• Title/Summary/Keyword: Visual cells

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Development and Optimization of a Rapid Colorimetric Membrane Immunoassay for Porphyromonas gingivalis

  • Lee, Jiyon;Choi, Myoung-Kwon;Kim, Jinju;Chun, SeChul;Kim, Hong-Gyum;Lee, HoSung;Kim, JinSoo;Lee, Dongwook;Han, Seung-Hyun;Yoon, Do-Young
    • Journal of Microbiology and Biotechnology
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    • v.31 no.5
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    • pp.705-709
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    • 2021
  • Porphyromonas gingivalis (P. gingivalis) is a major bacterial pathogen that causes periodontitis, a chronic inflammatory disease of tissues around the teeth. Periodontitis is known to be related to other diseases, such as oral cancer, Alzheimer's disease, and rheumatism. Thus, a precise and sensitive test to detect P. gingivalis is necessary for the early diagnosis of periodontitis. The objective of this study was to optimize a rapid visual detection system for P. gingivalis. First, we performed a visual membrane immunoassay using 3,3',5,5'-tetramethylbenzidine (TMB; blue) and coating and detection antibodies that could bind to the host laboratory strain, ATCC 33277. Antibodies against the P. gingivalis surface adhesion molecules RgpB (arginine proteinase) and Kgp (lysine proteinase) were determined to be the most specific coating and detection antibodies, respectively. Using these two selected antibodies, the streptavidin-horseradish peroxidase (HRP) reaction was performed using a nitrocellulose membrane and visualized with a detection range of 103-105 bacterial cells/ml following incubation for 15 min. These selected conditions were applied to test other oral bacteria, and the results showed that P. gingivalis could be detected without cross-reactivity to other bacteria, including Streptococcus mutans and Escherichia fergusonii. Furthermore, three clinical strains of P. gingivalis, KCOM 2880, KCOM 2803, and KCOM 3190, were also recognized using this optimized enzyme immunoassay (EIA) system. To conclude, we established optimized conditions for P. gingivalis detection with specificity, accuracy, and sensitivity. These results could be utilized to manufacture economical and rapid detection kits for P. gingivalis.

Optimal Conditions of Single Cell Gel Electrophoresis (Comet) Assay to detect DNA single strand breaks in Mouse Lymphoma L5178Y cells

  • Ryu, Jae-Chun;Kwon, Oh-Seung;Kim, Hyung-Tae
    • Environmental Mutagens and Carcinogens
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    • v.21 no.2
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    • pp.89-94
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    • 2001
  • Recently, single cell gel electrophoresis, also known as comet assay, is widely used for the detection and measurement of DNA strand breaks in vitro and in vivo in many toxicological fields such as radiation exposure, human monitoring and toxicity evaluation. As well defined, comet assay is a sensitive, rapid and visual method for the detection of DNA strand breaks in individual cells. Briefly, a small number of damaged cells suspended in a thin agarose gel on a microscope slide were lysed, unwinded, electrophoresed, and stained with a fluorescent DNA binding dye. The electric current pulled the charged DNA from the nucleus such that relaxed and broken DNA fragments migrated further. The resulting images which were subsequently named for their appearance as comets, were measured to determine the extent of DNA damages. However, some variations could be occurred in procedures, laboratories's conditions and kind of cells used. Hence, to overcome and to harmonize these matters in comet assay, International Workshop on Genotoxicity Test Procedure (IWGTP) was held with several topics including comet assay at Washington D.C. on March, 1999. In spite of some consensus in procedures and conditions in IWGTP, there are some problems still remained to be solved. In this respect, we attempted to set the practical optimal conditions in the experimental procedures such as lysis, unwinding, electrophoresis and neutralization conditions and so on. First of all, we determined optimal lysis and unwinding time by using 150 $\mu$M methyl methanesulfonate (MMS) which is usually used concentration. And then, we determined optimal positive control concentrations of benzo(a)pyrene (BaP) and MMS in the presence and absence of S9 metabolic activation system, respectively.

