• Title/Summary/Keyword: Fluorescence effect

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Inhibition of PI3K/AKT Signaling Pathway Enhances Cordycepin-Induced Apoptosis in Human Gastric Cancer Cells (인체위암 세포에서 PI3K/AKT 신호 전달계 차단에 의한 동충하초 유래 Cordycepin의 Apoptosis 유발 효과 증진)

  • Lee, Hye Hyeon;Jeong, Jin-Woo;Choi, Yung Hyun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.45 no.6
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    • pp.835-842
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    • 2016
  • The phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathway plays a crucial role in cancer occurrence by promoting cell proliferation and inhibiting apoptosis. In the present study, we evaluated the effect of a PI3K inhibitor, LY294002, on the chemosensitivity of gastric cancer cells to cordycepin, a predominant functional component of the fungus Cordyceps militaris, in AGS human gastric cancer cells and investigated possible underlying cellular mechanisms. Our results revealed that cordycepin inhibited viability of AGS cells in a concentration-dependent manner and induced apoptosis, as determined by apoptotic cell morphologies and fluorescence-activated cell sorting analysis associated with attenuated activation of the PI3K/Akt signaling pathway. Treatment with cordycepin in combination with a subtoxic concentration of LY294002 enhanced cordycepin-induced cytotoxicity and apoptotic potentials in AGS cells. Sensitization of LY294002 to cordycepin-induced apoptosis was accompanied by activation of caspases (caspases-3, -8, and -9) and was concomitant with poly(ADP-ribose) polymerase cleavage. Moreover, LY294002 up-regulated pro-apoptotic Bax and enhanced truncation of Bid in cordycepin-treated AGS cells, which was connected with increased loss of mitochondrial membrane potential and release of cytochrome c from mitochondria to the cytosol. Taken together, these results indicate that inhibition of the PI3K/Akt signaling pathway could augment cordycepin-induced apoptosis in human gastric cancer cells by up-regulating caspase activity through mitochondrial dysfunction.

Pepper Blight Disease Inhibition Metagenome Clone Screening Using Soil Metagenome Library (토양 Metagenome Library로부터 고추역병 저해 클론 탐색)

  • Park, Hae-Chul;Sung, So-Ra;Kim, Dong-Gwan;Koo, Bon-Sung;Jeong, Byeong-Moon;Kim, Jin-Heung;Yoon, Moon-Young
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.228-231
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    • 2009
  • We have purified Phytophthora capsici alpha and beta tubulin from Escherchia coli BL21(DE3). The recombinant alpha and beta tubulins were assembled into microtubule in vitro with specific conditions. The metagenome library was isolated from soil in the Mt. Yeo-Ki, Suwon, Korea and manufactured with the method mentioned in experiment contents for in vitro screening of microtubule assembly screening. FRET effect was used for microtubule assembly inhibitor screening with metagenome library. We got 2 metagenome clones from in vitro screening, and these 2 hit clones showed P. capsici growth inhibition activity on the growing pepper plants. These results suggest that new development of potent inhibitor for pepper blight disease and new approach to prevention of pepper blight disease.

Molecular Size Distribution and Spectroscopic Characterization of Humic and Fulvic Acids Extracted from Soils in Different Depth (깊이별 토양 부식산의 분자량분포 및 분광학적 특성 규명)

  • Shin, Hyun-Sang;Rhee, Dong-Sock;Chung, Kun-Ho;Lee, Chang-Woo
    • Analytical Science and Technology
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    • v.15 no.4
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    • pp.373-380
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    • 2002
  • Humic and fulvic acids present in soil of different depth were extracted and their characteristics were analyzed as a basic study to evaluate the effect of humic substances on the behaviour of radioactive elements deposited on soil. Molecular size distribution of the humic and fulvic acids was measured by stirred cell ultrafiltration technique and the structural informations were obtained from their UV-Vis., IR and synchronous fluorescence (SyF) spectral analysis. Main molecular size ranges of the soil 1) humic and fulvic acids were 30~100 kDa (46~56%) and 10~30 kDa (33~43%) respectively, and their overall molecular sizes were found to became smaller with increasing the soil depth. Absorptivities measured at 280 nm in the UV-Visible spectra of humic acids were 1.4~1.5 times higher than those of fulvic acids, and increased with increasing the soil depth. SyF spectral data showed two distinct peak components having maximum peak positions of 428 nm (type I) and 498 nm (type II) for the soil humic and fulvic acids. From the analysis of the peak components, it was found that humic molecules are mainly made up of aromatic compounds corresponding to longer wavelength (type II), and the molecular components increased with increasing the soil depth. Analysis of IR spectral data indicated that the humic molecules contain a higher relative concentration of carboxylic groups than those of fulvic molecules, and the carboxylic group contents are seen to increase as the soil depth increase.

