• Asian-Australasian Journal of Animal Sciences
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• 제21권4호
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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.

• 한국환경보건학회지
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• 제23권2호
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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.

• 약학회지
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• 제45권3호
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• pp.310-319
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• 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.

• 대한한의학회지
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• 제20권4호
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• pp.82-92
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• 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.

• 한국작물학회지
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• 제59권3호
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• pp.341-349
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• 2014

경실로 인해 다른 두과작물에 비해 포장 입묘가 불량한 팥의 포장출현율의 효과적인 예측 방법을 개발하기 위해 인위로 노화 정도를 조절한 종자에 대해 실험실에서 몇 가지 종자세 검사를 실시하여 포장출현력과 비교하였다. 얻어진 종자세 지표들에 대해 포장출현력과의 상관분석을 통해 예측을 위한 효율적인 지표들을 도출하였다. 팥에서 종자세가 높을 경우에는 표준발아검사에서의 종자활력률이 포장출현력 예측에 효과적이며, 약간 노화된 종자의 경우에는 저온발아검사(CGT)에서의 비정상묘율과 배축의 길이는 포장출현력과 높은 상관관계를 보였으며, 전기전도도와 CSVT에서의 활력률과 테트라졸리움 검사에서 종자세와 예측발아율은 포장출현력과 5% 수준에서 상관관계가 인정되었다. 종자세가 낮은 종자에서는 CSVT에서 불발아종자율이 포장출현력과 5% 수준에서 상관을 보였다. 포장출현력 예측에 효과적인 몇 가지 종자세지수를 이용하여 stepwise multiple regression 분석을 실시한 결과 테트라졸리움(TZ) 검사에서의 예측발아율은 결정계수($R^2$)가 0.820으로 포장출현력 예측에 가장 효과적인 검사방법임을 알 수 있었다. 표준발아검사(SGT)에서의 종자활력률, 저온발아검사(CGT)에서의 정상묘율과 건물중을 회귀방정식에 추가함에 따라 86.9% 까지 포장출현력 예측효율을 증가시킬 수 있었다.

• 한국작물학회지
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• 제34권2호
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• pp.134-141
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• 1989

보리종실의 샐리적 특성을 포장출아력과 비교함으로써 포장유묘출아율과 수량의 예측에 적합한 검정방법을 구명하고자 맥주보리 2품종과 쌀보리 2품종을 인위노화처리시켜 고온발아검정(warm germination test), 저온발아검정(cold germination test), Tetrazolium 유모세검정(TZ vigor test), 전기전도도검정(electroconductivity test)의 결과를 포장출아율 및 수량과 비교 검토 하였다. 고온발아검정과 저온발아검정이 포장출아율 예측에 효과적인 검정법으로 나타났으며, 이들의 관계는 맥주보리에서는 Y(포장출아율)=-2.962+0.229X$_1$ (% warm germination) -0.001X$_2$ (vigor, WGT) +0.354X$_3$ (vigor, CT) -0.558X$_4$ (% cold germination)으로 나타났다. 다중화귀방정식의 결정계수는 맥주보리의 경우 고온발아검정에서의 발아율만으로 포장출아율을 64%예측할 수 있었다. 고온발아유묘세, 저온발아유묘세, 저온발아율은 각각 4, 7, 9%씩 결정계수를 높혔다. 네가지 검정치의 누가적 예측효율은 맥주보리에서 84%, 쌀보리 54%였다. 수량에서는 포장출아율에서 보다 맥주보리, 쌀보리 공히 예측효과가 낮게 나타났다.

• Biomolecules & Therapeutics
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• 제3권3호
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• pp.242-244
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• 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
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• 제6권1호
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• pp.29-40
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• 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.

• 아시안잔디학회지
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• 제2권1호
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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.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.136-136
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• 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.