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Histochemical Analysis of the Cone cells in the Retina of the Greater Horseshoe Bat Rhinolophus ferrumequinum (한국관박쥐 망막 원뿔세포의 조직화학적 분석)

  • Jeon, Young-Ki;Joo, Young-Rak;Ye, Eun-Ah;Kim, Moon-Sook;Jeon, Chang-Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.18 no.2
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    • pp.187-191
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    • 2013
  • Purpose: This study was done to understand the visual system of bat by analyzing the distribution of middle/long (ML) opsin cone photoreceptors in the retina of the greater horseshoe bat. Methods: Experiments have been performed by standard immunocytochemical techniques on retina of the greater horseshoe bat Rhinolophus ferrumequinum. Results: The estimated numbers of ML cones were $27,336{\pm}2,130$ cells and the mean density of them was $7,854{\pm}268cells/mm^2$ among the four retinas. S opsin was appeared a little immunoreactivity in the outer segments of outer nuclear layer of cones. Conclusions: From the well organized spatial distributions of ML opsin and the immunoreactivity of S opsin in the retinas, the greater horseshoe bats have the functions not only reacting in the photopic vision but being able to distinguish the colors.

Periodontal regeneration with nano-hyroxyapatite-coated silk scaffolds in dogs

  • Yang, Cheryl;Lee, Jung-Seok;Jung, Ui-Won;Seo, Young-Kwon;Park, Jung-Keug;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.43 no.6
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    • pp.315-322
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    • 2013
  • Purpose: In this study, we investigated the effect of silk scaffolds on one-wall periodontal intrabony defects. We conjugated nano-hydroxyapatite (nHA) onto a silk scaffold and then seeded periodontal ligament cells (PDLCs) or dental pulp cells (DPCs) onto the scaffold. Methods: Five dogs were used in this study. Bilateral 4 mm${\times}$2 mm (depth${\times}$mesiodistal width), one-wall intrabony periodontal defects were surgically created on the distal side of the mandibular second premolar and the mesial side of the mandibular fourth premolar. In each dog, four of the defects were separately and randomly assigned to the following groups: the PDLCcultured scaffold transplantation group (PDLC group), the DPC-cultured scaffold transplantation group (DPC group), the normal saline-soaked scaffold transplantation group, and the control group. The animals were euthanized following an 8-week healing interval for clinical, scanning electron microscopy (SEM), and histologic evaluations. Results: There was no sign of inflammation or other clinical signs of postoperative complications. The examination of cellseeded constructs by SEM provided visual confirmation of the favorable characteristics of nHA-coated silk scaffolds for tissue engineering. The scaffolds exhibited a firm connective porous structure in cross section, and after PDLCs and DPCs were seeded onto the scaffolds and cultured for 3 weeks, the attachment of well-spread cells and the formation of extracellular matrix (ECM) were observed. The histologic analysis revealed that a well-maintained grafted volume was present at all experimental sites for 8 weeks. Small amounts of inflammatory cells were seen within the scaffolds. The PDLC and DPC groups did not have remarkably different histologic appearances. Conclusions: These observations indicate that nHA-coated silk scaffolds can be considered to be potentially useful biomaterials for periodontal regeneration.

Multi-Junction Space Solar Cell Health Checking Method using Electroluminescence Phenomena (전계발광현상을 이용한 우주용 다접합 태양전지의 건전성 평가기법)

  • Park, Je-Hong;Chang, Young-Keun
    • Journal of the Korean Society for Aeronautical & Space Sciences
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    • v.37 no.10
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    • pp.1017-1026
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    • 2009
  • The solar cell system operates by facing the sun-light. Minor cracks, static discharge, and thermal shock that can happen during production/testing phase can lead to degradation in performance during operation, since solar cells are exposed to extreme thermal/mechanical environment in space. In order to detect small cracks and internal damages in the solar cells due to thermal shocks, which are the core units of a solar cell system, expensive equipment, complicated test process, and much time are required. Therefore, a qualitative method for easily and quickly testing the 'health' of solar cell functionality is required. This dissertation describes a theoretical and technical grounds for quickly and easily evaluating the health of solar cells using electroluminescence effect of Gallium-Arsenide solar cells that are most widely used by spacecrafts in recent years. Also described in the dissertation is the technical issues and constraining factors for applying the proposed method to actual space-rated solar cell systems.

