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It is an increasingly mature application solution that triboelectric nanogenerator (TENG) supplies power to electronic devices through its power management system (PMS). However, the previous PMS is able to manage a limited voltage magnitude and the energy storage elements are limited to capacitors. This work proposes an ultrahigh voltage PMS (UV-PMS) to realize the charging of commercial lithium cells (LCs) by TENG. The design of UV-PMS enables energy management of TENGs with ultrahigh open-circuit voltages up to 3500 V and boosts the peak charging current from 30.9 µA to 2.77 mA, an increase of 89.64 times. With the introduction of UV-PMS, the effective charging capacity of LC charged by a TENG at a working frequency of 1.5 Hz for 1 h comes to 429.7 µAh, making a 75.3 times enhancement compared to charging by TENG directly. The maximum charging power comes to 1.56 mW. The energy storage efficiency is above 97% and the overall charge efficiency can be maintained at 81.2%. This work provides a reliable strategy for TENG to store energy in LC, and has promising applications in energy storage, LC's life, and self-powered systems.
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Secondary cooling electromagnetic stirring (S-EMS) significantly impacts the internal quality of continuous casting slabs. In order to investigate the effects of S-EMS modes on segregation in slabs, a three-dimensional numerical model of the full-scale flow field, solidification, and mass transfer was established. A comparative analysis was conducted between continuous electromagnetic stirring and alternate stirring modes regarding their impacts on steel flow, solidification, and carbon segregation. The results indicated that adopting the alternate stirring mode was more advantageous for achieving uniform flow fields and reducing the disparity in solidification endpoints, thus mitigating carbon segregation. Specifically, the central carbon segregation index under continuous stirring at 320 A was 1.236, with an average of 1.247, while under alternate stirring, the central carbon segregation index decreased to 1.222 with an average of 1.227.
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Recently, discarded electronic products have caused serious environmental pollution and information security issues, which have attracted widespread attention. Here, a degradable tribotronic transistor (DTT) for self-destructing intelligent package e-labels has been developed, integrated by a triboelectric nanogenerator and a protonic field-effect transistor with sodium alginate as a dielectric layer. The triboelectric potential generated by external contact electrification is used as the gate voltage of the organic field-effect transistor, which regulates carrier transport through proton migration/accumulation. The DTT has successfully demonstrated its output characteristics with a high sensitivity of 0.336 mm-1 and a resolution of over 100 µm. Moreover, the DTT can be dissolved in water within 3 min and completely degraded in soil within 12 days, demonstrating its excellent degradation characteristics, which may contribute to environmental protection. Finally, an intelligent package e-label based on the modulation of the DTT is demonstrated, which can display information about the package by a human touch. The e-label will automatically fail due to the degradation of the DTT over time, achieving the purpose of information confidentiality. This work has not only presented a degradable tribotronic transistor for package e-labels but also exhibited bright prospects in military security, information hiding, logistics privacy, and personal affairs.
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Triclocarban (TCC) and triclosan (TCS) have been detected ubiquitously in human body and evoked increasing concerns. This study aimed to reveal the induction risks of TCC and TCS on triple negative breast cancer through non-genomic GPER-mediated signaling pathways. Molecular simulation indicated that TCC exhibited higher GPER binding affinity than TCS theoretically. Calcium mobilization assay displayed that TCC/TCS activated GPER signaling pathway with the lowest observed effective concentrations (LOEC) of 10 nM/100 nM. TCC and TCS also upregulated MMP-2/9, EGFR, MAPK3 but downregulated MAPK8 via GPER-mediated signaling pathway. Proliferation assay showed that TCC/TCS induced 4 T1 breast cancer cells proliferation with the LOEC of 100 nM/1000 nM. Wound-healing and transwell assays showed that TCC/TCS promoted 4 T1 cells migration in a concentration-dependent manner with the LOEC of 10 nM. The effects of TCC on breast cancer cells proliferation and migration were stronger than TCS and both were regulated by GPER. TCC/TCS induced migratory effects were more significantly than proliferative effect. Mechanism study showed that TCC/TCS downregulated the expression of epithelial marker (E-cadherin) but upregulated mesenchymal markers (snail and N-cadherin), which was reversed by GPER inhibitor G15. These biomarkers results indicated that TCC/TCS-induced 4 T1 cells migration was a classic epithelial to mesenchymal transition mechanism regulated by GPER signaling pathway. Orthotopic tumor model verified that TCC promoted breast cancer in-situ tumor growth and distal tissue metastasis via GPER-mediated signaling pathway at human-exposure level of 10 mg/kg/d. TCC-induced tissue metastasis of breast cancer was more significantly than in-situ tumor growth. Overall, we demonstrated for the first time that TCC/TCS could activate the GPER signaling pathways to induce breast cancer progression.