Protective Effects of Bojungmyunyuk-dan in Cisplatin Treated Brain Cell Death (Cisplatin에 의한 뇌세포사멸에서 보중면역단의 방어효과)

  • Yoo Kyung Tae;Moon Seok Jae;Won Jin Hee;Kim Dong Woung;Lee Jong Deok;Won Kyoung Sook;Moon Goo
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.394-402
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    • 2003
  • This study was designed to investigate the protective effect of Bojungmyunyuk-dan(BJMY-Dan) on the cisplatin-induced cytotoxicity of primary rat astrocytes. BJMY-Dan is an oriental herbal prescription for its ability to recover protective effects against anti-cancer chemotherapies. After astrocytes were treated cisplatin, MTT assay was performed for cell viability test. To explore the mechanism of cytotoxicity, I used the several measures of apoptosis to determine whether this processes was involved in cisplatin-induced cell damage in astrocytes. Also, astrocytes were treated with BJMY-Dan and then, followed by the addition of cisplatin. Cisplatin decreased the viability of astrocytes in a dose and time-dependent manner. BJMY-Dan increased the viability of astrocytes treated cisplatin. Astrocytes treated cisplatin were revealed as apoptosis characterized by nuclear staining and flow cytometry. BJMY-Dan protected astrocytes from cisplatin-induced nuclear fragmentation and chromatin condensation. Also, caspase-3 and caspase-9 proteases were activated in astrocytes by cisplatin. BJMY-Dan inhibited the activation of caspase proteases in cisplatin-treated astrocytes. Cleavage of [poly(ADP-ribose) polymerase](PARP) was occurred at 12hr after treatment of cisplatin in astrocytes. BJMY-Dan recovered the cleavage of PARP in cisplatin-treated astrocytes. Also, BJMY-Dan inhibited the activation of pro-apoptotic factor, Bak by cisplatin. Lastly, astrocytes stained with JC-1 and Rhodamine 123 were photographed by fluorescence microscope to visualize changes of mitochondrial membrane permeability transition(MPT) during treatment with cisplatin for 24hr. BJMY-Dan recovered the change of MPT by cisplatin in astrocytes. According to above results, BJMY-Dan may protect astrocytes from cytotoxicity induced by chemotherapeutic agents, including cisplatin.

Increase of Larger-sized Pollen Number by Gametocide and Callus Induction in Anther Culture of Zoysia japonica Steud. (잔디의 약배양에 있어서 살정제 처리에 의한 Larger-sized Pollen의 발생빈도 증가 및 캘러스 유도)

  • Cho, Moon-Soo;Juang, Ue-Dong;Ye, Byong-Kwea;Ahn, Byung-Joon;Choi, Joon-Soo
    • Journal of Plant Biotechnology
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    • v.30 no.1
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    • pp.35-40
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    • 2003
  • In this study we investigated the effect of gametocides on the number of larger-sized pollen in anther, and also induced callus from the anther culture of Zoysia japonica Steud. Before culturing, we have observed pollens in anther through fluorescence and electron microscopes to know pollen dimorphism. There were two types of pollens observed. One type (30-36 ${\mu}{\textrm}{m}$ in diameter) consisted of vacuolated, larger-sized pollens and the other (15-20 ${\mu}{\textrm}{m}$ in diameter) smaller-sized ones with dense cytoplasm and plenty of amyloplasts. Within few hours, all the smaller-sized pollens were dead, while larger-sized ones were viable for one or two days. To induct larger-sized pollens, various gametocides were leaf-sprayed on three booting stages cultured under 4$0^{\circ}C$ /15$^{\circ}C$ (day/night) before anther culturing. Number of these larger pollens were few (less than 1%) in anther without spraying gametocides. GA$_3$increased the number of larger-sized pollens when applied at mid-booting stage. GA$_3$ with 50 mg/L treatment caused the highest percentage (25.4%) of the larger-sized pollen. Anthers with GA$_3$ treatment were only produced calli on AA medium (modified B$_{5}$+8.0 mg/L 2,4-D +0.2 mg/L kinetin), but callus formation was quite low (less than 1%).).

