• Title/Summary/Keyword: DNA damages

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Inhibitory Effects of Epigallocatechin Gallate on Apoptosis in Human Vascular Endothelial Cells (혈관내피세포의 세포사멸작용에 대한 (-)Epigallocatechin Gallate의 억제효과)

  • Choi, Yean-Jung;Choi, Jung-Suk;Lee, Se-Hee;Lee, Yong-Jin;Kang, Jung-Sook;Kang, Young-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.31 no.4
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    • pp.672-678
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    • 2002
  • Oxidative stress contributes to cellular injury following clinical and experimental ischemia/reperfusion scenarios. Oxidative injury can induce cellular and nuclear damages that result in apoptotic cell death. We tested the hypothesis that the catechin flavonoid of (-)epigallocatechin gallate, a green tea polyphenol, inhibits hydrogen peroxide ($H_2O$$_2$)-induced apoptosis in human umbilical vein endothelial cells. The effect of apigenin, a flavone found in citrus fruits, on apoptosis parameters was also examined. A 30 min pulse treatment with 0.25 mM $H_2O$$_2$ decreased endothelial cell viability within 24 hrs by > 30% ; this was associated with nuclear condensation and biochemical DNA damage consistent with programmed cell death. In the 0.25 mM $H_2O$$_2$apoptosis model, 50${\mu}{\textrm}{m}$ (-)epigallocatechin gallate markedly increased cell viability with a reduction in the nuclear condensation and DNA fragmentation. In contrast, equimicromolar apigenin increased cell loss with intense DNA laddering, positive nick-end labeling and Hoechst 33258 staining. Thus, polyphenolic (-)epigallocatechin gallate, but not apigenin flavone, qualify as an antioxidant in apoptosis models caused by oxidative stress. Further work is necessary for elucidating the anti-apoptotic mechanisms of polyphenolic catechins.

The Effects of Jihwangyeumja and GamiJihwangyeumja water extract on The Cultured Primary Hippocampal Cell Damaged by XO/HX (지황음자와 가미지황음자 유출액이 XO/HX로 손상된 배양 해마신경세포에 미치는 효과)

  • Lee Yong Geun;Kim Sang Ho;Min Sang Jun;Yang Hee Suk;Jang Hyun Ho;Kim Tae Hean;Kang Hyung Won;Lyu Yeoung Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.16 no.5
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    • pp.989-1000
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    • 2002
  • The purpose of this study is to examine the toxic effects caused by xanthine oxidase/hypoxanthine(XO/HX) and the effects of herbal extracts such as JHYJ and GJHYJ on the treatment of the toxic effects. For this purpose, experiments with the cultured hippocampal cells from new born mice were done. The results of these experiments were as follows. 1. XO/HX, a oxygen radical-generating system, decreased the survival rates of the cultured cells on XTT assay and INT assay, the amount of DNA syntheses, and the amount of neurofilaments, and increased the lipid peroxidation. 2. JHYJ and GJHYJ have the efficacy of increasing the survival rates of the cultured cells. 3. JHYJ and GJHYJ have the efficacy of increasing the amount of neurofilaments and of decreasing the lipid peroxidation. 4. JHYJ and GJHYJ have the efficacy of increasing the amount of DNA syntheses. From the above results, it is suggested that Jihwangyeumja and Gamijihwangyeumja have marked efficacy as a treatment for the damages caused by the XO/HX-mediated oxidative stress. And Jihwangyeumja and Gamijihwangyeumja are thought to have certain pharmacological effects. Further dinical study of this pharmacological effects of Jihwangyeumja and Gamijihwangyeumja should be complemented.

