Large-scale identification of N-linked intact glycopeptides by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) in human serum is challenging because of the wide dynamic range of serum protein abundances, the lack of a complete serum N-glycan database and the existence of proteoforms. In this regard, a spectral library search method was presented for the identification of N-linked intact glycopeptides from N-linked glycoproteins in human serum with target-decoy and motif-specific false discovery rate (FDR) control. Serum proteins were firstly separated into low-abundance and high-abundance proteins by acetonitrile (ACN) precipitation. After digestion, the N-linked intact glycopeptides were enriched by hydrophilic interaction liquid chromatography (HILIC) and a portion of the enriched N-linked intact glycopeptides were processed by Peptide-N-Glycosidase F (PNGase F) to generate N-linked deglycopeptides. Both N-linked intact glycopeptides and deglycopeptides were analyzed by LC-MS/MS. From N-linked deglycopeptides data sets, 764 N-linked glycoproteins, 1699 N-linked glycosites and 3328 unique N-linked deglycopeptides were identified. Four types of N-linked glycosylation motifs (NXS/T/C/V, X≠P) were used to recognize the N-linked deglycopeptides. The spectra of these N-linked deglycopeptides were utilized for N-linked deglycopeptides library construction and identification of N-linked intact glycopeptides. A database containing 739 N-glycan masses was constructed and utilized during spectral library search for the identification of N-linked intact glycopeptides. In total, 526 N-linked glycoproteins, 1036 N-linked glycosites, 22,677 N-linked intact glycopeptides and 738 N-glycan masses were identified under 1% FDR, representing the most in-depth serum N-glycoproteome identified by LC-MS/MS at N-linked intact glycopeptide level.

Confident characterization of intact glycopeptides is a challenging task in mass spectrometry-based glycoproteomics due to microheterogeneity of glycosylation, complexity of glycans, and insufficient fragmentation of peptide bones. Open mass spectral library search is a promising computational approach to peptide identification, but its potential in the identification of glycopeptides has not been fully explored. Here we present pMatchGlyco, a new spectral library search tool for intact N-linked glycopeptide identification using high-energy collisional dissociation (HCD) tandem mass spectrometry (MS/MS) data. In pMatchGlyco, (1) MS/MS spectra of deglycopeptides are used to create spectral library, (2) MS/MS spectra of glycopeptides are matched to the spectra in library in an open (precursor tolerant) manner and the glycans are inferred, and (3) a false discovery rate is estimated for top-scored matches above a threshold. The efficiency and reliability of pMatchGlyco were demonstrated on a data set of mixture sample of six standard glycoproteins and a complex glycoprotein data set generated from human cancer cell line OVCAR3.

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In nature, geckos have extraordinary adhesive capabilities. The multi-scale hierarchical structure of the gecko foot hairs, especially the high-aspect-ratio structure of its micro-scale seta and nano-scale spatulae is the critical factor of the gecko's ability to adopt and stick to any different surface with powerful adhesion force. In this paper, we present a simple and effective approach to fabricate dual-level hierarchical structures mimicking gecko foot hairs. Polydimethyl-siloxane (PDMS) hierarchical arrays were fabricated by demolding from a double stack mold that was composed of an SU-8 mold by thick film photolithography and a silicon mold by inductively coupled plasma (ICP) etching. Top pillars of the fabricated structures have 3 micom diameter and 18 microm in height, while base pillars have 25 microm diameter and 40 microm in height. The water droplet contact angle tests indicate that the hierarchical structures increase the hydrophobic property significantly compared with the single-level arrays and the unstructured polymers, exhibiting superhydrophobicity (154.2 degrees) like the Tokay gecko's (160.9 degrees). The shear force tests show that the top pillars make attachment through side contact with a value of about 0.25 N/cm2, and moreover, the hierarchical structures are demonstrated to be more suitable for contacting with rough surfaces.

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Solar radiation intensity is intermittent and uncertain under the influence of meteorological conditions. Clustering them and obtaining high-precision and reliable probabilistic forecasting results play a vital role in the planning and management of solar power. In this study, a novel K-means time series clustering (K-MTSC) algorithm is first proposed to cluster solar radiation intensity and compared with astronomy method and K-means. Then, different feature inputs for different categories of solar radiation intensity are screened. Afterwards, the different kernel functions of Gaussian process regression (GPR) are compared and optimal kernel function is selected in terms of deterministic forecasting and probabilistic forecasting for different categories. Finally, the case study in Tibet province, China are performed to verify the validity and practicability of this research model and method. In this experiment, the average accuracy of GPR is 44% higher than that of Artificial Neural Network ANN, and 17% higher than that of Support Vector Regression. The experiments show that (1) the clustering results obtained by the K-MTSC algorithm have a larger inter-group distance and a smaller intra-group distance, and at the same time, it will not destroy the continuity of the time series. (2) The probability forecast results obtained by GPR are reliable and high-accuracy.INDEX TERMS solar radiation intensity; K-means Time Series Clustering; probabilistic forecasting; Gaussian process regression.

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