• Title/Summary/Keyword: tetrazolium viability test

Search Result 31, Processing Time 0.024 seconds

Macroalgae as the Source for Environmental Assessment

  • Luyen, Hai-Quoc;Meinita, Maria D.N.;Hong, Yong-Ki
    • Journal of Marine Bioscience and Biotechnology
    • /
    • v.2 no.2
    • /
    • pp.77-80
    • /
    • 2007
  • Macroalgal tissues can be used as indicating materials for environmental assessment using several algal biotechnology techniques. As bioassay test organisms, macroalgal tissues are required as an axenic state for suitable biological indicators. Callus formation and blade regeneration under suitable culture conditions are also useful for the tests. Quantitative method using tetrazolium chloride or $alamarBlue^{TM}$ is devised on a rapid assessment of the seaweed viability. The use of RT-PCR especially differential display technique should provide the means for the detection and isolation of the responding genes induced by the environmental stress. Seaweed thriving in more environmental changes might contain more diverse biologically active substances.

  • PDF

Study on non-destructive sorting technique for lettuce(Lactuca sativa L) seed using fourier transform near-Infrared spectrometer (FT-NIR을 이용한 상추(Lactuca sativa L) 종자의 비파괴 선별 기술에 관한 연구)

  • Ahn, Chi-Kook;Cho, Byoung-Kwan;Kang, Jum-Soon;Lee, Kang-Jin
    • Korean Journal of Agricultural Science
    • /
    • v.39 no.1
    • /
    • pp.111-116
    • /
    • 2012
  • Nondestructive evaluation of seed viability is one of the highly demanding technologies for seed production industry. Conventional seed sorting technologies, such as tetrazolium and standard germination test are destructive, time consuming, and labor intensive methods. Near infrared spectroscopy technique has shown good potential for nondestructive quality measurements for food and agricultural products. In this study, FT-NIR spectroscopy was used to classify normal and artificially aged lettuce seeds. The spectra with the range of 1100~2500 nm were scanned for lettuce seeds and analyzed using the principal component analysis(PCA) method. To classify viable seeds from nonviable seeds, a calibration modeling set was developed with a partial least square(PLS) method. The calibration model developed from PLS resulted in 98% classification accuracy with the Savitzky-Golay $1^{st}$ derivative preprocessing method. The prediction accuracy for the test data set was 93% with the MSC(Multiplicative Scatter Correction) preprocessing method. The results show that FT-NIR has good potential for discriminating non-viable lettuce seeds from viable ones.

Cytocompatibility and cell proliferation evaluation of calcium phosphate-based root canal sealers

  • Mestieri, Leticia Boldrin;Zaccara, Ivana Maria;Pinheiro, Lucas Siqueira;Barletta, Fernando Branco;Kopper, Patricia Maria Polli;Grecca, Fabiana Soares
    • Restorative Dentistry and Endodontics
    • /
    • v.45 no.1
    • /
    • pp.2.1-2.7
    • /
    • 2020
  • Objectives: This study aimed to evaluate the cell viability and migration of Endosequence Bioceramic Root Canal Sealer (BC Sealer) compared to MTA Fillapex and AH Plus. Materials and Methods: BC Sealer, MTA Fillapex, and AH Plus were placed in contact with culture medium to obtain sealers extracts in dilution 1:1, 1:2 and 1:4. 3T3 cells were plated and exposed to the extracts. Cell viability and migration were assessed by 3-(4,5-dimethylthiazoyl)-2,5-diphenyl-tetrazolium bromide (MTT) and Scratch assay, respectively. Data were analyzed by Kruskal-Wallis and Dunn's test (p < 0.05). Results: The MTT assay revealed greater cytotoxicity for AH Plus and MTA Fillapex at 1:1 dilution when compared to control (p < 0.05). At 1:2 and 1:4 dilutions, all sealers were similar to control (p > 0.05) and MTA Fillapex was more cytotoxic than BC Sealer (p < 0.05). Scratch assay demonstrated the continuous closure of the wound according to time. At 30 hours, the control group presented closure of the wound (p < 0.05). At 36 hours, only BC Sealer presented the closure when compared to AH Plus and MTA Fillapex (p < 0.05). At 42 hours, AH Plus and MTA Fillapex showed a wound healing (p > 0.05). Conclusions: All tested sealers demonstrated cell viability highlighting BC Sealer, which showed increased cell migration capacity suggesting that this sealer may achieve better tissue repair when compared to other tested sealers.

