• Title/Summary/Keyword: tetrazolium viability test

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Comparative Studies on Cotton Seed Germinability with Tetrazolium Viability Test and X-ray Contrast Methods

  • Na, Young-Wang;Shim, Sang-In;Chung, Jung-Sung;Rho, Il-Rae;Kim, Seok-Hyeon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.2
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    • pp.188-193
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    • 2014
  • Seed viability testing provides valuable information for assessing seed lot germinability. However, most testing methods require destruction of seed prior to test. Because the dissected seeds for viability test cannot be used further evaluation, the nondestructively X-ray photography technique that can be applied for the evaluation of seed quality has been developed. In order to know the validity and accuracy of X-ray photography technique in seed evaluation test that conducted to remove the abnormal seed from a seed lot, we have compared the results from tetrazolium viability test, germination test and X-ray contrast method in cotton. Metallic salts treatment increased the efficiency of X-ray photographic method by enhancing the penetration of X-ray in abnormal or damaged seeds rather than normal seeds that have strong and well-organized tissues in seed. Cotton seeds presoaked for 16 hr in distilled water followed by soaking into metallic salt solution (5% NaI in water) for 60 min were easily classified seeds into dead seed and viable seed based on the radiography images obtained by X-ray radiation. We concluded that soft X-ray photography was reliable to find out the various defective characters due to heat and mechanical damage of seeds.

Evaluation of Boar Sperm Viability by MTT Reduction Assay in Beltsville Thawing Solution Extender

  • Byuna, J.W.;Choo, S.H.;Kim, H.H.;Kim, Y.J.;Hwang, Y.J.;Kim, D.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.4
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    • pp.494-498
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    • 2008
  • MTT (3-(4, 5-dimethyl thiazol-2-yl)-2, 5-diphenyl tetrazolium bromide) reduction assay is a method that validates the viability of an active cell. Dehydrogenase in mitochondria converts yellow colored insoluble tetrazolium salt to purple colored water-soluble formazan. Sperm also have mitochondria in the midpiece, therefore sperm viability could be evaluated by MTT reduction assay. Several studies have already demonstrated the capability of application of the MTT reduction assay to sperm of several species in Hepes-BSA buffer. Because most liquid semen was diluted in extender like BTS (Beltsville Thawing Solution), Modena or Androhep when it is used or transferred, semen needed another dilution in Hepes-BSA buffer to assess sperm viability. In this study, we evaluated boar sperm viability especially in BTS extended semen and compared the efficiency of this test with eosin-nigrosin staining. We used the fresh BTS extended semen from a local A.I center. Semen sample was diluted to $3.0{\times}10^7$ sperms/ml in BTS. The rates of formazan production were measured in 96-well microtiter plates immediately and 1h after incubation at $17^{\circ}C$ using a spectrophotometer at wave length 560 nm. Simultaneously, split samples of the same semen were tested, using eosin-nigrosin staining to compare the efficiency of the MTT assay of sperm viability in BTS. The correlation between the results of these tests was calculated using Student-t test and ANOVA. The results revealed a strong correlation between the results of MTT reduction rate and the results that were simultaneously determined by eosin-nigrosin staining at 1 h. In conclusion, the MTT reduction test was an effective and simple method to validate sperm viability and it could be used as a simple tool to evaluate sperm viability in the local A.I center and laboratory.

Assessment of Seed Viability and Vigour in Neem (Azadirachta indica A. Juss.)

