• Title/Summary/Keyword: WB cell

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Inhibition of Gap Junctional Intercellular Communication by Food Preservatives Potassium Sorbate (소르빈산 칼륨의 GJIC 억제로 인한 간독성 유발)

  • Hwang, Jae-Woong;Chung, Ji-Hye;Jung, Ji-Won;Jung, Ji-Youn;Kim, Sun-Jung;Park, Jung-Ran;Ahn, Ji-Yun;Ha, Tae-Youl;Kim, Sung-Ran;Lee, Yong-Soon;Kang, Kyung-Sun
    • Journal of Food Hygiene and Safety
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    • v.21 no.4
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    • pp.269-273
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    • 2006
  • Potassium Sorbate (PS) is a potassium salt version of sorbic acid, which has antimicrobial and fungistatic features in foods. Therefore, PS is used as a food preservative against fungi and mold. PS has been found to be non-toxic even when taken in large quantities given its trait to be broken down in the body into water and carbon dioxide. Gap Junctional Intercellular Communication (GJIC) is essential in the maintenance of tissue homeostasis during development and differentiation. This study was made of the effects of PS on GJIC in WB-F344 rat liver epithelial (WB) cells. We found dramatic decrease of cell viability in time- and dose-dependent manners when WB cells were treated with PS. The effect of PS on GJIC is strong inhibition, and it took place in parallel with a hyperphosphorylation of connexin 43 expression. The finding that PS interferes with gap junction functionality should be considered with respect to the mechanism of PS-induced hepatotoxicity.

Effects of Pahs and Pcbs and Their Toxic Metabolites on Inhibition of Gjic and Cell Proliferation in Rat Liver Epithelial Wb-F344 Cells

  • Miroslav, Machala;Jan, Vondracek;Katerina, Chramostova;Lenka, Sindlerova;Pavel, Krcmar;Martina, Pliskova;Katerina, Pencikova;Brad, Upham
    • Environmental Mutagens and Carcinogens
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    • v.23 no.2
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    • pp.56-62
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    • 2003
  • The liver progenitor cells could form a potential target cell population fore both tumor-initiating and -promoting chemicals. Induction of drug-metabolizing and antioxidant enzymes, including AhR-dependent CYP1A1, NQO-1 and AKR1C9, was detected in the rat liver epithelial WB-F344 "stem-like" cells. Additionally, WB-F344 cells express a functional, wild-type form of p53 protein, a biomarker of genotoxic events, and connexin 43, a basic structural unit of gap junctions forming an important type of intercellular communication. In this cellular model, two complementary assays have been established for detection of the modes of action associated with tumor promotion: inhibition of gap junctional intercellular communication (GJIC) and proliferative activity in confluent cells. We found that the PAHs and PCBs, which are AhR agonists, released WB-F344 cells from contact inhibition, increasing both DNA synthesis and cell numbers. Genotoxic effects of some PAHs that lead to apoptosis and cell cycle delay might interfere with the proliferative activity of PAHs. Contrary to that, the nongenotoxic low-molecular-weight PAHs and non-dioxin-like PCB congeners, abundant in the environment, did not significantly affect cell cycle and cell proliferation; however both groups of compounds inhibited GJIC in WB-F344 cells. The release from contact inhibiton by a mechanism that possibly involves the AhR activation, inhibition of GJIC and genotoxic events induced by environmental contaminants are three important modes of action that could play an important role in carcinogenic effects of toxic compounds. The relative potencies to inhibit GJIC, to induce AhR-mediated activity, and to release cells from contact inhibition were determined for a large series of PAHs and PCBs and their metabolites. In vitro bioassays based on detection of events on cellular level (deregulation of GJIC and/or proliferation) or determination of receptor-mediated activities in both ?$stem-like^{\circ}{\times}$ and hepatocyte-like liver cellular models are valuable tools for detection of modes of action of polyaromatic hydrocarbons. They may serve, together with concentration data, as a first step in their risk assessment.

