Comprehensive transcriptomic and metabolomic analysis of porcine intestinal epithelial cells after PDCoV infection.

Introduction: Porcine deltacoronavirus (PDCoV), an emerging swine enteropathogenic coronavirus with worldwide distribution, mainly infects newborn piglets with severe diarrhea, vomiting, dehydration, and even death, causing huge economic losses to the pig industry. However, the underlying pathogenic mechanisms of PDCoV infection and the effects of PDCoV infection on host transcripts and metabolites remain incompletely understood. Methods: This study investigated a combined transcriptomic and metabolomic analysis of porcine intestinal epithelial cells (IPEC-J2) following PDCoV infection by LC/MS and RNA-seq techniques. A total of 1,401 differentially expressed genes and 254 differentially accumulated metabolites were detected in the comparison group of PDCoV-infected vs. mock-infected. Results and discussion: We found that PDCoV infection regulates gene sets associated with multiple signaling pathways, including the neuroactive ligand-receptor interaction, cytokine-cytokine receptor interaction, MAPK signaling pathway, chemokine signaling pathway, ras signaling pathway and so on. Besides, the metabolomic results showed that biosynthesis of cofactors, nucleotide metabolism, protein digestion and absorption, and biosynthesis of amino acid were involved in PDCoV infection. Moreover, integrated transcriptomics and metabolomics analyses revealed the involvement of ferroptosis in PDCoV infection, and exogenous addition of the ferroptosis activator erastin significantly inhibited PDCoV replication. Overall, these unique transcriptional and metabolic reprogramming features may provide a better understanding of PDCoV-infected IPEC-J2 cells and potential targets for antiviral treatment.

Melatonin Modulates Tomato Root Morphology by Regulating Key Genes and Endogenous Hormones.

Melatonin plays a vital role in plant growth and development. In this study, we treated hydroponically grown tomato roots with various concentrations of exogenous melatonin (0, 10, 30, and 50 µmol·L-1). We utilized root scanning and microscopy to examine alterations in root morphology and cell differentiation and elucidated the mechanism by which melatonin regulates these changes through the interplay with endogenous hormones and relevant genes. The results showed that for melatonin at concentrations ranging between 10 and 30 µmol·L-1, the development of lateral roots were significantly stimulated, the root hair growth was enhanced, and biomass accumulation and root activity were increased. Furthermore, we elucidated that melatonin acts as a mediator for the expression of genes, such as SlCDKA1, SlCYCA3;1, SlARF2, SlF3H, and SlKT1, which are involved in the regulation of root morphology changes. Additionally, we observed that melatonin influences the levels of endogenous hormones, including ZT, GA3, IAA, ABA, and BR, which subsequently impact the root morphology development of tomato roots. In summary, this study shows that tomato root morphology can be promoted by the optimal concentration of exogenous melatonin (10-30 µmol·L-1).

Lateral Habenula Neurons Signal Cold Aversion and Participate in Cold Aversion.

The aversion to cold is a fundamental motivated behavior that contributes to the body temperature homeostasis. However, the involvement of the lateral habenula (LHb) as a regulatory hub for negative emotions in this physiological process remains uninvestigated. In this study, we demonstrate an elevation in the population activity of LHb neurons following exposure to cold stimuli. Additionally, we establish the necessity of Vglut2-expressing neurons within the LHb for the encoding of cold aversion behaviors. Furthermore, we have elucidated a neural circuit from excitatory neurons of the dorsomedial hypothalamus (DMH) to LHb that plays a crucial role in this progress. Manipulation of the DMH-LHb circuit has a significant impact on cold aversion behavior in mice. It is worth noting that this circuit does not exhibit any noticeable effects on autonomic thermoregulation or depression-like behavior. The identification of these neural mechanisms involved in behavioral thermoregulation provides a promising avenue for future research.

Integrated metabolomics and transcriptomics analyses reveal metabolic responses to TGEV infection in porcine intestinal epithelial cells.

Transmissible gastroenteritis virus (TGEV) is a coronavirus that infects piglets with severe diarrhoea, vomiting, dehydration, and even death, causing huge economic losses to the pig industry. The underlying pathogenesis of TGEV infection and the effects of TGEV infection on host metabolites remain poorly understood. To investigate the critical metabolites and regulatory factors during TGEV infection in intestinal porcine epithelial cells (IPEC-J2), we performed metabolomic and transcriptomic analyses of TGEV-infected IPEC-J2 cells by LC/MS and RNA-seq techniques. A total of 87 differential metabolites and 489 differentially expressed genes were detected. A series of metabolites and candidate genes from glutathione metabolism and AMPK signalling pathway were examined through combined analysis of metabolome and transcriptome. We found glutathione peroxidase 3 (GPX3) is markedly reduced after TGEV infection, and a significant negative correlation between AMPK signalling pathway and TGEV infection. Exogenous addition of the AMPK activator COH-SR4 significantly downregulates stearoyl coenzyme A (SCD1) mRNA and inhibits TGEV replication; while exogenous GSK-690693 significantly promotes TGEV infection by inhibiting AMPK signalling pathway. In summary, our study provides insights into the key metabolites and regulators for TGEV infection from the metabolome and transcriptome perspective, which will offer promising antiviral metabolic and molecular targets and enrich the understanding of the existence of a similar mechanism in the host.

