• Title/Summary/Keyword: molecular biological techniques

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Recent Advances in Nuclear Medicine Imaging Instrumentation (핵의학 영상기기의 최근 진보)

  • Jung, Jin-Ho;Choi, Yong;Hong, Key-Jo;Min, Byung-Jun;Hu, Wei;Kang, Ji-Hoon
    • Nuclear Medicine and Molecular Imaging
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    • v.42 no.2
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    • pp.98-111
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    • 2008
  • This review introduces advances in clinical and pre-clinical single photon emission computed tomography (SPECT) and positron emission tomography (PET) providing noninvasive functional images of biological processes. Development of new collimation techniques such as multi-pinhole and slit-slat collimators permits the improvement of system spatial resolution and sensitivity of SPECT. Application specific SPECT systems using smaller and compact solid-state detector have been customized for myocardial perfusion imaging with higher performance. Combined SPECT/CT providing improved diagnostic and functional capabilities has been introduced. Advances in PET and CT instrumentation have been incorporated in the PET/CT design that provide the metabolic information from PET superimposed on the anatomic information from CT. Improvements in the sensitivity of PET have achieved by the fully 3D acquisition with no septa and the extension of axial field-of-view. With the development of faster scintillation crystals and electronics, time-of-flight (TOF) PET is now commercially available allowing the increase in the signal-to-noise ratio by incorporation of TOF information into the PET reconstruction process. Hybrid PET/SPECT/CT systems has become commercially available for molecular imaging in small animal models. The pre-clinical systems have improved spatial resolution using depth-of-interaction measurement and new collimators. The recent works on solid state detector and dual modality nuclear medicine instrumentations incorporating MRI and optical imagers will also be discussed.

Purification and Characterization of Storage Proteins from the Mulberry Longicorn Beetle, Apriona germari Hope

  • Yoon, Hyung-Joo;Kim, Seong-Ryul;Jin, Byung-Rae;Lee, Sang-Mong;Moon, Jae-Yu;Mah, Young-Il;Soh, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
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    • v.2 no.2
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    • pp.161-166
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    • 2001
  • The storage proteins of the mulberry longicorn beetle, Apriona germari Hope, were purified and characterized. Three kinds of storage protein (SP1, SP2 and Sp3) were purified from the last instar larval hemolymph of A. germari by the FPLC techniques, anion exchange chromatography and gel permeation chromatography. The SP1, SP2 and SP3 have a native molecular weight of 480, 440 and 420 kDa, respectively. In the SDS-polyacrylamide gel electrophoresis analysis, these storage proteins are composed of a single protein subunit with molecular weight of 90, 85 and 80 kDa, respectively. This result showed that the storage proteins are hexameric protein. The SP1 and SP2 were stained with Schiffs reagent, but SP3 was not stained. It can be assumed that SP1 and SP2 are glycoprotein. Western blot analyses using the each of polyclonal antiserum against purified SP1, SP2 and SP3 showed that the three antibodies reacted with the each of SP bands, respectively. Also, antibodies against SP1 and SP3 cross-reacted with the SP3 and SP1, respectively. However, SP2 was not cross-reacted with these two antibodies. Also, antiserum against SP2 did not cross-reacted with the SP1 and SP3.

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Development of Novel Pyrrolidine Organocatalyst

  • Im, Seol-Hui;Gang, Seong-Ho
    • Proceedings of the Korean Vacuum Society Conference
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    • 2011.08a
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    • pp.198-198
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    • 2011
  • Organocatalysis is a relatively new and popular area within the field of chiral molecule synthesis. It is one of the main branches of enantioselective synthesis with enzymatic and organometallic catalysis. In recent years, immense high quality studies on catalysis by chiral secondary amines were reported. These progresses instantly led to different organocatalytic activation concepts, so thousands of researchers from academia and the chemical industry are currently involved in this field and new ideas, new approaches, and creative thinking have been rapidly emerged. Organocatalysts, some of which are natural products, appear to solve the problems of metal catalysts. Compared to metal-based catalysis, they have many advantages including savings in cost, time, and energy, easier experimental procedure, and reduction of chemical waste. These benefits originate from the following factors. First, organocatalysts are generally stable in oxygen and water in the atmosphere, there is no need for special equipments or experimental techniques to operate under anhydrous or anaerobic conditions. Second, organic reagents are naturally available from biological materials as single enantiomers that they are easy and cheap to prepare which makes them suitable for small-scale to industrial-scale reactions. Third, in terms of safety related catalysis, small organic molecules are non-toxic and environmentally friendly. Therefore, the purpose of this research is to develop novel synthetic methods and design for various organocatalyst. Furthermore, it is expected that these organocatalysts can be applied to a variety of asymmetric reactions and study the transition state of these reactions using a metal sulface. Here, we report the synthesis of unprecedented organocatalysts, proline and pyrrolidine derivatives with quaternary carbon center.