A Study on DNA Degeneration by Comet Assay & Pathological Observation for Mouse Which were Exposed HCN Gases from Fire (화재로 인한 HCN 가스에 노출된 마우스의 병리학적 관찰 및 단세포 전기영동법을 사용한 DNA 변성 추적에 관한 연구)

  • Cho, Nam-Wook;Oh, Eun-Ha;Hwang, Sung-Kwy
    • Journal of the Korean Institute of Gas
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    • v.16 no.6
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    • pp.7-16
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    • 2012
  • Combustion Toxic Effects among several factors of risk encountered during fire are important in the evacuation and final survival, and they are broader and fatal than the direct damages caused by flame. Most studies on fire toxicity until the present are limited to fatality, mainly deaths by fire through pathological research. In this study, it is conducted as a fundamental experiment to address 3 principles of animal experiment and to provide an alternative test to animal testing that is regulated by national building codes and it was conducted through approval by the animal testing ethics committee. Hence, in this study average time of activity stop was measured after directly inhaling toxic gases (HCN) to laboratory animals (mice) through gas toxicity test (KS F 2271) for major asphyxiating gases(HCN) which are produced during fire combustion. effects of Combustion toxic gases on body were quantitatively analyzed through changes in internal organs and hematological analysis, and electrophoresis of a single cell of these laboratory animals. Biological conclusion of combustion toxicology is drawn through approaches (pathological examination, blood test, blood biochemical test, electrophoresis analysis of single cell) which could not confirmed in existing gas toxicity test.

Recent Advanced Toxicological Methods for Environmental Hazardous Chemicals (환경 오염물질의 진보된 독성 평가 기법)

  • 류재천;최윤정;김연정;김형태;방형애;송윤선
    • Environmental Analysis Health and Toxicology
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    • v.14 no.1_2
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    • pp.1-12
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    • 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.

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Protective Effects of Boyanghwanoh-tang on Serum and Glucose Deprivation-induced Apoptosis of PC12 Cells (보양환오탕이 영양혈청결핍에 의한 PC12 세포의 고사에 미치는 영향)

  • 김종길;정승원;임준모;장호현;윤종민;이기상;문병순
    • The Journal of Korean Medicine
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    • v.24 no.2
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    • pp.179-192
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    • 2003
  • Objectives : Boyanghwanoh-tang (Buyanhaiwu-tang) has been used as a prescription for stroke, senile and vascular dementia, ischemic brain and heart damage in Oriental traditional medicine. However, there is little known about the mechanism by which the water extracts of Boyanghwanoh-tang (Buyanhaiwu-tang) rescue cells fromthese damages, and little is known about the protective mechanisms of Boyanghwanoh-tang (Buyanhaiwu-tang) on oxidative stress in neuronal cells. Therefore, we have investigated the role of Boyanghwanoh-tang (Buyanhaiwu-tang) on serum and glucose deprived apoptosis in PC12 cells. Methods : PC12 Cells have been used extensively as a model for studying the cellular and molecular effects of neuronal cells. The viability of cells was measured by MIT assay. We used DNA fragmentation and caspase 1, 2, 3, 6, 9-likeproteases activation assay. Transcriptional activation of NF-kB was assessed by using electrophoretic mobility shift assay. Results : Boyanghwanoh-tang (Buyanhaiwu-tang) rescued PC12 cells from apoptotic death by serum and glucose deprivation in a dose-dependent manner. The nuclear staining of PC12 cells clearly showed that Boyanghwanoh-tang (Buyanhaiwu-tang) attenuated nuclear condensation and fragmentation, which represent typical neuronal apoptotic characteristics. Boyanghwanoh-tang (Buyanhaiwu-tang) also prevents fragmentation of genomic DNA and activation of caspase 3-like protease in serum and glucose deprived PC12 cells. Furthermore, Boyanghwanoh-tang (Buyanhaiwu-tang) reduced the activation of NF-kB by serum and glucose-deprived apoptosis. Conclusions : These findings suggest that serum and glucose deprivation induces reduced glutathione (GSH) depletion, and consequently, apoptosis through endogenously produced reactive oxygen species in PC12 cells. Also, our data indicated that Boyanghwanoh-tang (Buyanhaiwu-tang) has protective effects against the serum and glucose deprived deaths of PC12 cells, which are mediated by the generation of GSH that, in turn, can reduce oxidative stress caused by reactive oxygen species (ROS) such as hydrogen peroxide.

