• Title/Summary/Keyword: 형광 현미경

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Characteristics and Survival of Genus Vibrio Isolated in the Intertidal Zone of the Yellow Sea near Kunsan (군산인근해역에서 분리동정된 Vibrio 속의 특성과 해수에서의 생존)

  • 왕혜영;이건형
    • Korean Journal of Environmental Biology
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    • v.17 no.4
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    • pp.439-448
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    • 1999
  • To investigate the population dynamics and survival of Genus Vibrio, population densities of aerobic saprophytic bacteria and Vibrio groups were measured 4 times in the intertidal waters of the Yellow Sea near Kunsan from November, 1997 to June, 1998. The distribution of heterotrophic bacteria during the survey periods by plate count and direct count method ranged from 1.2$\pm$0.6$\times$10$^3$~2.0$\pm$1.5$\times$10$^4$CFU ml­$^1$and from 6.0$\pm$4.0$\times$10$^{5}$ ~1.9$\pm$1.5$\times$10$^{7}$ cells ml­$^1$, respectively. Vibrio groups were distributed in the range of 1$\times$10 and 6$\pm$2.2$\times$10$^2$CFU ml­$^1$. The proportion of Vibrio groups to total heterotrophic bacteria was between 0.1 and 6% during the survey periods. A total of 51 isolates was obtained from TCBS agar plates and identified to species level by Biolog Identification System$^{TM}$. As a result, dominant genera were V, mediterranei, V aitguillarum, tr metschnikovii, and V. parahaemolyticus, and isolates were clustered into 26 groups based on the relatedness of average linkage clustering method at 70% level. As for the susceptibility of 51 isolates to 7 kinds of antibacterial agents (gentamicin, ampicillin, chlorarnphenicol, streptomycin, kanamycin, tetracycline, carbenicillin), 96% of isolates showed high resistance to more than one antibiotics and 65% of isolates contained a plasmid, of which size was observed greater than 12 kb, The number of cells of 3 tested strains (V. anguillarum, V. vulnificus, and V. metschnikovii) in filtered aged seawater decreased by approximately 1 to 5 orders of magnitude during 30-d incubation. In most cases, the numbers of cells decreased rapidly until day 3, then decreased slowly by day 30. The number of cells incubated at 15$^{\circ}C$ showed higher survival than those at 4$^{\circ}C$ and $25^{\circ}C$. These results may be considered for the basic supporting data in the risk assessment of vibriosis in summer.r.

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Studies on the Radiation Induced Apoptosis by Morphological and Biochemical Analysis in A431 Cells (방사선에 조사된 인체세포주(A431)에서 형태학적 분석법과 생화학적 분석법을 이용한 방사선 유도 세포고사에 관한 연구)

  • Choi, Keun-Hee;Bom, Hee-Seung;Kim, Ji-Yeul
    • The Korean Journal of Nuclear Medicine
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    • v.33 no.3
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    • pp.306-315
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    • 1999
  • Purpose: We performed this study to evaluate the process of radiation induced apoptosis in A431 skin epithelial cancer cell line. Materials and Methods: Low to high dose radiation (0, 2, 5, 10, 25 Gy) was given to A431 cells by Cs-137 cell irradiator. Apoptosis was evaluated by cell morphology, dye exclusion test, and DNA laddering. Results: Cell viability decreased as the radiation dose increased. Number of apoptotic bodies increased as radiation dose increased. It increased most significantly at 12 hours after irradiation. Lactate dehydrogenase activity in culture medium increased according to radiation dose and time after irradiation. DNA ladders could be identified in irradiated cells, but, it had no correlation with radiation dose or time after irradiation. Conclusion: Radiation-induced apoptosis which was the main course of cell death in A431 cells could be analyzed quantitatively by counting apoptotic bodies under microscope. Apoptosis increased as radiation dose increased.

