• Title/Summary/Keyword: Flow cytometric analysis

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Diagnostic Value of Flow Cytometric DNA Analysis in the Evaluation of Effusions (체강삼출액의 진단에 있어서 유세포분석에 의한 DNA 함량 측정의 유용성)

  • Lee, Ji-Shin;Juhang, Sang-Woo
    • The Korean Journal of Cytopathology
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    • v.8 no.1
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    • pp.20-26
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    • 1997
  • The specificity of cytologic examination in effusions is high but the sensitivity is low. Therefore, various ancillary methods for the detection of malignant cells in effusions have been proposed. The presence of an aneuploid cell population is generally considered diagnostic of malignancy. The purpose of this study is to determine whether the routine use of flow cytometry adds to standard cytologic evaluation in effusions. We did flow cytometric DNA analysis in 76 effusions(28 malignant and 48 benign fluids). All the 48 benign effusions were diploid. There were 12(42.9%) aneuploid and 16(67.1%) diploid malignant effusions. Based on these results flow cytometric DNA analysis had a sensitivity of 42.9% and a specificity of 100%. These results suggest that flow cytometric DNA analysis may be a useful adjunct to conventional cytology, but its principal limitation is us relatively low sensitivity.

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Flow Cytometric Analysis of Human Lysozyme Production in Recombinant Saccharomyces cerevisiae

  • Peterson Marvin S.;Kim Myoung-Dong;Han Ki-Cheol;Kim Ji-Hyun;Seo Jin-Ho
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.7 no.1
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    • pp.52-55
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    • 2002
  • Flow cytometric techniques were used to investigate cell size, protein content and cell cycle behavior of recombinant Saccharomyces cerevisiae strains producing human lysozyme (HLZ). Two different signal sequences, the native yeast $MF\alpha1$ signal sequence and the rat $\alpha-amylase$ signal sequence, were used for secretion of HLZ. The strain containing the rat $\alpha-amylase$ signal sequence showed a higher level of internal lysozyme and lower specific growth rates. Flow cytometric analysis of the total protein content and cell size showed the strain harboring the native yeast signal sequence had a higher total protein content than the strain containing the rat $\alpha-amylase$ signal sequence. Cell cycle analysis indicated that the two lysozyme producing recombinant strains had an increased number of cells in the $G_2+M$ phase of the yeast cell cycle compared with the host strain SEY2102.

Flow Cytometric Analysis of the Effect of Silkworm Hemolymph on the Baculovirus-Induced Insect Cell Apoptosis

  • Rhee, Won-Jong;Park, Tai-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.11 no.5
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    • pp.853-857
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    • 2001
  • The effect of silkworm hemolymph on the inhibition of baculovirus-induced insect cell apoptosis was quantitatively investigated using a flow cytometric analysis. Spodoptera frugiperda (Sf9) cell and Autographa californica nuclear polyhedrous virus (AcNPV) were used as a model for insect cell and baculovirus in this study, respectively. Compared with a mammalian cell cycle, the fraction of G1 cells was relatively small in the Sf9 cell cycle. Silkworm hemolymph did not affect the Sf9 cell-cycle distribution before the baculovirus infection. However, the fraction of cells which are not in the sub-G1 phase remained at a high level for 3 days after the infection in the medium without silkworm hemolymph, while it remained at a high level for 7 days after the infection in the medium supplemented with silkworm hemolymph. The fractions of apoptotic cells in the sub-G1 phase were $4.7\%$, and 4 days after infection, $22.7\%$, in the media with and without silkworm hemolymph, respectively.

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A Rapid Assessing Method of Drug Susceptibility Using Flow Cytometry for Mycobacterium tuberculosis Isolates Resistant to Isoniazid, Rifampin, and Ethambutol

  • Lee, Sun-Kyoung;Baek, Seung-Hun;Hong, Min-Sun;Lee, Jong-Seok;Cho, Eun-Jin;Lee, Ji-Im;Cho, Sang-Nae;Eum, Seok-Yong
    • Tuberculosis and Respiratory Diseases
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    • v.85 no.3
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    • pp.264-272
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    • 2022
  • Background: The current conventional drug susceptibility test (DST) for Mycobacterium tuberculosis (Mtb) takes several weeks of incubation to obtain results. As a rapid method, molecular DST requires only a few days to get the results but does not fully cover the phenotypic resistance. A new rapid method based on the ability of viable Mtb bacilli to hydrolyze fluorescein diacetate to free fluorescein with detection of fluorescent mycobacteria by flow cytometric analysis, was recently developed. Methods: To evaluate this cytometric method, we tested 39 clinical isolates which were susceptible or resistant to isoniazid (INH) or rifampin (RIF), or ethambutol (EMB) by phenotypic or molecular DST methods and compared the results. Results: The susceptibility was determined by measuring the viability rate of Mtb and all the isolates which were tested with INH, RIF, and EMB showed susceptibility results concordant with those by the phenotypic solid and liquid media methods. The isolates having no mutations in the molecular DST but resistance in the conventional phenotypic DST were also resistant in this cytometric method. These results suggest that the flow cytometric DST method is faster than conventional agar phenotypic DST and may complement the results of molecular DST. Conclusion: In conclusion, the cytometric method could provide quick and more accurate information that would help clinicians to choose more effective drugs.

