• Asian-Australasian Journal of Animal Sciences
• /
• v.21 no.4
• /
• pp.494-498
• /
• 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.

• Journal of Environmental Health Sciences
• /
• v.23 no.2
• /
• pp.98-105
• /
• 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.

• YAKHAK HOEJI
• /
• v.45 no.3
• /
• pp.310-319
• /
• 2001

To compare skin irritation and cytotoxicity of anti-wrinkle agents, we examined skin irritation of six anti-wrinkle agents (ascorbic acid, glycolic acid, all trans-retinoic acid, ginseng extract, retinol, EB) in New Zealand white rabbit. Cytotoxicity of these agents was determined by MTT [tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide] at multi-time points in cultured HaCaT cell, a human immortalized keratinocyte cell. We then analyzed correlation between skin irritation and cytotoxicity by spearman's rank correlation analysis. All trans-retinoic acid showed the highest primary irritation index (0.92) in skin irritation test. Being all the six agents not irritant, retinal showed the most cytotoxic agents. The correlation between skin irritation and cytotoxicity ($IC_{50}$/ at different time point was 0.814, 0.757, 0.814 and 0.7 at 3, 24, 48 and 72 h, respectively. We also fecund that IC$_{20}$ and IC$_{80}$ of these agents showed similar correlation with skin irritation. These results therefore demonstrated that there is close correlation between skin irritation and cytotoxicity $IC_{50}$/ value by MTT in HaCaT cell at early time points by anti-wrinkle agents or IC$_{20}$ value. $IC_{50}$/ at earily time point or IC$_{20}$ values may be reliable alternative determinant of skin irritation.n.

• The Journal of Korean Medicine
• /
• v.20 no.4
• /
• pp.82-92
• /
• 2000

The purpose of the present study was to explore the proper method for animal stroke model of Oriental medicine To this end, brain ischemia was induced by distal middle cerebral artery occlusion(dMCAO) and proximal middle cerebral artery occlusion(pMCAO) and evaluated with the method of Triphenyl Tetrazolium Chloride (TTC) staining and Swimming Behavior Test. Results demonstrated that first, infarct size and volume of pMCAO group were significantly bigger that those of dMCAO group. Second, analysis of swimming behavior test revealed that the percentage of left turning angles of pMCAO was significantly bigger than that of dMCAO. Third, during swimming behavior test, there were peculiar traces of small successive circles that represent motor dysfunction and conscious disturbance among dMCAO group. The results of the study thus indicate that non-invasive intraluminal method of pMCAO was the appropriate animal stroke model for Oriental medicine in the light of brain ischemia as hemiplesia and conscious disturbance.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.59 no.3
• /
• pp.341-349
• /
• 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.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.34 no.2
• /
• pp.134-141
• /
• 1989

In order to estimate field seedling emergence and grain yield by the information collected from various laboratory seed vigor tests, each two malting and naked barley cultivar seeds were artificially aged according to the standard aging treatment suggested by the Association of Official Seed Analysts. The seed vigor tests adopted were warm- and cold-germination test, tetrazolium vigor test and electrocon. ductivity test. Field emergence of malting barley (Y) was estimated by Y=-2.962+0.229X$_1$ (% warm germination) -0.001X$_2$ (vigor of warm germination test: WGT) +0.354X$_3$ (vigor of cold germination test: CT) -0.558X$_4$ (% cold germination). The multiple correlation coefficient indicated that % warm germination. was contributed 64 % of the variation in seedling emergence rate of malting barley. The vigor of warm and cold germination tests, and % cold germination contributed additional 4, 7, and 9%, respectively, upon addition of the variables into regression. For naked barley, the regression equation of emergence rate was less efficient(R$^2$=54%) than that of malting barley(R$^2$=84%). A model to predict grain yield by the results of various seed vigor tests was not evaluated for both malting and naked barlev.

• Biomolecules & Therapeutics
• /
• v.3 no.3
• /
• pp.242-244
• /
• 1995

In vitro cell culture system has been proposed as a promising alternative model to in vivo skin irritation test. These studies were performed to screen the cytotoxicity effects of surfactants using normal human skin fibroblasts. Cell membrane integrity assessed by the leakage of lactate dehydrogenase (LDH) and mitochondrial integrity by MTT [3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromides reduction test were affected in a dose dependent manner. The irritation potential of surfactants to human skin patch test, and the changes of capillary permeability by rabbit intradermal safety test were assessed as in vivo methods. Our results suggest that LDH leakage assay and MTT reduction test using cultured human fibroblasts could be predictive for the irritancy of various surfactants in human, and LDH assay is superior correlated with in vivo test (r=0.886) to MTT test with in vivotest (r=0.757).

• Toxicological Research
• /
• v.6 no.1
• /
• pp.29-40
• /
• 1990

Cytotoxic effects of cytosine arabinoside and vinblastine on cultured fibroblasts were determined by colorimetric assays of neutral red (NR) and tetrazolium MTT, and by mutagenicity tests . Cytosine arabinoside and vinblastine were highly toxic by showing that concentrations of NR-50 and MTT-50 of two drugs were lower than 100 ${\mu}$M. At mid-point cytotoxicityvalue of two drugs, frequencies of micronuclei and SCEs were very high and chromosome showed structural abnormalities. The sizes of micronuclei formed by vinblastine were larger than those induced by cytosine arabinoside. These results suggest that cytosine arabinoside and vinblastine have highy mutagenic and severe cytotoxic effects on the cultured mouse fibroblasts.

• Asian Journal of Turfgrass Science
• /
• v.2 no.1
• /
• pp.70-77
• /
• 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.

• Proceedings of the Korean Society of Crop Science Conference
• /
• 2017.06a
• /
• pp.136-136
• /
• 2017

In agriculture, the rice is the one of important things. Many farmers and scientists have long tried to increase the yield of rice. So many technologies have been developed these days. One type of technology has given rise to a host of concerns and questions, namely Genetically Modified Organisms (GMOs). The increasing cultivation of GM crops has raised a wide range of concerns with respect to food safety, environmental effects and socio-economic issues and now commercially planted on about 100 million hectares in some 22 developed and developing countries. The scientific evidence concerning the environmental and health impacts of GMOs is still emerging, but so far there is no conclusive information on the definitive negative impacts of GMOs on health or the environment. Nevertheless, public perceptions about GMOs in food and agriculture are divided with a tendency toward avoiding GM food and products in many developed and developing countries. Also Korea is one of that country and is not allow the GMOs now. So I studied whether these GMOs are actually dangerous for environment and whether there are differences in cultivar characteristics such as germination test with TTC tetrazolium, germination test in frozen soil and gene-flow test with glufosinate and strip-bar test. With these experiments, we evaluated the agricultural safety of GM rice and to identify and assess environmental risks.