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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]

The separation of Co(II) and Ni(II) from leaching solution is gaining interest because Co(II) and Ni(II) are increasingly used in emerging strategic areas, such as power batteries. Herein, the surface of silica gel is functionalized with 1,2-ethylenediamine and used for the separation of Co(II) and Ni(II). The Co(II) removal efficiency of the modified silica is 80.2%, with a 4-fold improvement in the separation factor. The geometry, frequency, and electrostatic potential of the ethylenediamine modified silica gel (en/SG) are calculated. The corresponding properties of M2+ (M-Co, Ni) adsorbed on en/SG in an aqueous solution are simulated and analyzed. The results show that ethylenediamine tends to form [Men(H2O)4]2+ after binding to M2+, and the binding ability of Co(II) to ethylenediamine is stronger. Besides, the thermodynamic calculations show that en/SG has a more negative Gibbs free energy when absorbing Co(II) in aqueous solution, so en/SG is more inclined to bind with Co(II) preferentially. It is the difference in complexation ability between Ni, Co, and ethylenediamine that enlarges the difference in the original physical adsorption, thus strengthening the separation performance. This work will provide guidance for a rational design of high-performance nickel-cobalt adsorption materials. [ABSTRACT FROM AUTHOR]

Organogenesis is an important process for plant regeneration by tissue or cell mass differentiation to regenerate a complete plant. MicroRNAs (miRNAs) play an essential role in regulating plant development by mediating target genes at transcriptional and post-transcriptional levels, but the diversity of miRNAs and their potential roles in organogenesis of Acacia crassicarpa have rarely been investigated. In this study, approximately 10 million sequence reads were obtained from a small RNA library, from which 189 conserved miRNAs from 57 miRNA families, and 7 novel miRNAs from 5 families, were identified from A. crassicarpa organogenetic tissues. Target prediction for these miRNAs yielded 237 potentially unique genes, of which 207 received target Gene Ontology annotations. On the basis of a bioinformatic analysis, one novel and 13 conserved miRNAs were selected to investigate their possible roles in A. crassicarpa organogenesis by qRT-PCR. The stage-specific expression patterns of the miRNAs provided information on their possible regulatory functions, including shoot bud formation, modulated function after transfer of the culture to light, and regulatory roles during induction of organogenesis. This study is the first to investigate miRNAs associated with A. crassicarpa organogenesis. The results provide a foundation for further characterization of miRNA expression profiles and roles in the regulation of diverse physiological pathways during adventitious shoot organogenesis of A. crassicarpa. [ABSTRACT FROM AUTHOR]

Organogenesis is an important process for plant regeneration by tissue or cell mass differentiation to regenerate a complete plant. MicroRNAs (miRNAs) play an essential role in regulating plant development by mediating target genes at transcriptional and post-transcriptional levels, but the diversity of miRNAs and their potential roles in organogenesis of Acacia crassicarpa have rarely been investigated. In this study, approximately 10 million sequence reads were obtained from a small RNA library, from which 189 conserved miRNAs from 57 miRNA families, and 7 novel miRNAs from 5 families, were identified from A. crassicarpa organogenetic tissues. Target prediction for these miRNAs yielded 237 potentially unique genes, of which 207 received target Gene Ontology annotations. On the basis of a bioinformatic analysis, one novel and 13 conserved miRNAs were selected to investigate their possible roles in A. crassicarpa organogenesis by qRT-PCR. The stage-specific expression patterns of the miRNAs provided information on their possible regulatory functions, including shoot bud formation, modulated function after transfer of the culture to light, and regulatory roles during induction of organogenesis. This study is the first to investigate miRNAs associated with A. crassicarpa organogenesis. The results provide a foundation for further characterization of miRNA expression profiles and roles in the regulation of diverse physiological pathways during adventitious shoot organogenesis of A. crassicarpa. [ABSTRACT FROM AUTHOR]