• Title/Summary/Keyword: Neutral red

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Cytotoxicity of Crude Extracts of Rheum uudulatum L. with Human Kidney Epithelial Cell A498 (신장 상피세포주 A498을 이용한 대황(Rheum undulatum L.)추출물의 세포독성)

  • 나명석;진종언;조남철
    • The Korean Journal of Food And Nutrition
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    • v.13 no.5
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    • pp.460-464
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    • 2000
  • We have evaluated cytotoxic effects of four crude extracts of methylene chloride, ethyl acetate, butanol, water layer isolated Rheum undulatum in A498 cell line, human kidney epithelial cells. The cytotoxic evalutation was measured by colorimetric assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide(MTT) , neutral red(NR) and sulforhodamine protein B(SRB). These results obtained are as follows : MTT, NR and SRB quantities were significantly decreased in cultured A498 cells treated four crude extracts by increased concentrations. The cell cytotoxic effect of crude extracts of butanol layer was more stronger than others layer. The values of MTT$\sub$50/, NR$\sub$50/, SRB$\sub$50/ of crude extract of butanol layer and were measured both 0.63 mg/ml, 0.65 mg/ml, and 0.68 mg/ml, respectively and the values of water layer were 0.84 mg/ml, 0.82 mg/ml. and 0.80 mg/ml. respectively in cultured A498 cell line.

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Glutathione is the Major Defensive Mechanism against Oxidative Stress in Human Embryonic Stem Cell

  • 이건섭;이영재;김은영;박세필;임진호
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.78-78
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    • 2003
  • Embryonic stem (ES) cells, derived from preimplantation embryo, are able to differentiate into various types of cells consisting the whole body, or pluripotency. In contrast, terminally differentiated cells do not usually alter their nature but frequently die or transform if they are exposed to inappropriate external stimulations. In addition to the plasticity, ES cells are expected to be different from terminally differentiated cells in very many ways, such as patterns of gene expressions, ability and response of the cells in confronting environmental stimulations, metabolism, and growth rate. As a model system to differentiate these two types of cells, human ES cells (MB03) and terminally differentiated cells (HeLa), we examined the ability of these two types of cells in confronting a severe oxidative insult, that is $H_2O$$_2$. Approximately 1$\times$10$^4$ cells were plated in 96 well plate and serum starved for overnight. The conditioned cells were exposed to a various concentration of $H_2O$$_2$ fur 24 hrs and loaded with neutral red (50$\mu\textrm{g}$/ml) for 4 hrs, washed with PBS for 2 min three times, and entrapped dye was dissolved out using acetic ethanol. Cytotoxicity was determined by reading the amount of dye in the medium using microplate reader. equipped with 575 nm filter. Relative amount of the dye entrapped within MB03 or HeLa were not significantly different when cells were exposed up to 0.4 mM $H_2O$$_2$. However, this sharply decreased down to 0.12% in HeLa cells when the cells were exposed to 0.8 mM $H_2O$$_2$, while it was approximately 54% in MB03 suggesting that this concentration of $H_2O$$_2$ is the defensive threshold for HeLa cells. The resistance to oxidative stimulation reversed, however, when cells were co-treated with BSO (L-buthionine- 〔S, R〕-sulfoximine) which chelates intracellular GSH. This result suggests that cellular GSH is the major defensive mechanism of human ES cells. Induction of enzymes involved in GSH metabolism and type of cell death is currently being studied.

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Improvement of Ethanol Production by Electrochemical Redox Combination of Zymomonas mobilis and Saccharomyces cerevisiae

  • Jeon, Bo-Young;Park, Doo-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.20 no.1
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    • pp.94-100
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    • 2010
  • Zymomonas mobilis was immobilized in a modified graphite felt cathode with neutral red (NR-cathode) and Saccharomyces cerevisiae was cultivated on a platinum plate anode. An electrochemical redox reaction was induced by 3 volts of electric potential charged to the cathode and anode. The Z. mobilis produced 1.3-1.5 M of ethanol in the cathode compartment, whereas the S. cerevisiae produced 1.7-1.9 M in the anode compartment after 96 h. The ethanol produced by the Z. mobilis immobilized in the NR-cathode and S. cerevisiae cultivated on the platinum plate was 1.5-1.6 times higher than that produced under conventional conditions. The electrochemical oxidation potential inhibited Z. mobilis, but activated S. cerevisiae. The SDS-PAGE pattern of the total soluble proteins extracted from the Z. mobilis cultivated under the electrochemical oxidation conditions was gradually simplified in proportion to the potential intensity. Z. mobilis and S. cerevisiae were cultivated in the cathode and anode compartments, respectively, of an electrochemical redox combination system. The Z. mobilis culture cultivated in the cathode compartment for 24 h was continuously transferred to the S. cerevisiae culture in the anode compartment at a rate of 300 ml/day. Approx. 1.0-1.2 M of ethanol was produced by the Z. mobilis in the cathode compartment within 24 h, and an additional 0.8-0.9 M produced by the S. cerevisiae in the anode compartment within another 24 h. Thus, a total of 2.0-2.1 M of ethanol was produced by the electrochemical redox combination of Z. mobilis and S. cerevisiae within 48 h.