The venom of jellyfish, Chrysaora pacifica, induces neurotoxicity via activating Ca2+-mediated ROS signaling in HT-22 cells

  • Yang, Yoon-Sil;Kang, Young-Joon;Kim, Hye-Ji;Kim, Min-Soo;Jung, Sung-Cherl
    • Journal of Applied Biological Chemistry
    • /
    • v.62 no.4
    • /
    • pp.347-353
    • /
    • 2019
  • Stings of jellyfish, which frequently occur in a warm season, cause severe pain, inflammation and sometimes irreversible results such as the death. Harmful venoms from jellyfish, therefore, have been studied for finding the therapeutic agents to relieve pain or to neutralize toxic components. However, it is still unclear if and how jellyfish venom reveal neuronal toxicity even though pain induction seems to result from the activation of nociceptors such as nerve endings. In this study, using HT-22 cell line, we investigated neurotoxic effects of the venom of Chrysaora pacifica (CpV) which appears in South-East ocean of Korea. In 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay, CpV significantly reduced the viability of HT-22 cells in a dose-dependent manner. Additionally, in 2',7'-Dichlorofluorescin diacetate fluorescence test under the culture condition lacking dominant inflammatory factors, CpV remarkably increased the production of intracellular reactive oxygen species (ROS). Reduced responsive fluorescence to Rhodamine123 and increased expression of intracellular cytochrome c were also observed in HT-22 cells treated with CpV. These indicate that CpV-reduced viability of HT-22 cells may be due to the activation of apoptotic signalings mediated with oxidative stress and mitochondrial dysfunction. Furthermore, removing Ca2+ ion or adding N-acetyl-Lcystein remarkably blocked the CpV effect to reduce the viability of HT-22 cells. The findings in this study clearly demonstrate that CpV may activate Ca2+-mediated ROS signalings and mitochondrial dysfunction resulting in neuronal damage or death, and suggest that blocking Ca2+ pathway is a therapeutic approach to possibly block toxic effects of jellyfish venoms.

Effect of Seed Dehydration and Temperature during Cold-Stratification on the Seed Quality of Panax ginseng C. A. Meyer (인삼 종자의 생리적 휴면타파기간 중 건조처리 및 저장온도가 종자 건전성에 미치는 영향)

  • Suh, Su Jeoung;Jang, In Bae;Yu, Jin;Jang, In Bok;Park, Hong Woo;Seo, Tae Cheol;Kweon, Ki Bum
    • Korean Journal of Medicinal Crop Science
    • /
    • v.25 no.4
    • /
    • pp.209-216
    • /
    • 2017
  • Background: Dehisced ginseng seeds need to be stored at cold temperatures for around 3 months to break their physiological dormancy, and thus, to aid in gemination. In the presence of high moisture in such an environment, seed spoilage and pre-germination may lower seed quality and productivity. To improve seed quality during cold-stratification, the effects of seed dehydration and temperature were tested. Methods and Results: In early December, dehisced ginseng seeds were dehydrated at 4 different levels and stored at $2^{\circ}C$ $-2^{\circ}C$, and $-20^{\circ}C$ for 3 months. Germination was carried out on the filter papers moistened with distilled water; emergence of root, shoot, and seed spoilage were assessed. Seed viability was examined by the tetrazolium test. More than 90% of the seeds stored at $2^{\circ}C$ and $-2^{\circ}C$ without drying or endocarp dehydration germinated, but seeds that were dehydrated to have a moisture content (MC) below 31% showed poor germination and lost their viability. In addition, the seeds stored at $-20^{\circ}C$ failed to show effective germination. Conclusions: Seed storage after endocarp dehydration might help to improve seed quality and increase seedling's ability to stand during the spring-sowing of ginseng.