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Cultural characteristics of fungal species associated with deterioration or foxing of paper and chemical removal (종이변색균류의 배양적 특성 및 화학적 방법에 의한 변색제거)

  • Jo, Seong-Eun;Kim, Yong-Tae;Jeong, So-Yeong;Jo, Byeong-Muk;Lee, Jong-Gyu
    • Proceedings of the Korea Technical Association of the Pulp and Paper Industry Conference
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    • 2009.04a
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    • pp.295-303
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    • 2009
  • The annals of Joseon dynasty, especially the volumes of King SeJong(1418-1450 A.D.), were heavily deteriorated by fungi. Investigations on the deteriorating and foxing fungi were carried out. Fungal structures on the beeswax, which were coated on the both side of Han-Ji, were suspected to be involved in the deterioration, and were observed by SEM. Isolation and culturing of these fungi were tried by scrubing swab samples and placing on the artificial media. Culture-independent approaches were used to identify the fungal strains associated with damages of beeswax and foxing of the paper by the analyses based on DNA sequences data from the specific ITS region of rDNA regions. In addition, well-known paper staining fungi(PSF), i.e., Aspergillus terreus var. terreus, Fusarium oxysporum, Chaetomium globosum, Cladosporium cladosporioides, and Alternaria solani, were compared in the mycelial growth and stain on beeswax and papers under different environmental conditions (temperature, light, moisture, etc). Fungal strains isolated from the air samples in the storage room and shelves were identified as Irpex sp., Arthrinium sacchari, Cladosporium tenuissimum, Aspergillus sclerotiorum, Sistotrema brinkmannii, and Hypoxylon bovei var. microsporum The isolated strains were compared in growth and stain patterns on beeswax and papers(Han-Ji, Hwa-Ji, and Yang-Ji) whether these can cause damage or foxing on the annals or not.

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Effect of Bombycis Corpus on Male Reproductive Dysfunction Caused by Aging (노화로 발생한 남성 생식기능 이상에 백강잠(白殭蠶)이 미치는 영향)

  • Kim, Hyeong-Jun;Ahn, Sang-Hyun;Ahn, Ha-Young;Park, Sun-Young
    • The Journal of Korean Obstetrics and Gynecology
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    • v.32 no.1
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    • pp.1-14
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    • 2019
  • Objectives: This study was designed to investigate the effect of Bombycis corpus on reproductive dysfunction caused by aging. Methods: The experimental group was divided into three groups: a control group consisting of 8-week-old male ICR mice without any treatment, An aging-elicited group (AE group) consisting of 50-week-old ICR male mice without any treatment, and a Bombycis corpus treatment group (BC group) consisting of 50-week-old ICR male mice with treatment Bombycis corpus extract (0.78 g/kg/day) for 6 months. After 6 months, histochemistry and immunohistochemistry of the testis were performed to investigate the effects of Bombycis corpus on the reproductive dysfunction caused by aging. Results: In the first step, Bombycis corpus increased spermatogenesis and distribution of sertoli cells in the seminiferous tubule, increased BrdU positive reaction in the spermatogonium at the basal part of the seminiferous tubule, and decreased the apoptosis of Sertoli cells in the seminiferous tubule. In addition, Bombycis corpus increased AR positive in Sertoli cells and $17{\beta}-HSD$ positive in leydig cells. Finally, Bombycis corpus decreased 8-OHdG positivite reaction in the spermatids of the seminiferous lumen, caspase-3 positivity in leydig cells, and HDAC1 positivite reaction in sertoli cells. Conclusions: These results suggest that Bombycis corpus increases spermatogenesis, decreases apoptosis of leydig cells and Sertoli cells, increases the production and action of testosterone in the testis, and inhibits DNA damages and DNA transcripts decrease in the testis, Thereby improving reproductive dysfunction caused by aging.

