Regulatory mechanism of miR-20a-5p in neuronal damage and inflammation in lipopolysaccharide-induced BV2 cells and MPTP-HCl-induced Parkinson's disease mice.

BACKGROUND: Parkinson's disease (PD) is a prevailing neurodegenerative disorder increasingly affecting the elderly population. The involvement of microRNAs (miRNAs) in PD has been confirmed. We sought to explore the molecular mechanism of miR-20a-5p in PD. METHODS: Lipopolysaccharide (LPS)-induced BV2 cell model and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride (MPTP-HCl)-induced PD mouse model were established. miR-20a-5p, inducible nitric oxide synthase (iNOS), interleukin (IL)-6, tumour necrosis factor (TNF)-α, transforming growth factor (TGF)-ß1, and IL-10 expression in BV2 cells was examined by reverse transcription - quantitative polymerase chain reaction. Cell viability was assessed by MTT assay. The apoptotic rate and levels of Bcl-2, Bax, cleaved caspase-3, and signal transducer and activator of transmission (STAT)3 were examined by flow cytometry and Western blot. Bioinformatics software predicted the potential binding sites of miR-20a-5p and STAT3. Dual-luciferase experiment verified the binding relationship. Iba1-positive and tyrosine hydroxylase (TH)-positive cell numbers in substantia nigra pars compacta were detected by immunohistochemistry. The effect of miR-20a-5p on motor function in MPTP-induced PD mice was detected by Rota-rod test, Pole test, Traction test and Beam-crossing task. RESULTS: miR-20a-5p was under-expressed in LPS-induced BV2 cells. Overexpression of miR-20a-5p increased the viability of LPS-induced BV2 cells and reduced apoptosis rates. Moreover, overexpression of miR-20a-5p reduced cleaved caspase-3, Bax, iNOS, IL-6, and TNF-α and increased Bcl-2 and TGF-ß1, and IL-10. miR-20a-5p targeted STAT3. STAT3 overexpression partially reversed miR-20a-5p overexpression-mediated effects on LPS-induced BV2 cell viability, apoptosis, and inflammatory responses. miR-20a-5p overexpression inhibited MPTP-induced STAT3 and α-synuclein levels, microglia activation, and inflammatory response, and reduced the loss of TH-positive cells in mice. miR-20a-5p overexpression ameliorated MPTP-induced dyskinesia in PD model mice. CONCLUSION: miR-20a-5p alleviates neuronal damage and suppresses inflammation by targeting STAT3 in PD.

Food Hydrocolloids: Structure, Properties, and Applications.

Hydrocolloids are extensively used in the food industry for various functions, including gelling, thickening, stabilizing foams, emulsions, and dispersions, as well as facilitating the controlled release of flavor [...].

Identification of DLEU2 as biomarker based on lncRNA-Mediated ceRNA network in rupture of intracranial aneurysm.

Intracranial aneurysms (IA) is a potentially devastating clinical problem that may cause lethal subarachnoid hemorrhage upon rupture, but the molecular mechanisms remain further elucidated. Our goal in this work was to build the lncRNA-mediated ceRNA network in IS and explore the associated pathways and functions. The deep transcriptome sequencing dataset profile of rupture of IA and normal tissues (SRP150595) was obtained from NCBI database. To determine which genes were differently expressed, weighted gene co-expression network analysis and other integrated bioinformatics techniques were used (DEGs). The action mechanism and associated pathways of DEGs in IA were investigated using GO annotations and KEGG analysis. The Starbase database was used to build the ceRNA network. Vascular smooth muscle cells (VSMC) were used for the transwell assay and CCK-8. A total of 248 common differentially expressed-protein coding RNA and 76 DE-lncRNAs were obtained. Functional enrichment analysis indicated that the DEGs of IA are involved in pathways of inflammation and immune response. A lncRNAs-mediated ceRNA network including lncRNAs BASP1-AS1, DLEU2, LINC02035, LINC02363, MMP25-AS1, AC008771.1 was constructed. The biological behavior of VSMC was suppressed when DLEU2 was knocked down. In conclusion, a lncRNAs-mediated ceRNA network was constructed in IA based on the integrated bioinformatics analyses, in which DLEU2 was identified to be a novel and potential biomarker of IA.

Preparation of Bovine Hides Gelatin by Ultra-High Pressure Technique and the Effect of Its Replacement Fat on the Quality and In Vitro Digestion of Beef Patties.

