• Title/Summary/Keyword: biomedical laboratory

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Effect of Lipopolysaccharide (LPS) on Mouse Model of Steroid-Induced Avascular Necrosis in the Femoral Head (ANFH)

  • Ryoo, Soyoon;Lee, Sukha;Jo, Seunghyun;Lee, Siyoung;Kwak, Areum;Kim, Eunsom;Lee, Jongho;Hong, Jaewoo;Jhun, Hyunjhung;Lee, Youngmin;Sobti, Anshul Shyam;Kim, Soohyun;Oh, Kwang-Jun
    • Journal of Microbiology and Biotechnology
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    • v.24 no.3
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    • pp.394-400
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    • 2014
  • Avascular necrosis of the femoral head (ANFH) is commonly observed in patients treated with excessive glucocorticoid (GC). Single administration of lipopolysaccharide (LPS) has shown to induce immune stimulatory factors. However, the effect of repeated administration of LPS on GC-induced ANFH has not been studied. Thus, the purpose of this study was (i) to examine the cytokine profile induced by repeated LPS administrations and (ii) to test the effect of repeated LPS treatments on GC-induced ANFH. A mouse necrosis model of ANFH was designed by chronic GC administration with co-treatment of LPS. Mice body weights in the LPS/prednisolone (PDN) co-treated group were lower than that of the untreated control group, but spleen weights were greater than the control group. The levels of IL-6, $TNF{\alpha}$, and IL-33 in the liver and spleen of the LPS/PDN group were lower than the untreated control group, whereas $TNF{\alpha}$ level in the femoral head of the LPS/PDN group increased. Collectively, the effect of repeated LPS on the pathogenesis of GC-induced ANFH was associated with the $TNF{\alpha}$ level in the femoral head, but the pathogenesis did not correspond to cytokine levels in immune tissues.

Cytokine-like Activity of Liver Type Fatty Acid Binding Protein (L-FABP) Inducing Inflammatory Cytokine Interleukin-6

  • Hyunwoo Kim;Gaae Gil;Siyoung Lee;Areum Kwak;Seunghyun Jo;Ensom Kim;Tam T. Nguyen;Sinae Kim;Hyunjhung Jhun;Somi Kim;Miyeon Kim;Youngmin Lee;Soohyun Kim
    • IMMUNE NETWORK
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    • v.16 no.5
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    • pp.296-304
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    • 2016
  • It has been reported that fatty acid binding proteins (FABPs) do not act only as intracellular mediators of lipid responses but also have extracellular functions. This study aimed to investigate whether extracellular liver type (L)-FABP has a biological activity and to determined serum L-FABP levels in patients with end-stage renal disease (ESRD). We isolated L-FABP complementary deoxyribonucleic acid (cDNA) from the Huh7 human hepatocarcinoma cell line and expressed the recombinant L-FABP protein in Escherichia coli. A549 lung carcinoma and THP-1 monocytic cells were stimulated with the human recombinant L-FABP. Human whole blood cells were also treated with the human recombinant L-FABP or interleukin (IL)-1α. IL-6 levels were measured in cell culture supernatants using IL-6 enzyme-linked immunosorbent assay (ELISA). Human recombinant L-FABP induced IL-6 in a dose-dependent manner in A549, THP-1 cells, and whole blood cells. The blood samples of healthy volunteers and patients with ESRD were taken after an overnight fast. The serum levels of L-FABP in healthy volunteers and ESRD patients were quantified with L-FABP ELISA. The values of L-FABP in patients with ESRD were significantly lower than those in the control group. Our results demonstrated the biological activity of L-FABP in human cells suggesting L-FABP can be a mediator of inflammation.

