Amount Of Sample Research Articles

Enhanced Proteomic Coverage in Tissue Microenvironment by Immune Cell Subtype Library-Assisted DIA-MS

Immune cells that infiltrate the tumor microenvironment (TME) play crucial roles in shaping cancer development and influencing clinical outcomes and therapeutic responses. However, obtaining a comprehensive proteomic snapshot of tumor-infiltrating immunity in clinical specimens is often hindered by small sample amounts and a low proportion of immune infiltrating cells in the TME. To enable in-depth and highly sensitive profiling of microscale tissues, we established an immune cell-enriched library-assisted strategy for data-independent acquisition mass spectrometry (DIA-MS). Firstly, six immune cell subtype-specific spectral libraries were established from sorted cluster of differentiation markers, CD8+, CD4+ T lymphocytes, B lymphocytes, natural killer cells, dendritic cells, and macrophages in murine mesenteric lymph nodes (MLNs), covering 7815 protein groups with surface markers and immune cell-enriched proteins. The feasibility of microscale immune proteomic profiling was demonstrated on 1 μg tissue protein from the tumor of murine colorectal cancer (CRC) models using single-shot DIA; the immune cell-enriched library increased coverage to quantify 7419 proteins compared to directDIA analysis (6978 proteins). The enhancement enabled the mapping of 841 immune function-related proteins and exclusive identification of many low-abundance immune proteins, such as CD1D1, and CD244, demonstrating high sensitivity for immune landscape profiling. This approach was used to characterize the MLNs in CRC models, aiming to elucidate the mechanism underlying their involvement in cancer development within the TME. Even with a low percentage of immune cell infiltration (0.25–3%) in the tumor, our results illuminate downregulation in the adaptive immune signaling pathways (such as C-type lectin receptor signaling, and chemokine signaling), T cell receptor signaling, and Th1/Th2/Th17 cell differentiation, suggesting an immunosuppressive status in MLNs of CRC model. The DIA approach using the immune cell-enriched libraries showcased deep coverage and high sensitivity that can facilitate illumination of the immune proteomic landscape for microscale samples.

Lectin-Based SP3 Technology Enables N-Glycoproteomic Analysis of Mouse Oocytes.

N-glycosylation is one of the most universal and complex protein post-translational modifications (PTMs), and it is involved in many physiological and pathological activities. Owing to the low abundance of N-glycoproteins, enrichment of N-glycopeptides for mass spectrometry analysis usually requires a large amount of peptides. Additionally, oocyte protein N-glycosylation has not been systemically characterized due to the limited sample amount. Here, we developed a glycosylation enrichment method based on lectin and a single-pot, solid-phase-enhanced sample preparation (SP3) technology, termed lectin-based SP3 technology (LectinSP3). LectinSP3 immobilized lectin on the SP3 beads for N-glycopeptide enrichment. It could identify over 1100 N-glycosylation sites and 600 N-glycoproteins from 10 μg of mouse testis peptides. Furthermore, using the LectinSP3 method, we characterized the N-glycoproteome of 1000 mouse oocytes in three replicates and identified a total of 363 N-glycosylation sites from 215 N-glycoproteins. Bioinformatics analysis revealed that these oocyte N-glycoproteins were mainly enriched in cell adhesion, fertilization, and sperm-egg recognition. Overall, the LectinSP3 method has all procedures performed in one tube, using magnetic beads. It is suitable for analysis of a low amount of samples and is expected to be easily adaptable for automation. In addition, our mouse oocyte protein N-glycosylation profiling could help further characterize the regulation of oocyte functions.

Effect of heating processes on bioactive properties, phenolic components and mineral amounts of rocket (Eruca sativa Mill.) leaves