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Study on the screening method for determination of heavy metals in cellular phone for the restrictions on the use of certain hazardous substances (RoHS) (유해물질 규제법(RoHS)에 따른 휴대폰 내의 중금속 함유량 측정을 위한 스크리닝법 연구)

  • Kim, Y.H.;Lee, J.S.;Lim, H.B.
    • Analytical Science and Technology
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    • v.23 no.1
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    • pp.1-14
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    • 2010
  • It is of importance that all countries in worldwide, including EU and China, have adopted the Restrictions on the use of certain Hazardous Substances (RoHS) for all electronics. IEC62321 document, which was published by the International Electronics Committee (IEC) can have conflicts with the standards in the market. On the contrary Publicly Accessible Specification (PAS) for sampling published by IEC TC111 can be adopted for complementary application. In this work, we tried to find a route to disassemble and disjoint cellular phone sample, based on PAS and compare the screening methods available in the market. For this work, the cellular phone produced in 2001, before the regulation was born, was chosen for better detection. Although X-ray Fluorescence (XRF) showed excellent performance for screening, fast and easy handling, it can give information on the surface, not the bulk, and have some limitations due to significant matrix interference and lack of variety of standards for quantification. It means that screening with XRF sometimes requires supplementary tool. There are several techniques available in the market of analytical instruments. Laser ablation (LA) ICP-MS, energy dispersive (ED) XRF and scanning electron microscope (SEM)-energy dispersive X-ray (EDX) were demonstrated for screening a cellular phone. For quantitative determination, graphite furnace atomic absorption spectrometry (GF-AAS) was employed. Experimental results for Pb in a battery showed large difference in analytical results in between XRF and GF-AAS, i.e., 0.92% and 5.67%, respectively. In addition, the standard deviation of XRF was extremely large in the range of 23-168%, compared with that in the range of 1.9-92.3% for LA-ICP-MS. In conclusion, GF-AAS was required for quantitative analysis although EDX was used for screening. In this work, it was proved that LA-ICP-MS can be used as a screening method for fast analysis to determine hazardous elements in electrical products.

induction of Apoptosis in Human Cancer Cells with Compositae Extracts (국화과 추출물의 암세포 증식 억제 효과)

  • An, In-Jung;Kwon, Jung-Ki;Lee, Jin-Seok;Park, Ha-Seung;Kim, Dong-Chan;Choi, Byung-Jun;Lee, Kyu-Min;Park, Youg-Jin;Jung, Ji-Youn
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.5
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    • pp.584-590
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    • 2012
  • Dried $Compositae$ flowers have traditionally been used for the treatment of anti-inflammatory and anti-oxidative stress in Korea. This paper investigates the effects of $Compositae$ extracts on the inhibition of proliferation and apoptosis of human gastric cancer AGS cells, human breast cancer MDA-MB-231 cells, and SK-BR-3 cells. The proliferation of AGS cells, MDA-MB-231 cells, and SK-BR-3 cells were determined by MTT assay. Several $Compositae$ extracts inhibited proliferation of AGS cells, MDA-MB-231 cells, and SK-BR-3 cells in a dose-dependent manner. To assess the apoptosis of $Compositae$ extracts, the nuclei of MDA-MB-231 cells were stained with DAPI. The presence of chromatin condensation in the $Compositae$ extract-treated cells was detected on a fluorescent microscope (${\times}200$). We conducted Western blot analysis of changes in Bcl-2, Bax, and p53 protein expression levels. Apoptosis by $Chrysanthemum$ $zawadskii$ subsp. coreanum, $Chrysanthemum$ $zawadskii$ var. $tenuisectum$ and $Rudbeckia$ $laciniata$ var. $hortensis$ treatment created a decrease in Bcl-2 expression, whereas the expression of Bax and p53 were increased. These results indicate that $Chrysanthemum$ $zawadskii$ $subsp.$ $coreanum$, $Chrysanthemum$ $zawadskii$ var. $tenuisectum$ and $Rudbeckia$ $laciniata$ var. $hortensis$ inhibit breast cancer cell growth through the induction of apoptosis.