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Neoplasias da Mama , Carbanilidas , Receptores de Estrogênio , Receptores Acoplados a Proteínas G , Transdução de Sinais , Triclosan , Carbanilidas/toxicidade , Transdução de Sinais/efeitos dos fármacos , Triclosan/toxicidade , Humanos , Feminino , Neoplasias da Mama/patologia , Receptores Acoplados a Proteínas G/metabolismo , Receptores de Estrogênio/metabolismo , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Camundongos , Animais , Movimento Celular/efeitos dos fármacosRESUMO
Intramuscular fat (IMF) plays a crucial role in enhancing meat quality, enriching meat flavor, and overall improving palatability. In this study, Single-cell RNA sequencing was employed to analyze the longissimus dorsi (LD) obtained from Guangdong small-ear spotted pigs (GDSS, with high IMF) and Yorkshire pigs (YK, with low IMF). GDSS had significantly more Fibro/Adipogenic Progenitor (FAPs), in which the CD9 negative FAPs (FAPCD9-) having adipogenic potential, as demonstrated by in vitro assays using cells originated from mouse muscle. On the other hand, Yorkshire had more fibro-inflammatory progenitors (FIPs, marked with FAPCD9+), presenting higher expression of the FBN1-Integrin α5ß1. FBN1-Integrin α5ß1 could inhibit insulin signaling in FAPCD9-, suppressing adipogenic differentiation. Our results demonstrated that fat-type pigs possess a greater number of FAPCD9-, which are the exclusive cells in muscle capable of differentiating into adipocytes. Moreover, lean-type pigs exhibit higher expression of FBN1-Integrin α5ß1 axis, which inhibits adipocyte differentiation. These results appropriately explain the observed higher IMF content in fat-type pigs.
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Introduction: This study aims to explore the important factors affecting the characteristics of different parts of pork. Methods: Lipidomics and proteomics methods were used to analyze DAL (differential lipids) and DAPs (differential proteins) in five different parts (longissimus dorsi, belly meat, loin, forelegs and buttocks) of Duhua pig (Duroc × Guangdong small spotted pig), to identify potential pathways affecting meat quality, investigating fat deposition in pork and its lipid-protein interactions. Results: The results show that TG (triglyceride) is the lipid subclass with the highest proportion in muscle, and the pathway with the most significantly enriched lipids is GP. DAP clustered on several GO terms closely related to lipid metabolism and lipogenesis (lipid binding, lipid metabolism, lipid transport, and lipid regulation). In KEGG analysis, there are two main DAP aggregation pathways related to lipid metabolism, namely Fatty acid degradation and oxidative phosphorylation. In PPI analysis, we screened out 31 core proteins, among which NDUFA6, NDUFA9 and ACO2 are the most critical. Discussion: PC (phosphatidylcholine) is regulated by SNX5, THBS1, ANXA7, TPP1, CAVIN2, and VDAC2 in the phospholipid binding pathway. TG is regulated by AUH/HADH/ACADM/ACADL/HADHA in the lipid oxidation and lipid modification pathways. Potential biomarkers are rich in SFA, MUFA and PUFA respectively, the amounts of SFA, MUFA and PUFA in the lipid measurement results are consistent with the up- and down-regulation of potential biomarker lipids. This study clarified the differences in protein and lipid compositions in different parts of Duhua pigs and provided data support for revealing the interactions between pork lipids and proteins. These findings provide contributions to the study of intramuscular fat deposition in pork from a genetic and nutritional perspective.