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Solubility and Electrophoretic pattern of Korea Ginseng Protein (한국산 인삼 단백질의 용해성 및 전기영동 패턴)

  • Choi, Cheong;Yoon, Sang-Hong;Bae, Man-Jong;An, Bong-Jeon
    • Applied Biological Chemistry
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    • v.28 no.2
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    • pp.88-91
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    • 1985
  • For the systematic investigation of biochemical characteristics of Korean ginseng protein, protein fractions were analyzed by the techniques of sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. The effect of pH and various salts on extractibility of ginseng protein were determined while the amino acid composition was studied by amino acid autoanalyzer. The protein was consisted of 66.08% of albumin and 20.51% of glutelin. Extractability of ginseng protein was the lowest in pH 3.0 and the highest in $pH\;6.0{\sim}8.0$. Among the neutral salts solution, $0.4M\;Na_2CO_3$ showed maximum extractability while $1.0M\;MgSO_4$ solution showed the least extractability. Resonable precipitation was obtained by 40% of acetone and ammonium sulfate. It has been shown by SDS polyacrylamide gel electrophoresis that the soluble protein had 11 bands. The molecular weight for the main protein of the soluble protein wasestimated to be 43,000. In amino acid composition of water extracted protein, arginine content was the highest 47.17% while on the contray, proline and cystine contents were very low.

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Plant Biotechnology and Bioinformatics (식물 생명공학과 생물정보학)

  • Kim, Jung-Eun;Paik, Hyo-Jung;Kim, Young-Cheol;Hur, Cheol-Goo
    • Journal of Plant Biotechnology
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    • v.33 no.3
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    • pp.209-222
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    • 2006
  • The whole genome sequence was completed in arabidopsis and rice. Large amounts of EST data have been available from many other plants. Also, vast quantities of diverse biological data have been generated by various '-omics' technologies such as transcriptomics, proteomics, and metabolomics. Bioinformatics plays an essential role in extracting useful information from these tremendous amounts of biological data. In this review we introduced experimental methods to generate massive data, applications to plant science such as plant disease resistance and molecular breeding and bioinformatics tools and web sites available in plant biotechnology R&D. We concluded that new experimental methods and bioinfomation analysis techniques have made major contributions to the development of plant biotechnology and that bioinformatics has become a critical factor in plant biotechnology R&D.

Copper Oxide Spike Grids for Enhanced Solution Transfer in Cryogenic Electron Microscopy

  • Dukwon Lee;Hansol, Lee;Jinwook Lee;Soung-Hun Roh;Nam-Chul Ha
    • Molecules and Cells
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    • v.46 no.9
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    • pp.538-544
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    • 2023
  • The formation of uniform vitreous ice is a crucial step in the preparation of samples for cryogenic electron microscopy (cryo-EM). Despite the rapid technological progress in EM, controlling the thickness of vitreous ice on sample grids with reproducibility remains a major obstacle to obtaining high-quality data in cryo-EM imaging. The commonly employed classical blotting process faces the problem of excess water that cannot be absorbed by the filter paper, resulting in the formation of thick and heterogeneous ice. In this study, we propose a novel approach that combines the recently developed nanowire self-wicking technique with the classical blotting method to effectively control the thickness and homogeneity of vitrified ice. With simple procedures, we generated a copper oxide spike (COS) grid by inducing COSs on commercially available copper grids, which can effectively remove excess water during the blotting procedure without damaging the holey carbon membrane. The ice thickness could be controlled with good reproducibility compared to non-oxidized grids. Incorporated into other EM techniques, our new modification method is an effective option for obtaining high-quality data during cryo-EM imaging.

Toward High-Resolution Cryo-Electron Microscopy: Technical Review on Microcrystal-Electron Diffraction

  • Lee, Sangmin;Chung, Jeong Min;Jung, Hyun Suk
    • Applied Microscopy
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    • v.47 no.4
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    • pp.223-225
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    • 2017
  • Cryo-electron microscopy (cryo-EM) is arguably the most powerful tool used in structural biology. It is an important analytical technique that is used for gaining insight into the functional and molecular mechanisms of biomolecules involved in several physiological processes. Cryo-EM can be separated into the following three groups according to the analytical purposes and the features of the biological samples: cryo-electron tomography (cryo-ET), cryo-single-particle reconstruction, and cryo-electron crystallography. Cryo-tomography is a unique EM technique that is used to study intact biomolecular complexes within their original environments; it can provide mechanistic insights that are challenging for other EM-methods. However, the resolution of reconstructed three-dimensional (3D) models generated by cryo-ET is relatively low, while single-particle reconstruction can reproduce biomolecular structures having near-atomic resolution without the need for crystallization unless the samples are large (>200 kDa) and highly symmetrical. Cryo-electron crystallography is subdivided into the following two categories according to the types of samples: one category that deals with two-dimensional (2D) crystalline arrays and the other category that uses 3D crystals. These two categories of electron-crystallographic techniques use different diffraction data obtained from still diffraction and continuous-rotation diffraction. In this paper, we review crystal-based cryo-EM techniques and focus on the recently developed 3D electron-crystallographic technique called microcrystal-electron diffraction.

