• Mass Spectrometry Letters
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• v.5 no.1
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• pp.1-11
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• 2014

Lipids play important roles in biological systems; they store energy, play a structural role in the cell membrane, and are involved in cell growth, signal transduction, and apoptosis. Phospholipids (PLs) in particular have received attention in the medical and lipidomics research fields because of their involvement in human diseases such as diabetes, obesity, atherosclerosis, and many cancers associated with lipid metabolic disorders. Here I review experimental strategies for PL analysis based on nanoflow liquid chromatography-electrospray ionization-tandem mass spectrometry (nLC-ESI-MSn). In particular, discussed are lipid extraction methods, nanoflow LC separation of PLs, effect of ionization modifiers on the ESI of PLs, influence of chain lengths and unsaturation degree of acyl chains of PLs on MS intensity, structural determination of the molecular structure of PLs and their oxidized products, and quantitative profiling of PLs from biological samples such as tissue, urine, and plasma in relation to cancer and coronary artery disease.

• Membrane Journal
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• v.26 no.4
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• pp.291-300
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• 2016

Techniques for selectively separating molecules of gas and liquid states using various separation membranes have been widely used in variety of applications such as chemical, biological, pharmaceutical, and petrochemical industries. As the nanochannel diameter, inter-channel distance and length of the nanochannel of the anodic aluminum oxide (AAO) membranes can be precisely controlled, various studies to effectively separate mixture of various molecules using AAO membrane have been widely carried out. In this study, we fabricated AAO membranes of cylindrical nanochannels of various diameter sizes and of through-hole structure, that is, nanochannels of which both ends of each nanochannel are open. Using those AAO membranes of through-hole nanochannel structure, we studied the selective permeation polymer chains dissolved in a solvent based on hydraulic volume of the polymer chains. We found a precise, quantitative relationship between the radius of gyration of polymer chains that permeated through nanochannels inside AAO membrane and the diameter of nanochannels. In addition, we demonstrate that the behavior of the polymer solution flowing through nanochannel of the AAO membrane can be successfully described with the Hagen-Poiseuille relationship. It is, therefore, possible to theoretically interpret the nanoflow of the solution flowing inside the cylindrical nanochannel.

• Mass Spectrometry Letters
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• v.2 no.3
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• pp.65-68
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• 2011

The effects of column length and particle size on the efficiency of separation and characterization of phospholipids (PLs) are investigated using nanoflow liquid chromatography-electrospray ionization-tandem mass spectrometry (nLC-ESI-MS-MS). Since PLs are associated with cell proliferation, apoptosis, and signal transduction, it is of increasing interests in lipidomics to establish reliable analytical methods for the qualitative and quantitative profiling of PLs related to biomarker development in adult diseases. Due to the complexity of PLs, the preliminary separation of PLs is necessary prior to MS analysis. In this study, length of capillary column and the particle size of reversed phase ($C_{18}$) packing materials are varied to find a reliable condition for the high speed and high resolution separation using 8 PL standard mixtures. From experiments, it was found that a capillary column of nLC-ESI-MS-MS analysis for PL mixtures can be minimized to a 5 cm long pulled tip column packed with 3 ${\mu}m$ $C_{18}$ particles without losing resolution.

• Mass Spectrometry Letters
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• v.1 no.1
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• pp.21-24
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• 2010

The development of clinical biomarkers involves discovery, verification, and validation. Recently, multiple reaction monitoring (MRM) coupled with stable isotope dilution mass spectrometry (IDMS) has shown considerable promise for the direct quantification of proteins in clinical samples. In particular, multiple biomarkers have been tracked in a single experiment using MRM-based MS approaches combined with liquid chromatography. We report here a highly reproducible, quantitative, and dynamic MRM system for validating multi-biomarker proteins using Nanoflow HPLC-Microfluidics Chip/Triple-Quadrupole MS. In this system, transitions were acquired only during the retention window of each eluting peptide. Transitions with the highest MRM-MS intensities for the five target peptides from colon cancer biomarker candidates were automatically selected using Optimizer software. Relative to the corresponding non-dynamic system, the dynamic MRM provided significantly improved coefficients of variation in experiments with large numbers of transitions. Linear responses were obtained with concentrations ranging from fmol to pmol for five target peptides.

• Journal of the Semiconductor & Display Technology
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• v.1 no.1
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• pp.29-33
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• 2002

In the flip-chip process, the problem like electric defect or fatigue crack caused by the difference of CTE, between chip and substrate board had occurred. Underfill of flip chip to overcome this defects is noticed as important work developing in whole reliability of chip by protecting the chip against the external shock. In this paper, we introduce the underfill methods using mold and plunge and improvement of process and reliability, and the advantage which can be taken from embodiment of device.

