• Title/Summary/Keyword: Flow cytometric analysis

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Eudesmols Induce Apoptosis through Release of Cytochrome c in HL-60 Cells

  • Hoang, Duc Manh;Trung, Trinh Nam;He, Long;Ha, Do Thi;Lee, Myoung-Sook;Kim, Bo-Yeon;Luong, Hoang Van;Ahn, Jong-Seog;Bae, Ki-Hwan
    • Natural Product Sciences
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    • v.16 no.2
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    • pp.88-92
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    • 2010
  • We verified that the apoptosis activities were examined by DNA fragmentation, flow cytometric analysis with annexin V staining, activation of caspase-3, and cytochrome c release. In the result, $\alpha$- and $\beta$-eudesmol induced DNA fragmentation in HL-60 cells at a concentration of $80\;{\mu}M$, respectively. Additionally, pro-apoptotic cells sorted by flow cytometry analysis were detected in HL-60 cells to 31.77 and 29.67% with $\acute{a}$- and $\beta$-eudesmol of $80\;{\mu}M$. Thus, both $\alpha$- and $\beta$-eudesmol exerted caspase-3 activation and cytochrome c release at $80\;{\mu}M$ in HL-60 cells. These results are firstly reported that the sesquiterpenes, $\alpha$- and $\beta$-eudesmol are apoptosis inducers through mitochondria-dependent caspase cascade in HL-60 cells.

CD5+/CD21-Chronic Lymphocytic Leukemia in a Cat

  • Choi, Sorin;Bae, Hyeona;Chun, Daseul;Kim, Jihu;Shin, Sun Woo;Cho, ARom;Jung, Dong-In;Yu, DoHyeon
    • Journal of Veterinary Clinics
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    • v.37 no.6
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    • pp.350-354
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    • 2020
  • Feline chronic lymphocytic leukemia (CLL) is a rare disease. Its diagnosis is not simple because of the absence of clinical signs and the presence of mature lymphocytosis. An 11-year-old female spayed Russian Blue cat was referred to the veterinary medical teaching hospital for lethargy, diarrhea, weight loss, and inappetence. Marked lymphocytic leukocytosis and a significantly increased number of small-to-intermediate-sized lymphocytes in the peripheral blood were found on hematological examination. The results of the feline leukemia virus and immunodeficiency virus test were negative. Further, mild splenomegaly was detected. Bone marrow aspirate analysis revealed mature lymphocytosis and a clonally rearranged T cell receptor gene with the polymerase chain reaction (PCR) for antigen receptor rearrangement assay. Flow cytometric immunophenotyping showed a homogeneous population of CD5+/CD21-T-cells in the peripheral blood and bone marrow. According to the results of the aforementioned examinations, CLL was diagnosed. Treatment was not initiated at the time of diagnosis because the clinical signs were mild and did not affect the quality of life. This report describes the clinical findings and use of advanced diagnostic tools such as molecular clonality analysis and immunophenotyping for the diagnosis of feline CLL.

Effects of Fucoidan, a Sulfur-Containing Polysaccharide, on Cytotoxicity and Apoptosis in HT-29 Human Colorectal Cancer Cells (함유황 다당체 Fucoidan의 인체 대장암세포(HT-29) 사멸과 Apoptosis에 미치는 영향)

  • Kim, Min Ji;Chung, Ha Sook
    • The Korean Journal of Food And Nutrition
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    • v.35 no.3
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    • pp.204-212
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    • 2022
  • The purpose of this study was to investigate the biological activity of fucoidan, a sulfur-containing polysaccharide, on cytotoxicity and apoptosis in the human HT-29 colorectal cancer cell line using cell viability, Flow cytometry, Western blot, and RT-PCR analyses. Fucoidan inhibited the proliferation of HT-29 cells by 39.6% at a concentration of 100 ㎍/mL for 72 h. The inhibition was dose-dependent and accompanied by apoptosis. Flow cytometric analysis showed that fucoidan increased early apoptosis and late apoptosis by 65.84% and 72.09% at concentrations of 25 and 100 ㎍/mL, respectively. Analysis of the mechanism of these events indicated that fucoidan-treated cells exhibited increases in the activation of caspase-3, caspase-8, and PARP in a dose-dependent manner. These results suggest that fucoidan may inhibit the growth of human colorectal cancer cells by various apoptosis-promoting effects, as well as by apoptosis itself.

