• Title/Summary/Keyword: specific labeling

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The Distribution of ATPase and Porin in the Bovine Heart Mitochondrial Cristae (소(牛) 심근 미토콘드리아의 ATPase와 porin의 분포)

  • Kim, Tae-Keun;Min, Byoung-Hoon;Kim, Soo-Jin
    • Applied Microscopy
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    • v.40 no.4
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    • pp.261-266
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    • 2010
  • ATP is the energy source synthesized at the electron transferase that consist of complex I, II, III, IV and V in mitochondrial cristae. The complex V functions as ATPase which composed of sub-complex $F_0$ and $F_1$. Porin or VDAC (voltagedependent anion-selective channel), is a family of small pore-forming proteins of the mitochondrial outer membrane, and play important roles in the regulated flux of anion, proton and metabolites between the cytosolic and mitochondrial compartments. The channel allows the diffusion of negatively charged solutes such as succinate, malate, and ATP in the fully open state, but of positively charged ions in subconducting state. In this study, in order to investigate the relationship of the function and localization between porin and ATPase we observed the distribution of porin and ATPase in the mitochondria of the bovine heart. Monoclonal antibodies against porin and ATPase ${\beta}$-subunit were used to detect porin and ATPase using light microscope with immunohistochemistry and immunofluorescence, and using electron microscope with immunogold-labeling. ATPase were stained in longitudinal section region in cardiac muscle, porin were stained in longitudinal section region in cardiac muscle. We viewed more specific pattern of localization and distribution of these proteins using immunofluorescence method. There were some region which were labeled with porin or ATPase respectively, and others which were labeled both proteins in cardiac muscle. The electron microscope results showed that immunogold labeled porin were labeled locally at mitochondrial outer membrane and ATPase were labeled evenly at mitochondrial cristae. But ATPase was not labeled at mitochondria cristae. These results confirmed the subcellular localizations of porin and ATPase in mitochondrial outer membrane and cristae. Also, we assumed that ATP synthesis always does not activation in all mitochondria exist in the bovine cardiac muscle.

Development of PCR Method for Rapid Detection of Allergic Materials in Foods (PCR을 이용한 식품 중 알레르기 유발물질 검출법 개발)

  • Park, Yong-Chjun;Kim, Mi-Ra;Shin, Jun-Ho;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Lee, Sang-Jae;Han, Sang-Bae
    • Journal of Food Hygiene and Safety
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    • v.28 no.2
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    • pp.124-129
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    • 2013
  • The method for detection foods containing allergenic materials by PCR was developed in this study. To detect allergenic raw material from processed food, species specific primer which up to 200bp for PCR product were designed or selected from advanced research. As target materials, 14 items were selected (12 target materials for allergen in Korea, 2 target materials for allergen in foreign countries). The amplicon size for eggs, milk, buckwheat, peanuts, beans, wheat, mackerel, crab, shrimp, pork, peach, tomato, almond, and sesame were confirmed 281, 131, 138, 120, 118, 127, 211, 174, 231, 138, 174, 132, 103, and 220bp, respectively. And any non-specific bands were not detected among each others. Detection method for allergenic material developed in this study could be used to investigate inaccurate goods for allergen labeling or non-intentional contaminant during processed foods manufacturing. In addition, the system will be usefully to detection accurate allergenic raw materials of export for other countries.

The Production of Sex Determined Cattle by Embryonic Sexing Using Fluorescence In Situ Hybridization Technique (FISH 기법을 이용한 소 수정란의 성감별과 산자 생산)

  • Sohn, S.H.;Park, H.
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • 2007.05a
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    • pp.39-50
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    • 2007
  • Sexing from bovine embryos fertilized in vitro implicates a possibility of the sex controlled cattle production. This study was carried out to produce the sex determined cattle through the embryonic sexing by fluorescence in situ hybridization (FISH) technique. FISH was achieved in in vitro fertilized bovine embryos using a bovine Y-specific DNA probe constructed from the btDYZ-1 sequence. Using this probe, a male-specific signal was detected on 100% of Y-chromosome bearing metaphase specimens. The analyzable rate of embryonic sexing by FISH technique was about 93% (365/393) regardless of embryonic stages. As tested single blastomere by FISH and then karyotype with their biopsied embryos, the accuracy of sex determination with FISH was 97.6%. We tried the embryo transfer with sex determined embryos on 15 cattle. Among them, the 5 cattle delivered calf with expected sex last year.