Protective Effects of Omijatang on Oxidative Stress-Induced Apoptosis of H9c2 Cardiomyoblast Cells (오미자탕(五味子湯)이 산화적 손상으로 유발된 세포고사에 미치는 영향)

  • Choi, Jin-Young;Shin, Sun-Ho;Lee, Yun-Jae
    • The Journal of Internal Korean Medicine
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    • v.26 no.2
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    • pp.420-430
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    • 2005
  • The water extract of Omijatang(OMJT) has been traditionally used for treatment of abscess and heart palpitation in oriental medicine, However, little is known about the mechanism by which the water extract of OMJT rescues cells from these damages. This study was designed to investigate the protective mechanisms of OMJT in H9c2 cardiomyoblasts on oxidative stress-induced cytotoxicity including $H_2O_2,\;ZnCl_2$, hypoxia, and reoxygenation. Oxidative stress markedly decreased the viability of H9c2 cells. This was characterized with apparent apoptotic features such as chromatin condensation as well as fragmentation of genomic DNA and nuclei. However, OMJT significantly reduced $H_2O_2$-induced cell death and apoptotic characteristics as well as $ZnCl_2$, hypoxialreoxygenation. Taken together, this study suggests that the water extract of OMJT has the protective effects against oxidative injuries.

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Analysis of Gene Expression in 4,4'-Methylenedianiline-induced Acute Hepatotoxicity

  • Oh, Jung-Hwa;Yoon, Hea-Jin;Lim, Jung-Sun;Park, Han-Jin;Cho, Jae-Woo;Kwon, Myung-Sang;Yoon, Seok-Joo
    • Toxicological Research
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    • v.25 no.2
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    • pp.85-92
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    • 2009
  • 4,4'-Methylenedianiline (MDA) is an aromatic amine that is widely used in the industrial synthetic process. Genotoxic MDA forms DNA adducts in the liver and is known to induce liver damage in human and rats. To elucidate the molecular mechanisms associated with MDA-induced hepatotoxicity, we have identified genes differentially expressed by microarray approach. BALB/c male mice were treated once daily with MDA (20 mg/kg) up to 7 days via intraperitoneal injection (i.p.) and hepatic damages were revealed by histopathological observation and elevation of serum marker enzymes such as AST, ALT, ALP, cholesterol, DBIL, and TBIL. Microarray analysis showed that 952 genes were differentially expressed in the liver of MDA-treated mice and their biological functions and canonical pathways were further analyzed using Ingenuity Pathways Analysis (IPA). Toxicological functional analysis showed that genes related to hepatotoxicity such hyperplasia/hyperproliferation (Timp1), necrosis/cell death (Cd14, Mt1f, Timp1, and Pmaip1), hemorrhaging (Mt1f), cholestasis (Akr1c3, Hpx, and Slc10a2), and inflammation (Cd14 and Hpx) were differentially expressed in MDA-treated group. This gene expression profiling should be useful for elucidating the genetic events associated with aromatic amine-induced hepatotoxicity and for discovering the potential biomarkers for hepatotoxicity.

2-Chloroethylethyl Sulfide Induces Apoptosis and Necrosis in Thymocytes

  • Hur, Gyeung-Haeng;Kim, Yun-Bae;Shin, Sung-Ho
    • BMB Reports
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    • v.31 no.2
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    • pp.183-188
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    • 1998
  • 2-chloroethylethyl sulfide (CEES) is an alkylating agent that readily reacts with a wide variety of biological molecules causing metabolic abnormality. The mechanism of cell death during CEES injury is poorly understood. We have examined the effect of exposure of thymocytes with various concentrations of CEES to determine the pattern of cell death in thymocytes injury induced by CEES. In the present study, we show that two patterns of cell death occurred by either one of two mechanisms: apoptosis and necrosis. Exposure to low level of CEES (100 ${\mu}M$) for 5 h caused an induction of apoptosis on thymocytes, as identified by the following criteria: DNA fragmentation visualized by the characteristic "ladder" pattern was observed upon agarose gel electrophoresis and morphological features were revealed by microscopical observations. In contrast, exposure to high levels of CEES (500 ${\mu}M$) induce necrotic features such as cell lysis. Thus, depending on the concentrations, CEES can result in either apoptotic or necrotic cell damage. Our findings suggest that thymocytes which are not killed directly, but merely injured by low levels of CEES, are able to activate an internally-programmed cell death mechanism, whereas thymocytes receiving severe damages apparently can not.

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