SummaryTotal carotenoid amounts of rocket samples were established between 120.56 μg g−1 (fresh) and 130.06 μg g−1 (microwave). In addition, total phenol and flavonoid results of rockets dried in microwave and oven were found between 655.53 (oven) and 1790.96 mg GAE/100 g (fresh) to 254.03 (oven) and 1668.83 mg/100 g (fresh), respectively. Also, radical scavenging activities (DPPH and FRAP) of rocket samples were measured between 7.55 (oven) and 25.45 mmol kg−1 (fresh) to 35.59 (oven) and 66.74 mg g−1 (fresh), respectively. Gallic acid, 3,4‐dihydroxybenzoic acid, catechin and rutin were the main phenolic compounds found in rocket samples. Catechin contents of rockets dried in microwave and oven systems were recorded between 349.60 (oven) and 442.47 mg/100 g (fresh). Gallic acid and 3,4‐dihydroxybenzoic acid amounts of rocket samples were monitored between 52.84 (oven) and 952.15 mg/100 g (fresh) to 30.92 (oven) and 282.15 mg/100 g (fresh), respectively. The protein amounts of rockets were determined between 11.00% (microwave) and 11.60% (oven). P and K values of fresh and dried rockets were found between 3148.36 (oven) and 3752.33 mg kg−1 (fresh) to 39 363.75 (microwave) and 36 883.41 mg/kg (fw), respectively. Also, Ca and Mg results of fresh and dried‐rockets were found between 26 739.33 (microwave) and 29 534.48 mg kg−1 (fresh) to 3125.32 (microwave) and 3431.70 mg kg−1 (fresh), respectively. The highest S (14 665.30 mg kg−1) was detected in rocket sample dried in microwave.

Organometallic Chirality Sensing via "Click"-Like η6-Arene Coordination with an Achiral Cp*Ru(II) Piano Stool Complex.

Piano stool complexes have been studied over many years and found widespread applications in organic synthesis, catalysis, materials and drug development. We now report the first examples of quantitative chiroptical molecular recognition of chiral compounds through click-like η6-arene coordination with readily available half sandwich complexes. This conceptually new approach to chirality sensing is based on irreversible acetonitrile displacement of [Cp*Ru(CH3CN)3]PF6 by an aromatic target molecule, a process that is fast and complete within a few minutes at room temperature. The metal coordination coincides with characteristic circular dichroism inductions that can be easily correlated to the absolute configuration and enantiomeric ratio of the bound molecule. A relay assay that decouples the determination of the enantiomeric composition and of the total sample amount by a practical CD/UV measurement protocol was developed and successfully tested. The introduction of piano stool complexes to the chiroptical sensing realm is mechanistically unique and extends the scope of currently known methods with small-molecule probes that require the presence of amino, alcohol, carboxylate or other privileged functional groups for binding of the target compound. A broad application range including pharmaceutically relevant multifunctional molecules and the use in chromatography-free asymmetric reaction analysis are also demonstrated.

Organometallic Chirality Sensing via “Click”‐Like η6‐Arene Coordination with an Achiral Cp*Ru(II) Piano Stool Complex

AbstractPiano stool complexes have been studied over many years and found widespread applications in organic synthesis, catalysis, materials and drug development. We now report the first examples of quantitative chiroptical molecular recognition of chiral compounds through click‐like η6‐arene coordination with readily available half sandwich complexes. This conceptually new approach to chirality sensing is based on irreversible acetonitrile displacement of [Cp*Ru(CH3CN)3]PF6 by an aromatic target molecule, a process that is fast and complete within a few minutes at room temperature. The metal coordination coincides with characteristic circular dichroism inductions that can be easily correlated to the absolute configuration and enantiomeric ratio of the bound molecule. A relay assay that decouples the determination of the enantiomeric composition and of the total sample amount by a practical CD/UV measurement protocol was developed and successfully tested. The introduction of piano stool complexes to the chiroptical sensing realm is mechanistically unique and extends the scope of currently known methods with small‐molecule probes that require the presence of amino, alcohol, carboxylate or other privileged functional groups for binding of the target compound. A broad application range including pharmaceutically relevant multifunctional molecules and the use in chromatography‐free asymmetric reaction analysis are also demonstrated.

