• Title/Summary/Keyword: NBT reduction

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Effect of Rhizoma Zingiberis Siccatum Extract on the Induction of HL-60 Human Leukemia Cell Differentiation (HL-60 세포에 대한 건강의 분화유도 효과)

  • Oh, Yu-Mi;Ju, Seong-Min;Kim, Young-Mi;Yun, Yong-Gab;Jeon, Byung-Hun;Kim, Won-Sin
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.2
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    • pp.345-351
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    • 2006
  • We have examined the induction of HL-60 cell differentiation by treatment of the water extract of Rhizoma Zingiberis Siccatum, which is a Korean traditional herbal medicine. It was observed that HL-60 cell proliferation was dose-dependently inhibited by treatment with various dose of Rhizoma Zingiberis Siccatum extract. The level of inhibition was identical to those of ATRA-treated cells. Rhizoma Zingiberis Siccatum extract treatment caused a significant change in NBT reduction (47%) and enhanced ATRA-induced NBT reduction. Treatment of Rhizoma Zingiberis Siccatum to HL-60 cells increased only CD11 b expression in the cells, and also increased markedly G0/G1 stage arrest. This can suggest that Rhizoma Zingiberis Siccatum induced the differentiation of HL-60 cell and enhanced ATRA-induced differentiation predominantly along the granulocytic lineage. The results present here show that Rhizoma Zingiberis Siccatum extract contains the potential effect of differentiation induction or acts as a agent enhancing the induction of differentiation.

Effect of 2-Chloromethyl-1-Dihydroxyphosphinylpyrrolidine(2C-1DPP) on Differentiation Induction of Human Leukemia HL-60 Cells (2-Chloromethyl-1-Dihydroxyphosphinylpyrrolidine (2C-1DPP)에 의한 백혈병 세포주 HL-60의 분화유도 효과)

  • Kim, Youg-Mi;Ju, Seong-Min;Park, Jun-Ho;Oh, Jung-Mi;Lee, Chae-Ho;Kim, Eun-Cheol;Jeon, Byung-Hun;Kim, Won-Sin;Kim, Won-Sin
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.4
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    • pp.940-945
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    • 2007
  • We have examined the induction of HL-60 cell differentiation by treatment of 2-chloromethyl-1-dihydroxyphosphinyl pyrrolidine(2C-1DPP), which is derivative of piperidine and pyrrolidine by ${\alpha}-phosphoramidoakylation$ reaction. It was observed that HL-60 cell proliferation was dose- and time-dependently inhibited by treatment with 2C-1DPP. 2C-1DPP treatment caused a significant change in NBT reduction and enhanced ATRA-induced NBT reduction. Treatment of 2C-1DPP to HL-60 cells increased only CD11b expression in the cells, and also increased markedly G0/G1 stage arrest of HL-60 cells. These results can suggest that 2C-1DPP induced the differentiation of HL-60 cells to granulocytes lineage and enhanced ATRA-induced differentiation. Moreover, DNA expression levels of p27 were up-regulated during 2C-1DPP-dependent HL-60 cell differentiation. Our results suggest that 2C-1DPP have potential as a therapeutic agent in human leukemia.

Generation of Superoxide Radical from Rat Brain Mitochondria and Mechanism of Its Toxic Action to Mitochondrial and Extra-mitochondrial Components (흰쥐 뇌 미토콘드리아에 의한 superoxide radical의 생성과 이 radical이 미토콘드리아 및 미토콘드리아 외 물질에 대한 독작용과 그 기전에 관한 연구)

  • Roh, Jae-Kyu;Pyo, Jang-Geun;Chung, Myung-Hee;Lim, Jung-Kyoo;Myung, Ho-Jin
    • The Korean Journal of Pharmacology
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    • v.21 no.1
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    • pp.12-26
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    • 1985
  • The generation of $O^{-}_{2}{\cdot}$ and its toxic effects were studied with rat brain mitochondria. The production of $O^{-}_{2}{\cdot}$ from mitochondria in the presence of succinate and antimycin was demonstrated by SOD-inhibitable reduction of NBT. Although succinate can support the $O^{-}_{2}{\cdot}$ formation, the highest rate needs antimycin indicating that blockade of electron flow in the respiratory chain augments the univalent reduction of molecular oxygen. Under this condition, $H_2O_2$ was also observed to be produced. But its formation appears to be derived from the dismutation of the primary product, $O^{-}_{2}{\cdot}$ since the rate of $H_2O_2$ production was markedly decreased by NBT and ferricytochrome c. The $O^{-}_{2}{\cdot}$ and $H_2O_2$ produced were able to cause toxic actions to mitochondrial and extra-mitochondrial components as shown by lipid peroxidation of mitochondrial membrane, and inactivation and lysis of isocitrate dehydrogenase and erythrocytes added to the medium, respectively. In all the toxic actions observed, $Fe^{++}$ was required. It appears that in the toxic actions $OH{\cdot}$ generated from the iron-catalyzed Haber-Weiss reaction acts as a mediator. This was supported by the finding that mitochondria in the presence of succinate and antimycin produced ethylene from methional, and $Fe^{++}$ added increased the ethylene production. The observed toxic actions of mitochondrial $O^{-}_{2}{\cdot}$ may provide evidence supporting a potential role of mitochondria as a source of oxygen radicals to cause tissue damage.