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Gene Expression Analysis of Rat Liver Epithelial Cells in Response to Thioacetamide

  • Park, Joon-Suk;Yeom, Hye-Jung;Jung, Jin-Wook;Hwang, Seung-Yong;Lee, Yong-Soon;Kang, Kyung-Sun
    • Molecular & Cellular Toxicology
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    • v.1 no.3
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    • pp.203-208
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    • 2005
  • Thioacetamide (TA) is potent haptotoxincant that requires metabolic activation by mixed-function oxidases. Micrcarray technology, which is massive parallel gene expression profiling in a single hybridization experiment, has provided as a powerful molecular genetic tool for biological system related toxicant. In this study we focus on the use of toxicogenomics for the determination of gene expression analysis associated with hepatotoxicity in rat liver epithelial cell line WB-F344 (WB). The WB cells was used to assess the toxic effects of TA. WB cells were exposed to two concentrations of TA-doses which caused 20% and 50% cell death were chosen and the cells exposed for periods of 2 and 24 h. Our data revealed that following the 2-h exposure at the both of doses and 24-h exposure at the low doses, few changes in gene expression were detected. However, after 24-h exposure of the cells to the high concentration, multiple changes in gene expression were observed. TA treatment gave rise predominantly to up-regulation of genes involved in cell cycle and cell death, but down-regulation of genes involves in cell adhesion and calcium ion binding. Exposure of WB cells to higher doses of the TA gave rise to more changes in gene expression at lower exposure times. These results show that TA regulates expression of numerous genes via direct molecular signaling mechanisms in liver cells.

Comparison of growth performance and related gene expression of muscle and fat from Landrace, Yorkshire, and Duroc and Woori black pigs

  • Bosung Kim;Yejin Min;Yongdae Jeong;Sivasubramanian Ramani;Hyewon Lim;Yeonsu Jo;Woosang Kim;Yohan Choi;Sungkwon Park
    • Journal of Animal Science and Technology
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    • v.65 no.1
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    • pp.160-174
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    • 2023
  • The purpose of this study was to compare marbling score, meat quality, juiciness, sarcomere length, and skeletal muscle satellite cell (SMSC) growth and related gene expression between Woori black pig (WB) and the Landrace, Yorkshire, and Duroc (LYD) crossbreed at different body weights (b.w.). WB was developed to improve meat quality and growth efficiency by crossbreeding Duroc with Korean native black pig. A total of 24 pigs were sacrificed when their b.w. reached about 50, 75, 100, and 120 kg. SMSC were isolated from the femoris muscles, and muscle and adipose tissues were sampled from the middle and the subcutaneous part of the femoris of hind legs, respectively. Expression levels of genes including Myoblast determination protein 1 (MyoD), Paired box gene 3 (Pax3), Myosin heavy chain (MyHC), and Myogenin, which are responsible for the growth and development of SMSC, were higher in LYD than the WB. Muscle growth inhibitor myostatin (MSTN), however, was expressed more in WB compared to LYD (p < 0.01). Numbers of SMSC extracted from femoris muscle of LYD at 50, 75, 100, and 120 kg b.w. were 8.5 ± 0.223, 8.6 ± 0.245, 7.2 ± 0.249, and 10.9 ± 0.795, and those from WB were 6.2 ± 0.32, 6.2 ± 0.374, 5.3 ± 0.423, and 17.1 ± 0.315, respectively. Expression of adipogenic genes in adipose tissue including CCAAT/enhancer-binding protein (CEBP)-β, peroxisome proliferator activated receptor (PPAR)-γ, and fatty acid synthase (FASN), were greater in WB when compared with LYD (p < 0.01). Results from the current study suggest that different muscle cell numbers between 2 different breeds might be affected by related gene expression and this warrants further investigation on other growth factors regulating animal growth and development.

