• Environmental Analysis Health and Toxicology
• /
• 제14권1_2호
• /
• pp.1-12
• /
• 1999

Recently, several new methods for the detection of genetic damages in vitro and in vivo based on molecular biological techniques were introduced according to the rapid progress in toxicology combined with cellular and molecular biology. Among these methods, mouse lymphoma thymidine kanase (tk) gene forward mutation assay, single cell gel electrophoresis (comet assay) and transgenic animal and cell line model as a target gene of lac I (Big Blue) and lac Z (Muta Mouse) gene mutation are newly introduced based on molecular toxicological approaches. The mouse lymphoma tk$\^$+/-/ gene assay (MOLY) using L5178Y tk$\^$+/-/ mouse lymphoma cell line is one of the mammalian forward mutation assays, and has many advantages and more sensitive than hprt assay. The target gene of MOLY is a heterozygous tk$\^$+/-/ gene located in 11 chromosome, so it is able to detect the wide range of genetic changes like point mutation, deletion, rearrangement, and mitotic recombination within tk gene or deletion of entire chromosome 11. The comet assay is a rapid, simple, visual and sensitive technique for measuring and analysing DNA breakages in mammalian cells, Also, transgenic animal and cell line models, which have exogenous DNA incorporated into their genome, carry recoverable shuttle vector containing reporter genes to assess endogenous effects or alteration in specific genes related to disease process, are powerful tools to study the mechanism of mutation in vivo and in vitro, respectively. Also in vivo acridine orange supravital staining micronucleus assay by using mouse peripheral reticulocytes was introduced as an alternative of bone marrow micronucleus assay. In this respect, there was an International workshop on genotoxicity procedure (IWGTP) supported by OECD and EMS (Environmental Mutagen Society) at Washington D. C. in March 25-26, 1999. The objective of IWGTP is to harmonize the testing procedures internationally, and to extend to finalization of OECD guideline, and to the agreement of new guidelines under the International Conference of Harmonization (ICH) for these methods mentioned above. Therefore, we introduce and review the principle, detailed procedure, and application of MOLY, comet assay, transgenic mutagenesis assay and supravital staining micronucleus assay.

• Molecular & Cellular Toxicology
• /
• 제1권3호
• /
• pp.179-188
• /
• 2005

To develop the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, VI-3, VII-3, VIII-3, VII-20 and VII-20 (sodium salt) were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Single cell gel electrophoresis (Comet) assay, mouse lymphoma thymidine kinase ($tk^{+/-}$) gene assay (MOLY), chromosomal aberration assay in mammalian cells and Ames reverse mutation assay in bacterial system were used as simplified, inexpensive, short-term in vitro screening tests in our laboratory. Through the primary screening using the comet assay, we could choose the first candidates of sophoricoside derivatives with no genotoxic potentials as JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt). Also JSH-VII-3, VII-20 and VII-20 (sodium salt) are non-mutagenic in MOLY assay, while JSH-II-3 is mutagenic at high concentration with the presence of metabolic activation system in both comet assay and MOLY assay. The selected derivatives (JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt) are not mutagenic in S. typhimurium TA98 and TA100 strains both in the presence and absence of metabolic activation. From results of chromosomal aberration assay, 6 h treatment of JSH-VI-3, VII-3 and VII-20 (sodium salt) were not revealed clastogenicity both in the presence and absence of S-9 mixture. Therefore, we suggests that JSH-VI-3, VII-3, VII-20 and VII-20 (sodium salt), as the optimal candidates with both no genotoxic potential and IL-5 inhibitory effects must be chosen. To process the development into new anti-inflammatory drug of these derivatives, further investigation will need.

• 한국식품과학회지
• /
• 제31권5호
• /
• pp.1345-1348
• /
• 1999

고등어의 저장중 표피 오염 미생물 수를 신속하게 측정하여 신선도 판정을 위한 자료로 활용하고자 ATP bioluminescence assay에 의한 측정가능성을 파악하고자 하였다. 고등어를 $1^{\circ}C$에 저장하면서 표피를 swabbing 방법에 의해 미생물을 분리하여 총균수의 변화와 ATP bioluminescence assay에 의한 RLU 값을 측정하여 상관관계를 측정하여 본 결과 0.90 이상의 regression coefficient를 나타내어 유의적인 관계가 있음을 알 수 있었다. 따라서 ATP bioluminescence assay는 고등어 표피 오염 미생물의 신속 측정 방법으로 적합함을 알 수 있었으며 이 방법을 사용하였을 경우 기존의 48시간 이상 소요되는 전통적인 plate count 방법에 비하여 전처리를 포함하여 30분 이내에 결과를 알 수 있는 매우 신속하고 정확한 측정방법임을 알 수 있었다.

• 한국동물위생학회지
• /
• 제22권1호
• /
• pp.93-101
• /
• 1999

This test process on screening method for the detection of residual antibiotics in milk is simple, economic, sensitive to residual antibiotics and was given approval international organs. Thus, this study was carried out the comparison of Disk assay method and TTC-II method for sensitivity and minimum detectable range of antibiotics in raw milk. The results of this study was summarized as follows ; 1. The number of samples requested for treatment of mastitis was 198 samples. Comparison or analytical results among the methods of TTC-II, disk assay and Delve sp was that TTC-II 37 samples(18.6% ), Disk assay 125samples(63.1%), Delve SP 130 samples(65.7% ) reacted positively. Conformity rate of Delve SP and Disk assay was 70%. 2. Detectable limits of disk assay method in some antibiotics were more sensitive than those of official method(0.05-0.0025ppm in the $\beta$-lactams, 1ppm in two aminoglycoside, 0.2 ppm in one tetracycline, similar in one macrolide) 3. For sensitivity of residual sulfonamides TTC-II was much more sensitive than disk assay. Detectable limits of sulfamethazine and sulfadimethoxine were 30 to 50ppm levels. 4. The best medium preservation period is 1-2 days. 5. Concentration of brome cresol purple related to resistance for B stearothermophilus culture was 24ppm/ml. These results show that disk assay method for screening detection of antibiotics residuces in milk is worthy of use.

