• Title/Summary/Keyword: cobalt staining

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Expression of Kainate Glutamate Receptors in Type II Cells in Taste Buds of Rats

  • Lee, Sang-Bok;Lee, Cil-Han;Cho, Young-Kyung;Chung, Ki-Myung;Kim, Kyung-Nyun
    • International Journal of Oral Biology
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    • v.33 no.3
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    • pp.83-89
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    • 2008
  • Glutamate-induced cobalt uptake reveals non-NMDA glutamate receptors (GluRs) in rat taste bud cells. Previous studies suggest that glutamate-induced cobalt uptake in taste cells occurs mainly via kainate type GluRs. Cobaltstained cells were immunoreactive against GluR6 and KA1 subunits of GluRs. However, the functions of those type of receptors are not known yet. It is important question which types of taste cells are cobalt-stained when stimulated by glutamate and whether they express these kinds of GluRs. Circumvallate and foliate papilla of Sprague-Dawley rats (45-60 days old) were used. A cobalt-staining technique combined with immunohistochemistry against specific markers for taste bud cell types, such as blood group H antigen (BGH), $\alpha$-gustducin (Gus), or neural cell adhesion molecule (NCAM) was employed. We also performed double labeling of GluR6 or KA1 subunits of GluR with each specific marker for taste bud cell types. Lots of cobaltstained taste bud cells expressed Gus-like immunoreactivity, and subsets of the cobalt stained cells appeared NCAM- or BGH-like immunoreactivity. Stimulation with 1 mM glutamate significantly increased the number of cobaltstained cells in Gus-like immunoreactive cells, but not in NCAM- or BGH-like immunoreactive cells. In the double labeling experiments, GluR6 and KA1 subunits of GluRs were mainly expressed with Gus. These results suggest that kainate glutamate receptors preferentially expressed in type II taste bud cells in rat.

Role of FAK Phosphorylation in Cobalt Chloride-Induced Epithelial-to-Mesenchymal-Like Transition (Cobalt chloride에 의해 유도되는 상피-중간엽 이행에서의 국소부착 단백질의 인산화의 역할 규명)

  • Nam, Ju-Ock
    • Journal of Life Science
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    • v.21 no.2
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    • pp.286-291
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    • 2011
  • Hypoxia is a common condition found in a wide range of solid tumors and is often associated with metastasis and poor clinical outcomes. In the present study, we found that HIF-$1{\alpha}$ was induced by cobalt chloride (500 ${\mu}M$) treatment on human lung cancer cells, A549 and H460, for 24 hr. However, cobalt chloride (500 ${\mu}M$) did not affect cell proliferation of A549 and H460 in 48 hr. Cobalt chloride (500 ${\mu}M$) additionally induced epithelial-to-mesenchymal-like transition (EMT) such as reduced E-cadherin expression and increased ${\alpha}$-SMA expression. These results were confirmed by immunofluorecence experiment in H460 cells. E-cadherin was localized on the outer cell membrane. However, when the cells were treated with 500 ${\mu}M$ cobalt chloride for 24 hr, diffuse E-cadherin staining was observed, characteristic of a migratory mesenchymal phenotype. We also found that cobalt chloride induced integrin ${\beta}3$ expression and FAK phosphorylation in human lung cancer cells using western blotting and FACS anlaysis. Our data suggest that integrin ${\beta}3$-induced FAK phosphorylation may be developed into target molecules for blocking tumor metastasis.