Technique to Generate Sterile Males of Striped Fruit Flies, Zeugodacus scutellata, using Electron Beam Irradiation and their Application to Genetic Control (전자빔 조사를 이용한 호박꽃과실파리의 불임 수컷 제조 기술과 유전방제 응용)

  • Kim, Yonggyun;Al Baki, Md. Abdullah;Kwon, Gimyeon
    • Korean journal of applied entomology
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    • v.59 no.1
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    • pp.25-35
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    • 2020
  • The striped fruit fly, Zeugodacus scutellata, is endemic in Korea, but it has been regarded as one of the serious quarantine pests throughout the world. Sterile insect release technique (SIT) has been used to eradicate quarantine fruit flies. This study developed a technique to generate sterile males and applied SIT to control Z. scutellata. First of all, the reproductive systems of Z. scutellata adults were examined with fluorescent microscope. Polytrophic ovaries comprises of around 100 follicles with developing oocytes. Each follicle contains an oocyte with several nurse cells and are surrounded with follicular epithelium. Oocyte development began at 10 days after adult emergence (DAE) and formed chorionated oocytes after 20 DAE. On the other hand, male testes were well developed just after adult emergence. The vas deferens was filled with motile sperms. To generate sterile males, different doses (0~1,000 Gy) doses of electron beam were irradiated to 3~5 days old pupae of Z. scutellata. When male pupae were irradiated with electron beam at 200 Gy, they developed and mated with females without any significant difference compared to untreated males. Although the untreated females mated with the 200 Gy-irradiated males laid eggs, no eggs did not hatch. The 200 Gy-irradiated males were then applied to untreated male and female flies in a density ratio of 1:9 (untreated males : treated males). The laid eggs suffered significant infertility. These results suggest that electron beam-irradiated pupae at 200 Gy resulted in male sterility and the resulting males would be applied to SIT.

Studies on the Anticancer Effect of Apigenin in KB Cell Xenograft Nude Mouse Model (구강암 세포주를 이종 이식한 누드마우스에서 apigenin의 경구투여에 따른 항암효능에 관한 연구)

  • Lee, Jin-Seok;Seo, Hyeong-Seok;Kim, So-Jung;Kim, Hyeong-Jin;Kim, Jin;Lee, Seung-Ho;Park, Young-Seok;Park, Byung-Kwon;Kim, Byeong-Soo;Kim, Sang-Ki;Jung, Ji-Youn
    • Journal of Life Science
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    • v.20 no.10
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    • pp.1519-1524
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    • 2010
  • Apigenin (4', 5, 7-trihydroxyflavone), a common dietary flavonoid abundantly present in fruits and vegetables, has shown remarkable anti-proliferative effects against various malignant cell lines. To observe the anti-proliferative effects, oral cavity cancer cell lines, $6{\times}10^3$ cells/well (96 well plate) of KB oral cavity tumor cells were plated and 24 hr later treated with apigenin for one day, after which MTT assay was performed. Apigenin induced cell death in a dose-dependent manner after incubation. Cell viability was significantly decreased in the group treated with 100 ${\mu}M$ apigenin for 24 hr (p<0.05) compared to the control group. To assess apoptosis, the nuclei of KB cells were stained with DAPI. The presence of chromatin condensation in the apigenin treated cells was detected on a fluorescent microscope (${\times}200$). We investigated the in vivo growth inhibitory effects of apigenin on oral cavity cancer KB tumor xenograft subcutaneously implanted in male nude mice. Apigenin was administered to mice by gavage at doses of 25 and 50 mg/kg/day in 0.2ml of PBS. Tumor volume was significantly decreased in 25 and 50 mg/kg apigenin-administration groups compared to the control group. For apoptosis analysis, TUNEL staining was performed. A significant increase in TUNEL positive cells was found in the 25 mg/kg apigenin administration group compared to the non- apigenin administration group. Histopathological changes were not observed. These results indicate that apigenin inhibits oral cavity cancer cell growth through the induction of apoptosis.