  • Kumar, Devendra
    • Journal of Forest and Environmental Science
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    • v.29 no.4
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    • pp.282-291
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    • 2013
  • Rapid loss in viability of neem (Azadirachta indica A. Juss.) seed is a major problem. Present effort was undertaken for developing a set pattern for assessing of viability and vigour in seed of various mother tree age of neem (Age I-06 years, Age II-15 years, Age III-25 years and Age IV->30 years old). Various viability test viz. triphenyle tetrazolium chloride test, electrical conductivity, excised embryo test, and germination test have been performed on seeds obtained from mother tree age classes. Inconsistency was observed with the TTC and EC test in germination of seed in laboratory as well as nursery. While various vigour tests viz. cold test, chemical stress test (methanol stress test), and accelerated ageing test alongwith ageing index, germination test (G%, MGT and GV) and various seedling growth parameters like seedling length (cm), number of leaves, collar diameter (cm), total biomass (g) alongwith mathematical indices i.e. vigour index, sturdiness quotient, volume index, quality index, root shoot ratio in nursery as well have been taken for study and showed better consistency. On the basis present study results of various viability and vigour test indicated that mother tree age class II performed better in comparison to others and it can be recommended for seed collection. Further it is also recommended that viability of neem seed may be assessed using various laboratory tests like excise embryo test and germination test (G%, MGT and GV) and vigour test may be taken preferably by cold germination test, chemical (methanol) stress test, accelerated ageing test in laboratory and germination alongwith various seedling growth parameters seedling length (cm), number of leaves, collar diameter (cm), total biomass (g) alongwith mathematical indices like Vigour Index, Sturdiness quotient, Volume Index, Quality index, root shoot ratio in nursery as discussed in this study.

Effects of NaOCl treatment on in vitro germination of seeds of a rare endemic plant, Oreorchis coreana Finet

  • Bae, Kee-Hwa;Ko, Myoung Suk;Lee, Mi Hyun;Kim, Nam Young;Song, Jae Mo;Song, Gwanpil
    • Journal of Plant Biotechnology
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    • v.40 no.1
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    • pp.43-48
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    • 2013
  • Oreorchis coreana Finet is threatened globally by over-collection from its natural habitats for horticultural purposes. Its rarity in nature makes this plant one of the most endangered species in Korea. In this study, we investigated the effects of sodium hypochlorite (NaOCl) on orchid seed viability and seed germination. An in vitro bioassay swelling test using immature seeds was compared with a standard chemical procedure using triphenyl tetrazolium chloride (TTC) to test seed viability. In general, the bioassay was more appropriate for estimating embryo viability after a prolonged pre-treatment (more than 1 h) in 1% NaOCl, a surface sterilant often used to enhance germination of seeds of terrestrial plants. Therefore, an efficient method for investigating in vitro swelling of immature seeds is urgently needed. We established a method for determining the viability and swelling of O. coreana seeds via in vitro examination of immature seeds. Treatment of immature seeds with 1% NaOCl for 10 min greatly enhanced the extent of swelling of immature zygote embryos when compared to untreated seeds. These data obtained here appear to be comparable to viability and swelling that occurs in O. coreana seeds via asymbiotic germination.

Cytoprotective Effect of Organic Solvents Extracts of Sophorae Radix in H9c2 Cells (심근세포에서 고삼 유기용매 추출물의 항독성 효과)

  • Kwon Kang Beom;Kim Eun Kyung;Lim Yang Eui;Song Yung Sun;Park Jong Ha;Moon Hyung Cheal;Ryu Do Gon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.3
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    • pp.754-758
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    • 2004
  • To test the cytoprotective effect of sophorae radix (SR) against hydrogen peroxide (H₂O₂)-induced cytotoxicity, we investigated the cell viability using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay in the presence of methylene chloride, n-butanol, ethyl acetate and water soluble fraction of SR water extracts in H9c2 cells. These results were obtained as followed; H₂O₂ decreased the cell viability of H9c2 cells in a dose dependent manner. Cells pretreated with SR water extracts were protected the H₂O₂-induced decrease of viability in H9c2 cells. Among organic solvents fractions of SR water extracts, ethyl acetate soluble fractions of SR protected the decrease of viability induced by H₂O₂ in H9c2 cells. These results suggest that ethyl acetate soluble fractions of SR water extracts is effective in the prevention of H₂O₂-induced cytotoxicity.

