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Development of a Test Method for the Evaluation of DNA Damage in Mouse Spermatogonial Stem Cells
Hye Lyun Jeon,Jung-Sun Yi,Tae Sung Kim,Youkyung Oh,Hye Jeong Lee,Minseong Lee,Jin Seok Bang,Kinarm Ko,Il Young Ahn,Kyungyuk Ko,Joohwan Kim,Hye-Kyung Park,Jong Kwon Lee,Soo Jung Sohn 한국독성학회 2017 Toxicological Research Vol.33 No.2
Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they are still in an early stage. Consequently, we aimed to develop alternative test methods in male animals using mouse spermatogonial stem cells (mSSCs). Here, we modified the OECD TG 489 and optimized the in vitro comet assay in our previous study. This study aimed to verify the validity of in vitro tests involving mSSCs by comparing their results with those of in vivo tests using C57BL/6 mice by gavage. We selected hydroxyurea (HU), which is known to chemically induce male reproductive toxicity. The 50% inhibitory concentration (IC50) value of HU was 0.9 mM, as determined by the MTT assay. In the in vitro comet assay, % tail DNA and Olive tail moment (OTM) after HU administration increased significantly, compared to the control. Annexin V, PI staining and TUNEL assays showed that HU caused apoptosis in mSSCs. In order to compare in vitro tests with in vivo tests, the same substances were administered to male C57BL/6 mice. Reproductive toxicity was observed at 25, 50, 100, and 200 mg/kg/day as measured by clinical measures of reduction in sperm motility and testicular weight. The comet assay, DCFH-DA assay, H&E staining, and TUNEL assay were also performed. The results of the test with C57BL/6 mice were similar to those with mSSCs for HU treatment. Finally, linear regression analysis showed a strong positive correlation between results of in vitro tests and those of in vivo. In conclusion, the present study is the first to demonstrate the effect of HU-induced DNA damage, ROS formation, and apoptosis in mSSCs. Further, the results of the current study suggest that mSSCs could be a useful model to predict male reproductive toxicity.
Development of a Test Method for the Evaluation of DNA Damage in Mouse Spermatogonial Stem Cells
Jeon, Hye Lyun,Yi, Jung-Sun,Kim, Tae Sung,Oh, Youkyung,Lee, Hye Jeong,Lee, Minseong,Bang, Jin Seok,Ko, Kinarm,Ahn, Il Young,Ko, Kyungyuk,Kim, Joohwan,Park, Hye-Kyung,Lee, Jong Kwon,Sohn, Soo Jung Korean Society of ToxicologyKorea Environmental Mu 2017 Toxicological Research Vol.33 No.2
Although alternative test methods based on the 3Rs (Replacement, Reduction, Refinement) are being developed to replace animal testing in reproductive and developmental toxicology, they are still in an early stage. Consequently, we aimed to develop alternative test methods in male animals using mouse spermatogonial stem cells (mSSCs). Here, we modified the OECD TG 489 and optimized the in vitro comet assay in our previous study. This study aimed to verify the validity of in vitro tests involving mSSCs by comparing their results with those of in vivo tests using C57BL/6 mice by gavage. We selected hydroxyurea (HU), which is known to chemically induce male reproductive toxicity. The 50% inhibitory concentration ($IC_{50}$) value of HU was 0.9 mM, as determined by the MTT assay. In the in vitro comet assay, % tail DNA and Olive tail moment (OTM) after HU administration increased significantly, compared to the control. Annexin V, PI staining and TUNEL assays showed that HU caused apoptosis in mSSCs. In order to compare in vitro tests with in vivo tests, the same substances were administered to male C57BL/6 mice. Reproductive toxicity was observed at 25, 50, 100, and 200 mg/kg/day as measured by clinical measures of reduction in sperm motility and testicular weight. The comet assay, DCFH-DA assay, H&E staining, and TUNEL assay were also performed. The results of the test with C57BL/6 mice were similar to those with mSSCs for HU treatment. Finally, linear regression analysis showed a strong positive correlation between results of in vitro tests and those of in vivo. In conclusion, the present study is the first to demonstrate the effect of HU-induced DNA damage, ROS formation, and apoptosis in mSSCs. Further, the results of the current study suggest that mSSCs could be a useful model to predict male reproductive toxicity.
두 가지 단계적 접근법을 이용한 30종 화학물질의 안자극성 평가
전혜련 ( Hye Lyun Jeon ),조아랑 ( Ah Rang Cho ),고경육 ( Kyung Yuk Ko ),김주환 ( Joohwan Kim ),정미경 ( Mi Kyung Jeong ),박교현 ( Kyo-hyun Park ),김배환 ( Bae-hwan Kim ),오원준 ( Won Jun Oh ),이종권 ( Jong Kwon Lee ),박기숙 ( Ki S 한국동물실험대체법학회 2020 동물실험대체법학회지 Vol.14 No.1
Various alternative test methods are being developed to replace in vivo Draize rabbit test that evaluates eye irritation. However, a single alternative method has difficulty to be applied in safety evaluation on substances, because it cannot fully replace the in vivo test by simulating only part of the in vivo system. For this reason, different studies using several alternative test methods and test results of literatures have been under way. Our previous study suggested effective tiered approaches using three tests among Short Time Exposure (STE), Bovine Corneal Opacity and Permeability (BCOP), Hen’s Egg Test-Chorioallantoic Membrane (HET-CAM), re-constructed human cornea epitheliums (RhCE) tests. This study aimed to evaluate eye irritation potential for 30 new test chemicals using two tiered approaches. According to the data generated by direct test performance and literature survey, the accuracy, sensitivity and specificity of the two tiered approaches were 83.3%∼86.7%, 93.3%, 73.3%∼ 80%, respectively. Furthermore, the accuracy, sensitivity, and specificity of the two tiered approaches by applying combined data against 30 test chemicals in our present study and 47 test chemicals in previous study were 90.9%∼92.2%, 95.9%∼98.0%, 78.6%∼85.7%. Consequently, the two tiered approaches may be used to identify between irritants and non-irritants to replace in vivo test.
