• Title/Summary/Keyword: Confocal Image

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Monitoring microRNAs Using a Molecular Beacon in CD133+/CD338+ Human Lung Adenocarcinoma-initiating A549 Cells

  • Yao, Quan;Sun, Jian-Guo;Ma, Hu;Zhang, An-Mei;Lin, Sheng;Zhu, Cong-Hui;Zhang, Tao;Chen, Zheng-Tang
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.161-166
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    • 2014
  • Lung cancer is the most common causes of cancer-related deaths worldwide, and a lack of effective methods for early diagnosis has greatly impacted the prognosis and survival rates of the affected patients. Tumor-initiating cells (TICs) are considered to be largely responsible for tumor genesis, resistance to tumor therapy, metastasis, and recurrence. In addition to representing a good potential treatment target, TICs can provide clues for the early diagnosis of cancer. MicroRNA (miRNA) alterations are known to be involved in the initiation and progression of human cancer, and the detection of related miRNAs in TICs is an important strategy for lung cancer early diagnosis. As Hsa-miR-155 (miR-155) can be used as a diagnostic marker for non-small cell lung cancer (NSCLC), a smart molecular beacon of miR-155 was designed to image the expression of miR-155 in NSCLC cases. TICs expressing CD133 and CD338 were obtained from A549 cells by applying an immune magnetic bead isolation system, and miR-155 was detected using laser-scanning confocal microscopy. We found that intracellular miR-155 could be successfully detected using smart miR-155 molecular beacons. Expression was higher in TICs than in A549 cells, indicating that miR-155 may play an important role in regulating bio-behavior of TICs. As a non-invasive approach, molecular beacons could be implemented with molecular imaging to diagnose lung cancer at early stages.

Analysis of metabolites in wheat roots in response to salinity stress

  • Kim, Da-Eun;Roy, Swapan Kumar;Kim, Ki-Hyun;Cho, Seong-Woo;Park, Chul-Soo;Lee, Moon-Soon;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.200-200
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    • 2017
  • Salinity stress is one of the most important abiotic stresses and severely impairs plant growth and production. Root is the first site for nutrient accumulation like as $Na^+$ in the plant. To investigate the response of wheat root under salinity stress, we executed the characterization of morphology and analysis of metabolites. Wheat seeds cv. Keumgang (Korean cultivar) were grown on the moist filter paper in Petri dish. After 5 days, seedlings were transferred to hydroponic apparatus at 1500 LUX light intensity, at $20^{\circ}C$ with 70% relative humidity in a growth chamber. Seedlings (5-day-old) were exposed to 50mM, 75mM, 100mM NaCl for 5 days. Ten-day-old seedlings were used for morphological characterization and metabolite analysis. Root and leaf length became shorter in high NaCl concentration compared to following NaCl treatment. For confirmation of salt accumulation, wheat roots were stained with $CoroNa^+$ Green AM, and fluoresce, and the image was taken by confocal microscopy. $Na^+$ ion accumulation rate was higher at 100mM compared to the untreated sample. Furthermore, to analyze metabolites in the wheat root, samples were extracted by $D_2O$ solvent, and extracted sample was analyzed by 1H NMR spectroscopy. Fourteen metabolites were identified in wheat roots using NMR spectroscopy. Methanol and ethanol were up-regulated, whereas formate, aspartate, aminobutyrate, acetate and valine were down-regulated under salinity stress on roots of wheat. Fumarate had no change, while glucose, betaine, choline, glutamate and lactate were unevenly affected during salinity stress.

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In Situ Observation of Solidification Behavior in Undercooled $Pd_{40}Cu_{30}Ni_{10}P_{20}$ Alloy Melts during Linear Cooling (연속냉각 중 과냉 된 $Pd_{40}Cu_{30}Ni_{10}P_{20}$ 합금 용탕의 실시간 응고거동 관찰)