Microwave drying of black rosehip (Rosa pimpinellifolia L.) fruit: optimization for enhanced anthocyanin content and analysis of physicochemical properties and bioaccessibility

Black rosehip (Rosa pimpinellifolia L.) fruit is a rich source of anthocyanins and has been promoted for several bioactive properties. Drying this fruit can increase its shelf life and enhance its applications in functional food products. This study aimed to optimize the microwave drying process of black rosehip fruit. The process was optimized based on the effect of microwave power (300, 500, and 650 W) and the amount of sample per batch (50, 75, and 100 g) on the amount of total anthocyanins and individual anthocyanins (cyanin chloride, cyanidin-3-glucoside, cyanidin-3-rutinoside, pelargonidin-3-glucoside, and cyanidin chloride). The effect of different drying conditions on color, microstructure, and texture was also investigated. The bioaccessibility of anthocyanins from the dried food sample at optimum conditions was assessed by in vitro gastrointestinal digestion. The results showed that microwave power and the amount of sample had a significant effect on the anthocyanin content of black rosehip fruit. The optimum conditions were determined to be 350 W power and 50 g sample mass. Lower microwave power and less sample mass produced dried fruits which were slightly darker, less red and less yellow. The hardness of dried fruit was also significantly affected by the microwave drying conditions. The dried black rosehip at optimum conditions showed a high bioaccessibility of cyanidin-3-glucoside (82.6 ±4.1%) which is known for a wide range of bioactive properties.

Micro-sample Nd isotopic measurement on reference rock powders via the thermal ionization mass spectrometer Nd+ mode

The Sm–Nd isotopic system has been broadly utilized for dating and tracing in earth and planetary sciences. However, low Nd+ ionization efficiency permanently hinders precise and accurate measurements on small amounts of Nd samples. So far, isotopic measurement of micro-amount Nd sample via thermal ionization mass spectrometry (TIMS) Nd+ mode was mostly performed on Nd reference solutions, but less on natural rock or mineral samples. In the present study, analytical procedures to measure isotopic composition via the TIMS Nd+ mode were designed for natural rock powders by using 1–5 ng pure Nd. The analytical technique was refined through low-blank sample digestion, small-column chemistry for Nd extraction from matrix elements, loading Nd using purified H3PO4 solution, and careful heating of filaments before data acquisition. Measurements on small Nd amounts (1 ng–5 ng) of either the JNdi-1 reference solution or four reference rock powders yielded satisfactory results of 143Nd/144Nd value with respect to precision and accuracy. The analytical results agree well with the recommendation values of the reference materials within analytical errors and the internal precision is better than ±0.005 % (2RSE). When using 1 ng or 2 ng Nd amounts, 144Nd ion flow can remain stable in a signal intensity of >100 mV, and without the assistance of an additional ionization enhancer. Therefore, the analytical procedure, featured by low procedural blank (about 10–22 pg), high Nd+ ionization efficiency, and easy operation, has broad prospects for the application of Nd isotope in single-grain minerals or micro-samples.

A multiplex digital PCR assay for detection and quantitation of porcine circovirus type 2 and type 3.

Porcine circovirus (PCV) has become a major pathogen, causing major economic losses in the global pig industry, and PCV type 2 (PCV2) and 3 (PCV3) are distributed worldwide.We designed specific primer and probe sequences targeting PCV2 Cap and PCV3 Rap anddeveloped a multiplex crystal digital PCR (cdPCR) method after optimizing the primerconcentration, probe concentration, and annealing temperature.The multiplex cdPCR assay permits precise and differential detection of PCV2 and PCV3, with a limit of detection of 1.39 × 101 and 1.27 × 101 copies/reaction, respectively, and no cross-reaction with other porcine viruses was observed. The intra-assay and interassay coefficients of variation (CVs) were less than 8.75%, indicating good repeatability and reproducibility. To evaluate the practical value of this assay, 40 tissue samples and 70 feed samples were tested for both PCV2 and PCV3 by cdPCR and quantitative PCR (qPCR). Using multiplex cdPCR, the rates of PCV2 infection, PCV3 infection, and coinfection were 28.45%, 1.72%, and 12.93%, respectively, and using multiplex qPCR, they were 25.00%, 0.86%, and 4.31%, respectively This highly specific and sensitive multiplex cdPCR thus allows accurate simultaneous detection of PCV2 and PCV3, and it is particularly well suited for applications that require the detection of small amounts of input nucleic acid or samples with intensive processing and complex matrices.

