• Title/Summary/Keyword: Nitroblue tetrazolium

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Effects of stress induced by changes of water temperature on the non-specific defense mechanism in cultured olive flounder, Paralichthys olivaceus (수온 변동 자극이 양식 넙치, Paralichthys olivaceus의 비특이적 생체 방어에 미치는 영향)

  • Lee, Deok-Chan;Kim, Do-Hyung;Kim, Su-Mi;Kang, Myong-Seok;Hong, Mi-Ju;Kim, Hyun-Jeong;Park, Soo-Il
    • Journal of fish pathology
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    • v.15 no.2
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    • pp.65-75
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    • 2002
  • This study was performed to know the effects of stress induced by the daily fluctuation of water temperture from 18$^{\circ}C$ to 25$^{\circ}C$ up and down for 30 days on the defence mechanism of olive flounder. Puralichthys olivaceus. To make clear the temperature stress on the defense mechanism of the tested fish. several factors of immune response such as counting of leucocyte appearance in peripheral blood, phagocytic activity in whole blood cells, nitroblue tetrazolium(NBT) reduction, chemiluminescence(CL) response, and lysozyme activity were investigated at 28 days after giving the change of water temperature. The fish was controlled under the none feeding condition during experimental period. Mortality of the tested fish was rapidly increased up to 22% within the first one week of the experimental period without any additional stress factors. The number of neutrophil of peripheral blood in the tested group was significantly higher than the control group at the 2nd week. but the number of lymphocyte was significantly lower than the control group at the 1st and 3rd day of the experimental period. respectively. In the NBT reduction test, the activity of macrophage in the control group fish was the highest on the 7th day while that in the tested group was on the 3rd day. Also. the phagocytosis of tested group against formalin killed cells was retarded compared with the control. CL response of the tested group was significantly lower from 2nd to 5lh day of the experimental period than the control. 'The lysozyme activity of tested group was remained higher during the experimental period than the control. Even though the tested fish showed different results in some non-specific factors of immune respceses between tested and control group fish, olive flounder seems highly adaptable in repealed water temperature change in condition after one week under the given temperature fluctuation range.

Cytotoxicity in HL-60 cells and human lymphocytes and effect of leukemia cell differentiation induced by Saussureae Radix extract (정상사람림프구와 HL-60 cell에서 목향의 세포독성과 백혈병세포 분화효과에 관한 연구)

  • Lee, Young-Joon;Kang, Su-Jin;Ku, Sae-Kwang
    • The Korea Journal of Herbology
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    • v.26 no.2
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    • pp.31-37
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    • 2011
  • Objectives : This study was focused to investigate the toxicity of Saussurea lappa (SL) extracts in HL-60 cells and human lymphocytes. We also examined the differentiation effect of SL against leukemia cells. Methods : For examining the toxicity of SL, cytokinesis-block micronucleus (CBMN) assay and single cell gel eletrophoresis (SCGE) assay were used in present study. The cell differentiation effect of SL was evaluated by nitroblue tetrazolium (NBT) reduction assay. Results : The inhibition of cell growth in HL-60 cells was observed in a dose-dependant manner after SL treatment for 24 h. According to SCGE assay, HL-60 cells treated with SL increased DNA damage at $10{\mu}g/m{\ell}$, while DNA damage was induced by 0.1, 1, $10{\mu}g/m{\ell}$ concentration of SL in human lymphocytes. Our results indicated that SL have no genotoxic effect in HL-60 cells and human lymphocytes. Additionally, the differentiation effect was induced in $1{\mu}g/m{\ell}$ SL-treated HL-60 cells. Conclusions : From above results it is suggested that SL could be beneficial for the preparation of the useful agent for treating leukemia.

Effects of feeding of dried Euglena rubra on Nile tilapia Oreochromis niloticus (빨간유글레나, Euglena rubra 건조분말의 사료첨가에 따른 나일틸라피아, Oreochromis niloticus에서의 영향)

  • Lee, Jae-Geun;Lee, Han-Na;Kim, Young-Dae;Choi, Sang-Hoon;Park, Kwan-Ha
    • Journal of fish pathology
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    • v.24 no.3
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    • pp.307-313
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    • 2011
  • To examine the effects of Euglena rubra (E. rubra) feeding on Nile tiapia, Oreochromis niloticus were fed diets containing dried powder of E. rubra at 0.5 or 2% for 4 weeks. Growth, selected hematological and non-specific immune parameters were assessed at 2 and 4 weeks. There were no significant changes in body weigh gain and erythrocyte levels. However a significant and diet E. rubra level-related decrease in leukocyte level was noted. Significant increases were observed in respiratory burst activity (nitroblue tetrazolium reduction) and lysozyme activities following E. rubra feeding. These results indicate that E. rubra could be beneficial to fish, but excess feeding could toxic.

