Bovine milk-derived exosomes (BMDEs) have potential applications in the pharmaceutical industry as drug delivery carriers. A comprehensive analysis of protein glycosylation in exosomes is necessary to elucidate the process of targeted delivery. In this work, free oligosaccharides (FOSs), O-glycans, and N-glycans in BMDEs and whey were first analyzed through multiple derivation strategies. In summary, 13 FOSs, 44 O-glycans, and 94 N-glycans were identified in bovine milk. To analyze site-specific glycosylation of glycoproteins, a one-step method was used to enrich and characterize intact glycopeptides. A total of 1359 proteins including 114 glycoproteins were identified and most of these were located in the exosomes. Approximately 95 glycopeptides were initially discovered and 5 predicted glycosites were confirmed in BMDEs. Collectively, these findings revealed the characterization and distribution of glycans and glycoproteins in BMDEs, providing insight into the potential applications of BMDEs in drug delivery and food science.

STT3A and STT3B are the main catalytic subunits of the oligosaccharyltransferase complex (OST-A and OST-B in mammalian cells), which primarily mediate cotranslational and post-translocational N-linked glycosylation, respectively. To determine the specificity of STT3A and STT3B, we performed proteomic and glycoproteomic analyses in the gene knock-out (KO) and wild-type HEK293 cells. In total, 3961 proteins, 4265 unique N-linked intact glycopeptides and 629 glycosites representing 349 glycoproteins were identified from all these cells. Deletion of the STT3A gene had a greater impact on the protein expression than deletion of STT3B, especially on glycoproteins. In addition, total mannosylated N-glycans were reduced and fucosylated N-glycans were increased in STT3A-KO cells, which were caused by the differential expression of glycan-related enzymes. Interestingly, hyperglycosylated proteins were identified in KO cells, and the hyperglycosylation of ENPL was caused by the endoplasmic reticulum (ER) stress due to the STT3A deletion. Furthermore, the increased expression of the ATF6 and PERK indicated that the unfolded protein response also happened in STT3A-KO cells. Overall, the specificity of STT3A and STT3B revealed that defects in the OST subunit not only broadly affect N-linked glycosylation of the protein but also affect protein expression.

Schistosomiasis is a chronic parasitic disease that continues to be a pressing public health problem in many developing countries. The primary pathological damage from the disease is granuloma and fibrosis caused by egg aggregation, and early treatment can effectively prevent the occurrence of liver fibrosis. Therefore, it is very important to identify biomarkers that can be used for early diagnosis of Schistosoma japonicum infection. In this study, a label-free proteomics method was performed to observe the alteration of proteins before infection, 1 and 6 weeks after infection, and 5 and 7 weeks after treatment. A total of 10 proteins derived from S. japonicum and 242 host-derived proteins were identified and quantified as significantly changed. Temporal analysis was carried out to further analyze potential biomarkers with coherent changes during infection and treatment. The results revealed biological process changes in serum proteins compared to infection and treatment groups, which implicated receptor-mediated endocytosis, inflammatory response, and acute-phase response such as mannan-binding lectin serine peptidase 1, immunoglobulin, and collagen. These findings offer guidance for the in-depth analysis of potential biomarkers of schistosomiasis, host protein, and early diagnosis of S. japonicum and its pathogenesis. Data are available via ProteomeXchange with identifier PXD029635.

To solve the problem of parameter selection during the design of magnetically coupled resonant wireless power transmission system (MCR-WPT), this paper proposed an improved genetic simulated annealing algorithm. Firstly, the equivalent circuit of the system is analysis in this study and a nonlinear programming mathematical model is built. Secondly, in place of the penalty function method in the genetic algorithm, the selection strategy based on the distance between individuals is adopted to select individual. In this way, it reduces the excess empirical parameters. Meanwhile, it can improve the convergence rate and the searching ability by calculating crossover probability and mutation probability according to the variance of population’s fitness. At last, the simulated annealing operator is added to increase local search ability of the method. The simulation shows that the improved method can break the limit of the local optimum solution and get the global optimum solution faster. The optimized system can achieve the practical requirements.

Background Breast cancer is one of the most frequently occurring malignant cancers worldwide. Invasive ductal carcinoma (IDC) and invasive lobular carcinoma (ILC) are the two most common histological subtypes of breast cancer. The prognosis and survival of the two subtypes were significantly different even with specific molecular subgroup. In this study, we aimed to deeply explore molecular characteristics and the relationship between IDC and ILC subtypes in same molecular subgroup of breast cancer using comprehensive proteomics and phosphoproteomics analysis. Methods Cancer tissues and noncancerous adjacent tissues (NATs) with the luminal subtype (ER- and PR-positive, HER2-negative) were obtained from paired IDC and ILC patients respectively. Label-free quantitative proteomics and phosphoproteomics methods were used to detect differentially expressed proteins and the phosphorylation status between 10 paired breast cancer and NATs. Then, bioinformatics analysis was performed to explore the difference between IDC and ILC subtypes, including the difference in protein expression levels and the degree of phosphorylation. Meanwhile, Kinase-Substrate Enrichment Analysis (KSEA) revealed the activation difference of kinases and their substrates between IDC and ILC. Results A total of 5,044 high-confidence proteins and 3,808 phosphoproteins were identified from breast cancer tissues. The protein phosphorylation level in ILC tissues was higher than that in IDC tissues. From the quantitative analysis of 1259 proteins and 560 phosphoproteins with high patient coverage, Histone H1.10, Complement C4-B and Crk-like protein were found to be significantly differentially expressed in the two subtype tissues from the proteomics analysis. Moreover, the differences in protein expression of Septin-2, Septin-9, Heterogeneous nuclear ribonucleoprotein A1 and Kinectin and their phosphorylation clearly distinguished IDC from ILC. In addition, differentially expressed proteins and differentially expressed phosphoproteins in IDC were primarily related to energy metabolism and MAPK pathway, while ILC subtypes were more closely involved in the AMPK and p53/p21 pathways. Furthermore, the kinomes from IDC were primarily significantly activated in the CMGC (Cyclin-dependent kinases, Mitogen-activated protein kinases, Glycogen synthase kinases and CDK-like kinases) groups. Conclusions Our research provides insights into the molecular characterization of IDC and ILC and contributes to discovering novel targets for further drug development and targeted treatment.