Multiparameter Flow Cytometry: Advances in High Resolution Analysis

  • O'Donnell, Erika A.;Ernst, David N.;Hingorani, Ravi
    • IMMUNE NETWORK
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    • v.13 no.2
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    • pp.43-54
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    • 2013
  • Over the past 40 years, flow cytometry has emerged as a leading, application-rich technology that supports high-resolution characterization of individual cells which function in complex cellular networks such as the immune system. This brief overview highlights advances in multiparameter flow cytometric technologies and reagent applications for characterization and functional analysis of cells modulating the immune network. These advances significantly support highthroughput and high-content analyses and enable an integrated understanding of the cellular and molecular interactions that underlie complex biological systems.

Flow Cytometric DNA Analysis of Thyroid Papillary Carcinoma (갑상선 유두상암의 핵산분석)

  • Chu Hyung-Ro;Jung Do-Kwang;Woo Jeong-Su;Choi Jong-Ouck;Kim In-Sun
    • Korean Journal of Head & Neck Oncology
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    • v.10 no.2
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    • pp.102-105
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    • 1994
  • The indolent course of most thyroid papillary carcinomas, even the presence of regional lymph node metastasis, make them unique among human malignant head and neck cancers. Age, sex, extracapsular invasion and anaplastic change are known to be correlated with prognosis. The purpose of this study is to clarify the significance of DNA content analysis as a prognostic factor. Twenty five thyroid papillary carcinomas were possible to be examined by flow cytometric analysis using fresh surgical specimens and three nodular hyperplasias and seven follicular adenomas were included as control group. The results were as follows: l) All of twenty five thyroid papillary carcinomas showed diploidy. 2) S-phase fraction was $1.94{\pm}2.77%$ in normal control group and $2.60{\pm}2.66%$ in papillary carcinoma group. The proliferation index was $8.44{\pm}3.89%$ in normal control and $7.70{\pm}3.63%$ in papillary carcinoma group with even low value. 3) Age, sex, extracapsular spread and lymph node metastasis showed no significant difference. In conclusion, low proliferative activity of thyroid papillary carcinomas are thought to be related with good prognosis.

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Comparison of polymerase chain reaction for antigen receptor gene rearrangement and flow cytometric analysis for the diagnosis of canine lymphoma

  • Song, Ru-Hui;Yu, Do-Hyeon;Kim, Jun-Hwan;Lee, Hyun-Seok;Lee, Da-Mi;Park, Chul;Yu, Il-Jung;Park, Jin-Ho
    • Korean Journal of Veterinary Service
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    • v.34 no.3
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    • pp.265-271
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    • 2011
  • Lymphoma is the most common hematopoietic malignancy in dogs. Diagnosis of lymphoma is classically performed by morphological assessment and immunohistochemistry. But some cases in the early stage are difficult to distinguish and need more objective and accurate methods. So, Polymerase chain reaction (PCR) for antigen receptor rearrangements (PARR) and flow cytometric immunophenotype of lymphoma have been developed continuously. In this study, we performed these two methods to classify lymphoma type in 3 cases. According to PARR analysis, B cell origin lymphoma was diagnosed in two of three cases by testing PBMC and lymph node. All fine needle aspiration (FNA) samples of lymph nodes had high expression of CD21 on >88% of total cell population and PBMC samples also showed high expression of CD21 on >30% of total lymphocytes in those two cases, while the expression of CD3, CD4 and CD8 was absent. These results suggest that concurrent use of PARR and flow cytometric immunophenotype is more effective and valuable tool for the diagnosis and monitoring of canine lymphoma patients.

