• Title/Summary/Keyword: surface localization

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A PAS-Containing Histidine Kinase is Required for Conidiation, Appressorium Formation, and Disease Development in the Rice Blast Fungus, Magnaporthe oryzae

  • Shin, Jong-Hwan;Gumilang, Adiyantara;Kim, Moon-Jong;Han, Joon-Hee;Kim, Kyoung Su
    • Mycobiology
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    • v.47 no.4
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    • pp.473-482
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    • 2019
  • Rice blast disease, caused by the ascomycete fungus Magnaporthe oryzae, is one of the most important diseases in rice production. PAS (period circadian protein, aryl hydrocarbon receptor nuclear translocator protein, single-minded protein) domains are known to be involved in signal transduction pathways, but their functional roles have not been well studied in fungi. In this study, targeted gene deletion was carried out to investigate the functional roles of the PAS-containing gene MoPAS1 (MGG_02665) in M. oryzae. The deletion mutant ΔMopas1 exhibited easily wettable mycelia, reduced conidiation, and defects in appressorium formation and disease development compared to the wild type and complemented transformant. Exogenous cAMP restored appressorium formation in ΔMopas1, but the shape of the restored appressorium was irregular, indicating that MoPAS1 is involved in sensing the hydrophobic surface. To examine the expression and localization of MoPAS1 in M. oryzae during appressorium development and plant infection, we constructed a MoPAS1:GFP fusion construct. MoPAS1:GFP was observed in conidia and germ tubes at 0 and 2 h post-infection (hpi) on hydrophobic cover slips. By 8 hpi, most of the GFP signal was observed in the appressoria. During invasive growth in host cells, MoPAS1:GFP was found to be fully expressed in not only the appressoria but also invasive hyphae, suggesting that MoPAS may contribute to disease development in host cells. These results expand our knowledge of the roles of PAS-containing regulatory genes in the plant-pathogenic fungus M. oryzae.

Toxoplasma gondii: Ultrastructural localization of specific antigens and inhibition of intracellular multiplication by monoclonal antibodies

  • Lee, Boo-Young;Ahn, Myoung-Hee;Kim, Hyun-Chul;Min, Duk-Young
    • Parasites, Hosts and Diseases
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    • v.39 no.1
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    • pp.67-76
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    • 2001
  • This experiment was focused on the characterization of anti- Toxoplasma monoclonal antibodies (mAbs) and the effect of mAbs on the parasite invasion of mouse peritoneal macrophages. Twenty eight mAbs including M110, M556, R7A6 and M62l were characterized by Ab titer, immunoglobulin isotyping and western blot pattern. Antibody titer (optical density) of 4 mAbs. Ml 10. M556. R7A6 and M62l. were 0.53,0.67, 0.45 and 0.39 (normal mouse serum; 0.19) with the same IgGl isotypes shown by Enzyme-linked immunosorbent assay (ELISA). Western blot analysis showed that Ml 10. M556. R7A6 and M62l reacted with the 33 kDa (p30),31 kDa (p28),43 kDa and 36 kDa protein. Immuno-gold labelling of mAbs M110, M556, R7A6 and M621 reacted with the surface membrane, dense granules and parasitophorous vacuolar membrane (PVM) , rhoptries and cytoplasm of tachyzoite, respectively. For in vitro assay, preincubation of tachyzoties with four mAbs, Ml 10, M556, R7A6 and M62l resulted in the decrease of the number of infected macrophages (p < 0.05) and the suppression of parasite multiplication at 18 h post-infection. Four monoclonal antibodies including Ml 10 (SAGI) were found to have an important role in the inhibition of macrophage invasion and T. gondii multiplication in vitro, and these mAbs may be suitable for vaccine candidates, diagnostic kit and for chemotherapy.