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In rice-vegetable rotation systems in tropical areas, a large amount of nitrate nitrogen accumulates after fertilization in the melon and vegetable season, which leads to the leaching of nitrate nitrogen and a large amount of N2O emission after the seasonal flooding of rice, which leads to nitrogen loss and intensification of the greenhouse effect. How to improve the utilization rate of nitrate nitrogen and reduce N2O emissions has become an urgent problem to be solved. Six treatments were set up [200 mg·kg-1 KNO3 (CK); 200 mg·kg-1 KNO3 + 2% biochar addition (B); 200 mg·kg-1 KNO3+1% peanut straw addition (P); 200 mg·kg-1 KNO3 + 2% biochar + 1% peanut straw addition (P+B); 200 mg·kg-1 KNO3 + 1% rice straw addition (R); 200 mg·kg-1 KNO3 + 2% biochar+1% rice straw addition (R+B)] and cultured at 25â for 114 d to explore the effects of organic material addition on greenhouse gas emissions and nitrogen use after flooding in high nitrate nitrogen soil. The results showed that compared with that in CK, adding straw or combining straw with biochar significantly increased soil pH (P<0.05). The B and P treatments significantly increased the cumulative N2O emissions by 41.6% and 28.5% (P<0.05), and the P+B, R, and R+B treatments significantly decreased the cumulative N2O emissions by 14.1%, 24.7%, and 36.7% (P<0.05), respectively. The addition of straw increased the net warming potential of greenhouse gases (NGWP). The addition of coir biochar significantly reduced the effect of straw on NGWP (P<0.05). The combined application of straw and biochar decreased NGWP, and P+B significantly decreased NGWP, but that with R+B was not significant (P>0.05). Adding straw or biochar significantly increased soil microbial biomass carbon (MBC) (P<0.05), and that of P+B was the highest (502.26 mg·kg-1). The combined application of straw and biochar increased soil microbial biomass nitrogen (MBN), and that of P+B was the highest. The N2O emission flux was negatively correlated with pH (P<0.01) and positively correlated with NH4+-N and NO3--N (P<0.01). The cumulative emission of N2O was negatively correlated with MBN (P<0.05). There was a significant negative correlation between NO3--N and MBN (P<0.01), indicating that the reduction in NO3--N was likely to be held by microorganisms, and the increase in the microbial hold of NO3--N also reduced N2O emission. In conclusion, the combined application of peanut straw and coconut shell biochar could significantly inhibit N2O emission and increase soil MBC and MBN, which is a reasonable measure to make full use of nitrogen fertilizer, reduce nitrogen loss, and slow down N2O emission after the season of Hainan vegetables.
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Gases de Efeito Estufa , Oryza , Solo/química , Gases de Efeito Estufa/análise , Verduras , Agricultura/métodos , Nitratos , Nitrogênio , Óxido Nitroso/análise , Carvão Vegetal , China , FertilizantesRESUMO
Sesame (Sesamum indicum L.) is a valuable oilseed crop with numerous nutritional benefits containing a diverse range of bioactive compounds. However, sesame is also considered an allergenic food that triggers various mild to severe adverse reactions (e.g., anaphylaxis). Strict dietary avoidance of sesame components is the best option to protect the sensitized consumers. Sesame or sesame-derived foods are always consumed after certain food processing operations, which would cause a considerable impact on the structure of sesame proteins, changing their sensitization capacity and detectability. In the review, the molecular structure properties, and immunological characteristics of the sesame allergens were described. Meanwhile, the influence of food processing techniques on sesame proteins and the relevant detection techniques used for the sesame allergens quantification are also emphasized critically. Hopefully, this review could provide valuable insight into the development and management for the new "Big Eight" sesame allergen in food industry.