Beef skin gelatin can be used as a good substitute for animal fat in meat patties. In this paper, the effect of different parameters on low-fat beef patties with cowhide gelatin substituted for beef fat (0, 25%, 50%, 75%, 100%) prepared by ultra-high pressure assisted technology was investigated by texture, cooking loss, and sensory scores. The beef patties were also stored at 0-4 °C for 0, 7, 14, 21, and 28 d. The differences and changing rules of fatty acid and amino acid compositions and contents of beef patties with different fat contents were investigated by simulating gastrointestinal digestion in vitro. The optimal process formulation of low-fat beef patties with cowhide gelatin was determined by experimental optimization as follows: ultra-high pressure 360 MPa, ultra-high of pressure time of 21 min, NaCl addition of 1.5%, compound phosphate addition of 0.3%. The addition of cowhide gelatin significantly increased monounsaturated fatty acids, polyunsaturated fatty acids, amino acid content, and protein digestibility of beef patties (p < 0.05). Moreover, with the extension of storage time, the content of saturated fatty acids was significantly higher (p < 0.05), the content of monounsaturated and polyunsaturated fatty acids was significantly lower (p < 0.05), the content of amino acids was significantly lower (p < 0.05), and protein digestibility was significantly lower (p < 0.05) under all substitution ratios. Overall, beef patties with 75% and 100% substitution ratios had better digestibility characteristics. The results of this study provide a theoretical basis for gelatin's potential as a fat substitute for beef patties and for improving the quality of low-fat meat products.

Mechanism of RNA m6 A methylation modification regulating NLRP3 inflammasome activation for hand, foot, and mouth disease progression.

Some children infected with hand, foot, and mouth disease (HFMD) caused by enterovirus 71 (EV71) progressed to severe disease with various neurological complications in the short term, with a poor prognosis and high mortality. Studies had revealed that RNA N6 -methyladenosine (m6 A) modification had a significant impact on EV71 replication, but it was unknown how m6 A modification regulated the host cell's innate immune response brought on by EV71 infection. We used MeRIP-seq (methylation RNA immunoprecipitation sequencing), RNA-seq (RNA sequencing), cell transfection, and other techniques. MeRIP-seq and RNA-seq results showed the m6 A methylation modification map of control and EV71-infected groups of RD cells. And multilevel validation indicated that decreased expression of demethylase FTO (fat mass and obesity-associated protein) was responsible for the elevated total m6 A modification levels in EV71-infected RD cells and that thioredoxin interacting protein (TXNIP) may be a target gene for demethylase FTO action. Further functional experiments showed that demethylase knockdown of FTO promoted TXNIP expression, activation of NLRP3 inflammasome and promoted the release of proinflammatory factors in vitro, and the opposite result occurred with demethylase FTO overexpression. And further tested in an animal model of EV71 infection in vitro, with results consistent with in vitro. Our findings elucidated that depletion of the demethylase FTO during EV71 infection increased the m6 A modification level of TXNIP mRNA 3' untranslated region (UTR), enhancing mRNA stability, and promoting TXNIP expression. Consequently, the NLRP3 inflammasome was stimulated, leading to the release of proinflammatory factors and facilitating HFMD progression.

Ultra-high pressure-assisted preparation of cowhide gelatin as a promising fat substitute: Improve the nutrition ratio and antioxidant capacity of beef patties.

In this study, low-fat beef patties were prepared by replacing different proportions of beef fat (0, 25, 50, 75, and 100%) with ultra-high pressure-assisted cowhide gelatin. The quality characteristics, lipid oxidation, protein oxidation, and fatty acid profile of beef patties during cold storage at 4 ℃ (0, 7, 14, 21, and 28 days) were evaluated. The results showed that the addition of cowhide gelatin increased the protein content and reduced the fat content of beef patties. The saturated fatty acid (SFA) content of beef patties significantly reduced and the polyunsaturated fatty acid (PUFA) content significantly increased with the increase in the replacement ratio. Especially, the 100% substitution group had the highest PUFA content; the SFA content and the n-6/n-3 ratio could be reduced by about 56% and 31%, respectively. During the cold storage period, the moisture content of beef patties increased significantly, while the cooking loss was the opposite, with the increase in the fat replacement rate. The L* and a* values of raw patties prepared with gelatin were significantly higher than those in the control group. Furthermore, adding cowhide gelatin significantly reduced the thiobarbituric acid, peroxide, and carbonyl contents of patties, but increased their total sulfhydryl content throughout the refrigeration period. Thus, it was concluded that cowhide-based gelatin was a promising alternative to replace beef fat in low-fat beef patties.

Effect of Salt (Sodium Chloride) Replacement With Potassium Chloride, High Pressure Processing, and Cold Storage at 4°C on Beef Sausage Volatile Compounds.