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Loci of Orebodies, the Bupyeong Silver Deposits (부평은광상(富平銀鑛床)의 광체배태장소(鑛體胚胎場所))

  • Suh, Kyu-Sik;Park, Hee-In
    • Economic and Environmental Geology
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    • v.20 no.2
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    • pp.97-106
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    • 1987
  • The geology of the Bupyeong mine area is consisted of Precambrian Gyeonggi gneiss complex and Mesozoic igneous rocks; i.e., pyroclastic rocks, intrusive breccia, granite and felsic porphyries which were formed during a Jurassic to early Cretaceous resurgent caldera evolution. Granites are not observed on the surface and in the underground of the mine. Bupyeong silver deposits occur as stockworks of base metal sulfides- minor silver minerals-quartz - carbonate veinlets, hosted by pyroclastic rocks and intrusive breccia at the southwestern margin of the caldera. Silver occurs mainly as native silver, and other silver minerals, minor in quantity, are argentite, tetrahedrite-freibergite, pyrargyrite, polybasite, canfieldite and dyscrasite. The average grade of silver ore is about 180g/t Ag. Discrimination of silver ore from the country rocks depends largely on the chemical analyses of rock samples taken every two meters from tunnels, diamond-drilling cores and mining stopes, because silver minerals are hardly observed in the ore by crude eye, and silver orebodies do not properly coincide with the concentrated zone of base metal sulfides which were precipitated at the earlier stage than the stage of precipitation of native silver. General characteristics of the loci of the silver orebodies are as follows; (1) The host rocks of orebodies are pyroclastic rocks and intrusive breccia. (2) Many of the orebodies are distributed around Gyeonggi gneiss complex. Especially where the paleotopography of gneiss complex shows a gradual slope, the basal stratigraphic horizon of the pyroclastic rocks unconformably overlying the gneiss complex offered a favorable loci of high grade ore. (3) $N5^{\circ}W$ to $N15^{\circ}$ E-striking faults played an important role in the localization of the orebodies. (4) Conduits of intrusive breccia within the gneiss complex, through which the intrusive breccia intruded into the upper pyroclastic rocks, exist beneath most of the main orebodies. This suggests that the conduits of intrusive breccia served as channelways for the migration of ore fluids.

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Construction and Operation of a 37-channel Hemispherical Magnetoencephalogram System (37채널 반구형 뇌자도 측정장치 제작 및 동작)

  • 이용호;김진목;권혁찬;김기웅;박용기;강찬석;이순걸
    • Journal of Biomedical Engineering Research
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    • v.24 no.3
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    • pp.159-165
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    • 2003
  • We developed a 37-channel magnetoencephalogram (MEG) measurement system based on low-noise superconducting quantum interference device (SQUID) magnetometets, and operated the system to measure MEG signals. By using double relaxation oscillation SQUIDs with high flux-4o-voltage transfers, the SQUID outputs could be measured directly by room temperature preamplifiers and compact readout circuits were used for SQUID operation. The average field noise level of the magnetometers is about 3 fT/√Hz in the white region, low enough for MEG measurements when operated inside a magnetically shielded room. The 37 magnetometers were distributed on a hemispherical surface haying a radius of 125 mm. In addition to the 37 sensing channels. 11 reference channels were installed to pickup external noise and to form software gradiometers. A low-noise liquid helium dewar was fabricated with a liquid capacity of 30 L and boil-off rate of 4 L/d. The signal processing software consists of digital filtering, software gradiometer, isofield mapping and source localization. By using the developed system, we measured auditory-evoked fields and localized the current dipoles, demonstrating the effectiveness of the system.

Distribution of Lipid and Lipase in Lipid-and Starch-Rich Seeds (지질 및 전분성 종자에서 지질 및 지질가수분해효소의 분포)

  • 김우갑
    • Journal of Plant Biology
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    • v.35 no.3
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    • pp.219-227
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    • 1992
  • Formation, cellular distribution and structural changes of storage lipid, and active site and cellular localization of lipase in endosperms and cotyledons of lipid-rich seeds such as Helianthus annuus, Ricinus communis and Pinus koraiensis, and in those of starch-rich seeds such as Pisum sativum and Zea mays were investigated in relation to the seed development by cytochemical methods. In endosperms and storage cotyledons of lipid- and starch-rich seeds after seed-gathering, there were widely distributed storage material which was composed of spherical protein bodies, spherosomes, and starch granules. But cellular organelles were hardly observed in the cytoplasm. Staining pattern of vesicles released from SER, and of low electron dense membraneous granules, which were perhaps at an early stage of spherosomes, were the same as in the spherosome. Electrondense granules released from RER were observed in the vicinity of plasma membrane. As a result of lipid staining, the spherosomes were more electron dense and were uniform as compared with the protein matrix within the protein body and cytoplasmic proteinaceous granules. The major component of the spherosome was determinated to be lipid. Spherosomes and vesicles containing SER-released materials showed the same as in the electron density. Lipase activity was especially strong in the inner region and on the surface of decomposed spherosomes and near the plasma membrane.mbrane.