Phototransduction and Visual Cycle in the Ascidian Tadpole Larva

  • Kusakabe, Takehiro;Nakashima, Yuki;Kusakabe, Rie;Horie, Takeo;Kawakami, Isao;Yoshida, Reiko;Inada, Kyoko;Nakagawa, Masashi;Tsuda, Motoyuki
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.37-40
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    • 2002
  • Ascidians are lower chordates, and their tadpole-like larvae share a basic body plan with vertebrates. To study photoreceptive systems in ascidians, we have isolated and characterized cDNA clones for three opsins, five G protein ${\alpha}$ subunits (G${\alpha}$), catalytic and regulatory subunits of cGMP phosphodiesterase (PDE), and arrestin from the ascidian Ciona intestinalis tadpole larva. Ci-opsin1 and Ci-opsin2 are vertebrate-type opsins, while Ci-opsin3 is a retinal photoisomerase similar to retinochrome and mammalian RGR. Both Ci-opsin1 and arrestin are specifically localized in the photoreceptor cells of the ocellus, whereas Ci -opsin2 is not expressed in the photoreceptors, but is co-localized in another population of neurons in the brain with PDE (Ci-PDE9 and Ci-PDE$\delta$). Ci-opsin3 is present in the entire region of the brain. Though five different cDNAs encoding Ga have been cloned, no transducin-type G protein has been found yet. Interestingly, one of G${\alpha}$i isoform is conspicuously expressed in the entire region of the brain. The Ci-opsin3 gene expression was observed in a broad area of the brain vesicle as well as in the visceral ganglion. Genes encoding ascidian homologs of CRALBP and ${\beta}$-CD, whose function is required for the mammalian visual cycle, are co-expressed with Ci-opsin3 in the brain vesicle and visceral ganglion. Localization of Ci-opsin3, CRALBP, and ${\beta}$-CD in a broad area of the brain suggests that the brain of the ascidian larva has a visual cycle system similar to that of the vertebrate RPE. Based on these data, we discuss the evolution of vertebrate visual systems.

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Localization of the Major Retinal Neurotransmitters and Receptors and Müller Glia in the Retina of the Greater Horseshoe Bat (Rhinolophus ferrumequinum) (한국관박쥐 망막의 신경전달물질 및 수용체, 뮬러세포 동정)

  • Lee, Jun-Seok;Kwon, Oh-Ju;Jeon, Tae-Heon;Jeon, Chang-Jin
    • Journal of Korean Ophthalmic Optics Society
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    • v.20 no.3
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    • pp.391-396
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    • 2015
  • Purpose: The objective of this study was to investigate the visual system of the greater horseshoe bat (Rhinolophus ferrumequinum) by location analysis of some major neurotransmitters glutamate, ${\gamma}$-aminobutyric acid (GABA), acetylcholine, and their receptors, and $m{\ddot{u}}ller$ glial cells in retina. Methods: Standard immunocytochemical techniques were used after vibratome section of retinal tissues of adult greater horseshoe bat for this study. Immnoreactions in immunofluorescence images were analyzed using confocal microscope. Results: Anti-glutamate-immunoreactive neurons were mainly localized in the ganglion cell layer (GCL). The majority of anti-GABA-immunoreactive cells distributed in the inner nuclear layer (INL), and GABAA receptors were localized in the inner plexiform layer (IPL). Anti-choline acetyltransferase-immuoreactive cholinergic neurons were mainly located in the INL and GCL, and most of nicotinic acetylcholine receptors were localized in the IPL. The $m{\ddot{u}}ller$ cells in the retina of the greater horseshoe bat stretched theirs range from the GCL to outer nuclear layer (ONL). Conclusions: This study revealed that the retinas of the greater horseshoe bats contain the same major neurotransmitters and receptors, and glial cell in visually functional mammalian retinas. The present results may suggest that the greater horseshoe bats have the functional retinas for visual analysis through the organized retinal neural circuits.