Preparation and Characterization of PLGA Scaffold Impregnated Keratin for Tissue Engineering Application (케라틴이 함유된 조직공학적 PLGA 지지체의 제조 및 특성 분석)

  • Oh, A-Young;Kim, Soon-Hee;Lee, Sang-Jin;Yoo, James J.;Dyke, Mark van;Rhee, John M.;Khang, Gil-Son
    • Polymer(Korea)
    • /
    • v.32 no.5
    • /
    • pp.403-408
    • /
    • 2008
  • Keratin is the major structural fibrous protein providing outer covering such as wool, hair, and nail. Keratin is useful as natural protein. We developed the keratin loaded poly(L-lactide-co-glycolide) (PLGA) scaffolds (keratin/PLGA) for the possibility of the application of the tissue engineering using bone marrow mesenchymal (BMSCs). Keratin/PLGA (contents 0%, 10%, 20% and 50% of PLGA weight) scaffolds were prepared by solvent casting/salt leaching method. We characterized porosity, wettability, and water uptake ability, DSC of keratin/PLGA scaffold. We seeded BMSCs isolated from the femurs of rat into the inner core of the hybrid scaffold. Celluar viability were assayed by 3- (4,5-dimethylthiazol-2-yl) -2,5-diphenyl-tetrazolium bromide (MTT) test. We confirmed that keratin/PLGA scaffold is hydrophilic by wettability, and water uptake ability measurement results. In MTT assay results, cell viability in scaffolds impregnated 10 and 20 wt% of keratin were higher than other scaffolds. In conclusion, we suggest that keratin/PLGA scaffold may be useful to tissue engineering using BMSCs.

In vitro germination of Gastrodia verrucosa Blume and Hetaeria sikokiana Tuyama treated by NaOCl (NaOCl 처리에 의한 한라천마(Gastrodia verrucosa Blume)와 애기천마(Hetaeria sikokiana Tuyama)의 기내발아)

  • Bae, Kee-Hwa;Ko, Myoung-Suk;Choi, Sun-A;Lee, Hak-Bong;Kim, Nam-Young;Song, Jae-Mo;Song, Gwan-Pil
    • Journal of Plant Biotechnology
    • /
    • v.39 no.3
    • /
    • pp.163-168
    • /
    • 2012
  • An optimization in vitro seed germination was established by using triphenol tetrazolium chloride (TTC) test, which has been known as two rare orchids (Gastrodia verrucosa Blume and Hetaeria sikokiana Tuyama) in Jeju island. We have established proper NaOCl treatment for in vitro germination of G. verrucosa and H. sikokiana through TTC test. In the case of H. sikokiana, seed viability through TTC test was high with 95% in control. However, NaOCl 1% treatment for 30 minutes showed the highest embryo swelling rate to seed viability. Likewise, swelling formation of embryos, diameter of embryos, protocorm formation and diameter of protocorms of G. verrucosa was 87%, $59{\mu}m$, 91% and $138{\mu}m$ through NaOCl 1% treatment for 30 minutes. This result will be applied on the basic information for improving in vitro seed germination rate of G. verrucosa and H. sikokiana.

Viability Assay of Seaweeds Responding to Mountain Fire-Related Pollutants (산불 관련 해양환경오염원들에 대한 해조류의 활력 측정)

  • KANG Se-Eun;JIN Long-Guo;CHOI Jae-Suk;CHO Ji-Young;SHIN Hyun-Woung;HONG Yong-Ki
    • Korean Journal of Fisheries and Aquatic Sciences
    • /
    • v.36 no.3
    • /
    • pp.225-229
    • /
    • 2003
  • Plant ash and soil drainage, derived by frequent mountain fires during winter, might cause biological contamination to seaweeds at seashore and river mouse area. To thalli of Ulva pertusa, maximum non-lethal concentration(MNLC), lethal concentration 50 $(LC_{50})$ and minimum lethal concentration (MLC) of pine needle ash were shown as 60, 350 and 550 mg/mL, respectively. The yellow loess and granite sand did not damage at concentrations of 20 and 200 mg/mL, respectively To thalli of Porphyra yezoensis, the MNLC, LC5O, MLC of pine needle ash were shown as 0.08, 0.4 and 1.0 mg/mL, respectively. Effects of yellow loess and granite sand were approximately 1/2 and 1/10 of the ash. To thalli of Undaria pinnatifida, the pine needle ash, yellow loess and granite sand did not damage at the concentration range of 20 to 40 mg/mL. Change of pigments $(chlorophyll\;\alpha,\;lutein,\;\beta-carotene,\;phycoerthrin)$ was also determined at the MNLC, $LC-{50}$ and MLC of pine needle ash. Among three seaweeds tested, P. yezoensis produced the most 2.7-fold of lutein and 2.3-fold of $\beta-carotene$ at $LC-{50}$ of the ash. Thus the P. yezoensis, appeared as a sensitive indicator, could be used as one of test organisms for determination of the biological effect of pollutants contaminated in marine environment.