Characterization of CCND1 and TWIST1 as Prognostic Markers with the Mortality Rate of Breast Cancer

  • Ahn, Sungwoo;Park, Sangjung;Wang, Hye-Young;Park, Sunyoung;Kim, Jungho;Lee, Hyeyoung
    • Biomedical Science Letters
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    • v.24 no.2
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    • pp.76-86
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    • 2018
  • Breast cancer is one of the most common cancers affecting women worldwide. Although the survival rate of breast cancer has increased, breast cancer still results in a high mortality rate. Breast cancer deaths are caused by metastasis that occurs in organ dysfunction. Recently, there have been many studies on circulating tumor cells (CTCs), which are related to breast cancer metastasis in the blood. Recent studies have demonstrated that some CTCs do not express epithelial markers. Therefore, in this study, total RNA was extracted from blood without separating out the CTCs, and the characteristics of the CTCs were analyzed by RT-qPCR. Cyclin D1 and twist-related protein 1 (TWIST1) are well-known markers for predicting the prognosis of patients with breast cancer. However, few studies have demonstrated the use of CCND1 and TWIST1 in blood as diagnostic and prognostic markers of breast cancer. In this study, patients with late-stage breast cancer had overexpressed CCND1 and TWIST1 than patients with different stages of breast cancer (P < 0.001 and P < 0.01, respectively). The relative expression level of CCND1 in survivors was higher than in patients who died (P = 0.06). The relative expression level of TWIST1 in survivors was lower than in patients who died (P = 0.08). Overall CCND1 and TWIST1 were not useful as markers for the diagnosis of breast cancer through blood. However, we showed the possibility of using CCND1 and TWIST1 as prognostic markers, and a large-scale study is needed to confirm the usefulness of these prognostic markers.

Isolation of Environmental Mycobacteria from Diverse Water Samples Using Cetylpyridinium Chloride

  • Choi, Yeon-Im;Jin, Hyun-Woo;Lee, Gyu-Sang;Kim, Jong-Bae;Song, In-Kenn;Kim, Young-Joon;Lee, Hye-Young
    • Biomedical Science Letters
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    • v.16 no.3
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    • pp.133-138
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    • 2010
  • Despite of the increasing importance of environmental mycobacteria, detection and identification of mycobacteria from environmental sources including water have been fraught with technical difficulties. Although, several protocols to optiruize isolation of mycobacteria from water sources have been reported, standard method has not yet been established. In this study, usefulness of cetylpyridinium chloride (CPC), a cationic quaternary ammonium compound, for the isolation of environmental mycobacteria from diverse water samples was evaluated. For this, water samples from diverse water sources such as effluent water, lake water, and underground water were collected, treated with diverse concentrations of CPC, and plated on the solid agar plates. Subsequently individual colonies grown on the plates were sequence analyzed for identification of each colony. In brief, the results from this study showed that the growth of mycobacteria was enhanced by use of CPC as a pre-treatment reagent to water samples by inhibiting growth of other non-mycobacteria in water. In fact, the effect of CPC to decontaminate non-mycobacteria for isolation of mycobacteria was better than 1~4% of NaOH, which is a routinely used decontaminating reagent widely employed for culturing mycobactera from sputum specimens. Therefore, the results from this study seems to support that the CPC pre-treatment may be useful for isolation of mycobacteria from diverse sources including clinical specimens which are often contaminated with other bacteria.

Detection of Mycobacterium leprae by Nested PCR Targeting M. leprae-Specific Repetitive Element (RLEP) Sequence

