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N-linked glycosylation is a common posttranslational modification of proteins that results in macroheterogeneity of the modification site. However, unlike simpler modifications, N-glycosylation introduces an additional layer of complexity with tens of thousands of possible structures arising from various dimensions, including different monosaccharide compositions, sequence structures, linking structures, isomerism, and three-dimensional conformations. This results in additional microheterogeneity of the modification site of N-glycosylation, i.e., the same N-glycosylation site can be modified with different glycans with a certain stoichiometric ratio. N-glycosylation regulates the structure and function of N-glycoproteins in a site- and structure-specific manner, and differential expression of N-glycosylation under disease conditions needs to be characterized through site- and structure-specific quantitative analysis. Numerous advanced methods ranging from sample preparation to mass spectrum analysis have been developed to distinguish N-glycan structures. Chemical derivatization of monosaccharides, online liquid chromatography separation and ion mobility spectrometry enable the physical differentiation of samples. Tandem mass spectrometry further analyzes the macro/microheterogeneity of intact N-glycopeptides through the analysis of fragment ions. Moreover, the development of search engines and AI-based software has enhanced our understanding of the dissociation patterns of intact N-glycopeptides and the clinical significance of differentially expressed intact N-glycopeptides. With the help of these modern methods, structure-specific N-glycoproteomics has become an important tool with extensive applications in the biomedical field.

Rationale: A general N‐glycoproteomics analysis pipeline has been established for characterization of mutation‐related gain‐of‐glycosylation (GoG) at intact N‐glycopeptide molecular level, generating comprehensive site and structure information of N‐glycosylation. Methods: This study focused on mutation‐originated GoG using a mass spectrometry‐based N‐glycoproteomics analysis workflow. In brief, GoG intact N‐glycopeptide databases were built, consisting of 2701 proteins (potential GoG N‐glycosites and amino acids derived from MUTAGEN, VARIANT and VAR_SEQ in UniProt) and 6709 human N‐glycans (≤50 sequence isomers per monosaccharide composition). We employed the site‐ and structure‐specific N‐glycoproteomics workflow utilizing intact N‐glycopeptides search engine GPSeeker to identify GoG intact N‐glycopeptides from parental breast cancer stem cells (MCF‐7 CSCs) and adriamycin‐resistant breast cancer stem cells (MCF‐7/ADR CSCs). Results: With the criteria of spectrum‐level false discovery rate control of ≤1%, we identified 87 and 94 GoG intact N‐glycopeptides corresponding to 37 and 35 intact N‐glycoproteins from MCF‐7 CSCs and MCF‐7/ADR CSCs, respectively. Micro‐heterogeneity and macro‐heterogeneity of N‐glycosylation from GoG intact N‐glycoproteins with VAR_SEQ and VARIANT were found in both MCF‐7 CSCs and MCF‐7/ADR CSCs systems. Conclusions: The integration of site‐ and structure‐specific N‐glycoproteomics approach, conjugating with GoG characterization, provides a universal workflow for revealing comprehensive N‐glycosite and N‐glycan structure information of GoG. The analysis of mutation‐originated GoG can be extended to GoG characterization of other N‐glycoproteome systems including complex clinical tissues and body fluids. [ABSTRACT FROM AUTHOR]

Non-imaging radar can achieve the detection and localization of moving targets, but it faces difficulties in the classification of targets. Aiming for the application of moving target classification by low/medium resolution non-imaging radar, this paper proposes a novel feature extraction method, based on the statistical characteristics of echo amplitude with the L-band fully polarimetric radar. A feature plane, composed of the 3rd-order central moment (skewness) and 4th-order central moment (kurtosis) as the statistical characteristics of the echo envelope, is established. In addition, two types of moving targets, pedestrians and non-motorized vehicles, are able to be effectively distinguished according to whether the echo signals of different polarizations have overlapping areas in the feature plane. The L-band fully polarimetric radar has been developed and field experiments have been conducted. The experimental results verify that the kurtosis and skewness of the fully polarimetric echo amplitude of the pedestrians are very close, and there is an overlapping area in the feature plane, while the statistical characteristics of the fully polarimetric echo signal of non-motor vehicles are different, and there is no overlapping area in the feature plane. This proposed feature extraction method has the advantage of being simple and robust, compared to the traditional imaging approach. [ABSTRACT FROM AUTHOR]