Integrated of multi-omics and molecular docking reveal PHGDH, PSAT1 and PSPH in the serine synthetic pathway as potential targets of T-2 toxin exposure in pig intestinal tract.

T-2 toxin (T-2) with a molecular weight of 466.52 g/mol is an inevitable mycotoxin in food products and feeds, posing a significant threat to human and animal health. However, the underlying molecular mechanisms of the cytotoxic effects of T-2 exposure on porcine intestinal epithelial cells (IPEC-J2) remain unclear. Here, we investigated the cytotoxic effects of T-2 exposure on IPEC-J2 through the detection of cell viability, cell morphology, mitochondrial membrane potential, ROS, apoptosis and autophagy. Further transcriptomic and proteomic analyses of IPEC-J2 upon T-2 exposure were performed by using RNA-seq and TMT techniques. A total of 546 differential expressed genes (DEGs) and 269 differentially expressed proteins (DEPs) were detected. Among these, 24 common DEGs/DEPs were involved in IPEC-J2 upon T-2 exposure. Interestingly, molecular docking analysis revealed potential interactions between T-2 and three key enzymes (PHGDP, PSAT1, and PSPH) in the serine biosynthesis pathway. Besides, further experimental showed that PSAT1 knockdown exacerbated T-2-induced oxidative damage. Together, our findings indicated that the serine biosynthesis pathway including PHGDP, PSAT1, PSPH genes probably acts critical roles in the regulation of T-2-induced cell damage. This study provided new insights into the global molecular effects of T-2 exposure and identified the serine biosynthesis pathway as molecular targets and potential treatment strategies against T-2.

Porcine Deltacoronavirus Infection Disrupts the Intestinal Mucosal Barrier and Inhibits Intestinal Stem Cell Differentiation to Goblet Cells via the Notch Signaling Pathway.

Goblet cells and their secreted mucus are important elements of the intestinal mucosal barrier, which allows host cells to resist invasion by intestinal pathogens. Porcine deltacoronavirus (PDCoV) is an emerging swine enteric virus that causes severe diarrhea in pigs and causes large economic losses to pork producers worldwide. To date, the molecular mechanisms by which PDCoV regulates the function and differentiation of goblet cells and disrupts the intestinal mucosal barrier remain to be determined. Here, we report that in newborn piglets, PDCoV infection disrupts the intestinal barrier: specifically, there is intestinal villus atrophy, crypt depth increases, and tight junctions are disrupted. There is also a significant reduction in the number of goblet cells and the expression of MUC-2. In vitro, using intestinal monolayer organoids, we found that PDCoV infection activates the Notch signaling pathway, resulting in upregulated expression of HES-1 and downregulated expression of ATOH-1 and thereby inhibiting the differentiation of intestinal stem cells into goblet cells. Our study shows that PDCoV infection activates the Notch signaling pathway to inhibit the differentiation of goblet cells and their mucus secretion, resulting in disruption of the intestinal mucosal barrier. IMPORTANCE The intestinal mucosal barrier, mainly secreted by the intestinal goblet cells, is a crucial first line of defense against pathogenic microorganisms. PDCoV regulates the function and differentiation of goblet cells, thereby disrupting the mucosal barrier; however, the mechanism by which PDCoV disrupts the barrier is not known. Here, we report that in vivo, PDCoV infection decreases villus length, increases crypt depth, and disrupts tight junctions. Moreover, PDCoV activates the Notch signaling pathway, inhibiting goblet cell differentiation and mucus secretion in vivo and in vitro. Thus, our results provide a novel insight into the mechanism underlying intestinal mucosal barrier dysfunction caused by coronavirus infection.

Study on Multi-Mode Switching Control Strategy of Active Suspension Based on Road Estimation.

In this paper, the least squares method is used to determine the vertical height of the road space domain. Based on the road estimation method, the active suspension control mode switching model is constructed, and the dynamic characteristics of the vehicle in comfort, safety, and integrated modes are analyzed. The vibration signal is collected by the sensor, and the parameters such as vehicle driving conditions are solved for in reverse. A control strategy for multiple mode switching under different road surfaces and speeds is constructed. At the same time, the particle swarm optimization algorithm (PSO) is used to optimize the weight coefficients of LQR control under different modes, and the dynamic performance of vehicle driving is comprehensively analyzed. The test and simulation results show that the road estimation results under different speeds in the same road section are very close to the results obtained by the detection ruler method, and the overall error is less than 2%. Compared with the active suspension controlled by passive and traditional LQR, the multi-mode switching strategy can achieve a better balance between driving comfort and handling safety and stability, and also improve the driving experience more intelligently and comprehensively.