Type II and III Taste Bud Cells Preferentially Expressed Kainate Glutamate Receptors in Rats

  • Lee, Sang-Bok;Lee, Cil-Han;Kim, Se-Nyun;Chung, Ki-Myung;Cho, Young-Kyung;Kim, Kyung-Nyun
    • The Korean Journal of Physiology and Pharmacology
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    • v.13 no.6
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    • pp.455-460
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    • 2009
  • Glutamate-induced cobalt uptake reveals that non-NMDA glutamate receptors (GluRs) are present in rat taste bud cells. Previous studies involving glutamate induced cobalt staining suggest this uptake mainly occurs via kainate type GluRs. It is not known which of the 4 types of taste bud cells express subunits of kainate GluR. Circumvallate and foliate papillae of Sprague-Dawley rats (45~60 days old) were used to search for the mRNAs of subunits of non-NMDA GluRs using RT-PCR with specific primers for GluR1-7, KA1 and KA2. We also performed RT-PCR for GluR5, KA1, $PLC\beta2$, and NCAM/SNAP 25 in isolated single cells from taste buds. Taste epithelium, including circumvallate or foliate papilla, express mRNAs of GluR5 and KA1. However, non-taste tongue epithelium expresses no subunits of non-NMDA GluRs. Isolated single cell RT-PCR reveals that the mRNAs of GluR5 and KA1 are preferentially expressed in Type II and Type III cells over Type I cells.

Effects of $CoCl_2$ on Osteogenic Differentiation of Human Mesenchymal Stem Cells

  • Moon, Yeon-Hee;Son, Jung-Wan;Moon, Jung-Sun;Kang, Jee-Hae;Kim, Sun-Hun;Kim, Min-Seok
    • International Journal of Oral Biology
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    • v.38 no.3
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    • pp.111-119
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    • 2013
  • Objective. To investigate the effects of the hypoxia inducible factor-1 (HIF-1) activation-mimicking agent cobalt chloride ($CoCl_2$) on the osteogenic differentiation of human mesenchymal stem cells (hMSCs) and elucidate the underlying molecular mechanisms. Study design. The dose and exposure periods for $CoCl_2$ in hMSCs were optimized by cell viability assays. After confirmation of $CoCl_2$-induced HIF-$1{\alpha}$ and vascular endothelial growth factor expression in these cells by RT-PCR, the effects of temporary preconditioning with $CoCl_2$ on hMSC osteogenic differentiation were evaluated by RT-PCR analysis of osteogenic gene expression, an alkaline phosphatase (ALP) activity assay and by alizarin red S staining. Results. Variable $CoCl_2$ dosages (up to $500{\mu}M$) and exposure times (up to 7 days) on hMSC had little effect on hMSC survival. After $CoCl_2$ treatment of hMSCs at $100{\mu}M$ for 24 or 48 hours, followed by culture in osteogenic differentiating media, several osteogenic markers such as Runx-2, osteocalcin and osteopontin, bone sialoprotein mRNA expression level were found to be up-regulated. Moreover, ALP activity was increased in these treated cells in which an accelerated osteogenic capacity was also verified by alizarin red S staining. Conclusions. The osteogenic differentiation potential of hMSCs could be preserved and even enhanced by $CoCl_2$ treatment.

Anti-apoptotic effect of fermented Citrus sunki peel extract on chemical hypoxia-induced neuronal injury (화학적 저산소증이 유도하는 뇌신경세포 손상에 있어서 미성숙 진귤 과피 발효 추출물의 보호 효과)