Cytotoxic Effects of Phytophenolics from Caesalpinia mimosoides Lamk on Cervical Carcinoma Cell Lines through an Apoptotic Pathway

  • Palasap, Adisak;Limpaiboon, Temduang;Boonsiri, Patcharee;Thapphasaraphong, Suthasinee;Daduang, Sakda;Suwannalert, Prasit;Daduang, Jureerut
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.449-454
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    • 2014
  • Background: Extracts of Caesalpinia mimosoides Lamk has been reported to possess anticancer effects, but the active ingredients and the anti-cancer mechanisms are still unknown. Materials and Methods: The effects of a C mimosoides Lamk extract on cell proliferation and apoptosis induction in human cervical carcinoma cell lines, namely HeLa, SiHa, and C33A, as well as in normal Vero cells, were investigated. Results: Treatment with 5 active fractions (F17-F21) of C mimosoides Lamk methanol extracts inhibited cell viability in a dose- and time-dependent manner. Neutral red assays indicated that treatment with F21 significantly decreased the viability of all cervical cancer cell lines compared to F21-treated normal cells. In addition, HPLC analysis revealed that F21 contained multiple phenolic compounds, namely gallic acid, caffeine, vanillic acid, ferulic acid and resveratrol. F21 had the lowest IC50 and, therefore, a much higher cytotoxicity than F20, F17, F19, and F18 by 20-, 25-, 46- and 47- fold, respectively. Analysis of activation of the apoptosis pathway using a caspase 3/7 activity assay revealed that F21 treatment resulted in a considerable increase in caspase activation in all cancer cell lines tested. At the same concentration of F21, HeLa cells had the highest caspase activity (6.5-fold) compared to the control. Conclusion: C mimosoides Lamk may be of value as an alternative therapeutic agent, especially in combination with other compounds offering possible of synergy of action. Moreover, HPV- and non-HPV-related cervical cancer cells may differ in their responses to treatment regimens.

Possible Anticancer Activity of Rosuvastatine, Doxazosin, Repaglinide and Oxcarbazepin

  • El Sharkawi, Fathia Zaky;El Shemy, Hany Abdelaziz;Khaled, Hussein Moustafa
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.1
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    • pp.199-203
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    • 2014
  • Background: Rosuvastatine, doxazosin, repaglinide and oxcarbazepin are therapeutic drugs available in the market for the treatment of different diseases. Potential to display antitumor activities has also been suggested. The aim of the current study was to evaluate their in vitro effects on some human transformed cell lines. Materials and Methods: Cytotoxicity of the four drugs was tested in MCF-7, HeLa and HepG2 cells by the neutral red assay method and also the effect of rosuvastatine and doxazosin against Ehrlich Ascities Carcinoma Cells (EACC) by trypan blue assay. Results: Rosuvastatine exerted the greatest cytotoxic effect against HepG2 cells with an $IC_{50}$ value of $58.7{\pm}69.3$; in contrast doxazosin showed least activity with $IC_{50}=104.4{\pm}115.7$. Repaglinide inhibited the growth of both HepG2 and HeLa cells with $IC_{50}$ values of $87.6{\pm}117.5$ and $89.3{\pm}119.5$, respectively. Oxcarbazepine showed a potent cytotoxicity against both HeLa ($IC_{50}=19.4{\pm}43.9$) and MCF7 cancer cells (($IC_{50}=22{\pm}35.7$).On the other hand the growth of EACC was completely inhibited by doxazosine (100% inhibition) while rosuvastatine had weak inhibitory activity (11.6%). Conclusions: The four tested drugs may have cytotoxic effects against hepatic, breast and cervical carcinoma cells; also doxazosine may inhibit the growth of endometrial cancer cells. Further investigations in animals are needed to confirm these results.