  • Wang, Hye-Young;Kim, Yeun;Bang, Hye-Eun;Kim, Hyun-Chul;Cho, Sang-Nae;Lee, Hye-Young
    • Biomedical Science Letters
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    • v.13 no.1
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    • pp.33-38
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    • 2007
  • The aim of this work was to validate a rapid and an accurate method for detecting Mycobacterium leprae in clinical specimens using nested PCR targeting M. leprae-specific repetitive element (RLEP) sequence. The primers were derived from the RLEP sequence which yield a 272 bp outer product and a 230 bp inner product. The specificity and the sensitivity of the nested PCR were compared with those of single PCR for detecting M. leprae using DNAs isolated from reference strain and various species of Mycobacterium. The results showed that the sensitivity of the nested PCR was about 100 to 1,000 times higher than that of the single PCR and also showed that both the single and the nested PCR were highly specific to M. leprae. Subsequently, the usefulness of the single and nested PCR was evaluated with clinical samples isolated from leprosy patients. The number of positive detections by the single and the nested PCR with a total of 20 specimens from leprosy patients were 9 (45%) and 20 (100%), respectively. The results clearly showed that nested PCR has highest sensitivity in detecting M. leprae from clinical specimens. Therefore, nested primers targeting RLEP sequence developed in this study seems to be useful to detect the presence of M. leprae.

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Gingko biloba Extract Ameliorates Colonic Inflammation in DSS-induced Model of Colitis in Mice

  • Rhee, Ki-Jong;Gwon, Sun-Yeong;Hwang, Soonjae;Lee, Chang Gun;Jang, In-Ho;Wie, Myung-Bok;Jung, Bae Dong
    • Biomedical Science Letters
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    • v.20 no.4
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    • pp.227-236
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    • 2014
  • Ulcerative colitis (UC) is a serious gastrointestinal tract disease characterized by recurrent chronic inflammation and mucosal damage of the gastrointestinal tract. The conventional therapies of choice are anti-inflammatory agents, steroids and anti-TNF-${\alpha}$ therapy. However, inherent limitations in these therapies have steered many UC patients to supplement existing therapies with alternative medicinal products. In the current study, we tested the efficacy of Gingko bilola extract (EGb 761) in abating colonic inflammation in a DSS-induced murine model of colitis. C57BL/6 mice were administered 2% DSS in the drinking water for 7 days, then regular water for 7 days, and then 2% DSS for an additional 7 days. EGb 761 (1 mg/dose) was oral gavaged daily for the duration of the experiment. At the termination of the experiment, mice treated with EGb+DSS showed higher body weight, lower spleen weight and longer colon length compared to mice treated with DSS alone. HE-stained colon tissues also exhibited less histologic inflammation in mice treated with EGb+DSS mice compared to mice treated with DSS alone. The serum levels inflammatory cytokines, KC and TNF-${\alpha}$, were also decreased in mice treated with EGb+DSS compared to mice treated with DSS alone. Finally, addition of EGb 761 to TNF-${\alpha}$ treated colonic cell line (HT29/c1) decreased secretion of IL-8 in vitro. These results collectively suggest that EGb 761 abates induction of colitis in DSS-induced model of colitis in mice.

Immunostimulating Effects of Polysaccharides of Panax ginseng by Oral Administration into BALB/c Mice

  • Kang, Mi-Hyeon;Park, A-Young;Yang, Ae-Hee;Park, Ye-Eun;Hong, Seung-Hye;Park, Gab-Soon;Jung, Suk-Yul
    • Biomedical Science Letters
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    • v.20 no.4
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    • pp.194-200
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    • 2014
  • Ginsan, polysaccharide isolated from the root of Panax ginseng C.A. Meyer, has been shown to be a potent immunomodulator, producing several cytokines and stimulating lymphoid cells to proliferate. In this study, ginsan was orally inoculated into BALB/c mice up to 39 days and the activity of immune cells containing macrophages and T cells was analyzed. Moreover, the production of cytokines, e.g., tumor necrotic factor-${\alpha}$ (TNF-${\alpha}$), interferon-${\gamma}$ (IFN-${\gamma}$), GM-CSF and IL-12 was also analyzed. In results, the phagocytosis of macrophages was increased. About 13% cytotoxicity of NK cells was observed in 22 days and 29 days of administration. But, oral administration did not highly affect the proliferation of T cells. In cytokine analysis, 150 mg/kg and 300 mg/kg at 22 days and 29 days showed three times more increase in TNF-${\alpha}$ than the controls. IFN-${\gamma}$ showed 1.07 and 1.16 times more increase at 150 mg/kg and 300 mg/kg over 22 days, respectively more than the controls. 32 days of 150 mg/kg and 300 mg/kg induced GM-CSF of about 1.3 times more than the controls. IL-12 was not induced in samples more than the controls. Ginsan could be a potential immunostimulator. Therefore, our study suggests that it can be adapted as an immunostimulator that requires a relatively short oral administration.