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Endotoxins of Enteric Pathogens Are Chemotactic Factors for Human Neutrophils

  • Islam, Laila N.;Nabi, A.H.M. Nurun;Ahmed, K. Mokim;Sultana, Novera
    • BMB Reports
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    • v.35 no.5
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    • pp.482-487
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    • 2002
  • Early activation of human peripheral blood polymorphonuclear neutrophils is characterized by their morphological changes from spherical to polarized shapes. The endotoxins from enteric pathogens (S. dysenteriae type 1, V. cholerae Inaba 569B, S. typhimurium, and K. pneumoniae) were assessed by their ability to induce morphological polarization of the neutrophils as measures of early activation. Phagocytic activity, adhesion, chemokinetic locomotion, and nitroblue tetrazolium (NBT) dye-reduction ability measured the later activation of the cells. Neutrophils showed distinct morphological polarization in suspension over a wide range of concentrations of these endotoxins when were compared with those that were induced by the standard chemotactic factor, N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP). It was discovered that all of the endotoxins induced locomotor responses in neutrophils in suspension that were dose- and time-dependent. The optimum concentration for the endotoxins of S. dysenteriae, V. cholerae, and K. pneumoniae was 1 mg/ml in which 71, 69, and 66% of the neutrophils were polarized. However, the S. typhimurium dose was 2 mg/ml in which 50% of the cells responded. Neutrophils that were stimulated with endotoxins also showed increased random locomotion (p<0.005) through cellulose nitrate filters, but an enhanced adhesion of the cells to glass surfaces (p<0.03). These are important functions of these cells to reach and phagocytose damaged cells, as well as invading microorganisms. Interestingly, the endotoxins had a highly-significant inhibitory effect upon the proportions of neutrophils phagocytosing opsonized yeast (p<0.01) with a small number of yeast that were engulfed by the cells (p<0.02). Further, endotoxin-treated cells showed an enhanced ability to reduce NBT dye (p<0.03). Therefore, we concluded that endotoxins of enteric pathogens are neutrophil chemotactic factors.

Effects of garlic Allium sativum on the immune responses of olive flounder Paralichthys olivaceus (마늘, Allium sativum이 넙치, Paralichthys olivaceus의 면역반응에 미치는 영향)

  • Lee, Jun-Hee;Woo, Sung-Ho;Eom, Yong-Hwan;Hwang, Bun-Ok;Kwon, Mun-Gyeong;Bang, Jong-Deuk;Park, Soo-Il
    • Journal of fish pathology
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    • v.23 no.1
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    • pp.69-83
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    • 2010
  • This study was aimed to investigate the effects of injection of garlic, Allium sativum, extract and immersion in garlic juice on the nonspecific immunity and the resistance against the artificial infection of Streptococcus iniae and Edwardsiella tarda of the olive flounder, Paralichthys olivaceus. The nonspecific immune mechanisms were assessed in terms of lysozyme activity, nitroblue-tetrazolium (NBT) assay and superoxide dismutase (SOD) activity etc. Relative percent survival (RPS) was assessed by the challenge with S. iniae BS10 or E. tarda KE-1. Almost of the garlic extract injected groups showed the enhanced level of the tested nonspecific immune factors. In the challenge test with S. iniae and E. tarda, RPS of 5% garlic extract pre-injected group was much higher than that of any other tested groups, respectively. Almost of the garlic juice immersion tested groups exhibited strengthened nonspecific immune defence factors, lysozyme activity, the number of lymphocytes and neutrophils, NBT reduction and SOD activity in kidney. In the challenge with S. iniae and E. tarda, RPS in the 0.25 g/L of garlic juice immersed group was much higher than any other tested groups, respectively. The results suggest that the garlic extract and juice would be effective to enhance the nonspecific immunity and protective ability of olive flounder against fish disease such as S. iniae and E. tarda.