  • Kim, Ji-Hun
    • Journal of Korea Foundry Society
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    • v.23 no.5
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    • pp.276-285
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    • 2003
  • In the undercooled melt of $Pd_{40}Cu_{30}Ni_{10}P_{20}$ alloy, the solidification behavior including nucleation and growth of crystals at the micrometer level has been observed in-situ by use of a confocal scanning laser microscope combined with an infrared image furnace. The $Pd_{40}Cu_{30}Ni_{10}P_{20}$ alloy specimens were cooled from the liquid state to glass transition temperature. 575 K, at various cooling late under a helium gas flow. According to the cooling rate, the morphologies of the solidification front are changed among various types, irregular jog like front, columnar dendritic front, cellular grain, star like shape jog and fine grain, etc. The velocities of the solid-liquid interface are measured to be $10^{-5}{\sim}10^{-8}$ m/s which are at least two orders higher than the theoretical crystal growth rates. Combining the morphologies observed in terms of cooling rates and their solidification behaviors, we conclude that phase separation takes place in the undercooled molten $Pd_{40}Cu_{30}Ni_{10}P_{20}$ alloy. The continuous cooling transformation (CCT) diagram was constructed from solidification onset time at various linear cooling conditions with different rate. The CCT diagram suggests that the critical cooling rate for glassy solidification is about 1.5 K/s, which is in agreement with the previous calorimetric findings.

Protein Disulfide Isomerase Is Cleaved by Caspase-3 and -7 during Apoptosis

  • Na, Kyung Sook;Park, Byoung Chul;Jang, Mi;Cho, Sayeon;Lee, Do Hee;Kang, Sunghyun;Lee, Chong-Kil;Bae, Kwang-Hee;Park, Sung Goo
    • Molecules and Cells
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    • v.24 no.2
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    • pp.261-267
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    • 2007
  • Apoptotic signals are typically accompanied by activation of aspartate-specific cysteine proteases called caspases, and caspase-3 and -7 play crucial roles in the execution of apoptosis. Previously, using the proteomic approach, protein disulfide isomerase (PDI) was found to be a candidate substrate of caspase-7. This abundant 55 kDa protein introduces disulfide bonds into proteins (via its oxidase activity) and catalyzes the rearrangement of incorrect disulfide bonds (via its isomerase activity). PDI is abundant in the ER but is also found in non-ER locations. In this study we demonstrated that PDI is cleaved by caspase-3 and -7 in vitro. In addition, in vivo experiment showed that it is cleaved during etoposide-induced apoptosis in HL-60 cells. Subcellular fractionation showed that PDI was also present in the cytosol. Furthermore, only cytosolic PDI was clearly digested by caspase-3 and -7. It was also confirmed by confocal image analysis that PDI and caspase-7 partially co-localize in both resting and apoptotic MCF-7 cells. Overexpression of cytosolic PDI (ER retention sequence deleted) inhibited cell death after an apoptotic stimulus. These data indicate that cytosolic PDI is a substrate of caspase-3 and -7, and that it has an anti-apoptotic action.

COMPARATIVE STUDY ON THE EARLY DETECTION OF ENAMEL LESIONS USING DIFOTI AND LASER FLUORESCENCE (Digital Imaging Fiber-Optic Trans-Illumination과 Laser Fluorescence를 이용한 법랑질 우식증의 조기 진단에 관한 비교 연구)

  • Maeng, Myoung-Ho;Kim, Seung-Oh;Kim, Jong-Soo
    • Journal of the korean academy of Pediatric Dentistry
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    • v.33 no.2
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    • pp.207-220
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    • 2006
  • The newly developed equipments for the early detection of carious lesion are LFD (laser fluorescence device), Ultrasonic diagnostic system, CLSM(confocal laser scanning microscopy), QLF(quantitative light-induced fluorescence) and DIFOTI (digital imaging fiber-optic trans-illumination) system. In this study, DIFOTI system and LFD were used for the detection of early enamel caries. Twenty five primary teeth extracted from twenty one children at around the dentitional exchanging period were selected as samples. The results obtained from DIFOTI imaging and LFD measurement were compared with those of CLSM and comprehensive evaluations were made for the diagnostic capacity of each device. In vitro test, 40 sample teeth with their buccal & lingual surface formed by a window of $2{\times}3mm$ in diameter were immersed in artificial demineralizing solution for the period of 4, 8, 12 and 16 days. The results obtained from the experimental groups (DIFOTI, LFD) were compared to control group (CLSM) and we have reached to the following conclusions. 1. The sensitivity and specificity of DIFOTI system operated in oral environment was 88.2% and 76.9% respectively. 2. The sensitivity and specificity of LFD measured in oral environment was 76.5% and 69.2% respectively. 3, Regression analysis on the light transparent rate of DIFOTI showed its decrease according to the length of primary enamel decalcification performed in vitro(r=-0.96, p<0.05). 4. No statistically significant difference between LFT measurement and the length of in vitro decalcification was found in regression analysis (p>0.05). 5. The correlation coefficient of DIFOTI image transparent rate and the lesion depth of CLMS was -0.6988 (p<0.05), whereas no statistically significant difference was found for LFD measurement.