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Membrane and Virus Filter Trends in the Processes of Biopharmaceutical Production (바이오의약품 제조공정에서 분리막의 역할과 바이러스 필터 동향)

  • Choi, Tae Hwan;Park, Ho Bum
    • Membrane Journal
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    • v.30 no.1
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    • pp.9-20
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    • 2020
  • Membranes are used in most processes of biopharmaceutical production. It is used for pretreatment of other processes, separation of impurities in the process, virus removal, control of products concentration and buffer solution exchange. Virus filters play an important role in ensuring product efficacy and stability because viral contamination of biopharmaceuticals for humans is a sensitive issue that is directly related to serious clinical outcomes. Virus filters typically have complex multilayer structures made of various polymers such as surface-modified PVDF, PES, CRC. Depending on the manufacturer, filters have different pore structures and shapes, such as symmetric or asymmetric, and is used in the form of pleated membrane, flat sheets or hollow fibers. Virus filters are exclusively supplied by few foreign companies such as Asahi Kasei, Millipore, Pall and Sartorius. Replacing virus filters can be time consuming and expensive, including approval from regulatory agencies through validation. As localization has become important due to Japan's recent export regulations, it is necessary to increase the degree of technical independence.

Cross-linking of CD80 and CD86 Diminishes Expression of CD54 on EBV-transformed B Cells through Inactivation of RhoA and Ras

  • Park, Ga-Bin;Kim, Yeong-Seok;Song, Hyun-Keun;Kim, Seong-Han;Park, Dong-Man;Lee, Wang-Jae;Hur, Dae-Young
    • IMMUNE NETWORK
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    • v.11 no.6
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    • pp.390-398
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    • 2011
  • Background: Epstein Barr virus (EBV) infected B cells are transformed into lymphoblastoid cell lines. Some researchers suggested some a few similarities between this process and carcinogenesis. We observed the expression of CD80 and CD86, co-stimulatory molecules on EBV-transformed B cells and changes of CD54 expression after stimulation of CD80 and CD86. Methods: CD80 and CD86 were stimulated using anti-CD80 and anti-CD86 monoclonal antibodies. To assess apoptosis and surface protein expression, flow cytometric analysis was performed. Intracellular signal molecules were evaluated by RT-PCR and immunoblot. Morphology and localization of proteins were examined using inverted or confocal microscope. Results: Cross-linking of CD80 and CD86 induced apoptosis and interfered with proliferation of EBV-transformed B cells, and dispersion of clumped cells. We also examined that their stimulation induced ROS accumulation and reduced CD54 expression. Interestingly, we observed that CD80 and CD86 diminished the expression of CD54 in different methods. Both CD80 and CD86 downregulated activation of focal adhesion kinase. CD80 stimulus inhibited CD54 expression through mainly RhoA inactivation, while CD86 down-regulated Ras and JNK phosphorylation. Conclusion: These results suggest that co-stimulatory CD80 and CD86 molecules, expressed EBV-transformed B cells, may play a role in apoptosis and cell adhesion.

Differential Expressions of Aquaporin Subtypes in Female Reproductive Tract of Mice

  • Im, Ji Woo;Lee, Chae Young;Kim, Dong-Hwan;Bae, Hae-Rahn
    • Development and Reproduction
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    • v.24 no.3
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    • pp.177-185
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    • 2020
  • Although many aquaporin (AQP) transcripts have been demonstrated to express in the female reproductive tract, the defined localizations and functions of AQP subtype proteins remain unclear. In this study, we investigated the expression of AQP1, AQP3, AQP5, AQP6, and AQP9 proteins in female reproductive tract of mouse and characterized their precise localizations at the cellular and subcellular levels. Immunofluorescence analyses for AQP1, AQP3, AQP6, and AQP9 showed that these proteins were abundantly expressed in female reproductive tract and that intense immunoreactivities were observed in mucosa epithelial cells with a subtype-specific pattern. The most abundant aquaporin in both vagina and uterine cervix was AQP3. Each of AQP1, AQP3, AQP6, and AQP9 exhibited its distinct distribution in stratified squamous or columnar epithelial cells. AQP9 expression was predominant in oviduct and ovary. AQP1, AQP3, AQP6, and AQP9 proteins were mostly seen in apical membrane of ciliated epithelial cells of the oviduct as well as in both granulosa and theca cells of ovarian follicles. Most of AQP subtypes were also expressed in surface epithelial cells and glandular cells of endometrium in the uterus, but their expression levels were relatively lower than those observed in the vagina, uterine cervix, oviduct and ovary. This is the first study to investigate the expression and localization of 5 AQP subtype proteins simultaneously in female reproductive tract of mouse. Our results suggest that AQP subtypes work together to transport water and glycerol efficiently across the mucosa epithelia for lubrication, proliferation, energy metabolism and pH regulation in female reproductive tract.