Synthesis of AgNPs from waste mobile phone circuit boards using an emulsion liquid membrane method.

The disposal of mobile phone print circuit boards (WMPCBs) has now been a problem because of the continually increasing production of mobile phones. It could be considered a new source of precious metal but may jeopardize the environment and human health without proper management. An emulsion liquid membrane (ELM) method is applied and improved to recover silver from WMPCBs and prepare AgNPs. The effects of significant operating parameters were studied for a better understanding of the ELM process. Under the optimized conditions i.e., 0.25 % (v/v) Cyanex 302, 6 % (v/v) Span 80, 5 % (w/v) ASC, 3500 rpm emulsification speed, 10 min emulsification time, 300 rpm extraction stirring speed, 10 min extraction stirring time and 3 (v/v) phase ratio, 98.7 % silver was extracted. AgNPs produced were flower structures assembled by a group of individual nanosheets with a diameter of 800-1400 nm and a thickness of 10-14 nm. The purity was 98.7 % and the yield was 96.6 %. AgNPs of oblate spheroidal structure with a diameter of 30-50 nm could be synthesized by controlling the concentration of Span 80 in ELM. The current study intends to fulfill the gap by exploring the recovery of silver from WMPCBs by the ELM method.

Enhancing the technofunctionality of γ-aminobutyric acid enriched germinated wheat by modification of arabinoxylan, gluten proteins and liquid lamella of dough.

To enhance the technofunctionality of germinated wheat enriched with γ-aminobutyric acid, xylanase (Xyn) and glucose oxidase (Gox) were incorporated with emphasis on modifying the key components. Combination of Xyn and Gox enhanced steamed bread quality with optimum loaf volume and textural property. Continuous and dense gluten network was facilitated and improved viscoelasticity of dough. Water solubility of arabinoxylan (AX) enhanced with Xyn and the molecular weight was more homogeneous distributed throughout bread making process with Xyn and Gox. Polymerization behavior of α-/γ-gliadin and glutenin was suppressed in steamed bread, while incorporation of AX to insoluble proteins was enhanced by enzymes. In addition, the promoted formation of high molecular weight glycoprotein in the liquid lamella of dough enhanced the thermal stability of foams and contribute to superior quality of steamed bread. Results demonstrated that germinated wheat could be exploited as a functional ingredient with desirable technofunctionality by modification of the components.

Application of Exogenous Melatonin Improves Tomato Fruit Quality by Promoting the Accumulation of Primary and Secondary Metabolites.

Melatonin plays key roles in improving fruit quality and yield by regulating various aspects of plant growth. However, the effects of how melatonin regulates primary and secondary metabolites during fruit growth and development are poorly understood. In this study, the surfaces of tomato fruit were sprayed with different concentrations of melatonin (0, 50, and 100 µmol·L-1) on the 20th day after anthesis; we used high-performance liquid chromatography (HPLC) and liquid chromatography/mass spectrometry (LC/MS) to determine the changes in primary and secondary metabolite contents during fruit development and measured the activity of sucrose metabolizing enzymes during fruit development. Our results showed that 100 µmol·L-1 melatonin significantly promoted the accumulation of soluble sugar in tomato fruit by increasing the activities of sucrose synthase (SS), sucrose phosphate synthase (SPS), and acid convertase (AI). The application of 100 µmol·L-1 melatonin also increased the contents of ten amino acids in tomato fruit as well as decreased the contents of organic acids. In addition, 100 µmol·L-1 melatonin application also increased the accumulation of some secondary metabolites, such as six phenolic acids, three flavonoids, and volatile substances (including alcohols, aldehydes, and ketones). In conclusion, melatonin application improves the internal nutritional and flavor quality of tomato fruit by regulating the accumulation of primary and secondary metabolites during tomato fruit ripening. In the future, we need to further understand the molecular mechanism of melatonin in tomato fruit to lay a solid foundation for quality improvement breeding.

Impact of Wheat Arabinoxylan with Defined Substitution Patterns on the Heat-Induced Polymerization Behavior of Gluten.