Peripheral Blood Lymphocytosis without Bone Marrow Infiltration in a Dog with T-Zone Lymphoma

  • YeSeul Jeon;Hyeona Bae;DoHyeon Yu
    • Journal of Veterinary Clinics
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    • v.40 no.3
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    • pp.203-208
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    • 2023
  • A 13-year-old neutered male mixed-breed dog presented with generalized lymphadenopathy and erythematous cutaneous lesions in the ear pinnae. Fine-needle aspiration cytology of the lymph nodes revealed small to intermediate lymphocytes with a "hand mirror" configuration as the predominant cell type. Histopathological analysis of the lymph node showed an infiltrate of CD3-positive small lymphocytes compressing the follicles against the capsule owing to neoplastic cell expansion. Flow cytometric analysis revealed a homogeneous population of CD3+/CD4-/CD5+/CD8-/CD21+/CD34-/CD45- cells in both the peripheral blood and aspirated lymph nodes, which supports the diagnosis of T-zone lymphoma. Laboratory tests revealed lymphocytosis (14,144 cells/µL) in the peripheral blood. However, contrary to expectations, the bone marrow examination revealed no evidence of lymphocytic infiltration. T-zone lymphoma is an indolent lymphoma with a long survival period, and knowledge of its characteristics may affect disease staging and prognosis evaluation. Therefore, peripheral blood count as a sole screening tool for bone marrow metastasis should be used with caution.

Anti-proliferative Effects of Water Extract of Agaricus blazei Murill in Human Lung Cancer Cell Line A549 (A549 인체폐암세포의 증식에 미치는 신령버섯 추출물의 영향에 관한 연구)

  • Choi, Woo-Young;Park, Cheol;Lee, Jae-Yun;Kim, Gi-Young;Park, Yeong-Min;Jeong, Yong-Kee;Lee, Won-Ho;Choi, Yung-Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.33 no.8
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    • pp.1237-1245
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    • 2004
  • Agaricus blazei Murill is a medicinal mushroom native to Brazil. It used to be a source of antitumor and immunoactive compounds and considered a health food in many countries. In the present study, it was examined the effects of water extract of A. blazei (WEAB) on the growth of human lung carcinoma cell line A549 in order to investigate the anti-proliferative mechanism by WEAB. Treatment of A549 cells to WEAB resulted in the growth inhibition, morphological change and induction of apoptotic cell death in a dose-dependent manner as measured by MTT assay and flow cytometric analysis. Flow cytometric analysis revealed that WEAB caused G2/M phase arrest of the cell cycle, which was associated with a down-regulation of cyclin A in both transcriptional and translational levels. WEAB treatment induced a marked up-regulation of cyclin-dependent kinase (Cdk) inhibitor p21, however, the levels of Cdk2, Cdc2, Wee1, Cdc25C and p53 expression were remained unchanged in WEAB treated cells. In addition, WEAB treatment inhibited the levels of cyclooxygenase (COX)-2 mRNA and protein without alteration of COX-l expression. Taken together, these findings suggest that WEAB may be a potential chemotherapeutic agent for the control of human lung carcinorma cells and further studies will be needed to identify the active compounds that confer the anti-cancer activity of WEAB. Once such compounds are identified, the mechanisms by which they exert their effects can begin to be characterized.

Angioimmunoblastic T-Cell Lymphoma with Polyclonal Proliferation of Plasma Cells: A Cautionary Note for Flow Cytometry Interpretations (유세포 분석의 주의사항: 혈관면역모세포성 T세포 림프종에서 관찰된 다클론성 형질세포)

  • Shin, Woo Yong;Bang, Hae In;Kim, Jung-Ah;Kim, Jieun;Park, Rojin
    • Korean Journal of Clinical Laboratory Science
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    • v.54 no.1
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    • pp.68-72
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    • 2022
  • Angioimmunoblastic T-cell lymphoma (AITL) is a lymphoproliferative disorder of mature T follicular helper cells. Atypical lymphoid cells were observed in the bone marrow of an 80-year-old woman, and the flow cytometric determined immunophenotypes of B-cells were unusual, that is, CD19+, CD20-, and CD22- with lambda light chain restriction. Initially, we suspected BM involvement of B-cell lymphoma based on the presence of abnormal B-cells. However, the patient was diagnosed with AITL involving BM. A re-analysis of flow cytometric immunophenotyping revealed a minor, aberrant T-cell population, and the lambda light chain restriction observed by surface staining was considered non-specific binding. This case demonstrates B-cells in patients with EBV-positive T-cell lymphoma may exhibit immunophenotypes resembling those of plasma cells, and that proliferation of abnormal B-cells or plasma cells could also potentially mask underlying T-cell lymphoma. A more integrated approach is required for accurate diagnosis.