Visualizing Live Chromatin Dynamics through CRISPR-Based Imaging Techniques

  • Chaudhary, Narendra;Im, Jae-Kyeong;Nho, Si-Hyeong;Kim, Hajin
    • Molecules and Cells
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    • v.44 no.9
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    • pp.627-636
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    • 2021
  • The three-dimensional organization of chromatin and its time-dependent changes greatly affect virtually every cellular function, especially DNA replication, genome maintenance, transcription regulation, and cell differentiation. Sequencing-based techniques such as ChIP-seq, ATAC-seq, and Hi-C provide abundant information on how genomic elements are coupled with regulatory proteins and functionally organized into hierarchical domains through their interactions. However, visualizing the time-dependent changes of such organization in individual cells remains challenging. Recent developments of CRISPR systems for site-specific fluorescent labeling of genomic loci have provided promising strategies for visualizing chromatin dynamics in live cells. However, there are several limiting factors, including background signals, off-target binding of CRISPR, and rapid photobleaching of the fluorophores, requiring a large number of target-bound CRISPR complexes to reliably distinguish the target-specific foci from the background. Various modifications have been engineered into the CRISPR system to enhance the signal-to-background ratio and signal longevity to detect target foci more reliably and efficiently, and to reduce the required target size. In this review, we comprehensively compare the performances of recently developed CRISPR designs for improved visualization of genomic loci in terms of the reliability of target detection, the ability to detect small repeat loci, and the allowed time of live tracking. Longer observation of genomic loci allows the detailed identification of the dynamic characteristics of chromatin. The diffusion properties of chromatin found in recent studies are reviewed, which provide suggestions for the underlying biological processes.

Enterotoxigenic Bacteroides fragilis-Associated Diseases and Detection (Enterotoxigenic Bacteroides fragilis에 의한 질환과 검출)

  • Gwon, Sun-Yeong;Jang, In-Ho;Rhee, Ki-Jong
    • Korean Journal of Clinical Laboratory Science
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    • v.47 no.4
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    • pp.161-167
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    • 2015
  • These commensal intestinal bacteria can enhance the immune system and aid in nutrient absorption but can also act as opportunistic pathogens. Among these intestinal bacteria, the anaerobic Bacteroides fragilis are divided into enterotoxigenic B. fragilis (ETBF) which secrete the B. fragilis toxin (BFT) and non-enterotoxigenic B. fragilis (NTBF) which do not secrete BFT. ETBF can cause diarrhea and colitis in both humans and livestock but can also be found in asymptomatic individuals. ETBF is predominantly found in patients with inflammatory diarrheal diseases and traveller's diarrhea. Several clinical studies have also reported an increased prevalence of ETBF in human patients with inflammatory bowel disease (IBD), colitis and colorectal cancer. In small animal models (C57BL/6 wild-type mice, germ-free mice, multiple intestinal neoplasia (Min) mice, rabbits and Mongolian gerbils), ETBF have been found to initiate and/or aggravate IBD, colitis and colorectal cancer. BFT induces E-cadherin cleavage in intestinal epithelial cells resulting in loss of epithelial cell integrity. Subsequent activation of the ${\beta}$-catenin pathway leads to increased cellular proliferation. In addition, ETBF causes acute and chronic colitis in wild-type mice as well as enhances tumorigenesis in Min mice via activation of the Stat3/Th17 pathway. Currently, ETBF can be detected using a BFT toxin bioassay and by PCR. Advances in molecular biological techniques such as real-time PCR have allowed both researchers as well as clinicians to rapidly detect ETBF in clinical samples. The emergence of more sensitive techniques will likely advance molecular insight into the role of ETBF in colitis and cancer.

Layered Double Hydroxide Nanoparticles for Bio-Imaging Applications (LDH 나노입자 기반의 바이오 이미징 소재)

  • Jin, Wenji;Ha, Seongjin;Lee, Dongki;Park, Dae-Hwan
    • Korean Chemical Engineering Research
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    • v.57 no.4
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    • pp.445-454
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    • 2019
  • Layered double hydroxides (LDHs) nanoparticles have emerged as novel nanomaterials for bio-imaging applications due to its unique layered structure, physicochemical properties, and good biocompatibility. Bio-imaging is one of the most important fields for medical applications in clinical diagnostics and therapeutics of various diseases. Enhanced diagnostic techniques are needed to realize new paradigm for next-generation personalized medicine through nanoscale materials. When nanotechnology is introduced into bio-imaging system, nanoparticle probes can endow imaging techniques with enhanced ability to obtain information about biological system at the molecular level. In this review, we summarize structural features of LDH nanoparticles with current issues of bio-imaging system. LDH nanoparticle probes are also discussed through in vitro as well as in vivo studies in various bio-imaging techniques including fluorescence imaging, magnetic resonance imaging (MRI), positron emission tomography (PET), and computed X-ray tomography (CT), which will have the potential in the development of the advanced nanoparticles with high sensitivity and selectivity.