  • Ko, Woon Chul;Lee, Sun Ryung
    • Journal of Nutrition and Health
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    • v.48 no.5
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    • pp.451-456
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    • 2015
  • Purpose: Neuronal apoptotic events induced by aging and hypoxic/ischemic conditions is an important risk factor in neurodegenerative diseases such as ischemia stroke and Alzheimer's disease. The peel of Citrus sunki Hort. ex Tanaka has long been used as a traditional medicine, based on multiple biological activities including anti-oxidant, anti-inflammation, and anti-obesity. In the current study, we examined the actions of fermented C. sunki peel extract against cobalt chloride ($CoCl_2$)-mediated hypoxic death in human neuroblastoma SH-SY5Y cells. Methods: Cell viability was measured by trypan blue exclusion. Expression of apoptosis related proteins and release of cytochrome c were detected by western blot. Production of intracellular reactive oxygen species (ROS) and apoptotic morphology were examined using 2',7'-dichlorofluorescin diacetate (DCF-DA) and 4',6-diamidino-2-phenylindole (DAPI) staining. Results: Exposure to $CoCl_2$, a well-known mimetic agent of hypoxic/ischemic condition, resulted in neuronal cell death via caspase-3 dependent pathway. Extract of fermented C. sunki peel significantly rescued the $CoCl_2$-induced neuronal toxicity with the cell viability and appearance of apoptotic morphology. Cytoprotection with fermented C. sunki peel extract was associated with a decrease in activities of caspase-3 and cleavage of poly (ADP ribose) polymerase (PARP). In addition, increase in the intracellular ROS and release of cytochrome c from mitochondria to the cytosol were inhibited by treatment with extract of fermented C. sunki peel. Conclusion: Based on these data, fermented C. sunki peel extract might have a protective effect against $CoCl_2$-induced neuronal injury partly through generation of ROS and effectors involved in mitochondrial mediated apoptosis.

A study on characteristics of palace wallpaper in the Joseon Dynasty - Focusing on Gyeongbokgung Palace, Changdeokgung Palace and Chilgung Palace - (조선시대 궁궐 도배지 특성 연구 - 경복궁, 창덕궁, 칠궁을 중심으로 -)

  • KIM Jiwon;KIM Jisun;KIM, Myoungnam;JEONG Seonhwa
    • Korean Journal of Heritage: History & Science
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    • v.56 no.1
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    • pp.80-97
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    • 2023
  • By taking wallpaper specimens from Gyeongbokgung Palace, Changdeokgung Palace, and Chilgung Palace preserved from the late Joseon Dynasty to the present, we planned in this study to determine the types and characteristics of the paper used as wallpaper in the Joseon royal family. First, we confirmed the features of paper hanging in the palaces with old literature on the wallpaper used by the royal family based on archival research. Second, we conducted a field survey targeting the royal palaces whose construction period was relatively clear, and analyzed the first layer of wallpaper directly attached to the wall structure after sampling the specimens. Therefore, we confirmed that the main raw material was hanji, which was used as a wallpaper by the royal family, and grasped the types of substances(dyes and pigments) used to produce a blue color in spaces that must have formality by analyzing the blue-colored paper. Based on the results confirmed through the analysis, we checked documents and the existing wallpaper by comparing the old literature related to wallpaper records of the Joseon Dynasty palaces. We also built a database for the restoration of cultural properties when conserving the wallpaper in the royal palaces. We examined the changes in wallpaper types by century and the content according to the place of use by extracting wallpaper-related contents recorded in 36 cases of Uigwe from the 17th to 20th centuries. As a result, it was found that the names used for document paper and wallpaper were not different, thus document paper and wallpaper were used without distinction during the Joseon Dynasty. And though there are differences in the types of wallpaper depending on the period, it was confirmed that the foundation of wallpaper continued until the late Joseon Dynasty, with Baekji(white hanji), Hubaekji(thick white paper), jeojuji(common hanji used to write documents), chojuji(hanji used as a draft for writing documents) and Gakjang(a wide and thick hanji used as a pad). As a result of fiber identification by the morphological characteristics of fibers and the normal color reaction(KS M ISO 9184-4: Graph "C" staining test) for the first layer of paper directly attached to the palace wall, the main materials of hanji used by the royal family were confirmed and the raw materials used to make hanii in buildings of palaces based on the construction period were determined. Also, as a result of analyzing the coloring materials of the blue decorative paper with an optical microscope, ultraviolet-visible spectroscopic analysis(UV-Vis), and X-ray diffraction analysis(XRD), we determined that the type of blue decorative paper dyes and pigments used in the palaces must have formality and identified that the raw materials used to produce the blue color were natural indigo, lazurite and cobalt blue.