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This study aims to overcome the drawbacks associated with hydroxyapatite (HAP) dense structures after sintering, which often result in undesirable features such as large grain size, reduced porosity, high crystallinity, and low specific surface area. These characteristics hinder osseointegration and limit the clinical applicability of the material. To address these issues, a new method involving the preparation of hollow hydroxyapatite (hHAP) microspheres has been proposed. These microspheres exhibit distinctive traits including weak crystallization, high specific surface area, and increased porosity. The weak crystallization aligns more closely with early mineralization products found in the human body and animals. Moreover, the microspheres' high specific surface area and porosity offer advantages for protein loading and facilitating osteoblast attachment. This innovative approach not only mitigates the limitations of conventional HAP structures but also holds the potential for improving the effectiveness of hydroxyapatite in biomedical applications, particularly in enhancing osseointegration. Three-dimensional printed hHAP/chitosan (CS) scaffolds with different hHAP concentration gradients were manufactured, and the physical and biological properties of each group were systematically evaluated. In vitro and in vivo experiments show that the hHAP/CS scaffold has excellent performance in bone remodeling. Furthermore, in-scaffold components, hHAP and CS were cocultured with bone marrow mesenchymal stem cells to explore the regulatory role of hHAP and CS in the process of bone healing and to reveal the cell-level specific regulatory network activated by hHAP. Enrichment analysis showed that hHAP can promote bone regeneration and reconstruction by recruiting calcium ions and regulating inflammatory reactions.
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Quitosana , Durapatita , Animais , Humanos , Durapatita/farmacologia , Durapatita/química , Alicerces Teciduais/química , Cálcio , Osteogênese , Regeneração Óssea/fisiologia , Quitosana/química , Impressão Tridimensional , Porosidade , ÍonsRESUMO
The heat transfer of a slab is significantly influenced by roll contact during the continuous casting process. The influence of roll contact calculation methods on the predicted heat transfer results has not been previously investigated. In this work, the non-uniform solidification of the wide-thick slab was studied with a 2D heat transfer model using real roll contact method (R. method) and equivalent roll contact method (E. method). The predicted slab surface temperature and shell thickness were verified with the measured results of the infrared camera and nail shooting experiments, respectively. Then, the predicted heat transfer results (including the slab surface temperature, mushy region length, and solidification end position) for the wide-thick slab with different thicknesses and different casting speeds were calculated using the E. method and R. method, and the influence of these two roll contact methods on the predicted heat transfer results was discussed for the first time. The results show that both these two roll contact methods could be applied to accurately predict the slab surface temperature without considering the transient temperature dips in the roll-slab contact regions. However, the deviation of the predicted mushy region length and solidification end position using the E. method are obvious. Compared with the R. method, the predicted mushy region length obtained using the E. method is larger and the solidification end obviously subsequently moves along the casting direction.
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The immune microenvironment plays a pivotal role in osteoanagenesis. Biomaterials can modulate osteogenic efficacy by inducing specific local immune reactions. As 3D-printing technology advances, digital light projection printing has emerged as a promising method for creating large scale, high-precision biomaterial scaffolds. By adjusting the solid content and the sintering conditions during printing, the pore size of biomaterials can be meticulously controlled. Yet, the systematic influence of pore size on the immune microenvironment remains uncharted. We fabricated 3D-printed hydroxyapatite bioceramic scaffolds with three distinct pore sizes: 400 µm, 600 µm, and 800 µm. Our study revealed that scaffolds with a pore size of 600 µm promote macrophage M2 polarization, which is achieved by upregulating interferon-beta and HIF-1α production. When these materials were implanted subcutaneously in rats and within rabbit skulls, we observed that the 600 µm scaffolds notably improved the long-term inflammatory response, fostered vascular proliferation, and augmented new bone growth. This research paves the way for innovative therapeutic strategies for treating large segmental bone defects in clinical settings.