Coexpression of $P2X_3$ with TRPV1 in the Rat Trigeminal Sensory Nuclei (흰쥐 삼차신경감각핵에서 $P2X_3$와 TRPV1의 공존에 관한 연구)

  • Moon, Yong-Suk;Ryoo, Chang-Hyun;Cho, Yi-Sul;Kim, Hong-Tae;Park, Mae-Ja;Paik, Sang-Kyoo;Moon, Che-Il;Kim, Yun-Sook;Bae, Yong-Chul
    • Applied Microscopy
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    • v.38 no.3
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    • pp.151-157
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    • 2008
  • Trigeminal primary afferents expressing $P2X_3$ or transient receptor potential vanilloid 1 (TRPV1) are involved in the transmission of nociceptive information. In order to characterize $P2X_3$- and TRPV1-immunopositive neurons in the trigeminal ganglion (TG) and trigeminal caudal nucleus (Vc), we performed immunofluorescence experiments using anti-$P2X_3$ and anti-TRPV1 antisera and a morphometric analysis. 77.4% (1,401/1.801) of all the $P2X_3$-postive neurons coexpressed TRPV1 and 51.9% (1,401/2,698) of all the THFV1-immunopositive neurons also costained for $P2X_3$ in the TG. Immunoreactivity for both $P2X_3$ and TRPV1 were present in medium-sized neurons but not in small- and large-sized neurons. $P2X_3$ and/or TRPV1-immunopositive fibers were observed in the primary afferents and their associated axons in the Vc. These fibers and terminals were distributed in the superficial lamina of Vc: $P2X_3$-immunopositive fibers and terminals were distributed in the lamina I and II, expecially in the inner part of lamina II (lamina IIi), whereas TRPV1-immunopositive ones were densely detected in the lamina I and outer part of lamina II (lamina IIo). Immunopositive fibers and terminals for both $P2X_3$ and TRPV1 were observed on the border between lamina IIi and IIo. These results suggest that terminals coexpressing $P2X_3$ and TRPV1 are involved in specific roles in the transmission and processing of orofacial nociceptive information.

Characterization of Nitric Oxide (NO)-Induced Cell Death in Lung Epithelial Cells (폐상피세포에서 Nitric Oxide (NO)에 의한 세포사에 관한 연구)

  • Yong, Wha Shim;Kim, Youn Seup;Park, Jae Seuk;Jee, Young Koo;Lee, Kye Young
    • Tuberculosis and Respiratory Diseases
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    • v.56 no.2
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    • pp.187-197
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    • 2004
  • Background : Nitric Oxide (NO) is a multi-faceted molecule with dichotomous regulatory roles in many areas of biology. NO can promote apoptosis in some cells, whereas it inhibits apoptosis in other cell types. This study was performed to characterize NO-induced cell death in lung epithelial cells and to investigate the roles of cell death regulators including iron, bcl-2 and p53. Methods : A549 cells were used for lung epithelial cells. SNP (sodium nitroprusside) and SNAP (S-nitroso-N-acetyl- penicillamine) were used for NO donor. Cytoxicity assay was done by MTT assay and crystal violet assay. Apoptotic assay was done by fluorescent microscopy after double staining with propidium iodide and hoecst 33342. Iron inhibition study was done with RBCs and FeSO4. For bcl-2 study, bcl-2 overexpressing cells (A549-bcl-2) were used and for p53 study, Western blot analysis and p53 functionally knock-out cells (A549-E6) were used. Results : SNP and SNAP induced dose-dependent cell death in A549 cells and fluorescent microscopy revealed that SNAP induced apoptosis in low doses but necrosis in high doses while SNP induced exclusively necrotic cell death. Iron inhibition study using RBCs and FeSO4 significantly blocked SNAP-induced cell death. And also SNAP-induced cell death was blocked by bcl-2 overexpression. Finally, we found that SNAP activate p53 by Western blot analysis and that SNAP-induced cell death was decreased in the abscence of p53. Conclusion : In lung epithelial cells, NO can induce cell death, more precisely apoptosis in low doses and necrosis in high doses. And iron, bcl-2, and p53 play important roles in NO-induced cell death.

