• Title/Summary/Keyword: Clone cells

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A case of transitional carcinoma in the nasal cavity of a dog (개의 비강 내 이행암종 증례)

  • Kang, Hwa-Jung;Kang, Sang-Chul;Jung, Ji-Youl;Jo, Suk-Hee;Roh, In-Soon;Lee, Joo-Myung;Cheong, Jongtae;Kim, Jung-Hun;An, Min-Chan;Bae, Jong-Hee;Kim, Jae-Hoon
    • Korean Journal of Veterinary Research
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    • v.48 no.3
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    • pp.305-310
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    • 2008
  • A 10-year-old female Yorkshire terrier with the clinical signs of nasal swelling, epistaxis and nasal discharge was presented to the Veterinary Teaching Hospital in the Cheju National University. Abnormal nasal mass was detected in physical examination and radiographic findings. After surgical excision, the sample of nasal mass was referred to Pathology Department of Veterinary Medicine. Grossly, the mass was soft, friable, and $2.5{\times}4cm$ cm in size. Histopathologically, the mass was composed of mediumsized non-keratinizing columnar to polyhedral cells arranged in anastomosing ribbon and large nest. It has complex in-folding of thick epithelial layers separated by fibrovascular septa. Tumor cells showed characteristic palisading arrangement of columnar cells, and perpendicularly distributed to the basement membrane. The cells had pale basophilic cytoplasm, oval nucleus and one or more nucleoli, and indistinct cellular border. Many tumor cell emboli were presented in lymphatics. Immunohistochemistry revealed that tumor cells were cytokeratin (CK) 19 and CK clone MNF116 positive and but CK7 and CK high molecular weight negative. Based on the gross, histopathologic, and immunohistochemical findings, the mass was diagnosed as transitional carcinoma in nasal cavity. In our best knowledge, this is the first report of transitional carcinoma originated from transitional zone of canine nasal cavity in Korea.

Cross-reactivity of Human Polyclonal Anti-GLUT1 Antisera with the Endogenous Insect Cell Glucose Transporters and the Baculovirus-expressed GLUT1

  • Lee, Chong-Kee
    • Biomedical Science Letters
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    • v.7 no.4
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    • pp.161-166
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    • 2001
  • Most mammalian cells take up glucose by passive transport proteins in the plasma membranes. The best known of these proteins is the human erythrocyte glucose transporter, GLUT1. High levels of heterologous expression far the transporter are necessary for the investigation of its three-dimensional structure by crystallization. To achieve this, the baculovirus expression system has become popular choice. However, Spodoptera frugiperda Clone 9 (Sf9) cells, which are commonly employed as the host permissive cell line to support baculovirus replication and protein synthesis, grow well on TC-100 medium that contains 0.1% D-glucose as the major carbon source, suggesting the presence of endogenous glucose transporters. Furthermore, very little is known of the endogenous transporters properties of Sf9 cells. Therefore, human GLUT1 antibodies would play an important role for characterization of the GLUT1 expressed in insect cell. However, the successful use of such antibodies for characterization of GLUT1 expression m insect cells relies upon their specificity for the human protein and lack of cross-reaction with endogenous transporters. It is therefore important to determine the potential cross-reactivity of the antibodies with the endogenous insect cell glucose transporters. In the present study, the potential cross-reactivity of the human GLUT1 antibodies with the endogenous insect cell glucose transporters was examined by Western blotting. Neither the antibodies against intact GLUT1 nor those against the C-terminus labelled any band migrating in the region expected fur a protein of M$_r$ comparable to GLUT1, whereas these antibodies specifically recognized the human GLUT1. Specificity of the human GLUT1 antibodies tested was also shown by cross-reaction with the GLUT1 expressed in insect cells. In addition, the insect cell glucose transporter was found to have very low affinity for cytochalasin B, a potent inhibitor of human erythrocyte glucose transporter.

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Ginsenoside compound K inhibits nuclear factor-kappa B by targeting Annexin A2

  • Wang, Yu-Shi;Zhu, Hongyan;Li, He;Li, Yang;Zhao, Bing;Jin, Ying-Hua
    • Journal of Ginseng Research
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    • v.43 no.3
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    • pp.452-459
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    • 2019
  • Background: Ginsenoside compound K(C-K), a major metabolite of ginsenoside, exhibits anticancer activity in various cancer cells and animal models. A cell signaling study has shown that C-K inhibited nuclear factor-kappa B ($NF-{\kappa}B$) pathway in human astroglial cells and liver cancer cells. However, the molecular targets of C-K and the initiating events were not elucidated. Methods: Interaction between C-K and Annexin A2 was determined by molecular docking and thermal shift assay. HepG2 cells were treated with C-K, followed by a luciferase reporter assay for $NF-{\kappa}B$, immunofluorescence imaging for the subcellular localization of Annexin A2 and $NF-{\kappa}B$ p50 subunit, coimmunoprecipitation of Annexin A2 and $NF-{\kappa}B$ p50 subunit, and both cell viability assay and plate clone formation assay to determine the cell viability. Results: Both molecular docking and thermal shift assay positively confirmed the interaction between Annexin A2 and C-K. This interaction prevented the interaction between Annexin A2 and $NF-{\kappa}B$ p50 subunit and their nuclear colocalization, which attenuated the activation of $NF-{\kappa}B$ and the expression of its downstream genes, followed by the activation of caspase 9 and 3. In addition, the overexpression of Annexin A2-K320A, a C-K binding-deficient mutant of Annexin A2, rendered cells to resist C-K treatment, indicating that C-K exerts its cytotoxic activity mainly by targeting Annexin A2. Conclusion: This study for the first time revealed a cellular target of C-K and the molecular mechanism for its anticancer activity.