Expression of a Bacillus subtilis Endoglucanase in Protease-Deficient Bacillus subtilis Strains

  • Yang, Mi-Jeong;Jung, Sun-Hwa;Shin, Eun-Sun;Kim, Jung-Ho;Yun, Han-Dae;Wong, Sui-Lam;Kim, Ho-On
    • Journal of Microbiology and Biotechnology
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    • v.14 no.2
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    • pp.430-434
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    • 2004
  • Three extracellular protease-deficient Bacillus subtilis strains were transformed with the plasmid pCK98 containing the endo-$\beta$-1,4-glucanase (Eng) gene of B. subtilis BSE616. The three transformants, B. subtilis DB104 (pCK98), WB600 (pCK98) and WB700 (pCK98), produced the same high level of enzyme activity and showed similar patterns of cell growth and enzyme production. When B. subtilis DB 104 (pCK98), a two-extracellular protease deficient strain, was cultured for 22 h, almost all the secreted enzyme was found to be in the completely cleaved form by both activity staining and Western blotting studies. B. subtilis WB600 (pCK98), a six-extracellular protease-deficient strain, produced a partially cleaved form in addition to the intact form of the enzyme, although the degree of internal cleavage of the enzyme was greatly reduced. With B. subtilis WB700 (pCK98), a seven-extracellular protease-deficient strain, almost all the enzyme was produced as the intact uncleaved form. This study illustrates that a role of the V pr protease is to degrade foreign proteins produced in B. subtilis and WB700 is a suitable expression system for producing the intact form of the Eng and other foreign proteins that may lose at least part of their efficacy due to internal proteolytic cleavage.

Propranolol Inhibits the Proliferation of Human Glioblastoma Cell Lines through Notch1 and Hes1 Signaling System

  • Kim, Hyun Sik;Park, Young Han;Lee, Heui Seung;Kwon, Mi Jung;Song, Joon Ho;Chang, In Bok
    • Journal of Korean Neurosurgical Society
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    • v.64 no.5
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    • pp.716-725
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    • 2021
  • Objective : The anti-tumor effect of the beta-adrenergic receptor antagonist propranolol in breast cancer is well known; however, its activity in glioblastoma is not well-evaluated. The Notch-Hes pathway is known to regulate cell differentiation, proliferation, and apoptosis. We investigated the effect of propranolol to human glioblastoma cell lines, and the role of Notch and Hes signaling in this process. Methods : We performed immunohistochemical staining on 31 surgically resected primary human glioblastoma tissues. We also used glioblastoma cell lines of U87-MG, LN229, and neuroblastoma cell line of SH-SY5Y in this study. The effect of propranolol and isoproterenol on cell proliferation was evaluated using the MTT assay (absorbance 570 nm). The impact of propranolol on gene expression (Notch and Hes) was evaluated using real-time polymerase chain reaction (RT-PCR, whereas protein levels of Notch1 and Hes1 were measured using Western blotting (WB), simultaneously. Small interfering RNA (siRNA) was used to suppress the Notch gene to investigate its role in the proliferation of glioblastoma. Results : Propranolol and isoproterenol caused a dose-dependent decrease in cell proliferation (MTT assay). RT-PCR showed an increase in Notch1 and Hes1 expression by propranolol, whereas WB demonstrated increase in Notch1 protein, but a decrease in Hes1 by propranolol. The proliferation of U87-MG and LN229 was not significantly suppressed after transfection with Notch siRNA. Conclusion : These results demonstrated that propranolol suppressed the proliferation of glioblastoma cell lines and neuroblastoma cell line, and Hes1 was more closely involved than Notch1 was in glioblastoma proliferation.

Media Optimization and Comparison of Fermentation Type for Overproduction of Staphylodinase in Bacillus subtilis WB700 (Bacillus Subtilis W700에서의 Staphylpkinase 대량생산을 위한 배지 최적화 및 배양방법의 비교)

  • 박인석;김병기
    • KSBB Journal
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    • v.16 no.4
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    • pp.415-419
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    • 2001
  • To produce staphylokinase (SAK) in B. subtilis WB700, media optimization was carried out and the operation of batch and fed-batch fermentation were compared. Tryptone is a good nitrogen source and its optimum concentration in modified super rich(MSR) media is 15 g/L. When glucose is used as a limiting carbon source in the MSR media, 5 g/L of an optimum glucose concentration was identified for the SAK production under the control of P43 promoter. As the expression of P43 promoter is controlled by the limitation of oxygen, the SAK production was controlled at the 30% DO level in the fed-batch fermentation. Unexpectedly, batch fermentation using MSR media showed 1.5 times higher yield of SAK than that of the fed-batch fermentation. The main cause of the results comes from not achieving higher cell concentration in the fed-batch fermentation and the optimum expression level of P43 promoter under oxygen or nutrient limitations. We could not achieve the increase in cell concentration by any means in batch culture as well as fed-batch culture. The highest yield in the batch culture was 2880 units of SAK activity and 455 mg/L of secreted SAK.