Assessment of Ki-67 for Predicting Effective Prognosis in Breast Cancer Subtypes

  • Park, Sangjung;Park, Sunyoung;Kim, Jungho;Ahn, Sungwoo;Park, Kwang Hwa;Lee, Hyeyoung
    • Biomedical Science Letters
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    • v.24 no.1
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    • pp.9-14
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    • 2018
  • Ki-67 has been widely performed and become an important biomarker in worldwide clinics, but the standard cut off value of Ki-67 index in breast cancer is still controversy. The objective study was to understand the Ki-67 in breast cancer subtypes and to investigate relative risk of breast cancer subtypes according to Ki-67 cut off value in Korean breast cancer. Immunohistochemical staining (IHC) for estrogen receptor (ER), progesterone receptor (PR), human epidermal growth factor receptor 2 (HER2), and Ki-67 index was examined from 123 breast cancer patients. Ki-67 index was significantly overexpressed in PR, ER, and HER2 hormone negative groups. Ki-67 index in Triple negative and HER2 subtypes was shown significantly higher than that in Luminal A and Luminal B subtype. Then, we compared the relative risk of each subtype according to 14% and 20% Ki-67 cut off value, which were applied in most clinics. Especially, 20% Ki-67 cut off value in HER2 and Triple negative subtypes was shown 8.41 fold and 2.83 fold higher relative risk than this in Luminal A subtype. Moreover, Ki-67 index in HER2 2+ or 3+ status showed significantly overexpressed than this in HER2 1+ status. At the 20% Ki-67 cut off value, HER2 1+ or 2+ status and 3+ status showed significant difference. Therefore, the 20% Ki-67 cut off value will be useful as a precise prognostic management and helpful for interpreting diverse outcomes of other subtypes in breast cancer patients.

Identification of Proteins Interacting with C- Terminal Region of Human Ankyrin-G

  • Lee, Yeong-Mi;Lee, Min-A;Park, Jae-Kyoung;Kim, Myong-Shin;Jeon, Eun-Bee;Park, Su-Il;Kim, Chong-Rak
    • Biomedical Science Letters
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    • v.9 no.3
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    • pp.159-165
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    • 2003
  • Ankyrins are a ubiquitously expressed family of intracellular adaptor proteins involved in targeting diverse proteins to specialized membrane domains in both the plasma membrane and the endoplasmic reticulum. Recently, the studies with C-terminus of ankyrins have identified that ankyrin-B is capable of interacting with Hsp40 and sAnkl is capable of interacting with obscurin and titin, but the function of C-terminal domain of ankyrin-G remains unknown. To identify proteins interacting C-terminus of ankyrin-G, we used the C-terminus of ankyrin-G as a bait for a yeast two-hybrid screen of brain cDNA library. Approximately 1.33$\times$l0$^6$ transformants were screened, of which 13 positive clones were obtained as determined by activation of HIS3, ADE2 and MELl reporter genes. Sequence analyses of these 13 plasmids revealed that cDNA inserts of 13 colonies showed highly homologous to 11 genes, including 5 known (i.e., Na$^+$/K$^+$ ATPase $\beta$1, SERBPl, UTF2, cytochrome C oxidase and collagen IV $\alpha$2) and 6 unknown genes. The evaluation of the proteins that emerge from these experiments provides a rational approach to investigate the those proteins significant in interaction with ankyrin-G.

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