Induction of Differentiation on the Human Histocytic Lymphoma Cell Line U-937 by Costunolide (Costunolide의 백혈병 세포주 U-937에 대한 분화 유도 작용)

  • Kim, Joo-Il;Lee, Sung-Ho;Park, Jae-Hoon;Park, Hee-Joun;Lee, Kyung-Tae
    • Korean Journal of Pharmacognosy
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    • v.30 no.1
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    • pp.7-11
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    • 1999
  • The present work was carried out to examine the effect of costunolide on the growth of several cells and characteristics of U-937 human leukemia-derived cell line. Costunolide produced a potent antitumor activity in vitro dependent on concentration against several tumor cells such as P-388, L-1210 leukemia and SNU-5 stomach cancer cells. However, it showed less cytotoxicity on normal cells such as Maccaccus rheus monkey kidney cells (MA-104) up to 200 ${\mu}M$ concentration. An effect of cell differentiation by costunolide was assessed by its ability to reduce nitroblue tetrazolium (NBT), and to induce phagocytosis of latex particles. In order to establish whether costunolide induces U-937 cells to differentiate toward macrophage or granulocyte, esterase activities was measured. Based on these results, we found that costunolide having cytotoxicity on U-937 human leukemia cells was explained through differentiation inducing activity.

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Endotoxins of Enteric Pathogens Modulate the Functions of Human Neutrophils and Lymphocytes

  • Islam, Laila N.;Nabi, A.H.M. Nurun
    • BMB Reports
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    • v.36 no.6
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    • pp.565-571
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    • 2003
  • The locomotor responses of human peripheral blood neutrophils and lymphocytes were measured by the change from spherical to polarized shapes in the presence of endotoxins (lipopolysaccharide, LPS) of enteric pathogens: S. dysenteriae type 1, V. cholerae Inaba 569B, S. typhimurium, and K. pneumoniae. We reported earlier that these endotoxins are chemotactic factors for the neutrophils since they stimulated cell polarization within a few minutes of incubation. Endotoxins had an inhibitory effect upon neutrophil phagocytosis of opsonized yeast and the cells engulfed fewer yeasts. Interestingly, endotoxins increased neutrophil adhesion to clean glass surfaces, but stimulated the cells to exhibit increased random locomotion (chemokinesis) through cellulose nitrate filters and show an enhanced ability to reduce nitroblue tetrazolium (NBT) dye. Unlike neutrophils, lymphocytes direct from blood do not show polarized morphology towards chemotactic factors but the cells acquire locomotor capacity during 24-72 h culture with mitogens such as phytohemagglutinin (PHA), phorbol myristate acetate or concanavalin A. Stimulation of blood lymphocytes with endotoxins did not induce cell polarization in short-term but long-term culture resulted in an increase in the proportion of polarized cells that acquired locomotor morphologies. The majority of these cells were identified as esterase negative B-lymphocytes that migrated through filters. Despite the optimum time of incubation for each of these cell types being different, we found that lymphocytes respond to much lower concentrations of endotoxins than the neutrophils. These findings suggest that endotoxins of enteric pathogens modulate the functions of human blood neutrophils and lymphocytes.

PBK/TOPK Expression During TPA-Induced HL-60 Leukemic Cell Differentiation

  • Liu, Yu-Hong;Gao, Xue-Mei;Ge, Fan-Mei;Wang, Zhe;Wang, Wen-Qing;Li, Xiao-Yong
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.5
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    • pp.2145-2148
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    • 2012
  • Objective: This study concerns expression of PBK/TOPK during differentiation of HL-60 leukemic cells induced by tetradecanoyl phorbol acetate (TPA). Methods: Wright-Giemsa staining was performed to observe morphological changes in the HL-60 cells, and flow cytometry was used to assess the cell cycle and CD11b, CD14, CD13, and CD33 expression. PBK/TOPK levels were determined by Western blot analysis. Results: After treating HL60 cells with $5.1{\times}10^{-9}$ mmol/L of TPA for three days, the number of nitroblue-tetrazolium-positive cells and CD11b, CD13, and CD14 expression increased, whereas the PBK/TOPK levels decreased. Conclusions: TPA can inhibit proliferation and induce differentiation of HL60 cells of the granulocytic or monocytic lineage. PBK/TOPK expression was downregulated during this process, whereas the Pho-PBK/TOPK expression was increased.

Induction of Quinone Reductase and Glutathione S-Transferase in Murine Hepatoma Cells by Flavonoid Glycosides

  • Kim, Jung-Hyun;Lee, Jeong-Soon;Kim, Young-Chan;Chung, Shin-Kyo;Kwon, Chong-Suk;Kim, Young-Kyoon;Kim, Jong-Sang
    • Preventive Nutrition and Food Science
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    • v.8 no.4
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    • pp.365-371
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    • 2003
  • The potential of seven flavonoid glycosides to induce quinone reductase (QR), an anticarcinogenic marker enzyme, in murine hepatoma cells (hepalc1c7) and its mutant cells (BPRc1) was evaluated. Among test compounds, kaempferol-3-O-glucoside, luteolin-6-c-glucoside, and quercetin-3-O-glucoside (Q-3-G) induced QR in hepalc1c7 cells in a dose-dependent manner. However, in BPRc1 cells lacking arylhydrocarbon receptor nuclear translocator (ARNT), only Q-3-G caused a significant induction of quinone reductase at the concentration range of 0.5 to 8 ug/mL, suggesting that it is a monofunctional inducer. Q-3-G induced not only phase 2 enzymes, including QR and glutathione-S-transferase, but also nitroblue tetrazolium reduction activity in HL-60 cells, a biochemical marker for cell differentiation promoting agents. In conclusion, Q-3-G merits further study to evaluate its cancer chemopreventive potential.