Adhesion and Proliferation Behavior of Retinal Pigment Epithelial Cells on Hesperidin/PLGA Films (헤스페리딘/PLGA 필름에서 망막색소상피세포의 부착과 증식거동)

  • Lee, So Jin;Kang, Su Ji;Kim, Hye Yun;Lee, Jung Hwan;Kim, Eun Young;Kwon, Soon Yong;Chung, Jin Wha;Joo, Choun-Ki;Khang, Gilson
    • Polymer(Korea)
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    • v.38 no.1
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    • pp.24-30
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    • 2014
  • Retinal pigment epithelium (RPE) plays an important role in maintaining the visual function and the degeneration of the RPE causes several retinal degeneration disease. In order to fabricate the suitable carrier for RPE transplantation, the hybrid poly(lactide-co-glycolide) (PLGA) film with hesperidin was prepared. Hesperidin has an anti-inflammatory and antioxidant characteristics. ARPE-19 was seeded on hesperidin/PLGA film and then, cell proliferation was determined by the MTT assay, and cell adhesion and cell morphology were confirmed by SEM. Also, RT-PCR was performed to confirm the expression of the specific genes, and AEC immunohistochemical staining was performed to determine the expression of RPE65. As a result, we confirmed that attachment, proliferation and phenotype maintenance of RPE cells were more excellent on hesperidin/PLGA film than PLGA film, thereby we were able to confirm the potential applications of hesperidin/PLGA film as tissue engineering carrier for regeneration of retina.

In vitro cell recovery methoc as an altermative to human damaged skin recovery test

  • An, Su-Sun;Nam, Ki-Taek;Park, Jong-Ho;Koh, Jae-Sook
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.23 no.3
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    • pp.97-100
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    • 1997
  • These days, the raw materials that have the cell recovering effect are used commonly in cosmetics. In this study, six materials were rested for the characteristics of recovering effect both on vivo and in vitro. Tested raw materials were Soypol, 3-APPA, Apple extract, Polygonatum japonicum extract, Scutellarkd baicalensis extract, Aloe extract. Among these materials, Soypol and 3-APPA were synthesized and others were made by extraction at the Pacific R&D Center. Human forearm skin and cultured skin cell were damaged by sodium lauryl sulfare and then raw materials were applied for open treatment on SLS damaged human skin or cells. The recovering effects of raw materials in vivo were evaluated by measuring transepidermal water loss, skin hydration and erythema and in vitro effects of proliferationg cells were assessed by neutral red uptake assay. In the in vivo study, only the evaluation by TEWL showed correlation with the visual score. Our of six materials, 3-APPA had the most positive effect in both in vivo and in vitro studies and the correlation was r=0.8286 (p=0.042).

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Flexible Visuo-haptic Display (필름형 유연 시-촉각 융합 디스플레이)

  • Yun, Sungryul;Park, Suntak;Park, Bongje;Park, Seung Koo;Nam, Saekwang;Kyung, Ki-Uk;Prahlad, Harsha;von Guggenberg, Philip
    • The Journal of Korea Robotics Society
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    • v.8 no.3
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    • pp.156-163
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    • 2013
  • This paper describes a flexible visuo-haptic display module. We have developed a flexible electro-active polymer (EAP) actuator and a thin flexible visual display with $3{\times}3$ array configuration via polymer technology. The flexible actuator consists of nine EAP cells vertically moving in response to change in their thickness. The flexible display uses polymer based optical waveguide allowing light to scatter only at specific area. The display film is transparent and identically designed to the array pattern to fit for the arrangement of actuator cells. A pressure sensor is installed under the integrated module. The performance of the actuator is proved to be sufficient for satisfying perceivable range of human touch sense. The integrated system can provide interactive haptic feedback such as key pressing, contact vibration sensations, and etc. in accordance with user input.