• Biomedical Science Letters
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• v.25 no.4
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• pp.426-430
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• 2019

Currently, molecular diagnostic assays based on nucleic acid amplification tests have been shown to effectively detect mycobacterial infections in various types of specimen, however, variable sensitivity was shown in FFPE samples according to the kind of commercial kit used. The present study therefore used automated PCR-reverse blot hybridization assay (REBA) system, REBA Myco-ID HybREAD 480^®, for the rapid identification of Mycobacterium species in various types of human tissue and compared the conventional one-tube nested-PCR assay for detecting Mycobacterium tuberculosis (MTB). In conventional nested-PCR tests, 25 samples (48%) were MTB positive and 27 samples (52%) were negative. In contrast, when conducted PCR-REBA assay, 11 samples (21%) were MTB positive, 20 samples (39%) were NTM positive, 8 samples (15%) were MTB-NTM double positive, and 13 samples (25%) were negative. To determine the accuracy and reliability of the two molecular diagnostic tests, the one-tube nested-PCR and PCR-REBA assays, were compared with histopathological diagnosis in discordant samples. When conducted nested-PCR assay, 10 samples (59%) were MTB positive and seven samples (41%) were negative. In contrast, when conducted PCR-REBA test, three samples (17%) were MTB positive, 10 samples (59%) were NTM positive and four samples (24%) were negative. In conclusion, the automated PCR-REBA system proved useful to identify Mycobacterium species more rapidly and with higher sensitivity and specificity than the conventional molecular assay, one-tube nested-PCR; it might therefore be the most suitable tool for identifying Mycobacterium species in various types of human tissue for precise and accurate diagnosis of mycobacterial infection.

• Genomics & Informatics
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• v.4 no.4
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• pp.161-166
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• 2006

The establishment of DNA microarray technology has enabled high-throughput analysis and molecular profiling of various types of cancers. By using the gene expression data from microarray analysis we are able to investigate diagnostic applications at the molecular level. The most important step in the application of microarray technology to cancer diagnostics is the selection of specific markers from gene expression profiles. In order to select markers of Immortalization and transformation we used c-myc and $H-ras^{V12}$ oncogene-transfected NIH3T3 cells as our model system. We have identified 8751 differentially expressed genes in the immortalization/transformation model by multivariate permutation F-test (95% confidence, FDR<0.01). Using the support vector machine algorithm, we selected 13 discriminative genes which could be used to predict immortalization and transformation with perfect accuracy. We assayed $H-ras^{V12}$-transfected 'transformed' cells to validate our immortalization/transformation dassification system. The selected molecular markers generated valuable additional information for tumor diagnosis, prognosis and therapy development.

• Journal of Veterinary Science
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• v.23 no.3
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• pp.50.1-50.12
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• 2022

Background: There is an urgent need to find reliable and rapid bovine tuberculosis (bTB) diagnostics in response to the rising prevalence of bTB worldwide. Toll-like receptor 2 (TLR2) recognizes components of bTB and initiates antigen-presenting cells to mediate humoral immunity. Evaluating the affinity of antigens with TLR2 can form the basis of a new method for the diagnosis of bTB based on humoral immunity. Objectives: To develop a reliable and rapid strategy to improve diagnostic tools for bTB. Methods: In this study, we expressed and purified the sixteen bTB-specific recombinant proteins in Escherichia coli. The two antigenic proteins, MPT70 and MPT83, which were most valuable for serological diagnosis of bTB were screened. Molecular docking technology was used to analyze the affinity of MPT70, MPT83, dominant epitope peptide of MPT70 (M1), and dominant epitope peptide MPT83 (M2) with TLR2, combined with the detection results of enzyme-linked immunosorbent assay to evaluate the molecular docking effect. Results: The results showed that interaction surface Cα-atom root mean square deviation of proteins (M1, M2, MPT70, MPT83)-TLR2 protein are less than 2.5 A, showing a high affinity. It is verified by clinical serum samples that MPT70, MPT83, MPT70-MPT83 showed good diagnostic potential for the detection of anti-bTB IgG and M1, M2 can replace the whole protein as the detection antigen. Conclusions: Molecular docking to evaluate the affinity of bTB protein and TLR2 combined with ELISA provides new insights for the diagnosis of bTB.