Application of Seed Vigor Test for Predicting Field Emergence in Azuki Bean (Vigna angularis Wight) (팥 포장출현력 예측을 위한 종자세 검사)

  • Jeong, Gwan-Seok;Na, Young-Wang;Shim, Sang-In;Kim, Seok-Hyeon
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.59 no.3
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    • pp.341-349
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    • 2014
  • Field emergence of Azuki bean is poor due to hard seed coat as compared to other legumes. In this study, an attempt was made to develop prediction method with regression analysis based on various seed vigor tests in laboratory for field emergence of azuki bean. Azuki bean seeds artificially aged to provide various levels of seed quality were evaluated by the standard germination test (SGT), cold germination test (CT), cool germination test (CGT), complex stressing vigor test (CSVT), tetrazolium(TZ) vigor test and electroconductivity test. The SGT was suitable for predicting the field emergence in the unaged high vigor seeds. The abnormal seedling percentage and shoot length in the CGT were highly correlated with field emergence of moderate vigor seeds artificially aged for 2 days. Electroconductivity, seed viability in the CSVT, and vigor and predicted germinability in the tetrazolium vigor test were also useful for predicting field emergence. Percent of ungerminated seed in the CSVT was correlated with field emergence in the low vigor seeds artificially aged for 4 days. In a stepwise multiple regression analysis, seed viability in the SGT, normal seedling percentage and dry matter weight in the CGT accounted for 86.9% of the predicted value of field emergence in azuki bean.

A Study on the Cytotoxic Effect of Heavy metals (Cd, Ni, Zn) on Cultured Mouse Fibroblast L929 Cell line (생쥐 배양섬유 모세포주 L929에 미치는 중금속(Cd, Ni, Zn)류의 세포독성에 관한 연구)

  • 이종빈;나명석;황영진;위성욱;최진희;김선희;유춘만;김재민
    • Journal of Environmental Health Sciences
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    • v.23 no.2
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    • pp.98-105
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    • 1997
  • The study on the cytotoxicity of heavy metals was carried out to evaluate the cytotoxic effect of those on mouse L929 fibroblast cell in 96-well microtiter plates. The cytotoxicity was assayed by the neutral red, tetrazolium MTT, total protein, micronuclei test. The cytotoxicity of the heavy metals by neutral red and tetrazolium MTT was showed in order, cadmium > zinc > nickel for the cationic metals tested. The effect of metal-metal interaction on the cytotoxicity showed a marked reduction of cadmium toxicity by zinc, to a lesser degree, by nickel. The amount of total protein in treated group added heavy metals was less than that of the control and treated cadmium alone was less than those of combination with nickel or zinc. At midpoint cytotoxicity values of heavy metals, the frequency of micronuclei on the cell treated heavy metals was more than that of control and treated cadmium alone was more than those of combination with nickel or zinc. From those results, it could be suggested that the heavy metals decreased the viability of mouse fibroblast L929 cells in a concentration-dependent manner and have cytogenic toxic effects, but mixed group decreased the cytotoxic and cytogenic toxicity on L929 cells.

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Studies on the Seed's Germinability test by the T.T.C. testing method. (T.T.C 검정방법에 의한 종자의 발아력검정에 관한 연구)

  • 전우방
    • Asian Journal of Turfgrass Science
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    • v.2 no.1
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    • pp.70-77
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    • 1988
  • In order to determine and to reduce the differences between biochemical test method for the seed viability with T.T.C. (2, 3, 5. Triphenyl tetrazolium Chloride) reagent and germinator method, the topographical diagram of red colored formazan was carried out in 15-16 parts differences. From this resulted information, the classification of the typical staining reaction given for 3 species were derived into 3-4 parts for the germinable in normal seedling, the same as the following results. 1. Corn (Gram inaceae) * Entier embryo stained in bright red color. * Both extremities of scutellum unstained * Both extremities of scuttlium, coleorhiza and non-critial portions of radicle unstained. 2. Soybean (Leguminosae) *Seed completely stained in red color. *Minor unstained areas on cotyledons. *Extreme tip of radicle unstained; minor unstained areas on cotyledons 3. Radish (Cruciferae) *Seed completely stained. *Minor unstained areas on cotyledons. *Outer cotyledon mostly unstained: inner cotyledon completely stained Extreme tip of radicle unstained: large portion of outer cotyledon unstained.

