Differential expression analysis of miRNAs in macrophage-derived exosomes in the tuberculosis-infected bone microenvironment

BackgroundMacrophages play an important role in regulating the course of spinal tuberculosis within the bone microenvironment. This study aimed to investigate the differential expression of miRNA in macrophage-derived exosomes within the tuberculosis-infected bone microenvironment, to identify miRNAs that hold potential as diagnostic markers and therapeutic targets.MethodsWe established study cohorts for spinal tuberculosis, collected bone marrow blood samples, isolated macrophage exosomes, and performed exosome miRNA sequencing. A miRNA-mRNA co-expression network was constructed using WGCNA analysis. Gene GO analysis and KEGG pathway enrichment analysis were performed using KOBAS software. Target miRNAs were selected based on fold change, P-value, and false discovery rate, and their validation was carried out using qRT-PCR and ROC curve studies. Subsequently, we constructed a target gene network for these miRNAs and performed KEGG pathway enrichment analysis to explore the potential signaling mechanisms involved in regulating the disease course of spinal tuberculosis.ResultsOur findings revealed that macrophages from the tuberculosis-infected bone microenvironment exhibited an M1 phenotype. The successful extraction of exosomes from macrophage supernatants was confirmed through electron microscopy, particle size analysis, and protein blot analysis. Exosome miRNA-seq demonstrated that 28 miRNAs were up-regulated, while 34 miRNAs were down-regulated in individuals with spinal tuberculosis. GO analysis and KEGG pathway enrichment analysis indicated that the differentially expressed miRNAs were involved in various biological processes, cell components, molecular functions, and signaling pathways, which collectively contribute to the regulation of the disease course of spinal tuberculosis. Notably, miRNA-125b-5p was successfully selected based on fold change, p-value, and false discovery rate. qRT-PCR validation further confirmed the significant up-regulation of miRNA-125b-5p in spinal tuberculosis. The ROC curve revealed that miR-125b-5p is a potential diagnostic biomarker for spinal tuberculosis. Moreover, construction of the miRNA-125b-5p target gene network and subsequent KEGG enrichment analysis highlighted the importance of MAPK, TNF, Ras, Rap1, and the PI3K-Akt signaling pathways in the regulation of the disease course of spinal tuberculosis.ConclusionOur study demonstrates differential expression of miRNAs in macrophage-derived exosomes in the tuberculosis-infected bone microenvironment. Specifically, MiRNA-125b-5p is significantly up-regulated in spinal tuberculosis and shows potential as a diagnostic biomarker for spinal tuberculosis

Changes in glycosylated proteins in colostrum and mature milk and their implication

IntroductionGlycosylation is one of the essential post-translational modifications that influences the function of milk proteins.MethodsIn the present study, 998 proteins and 764 glycosylated sites from 402 glycoproteins were identified in human milk by TMT labeling proteomics. Compared to human milk proteins, the glycoproteins were mainly enriched in cell adhesion, proteolysis, and defense/immune process.ResultsThe abundance of 353 glycosylated sites and their 179 parent proteins was quantified. After normalization to their parent protein’s abundance, 78 glycosylated sites in 56 glycoproteins and 10 glycosylated sites in 10 glycoproteins were significantly higher in colostrum and mature milk, respectively. These changed glycoproteins were mainly related to host defense. Intriguingly, one glycosylated site (Asp144) in IgA and two glycosylated sites (Asp38 and Asp1079) in tenascin are significantly upregulated even though their protein abundance was downregulated during lactation.DiscussionThis study helps us figure out the critical glycosylated sites in proteins that might influence their biological function in an unbiased way

Fatty Acid, Triglyceride, and Kinetic Properties of Milk Fat Fractions Made by the Combination of Dry Fractionation and Short-Path Molecular Distillation.

peer reviewedIn this study, we aimed to detect the physicochemical properties of distilled products (residue and distillate) obtained from anhydrous milk fat (AMF) and its dry fractionation products (liquid and solid fractions at 25°C (25 L and 25 S)). The results showed that the saturated fatty acids and low- and medium-molecular-weight triglycerides were easily accumulated in the distillate, and the percentage of unsaturated fatty acid and high-molecular-weight triglycerides in the residue were higher, and these components in 25 S and 25 L were influenced more significantly than those in the AMF. In addition, the distillate had larger melting ranges in comparison with the distilled substrate, while the melting ranges of residue was smaller. The triglycerides were presented as the mixture crystal forms (α, β', and β crystal) in 25 S, AMF, and their distilling products, and it was transformed gradually to a single form as the increasing of distilling temperature. Moreover, the accumulated pattern of triglycerides was double chain length in 25 S, AMF, and their distilling products. These results provide a new approach to obtain the MF fractions with different properties, and the findings of this study enrich the theoretical basis of MF separation in practical production

Analysis of the non-volatile components and volatile compounds of hydrolysates derived from unmatured cheese curd hydrolysis by different enzymes

peer reviewedFifteen cheese protein hydrolysates were produced by using four different proteases. Then, the free amino acids (FAAs), molecular weight distribution (MWD), electronic tongue evaluation, and 45 volatile compounds of the corresponding products were evaluated, respectively. The results 46 suggested that 2SD had the strongest hydrolysis characteristic, followed by 6SD and FN. Samples 47 hydrolyzed for less than 6 h or more than 18 h contained great defects of taste. Peptides with 150 48 Da – 450 Da were mainly responsible for bitterness, saltiness, umami, and aftertaste in some enzyme 49 hydrolysis. Under the same total enzyme concentration condition, the sample hydrolyzed by 50 Flavourzyme and Neutrase for 18 h released more richness and less bitterness than the other systems, 51 which were characterized by butter and cream odor. Notably, it was found for the first time that 52 tetramethylpyrazine (TMP) was detected in cheese proteolysis with the highest content of 17.59 53 µg/g in Protease 2SD for 30 h. 2-Undecanone and acetoin played a key role in the flavor formation 54 of the tested samples. Regarding the different chemical families of volatiles, acids were more 55 abundant in the samples hydrolyzed by Protease 2SD and 6SD, while FN systems can achieve high 56 ketone content