AN EXPERIMENTAL STUDY ON MISTLETOE EXTRACT-INDUCED APOPTOSIS IN ORAL SQUAMOUS CELL CARCINOMA (구강편평세포암종에서 미슬토 추출물의 Apoptosis 유도 효과에 대한 실험적 연구)

  • Heo, Gyun-Haeng;Lee, Jae-Hoon;Kim, Chul-Hwan
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.1
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    • pp.13-23
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    • 2005
  • This study was performed to investigate mistletoe extract-induced apoptosis in oral squamous cell carcinoma. In vivo study, HN22 cells were xenografted in nude mice. After tumor was experimentally induced, mistletoe extract was directly injected on the tumor mass. The specimens were evaluated using light and transmission electron microscopes. In vitro study, HN22 cells were cultured and exposed to mistletoe extract. The cells were evaluated using transmissin electron microscope. To evaluate apoptotic cells, flow cytometric analysis was done. The results were obtained as follows: 1. Light microscopic view of tumor mass showed necrosis at 2-4 weeks. 2. Transmission electron micrographs of tumor mass showed apoptosis and necrosis. 3. In TEM view of cell lines, necrosis and apoptosis were shown with mistletoe extract at $300{\mu}g/ml$, apoptosis was shown with mistletoe extract at $100{\mu}g/ml$. 4. In flow cytometric analysis, early and late apoptosis was shown when using caspase-3Ab and annexin-V, but no significant change was noted when using mebstain and Apo2.7 Ab. In this study, mistletoe extract induced necrosis and apoptosis in the tumor mass was induced by HN22 cells, early and late apoptosis in vitro study. Mistletoe extract was likely to induce cell death in oral squamous cell carcinoma through apoptosis.

Prognostic significance of minimal residual disease detected by a simplified flow cytometric assay during remission induction chemotherapy in children with acute lymphoblastic leukemia

  • Koh, Kyung-Nam;Park, Mee-Rim;Kim, Bo-Eun;Im, Ho-Joon;Park, Chan-Jeoung;Jang, Seong-Soo;Chi, Hyun-Sook;Seo, Jong-Jin
    • Clinical and Experimental Pediatrics
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    • v.53 no.11
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    • pp.957-964
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    • 2010
  • Purpose: Our study attempted to determine the prognostic significance of minimal residual disease (MRD) detected by a simplified flow cytometric assay during induction chemotherapy in children with B-cell acute lymphoblastic leukemia (B-ALL). Methods: A total of 98 patients were newly diagnosed with precursor B-ALL from June 2004 to December 2008 at the Asan Medical Center (Seoul, Korea). Of those, 37 were eligible for flow cytometric MRD study analysis on day 14 of their induction treatment. The flow cytometric MRD assay was based on the expression intensity of CD19/CD10/CD34 or aberrant expression of myeloid antigens by bone marrow nucleated cells. Results: Thirty-five patients (94.6%) had CD19-positive leukemic cells that also expressed CD10 and/or CD34, and 18 (48.6%) had leukemic cells with aberrant expression of myeloid antigens. Seven patients with ${\geq}1%$ leukemic cells on day 14 had a significantly lower relapse-free survival (RFS) compared to the 30 patients with lower levels (42.9 % [18.7%] vs. 92.0% [5.4%], $P$=0.004). Stratification into 3 MRD groups (${\geq}1%$, 0.1-1%, and <0.1%) also showed a statistically significant difference in RFS (42.9% [18.7%] vs. 86.9% [8.7%] vs. 100%, $P$=0.013). However, the MRD status had no significant influence on overall survival. Multivariate analysis demonstrated that the MRD level on day 14 was an independent prognostic factor with borderline significance. Conclusion: An MRD assay using simplified flow cytometry during induction chemotherapy may help to identify patients with B-ALL who have an excellent outcome and patients who are at higher risk for relapse.

The Effects of Orostachys japonicus on HepG2 Cell Proliferation and Oncogene Expression (와송(瓦松)이 HepG2 cell의 세포분열 및 관련유전자 발현에 미치는 영향)

  • Moon, Young-Hun;Kim, Young-Chul;Lee, Jang-Hoon;Woo, Hong-Jung
    • The Journal of Internal Korean Medicine
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    • v.26 no.1
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    • pp.48-59
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    • 2005
  • Objectives : The aim of the study was to evaluate the effect of WS on HepG2 cell cycle and expression of related genes. Methods : The MTT assay, Cell counting analysis, $[^3H]-Thymidine$ Incorporation Assay, Flow cytometric analysis, Quantitative RT-PCR were studied. Results : WS inhibited HepG2 cell proliferation in low concentration$(1-10\;{\mu]g/ml)$ which did not cause direct cytotoxicity, with dose-dependant manner. WS in-hibited DNA synthesis as well. Flow cytometric analysis on the HepG2 cell showed G2/M phase arrest. Conclusion : These results suggest that WS inhibits HepG2 cell proliferation not by the gene regulation but by G2/M phase arrest in the cell cycle. Thus further studies on the effect of WS in G2/M phase regulation are thought to be needed.

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