[ $2{\beta}$ ], $3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic Acid Induces the Apoptosis of Human Hepatoma HepG2 Cells ($2{\beta}$, $3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid처리에 의한 인간 간암세포주 HepG2의 apoptosis 유도)

  • Yoo, Ki-Hyun;Lee, Jong-Min;HwangBo, Jeon;Song, Myoung-Chong;Yang, Hye-Joung;Baek, Nam-In;Kim, Soung-Hoon;Kim, Dae-Keun;Kwon, Byoung-Mok;Park, Mi-Hyun;Chung, In-Sik
    • Applied Biological Chemistry
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    • v.49 no.4
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    • pp.270-275
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    • 2006
  • [ $2{\beta},\;3{\alpha}$ ], 23-trihydroxyrus-12-ene-28-oic acid was isolated from Trapa pseudoincisa S. et Z. It has a common structure of pentacyclic triterpenes and belongs to the amyrin ursolic acid group. The cytotoxic effect of this compound was investigated in human hepatoma cell line HepG2. $2{\beta},\;3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid showed dose-dependent cytotoxicity in HepG2 cells. Confocal microscopy data showed that green fluorescence was increased in $2{\beta},\;3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid treated-HepG2 cells in a time-dependent manner. $2{\beta},\;3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid also increased the sub-G1 cell population of HepG2 cells as well as ladder-like DNA fragmentation. Taken together, our results indicate that $2{\beta},\;3{\alpha}$, 23-trihydroxyrus-12-ene-28-oic acid induced apoptosis in HepG2 cells.

Immunological properties of the 30 kDa antigen of Toxoplasma gondii (단클론 항체를 이용하여 정제한 톡소포자충 30 kDa 항원의 면역학적 특성)

  • Lee, Yeong-Hwa;No, Tae-Jin;Sin, Dae-Hwan
    • Parasites, Hosts and Diseases
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    • v.35 no.1
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    • pp.55-62
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    • 1997
  • The molecular weight 30 kDa membrane protein of Toxoplusma Sondii (Toxoplasma 30 kDa) apparently conserved in most strains of T. gondii and sera of infected hosts. The present study aimed to elucidate Toxoplasmc 30 kDa as a useful diagnotic antigen for serodiagnisis of toxoplasmosis by ELISA and for induction of protective immunity. Murine spleen cells immunized with the membrane antigen of T. gondii were fused with mouse Sp2/0-Ag 14 myeloma cells. Out of 8 clones selected, five were IgG2b, the others belonged to IgG 1 and IgG2a. The 30 kDa antigen was distributed mainly on the surface membrane of tachyzoites by indirect fluorescence method. Murine peritoneal macrophages which were activated by 30 kDa antigen produced more amounts of NO2 compared with crude antigen-treated group, however there were no significant differences in toxoplamacidal activity between the two groups. Higher specificity of Toxoplosma 30 kDa antigen was recognized for serodiagnosis of toxoplasmosis than the crude antigen. From these results, ToxopLasmo 30 kDa antigen enhances the cytotoxic effect of macrophages as well as a more reliable means for the serodiagnosis of toxoplasmosis by ELISA. Key words: Toxoplosma gondii, 30 kDa antigen (p30), mouse, serodiagnosis, macrophage, cytotoxicity.