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Monitoring of Restaurant Beef Labeling System (음식점 식육 원산지 표시 모니터링)

  • Hong, Jin;Leem, Dong-Gil;Kim, Mi-Gyeong;Park, Kyoung-Sik;Yoon, Tae-Hyung;No, Ki-Mi;Jeong, Ja-Young
    • Journal of Food Hygiene and Safety
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    • v.25 no.2
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    • pp.162-169
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    • 2010
  • The compulsory beef labelling system has launched from January 1st 2007 by the amended Food Hygiene Law, we were checked the actual conditions of beef origin with a nationwide scale by the Hanwoo differentiation specific test method which was developed by Korea FDA using 90 SNP biomarkers. The test method is useful tool to differentiate the beef origin carrying out the mission of KFDA's annual food safety management guidance. Also we have technically transferred the Hanwoo differentiation specific test method to other institutes as well regional KFDA and established the training program as a regular course in Korea Human Resource Development Institute for Health and Welfare. The beef used in this study were collected according to the 2009 Food safety guidance in roast beef restaurants where business site area greater than 100 $m^2$. Total 216 samples were consisted of 48 samples of the Seoul area and 168 of the region. The monitoring result from restaurants in all the region of Korea showed that 3 of 216 Hanwoo-labelled beefs were found out as a non-Hanwoo (1.3%). This results are gradually deceasing trend compared with 34.0% in 2005, 30.1% in 2006, 3.2% in 2007 and 5.14% in 2008. From these data, the Hanwoo differentiation specific test method on the settlement of the compulsory beef labelling system has an important role. As a outcome of this project, we might be considered the early settlement of the compulsory beef labelling system, technically transferred to other institutes and the establishment of regular training program of the test method.

High Glucose Causes Human Cardiac Progenitor Cell Dysfunction by Promoting Mitochondrial Fission: Role of a GLUT1 Blocker

  • Choi, He Yun;Park, Ji Hye;Jang, Woong Bi;Ji, Seung Taek;Jung, Seok Yun;Kim, Da Yeon;Kang, Songhwa;Kim, Yeon Ju;Yun, Jisoo;Kim, Jae Ho;Baek, Sang Hong;Kwon, Sang-Mo
    • Biomolecules & Therapeutics
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    • v.24 no.4
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    • pp.363-370
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    • 2016
  • Cardiovascular disease is the most common cause of death in diabetic patients. Hyperglycemia is the primary characteristic of diabetes and is associated with many complications. The role of hyperglycemia in the dysfunction of human cardiac progenitor cells that can regenerate damaged cardiac tissue has been investigated, but the exact mechanism underlying this association is not clear. Thus, we examined whether hyperglycemia could regulate mitochondrial dynamics and lead to cardiac progenitor cell dysfunction, and whether blocking glucose uptake could rescue this dysfunction. High glucose in cardiac progenitor cells results in reduced cell viability and decreased expression of cell cycle-related molecules, including CDK2 and cyclin E. A tube formation assay revealed that hyperglycemia led to a significant decrease in the tube-forming ability of cardiac progenitor cells. Fluorescent labeling of cardiac progenitor cell mitochondria revealed that hyperglycemia alters mitochondrial dynamics and increases expression of fission-related proteins, including Fis1 and Drp1. Moreover, we showed that specific blockage of GLUT1 improved cell viability, tube formation, and regulation of mitochondrial dynamics in cardiac progenitor cells. To our knowledge, this study is the first to demonstrate that high glucose leads to cardiac progenitor cell dysfunction through an increase in mitochondrial fission, and that a GLUT1 blocker can rescue cardiac progenitor cell dysfunction and downregulation of mitochondrial fission. Combined therapy with cardiac progenitor cells and a GLUT1 blocker may provide a novel strategy for cardiac progenitor cell therapy in cardiovascular disease patients with diabetes.