• Proceedings of the KWS Conference
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• 2002.10a
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• pp.612-619
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• 2002

The dynamic behavior of Al-Mg alloys plasma was very unstable and this instability was closely related to the unstable motion of keyhole during laser irradiation. The keyhole fluctuated both in size and shape and its fluctuation period was about 440 ${\mu}{\textrm}{m}$. This instability has been estimated to be caused by the evaporation phenomena of metals with different boiling point and latent heats of vaporization. Therefore, the authors have conducted the spectroscopic diagnostics of plasma induced in the pulsed YAG laser welding of Al-Mg alloys in air and argon atmospheres. In the air environment, the identified spectra were atomic lines of Al, Mg, Cr, Mn, Cu, Fe and Zn, and singly ionized Mg line, as well as strong molecular spectrum of AlO, MgO and AIH. It was confirmed that the resonant lines of Al and Mg were strongly self-absorbed, in particular in the vicinity of pool surface. The self-absorption of atomic Mg line was more eminent in alloys containing higher Mg. These facts showed that the laser-induced plasma was relatively a low temperature and high density metallic vapor. The intensities of molecular spectra of AlO and MgO were different each other depending on the power density of laser beam. Under the low power density irradiation condition, the MgO band spectra were predominant in intensity, while the AlO spectra became much stronger in higher power density. In argon atmosphere the band spectra of MgO and AlO completely vanished, but AlH molecular spectra was detected clearly. The hydrogen source was presumably the hydrogen solved in the base Metal, absorbed water on the surface oxide layer or H$_2$ and $H_2O$ in the shielding gas. The temporal change in spectral line intensities was quite similar to the fluctuation of keyhole. The time average plasma temperature at 1 mm high above the surface of A5083 alloy was determined by the Boltzmann plot method of atomic Cr lines of different excitation energy. The obtained electron temperature was 3, 280$\pm$150 K which was about 500 K higher than the boiling point of pure aluminum. The electron number density was determined by measuring the relative intensities of the spectra1lines of atomic and singly ionized Magnesium, and the obtained value was 1.85 x 1019 1/㎥.

• Korean Journal of Clinical Laboratory Science
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• v.51 no.1
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• pp.26-33
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• 2019

Recently, the rapid growth of the companion animal market has led to the development of animal disease diagnosis kits. Therefore, the utility of the introduction of biomarkers for the development of animal molecular diagnostics is being reevaluated. A good biomarker should be precise and reliable, distinguish between normal and diseased states, and differentiate between different diseases. Recently reported genetic markers, tumor markers (cell free DNA, circulating tumor cells, granzyme, and skin tumors), and others (brucellosis, programmed death recovery-1, symmetric dimethylarginine, periostin, and cysteinyl leukotrien) have been developed. The biomarkers are used for risk prediction or for the screening, diagnosis, and monitoring of disease progression. The most important criteria for related biomarkers are disease specificity. Many potential biomarkers have emerged from laboratory and test studies, but they have not been validated in independent or large-scale clinical studies. Candidate biomarkers evaluate disease associations, verify the effectiveness of biomarkers for early detection and disease progression, and incorporate them into humans and animals. In the future, it will be necessary to reevaluate the utility of well-structured biomarker-based research and study the development of kits that can be used in on-site tests in accordance with the trends introduced in the diagnosis of animal diseases.

• Clinical and Experimental Pediatrics
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• v.55 no.3
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• pp.77-82
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• 2012

Human astrovirus (HAstV) is a major cause of acute diarrhea among children, resulting in outbreaks of diarrhea and occasionally hospitalization. Improved surveillance and application of sensitive molecular diagnostics have further defined the impact of HAstV infections in children. These studies have shown that HAstV infections are clinically milder (diarrhea, vomiting, fever) than infections with other enteric agents. Among the 8 serotypes of HAstV identified, serotype 1 is the predominant strain worldwide. In addition to serotype 1, the detection rate of HAstV types 2 to 8 has increased by using newly developed assays. HAstV is less common compared with other major gastroenteritis viruses, including norovirus and rotavirus; however, it is a potentially important viral etiological agent with a significant role in acute gastroenteritis. A better understanding of the molecular epidemiology and characteristics of HAstV strains may be valuable to develop specific prevention strategies.