LAMP-3 (Lysosome-Associated Membrane Protein 3) Promotes the Intracellular Proliferation of Salmonella typhimurium

  • Lee, Eun-Ju;Park, Kwan-Sik;Jeon, In-Sook;Cho, Jae-Woon;Lee, Sang-Jeon;Choy, Hyun E.;Song, Ki-Duk;Lee, Hak-Kyo;Choi, Joong-Kook
    • Molecules and Cells
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    • v.39 no.7
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    • pp.566-572
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    • 2016
  • Lysosomes are cellular organelles containing diverse classes of catabolic enzymes that are implicated in diverse cellular processes including phagocytosis, autophagy, lipid transport, and aging. Lysosome-associated membrane proteins (LAMP-1 and LAMP-2) are major glycoproteins important for maintaining lysosomal integrity, pH, and catabolism. LAMP-1 and LAMP-2 are constitutively expressed in Salmonella-infected cells and are recruited to Salmonella-containing vacuoles (SCVs) as well as Salmonella- induced filaments (Sifs) that promote the survival and proliferation of the Salmonella. LAMP-3, also known as DC-LAMP/CD208, is a member of the LAMP family of proteins, but its role during Salmonella infection remains unclear. DNA microarray analysis identified LAMP-3 as one of the genes responding to LPS stimulation in THP-1 macrophage cells. Subsequent analyses reveal that LPS and Salmonella induced the expression of LAMP-3 at both the transcriptional and translational levels. Confocal Super resolution N-SIM imaging revealed that LAMP-3, like LAMP-2, shifts its localization from the cell surface to alongside Salmonella. Knockdown of LAMP-3 by specific siRNAs decreased the number of Salmonella recovered from the infected cells. Therefore, we conclude that LAMP-3 is induced by Salmonella infection and recruited to the Salmonella pathogen for intracellular proliferation.

Augmented Reality to Localize Individual Organ in Surgical Procedure

  • Lee, Dongheon;Yi, Jin Wook;Hong, Jeeyoung;Chai, Young Jun;Kim, Hee Chan;Kong, Hyoun-Joong
    • Healthcare Informatics Research
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    • v.24 no.4
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    • pp.394-401
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    • 2018
  • Objectives: Augmented reality (AR) technology has become rapidly available and is suitable for various medical applications since it can provide effective visualization of intricate anatomical structures inside the human body. This paper describes the procedure to develop an AR app with Unity3D and Vuforia software development kit and publish it to a smartphone for the localization of critical tissues or organs that cannot be seen easily by the naked eye during surgery. Methods: In this study, Vuforia version 6.5 integrated with the Unity Editor was installed on a desktop computer and configured to develop the Android AR app for the visualization of internal organs. Three-dimensional segmented human organs were extracted from a computerized tomography file using Seg3D software, and overlaid on a target body surface through the developed app with an artificial marker. Results: To aid beginners in using the AR technology for medical applications, a 3D model of the thyroid and surrounding structures was created from a thyroid cancer patient's DICOM file, and was visualized on the neck of a medical training mannequin through the developed AR app. The individual organs, including the thyroid, trachea, carotid artery, jugular vein, and esophagus were localized by the surgeon's Android smartphone. Conclusions: Vuforia software can help even researchers, students, or surgeons who do not possess computer vision expertise to easily develop an AR app in a user-friendly manner and use it to visualize and localize critical internal organs without incision. It could allow AR technology to be extensively utilized for various medical applications.