• Title/Summary/Keyword: Epithelial Cell

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Examination of the xanthosine response on gene expression of mammary epithelial cells using RNA-seq technology

  • Choudhary, Shanti;Li, Wenli;Bickhart, Derek;Verma, Ramneek;Sethi, R.S.;Mukhopadhyay, C.S.;Choudhary, Ratan K.
    • Journal of Animal Science and Technology
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    • v.60 no.7
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    • pp.18.1-18.12
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    • 2018
  • Background: Xanthosine treatment has been previously reported to increase mammary stem cell population and milk production in cattle and goats. However, the underlying molecular mechanisms associated with the increase in stem cell population and milk production remain unclear. Methods: Primiparous Beetal goats were assigned to the study. Five days post-partum, one mammary gland of each goat was infused with xanthosine (TRT) twice daily ($2{\times}$) for 3 days consecutively, and the other gland served as a control (CON). Milk samples from the TRT and CON glands were collected on the 10th day after the last xanthosine infusion and the total RNA was isolated from milk fat globules (MEGs). Total RNA in MFGs was mainly derived from the milk epithelial cells (MECs) as evidenced by expression of milk synthesis genes. Significant differentially expressed genes (DEGs) were subjected to Gene Ontology (GO) terms using PANTHER and gene networks were generated using STRING db. Results: Preliminary analysis indicated that each individual goat responded to xanthosine treatment differently, with this trend being correlated with specific DEGs within the same animal's mammary gland. Several pathways are impacted by these DEGs, including cell communication, cell proliferation and anti-microbials. Conclusions: This study provides valuable insights into transcriptomic changes in milk producing epithelial cells in response to xanthosine treatment. Further characterization of DEGs identified in this study is likely to delineate the molecular mechanisms of increased milk production and stem or progenitor cell population by the xanthosine treatment.

The Protective Effect of Quercetin-3-O-${\beta}$-D-Glucuronopyranoside on Ethanol-induced Damage in Cultured Feline Esophageal Epithelial Cells

  • Cho, Jung-Hyun;Park, Sun-Young;Lee, Ho-Sung;Whang, Wan-Kyunn;Sohn, Uy-Dong
    • The Korean Journal of Physiology and Pharmacology
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    • v.15 no.6
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    • pp.319-326
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    • 2011
  • Quercetin-3-O-${\beta}$-D-glucuronopyranoside (QGC) is a flavonoid glucoside extracted from Rumex Aquaticus Herba. We aimed to explore its protective effect against ethanol-induced cell damage and the mechanism involved in the effect in feline esophageal epithelial cells (EEC). Cell viability was tested and 2',7'-dichlorofluorescin diacetate assay was used to detect intracellular $H_2O_2$ production. Western blotting analysis was performed to investigate MAPK activation and interleukin 6 (IL-6) expression. Exposure of cells to 10% ethanol time-dependently decreased cell viability. Notably, exposure to ethanol for 30 min decreased cell viability to 43.4%. When cells were incubated with $50{\mu}M$ QGC for 12 h prior to and during ethanol treatment, cell viability was increased to 65%. QGC also inhibited the $H_2O_2$ production and activation of ERK 1/2 induced by ethanol. Pretreatment of cells with the NADPH oxidase inhibitor, diphenylene iodonium, also inhibited the ethanol-induced ERK 1/2 activation. Treatment of cells with ethanol for 30 or 60 min in the absence or presence of QGC exhibited no changes in the IL-6 expression or release compared to control. Taken together, the data indicate that the cytoprotective effect of QGC against ethanol-induced cell damage may involve inhibition of ROS generation and downstream activation of the ERK 1/2 in feline EEC.