이광호,Mi Hye Kim,Jae Young Song,Sam Churl Kim,Jeong Uck Park,Seung Chul Baik,Hyung Lyun Kang,Seong Gyu Park,Hyang Ran Hwang,Dong Won Bae,윤희상,Myung Je Cho,Gyung-Hyuck Ko,Woo Kon Lee 대한미생물학회 2005 Journal of Bacteriology and Virology Vol.34 No.4
Low-abundance cellular proteins normally invisible on the standard two-dimensional SDS-polyacrylamide gel electrophoresis (2-DE SDS-PAGE) map must be enriched appropriately in order to be visualized and identified in cells or tissues. We applied proteins of H. pylori strain 26695 to a immobilized heparin-affinity resin, which has an affinity for nucleic acid-binding proteins, protein biosynthesis factors, and growth factors. The whole cell extract of H. pylori strain 26695 was fractionated by the heparin-agarose chromatography, and was analyzed by 2-DE. The 2-DE SDS-PAGE displayed spots after silver staining, which were identified by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). Among the ca. 150 spots that were processed, 79 proteins representing 57 genes were identified. Eleven proteins were determined to be nucleic acid-associated. Eighteen proteins were newly identified in this study, including DNA topoisomerase I. These results may provide guidance for enriching low abundance proteins of H. pylori and contribute to the construction of a master protein map of H. pylori.
Ko, Kyung Yuk,Jeon, Hye Lyun,Kim, Joohwan,Kim, Tae Sung,Hong, Yoon-hee,Jeong, Mi Kyung,Park, Kyo-Hyun,Kim, Bae-Hwan,Park, Sera,Jang, Won-hee,Cho, Sun-A,An, Susun,Cho, Ah Rang,Yi, Jung-Sun,Kim, Ji-Youn Pergamon 2020 Toxicology in vitro Vol.63 No.-
<P><B>Abstract</B></P> <P>In order to overcome the limitations of single <I>in vitro</I> eye irritation tests, Integrated Approaches to Testing Assessment strategies have been suggested for evaluating eye irritation. This study developed two tiered approaches combining alternative test methods. They were designed in consideration of the solubility property of test chemicals and to use the RhCE tests at final steps. The tiered approach A is composed of the STE, BCOP, HET-CAM or RhCE tests, whereas the tiered approach B is designed to perform simultaneously two <I>in vitro</I> test methods at the first stage and the RhCE test at the final stage. The predictive capacity of the two tiered approaches was estimated using 47 chemicals. The accuracy, sensitivity, and specificity value of the tiered approach A were 95.7% (45/47), 100% (34/34), and 84.6% (11/13), respectively, whereas those of the tiered approach B were 95.7% (45/47), 97.1% (33/34), and 92.3% (12/13), respectively. The approach A and B were considered to be available methods for distinguishing test chemicals of Category 1 (all 73.3%) and No Category (84.6% and 92.3%), respectively. Especially, the approach B was considered as an efficient method as the Bottom-Up approach, because it predicted correctly test chemicals classified as No Category.</P> <P><B>Highlights</B></P> <P> <UL> <LI> The proposed A and B approaches seem to be efficient strategies for evaluating eye irritation potency as they had higher predictive capacity than that of stand-alone <I>in vitro</I> test method. </LI> <LI> The B approach is considered to be an useful method for Bottom-Up approach in eye irritation IATA. </LI> </UL> </P>
심리적 탄력성과 자기 통제력이 대학생의 자살생각에 미치는 영향
임숙빈 ( Sook Bin Im ),허명륜 ( Myoung Lyun Heo ),김세영 ( Se Young Kim ),안혜영 ( Hye Young Ahn ),강샛별 ( Saet Byul Kang ),김미송 ( Mi Song Kim ),문혜미 ( Hea Mee Mun ),박선하 ( Sun Ha Park ),원아랑 ( A Rang Won ),이지혜 ( Ji Hy 한국간호과학회 정신간호학회(구 대한간호학회정신간호학회) 2014 정신간호학회지 Vol.23 No.4
Purpose: The study was done to examine undergraduate student`` suicidal ideation and to identify influences of individual characteristics including psychological resilience and self-control on suicidal ideation. Methods: Data were collected from 400 students in 11 universities in D city from May 1 to June 30, 2011. Data were analyzed with t-test, one-way ANOVA and Pearson` correlation using SPSS 19.0. Results: Of the students, 11.3% reported suicidal ideation levels higher than average. Woman students and those with high career anxiety, perceived poor health condition and bad relationships showed higher points on suicidal ideation. Suicidal ideation was also negatively correlated with psychological resilience and self-control. Conclusion: Over 10.0% of students need careful attention on suicidal ideation, especially counselling service related to suicide prevention for students with poor health, high career anxiety, and bad relationships. Various programs should be developed to promote psychological resilience to improve health and relationships, as well as job guidance for students. It is also suggested the students`` experiences be examined to determine how they overcame suicidal ideation.