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Biocompatibility and Bioactivity of Four Different Root Canal Sealers in Osteoblastic Cell Line MC3T3-El

  • Jun, Nu-Ri;Lee, Sun-Kyung;Lee, Sang-Im
    • Journal of dental hygiene science
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    • v.21 no.4
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    • pp.243-250
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    • 2021
  • Background: Endodontic sealers or their toxic components may become inflamed and lead to delayed wound healing when in direct contact with periapical tissues over an extended period. Moreover, an overfilled sealer can directly interact with adjacent tissues and may cause immediate necrosis or further resorption. Therefore, the treatment outcome conceivably depends on the endodontic sealer's biocompatibility and osteogenic potential. This study aimed to evaluate the cell viability and osteogenic effects of four different sealers in osteoblastic cells. Methods: AH Plus (resin-based sealer), Pulp Canal Sealer EWT (zinc oxide-eugenol sealer), BioRoot RCS (calcium silicate-based sealer), and Well-Root ST (MTA-based calcium silicate sealer) were mixed strictly according to the manufacturer's instructions, and dilutions of sealer extracts (1/2, 1/5 and 1/10) were determined. Cell viability was measured using the water-soluble tetrazolium-8 (WST-8) assay. Differentiation was assessed by alkaline phosphatase (ALP) activity and mineralized nodule formation by Alizarin Red S staining. Results: The cell viability of the extracts derived from the sealers excluding Well-Root ST was concentration dependent, with sealer extracts having the least viability at a 1/2 dilution. At sealer extract dilution of 1/10, the test groups showed the same survival rate as that control group, with the exception of BioRoot RCS. Among all experimental groups, BioRoot RCS showed the highest cell viability after 48 hours. The ALP activity was significantly higher in a concentration-dependent manner. Furthemore, all four materials promoted ALP activity and mineralized nodule formation compared to the control at 1/10 dilutions. Conclusion: This is the first study to highlight the differences in biological activity of these four materials. These results suggest that the composition of root canal sealers appears to alter the form of biocompatibility and osteoblastic differentiation.

Assessment of different pretreatments to breakage dormancy and improve the seed germination in Elaeocarpus serratus L. - an underutilized multipurpose fruit tree from South India

  • Raji, R.;Siril, E.A.
    • Forest Science and Technology
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    • v.14 no.4
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    • pp.160-168
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    • 2018
  • The seeds of Elaeocarpus serratus, a tropical underutilized fruit tree are characterized by hard seed coat and consequent poor water uptake and low germination. To improve the regeneration through seeds, various parameters such as viability of seeds, water uptake, and effect of seed mass on germination and pretreatments were performed using a completely randomized design (CRD). Tetrazolium (TZ) test was conducted using fresh, mature seeds revealed $50{\pm}2.56%$ mean viability. Seeds of different weight classes showed similar pattern of water uptake and the saturation level was achieved at 60 hrs of soaking. Seeds belong to weight class 2.6-3.5g were germinated ($12.5{\pm}1.26%$) with $175{\pm}1.75days$ (d) of mean time taken for germination (MTG). Germination capacity of seeds varied significantly among different populations and Varkala population gave $12.5{\pm}1.1%$ germination with $174.6{\pm}2.5d$ MTG. Among various seed treatments, mechanical scarification was superior in germination and significant reduction in MTG ($p{\leq}0.05$). The mechanical scarification by complete removal of seed coat resulted in $49.2{\pm}1.52%$ germination within a short period of time ($9.52{\pm}0.89d$ MTG). However, the complete removal of seed coat without damaging to embryo is a difficult task. An alternate treatment (Mechanical scarification II) by making cracks on nut faces vertically followed by soaking in distilled water for 24 hrs gave $48.4{\pm}1.73%$ germination with significantly reduced MTG ($12.14{\pm}0.56d$) over unsoaked, untreated control ($6.5{\pm}1.84%$ germination and $197.18{\pm}1.79d$ MTG; $p{\leq}0.05$). This treatment (Mechanical scarification II) is therefore recommended for E. serratus seeds as it can adopt easily and can achieve 7 fold increases in germination over control. The recorded germination through mechanical scarification is in tune with realized viability percentage of the seeds.