Immunochemical Studies for the Characterization of Purified $(Na^+,\;K^+)-ATPase$ and Its Subunits with a Special Reference of Their Effect on Monovalent Cation Transport in Reconstituted $(Na^+,\;K^+)-ATPase$ Vesicles

  • Rhee, H.M.;Hokin, L.E.
    • The Korean Journal of Pharmacology
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    • v.26 no.1
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    • pp.35-49
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    • 1990
  • A highly purified $(Na^+,\;K^+)-ATPase$ from the rectal gland of Squalus acanthias and from the electric organ of Electrophorus electricus has been used to raise antibodies in rabbits. The 97,000 dalton catalytic subunit and glycoprotein derived from the rectal gland of spiny shark were also used as antigens. The two $(Na^+,\;K^+)-ATPase$ holoenzymes and the two shark subunits were antigenic. In Ouchterlony double diffusion experiments, these antibodies formed precipitation bands with their antigens. Antibodies prepared against the two subunits of shark holoenzyme also formed precipitation bands with their antigens and shark holoenzyme, but not with eel holoenzyme. These observations are in good agreement with inhibitory effect of these antibodies on the catalytic activity of $(Na^+,\;K^+)-ATPase$ both from the shark and the eel, since there is very little cross-reaction between the shark anticatalytic subunit antibodies and the eel holoenzyme. The maximum antibodies titer of the anticatalytic subunit antibodies is found to be 6 weeks after the initial single exposure to this antigen. Multiple injections of the antigen increased the antibody titer. However, the time required to produce the maximum antibody titer was approximately the same. These antibodies also inhibit catalytic activity of $(Na^+,\;K^+)-ATPase$ vesicles reconstituted by a slow dialysis of cholate after solubilization of the enzyme in a presonicated mixture of cholate and phospholipid. In these reconstituted $(Na^+,\;K^+)-ATPase$ vesicles, effects of these antibodies on the fluxes of $Na^+$, $Rb^+$, and $K^+$ were investigated. Control or preimmune serum had no effect on the influx of $^{22}Na^+$ or the efflux of $^{86}Rb^+$. Immunized sera against the shark $(Na^+,\;K^+)-ATPase$ holoenzyme, its glycoprotein or catalytic subunit did inhibit the influx of $^{22}Na^+$ and the efflux of $^{86}Rb^+$. It was also demonstrated that these antibodies inhibit the coupled counter-transport of $Na^+$ and $K^+$ as studied by means of dual labeling experiments. However, this inhibitory effect of the antibodies on transport of ions in the $(Na^+,\;K^+)-ATPase$ vesicles is manifested only on the portion of energy and temperature dependent alkali metal fluxes, not on the portion of ATP and ouabain insensitive ion movement. Simultaneous determination of effects of the antibodies on ion fluxes and vesicular catalytic activity indicates that an inhibition of active ion transport in reconstituted $(Na^+,\;K^+)-ATPase$ vesicles appears to be due to the inhibitory action of the antibodies on the enzymatic activity of $(Na^+,\;K^+)-ATPase$ molecules incorporated in the vesicles. These findings that the inhibitory effects of the antibodies specific to $(Na^+,\;K^+)-ATPase$ or to its subunits on ATP and temperature sensitive monovalent cation transport in parallel with the inhibitory effect of vesicular catalytic activity by these antibodies provide direct evidence that $(Na^+,\;K^+)-ATPase$ is the molecular machinery of active cation transport in this reconstituted $(Na^+,\;K^+)-ATPase$ vesicular system.