Antimicrobial Effect of Coptidis rhizome Extract against Mutans Streptococci and Periodontopathogens

  • Park, Soon-Nang;Lim, Yun Kyong;Kook, Joong-Ki
    • International Journal of Oral Biology
    • /
    • v.40 no.2
    • /
    • pp.79-83
    • /
    • 2015
  • The purpose of the study was to investigate the antimicrobial activity of the methanol extract of Coptidis rhizome against the type strains of cariogenic bacteria, Streptococcus mutans and Streptococcus sobrinus, and the periodontopathogens, Porphyromonas gingivalis, Prevotella intermedia, Treponema denticola and Aggregatibacter actinomycetemcomitans. The antimicrobial activities of the crude extract and the methanol extract fractions of Coptidis rhizome separated by silica gel chromatography were evaluated by determining the minimal bactericidal concentration (MBC) values, using the microdilution method. The cell viability test of the extracts of Coptidis rhizome on the KB cells was also studied by methyl thiazolyl tetrazolium (MTT) assay. Our results showed that the 11th fraction (F11) of the methanol extract had the greatest antimicrobial activity against the tested bacteria, with no associated cytotoxicity on the KB cells, upto a concentration of $50{\mu}g/ml$. These results suggest that the silica gel chromatography fraction F11 of the methanol extract of Coptidis rhizome, could be useful in the development of oral hygiene products as an antimicrobial agent for the prevention of dental caries and periodontal diseases.

CYTOTOXICITY OF RETROGRADE FILLING MATERIALS TESTED BY 51Cr RELEASE, MIT AND LD ACTIVITY (51Cr방출과 MTT 및 LD활성도를 이용한 역충전재의 세포독성에 관한 연구)

  • Choi, La-Young;Im, Mi-Kyung
    • Restorative Dentistry and Endodontics
    • /
    • v.19 no.2
    • /
    • pp.409-428
    • /
    • 1994
  • Endodontic surgery is performed when conventional endodontic therapy fails or is contraindicated. In such cases, retrograde filling materials including amalgam, composite resin, and various cements have been used. Biocompatibilty and margin sealing ability of retrograde filling materials are important for the long term success of endodontic surgery. In vitro cell culture is frequently used as the method of measuring the biocompatibilty of dental materials. The purpose of this study was to evaluate the cytotoxicity of six kinds of retrograde filling materials including newly developed light curing glass ionomer cements. Each material was mixed according to. the manufacture's instruction and evaluated as : freshly mixed, 24-hour after mixing, and 168-hour after mixing respectively. The elution solution was extracted after 24-hour contact with materials using media. Cytotoxicity was evaluated by direct contact, or elution contact. Test results of radiochromium($^{51}Cr$) release, cell viability using tetrazolium dye (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl dimethyltetrazolium bromide(MTT) test and lactate dehydrogenase(LD) of damaged L929 cells were analyzed. In the $^{51}Cr$ release of direct contact, all experimental retrograde filling materials except amalgam and glass ionomer cement showed increased cytotoxicity compared to control. In the $^{51}Cr$ release of elution solution, the released $^{51}Cr$ was so minimal that it was impossible. to evlauate the cytotoxicity exactly. The elution solutions of glass ionomer cement and IRM showed marked cytotoxicity in MTT test. LD enzyme activity was highest in tests of direct contact with composite, light curing composite, and light curing glass ionomer cement and IRM. Amalgam revealed least cytotoxicity while IRM showed cytotoxicity using all three methods. Composite, light curing composite and light curing glass iomomer cement were cytotoxic in the tests of $^{51}Cr$ release and LD activity. Glass ionomer cement showed cytotoxic effect only in the MTT method. From these results it is suggested that the standardization and optimization of cytotoxicity testing, especially using elution solutions, should be strongly advised.

  • PDF