Application of alkaline phosphatase staining of cytology specimen for differential diagnosis of canine osteosarcoma (세포 검사시료에서 alkaline phosphatase 염색법을 활용한 개 골육종의 감별 진단)

  • Park, Byoung-Yong;Park, Chul;Cho, Ho-Seong
    • Korean Journal of Veterinary Service
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    • v.34 no.4
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    • pp.397-401
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    • 2011
  • Aspiration of lytic bone lesions is an excellent diagnostic test in the initial evaluation of primary bone tumor. However, cytologically, it can be difficult to differentiate osteosarcoma (OSA) from other bone neoplasms, including fibrosarcoma, chondrosarcoma, synovial cell sarcoma, malignant fibrous histiocytoma and malignant peripheral nerve sheath tumor. The purpose of this study is to introduce alkaline phosphatase (ALP) staining to differentiate OSA from other mesenchymal tumors. Tumors actively producing bone are specifically positive for ALP staining. Unstained, cytologic specimens were incubated for 10 minutes with nitroblue tetrazolium chloride/5-bromo-4-chloro-3-indolyl phosphate toluidine salt-phosphatase substrate. Among 20 cases of cytology specimen, 14 were positive for ALP staining and histopathology, 6 were negative for ALP staining and histopathology. ALP staining was 100% sensitive and specificity for the diagnosis of OSA. Aspirate cytology with ALP staining was a simple, fast, safe and accurate diagnostic test for the evaluation of suspected OSA lesions in dogs.

Induction of Differentiation of the Human Histocytic Lymphoma Cell Line U-937 by Hypericin

  • Kim, Joo-Il;Park, Jae-Hoon;Park, Hee-Juhn;Choi, Seung-Ki;Lee, Kyung-Tae
    • Archives of Pharmacal Research
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    • v.21 no.1
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    • pp.41-45
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    • 1998
  • Hypericin, a photosensitizing plant pigment, was found to be a potent inducer of differentiation of human myeloid leukemia U-937 cells. At a concentration of $0.2{\mu}M$, hypericin exhibited 50% growth inhibition. An effect on cell differentiation by hypericin was assessed by its ability to induce phagocytosis of latex particles, and to reduce nitroblue tetrazolium (NBT). Approximately 51% of $0.2{\mu}M$ hypericin-treated cells were stained with NBT and 63% showed phagocytic activity. In order to establish whether hypericin induces differentiation of U-937 cells to macrophage or granulocyte, esterase activities and cell sizes were measured. When U-937 cells were treated with $0.2{\mu}M$ and $0.15{\mu}M$ of hypericin, the .alpha.-naphthyl acetate esterase activity was increased by 38.4% and 48.1%, respectively, but naphthol AS-D chloroacetate esterase activity was not influenced. The size of hypericin-treated cells in terms of cell mass was larger than that observed in untreated cells as determined by flow cytometry. Protein kinase C (PKC) inhibitor, NA-382, decreased the NBT reducing activity of hypericin, whereas a cAMP-dependent protein kinase A (PKA) inhibitor, H-89, did not show any influence on the differentiations. These results indicate that hypericin triggers differentiation toward monocyte/macrophage lineage by PKC stimulation.

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Maltol, an Antioxidant Component of Korean Red Ginseng, Shows Little Prooxidant Activity

  • Suh, Dae-Yeon;Han, Yong-Nam;Han, Byung-Hoon
    • Archives of Pharmacal Research
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    • v.19 no.2
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    • pp.112-115
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    • 1996
  • Some antioxidant phenolic compounds exhibit prooxidant activity mainly due to their abilities to reduce $Fe^{3+}\; to\; Fe^{2+}.$ Reducing ability and prooxidant activity of maltol, an antioxidant component of Korean red ginseng, were compared with those of pyrogallol. Maltol at 2 mM did not appreciably reduce$ Fe^{3+}\; to\; Fe^{2+}$ and also failed to reduce nitroblue tetrazolium. Stimulation of hydroxyl radical mediated-deoxyribose degradation by pyrogallol was maximal at 60 .mu.M. Maltol stimulated the deoxyribose degradation to a much less extent, and a similar stimulatory effect was observed at a concentration of more than 100-fold higher than that of pyrogallol. The stimulatory effect of maltol reached a plateau over 1 mM, suggesting the removal of hydroxyl radicals by excess maltol. In bleomycin-$Fe^{3+}$-DNA assay, maltol at 2 mM produced a 2.5-fold increase of the iron-bleomycin-dependent DNA degradation over the basal value, whereas pyrogallol at 10 .mu.M accelerated DNA degradation by ca. 10-fold. Furthermore, maltol inhibited $Fe^{2+}$-stimulated DNA degradation by bleomycin. These results strongly suggested that maltol is an antioxidant with little prooxidant activity.

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