Free fatty acid hydrolyzed with lipases and their effects on enzyme-modified cheese flavor

peer reviewed: This study investigated the effects of five lipases on enzyme-modified cheese (EMC) flavor development. Results showed that lipase 30SD contained high hydrolytic activity for short, medium, and long-chain fatty acids within 24 h incubation time, and the highest content of them among different times could reach 47.24, 475.90, 1 563.92 mg/100 g fat, respectively. Lipase DF15 and MER showed moderate capacity to hydrolyze volatile fatty acids, while lipase F3G had a stronger ability to produce long-chain fatty acids. Twenty-seven new volatiles were formed during lipolysis, most of them were acids and esters. Principal component analysis results showed that EMC produced by lipase 30SD for 18 h was similar to the commercial product with a pungent, rancid, and cheddar flavor. EMCs produced by lipase DF15 were significantly distinguished from other products by their high content of ethyl heptanoate, ethyl nonanoate, and ethyl tridecanoate. The findings might be useful for the researchers who focus on lipolysis or EMC product

Identification and Functional Validation of Autolysis—Associated Genes in Lactobacillus bulgaricus ATCC BAA-365

Lactic acid bacteria (LAB) are important organisms in food production. Indeed, LAB autolysis is very critical in dairy processing. For example, it influences the development of cheese flavor by releasing intracellular enzymes, and controls cell growth in yogurts and probiotic products. Two component systems (TCS) constitute essential environmental sensors and effectors of signal transduction in most bacteria. In the present work, mutants of one TCS (LBUL_RS00115/LBUL_RS00110) were generated to assess the relationship between TCS and cell autolysis. The mutants displayed decreased autolysis in comparison with wild type; meanwhile, complementation reversed this effect. The interaction between LBUL_RS00115 and LBUL_RS00110 was confirmed by yeast two-hybrid analysis. These observations suggested that the TCS (LBUL_RS00115/LBUL_RS00110) was involved in autolysis in Lactobacillus delbrueckii subsp. bulgaricus

Advances on chimeric antigen receptor-modified T-cell therapy for oncotherapy

Abstract Tumor treatment is still complicated in the field of medicine. Tumor immunotherapy has been the most interesting research field in cancer therapy. Application of chimeric antigen receptor T (CAR-T) cell therapy has recently achieved excellent clinical outcome in patients, especially those with CD19-positive hematologic malignancies. This phenomenon has induced intense interest to develop CAR-T cell therapy for cancer, especially for solid tumors. However, the performance of CAR-T cell treatment in solid tumor is not as satisfactory as that in hematologic disease. Clinical studies on some neoplasms, such as glioblastoma, ovarian cancer, and cholangiocarcinoma, have achieved desirable outcome. This review describes the history and evolution of CAR-T, generalizes the structure and preparation of CAR-T, and summarizes the latest advances on CAR-T cell therapy in different tumor types. The last section presents the current challenges and prospects of CAR-T application to provide guidance for subsequent research

Novel SGH Recognition Algorithm Based Robot Binocular Vision System for Sorting Process

To achieve automatic sorting on commodity trademarks, a binocular vision system has been constructed in this paper. By adjusting camera pose, this system can obtain greater shooting perspective. In order to improve sorting accuracy, a now SGH recognition method is proposed. SGH consists of spatial color histogram (S feature), gray level cooccurrence matrix (G feature), and Hu moments (H) feature, which represent color feature, texture feature, and shaper feature, respectively. Similarity judgment function is built by using SGH. The experimental results show that SGH algorithm has a higher visual accuracy compared to single feature based recognition method

Screening for Cholesterol-Lowering Probiotics from Lactic Acid Bacteria Isolated from Corn Silage Based on Three Hypothesized Pathways

A total of 85 strains of lactic acid bacteria were isolated from corn silage in this study and analyzed in vitro for their cholesterol removal, NPC1L1 protein down-regulation and bile salt deconjugation ability, respectively. Nineteen strains were selected for further analysis for their probiotic potential. Finally, 3 strains showing better probiotic potential were evaluated for their cholesterol-lowering activity in hamsters. The strains showing the greater cholesterol removal and NPC1L1 protein down-regulation activity had no significant effects on serum and hepatic cholesterol levels in hamsters (p > 0.05). However, Lactobacillus plantarum CAAS 18008 (1 × 109 CFU/d) showing the greater bile salt deconjugation ability significantly reduced serum low-density lipoprotein cholesterol, total cholesterol, and hepatic total cholesterol levels by 28.8%, 21.7%, and 30.9%, respectively (p < 0.05). The cholesterol-lowering mechanism was attributed to its bile salt hydrolase activity, which enhanced daily fecal bile acid excretion levels and thereby accelerated new bile acid synthesis from cholesterol in liver. This study demonstrated that the strains showing greater cholesterol removal and NPC1L1 protein down-regulation activity in vitro hardly reveal cholesterol-lowering activity in vivo, whereas the strains showing greater bile salt deconjugation ability in vitro has large potential to decrease serum cholesterol levels in vivo