O-linked β- N -acetylglucosaminylation (O-GlcNAcylation) is a highly dynamic and widespread post-translational modification (PTM) that regulates the activity, subcellular localization, and stability of target proteins. O-GlcNAcylation is a reversible PTM controlled by two cycling enzymes: O-linked N -acetylglucosamine transferase and O-GlcNAcase. Emerging evidence indicates that O-GlcNAcylation plays critical roles in innate immunity, inflammatory signaling, and cancer development. O-GlcNAcylation usually occurs on serine/threonine residues, where it interacts with other PTMs, such as phosphorylation. Thus, it likely has a broad regulatory scope. This review discusses the recent research advances regarding the regulatory roles of O-GlcNAcylation in innate immunity and inflammation. A more comprehensive understanding of O-GlcNAcylation could help to optimize therapeutic strategies regarding inflammatory diseases and cancer. [ABSTRACT FROM AUTHOR]

When fluid flows through a pipe that is packed with sand particles, the fluid will bear the resistance from the sand-pack, as well as the viscous shear from the pipe wall. If the viscous shear from the pipe wall can be neglected, the fluid flow will obey Darcy's law, and one can think the equivalent permeability of the packed-pipe equals the permeability of the sand-pack. However, if the viscous shear from the pipe wall cannot be neglected, the fluid flow will obey the Brinkman equation, and the permeability of the packed-pipe will be less than that of the sand-pack due to the additional viscous drag. In this work, on the basis of the Brinkman equation, we derived a series of analytical solutions for characterizing the fluid flow in packed-pipes. These solutions can be used to depict the velocity profiles, estimate the flux rate, and calculate the equivalent permeability of a packed-pipe. On the basis of these analytical solutions, we found that Poiseuille's law is a special form of the derived equivalent permeability solution. We further divided the fluid flow in a packed-pipe into three regimes, including N-S flow, Brinkman flow, and Darcy flow. During the regime of Brinkman flow, the dimensionless flow velocity at the pipe center is 1, and the dimensionless flow velocity is gradually decreased to 0 at the pipe wall. We also investigated the effects of sorting, sand particle size, and sand-pack porosity on the packed-pipe permeability. The calculated results show that a more uniform size of the sand particles or a smaller mean particle diameter can lead to lower packed-pipe permeability. Compared to the sorting and mean particle diameter, the sand-pack porosity exerts a more significant effect on the packed-pipe permeability. [ABSTRACT FROM AUTHOR]

High-throughput proteome-level characterization of stemness markers of MCF-7 cancer stem cells was carried out using our recently developed site- and structure-specific isotopic-labeling quantitative N-glycoproteomics pipeline. 144 differentially expressed intact N-glycopeptides were quantified all at once yet with good accurate consistency with the individual biochemical measurements of proteins, RNA or DNA. [ABSTRACT FROM AUTHOR]

Histone post‐translational modifications (PTMs) play various roles in chromatin‐related cellular processes, and comprehensive analysis of these combinatorial PTMs at the intact protein level by top‐down proteomics is the method of choice to reveal their crosstalk and biological functions. Here, we report our top‐down characterization of the core histones from mouse fibroblasts cells NIH/3T3, which is a classic model used in many kinds of research. With nanoRPLC‐MS/MS analysis and ProteinGoggle database search, 547 protein species were identified with spectrum‐level FDR ≤ 1%, where PTMs in 51 protein species were unambiguously localized with PTM scores ≥1. High‐resolution MS/MS data also allowed the unambiguous identification of acetylation instead of trimethylation. This study presents a general picture of combinatorial PTMs of mouse core histones, which serves as a basic reference for all future related biological studies. [ABSTRACT FROM AUTHOR]