Rapid in-situ calibration of computational micro-spectrometer with few-shot meta-learning

Computational micro-spectrometers comprised of detector arrays and encoding structure arrays, such as on-chip Fabry-Perot (FP) cavity filters, have great potential in many in-situ applications owing to their compact size and snapshot imaging ability. Given manufacturing deviation and environmental influence are inevitable, easy and effective calibration for spectrometer is necessary, especially for in-situ applications. Currently calibration strategies based on iterative algorithms or neural networks require accurate measurements of pixel-level (spectral) encoding functions through monochromator or large amounts of standard samples. These procedures are time-consuming and expensive, thereby impeding in-situ applications. Meta-learning algorithms with few-shot learning ability can address this challenge by incorporating the prior knowledge in the simulated dataset. In this work, we propose a meta-learning algorithm free of measuring encoding function or large amounts of standard samples to calibrate a micro-spectrometer with manufacturing deviation effectively. Our micro-spectrometer comprises 16 types of FP filters covering a wavelength range of 550-720 nm. The center wavelength of each filter type deviates from the design up to 6 nm. After calibration with 15 different color data, the average reconstruction error on the test dataset decreased from 7.2 × 10 − 3 to 1.2 × 10 − 3, and further decreased to 9.4 × 10 − 4 when the calibration data increased to 24. The performance is comparable to algorithms trained with measured encoding function both in reconstruction error and generalization ability. We estimated that the cost of in-situ calibration through reflectance measurements of color chart decreased to one percent of the cost through monochromator measurements. By exploiting prior deviation information in simulation data with meta-learning, the efficiency and cost of calibration are significantly improved, thereby facilitating the large-scale production and in-situ application of micro-spectrometers.

Choline chloride-L-lactic acid mixtures as solvents for extraction of Coumarin from cinnamon-containing foods

The determination of coumarin in generally available cinnamon-containing foods such as biscuits, cereals is important to ensure that the populations consuming these foods more frequently are not exposed to higher levels and are not at risk. This study describes the development of an analytical method for determination of coumarin in foods containing cinnamon. Solid-liquid phase extraction with conventional stirring-assisted and ultrasound-assisted extraction (UAE) procedures were used to prepare sample extracts for liquid chromatography analysis. The green solvent, a mixture of choline chloride:L-lactic acid (1:4, mol/mol)/water (50:50 v/v), as a deep eutectic solvent-like mixture, was selected as an extraction solvent. The extraction conditions were optimized by using the design of the experiment strategy obtaining parameters, temperature 47 °C, time 36 min, solvent volume 4 mL, sample amount 0.8 g, stirring speed 20 rpm for rotary stirring-assisted extraction procedure, and temperature 30 °C, time 5 min, solvent volume 3.5 mL, sample amount 0.8 g for UAE. High-performance liquid chromatography separation with a total analysis time of 12 min was performed with a stationary phase of octadecyl type and a gradient of mobile phase of methanol-1 % acetic acid. The methods were successfully validated to show the detection limit of 0.13 μg.mL−1, the linearity range of 0.4–50 µg.mL−1 (R2 = 0.999), recovery values more than 80 %, and the RSDs of less than 7 %. Rotary stirring-assisted extraction showed moderately higher recovery values than UAE, while lower RSD % was observed for UAE.

Weakly Supervised Learning Method for Semantic Segmentation of Large-Scale 3D Point Cloud Based on Transformers

Abstract. Nowadays, semantic segmentation results of 3D point cloud have been widely applied in the fields of robotics, autonomous driving, and augmented reality etc. Thanks to the development of relevant deep learning models (such as PointNet), supervised training methods have become hotspot, in which two common limitations exists: inferior feature representation of 3D points and massive annotations. To improve 3D point feature, inspired by the idea of transformer, we employ a so-call LCP network that extracts better feature by investigating attentions between target 3D points and its corresponding local neighbors via local context propagation. Training transformer-based network needs amount of training samples, which itself is a labor-intensive, costly and error-prone work, therefore, this work proposes a weakly supervised framework, in particular, pseudo-labels are estimated based on the feature distances between unlabeled points and prototypes, which are calculated based on labeled data. The extensive experimental results show that, the proposed PL-LCP can yield considerable results (67.6% mIOU for indoor and 67.3% for outdoor) even if only using 1% real labels, and comparing to several state-of-the-art method using all labels, we achieve superior results in mIOU, OA for indoor (65.9%, 89.2%).

Individual identification method using samples associated with doping tests: A comparison of mitochondrial and nuclear genetic data.