Zebrafish (Danio rerio) Thyroid Hormone Receptor $\alpha$1 Counteracts Retinoic Acid-induced Transcription

  • Rhee, Myubg-Chull;Lee, Woonghee;Chang, Mi-Sook;Lee, Sang-Kyou
    • Animal cells and systems
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    • v.2 no.1
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    • pp.133-137
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    • 1998
  • The present study aims to characterize a cDNA encoding zebrafish thyroid hormone receptor $\alpha{1}$ $(zTR\alpha{1)}$ in order to investigate its possible role in the early stage of embryonic development. A mobility shift assay showed that $zTR\alpha{1}$ overexpressed in COS7 cells specifically bound to thyroid hormone response element (TRE). In addition, the specific interaction of anti-rat $TR\alpha{1}$ antibodies with $zTR\alpha1$/TRE complexes demonstrated that the cDNA clone encoded zebrafish thyroid hormone receptor $\alpha{1}$. Transient cotransfection assays showed that $zTR\alpha{1}$ repressed the transcription which was induced by retinoic acid (RA), a well-characterized embryonic morphogen. These results suggest that zTRal may be involved in regulating the RA-induced gene transcription during early embryonic development.

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Significance of Some Proliferation Markers and Some Prognostic Factors in Patients with Multiple Myeloma and their Impact on the Patients' Survival

  • Abdelgawad, Iman A.;Radwan, Noha H.;Shafik, Roxan E.;Shokralla, Hala A.
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.5
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    • pp.2389-2394
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    • 2016
  • Background: Multiple myeloma (MM) is influenced by genetic and micro-environmental changes. Malignant plasma cells produce an abnormal monoclonal immunoglobulin, as well as cytokines, such as IL-10 and IL-6 which stimulate cells of the bone marrow microenvironment (BMM) and cause dysfunction and failure of many organs. B cell activating factor (BAFF), IL6, IL10 are known to influence the growth & survival of the malignant clone. Aim: The objectives of the present study were to investigate the circulating levels of BAFF, IL-10 and IL-6, correlate them with well-known parameters of disease activity in patients with MM, and to detect their impact on the patients' survival. Materials and Methods: This study was conducted on 89 newly diagnosed MM patients and seventy apparently healthy volunteers as a normal control group. BAFF, IL6, IL10 were measured by ELISA for both groups. Survival analysis was performed for all patients. Results: Studied markers were higher in the MM patients compared to the normal control subjects. Patients' survival was improved by high serum BAFF levels. Conclusions: High levels of BAFF were found to improve patients' survival. BAFF and IL-6 can be considered probable diagnostic markers for MM.

Expression and Characterization of the Human Immunodeficiency Virus Type 1 Mutant Envelope Glycoproteins in Mammalian Cells (진핵세포에서 HSV-1 Envelope 변이 단백질의 발현 및 발현 단백질의 특성 연구)

  • Ryu, Ji-Yoon;Park, Jin-Seu
    • The Journal of Korean Society of Virology
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    • v.29 no.3
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    • pp.183-193
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    • 1999
  • Human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein is synthesized as a 160 KDa precursor, gp160, that is cleaved by a cellular protease to form the gp120 and gp41 subunits. Mammalian expression vectors were designed that are capable of efficient expression of various mutant envelope glycoproteins derived from a molecular clone of HIV-1. To construct these vectors, one type of mutation was made at the gp120-gp41 cleavage site by oligonucleotide-directed mutagenesis. And another mutation was made to change amino acids in the membrane spanning region of HIV-1 gp41 important for membrane anchorage. Next, these two mutations were combined to generate a vector to have double mutations in cleavage site and membrane-spanning region. These mutants were transiently expressed in mammalian cells. The effect of these mutations on envelope glycoprotein synthesis, proteolytic processing and secretion was determined. In addition, cell surface expression and ability of the glycoprotein to induce syncytium formation were examined. This study provides a mammalian expression system that is capable of efficient expression and secretion of soluble gp160.