• 대한화장품학회지
• /
• 제15권1호
• /
• pp.37-50
• /
• 1989

The in Vitro chemosensitivity of fibroblast cell strains was determined using a semiautomated tetrazolium-based colorimetric assay(MTT assay) to 16 cosmetic materials. This assay is useful method to evaluate toxic effects of the chemicals. From assay results, we determined that the preservatives are more toxic than moisteurizers. The chemicals in the same group have a different toxicity. That is, in preservatives, Germall -115 is more toxic than Danisol -M, -p, and in surfactant sodium laurel sulfate than Myrj 52, and in moisteurizers, 1, 3-butylene glycol is more safe than the others. When the results from this assay for preservatives were compared with patch test results, good correlation was observed. Forthemore, this assay method can be used together with Patch test for the evaluation of the chemical toxicity, particularly in cosmetic field.

• 대한의생명과학회지
• /
• 제14권2호
• /
• pp.115-118
• /
• 2008

Salmonella typhi is frequent causes of foodborne illness and its detection is important for monitoring disease progression. In this study, by using general PCR and novel LAMP (Loop Mediated Isothermal Amplification) assay, we evaluated the usefulness of LAMP assay for detection of Salmonella typhi. In this LAMP assay, forward inner primer (FIP) and back inner primer (BIP) was specially designed for recognizing target invA gene. Target DNA was amplified and visualized as ladder-like pattern of bands on agarose gel within 60 min under isothermal conditions at $65^{\circ}C$. When the sensitivity and reproducibility of LAMP were compared to general PCR, there was no difference of reproducibility but sensitivity of LAMP assay was more efficient than PCR (the detection limit of LAMP assay was 30 fg, while the PCR assay was 3 pg). These results indicate that the LAMP assay is a potential and valuable means for detection of Salmonella typhi, especially for its rapidity, simplicity and low cost.

• Molecular & Cellular Toxicology
• /
• 제3권3호
• /
• pp.172-176
• /
• 2007

Butylated hydroxytoluene (BHT) is widely used antioxidant food additives. It has been extensively studied for potential toxicities. BHT appears adverse effects in liver and thyroid. In this study, we evaluated the genetic toxicity of BHT with more advanced methods, in vitro mouse lymphoma assay $tk^{+/-}$ gene assay (MLA) and in vivo mouse supravital micronucleus (MN) assay. BHT did not appear the significantly results in the absence and presence of metabolic activation system with MLA. Also, in vivo testing of BHT yielded negative results with supravital MN assay. These results suggest that BHT itself was not generally considered genotoxic.

• Molecular & Cellular Toxicology
• /
• 제3권2호
• /
• pp.113-118
• /
• 2007

Chlorobenzenes due to their acute toxicity and the capability of bioaccumulating are of great health and environmental concern. Especially, 1, 2-dichlorobenzene (CAS No. 95-50-1) is used for organic synthesis, dye manufacture, as a solvent and for other applications in chemical industry. Adverse effects of 1, 2-dichlorobenzene includes increases in liver and kidney weights and hepatotoxicity. In this study, we evaluated the genetic toxicity of 1, 2-dichlorobenzene with more advanced methods, in vitro mouse lymphoma assay $tk^{+/-}$ gene assay (MLA) and in vitro mouse supravital micronucleus (MN) assay. 1, 2-Dichlorobenzene appeared the significantly positive results and the induction of large mutant colonies only in the presence of metabolic activation system with MLA. But in vitro testing of 1, 2-dichlorobenzene yielded negative results with supravital MN assay. These results suggest that 1, 2-dichlorobenzene may play a mutagen rather than clastogen in vitro mammalian system.

• 식물병연구
• /
• 제13권3호
• /
• pp.145-151
• /
• 2007

Xanthomonas axonopodis pv. glycines에 의해 발병되는 콩 불마름병은 한국에서 콩에 가장 많이 발생하는 중요한 세균성 병해 중 하나이다. 본 연구에서는 Xanthomonas axonopodis pv. glycines를 종자에서 검출하기 위해 PCR기법을 이용하였으며, 한국의 36개 주요 콩 품종의 종자 오염을 조사하였다. 그리고 병원균 검출과 동정을 위한 PCR assay와 dilution plating assay를 비교하였다. PCR assay를 이용하여 인공접종에 의한 이병종자와 자연감염된 이병종자로부터 병원균 검출을 확인하였다. PCR assay를 통한 이런 결과는 dilution plating assay와 비슷한 결과를 보여 주었으며 종자에서 병원균을 검출하는 다른 전통적인 방법보다 더 효과적인 방법으로 증명되었다. 36개 주요 콩 품종의 X. axonopodis pv. glycines에 의한 종자 전염을 확인한 결과 풍산나물콩, 만리콩, 태광콩, 대망콩, 아주까리콩에서 병원균이 검출되었다. 그러므로 PCR assay는 콩 종자에서 신속하고, 민감하게 X. axonopodis pv. glycines 특이적으로 검출할 수 있는 효과적인 방법으로 활용될 수 있을 것이다.

• 한국환경성돌연변이발암원학회지
• /
• 제22권1호
• /
• pp.1-11
• /
• 2002