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Synaptic Pattern of NMDA R1 upon the Direction-Selective Retinal Ganglion Cells in Developing Mouse Retina (발생 중 마우스 망막에서 방향특이성 신경절세포의 NMDA R1 수용체의 시냅스 패턴)

  • Lee, Jee-Geon;Kwon, Oh-Ju;Jeon, Chang-Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.18 no.4
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    • pp.533-540
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    • 2013
  • Purpose: To investigate the synaptic pattern of NMDA glutamate receptor subtype NMDA R1 on the dendritic arbors of ON-OFF direction-selective retinal ganglion cells (DS-RGSs) in developing [(5,10) days postnatal (PN)] mouse retina. Methods: ON-OFF DS-RGCs were injected with Lucifer yellow and the cells were identified by their characteristic morphology. To identify glutamatergic excitatory input from bipolar cell, we used a marker for the membrane traffic motor protein kinesin. Results: We identified DS-RGCs in P5, and P10 mouse retina. The immunofluorescence labeling of NMDA R1 was most prominent in the IPL. Our results showed that their presence upon the entire dendritic arbor of ON-OFF DS-RGCs is without any evidence of asymmetry, which would predict direction selectivity. Conclusions: The glutamatergic input from bipolar cell reveals symmetry pattern in all periods of P5, and P10. The results may suggest that direction selectivity not lies in the specific pattern of NMDA R1 receptors.

Integration of Component Image Information and Design Information by Graph to Support Product Design Information Reuse (제품 설계 정보 재사용을 위한 그래프 기반의 부품 영상 정보와 설계 정보의 병합)

  • Lee, Hyung-Jae;Yang, Hyung-Jeong;Kim, Kyoung-Yun;Kim, Soo-Hyung;Kim, Sun-Hee
    • The KIPS Transactions:PartD
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    • v.13D no.7 s.110
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    • pp.1017-1026
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    • 2006
  • Recently, distributed collaborative development environment has been recognized an alternative environment for product development in which multidisciplinary participants are naturally involving. Reuse of Product design information has long been recognized as one of core requirements for efficient product development. This paper addresses an image-based retrieval system to support product design information reuse. In the system, product images obtained from multi-modal devices are utilized to reuse design information. The proposed system conducts the segmentation of a product image by using a labeling method and generates an attributed relational graph (ARG) that represents properties of segmented regions and their relationships. The generated ARG is extended by integrating corresponding part/assembly information. In this manner, the reuse of assembly design information using a product image has been realized. The main advantages of the presented system are following. First, the system is not dependent to specific design tools, because it utilizes multimedia images that can be obtained easily from peripheral devices. Second ratio-based features extracted from images enable image retrievals that contain various sizes of parts. Third, the system has shown outstanding search performance, because we applied various information of segmented part regions and their relationships between parts.