This study aimed to investigate the partial substitution of 0, 25 and 50% sodium chloride (NaCl) by potassium chloride (KCl) coupled with high-pressure processing (HPP) effects on volatile compounds and lipid oxidation of beef sausage at five (0, 3, 7, 14, and 21) different cold storage days (4°C). The odor activity values (OAV) of the various compounds were visualized by heat map analysis. Thiobarbituric acid reactive substances (TBARS) of the samples treated with 100% NaCl and HPP increased by an average of 0.52 ± 0.01 mg MDA/kg compared with the control (100% NaCl-no HPP) across the 21 storage days. In addition, 50% NaCl substitution with KCl in combinations with HPP treatments increased TBARS across the 21 storage days by an average of 0.40 ± 0.02 mg MDA/kg compared with no HPP treatment. However, on day 3, there was a sharp decrease in TBARS by an average of 0.10 ± 0.01 mg MDA/kg compared with days 0, 7, 14, and 21 in all treatments. At the end of 21 days of storage, a total of 227 volatile compounds were identified and quantified in the beef sausage, including 43 aldehydes, 46 phenols, 8 ketones, 30 alcohols, 14 acids, 12 esters, 27 terpenes, and 47 alkanes. However, no ketone compounds were detected on days 7, 14 and 21; esters on day 14 and acids on days 14 and 21 in the samples treated with or without HPP across the salts levels. However, high OAVs (OAV > 1) were obtained after partial substitution of NaCl with KCl at 25 and 50% with HPP treatment compared to the samples not treated with HPP. The aroma perceived in the beef sausage was due to compounds with the highest OAVs such as; pentadecanal, benzyl carbazate, anethole, myristicin, o-cresol, phenylacetaldehyde and (E)-methyl isoeugenol, pentadecanal, hexanoic acid, octanoic acid, eugenol, trans-2-nonenal, trans-2-octenal, trans-2-decenal, 2-butyl-1-octanol, 2,3-butanedione, ethyl hexanoate, ethyl octanoate, (-)-4-terpineol which had an OAV > 1 as compared to the other compounds with an OAV < 1. In conclusion, 25 and 50% NaCl partial replacement with KCl coupled with HPP technique can be considered in producing low-NaCl beef sausage in order to improve the flavor and decrease lipid oxidation during cold storage.

Valorization of Cattle Slaughtering Industry By-Products: Modification of the Functional Properties and Structural Characteristics of Cowhide Gelatin Induced by High Hydrostatic Pressure.

This study investigates the effects of different pressures (200, 250, 300, 350, and 400 MPa) and durations (5, 10, 15, 20, and 25 min) on the functional properties, secondary structure, and intermolecular forces of cowhide gelatin. Our results show that high hydrostatic pressure significantly affected the two, three, and four-level structures of gelatin and caused the contents of the α-helix and ß-turn to decrease by 68.86% and 78.58%, respectively (p < 0.05). In particular, the gelatin at 300 MPa for 15 min had the highest gel strength, emulsification, solubility, and foaming of all the treatment conditions under study. The analysis of the surface hydrophobicity, sulfhydryl content, zeta potential, and Raman spectroscopy shows that at a pressure of 300 MPa (15 min), the hydrogen bonds and hydrophobic interactions between collagen molecules are strongly destroyed, leading to changes in the tertiary and quaternary conformation of the protein and unfolding, with the electrostatic repulsion between protein particles making the decentralized state stable. In conclusion, moderate pressure and time can significantly improve the functional and structural properties of collagen, which provides theoretical support and guidance for realizing the high-value utilization of cowhide.

Pathological Features of Enterovirus 71-Associated Brain and Lung Damage in Mice Based on Quantitative Proteomic Analysis.

The outbreaks of enterovirus 71 (EV71)-associated hand, foot, and mouth disease (HFMD) have emerged as an emergency of global health due to its association with fatal encephalitis and subsequent neurogenic pulmonary edema; however, the molecular characteristics and pathological features underlying EV71-associated encephalitis and pulmonary edema remain largely unknown. In this study, we performed a proteomic analysis of fresh brain and lung tissues from EV71-infected mice at 7 days post infection. We detected a perturbed expression of 148 proteins in the brain and 78 proteins in the lung after EV71 expression. Further analysis showed that the dysregulated proteins in the brain are involved in a variety of fundamental biological pathways, including complement and coagulation cascades, innate and adaptive immune responses, platelet activation, and nitrogen metabolism, and those proteins in the lung participate in innate and adaptive immune responses, phagosome, arginine biosynthesis, and hypoxia-inducible factor 1 signaling pathway. Our results suggested that immune activation, complement and coagulation dysfunction, platelet activation, imbalance of nitrogen metabolism, and hypoxia could be involved in the pathogenesis of EV71, which explains the major clinical manifestation of hyperinflammatory status of severe HFMD cases. Our study provides further understanding of the molecular basis of EV71 pathogenesis.

Role of Renin-Angiotensin System in Acute Lung Injury Caused by Viral Infection.

The renin-angiotensin system (RAS) is the most important regulatory system of electrolyte homeostasis and blood pressure and acts through angiotensin-converting enzyme (ACE)/angiotensin II (Ang II)/Ang II type 1 (AT1) receptor axis and angiotensin-converting enzyme 2 (ACE2)/angiotensin (1-7)/MAS receptor axis. RAS dysfunction is related to the occurrence and development of acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) and causes a serious prognosis and even death. ALI/ARDS can be induced by various ways, one of which is viral infections, such as SARS-CoV, SARS-CoV-2, H5N1, H7N9, and EV71. This article reviews the specific mechanism on how RAS dysfunction affects ALI/ARDs caused by viral infections. SARS-CoV and SARS-CoV-2 enter the host cells by binding with ACE2. H5N1 and H7N9 avian influenza viruses reduce the ACE2 level in the body, and EV71 increases Ang II concentration. Treatment with angiotensin-converting enzyme inhibitor and angiotensin AT1 receptor blocker can alleviate ALI/ARDS symptoms. This review provides suggestions for the treatment of lung injury caused by viral infections.