Growth and Differentation of Rat Mammary Epithelial Cells Cultured in Serum-free Medium

  • Kim, Dong-Yeum;Jhun, Byung-Hak;Lee, Kyung-Hee;Hong, Seung-Chul;Clifton, Kelly-H.;Kim, Nam-Deuk
    • Archives of Pharmacal Research
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    • v.20 no.4
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    • pp.297-305
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    • 1997
  • A new serum-free defined medium was developed that supports the growth of normal rat mammary epithelial cells. Mammary organoids from the glands of female F344 rats were cultured in a serum-free medium. Monolayer culture colonies developed within a week and remained viable for months in culture. Upon subculture of one-week-old primary colonies, almost the same morphology of colonies was developed. The scrape loading/dye transfer technique showed that most of colonies that developed in a serum-free medium containing EGF, human transferrin, insulin, and hydrocortisone (basal serum-free medium, BSFM) failed to show cell-cell communication. However, colonies cultured in BSFM supplemented with prolactin, $E_2$, and progesterone (complete hormone serum-free medium, CHSFM) showed cell-cell communication at 14 days of primary culture or of subculture. By flow cytometry with FITCPNA and PE-anti-Thy-1.1 monoclonal antibody, we distinguished four RMEC subpopulations in cultures in both media: Thy-1.1+ cells, PNA+ cells, cells negative to both reagents and cells positive to both reagents. It is likely that combined prolactin, cortisol, and insulin in CHSFM stimulate terminal differentiation of clonogenic cells.

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The therapeutic effects of WSY-0702 on benign prostatic hyperplasia in RWPE-1

  • Oh, Hyun-A;Kwon, Eun Bi;Hwang, Yun Gyeong;Park, Soon Eung;Mok, Ji Ye;Hwang, Sung Yeoun
    • CELLMED
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    • v.7 no.2
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    • pp.8.1-8.7
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    • 2017
  • Benign prostatic hyperplasia (BPH) is one of the major diseases of the urinary system in older men. WSY-0702 is the extracted from the traditional medicinal plant; Seoritae, and it has effects of anti-obesity, chronic cervical pain, and anti-oxidant. The present study aimed to investigate the therapeutic potential of WSY-0702 in the prevention and treatment of BPH. Several parameters including inflammatory mediators, hormones, and oxidative stress (OS) have been considered to play a role in the development of BPH. Prostate tissue damage and OS may lead to compensatory cellular proliferation with resulting hyperplastic growth. An in vitro study showed that proliferation inhibited the human prostate epithelial cell line RWPE-1 in a dose-dependent manner. In cell line, the cell cycle at the G2/M and G0/G1 phase and downregulated the expression of CyclineB1 (CCNB1) and CyclineD1 (CCND1). In addition, we measured the $H_2O_2$-induced OS damage using RWPE-1 cells. We examined the relative expression of protein involved in the regulation of prostate apoptosis: transforming growth factor (TGF)-${\beta}$, a negative growth factor able to induced prostate apoptosis under physiological conditions. These results suggest that WSY-0702 that can inhibit the growth of prostate epithelial cell by a mechanism that may involve arresting the cell cycle and downregulating CCNB1 and CCND1 expression. In addition, WSY-0702 exposure resulted in significant protective effects in $H_2O_2$-stressed PWPE-1 cells by reduction in TGF-${\beta}$ levels.

Visualization of mechanical stresses in expanding cell cluster (세포군집의 확장에 관여하는 물리적 힘의 가시화)

  • Cho, Youngbin;Gweon, Bomi;Ko, Ung Hyun;Shin, Jennifer H.
    • Journal of the Korean Society of Visualization
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    • v.13 no.1
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    • pp.43-48
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    • 2015
  • Collective cell migration is a fundamental phenomenon observed in various biological processes such as development, wound healing, and cancer metastasis. During the collective migration, cells undergo changes in their phenotypes from those of stable to the migratory state via the process called epithelial-mesenchymal transition (EMT). Recent findings in biology and biochemistry have shown that EMT is closely related to the cancer invasion or metastasis, but not much of the correlations in kinematics and physical forces between the neighboring cells are known yet. In this study, we aim to understand the cell migration and stress distribution within the expanding cell cluster. We constructed the in vitro cell cluster on the hydrogel, employed traction force microscopy (TFM) and monolayer stress microscopy (MSM) to visualize the physical forces within the expanding cell monolayer. During the expansion, cells at the cluster edge exhibited enhanced motility and developed focal adhesions that are the essential features of EMT while cells at the core of the cluster maintained the epithelial characteristics. In the aspect of mechanical stress, the cluster edge had the highest traction force of ~90 Pa directed toward the cluster core, which means that cells at the edge actively pull the substrate to make the cluster expansion. The cluster core of the tightly confined cells by neighboring cells had a lower traction force value (~60 Pa) but the highest intercellular normal stress of ~800 Pa because of the accumulation of traction from the edge of the monolayer.

