to reduce Pb concentration in grains and ensure food quality and safety.

Comparative proteomics analysis of the responses to selenium in selenium-enriched alfalfa (Medicago sativa L.) leaves.

The mass of leaves and the chlorophyll and selenium content of alfalfa can be increased by the foliar spraying of selenite. To better understand the relationship between changes in the expression of specific proteins and the various metabolic and regulatory pathways affected by selenium treatment, labeling with Tandem Mass Tags (TMT) was used as a proteomics technique to compare control leaves with those enriched with Se. A total of 8,411 proteins were identified, the expression levels of 195 of which were significantly modified, 67 significantly up-regulated and 128 significantly down-regulated. Using gene functional classification and metabolic pathway annotation, selenium treatment was found to have a significant impact on metabolic processes. The energy and substances produced by the metabolic processes of a variety of carbohydrates, lipids, and amino acids, and the metabolism of carbon may be responsible for increasing the yield of alfalfa leaves. Administration of selenium substantially influenced Se-responsive proteins, including ABC transporter G family member 36, Probable glutathione S-transferase and cysteine tRNA ligase. Selenium treatment may also enhance photosynthesis and the defense response of cells. Furthermore, protein ubiquitination also played an important role in the selenium response of alfalfa leaves. In summary, a basic analysis of the selenium response pathway in alfalfa leaves at the proteomics level was conducted, which may assist in a more detailed elucidation of selenium enrichment in alfalfa in the future.

A methodology study on chemical and molecular structure imaging in modified forage leaf tissue with cutting-edge synchrotron-powered technology (SR-IMS) as a potential research tool.

To date there is no any study on imaging molecular chemistry and chemical structure of biotech-modified plant tissue on a molecular basis. The objective of this methodology study was to apply a non-invasive and non-destructive synchrotron powered technology - SR-IMS to image molecular chemistry of the modified forage leaf tissue. The infrared molecular vibrational microspectroscopy powered with synchrotron light at Advanced Light Source (ALS, Lawrence Berkeley National Lab, Berkeley, California, Dept. of Energy, USA) were applied. The synchrotron beamline time was arranged by National Synchrotron Light Source (Scientist Dr. Lisa Miller, Brookhaven National Lab, Dept. of Energy, USA). The various molecular functional groups in the forage tissue included CH symmetric and asymmetric regions, amides I and II regions, structure and non-structure CHO regions, carbonyl ester region with peak areas at ca. 3644-3000 cm-1, ca 3005-2979 cm-1, ca. 1722-1483 cm-1, ca. 1488-1412 cm-1, ca. 1296-1189 cm-1, and ca. 1194-951 cm-1. The spectral peak area ratio imaging of chemical functional groups were also studied which included the ratio of peak area under ca. 1722-1483 cm-1 to peak area under ca. 3005-2979 cm-1 and the ratio of peak area under ca. 1722-1483 cm-1 to peak area under ca. 1194-951 cm-1. The results showed that the advanced synchrotron-based technology - SR-IMS was able to image the forage tissue at an ultra-highly resolution within intact tissue within cellular and subcellular dimension. It revealed the forage tissue in a molecular chemical sense and provided an insight on nutrient properties and their molecular structure as well as chemical features. In conclusion, the synchrotron-radiation SR-IMS is able to image molecular structure of the forage leaf tissue at an ultra-highly resolution. The advanced SR-IMS technique could provide leaf tissue four kinds of information simultaneously: tissue structure, tissue chemistry, tissue nutrients, and tissue environment of forage.

Dissolution of kaolinite induced by citric, oxalic, and malic acids.

Kaolinite is a dominant clay mineral in the soils in tropical and subtropical regions, and its dissolution has an influence on a variety of soil properties. In this work, kaolinite dissolution induced by three kinds of low-molecular-weight organic acid, i.e., citric, oxalic, and malic acids, was evaluated under far-from-equilibrium conditions. The rates of kaolinite dissolution depended on the kind and concentration of organic acids, with the sequence R(oxalate)>R(citrate)>R(malate). Chemical calculation showed the change in concentration of organic ligand relative to change in concentration of organic acid in suspensions of kaolinite and organic acid. The effect of organic acid on kaolinite dissolution was modeled by species of organic anionic ligand. For oxalic acid, L(2-)(oxalic) and HL(-)(oxalic) jointly enhanced the dissolution of kaolinite, but for malic and citric acids, HL(-)(malic) and H2L-(citric) made a higher contribution to the total dissolution rate of kaolinite than L(2-)(malic) and L(3-)(citric), respectively. For oxalic acid, the proposed model was R(Si)=1.89x10(-12)x[(25x)/(1+25x)]+1.93x10(-12)x[(1990x1)/(1+1990x1)] (R2=0.9763), where x and x1 denote the concentrations of HL(oxalic) and L(oxalic), respectively, and x1=10(-3.81)xx/[H+]. For malic acid, the model was R(Si)=4.79x10(-12)x[(328x)/(1+328x)]+1.67x10(-13)x[(1149x1)/(1+1149x1)] (R2=0.9452), where x and x1 denote the concentrations of HL(malic) and L(malic), respectively, and x1=10(-5.11)xx/[H+], and for citric acid, the model was R(Si)=4.73x10(-12)x[(845x)/(1+845x)]+4.68x10(-12)x[(2855x1)/(1+2855x1)] (R2=0.9682), where x and x1 denote the concentrations of H2L(citric) and L(citric), respectively, and [Formula: see text] .