• Molecular & Cellular Toxicology
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• v.1 no.2
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• pp.87-91
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• 2005

Although 2D-PAGE has been widely used as the primary method for protein separation, difficulties in displaying proteins with an extreme values of isoelectric paint (pI), molecular size and hydrophobicity limit the technique. In addition, time consuming steps involving protein transfer and extraction from the gel-pieces can result in sample loss. Here, we describe a novel protein separation technique with capillary size-exclusion chromatography (CSEC) for rapid protein identification from human plasma. The method includes protein fractionation along with molecular size followed by in-solution tryptic digestion and peptide analysis through reversed phase liquid chromatography (RPLC) coupled to nanoflow electrospray-tandem mass spectrometry (ESI-MS/MS). Tryptic peptides are applied an a $100\;{\mu}m\;i.d.{\times}10mm$ length pre-column and then separated on a $75\;{\mu}m{\times}200mm$ analytical column at -100 nL/min flaw rate. Proteins were identified over the wide ranges of pI (3.7-12.3) when this technique was applied to the analysis of $1-2\;{\mu}L$ of human plasma. This gel-free system provides fast fractionation and may be considered a complementary technique to SDS-PAGE in proteomics.

• Bulletin of the Korean Chemical Society
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• v.35 no.4
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• pp.1133-1138
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• 2014

Ultrahigh performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI- MS/MS) provides a high-speed method to screen a large number of samples for small molecules with specific properties. In this study, UPLC-ESI-MS/MS with multiple reaction monitoring (MRM) was employed to screen urinary phospholipid (PL) content for biomarkers of prostate cancer. From lists of urinary PLs structurally identified using nanoflow LC-ESI-MS/MS, 52 PL species were selected for quantitative analysis in urine samples between 22 cancer-free urologic patients as controls and 45 prostate cancer patients. Statistical treatment of data by receiver operating characteristic (ROC) analysis yielded 14 PL species that differed significantly in relative concentrations (area under curve (AUC) > 0.8) between the two groups. Among PLs present at higher levels in prostate cancer urine, phosphatidylcholines (PCs) and phosphatidylinositols (PIs) constituted the major head group PLs (3 PCs and 7 PIs). For technical reasons, PL species of low abundance may be underrepresented in data from UPLC-ESI-MS/MS performed in MRM mode. However, the proposed method enables the rapid screening of large numbers of plasma or urine samples in the search for biomarkers of human disease.

• Restorative Dentistry and Endodontics
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• v.49 no.2
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• pp.20.1-20.13
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• 2024

Objectives: This study investigated the nanoleakage of root canal obturations using calcium silicate-based sealer according to different drying methods. Materials and Methods: Fifty-two extracted mandibular premolars with a single root canal and straight root were selected for this study. After canal preparation with a nickel-titanium rotary file system, the specimens were randomly divided into 4 groups according to canal drying methods (1: complete drying, 2: blot drying/distilled water, 3: blot drying/NaOCl, 4: aspiration only). The root canals were obturated using a single-cone filling technique with a calcium silicate-based sealer. Nanoleakage was evaluated using a nanoflow device after 24 hours, 1 week, and 1 month. Data were collected twice per second at the nanoscale and measured in nanoliters per second. Data were statistically analyzed using the Kruskal-Wallis and Mann-Whitney U-tests (p < 0.05). Results: The mean flow rate measured after 24 hours showed the highest value among the time periods in all groups. However, the difference in the flow rate between 1 week and 1 month was not significant. The mean flow rate of the complete drying group was the highest at all time points. After 1 month, the mean flow rate in the blot drying group and the aspiration group was not significantly different. Conclusions: Within the limitations of this study, the canal drying method had a significant effect on leakage and sealing ability in root canal obturations using a calcium silicate-based sealer. Thus, a proper drying procedure is critical in endodontic treatment.

• IMMUNE NETWORK
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• v.23 no.4
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• pp.28.1-28.20
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• 2023

Lipid accumulation in macrophages is a prominent phenomenon observed in atherosclerosis. Previously, intimal foamy macrophages (FM) showed decreased inflammatory gene expression compared to intimal non-foamy macrophages (NFM). Since reprogramming of lipid metabolism in macrophages affects immunological functions, lipid profiling of intimal macrophages appears to be important for understanding the phenotypic changes of macrophages in atherosclerotic lesions. While lipidomic analysis has been performed in atherosclerotic aortic tissues and cultured macrophages, direct lipid profiling has not been performed in primary aortic macrophages from atherosclerotic aortas. We utilized nanoflow ultrahigh-performance liquid chromatography-tandem mass spectrometry to provide comprehensive lipid profiles of intimal non-foamy and foamy macrophages and adventitial macrophages from Ldlr^-/- mouse aortas. We also analyzed the gene expression of each macrophage type related to lipid metabolism. FM showed increased levels of fatty acids, cholesterol esters, phosphatidylcholine, lysophosphatidylcholine, phosphatidylinositol, and sphingomyelin. However, phosphatidylethanolamine, phosphatidic acid, and ceramide levels were decreased in FM compared to those in NFM. Interestingly, FM showed decreased triacylglycerol (TG) levels. Expressions of lipolysis-related genes including Pnpla2 and Lpl were markedly increased but expressions of Lpin2 and Dgat1 related to TG synthesis were decreased in FM. Analysis of transcriptome and lipidome data revealed differences in the regulation of each lipid metabolic pathway in aortic macrophages. These comprehensive lipidomic data could clarify the phenotypes of macrophages in the atherosclerotic aorta.