ODONTOGENIC GHOST CELL CARCINOMA ARISING FROM ODONTOGENIC EPITHELIAL TUMOR IN MAXILLA - A CASE REPORT (치성상피종양으로부터 상악에 발생한 치성유령세포암종의 치험례)

  • Kim, Jin-Hak;Kim, Moon-Key;Cha, In-Ho;Kim, Jin;Kim, Hyun-Sil;Choi, Hee-Soo;Kim, Hyung-Jun
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.30 no.3
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    • pp.218-222
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    • 2004
  • The neoplastic variant of calcifying odontogenic cyst has various designation, and its malignant counterpart has been reported as aggressive epithelial ghost cell tumor or odontogenic ghost cell carcinoma. Odontogenic ghost cell carcinoma(OGCC) is a rare carcinoma first documented in 1985. It is composed of varying sized islands of anucleated cells with homogenous, pale eosinophilic cytoplasm, so called ghost cells, were admixed with nucleated cells. We report a case of maxillary OGCC developed from odontogenic epithelial tumor in a 25-year-old man with literature review.

Immunohistochemical analysis of the effect of low power GaAlAs laser treatment on the expression of proliferating cell nuclear antigen (PCNA) in full-thickness excisional wound of rat skin (CaAlAs 저출력 레이저 자극이 흰쥐의 피부 전층결손 절제 창상의 치유시 proliferating cell nuclear antigen(PCNA)발현에 대한 면역조직화학법적 분석)

  • Kim, Soon-Ja;Koo, Hee-Seo
    • Journal of Korean Physical Therapy Science
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    • v.10 no.1
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    • pp.198-205
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    • 2003
  • We evaluated the effect of low power GaAsAl laser on re-epithelization in full-thickness excisional wound of rat skin. Two full-thickness excisions were made on the back of the experimental animals. Low power laser applications with 10mW intensity were treated experimental animals twice a day for 7 days. On the seventh postoperative day the quantitative analysis of re-epithelization was performed using immunohistochemical staining for proliferating cell nuclear antigen (PCNA). The majority of PCNA immunoreactive cells was observed at epithelial cells in the margin of full thickness excisional wound. The low power laser treatments significantly increased the number of PCNA immunoreactive cell as compared to that of non treated animal group (p<0.01). The shape of PCNA immunoreactive cell appeared as small dark, round to ovoid structures. Most PCNA immunoreactive cells exhibited a high intensity of staining that contrasted sharply with the surrounding background. In conclusion, these findings suggest that GaAlAs laser treatments effectively enhance the epithelial wound healing by the stimulating cell proliferation. Furthermore, the majority of cell proliferation occurred in the margin of full thickness excisional wound.

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Anti-Oxidative and Anti-Inflammatory Effects of QGC in Cultured Feline Esophageal Epithelial Cells