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Molecular cloning and expression of glyceraldehyde-3-phosphate dehydrogenase gene under environmental stresses in sweetpotato

  • Kim, Young-Hwa;Song, Young-Sun;Huh, Gyung-Hye
    • Journal of Plant Biotechnology
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    • v.35 no.2
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    • pp.95-100
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    • 2008
  • Glyceraldehyde-3-phosphate dehydrogenase (GAPDH), a main enzyme in the glycolytic pathway, is involved in cellular energy production and regarded as a housekeeping gene. Previously, cytosolic GAPDH was selected as the most significantly abundant gene in EST library of sweetpotato suspension cells. In this study, a full-length of cDNA clone (IbGAPDH) encoding GAPDH was isolated from suspension-cultured cells of sweetpotato (Ipomoea babatas), and its expression was investigated with a view to understanding the physiological function of GAPDH in relation to environmental stresses. IbGAPDH encoded a 36.9 kDa polypeptide consisting of 337 amino acids. When the deduced amino acid of IbGAPDH was compared with other higher plants, IbGAPDH showed high homology with cytosolic GAPDH. The mRNA level of IbGAPDH significantly increased under environmental stresses, such as $H_2O_2$, MV and cold treatments. Among them, the transcript level of IbGAPDH gene was the highest under cold stress. Further investigation of the transcription level under $10^{\circ}C$ or $15^{\circ}C$ was performed with different tissues of sweetpotato. The transcription of IbGAPDH was increased by cold stress with tissue-specificity, moreover, showed different patterns according to temperature.

A Study on the Physiological Effects and Dyeing Properties of the Extract of Fermented Indigo(Part II) (발효쪽 추출물의 생리적 기능과 염색특성(제2보))

  • 한신영;최석철
    • Journal of the Korean Society of Clothing and Textiles
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    • v.24 no.8
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    • pp.1146-1154
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    • 2000
  • The purpose of this study was to investigate the antimicrobial activity, antimutagenic and anticancer effects and dyeing properties of the fermented indigo extract. The methanol extract of fermented indigo showed a strong inhibition effect on Trich. mentagrophytes and antimutagenic activities against aflatoxin B$_1$(AFB$_1$) in the Ames test using Salmonella typhimurium TA 100. We also found in vitro anticancer effects of the methanol extract of fermented indigo and it was evaluated by using Clone M-3 mouse melanoma cells and A431 human epidermoid carcinoma cells and exerted little cytotoxity against 3T3-L1 embryo fibroblast cell. In the relationship between the K/S values of cotton and silk fabrics dyed with fermented indigo and dyeing repeating times, the K/S values became higher as the repeating times were increased. The K/S values were high when the fabrics were dyed at low temperture. The K/S values of cotton fabrics were higher than those of silk fabrics. Changes of surface color of silk fabrics were higher than those of cotton fabrics after water fastness test, laundering, irradiation and treatment of acidic and alkaline perspiration.

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Production of Monoclonal Antibodies by Hybridomas Sensitized to Sporozoites of Cryptosporidium parvum (Cryptosporidium parvum Sporozoites 에 감작된 Hybridomas 에서의 Monoclonal Antibody 생산)

  • Cho, Myung-Hwan
    • Microbiology and Biotechnology Letters
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    • v.17 no.5
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    • pp.494-498
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    • 1989
  • Hybridoma cell lines, which secrete monoclonal antibodies (mAbs) against the surface antigens of Cryptosporidium parvum Sporozoites, were produced by fusing spleen cells of C. parvum Sporozoite-immunized mice with P3-X63-Ag8 myeloma cells. Two cloned antibody-secreting cell lines, Kor1 and Ea2, were established and produced IgG1 and IgG2a antibodies, respectively. Percoll-purified sporozoites were solubilized and separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Western blot assay demonstrates that an antigen of 20-kDa was bound by monoclonals. By indirect immunofluorescence microscopy, mAb exhibited uniform binding to the sporozoite surface.

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Strain Improvement of Yeast for Ethanol Production Using a Combined Treatment of Electric Field and Chemical Mutagen N-Methyl-N'-nitro-N- nitrosoguanidine

  • Kim, Keun;Lee, Jae-Yeon
    • Journal of Microbiology and Biotechnology
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    • v.8 no.2
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    • pp.119-123
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    • 1998
  • The feasibility of using combined treatments of electric field and chemical mutagen N-methyl-N'-nitro-N-nitroso-guanidine (NTG) for the strain improvement of Saccharomyces sp. in ethanol production was examined. The treatment of electric field alone resulted in no effect on the lethality of yeast cells under the conditions of this study. However, when the electric field was applied together to the treatment of yeast cells with NTG, the electric field increased the lethal effect and auxotrophic mutation rate of NTG. The combined treatment of electric field and NTG also increased the chances of. obtaining superior yeast strains for the ethanol production from tapioca. A higher number of improved clones was obtained by the combined treatments of electric field and NTG than by the NTG treatment alone. The best clone, NF 30-9, which was also obtained by the combined treatment, produced $11.07\%$ (w/v) ethanol from tapioca slurry containing 25% (w/v) reducing sugar while the parental strain produced 9.77% (w/v).

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