A Study on the Presence of Murals by Scientific Investigation on the Inner Walls of West Ancient Tomb No.1 and 2 Neungsan-ri, Buyeo (부여 능산리 서고분군 1·2호분 내벽의 과학적 조사를 통한 벽화 존재 유무 연구)

  • Lee, Hanhyoung;Kim, Dongwon;Lee, Hwasoo
    • Conservation Science in Museum
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    • v.22
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    • pp.41-52
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    • 2019
  • Inner walls of the stone chamber of West Ancient Tomb No. 1 and 2 in Neungsan-ri, Buyeo-gun have been inspected for possible trace of murals. Tomb No.1 has a rough surface finish of the stone wall and no traces of murals was observed in any part of the stone walls except the ceiling part of the main chamber. On the ceiling surface, there is black colored area, which showed same visual characteristics for both the surface and interior upon slight scratch of the surface, suggesting that it may not be a painted layer. In addition, this black material is not artificial stuff like black ink but is confirmed as biotite from X-ray diffraction analysis that is one of the constituents of the stone wall. In case of tomb No. 2, white material, that is confirmed as lime(calcite, CaCO3) by X-ray diffraction analysis, was observed on the wall surface of the east, west and north, suggesting possible existence of murals. The lime layers, however, are located mostly on the entrance of east wall of main chamber and the place of passage whereas they are observed only in lower parts on the other walls. It may have been formed by the inflow of soil and lime from the outside as the form of the lime layer in the east wall corresponds to the traces of soil and lime deposited from the thief pit. Furthermore, the filling material found in the gap between the stone slabs of the four directions and the ceiling was confirmed as clay soil, which is different material from the lime present on the stone wall surface. If the lime layer had been artificially constructed for the purpose of creating murals, it would have been more reasonable to use lime as well in the gap between the stone slabs of the four directions and ceiling. In this regard, we conclude that there are no murals in the Tomb No. 2 in the Neungsan-ri.

Analysis of the Role of RGG box of human hnRNP A1 protein (인간 hnRNP A1 단백질에 포함된 RGG 상자의 기능 분석)

  • Choi, Mieyoung
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.12
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    • pp.575-580
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    • 2017
  • This study analyzed the effects of RGG box of hnRNP A1 on its subcellular localization and stabilization of hnRNP A1 over a three year period from October 2014. First, a 6R/K mutation in RGG box was generated, and pcDNA1-HA-hnRNP A1(6R/K) was constructed. The subcellular localization of hnRNP A1(6R/K) from the HeLa cells transfected with this plasmid DNA was analyzed by immunofluorescence microscopy. HA-hnRNP A1(6R/K) was found to exhibit nuclear and cytoplasmic fluorescence. The stability of hnRNP A1(6R/K) was checked by Western blot analysis using the expressed protein from the HeLa cells transfected with the pcDNA1-HA-hnRNP A1(6R/K). The results show that HA-hnRNP A1(6R/K) has a smaller size. These confirm that HA-hnRNP A1(6R/K) is localized both in the nuclear and cytoplasm, not because 6R/K mutation affects the nuclear localization of hnRNP A1, but because 6R/K mutation causes hnRNP A1(6R/K) to cleave at the mutation or near the mutation site. The cleaved protein fragment, which lacks the M9 domain (i.e. nuclear localization signal of hnRNP A1), did not exhibit nuclear fluorescence. This suggests that the arginines of RGG box in hnRNP A1 play an important role in stabilizing hnRNP A1. An analysis of the RNA-binding ability of hnRNP A1(6R/K) expressed and purified from bacteria will be a subsequent research project.