Doping offenses involve the use or attempted use of any prohibited method or substance as well as substituting samples. Consequently, it has been recommended that short tandem repeat (STR) analysis be used to determine if the doping control samples are from the same athlete. However, it has been recognized that it may be difficult to obtain full STR analysis using negligible amounts of DNA samples. Mitochondrial DNA (mtDNA) is characterized by its stability and high cellular copy number. Therefore, mtDNA testing in urine is expected to be used to analyze samples that cannot be analyzed using STR analysis. The objective of this study was to compare mtDNA testing with STR analysis by conducting sensitivity, concordance (whole blood, dried blood spot, and urine), and case-type studies. In sensitivity studies, mtDNA testing exhibited greater sensitivity compared with STR analysis. Concordance studies indicated that all samples were consistent with the mtDNA sequences and STR profiles. Allelic dropout occurred in some urine samples that were examined for STR analysis. Case-type sample studies demonstrated that mtDNA testing could be used to obtain DNA profiles of all the samples tested, including blood, dried blood spots, urine, blood residues on needles, and blood stains. In conclusion, mtDNA testing is valuable for analyzing highly degraded DNA samples, such as urine samples, compared with STR analysis. Urine testing should be performed for the initial testing procedure, because mtDNA is inherited maternally. In situations where the DNA match is detrimental to the athlete, additional blood STR analysis may be required.

Top-Down Proteomics Analysis of Picogram-Level Complex Samples Using Spray-Capillary-Based Capillary Electrophoresis-Mass Spectrometry.

Proteomics analysis of mass-limited samples has become increasingly important for understanding biological systems in physiologically relevant contexts such as patient samples, multicellular organoids, spheroids, and single cells. However, relatively low sensitivity in top-down proteomics methods makes their application to mass-limited samples challenging. Capillary electrophoresis (CE) has emerged as an ideal separation method for mass-limited samples due to its high separation resolution, ultralow detection limit, and minimal sample volume requirements. Recently, we developed "spray-capillary", an electrospray ionization (ESI)-assisted device, that is capable of quantitative ultralow-volume sampling (e.g., pL-nL level). Here, we developed a spray-capillary-CE-MS platform for ultrasensitive top-down proteomics analysis of intact proteins in mass-limited complex biological samples. Specifically, to improve the sensitivity of the spray-capillary platform, we incorporated a polyethylenimine (PEI)-coated capillary and optimized the spray-capillary inner diameter. Under optimized conditions, we successfully detected over 200 proteoforms from 50 pg of E. coli lysate. To our knowledge, the spray-capillary CE-MS platform developed here represents one of the most sensitive detection methods for top-down proteomics. Furthermore, in a proof-of-principle experiment, we detected 261 ± 65 and 174 ± 45 intact proteoforms from fewer than 50 HeLa and OVCAR-8 cells, respectively, by coupling nanodroplet-based sample preparation with our optimized CE-MS platform. Overall, our results demonstrate the capability of the modified spray-capillary CE-MS platform to perform top-down proteomics analysis on picogram amounts of samples. This advancement presents the possibility of meaningful top-down proteomics analysis of mass-limited samples down to the level of single mammalian cells.

Online Nucleotide Mapping of mRNAs.

Messenger RNA (mRNA) can be sequenced via indirect approaches such as Sanger sequencing and next generation sequencing (NGS), or direct approaches like bottom-up mass spectrometry (MS). Direct sequencing allows the confirmation of RNA modifications. However, the conventional bottom-up MS approach involves time-consuming in-solution digestions that require a large amount of sample, and can lead to the RNase contamination of the LC-MS system and column. Here, we describe a platform that enables online nucleotide mapping of mRNAs via the use of immobilized RNase cartridges and 2D-LC-MS instrumentation. The online approach was compared to conventional offline digestion protocols adapted from two published studies. For this purpose, five model mRNAs of varying lengths (996-4521 nucleotides) and chemistries (unmodified uridine vs 5-methoxyuridine (5moU)) were analyzed. The profiles and sequence coverages obtained after RNase T1 digestion were discussed. The online nucleotide mapping achieved comparable or slightly greater sequence coverage for the 5 mRNAs (5.8-51.5%) in comparison to offline approaches (3.7-50.4%). The sequence coverage was increased to 65.6-85.6 and 69.7-85.0% when accounting for the presence of nonunique digestion products generated by the RNase T1 and A, respectively. The online nucleotide mapping significantly reduced the digestion time (from 15 to <5 min), increased the signal intensity by more than 10-fold in comparison to offline approaches.