Biodistribution of 99mTc Labeled Integrin Antagonist

  • Jang, Beom-Su;Park, Seung-Hee;Shin, In Soo;Maeng, Jin-Soo;Paik, Chang H.
    • Toxicological Research
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    • v.29 no.1
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    • pp.21-25
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    • 2013
  • The selective targeting of an integrin ${\alpha}_v{\beta}_3$ receptor using radioligands may enable the assessment of angiogenesis and integrin ${\alpha}_v{\beta}_3$ receptor status in tumors. The aim of this research was to label a peptidomimetic integrin ${\alpha}_v{\beta}_3$ antagonist (PIA) with $^{99m}Tc(CO)_3$ and to test its receptor targeting properties in nude mice bearing receptor-positive tumors. PIA was reacted with tris-succinimidyl aminotriacetate (TSAT) (20 mM) as a PIA per TSAT. The product, PIA-aminodiacetic acid (ADA), was radiolabeled with $[^{99m}Tc(CO)_3(H_2O)_3]^{+1}$, and purified sequentially on a Sep-Pak C-18 cartridge followed by a Sep-Pak QMA anion exchange cartridge. Using gradient C-18 reverse-phase HPLC, the radiochemical purity of $^{99m}Tc(CO)_3$-ADA-PIA (retention time, 10.5 min) was confirmed to be > 95%. Biodistribution analysis was performed in nude mice (n = 5 per time point) bearing receptor-positive M21 human melanoma xenografts. The mice were administered $^{99m}Tc(CO)_3$-ADA-PIA intravenously. The animals were euthanized at 0.33, 1, and 2 hr after injection for the biodistribution study. A separate group of mice were also co-injected with 200 ${\mu}g$ of PIA and euthanized at 1 hr to quantify tumor uptake. $^{99m}Tc(CO)_3$-ADA-PIA was stable in phosphate buffer for 21 hr, but at 3 and 6 hr, 7.9 and 11.5% of the radioactivity was lost as histidine, respectively. In tumor bearing mice, $^{99m}Tc(CO)_3$-ADA-PIA accumulated rapidly in a receptor-positive tumor with a peak uptake at 20 min, and rapid clearance from blood occurring primarily through the hepatobiliary system. At 20 min, the tumor-to-blood ratio was 1.8. At 1 hr, the tumor uptake was 0.47% injected dose (ID)/g, but decreased to 0.12% ID/g when co-injected with an excess amount of PIA, indicating that accumulation was receptor mediated. These results demonstrate successful $^{99m}TC$ labeling of a peptidomimetic integrin antagonist that accumulated in a tumor via receptor-specific binding. However, tumor uptake was very low because of low blood concentrations that likely resulted from rapid uptake of the agent into the hepatobiliary system. This study suggests that for $^{99m}Tc(CO)_3$-ADA-PIA to be useful as a tumor detection agent, it will be necessary to improve receptor binding affinity and increase the hydrophilicity of the product to minimize rapid hepatobiliary uptake.

Developing a Korean Standard Brain Atlas on the basis of Statistical and Probabilistic Approach and Visualization tool for Functional image analysis (확률 및 통계적 개념에 근거한 한국인 표준 뇌 지도 작성 및 기능 영상 분석을 위한 가시화 방법에 관한 연구)

  • Koo, B.B.;Lee, J.M.;Kim, J.S.;Lee, J.S.;Kim, I.Y.;Kim, J.J.;Lee, D.S.;Kwon, J.S.;Kim, S.I.
    • The Korean Journal of Nuclear Medicine
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    • v.37 no.3
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    • pp.162-170
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    • 2003
  • The probabilistic anatomical maps are used to localize the functional neuro-images and morphological variability. The quantitative indicator is very important to inquire the anatomical position of an activated legion because functional image data has the low-resolution nature and no inherent anatomical information. Although previously developed MNI probabilistic anatomical map was enough to localize the data, it was not suitable for the Korean brains because of the morphological difference between Occidental and Oriental. In this study, we develop a probabilistic anatomical map for Korean normal brain. Normal 75 blains of T1-weighted spoiled gradient echo magnetic resonance images were acquired on a 1.5-T GESIGNA scanner. Then, a standard brain is selected in the group through a clinician searches a brain of the average property in the Talairach coordinate system. With the standard brain, an anatomist delineates 89 regions of interest (ROI) parcellating cortical and subcortical areas. The parcellated ROIs of the standard are warped and overlapped into each brain by maximizing intensity similarity. And every brain is automatically labeledwith the registered ROIs. Each of the same-labeled region is linearly normalize to the standard brain, and the occurrence of each legion is counted. Finally, 89 probabilistic ROI volumes are generated. This paper presents a probabilistic anatomical map for localizing the functional and structural analysis of Korean normal brain. In the future, we'll develop the group specific probabilistic anatomical maps of OCD and schizophrenia disease.