Seasonal Variation in the Dietary Fiber, Amino Acid and Fatty Acid Contents of Porphyra yezoensis (채취시기별 방사무늬김(Porphyra yezoensis)의 식이섬유, 아미노산 및 지방산 함량 변화)

  • Shin, Dong-Min;An, Se-Ra;In, Seo-Kyoung;Koo, Jae-Geun
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.46 no.4
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    • pp.337-342
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    • 2013
  • Porphyra yezoensis is potentially an excellent source of dietary fiber, amino acids, and eicosapentaenoic acid (EPA) because this red seaweed is available in large quantities and is rich in polysaccharides, proteins, and n-3 fatty acids. This study determined the insoluble dietary fiber (IDF), soluble dietary fiber (SDF), neutral detergent fiber (NDF), acid detergent fiber (ADF), amino acid, and fatty acid contents of P. yezoensis harvested monthly from November 2011 to March 2012. The total dietary fiber (TDF) and IDF contents ranged from 27.2-34.9% and 18.5-26.9%, respectively, and were greater in March than November. The SDF content ranged from 4.9-8.4% and did not differ significantly during growth. Galactose and 3,6-anhydro galactose were the major sugars in IDF and SDF. The higher levels of galactose and 3,6-anhydro galactose in IDF might be due to associated porphyran-type polysaccharides. Mannose and xylose were also major sugars in IDF. The total amino acid contents decreased gradually from November to March. The total amino acid composition of Porphyra was dominated by alanine, glutamic acid, arginine, and aspartic acid. No significant changes in the fatty acid profile were observed throughout the study period. The dominant fatty acid during all seasons was EPA, which comprised as much as 50% of the total fatty acid content.

Cellular Biomarker of Membrane Stability and Hydrolytic Enzyme Activity in the Hemocytes of Benzo(a)pyrene-exposed Pacific oyster, Crassostrea gigas

  • Jo Qtae;Choy Eun-Jung;Park Doo Won;Jee Young-Ju;Kim Sung Yeon;Kim Yoon
    • Fisheries and Aquatic Sciences
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    • v.5 no.4
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    • pp.263-270
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    • 2002
  • The Pacific oysters, Crassostrea gigas, were stressed with different concentrations of benzo(a) pyrene and depurated to determine the hemocyte lysosomal membrane stability and hydrolytic enzymatic activity as a biomarker candidate to the chemical, using NRR (neutral red retention) and API ZYM System, respectively. The membrane damage measured as NRR decrease was significant with the increase of chemical concentration and exposure time (P<0.05), providing a possible tool for biomarker. Interestingly, the control showed intrinsic stress probably due to captive life in the laboratory, and a recovering trend was also found during the depuration. The benzo(a)pyrene-exposed oysters showed increased enzyme activities in alkaline phosphatase, esterase (C4), acid phosphatase, naphthol-AS-BI-phospho­hydrolase, $\beta$-galactosidase, $\beta$-glucuronidase, and N-acetyl- $\beta$-glucosaminidase. Of them, only two enzymes, acid phosphatase and alkaline phosphatase, showed some potential available for the generation of enzymatic biomarker in the oyster. The results are suggestive of the potential availability of the cellular and enzymatic properties as a biomarker. However, considering that a robust biomarker should be insensitive to natural stress coming from normal physiological variation, but sensitive to pollutants, a concept of intrinsic stress the animal possesses should be taken into consideration. This reflects the necessity of further research on the intrinsic stress affecting the cellular and enzymatic properties of the chemical­stressed oysters prior to using the data as a biomarker.