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Developing self-powered smart wireless sensor networks by harvesting industrial environmental weak vibration energy remains a challenge and an impending need for enabling the widespread rollout of the industrial internet of things (IIoT). This work reports a self-powered wireless temperature and vibration monitoring system (WTVMS) based on a vibrational triboelectric nanogenerator (V-TENG) and a piezoelectric nanogenerator (PENG) for weak vibration energy collection and information sensing. Therein, the V-TENG can scavenge weak vibration energy down to 80 µm to power the system through a power management module, while the PENG is able to supply the frequency signal to the system by a comparison circuit. In an industrial vibration environment where the vibration frequency and amplitude are 20 Hz and 100 µm, respectively, the WTVMS can upload temperature and frequency information on the equipment to the cloud in combination with the narrowband IoT technology to realize real-time information monitoring. Furthermore, the WTVMS can work continuously for more than 2 months, during which the V-TENG can operate up to 100 million cycles, achieving ultrahigh stability and durability. By integrating weak vibration energy harvesting and active sensing technology, the WTVMS can be used for real-time online monitoring and early fault diagnosis of vibration equipment, which has great application prospects in industrial production, machinery manufacturing, traffic transportation, and intelligent IIoT.
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Introduction: Bone tissue engineering is a promising method to treat bone defects. However, the current methods of preparing composite materials that mimic the complex structure and biological activity of natural bone are challenging for recruitment of bone marrow mesenchymal stem cells (BMSCs), which affects the application of these materials in situ bone regeneration. Hollow hydroxyapatite microspheres (HHMs) possess a natural porous bone structure, good adsorption, and slow release of chemokines, but have low ability to recruit BMSCs and induce osteogenesis. In this study, The HHM/chitosan (CS) and recombinant human C-X-C motif chemokine ligand 13 (rhCXCL13)-HHM/CS biomimetic scaffolds that optimize bone regeneration and investigated their mechanism of BMSC recruitment and osteogenesis through cell and animal experiments and transcriptomic sequencing. Methods: Evaluate the physical characteristics of the HHM/CS and rhCXCL13-HHM/CS biomimetic scaffolds through Scanning Electron Microscopy (SEM), X-Ray Diffraction (XRD), and the cumulative release curve of rhCXCL13. Transwell migration experiments and co-culture with BMSCs were conducted to study the recruitment ability and osteogenic differentiation of the scaffolds. Transcriptomic sequencing was performed to analyze the osteogenic differentiation mechanism. The osteogenesis and bone healing performance were evaluated using a rabbit radial defect model. Results: SEM demonstrated that the rhCXCL13-HHM/CS scaffold comprised hydroxyapatite microspheres in a porous three-dimensional network. The rhCXCL13 showed excellent sustained release capability. The rhCXCL13-HHM/CS scaffold could recruit BMSCs and induce bone regeneration. Transcriptome sequencing and experimental results showed that the osteogenesis mechanism of rhCXCL13-HHM/CS was through the PI3K-AKT pathway. In vivo, the rhCXCL13-HHM/CS scaffold significantly promoted osteogenesis and angiogenesis at 12 weeks after surgery. Conclusion: The rhCXCL13-HHM/CS scaffold demonstrates excellent potential for BMSC recruitment, osteogenesis, vascularized tissue-engineered bone reconstruction, and drug delivery, providing a theoretical basis for material osteogenesis mechanism study and promising clinical applications for treating large bone defects.