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Inhibition of Gap Junctional Intercellular Communication in Rat Liver Epithelial Cells Induced by BHT and Propyl Gallate (간상피세포에서 BHT와 propyl gallate에 의한 gap junctional intercellular communication 억제 효과)

  • Kim, Ji-Sun;Kim, Sung-Ran;Ahn, Ji-Yun;Ha, Tae-Youl;Kang, Kyoung-Sun;Kim, Sun-A
    • Korean Journal of Food Science and Technology
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    • v.39 no.5
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    • pp.558-563
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    • 2007
  • This study was conducted to analyze the cytotoxic effects of butylated hydroxytoluene (BHT) and propyl gallate (PG) in WB-F344 rat liver epithelial cells. Here we measured the inhibition level of gap junctional intercellular communication (GJIC) and elucidated the relationships between GJIC and mitogen-activated protein kinases (MAPKs) such as ERK, JNK, and p38. The cytotoxicities of BHT and PG appeared at concentrations of 1.0mM and 0.25mM, respectively, in the WB-F344 cells; and GJIC inhibition, which was analyzed by a scrape-loading/dye transfer assay and Western blotting analysis, appeared at 0.6mM for BHT and 0.1mM for PG, respectively. Also, the phosphorylations of Cx43, ERK, JNK, and p38 increased in dose-dependent manners. This suggests that BHT and PG treatments inhibited GJIC by the phosphorylation of MAPKs prior to cell damage.

Diagnostic Performance of Whole-Body Diffusion-Weighted Imaging Compared to PET-CT Plus Brain MRI in Staging Clinically Resectable Lung Cancer

  • Usuda, Katsuo;Sagawa, Motoyasu;Maeda, Sumiko;Motono, Nozomu;Tanaka, Makoto;Machida, Yuichiro;Matoba, Takuma Matsui Munetaka;Watanabe, Naoto;Tonami, Hisao;Ueda, Yoshimichi;Uramoto, Hidetaka
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.6
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    • pp.2775-2780
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    • 2016
  • Background: Precise staging of lung cancer is usually evaluated by PET-CT and brain MRI. Recently, however, whole-body diffusion-weighted magnetic resonance imaging (WB-DWI) has be applied. The aim of this study is to determine whether the diagnostic performance of lung cancer staging by WB-DWI is superior to that of PET-CT+brain MRI. Materials and Methods: PET-CT + brain MRI and WB-DWI were used for lung cancer staging before surgery with 59 adenocarcinomas, 16 squamous cell carcinomas and 6 other carcinomas. Results: PET-CT + brain MRI correctly identified the pathologic N staging in 67 patients (82.7%), with overstaging in 5 (6.2%) and understaging in 9 (11.1%), giving a staging accuracy of 0.827. WB-DWI correctly identified the pathologic N staging in 72 patients (88.9%), with overstaging in 1 (1.2%) and understaging in 8 patients (9.9%), giving a staging accuracy of 0.889. There were no significant differences in accuracies. PET-CT + brain MRI correctly identified the pathologic stages in 56 patients (69.1%), with overstaging in 7 (8.6%) and understaging in 18 (22.2%), giving a staging accuracy of 0.691. WB-DWI correctly identified the pathologic stages in 61 patients (75.3%), with overstaging in 4 (4.9%) and understagings in16(19.7%), giving a staging accuracy of 0.753. There were no significant difference in accuracies. Conclusions: Diagnostic efficacy of WB-DWI for lung cancer staging is equivalent to that of PET-CT + brain MRI.