A Study on the Antimicrobial Activity of Chitosan on the MRSA by Tube Dilution Technique and Agar Plate Smear Method (Tube Dilution Technique과 Ager Plate Smear Method에 의한 키토산의 MPSA 항미생물성)

  • Choi, Jeong-Im;Jeon, Dong-Won
    • Fashion & Textile Research Journal
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    • v.5 no.1
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    • pp.71-76
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    • 2003
  • Three different types of chitosan were prepared from red crab shells to study anti-microbial activity of chitosan on pathogenic bacteria, MRSA(Methicillin-resistant. Staphylococcus aureus), Water-insoluble chitosan, whose degree of deacetylation is kept over 90% and molecular weights are 20,000, 500,000, 150,000, 80,000, and 40,000, respectively. Water-soluble chitosan, whose degree of deacetylation is about 48% and molecular weights are 200,000 and 80,000. Water-soluble chitosan, whose degree of deacetylation is 82% and molecular weight is 3,900. The anti-microbial activities of three types of chitosan were investigated by Tube Dilution Technique(TDT) and Agar Plate Smear Method(APSM). And the following conclusions are made ; Chitosan having 5 different types of M.W chitosan (over 90% deacetylation) showed similar anti-microbial activities at over 0.05% concentration. Especially, chitosan having M.W 40,000 150,000 showed the excellent anti-microbial activity. The anti-microbial activity of chitosan was enhanced when the chitosan/acetic add solution was aged for 7days. The anti-microbial activity of chitosan was only shown at chitosan/acetic acid solution. The anti-microbial activity was not detected in chitosan solution dissolved in neutral pH water. Therefore, it can be concluded that the anti-microbial activity was due to NH3+ cationic ion of chitosan in acidic aqueous solution.

A Study on the Expression Types of Korean Beauty in Korean Fashion (한국 패션에 나타난 한국미의 표현유형에 관한 연구)

  • Choi, Hae-Joo
    • Journal of the Korean Society of Costume
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    • v.63 no.1
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    • pp.147-160
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    • 2013
  • Korean fashion designs became important items in Korea in the middle of the 1980s, and it has advanced into the world market in the early part of the 1990s. To achieve success in global fashion world, it is necessary to analyze Korean fashion design thoroughly and prepare strategies to develop designs with mainstream acceptance in the global fashion world. The purpose of this study is to analyze the aesthetic characteristics and Korean beauties of Korean fashion designs. Fashion photos of Korean fashion designs from 2006 S/S to 2012 F/W were analyzed. 357 designs from 608 designs of 4 representative Korean designers were examined and design characteristics, expression styles were studied. The major conclusions of the study are as follows : 1. H Line, slim, fitted silhouette and loose look were applied. 2. 5 major traditional colors, red, blue, yellow, white and black colors were used. Brown color, neutral color, golden color and beige color of the textile material's original color were used. Traditional textile materials like ramie fabric, satin and cotton, wool and metallic fabric were used. 3. Patterns of flower, traditional pattern and Korean letters were applied. Embroidery, patchwork and mother-of-pearl were decorated. 4. The three types of beauty were natural beauty, moderate beauty and decorative beauty. The types that were analyzed were realistic expression type, moderate expression type, image expression type and mixed expression type. To be accepted in global fashion world, Korean traditional design elements should be modified, broken down and reorganized so that Korean fashion design can be recreated.

Assessment of the Dermal and Ocular Irritation Potential of Lomefloxacin by Using In Vitro Methods

  • Ahn, Jun-Ho;Eum, Ki-Hwan;Lee, Mi-Chael
    • Toxicological Research
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    • v.26 no.1
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    • pp.9-14
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    • 2010
  • The evaluation of eye and skin irritation potential is essential to ensuring the safety of human in contact with a wide variety of substances. Despite this importance of irritation test, little is known with respect to the irritation potency of lomefloxacin, a fluoroquinolone antibiotic, which has been known to cause phototoxicity with an abnormal reaction of the skin. Thus, to investigate the tendency of lomefloxacin to cause eye and skin irritation, we carried out in vitro eye irritation test using Balb/c 3T3, and in vitro skin irritation test using $KeraSkin^{TM}$ human skin model system. 3T3 neutral red uptake assay has been proposed as a potential replacement alternative for the Draize Eye irritation test. In this study, the $IC_{50}$ value obtained for lomefloxacin was 375 ${\mu}g$. According to the classification model used for determining in vitro categories, lomefloxacin was classified as moderately irritant. For evaluation of skin irritation, engineered epidermal equivalents ($KeraSkin^{TM}$) were subjected to 10 and 25 mg of lomefloxacin for 15 minutes. Tissue damage was assessed by tissue viability evaluation, and by the release of a pro-inflammatory mediator, interleukin- 1${\alpha}$. Lomefloxacin increased the interleukin-1${\alpha}$ release after 15 minutes of exposure and 42 hours of post incubation, although no decrease in viability was observed. Therefore, lomefloxacin is considered to be moderately irritant to skin and eye.