Effect of Red ginseng extract on immune function of Israeli carp, Cyprinus carpio. (홍삼추출물이 이스라엘잉어의 면역반응에 미치는 영향)

  • Choi, Min-Soon;Park, Sung-Woo;Park, Kwan-Ha
    • Journal of fish pathology
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    • v.18 no.3
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    • pp.277-285
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    • 2005
  • Israeli carps were intraperitoneally injected with an aqueous extract of red ginseng (RG) at five different concentrations (0, 1, 10, 25 and 50 mg/kg of body weight). The fish were sacrificed after 3 days and measured the neutrophil activity and plasma components. The activity of lysozyme, Ht, TIC and NBT reduction were significantly enhanced in the concentrations of more than 10 mg/kg. Further attempts were made to measure the effects of RG on leucocyte function at five different concentrations (1-100 ${\mu}g/m{\ell}$), in vitro. The proliferation of lymphocytes co-incubated with phytohaemagglutin (PHA) and concanavalin A (Con A) was significantly increased at 10 ${\mu}g/m{\ell}$ concentration, but suppressed at the highest concentration (100 ${\mu}g/m{\ell}$). RG alone or with lipopolysaccharide (LPS) had no effect on proliferation at any concentrations tested. Migration and reactive oxygen intermediates (ROI) production of leucocytes were enhanced in all concentration higher than 10 ${\mu}g/m{\ell}$. These results suggest that the RG activates the leucocyte function and may be able to be used as a stimulant of fish innate defences.

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.

Characteristics of Photosynthetic Electron Transport Activity in Isolated Chloroplast of Korean Ginseng and Radish (인삼과 무 엽록체의 광합성 전자전달 활성)

  • 김갑식
    • Journal of Plant Biology
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    • v.33 no.2
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    • pp.111-118
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    • 1990
  • In order to characterize the chloroplasts of Korean ginseng as a semi-shade plant and radish as a sun plant, effects of growth light intensity on photosynthetic electron transport (PS) activity in chloroplasts and superoxide (O2.-) production in thylakoid membrane by irradiation were investigated. High-light chloroplasts of both plants showed higher PS activities than those grown under ow growth light intensity. High PS II and low PS I activities in ginseng chloroplasts (ratio of PS II/PS I : 1.1) were observed, but radish chloroplasts showed low PS II and high PS I activities (ratio of PS II/PS I : 0.3). PS II activity of both plants was little affected by temperature in range of 15-35$^{\circ}C$. Activities of whole -chain (PS II+I) in ginseng and PS I in radish were increased at high temperature (4$0^{\circ}C$). Preincubation of chloroplasts at 4$0^{\circ}C$ during 30 min, as a mild heat stress, caused rapid decrease in PS II and PS II+I activities of both plants. However PS I activity was not decreased in ginseng and rather increased in radish. O2.- production (NBT reduction) in Mehler reaction in the thylakoid membrane was inhibited by DCMU in both plants. DMBIB inhibited O2.- production in ginseng, but radish was insensitive to DMBIB. Electron flow system in ginseng thylakoid membrane was more susceptible to damage of photooxidation than that of radish.

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Analysis of the Biological Function of ELDF15 Using an Antisense Recombinant Expression Vector

  • Liu, Yan;Wang, Long;Wang, Zi-Jun
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.21
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    • pp.9131-9136
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    • 2014
  • ELDF15, homologous with AT2 receptor-interaction protein 1 (ATIP1), may play an important role in cell differentiation, proliferation, and carcinogenesis. We aimed to understand the biological function of ELDF15 via construction and transfection of a recombinant expression vector containing antisense ELDF15. Recombinant expression vectors were successfully constructed and transfected into K562 cells. A stable transfectant, known as pXJ41-asELDF15, stably produced antisense ELDF15. Compared with K562 and K562-zeo cells, K562-pXJ41-asELDF15 cells showed inhibition of cell proliferation. RT-PCR analysis showed that the expression and protein level of ELDF15 decreased significantly in K562 cells transfected with pXJ41-asELDF15. Expression of hemoglobin increased in K562 cells transfected with pXJ41-asELDF15 by benzidine staining. increases NBT reduction activity in K562 cells transfected with pXJ41-asELDF15.Colony forming efficiency in two-layer soft agar was clearly inhibited as assessed by electron microscopy. These results suggest that ELDF15 plays a potential role in cell differentiation, proliferation and carcinogenesis.

Antioxidant activity of Cinchona officinalis stem bark extracts

  • MN, Ravishankara;Padh, Harish;M., Rajani
    • Advances in Traditional Medicine
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    • v.3 no.4
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    • pp.205-211
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    • 2003
  • Stem bark of Cinchona sp. (Rubiaceae) is one of the well known drugs for its therapeutic values in traditional as well as modern medicine. Even though a lot of work has been carried out on quinoline alkaloids of Cinchona, its phenolic constituents received very little attention. In the present study, we evaluated antioxidant properties of C. officinalis stem bark methanolic extract and water extract containing phenolic compounds (total phenolics 21.37, 5.18% w/w respectively in the two extracts) in different in vitro and ex vivo models viz., antiradical activity by DPPH reduction, superoxide radical scavenging activity in riboflavin/light/NBT system, nitric oxide radical scavenging activity in sodium nitroprusside/Greiss reagent system and inhibition of lipid peroxidation induced by iron-ADP-ascorbate in liver homogenate and haemolysis of erythrocytes induced by phenylhydrazine in erythrocyte membrane stabilization study. Both the extracts exhibited very good antioxidant activity in all the models tested. The phenolic compounds including tannins present in the stem bark seem to offer protection from the oxidative damage.