Development of a Multiplexing Injector for Gas Chromatography for the Time-Resolved Analysis of Volatile Emissions from Lithium-Ion Batteries.

Multiplex sampling, so far mainly used as a tool for S/N ratio improvement in spectroscopic applications and separation techniques, has been investigated here for its potential suitability for time-resolved monitoring where chromatograms of transient signals are recorded at intervals much shorter than the chromatographic runtime. Different designs of multiplex sample introduction were developed and utilized to analyze lithium-ion battery degradation products under normal or abuse conditions to achieve fast and efficient sample introduction. After comprehensive optimization, measurements were performed on two different GC systems, with either barrier discharge ionization detection (BID) or mass spectrometric detection (MS). Three different injector designs were examined, and modifications in the pertinent hardware components and operational conditions used. The shortest achievable sample introduction time was 50 ms with an interval of 6 s. Relative standard deviations were lower than 4% and 10% for the intra- and inter-day repeatability, respectively. The sample introduction system and column head pressure had to be carefully controlled, as this parameter most critically affects the amount of sample introduced and, thus, detector response. The newly developed sample introduction system was successfully used to monitor volatile degradation products of lithium-ion batteries and demonstrated concentration changes over the course of time of the degradation products (e.g., fluoroethane, acetaldehyde and ethane), as well as for solvents from the battery electrolyte like ethyl carbonate.

Highly sensitive and label-free detection of naproxen using mixed metal oxide-based field effect transistor as a biosensor for in-vitro analysis of urine

Naproxen (Nap) is a nonsteroidal anti-inflammatory drug widely prescribed for treating fever, inflammation, and pain due to its easy absorption. Considering the side effects of long-term use of Nap and the risk of environmental pollution as a precursor, it is necessary to measure its concentration. ISFET-based sensors are very attractive tools for monitoring small amounts of analytes in biological samples. On the other hand, stabilizing nanocomposites with Nap oxidation capability provides an electrochemical signal for quantification. In this study, ISFET was used for comparative analysis of Nap concentration employing CeO2/CuO/MnO, CeO2/CuO/NiO multi-metal oxides, and La-doped CoFe layered double hydroxide (LDH) as sensing layers for the first time. The detection limits of the mentioned layers were 14.08 μM, 11.15 μM, and 19.11 μM, respectively, and the method was used to detect Nap in urine with a linear range of 0–400 μM. Compared to other methods, the proposed ISFET-based sensor for Nap detection has the capability of miniaturization, low sample amount, real-time measurement, low electrical power requirement, reusability, and high stability.

Isolation of Mitochondria for Mitochondrial Supercomplex Analysis from Small Tissue and Cell Culture Samples.

Over the last decades, the evidence accumulated about the existence of respiratory supercomplexes (SCs) has changed our understanding of the mitochondrial electron transport chain organization, giving rise to the proposal of the "plasticity model." This model postulates the coexistence of different proportions of SCs and complexes depending on the tissue or the cellular metabolic status. The dynamic nature of the assembly in SCs would allow cells to optimize the use of available fuels and the efficiency of electron transfer, minimizing reactive oxygen species generation and favoring the ability of cells to adapt to environmental changes. More recently, abnormalities in SC assembly have been reported in different diseases such as neurodegenerative disorders (Alzheimer's and Parkinson's disease), Barth Syndrome, Leigh syndrome, or cancer. The role of SC assembly alterations in disease progression still needs to be confirmed. Nevertheless, the availability of enough amounts of samples to determine the SC assembly status is often a challenge. This happens with biopsy or tissue samples that are small or have to be divided for multiple analyses, with cell cultures that have slow growth or come from microfluidic devices, with some primary cultures or rare cells, or when the effect of particular costly treatments has to be analyzed (with nanoparticles, very expensive compounds, etc.). In these cases, an efficient and easy-to-apply method is required. This paper presents a method adapted to obtain enriched mitochondrial fractions from small amounts of cells or tissues to analyze the structure and function of mitochondrial SCs by native electrophoresis followed by in-gel activity assays or western blot.