• BMB Reports
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• v.44 no.12
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• pp.765-771
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• 2011

Cancer treatment has been stratified by companion biomarker tests that serve to provide information on the genetic status of cancer patients and to identify patients who can be expected to respond to a given treatment. This stratification guarantees better efficiency and safety during treatment. Cancer patients, however, marginally benefit from the current companion biomarker-aided treatment regimens, presumably because companion biomarker tests are dependent solely on the mutation status of several genes status quo. In the true sense of the term, "personalized medicine", cancer patients are deemed to be identified individually by their molecular signatures, which are not necessarily confined to genetic mutations. Glycosylation is tremendously dynamic and shows alterations in cancer. Evidence is accumulating that aberrant glycosylation contributes to the development and progression of cancer, holding the promise for use of glycosylation status as a companion biomarker in cancer treatment. There are, however, several challenges derived from the lack of a reliable detection system for aberrant glycosylation, and a limited library of aberrant glycosylation. The challenges should be addressed if glycosylation status is to be used as a companion biomarker in cancer treatment and contribute to the fulfillment of personalized medicine.

• Proceedings of the Korean Vacuum Society Conference
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• 2011.02a
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• pp.15-15
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• 2011

Atom-thick graphene membrane and nano-sized graphene objects (NGOs) hold substantial potential for applications in future molecular-scale integrated electronics, transparent conducting membranes, nanocomposites, etc. To realize this potential, chemical properties of graphene need to be understood and diagnostic methods for various NGOs are also required. To meet these needs, chemical properties of graphene and optical diagnostics of graphene nanoribbons (GNRs) have been explored by Raman spectroscopy, AFM and STM scanning probes. The first part of the talk will illustrate the role of underlying silicon dioxide substrates and ambient gases in the ubiquitous hole doping of graphene. An STM study reveals that thermal annealing generates out-of-plane deformation of nanometer-scale wavelength and distortion in $sp^2$ bonding on an atomic scale. Graphene deformed by annealing is found to be chemically active enough to bind molecular oxygen, which leads to a strong hole-doping. The talk will also introduce Raman spectroscopy studies of GNRs which are known to have nonzero electronic bandgap due to confinement effect. GNRs of width ranging from 15 nm to 100 nm have been prepared by e-beam lithographic patterning of mechanically exfoliated graphene followed by oxygen plasma etching. Raman spectra of narrow GNRs can be characterized by upshifted G band and strong disorder-related D band originating from scattering at ribbon edges. Detailed analysis of the G, D, and 2D bands of GNRs proves that Raman spectroscopy is still a reliable tool in characterizing GNRs despite their nanometer width.

• Clinical and Experimental Pediatrics
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• v.63 no.6
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• pp.195-202
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• 2020

Developments in next-generation sequencing (NGS) techogies have assisted in clarifying the diagnosis and treatment of developmental delay/intellectual disability (DD/ID) via molecular genetic testing. Advances in DNA sequencing technology have not only allowed the evolution of targeted panels but also, and more currently enabled genome-wide analyses to progress from research era to clinical practice. Broad acceptance of accuracy-guided targeted gene panel, whole-exome sequencing (WES), and whole-genome sequencing (WGS) for DD/ID need prospective analyses of the increasing cost-effectiveness versus conventional genetic testing. Choosing the appropriate sequencing method requires individual planning. Data are required to guide best-practice recommendations for genomic testing, regarding various clinical phenotypes in an etiologic approach. Targeted panel testing may be recommended as a firsttier testing approach for children with DD/ID. Family-based trio testing by WES/WGS can be used as a second test for DD/ID in undiagnosed children who previously tested negative on a targeted panel. The role of NGS in molecular diagnostics, treatment, prediction of prognosis will continue to increase further in the coming years. Given the rapid pace of changes in the past 10 years, all medical providers should be aware of the changes in the transformative genetics field.

• Design & Manufacturing
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• v.11 no.3
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• pp.35-40
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• 2017

PCR(Polymerase chain reaction) is a technique to replicate and amplify a desired part of DNA. It is used in various aspects such as DNA fingerprint analysis and rare DNA amplification of an extinct animal. Especially in the medical diagnosis field, it provides various measurement methods at the molecular level such as genetic diagnosis, and is a basic tool for molecular diagnostics. The internal shape of the plastic vial tube for PCR analysis used in the DNA detection process, and the surface roughness and internal cleanliness can affect detection and discrimination results. The plastic vial tube demanded by the developer of the PCR analysis equipment should be changed to a structure that eliminates the residual washing solution in the washing process to ensure the internal cleanliness. Thus the internal structure and the internal surface design for improving the PCR amplification efficiency are key issues to develop the plastic vial tube for the DNA detection process.