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Design of an Endoscopic Microscope Objective Lens Composed of Flexible Fiber Bundle and Gradient-index with a High Resolution and a Minimally-Invasive Outer Diameter (광섬유 다발과 Gradient-index Lens가 결합된 고 분해능 및 최소침습 직경의 공초점 내시 현미경 대물렌즈의 설계)

  • Jang, Sun-Young;Rim, Cheon-Seog
    • Korean Journal of Optics and Photonics
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    • v.19 no.2
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    • pp.87-94
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    • 2008
  • We present a new design for an endoscope objective lens composed of a lexible fiber bundle with 30,000 core, and a gradient-index (GRIN) objective lens with an optical adaptor. The characteristic of this objective lens is to be minimally-invasive to be able to insert easily in the internal organs of live animals. The GRIN lens has a small diameter and a very simple construction, which is selected with the diameter of 1.0 mm and numerical aperture of 0.5 to achieve a minimally-invasive outer diameter and a high resolution. The resultant designed lens shows the performance as follows; a lateral resolution of 1.63 um and diameters of 100% encircled energy of $0.3\;{\mu}m$ and $0.83\;{\mu}m$ for the on-axis and the off-axis image point, respectively. Also, we can present a cheap solution with a lateral resolution of 1.74 um and diameters of 100% encircled energy of $1.10\;{\mu}m$ and $2.84\;{\mu}m$ for the on-axis and the off-axis image point, respectively.

Spectroscopic Comparison of Photo-oxidation of Outside and Inside of Hair by UVB Irradiation (자외선B 조사에 의한 모발 외부와 내부의 광산화에 관한 분광학적 비교)

  • Ha, Byung-Jo
    • Applied Chemistry for Engineering
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    • v.31 no.2
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    • pp.220-225
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    • 2020
  • Hair is made of proteins containing various amino acids. Ultraviolet (UV) radiation is believed to be responsible for the most damaging effects of sunlight, and also plays an important role in hair aging. The purpose of this study was to investigate the changes in morphological and chemical structures after ultraviolet B (UVB) irradiation of human hair. The UVB-irradiated hair showed characteristic morphological and structural changes, compared to those of the normal hair. The result from a scanning electron microscope (SEM) equipped with an energy dispersive X-ray diffractometer (EDX) showed that the scale of UV-irradiated hair appeared to be rough and the amount of oxygen element was higher than that of the normal hair. Fluorescence and three dimensional (3D) topographical images were obtained by a confocal laser scanning microscope (CLSM). In 3D images, the green emission intensity of normal hair was much higher than that of fluorescing UVB-irradiated hair. The intensity of green emission reflects the intrinsic fluorescence of hair protein. Also, a fluorescent imaging method using fluorescamine reagent was used to identify the free amino groups resulting from a peptide bond breakage in UVB-irradiated hair. Strong blue fluorescence of UVB-irradiated hair, which indicates a very high level of amino groups, was observed by CLSM. Therefore, the fluorescamine as an extrinsic fluorescence could provide a useful tool to identify the peptide bond breakage in UVB-irradiated hair. Infrared image mapping was also employed to assess the cross-sections of normal and UVB-irradiated specimens to examine the oxidation of disulfide bonds. The degree of peak areas with strong absorbance for the disulfide mono-oxide was spread from the outside to the inside of hair. The spectroscopic techniques used alone, or in combination, launch new possibilities in the field of hair cosmetics.

Acute Cytotoxicity Testing of Polyhexamethylene-biguanide (PHMG) and Epigallocatechin-gallate (EGCG) Mixture on the Cultured Human Corneal Epithelial Cell (보존제 PHMB(polyhexamethylene biguanide)-EGCG(epigallocatechin gallate) 혼합물의 각막상피세포 급성독성 평가)