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Quitosana , Osteogênese , Animais , Humanos , Coelhos , Durapatita/farmacologia , Durapatita/química , Alicerces Teciduais/química , Microesferas , Ligantes , Fosfatidilinositol 3-Quinases , Regeneração Óssea , Engenharia Tecidual/métodos , Diferenciação CelularRESUMO
Sound monitoring has been widely used in the field of the Internet of Things (IoT), in which the sensors are mainly powered by batteries with high power consumption and limited life. Here, a near-zero quiescent power sound wake-up and identification system based on a triboelectric nanogenerator (TENG) is proposed, in which the sound TENG (S-TENG) is used for ambient sound energy harvesting and system activation. Once the sound intensity is higher than 65 dB, the converted and stored electric energy by the S-TENG can wake up the system within 0.5 s. By integrating a deep learning technique, the system is used for identifying sound sources, such as drilling, child playing, dog barking, and street music. In the active mode, the sound signals are recorded by a microelectromechanical systems (MEMS) microphone and then sent to a remote computer for sound recognition through a wireless transmitter within 2.8 s. In the standby mode, the ambient sound is not enough to wake up the system, and the quiescent power consumption is only 55 nW. This work provides a triboelectric sensor-based ultralow quiescent power sound wake-up system, which has shown excellent application prospects in smart homes, unmanned monitoring, and the Internet of Things.
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Coal bed methane drainage is the main approach to lower risks of coal seam while raising the efficiency in natural resource utilization. The negative pressure used for extraction in coal mines is largely determined empirically due to a lack of experimental research on how coal permeability changes under the combined influence of effective stress and negative pressure. This results in low gas extraction efficiency and concentration. In this paper, to study the effect law of complex stress and extraction on coal permeability during coal and gas co-mining, a test system was specially designed to determine the gas flow and coal permeability of coal samples under different stress paths and negative pressure conditions in the lab. The study analyzed the correlation between coal permeability, effective stress, and negative pressure and subsequently developed a permeability evolution model for gas-bearing coal under negative pressure conditions. The results showed that the permeability of coal increases with the increase in negative pressure and decreases with the increase in effective stress; the permeability of coal can be abruptly changed by changes in stress loading patterns; the established model of permeability evolution of gas-bearing coal can better reflect the correlation between permeability, effective stress, and negative pressure. The research outcomes offer a valuable theoretical foundation for the efficient extraction and utilization of methane in coal mines.
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Background: Cervical cancer (CC) is a major health threat to females, and distal metastasis is common in patients with advanced CC. Anoikis is necessary for the development of distal metastases. Understanding the mechanisms associated with anoikis in CC is essential to improve its survival rate. Methods: The expression matrix of long non-coding RNAs (lncRNAs) from cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC) patients was extracted from The Cancer Genome Atlas (TCGA), and highly relevant anoikis-related lncRNAs (ARLs) were identified by the single sample gene set enrichment analysis (ssGSEA) method. ARLs-related molecular subtypes were discerned based on prognosis-related ARLs. ARLs-related prognostic risk score (APR_Score) was calculated and risk model was constructed using LASSO COX and COX models. In addition, we also assessed immune cell activity in the immune microenvironment (TME) for both subtypes and APR_Score groups. A nomogram was utilized for predicting improved clinical outcome. Finally, this study also discussed the potential of ARLs-related signatures in predicting response to immunotherapy and small molecular drugs. Results: Three ARLs-related subtypes were identified from TCGA-CESC (AC1, AC2, and AC3), with AC3 patients having the highest ARG scores, higher angiogenesis scores, and the worst prognosis. AC3 had lower immune cell scores in TME but higher immune checkpoint gene expression and higher potential for immune escape. Next, we constructed a prognostic risk model consisting of 7-ARLs. The APR_Score exhibited a greater robustness as an independent prognostic indicator in predicting prognosis, and the nomogram was a valuable tool for survival prediction. ARLs-related signatures emerged as a potential novel indicator for immunotherapy and small molecular drug selection. Conclusion: We firstly constructed novel ARLs-related signatures capable of predicting prognosis and offered novel ideas for therapy response in CC patients.