  • Lee, Myeong Jae;Song, Hyun Ju;Jeong, Jun Yeong;Park, Sun Young;Sohn, Uy Dong
    • The Korean Journal of Physiology and Pharmacology
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    • v.17 no.1
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    • pp.81-87
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    • 2013
  • Quercetin-3-O-${\beta}$-D-glucuronopyranoside (QGC) is a flavonoid glucoside extracted from Rumex Aquaticus Herba. In the present study, anti-oxidative and anti-inflammatory effects of QGC were tested in vitro. Epithelial cells obtained from cat esophagus were cultured. When the cells were exposed to acid for 2 h, cell viability was decreased to 36%. Pretreatment with 50 ${\mu}M$ QGC for 2 h prevented the reduction in cell viability. QGC also inhibited the productions of intracellular ROS by inflammatory inducers such as acid, lipopolysaccharide, indomethacin and ethanol. QGC significantly increased the activities of superoxide dismutase (SOD) and catalase, and also induced the expression of SOD2, while it restored the decrease of catalase expression in cells exposed to acid. QGC inhibited NF-${\kappa}B$ translocation, cyclooxygenase-2 expression and $PGE_2$ secretion in cells exposed to acid, which plays an important role in the pathogenesis of esophagitis. The data suggest that QGC may well be one of the promising substances to attenuate oxidative epithelial cell injury and inflammatory signaling in esophagus inflammation.

Pathological Study on the Pulmonary Toxicity of Particulate Matters (Carbon Black, Colloidal Silica, Yellow Sands) in Mice

  • Shimada, Akinori
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2005.05a
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    • pp.51-82
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    • 2005
  • To compare the pulmonary toxicity between ultrafine colloidal silica particles (UFCSs) and fine colloidal silica particles (FCSs), mice were intratracheally instilled with 3 mg of 14-nm UFCSs and 230-nm FCSs and pathologically examined from 30 mill to 24 hr post-exposure. Histopathologically, lungs exposed to both sizes of particles showed bronchiolar degeneration and necrosis, neutrophilic inflammation in alveoli with alveolar type II cell proliferation and particle-laden alveolar macrophage accumulation. UFCSs, however, induced extensive alveolar hemorrhage compared to FCSs from 30 min onwards. UFCSs also caused more severe bronchiolar epithelial cell necrosis and neutrophil influx in alveoli than FCSs at 12 and 24 hr post-exposure. Laminin positive immunolabellings in basement membranes of bronchioles and alveoli of UFCSs treated animals was weaker than those of FCSs treated animals in all observation times. Electron microscopy demonstrated UFCSs and FCSs on bronchiolar and alveolar wall surface as well as in the cytoplasm of alveolar epithelial cells, alveolar macrophages and neutrophils. Type I alveolar epithelial cell erosion with basement membrane damage in UFCSs treated animals was more severe than those in FCSs treated animals. At 12 and 24 hr post-exposure, bronchiolar epithelia cells in UFCSs treated animals showed more intense vacuolation and necrosis compared to FCSs treated animals. These findings suggest that UFCSs has greater ability to induce lung inflammation and tissue damages than FCSs.

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Purification of Embryogenesis Stimulating Activity from Bovine Oviduct Epithelial Cell Conditioned Medium and Its Effect on the Development of Bovine Embryos (소 난관상피세포 배양액 유래 배발육촉진인자의 정제와 소 수정란 체외배양시 발육에 미치는 영향)

  • 김현일;노상호;박종임;신태영;이병천;황우석
    • Journal of Embryo Transfer
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    • v.13 no.1
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    • pp.43-52
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    • 1998
  • The embryogenesis stimulating activity(ESA) had been shown in co-culture of embryos with bovine oviduct epithelial cell(BOEC) and culture in BOEG-conditioned medium. The present study was undertaken to purify and quantify the embryotropic proteins and to determine the optimum concentration of the embryotropic protein for the proper development of embryos. In BOEC-conditioned medium, five major bands of proteins were detected(66, 53, 40, 32 and 24 kDa) by SDS-PAGE. From these proteins, 288pg of protein that had a 32kDa molecular weight was purified by gel filtration column and perfusion chromatography ion-exchange column. When purified protein was supplemented to the in vitro culture media at various concentrations in protein-free media, 2.5$\mu$g /ml supplement group showed significantly higher rates of embryo development into morula /blastocyst stages than other groups(p<0.05). In conclusion, we purified 32kDa protein from BOEC-conditioned medium and this protein showed optimum embryogenesis stimulating effect at 2.5$\mu$g /ml.

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