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Effect of Lactobacillus casei and a Fermented Milk on the Growth and Aflatoxin Production of Asperillus Parasiticus (유산균과 그 발효유가 Asperigllus parasiticus의 생육과 Aflatoxin 생성에 미치는 영향)

  • 김종규;이용욱
    • Journal of Food Hygiene and Safety
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    • v.13 no.2
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    • pp.164-170
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    • 1998
  • In this study a commercial fermented milk produced in Korea and a Lactobacillus strain used for the product (L. casei) were found to affect mold growth and inhibit aflatoxin production by Aspergillus parasiticus ATCC 15517. Aflatoxins were determined using an HPLC system that consisted of a $C_{18}$ column and a fluorescence detector. When the fermented milk was added to the yeast-extract broth the levels of aflatoxin $B_{1}\;and\;G_{1}$ significantly decreased by 48.6~58.1% and 29.8~34.2%, respectively (p$B_{1}\;and\;G_{1}$ were found in comparison with the control (monoculture). L. casei was found to be very inhibitory to the growth of A. parasiticus for 5 days, but no significant difference of mycelial weight was observed between the mixed culture and control at the end of incubation. The pH values of the culture broth in mixed culture were observed to be significantly lower than those in monoculture (p

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Expression and Role of Trypsin-Like Enzyme Involved in Hatching of Preimplantation Mouse Embryos (생쥐 배아의 부화에 관여하는 Trypsin 유사 효소의 발현과 역할)

  • Kim, Soo-Kyung;Kang, Hee-Kyoo;Jun, Jin-Hyun;Choi, Kyoo-Wan;Kim, Moon-Kyoo
    • Development and Reproduction
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    • v.5 no.1
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    • pp.17-21
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    • 2001
  • This study was conducted to investigate the expression pattern of Trypsin-like enzyme and the effect of a trypsin inhibitor(benzimidine) on hatching process during in-vitro culture of mouse preimplantation embryos. The Trypsin-like enzyme was identified by rhodamine-conjugated Trypsin substrate probe. The expression of trypsin-like enzyme was firstly detected at the late morula stage, and the enzyme was uniformly localized in the trophectoderm of late blastocysts. Especially, intense fluorescence was observed in the blebbing area of hatching blastocysts. Bisbenzamidine, contained in culture media, did not alter embryonic development from 4-cell stage to the expanded blastocyst but decrease the hatching rate in ImM concentration (15.8% vs 89.7%, p<0.02). In the treatment of bisbenzimidine (5mM) for 12 hours according to the embryonic stage of mouse, the hatching rate of control (83.0%) and treatment in late blastocysts (8.7%) were significantly (p<0.01) different. From these results, we suggested that the hatching enzyme having trypsin-like activity was localized from the late morula stage, and the hatching process by this enzyme was activated in the late blastocyst stage of mouse embryos.

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Evaluation of Photochemical Reflectance Index (PRI) Response to Soybean Drought stress under Climate Change Conditions (기후변화 조건에서 콩 한발스트레스에 대한 광화학 반사 지수 반응 평가)

  • Sang, Wan-Gyu;Kim, Jun-Hwan;Shin, Pyeong;Baek, Jae-Kyeong;Lee, Yun-Ho;Cho, Jung-Il;Seo, Myung-Chul
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.21 no.4
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    • pp.261-268
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    • 2019
  • Climate change and drought stress are having profound impacts on crop growth and development by altering crop physiological processes including photosynthetic activity. But finding a rapid, efficient, and non-destructive method for estimating environmental stress responses in the leaf and canopy is still a difficult issue for remote sensing research. We compared the relationships between photochemical reflectance index(PRI) and various optical and experimental indices on soybean drought stress under climate change conditions. Canopy photosynthesis trait, biomass change, chlorophyll fluorescence(Fv/Fm), stomatal conductance showed significant correlations with midday PRI value across the drought stress period under various climate conditions. In high temperature treatment, PRI were more sensitive to enhanced drought stress, demonstrating the negative effect of the high temperature on the drought stress. But high CO2 concentration alleviated the midday depression of both photosynthesis and PRI. Although air temperature and CO2 concentration could affect PRI interpretation and assessment of canopy radiation use efficiency(RUE), PRI was significantly correlated with canopy RUE both under climate change and drought stress conditions, indicating the applicability of PRI for tracking the drought stress responses in soybean. However, it is necessary to develop an integrated model for stress diagnosis using PRI at canopy level by minimizing the influence of physical and physiological factors on PRI and incorporating the effects of other vegetation indices.