A Fully Automated Online Enrichment and Separation System for Highly Reproducible and In-Depth Analysis of Intact Glycopeptide.

A fully automated online enrichment and separation system for intact glycopeptides, named AutoGP, was developed in this study by integrating three different columns in a nano-LC system. Specifically, the peptide mixture from the enzymatic digestion of a complex biological sample was first loaded on a hydrophilic interaction chromatography (HILIC) column. The nonglycopeptides in the sample were washed off the column, and the glycopeptides retained by the HILIC column were eluted to a C18 trap column to achieve an automated glycopeptide enrichment. The enriched glycopeptides were further eluted to a C18 column for separation, and the separated glycopeptides were eventually analyzed by using an orbitrap mass spectrometer (MS). The optimal operating conditions for AutoGP were systemically studied, and the performance of the fully optimized AutoGP was compared with a conventional manual system used for glycopeptide analysis. The experimental evaluation shows that the total number of glycopeptides identified is at least 1.5-fold higher, and the median coefficient of variation for the analyses is at least 50% lower by using AutoGP, as compared to the results acquired by using the manual system. In addition, AutoGP can perform effective analysis even with a 1-μg sample amount, while a 10-μg sample at least will be needed by the manual system, implying an order of magnitude better sensitivity of AutoGP. All the experimental results have consistently proven that AutoGP can be used for much better characterization of intact glycopeptides.

Nonparametric Modelling of Ship Dynamics Using Puma Optimizer Algorithm-Optimized Twin Support Vector Regression

Ship dynamic models serve as the foundation for designing ship controllers, trajectory planning, and obstacle avoidance. Support vector regression (SVR) is a commonly used nonparametric modelling method for ship dynamics. Achieving high accuracy SVR models requires a substantial amount of training samples. Additionally, as the number of training samples increases, the computational efficiency for solving the quadratic programming problem (QPP) of SVR decreases. Ship controllers demand dynamic models with both high accuracy and computational efficiency. Therefore, to enhance the prediction accuracy and computational efficiency of SVR, this paper proposes a nonparametric modelling method based on twin SVR (TSVR). TSVR replaces a large QPP with a set of smaller QPPs, significantly enhancing generalizability and computational efficiency. To further improve the predictive accuracy of TSVR, the puma optimizer algorithm is employed to determine the optimal hyperparameters. The performance of the proposed method is validated using a Mariner class vessel. Gaussian white noise is introduced into the modelling data to simulate measurement error. The TSVR model accurately predicts various zigzag and turning circle manoeuvring motions under disturbance conditions, demonstrating its robustness and generalizability. Compared to the SVR model, the TSVR model achieves lower root mean square error and computational time, confirming its superior predictive accuracy and computational efficiency.

A 10-year follow-up study on clinical outcomes of dental implant rehabilitation using surgical guide.

The surgical guide is a static computer-assisted device used for implant surgery planning and guidance. By taking an impression and referring to the patients' three-dimensional computed tomography scan of the desired implant site, a surgical guide can be created. During surgery, the surgical guide aids in achieving the designed implant placement position and direction. We examined and evaluated the long-term clinical outcomes of implant surgery using surgical guides. This study investigated a total of 15 patients with 32 implants that were placed using surgical guides from 2009 to 2011 with a mean follow-up period extended beyond 10 years. Patient demographics and implant survival rates were recorded. We analyzed marginal bone loss (MBL) by assessing the radiographs acquired at installation, three months after installation, and one month, one, two, and five years after prosthesis delivery. The mean patient age was 57.33 years at implant placement. Of the 32 implants, five implants were placed in the anterior region and 27 implants were in the posterior region. Six implants failed and three of them were replaced, resulting in an 81.25% survival rate. The mean follow-up period was 10 years and nine months. Mean MBL compared to post-installation was significantly higher than at three months after installation, and one month, one, two, and five years after prosthesis delivery. Mean MBL at three months after installation, and one month, one year, and two years were significantly higher compared to the previous visit (P<0.05). However, MBL at five years after prosthesis delivery did not differ significantly compared to at two years. In this study, implant rehabilitation assisted by surgical guides exhibited favorable survival rates. With the limitation of the sample amount in this study, further research and more samples are required to evaluate the long-term clinical effectiveness of surgical guides.