To further depict the interaction mechanism of wheat arabinoxylan (AX) and gluten proteins upon thermal processing, AX was enzymatically tailored with defined substitution patterns and the impact on the heat-induced polymerization behavior of gluten was comparatively studied. The results showed that tailormade AX promoted the formation of glutenin-glutenin and glutenin-gliadin macrocrosslinks upon heating, with the optimal effect detected for AX depleted of Araf of disubstituted Xylp. The tailormade AX, especially AX depleted of monosubstituted Xylp, facilitated the polymerization ability of α-gliadin into glutenin compared with untailored AX. The unfolding process of gluten was partially impeded by AX upon heating, while the tailormade AX promoted the unfolding process. AX could bury Trp and Tyr upon polymerization of glutenin and gliadin and induced the change of the disulfide bridge conformation to a less-stable state, while the effect was alleviated with tailormade AX. The enhanced polymerization with tailormade AX strengthened the gluten network and induced more heterogeneously distributed large protein aggregates.

The impact of subsidy and preferential tax policies on mobile phone recycling: A system dynamics model analysis.

This paper analyzes the impact of possible subsidy and preferential tax policies on mobile phone (MP) recycling industries in China using system dynamics methodology, aiming to offer a reference for the construction of a reasonable policy system for MP recycling. First, different subsidy and preferential tax policies were introduced to establish a system dynamics model. Then the impact of policies proposed on MP recycling industries was analyzed using the model. The results showed that: (1) newly discarded MPs could be completely recycled annually if recyclers could achieve a RMB10 recycling subsidy for each MP formally recycled; (2) an initial subsidy policy can effectively increase the number of remanufacturers, but is harmful to remanufacturing capacity; (3) stage R&D subsidy policy outperforms equalization R&D subsidy policy. Additionally, more subsidies allocated to remanufactures means more profit when R&D subsidy is limited; (4) more tax benefits provide more output and profit. Remanufacturers are more sensitive to tax benefits than recovery enterprises in the short run. Finally, several suggestions related to the construction of MP recycling policy systems were put forward based on evaluation results.

Physicochemical, Nutritional Properties and Metabolomics Analysis Fat Deposition Mechanism of Chahua Chicken No. 2 and Yao Chicken.

Poultry is an important dietary source of animal protein, accounting for approximately 30% of global meat consumption. Because of its low price, low fat and cholesterol content, and no religious restrictions, chicken is considered a widely available healthy meat. Chahua chicken No. 2 is a synthetic breed of Chahua chicken derived from five generations of specialized strain breeding. In this study, Chahua chicken No. 2 (CH) and Yao chicken (Y) were used as the research objects to compare the differences in physicochemical and nutritional indicators of meat quality between the two chicken breeds, and metabolomics was used to analyze the differences in metabolites and lipid metabolism pathways and to explore the expression of genes involved in adipogenesis. The physical index and nutritional value of CH are better than that of Y, and the chemical index of Y is better than that of CH. However, the chemical index results of CH are also within the normal theoretical value range. Comprehensive comparison shows that the meat quality of CH is relatively good. Metabolomics analysis showed that CH and Y had 85 different metabolites, and the differential metabolites were mainly classified into eight categories. KEGG pathway enrichment analysis revealed 13 different metabolic pathways. The screened PPARG, FABP3, ACSL5, FASN, UCP3 and SC5D were negatively correlated with muscle fat deposition, while PPARα, ACACA and ACOX1 were positively correlated with muscle fat deposition. The meat quality of CH was better than Y. The metabolites and metabolic pathways obtained by metabonomics analysis mainly involved the metabolism of amino acids and fatty acids, which were consistent with the differences in meat quality between the two breeds and the contents of precursors affecting flavor. The screened genes were associated with fatty deposition in poultry.

Association between H19 polymorphisms and NSCLC risk in a Chinese Population.

PURPOSE: Long non-coding RNAs (lncRNA) polymorphisms have been reported to associate with the carcinogenicity mechanisms. The association between lncRNA H19 polymorphisms and the risk of non-small cell lung cancer (NSCLC) in Chinese population has not been reported yet. We designed this case-control study to evaluate the effects of H19 polymorphisms on NSCLC susceptibility. METHODS: In this case-control study, four single nucleotide polymorphisms (SNPs) (rs2067051, rs217727, rs2839698 and rs4929984) in H19 gene were genotyped in a Chinese population which consisted of 564 NSCLC cases and 1536 controls. RESULTS: rs2067051 was associated with a significantly decreased risk of NSCLC in our population [AA vs. GG: adjusted odds ratio (OR) = 0.75, 95% confidence interval (CI)=0.60-0.93; Additive model: OR=0.79, 95%CI=0.67-0.93)]. rs217727 was related to significantly increased NSCLC susceptibility (TT vs. CC: OR=1.16, 95%CI=1.01-1.33; Additive model: OR=1.16, 95%CI=1.01-1.33). However, no significant association was observed between rs2839698, rs4929984 and NSCLC risk. CONCLUSIONS: H19 polymorphism rs2067051 and rs217727 might influence NSCLC susceptibility and the mechanism warrants further exploration.