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Renal fibrosis is a common pathological outcome of various chronic kidney diseases, and as yet, there is no specific treatment. Dapagliflozin has shown renal protection in some clinical trials as a glucose-lowering drug, but its role and mechanism on renal fibrosis remain unclear. In this study, we used a 0.2% adenine diet-induced renal fibrosis mouse model to investigate whether dapagliflozin could protect renal function and alleviate renal fibrosis in this animal model. In vivo, we found that dapagliflozin's protective effect on renal fibrosis was associated with 1) sustaining mitochondrial integrity and respiratory chain complex expression, maintained the amount of mitochondria; 2) improving fatty acid oxidation level with increased expression of CPT1-α, PPAR-α, ACOX1, and ACOX2; 3) reducing inflammation and oxidative stress, likely via regulation of IL-1ß, IL-6, TNF-α, MCP-1, cxcl-1 expression, and glutathione (GSH) activity, superoxide dismutase (SOD) and malondialdehyde (MDA) levels; and 4) inhibiting the activation of the TGF-ß1/MAPK pathway. In HK2 cells treated with TGF-ß1, dapagliflozin reduced the expression of FN and α-SMA, improved mitochondrial respiratory chain complex expression, and inhibited activation of the TGF-ß1/MAPK pathway.
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Despite the fact that tropomyosin (TM) has highly stable structural characteristics, thermal processing can adversely influence its immunodetection, and the mechanism involved has not been elucidated. Purified TM was heated at various temperatures, and then the IgG/IgE-binding capacity and immunodetection recovery were determined; meanwhile, the structural alterations were analyzed via spectroscopic and molecular dynamics simulation techniques. The obtained results demonstrated that heat-treated TM showed significantly increased IgG/IgE reactivity, confirmed by indirect ELISA and immunoblotting analysis, which might be attributed to the increased structural flexibility, and thus allowed TM to be recognized IgG/IgE easily. However, these structural alterations during thermal processing would contribute to the masking of some epitopes located in TM's surface due to the presence of curled or folded conformation with a considerable reduction of the solvent-accessible surface and radius of gyration, which primarily caused immunodetection recovery reduction in the sandwich ELISA (sELISA) test. The number of antigen binding sites might play a crucial role in a sandwich immunodetection system for sensitive and precise analysis in processed foods.
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Cow milk protein allergy (CMPA) induced by casein poses major health challenges that hinders the consumption of milk-based formulas. In this study, a novel sequential enzymatic hydrolysis catalysed by chymosin and papain was proposed to reduce casein antigenicity. Its effects on reducing casein antigenicity, structural properties and peptide profiles were evaluated by ELISA, multispectral techniques and peptidome analysis. It was revealed that the sequential enzymatic hydrolysis obtained a similarly residual antigenicity level in a shorter time (60 min) compared to papain-hydrolysis for 360 min. The hydrolysis-site at Tyr residues accessibility of papain was increased to 36.84 % by the chymosin pretreatment and it was significantly higher than 26.93 % obtained by only papain for 60 min. Moreover, the sequential enzymatic hydrolysis led to decrease in the large fragment peptides from αs1 casein. These findings suggested that the proposed sequential enzymatic hydrolysis can be exploited in the development of CMPA-free formulas.
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Quimosina , Hipersensibilidade a Leite , Animais , Bovinos , Feminino , Caseínas , Papaína , Hidrólise , TirosinaRESUMO
αs1-Casein (αs1-CN) is a major cow milk allergen, while the tertiary structure of αs1-CN and conformational epitopes of αs1-CN have not been clarified. Here, a reasonable three-dimensional structure of αs1-CN was established using ab initio methods, and hot-spot residues and epitopes were investigated by combining molecular dynamics simulation, peptides synthesis, and ELISA. Obtained results demonstrated that the binding mechanism between αs1-CN and IgG was located on three main regions: a helical structure zone (E77-Q97), the flexible loop zone (Y154-T174), and a flexible C-terminal (N190-L198), mainly connecting via hydrogen bond and ionic bonds. The hydrolysates produced by papain with lowest antigenicity (12.43%), which could considerably destroy the essential epitopes of αs1-CN confirmed by epitope synthesis, and LC-MS/MS. The results reported herein would provide novel insights into the interface interactions between αs1-CN and IgG, and prove valuable for developing hypoallergenic infant-formula and peptide vaccines for allergen-specific immunotherapy.