Generation and Characterization of a Monoclonal Antibody with Specificity for Mycoplasma arginini

  • Son, Yeon-Sung;Hong, Hyo-Jeong
    • Journal of Microbiology
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    • v.45 no.6
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    • pp.547-552
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    • 2007
  • Previously, we generated monoclonal antibodies (MAbs) that bound to the surface of human embryonic stem cells (hESCs) in an attempt to discover new hESC-specific surface markers. In this study, MAb 47-235 (IgG1, ${\kappa}$) was selected for further characterization. The MAb bound to the surface of undifferentiated hESCs but did not bind to mouse ESCs or mouse embryonic fibroblast cells in flow cytometric analysis. The antibody immunoprecipitated a 47 kDa protein from the lysates of cell surface-biotinylated hESCs. Identification of the protein by quadrupole time of flight tandem mass spectrometry revealed that 47-235 binds to Ag 243-5 protein of Mycoplasma arginini. BM-Cyclin treatment of the hESCs that reacted with 47-235 resulted in loss of mycoplasma DNA and the reactivity to 47-235. Nevertheless, the hESCs that were reactive to 47-235 maintained self-renewal and pluripotency and thus could be differentiated into three embryonic germ layers.

Effects of Socheongryong-Tang on Inflammatory Cells in Asthmatic Mice (소청룡탕이 천식이 유발된 생쥐의 면역세포에 미치는 영향)

  • Kim, Hyung-Woo
    • Journal of Society of Preventive Korean Medicine
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    • v.10 no.2
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    • pp.121-129
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    • 2006
  • BACKGROUND : Socheongryong-Tang(小靑龍湯, SCRT), a herbal medicine, has been widely used for the control of respiratory disease in Eastern countries. Recent years SCRT was known as anti-allergic agent. However, its therapeutic mechanisms, especially an inhibitory effect on inflammatory cell infiltration and airway remodeling in allergic asthma are unclear. METHODS : We investigated the effects of SCRT on levels of IL-4, and IFN-g using flow cytometric analysis and infiltration of inflammatory cells in bronchoalveolar lavage fluid(BALF) using cytospin methods in allergen-induced asthma. RESULTS : SCRT decreased levels of IL-4, a Th2-driven cytokine. But INF-g, a Th1-driven cytokine, was not changed. With SCRT treatment, infiltration of inflammatory cells showed reduced tendency. CONCLUSION : In conclusion, we demonstrated that regulation of Th1/Th2 imbalance may be one of mechanism contributed to reduction of inflammatory cell infiltration.

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Craniospinal Metastasis from a Metastasizing Mixed Tumor of Salivary Gland : Unusual Presentation

  • Ye, Hyun-Hee;Cho, Chang-Won;Jeon, Mi-Young;Kim, Dae-Jo
    • Journal of Korean Neurosurgical Society
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    • v.41 no.3
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    • pp.186-189
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    • 2007
  • Metastasizing mixed tumors [MMT] of salivary glands are inexplicably metastasize maintaining benign histology. There is no pathologic and flow cytometric analysis criteria to predict the metastasis. MMT is known to metastasize by local implantation, vascular and lymphatic embolization after multiple surgery to local recurrences of primary tumor. However, multiple metastasis including cranium and spine occurred even without surgery to the primary tumor in this case. No pathological evidence of malignancy could be found in both primary and metastatic tumor. MMT is considered as an low grade malignancy based on clinical behavior rather than histologic evidence, such as low mortality rate, long delay of metastasis after primary lesion. Cranial metastasis is also extremely rare and only two cases have been reported. We report this unusual case with a literature review.

Bio-inert Surface of Pluronic-immobilized Flask for Preservation of Hematopoietic Stem Cells

  • Higuchi, Akon;Aoki, Nobuo;Gomei, Yumiko;Matsuoka, Yuki
    • Proceedings of the Polymer Society of Korea Conference
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    • 2006.10a
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    • pp.267-267
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    • 2006
  • Human umbilical cord blood was stored at $4^{\circ}C$ in the Pluronic-immobilized flask as well as commercially available bio-inert flasks, and flow cytometric analysis of surface markers was performed on hematopoietic stem cells after cultivation. The number of cells expressing $CD34^{+}$ in umbilical cord blood on the Pluronic-immobilized flask was extremely higher than those obtained using other flasks. It is concluded that the flexible and hydrophilic segments of Pluronic conjugated on the flask surface are the reason for the effective preservation of hematopoietic stem cells in the Pluronic-immobilized flask.

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