  • Kim, Nam-youl;Lee, Koon-Ja
    • The Korean Journal of Vision Science
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    • v.20 no.4
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    • pp.531-541
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    • 2018
  • Purpose : To evaluate the acute cytotoxic effect of polyhexamethylene biguanide (PHMB), epigallocatechin gallate (EGCG) and PHMB/EGCG mixture on the cultured human corneal epithelial cell (HCEpiC). Methods : HCEpiCs were cultured in the media of HCEpiC containing 0.00001~0.005% PHMB, 0.001~5% EGCG and 0.00005% PHMB/0.05% EGCG mixture respectively for 30, 60, 120 and 240 min. Cultured HCEpiCs were fixed and stained with Draq5 and cell viability and apoptosis were evaluated using confocal microscope and ImageXpress $Ultra^{TM}$. Results : Cultured HCEpiC did not show cytotoxic effect at below 0.00005% PHMB and below 0.05% EGCG concentration. In the media containing 0.00005% PHMB/0.05% EGCG, acute cytototoxic effect was not found, whereas damaged HCEpiCS were increased and survival cells were decreased in the media incubated for 240 min. Conclusion : The mixture of 0.00005% PHMB/0.05% EGCG showed non acute cytotoxic effect on the cultured HCEpiCs, however it is needed to investigate its chronic cytotoxic effect.

Induction of Apoptosis by Citrus grandis Osbeck Peel (CGP) Extract in HL60 Cells (당유자 과피 추출물에 의한 HL60 세포의 Apoptosis 유도)

  • Hyon, Jae-Seok;Kang, Sung-Myung;Kim, Areum-Daseul;Oh, Myung-Cheol;Oh, Chang-Kyung;Kim, Dong-Woo;Jeon, You-Jin;Kim, Soo-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.10
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    • pp.1317-1323
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    • 2009
  • In the present study, we investigated the anti-proliferation activity of Citrus grandis Osbeck peel (CGP) in HL60 (human promyelocytic leukemia) cells. It was found that 80% ethanol extract of CGP could inhibit the cell growth in a dose-dependent manner ($250{\sim}1,000{\mu}g/mL$), which was associated with morphological changes and apoptotic cell death such as depolarized mitochondrial membrane, formation of apoptotic bodies and increased populations of apoptotic sub-G1 phase. The results indicate that CGP extract inhibits the growth of HL60 cancer cells by the induction of apoptosis, which may be mediated by its ability to change the Bcl family proteins and increase the activation of caspase-3 and PARP. Therefore, it is suggested that CGP has the potential to provide a remarkable natural defense against the proliferation of HL60 cells.

Aluminum toxicity-induced alterations of root proteome in wheat seedlings

  • Oh, Myeong Won;Roy, Swapan Kumar;Cho, Kun;Cho, Seong-Woo;Park, Chul-Soo;Chung, Keun-Yook;Choi, Jong-Soon;Woo, Sun-Hee
    • Proceedings of the Korean Society of Crop Science Conference
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    • 2017.06a
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    • pp.127-127
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    • 2017
  • Aluminum is the most abundant metallic element in the Earth's crust and considered as the most limiting factor for plant productivity in acidic soils. The inhibition of root growth is recognized as the primary effect of Al toxicity. Seeds of wheat cv. Keumkang (Korean cultivar) were germinated on petridish for 5 days and then transferred hydroponic apparatus which was treated with $0{\mu}M$ $AlCl_3$ (control), $100{\mu}M$ $AlCl_3$ and $150{\mu}M$ $AlCl_3$ for 5 days. The length of roots, shoots and fresh weight of wheat seedlings were decreased under aluminum stress. The concentrations of $K^+$, $Mg^{2+}$ and $Ac^{2+}$ were decreased whereas $Al^{3+}$ and $P_2O_5{^-}$ concentration was increased under aluminum stress. Using confocal microscopy, the fluorescence intensity of aluminum was increased with morin staining. In this study, a proteome analysis was performed to identify proteins, which is responsible to aluminum stress in wheat roots. In 10-day-old seedlings, proteins were extracted from roots and separated by 2-DE, stained by CBB. Using image analysis, a total of 47 differentially expressed protein spots were selected, whereas 19 protein spots were significantly up-regulated such as s-adenosylmethionine, oxalate oxidase, malate dehydrogenase, cysteine synthase, ascorbate peroxidase and 28 protein spots were significantly down-regulated such as heat shock protein 70, o-methytransferase 4, enolase, amylogenin by aluminum stress following protein spots analyzed by LTQ-FTICR mass spectrometry. The results provide the global picture of Al toxicity-induced alterations of protein profiles in wheat roots, and identify the Al toxicity-responsive proteins related to various biological processes that may provide some novel clues about plant Al tolerance.

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