• Title/Summary/Keyword: DNA comet assay

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Oxidative Stress of Mouse Fed Irradiated Diet Containing High Unsaturated Fatty Acid (방사선 조사된 고불포화지방산 식이를 섭취한 Mouse의 산화적 스트레스)

  • Seo, Dae-Young;Park, Sun-Young;Kang, Myoung-Hee;Suh, Kwang-Sun;Ly, Sun-Yung
    • Journal of Nutrition and Health
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    • v.39 no.7
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    • pp.599-609
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    • 2006
  • In this study, we investigated the in vivo effect by intake of the irradiated foods such as mackerel and sesame seed which are high in unsaturated fatty acid through TBARS (thiobarbituric acid reactive subtance) and the tissue pathological and genotoxicological test. Thirty two ICR mice are divided into four groups, one non-irradiated (control) group and three irradiated (5, 10, 20 kGy, respectively) groups. Sesame seed and pulverized mackerel in modified AIN93M diet were mixed together then divided into four identical parts. Three parts of them were irradiated by doses of 5, 10, and 20 kGy. These experimental diet were fed to each group for 4, 8 and] 6 weeks. The results of the study were as follows: No significant differences in weight gain were found in each group. Peroxide value of the irradiated diet was higher than that of the non-radiated one and also increased according to the storage period. TBA values in plasma, liver, kidney and Peyer's patch were not significantly different among 4 groups. DNA% in tail, tail length (TL) and tail moment (TM) values of the blood lymphocyte in 4, 8 and 16 week groups and the liver in 16 week group were much measured over the control DNA % in tail of kidney of 8 week group was significantly than the control and TL and TM of 8 week and TM of 16 week groups showed a tendency of higher values. By Peyer's DNA % in tail of 8 week group, DNA % in tail and TM of 16 week groups increased significantly over the control, Ultrastructural examination shows myeline figures and swollen mitochondria in parietal cells and intestinal epithelial cells of 8 and 16 weeks groups. After this study, we need further investigations on the safety of highly consumed foods which contain high contents of unsaturated fatty acid, largely imported and which are possible to be irradiated.

Antioxidant and Antigenotoxic Activities of Extracts from Anglerfish (아귀 추출물의 항산화 및 항유전독성 활성)

  • Lee, Suck-Hee;Shin, Jin-Hwa;Koo, Myoung-O;Jung, Eun-Sil;Jeon, Geong-Im;Park, Eun-Ju;Park, Hae-Ryong;Lee, Seung-Cheol
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.10
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    • pp.1229-1234
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    • 2007
  • Antioxidant activities of extracts from anglerfish (Lophiiu Zitulon) were evaluated. Each part of fresh (skin, flesh, stomach, and liver) or dried (skin and flesh) anglerfish was extracted by four different solvents (methanol, ethanol, acetone, and distilled water). Antioxidant activities of the extracts were determined by radical scavenging activity (RSA) and reducing power (RP). Relatively higher RSA and RP were found in methanol and water extract of fresh anglerfish liver. Antigenotoxic effect of the extracts, which was measured by Comet assay, was shown in most of the extracts except methanol, acetone and distilled water extracts of fresh stomach sample. These results indicated that antioxidant and antigenotoxic properties of extracts from angler fish were variable depending on parts, solvent, and/or physicochemical state. The appropriate extraction process could provide some valuable bioactive materials from anglerfish.

Stability of Functional Properties and Chemical Components of Gamma-Irradiated Astragali Radix (감마선 조사 황기의 기능성 및 화학적 성분에 대한 안정성 평가)

  • Jeong, Ill-Yun
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.2
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    • pp.255-260
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    • 2005
  • Stability of functional properties and chemical components of gamma-irradiated Astragali Radix (AR) were investigated to study the application of radiation technology for preservation and hygiene of medicinal herbs. Thus, ethanolic extracts and ethyl acetate fractions were prepared from gamma-irradiated and gamma-nonirradiated AR, respectively. Functional properties were estimated by comparing antioxidant activity (DPPH radical scavenging activity, lipid peroxidation inhibitory activity) and protective effect against oxidative DNA damage in human lymphocytes. Chemical components were compared by HPLC analysis, comparing with calycosin ($_{t}$ R= 15.111 min), which was isolated from ethyl acetate fraction as a standard material. Their antioxidant activities and protective effects against oxidative DNA damage showed no significant difference at the concentrations tested. HPLC analysis also showed almost same pattem. These results suggest that the functional properties and chemical components of AR are not affected largely by gamma irradiation, and radiation technology can be applied to preservation and hygiene of medicinal herbs.s.

Antioxidant and Antigenotoxic Effects of Sansuyu Fruit (Corni fructus) Extracted with Water at Different Temperatures (추출 온도에 따른 산수유의 항산화 활성 및 항유전독성 효과 비교)

  • Lee, Min-Hee;Kim, Jung-Mi;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.2
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    • pp.149-155
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    • 2011
  • The objective of this study was to evaluate the antioxidant and antigenotoxic activities of sansuyu fruit (Corni fructus, CF) at temperatures of $25^{\circ}C$, $50^{\circ}C$, and $90^{\circ}C$ using a water extraction method. Total phenolic content (TPC), DPPH radical-scavenging activity (RSA), and superoxide dismutase (SOD)-like activity, and ORAC (Oxygen radical absorbance capacity) values were determined. Also the antigenotoxicity of CF was determined by measuring inhibitory effects of $H_2O_2$ induced DNA damage in human leukocytes using the comet assay. The TPC in the CF extracts was 4.2, 4.6, and 5.5 g/100 g GAE in $25^{\circ}C$, $50^{\circ}C$, and $90^{\circ}C$, respectively. The DPPH RSA of the CF extracts increased in a dose-dependent manner over the range of $50\sim1000\;{\mu}g$/mL in all temperatures and the $SC_{50}$ of DPPH RSA of the CF extracts were not significantly different at different extraction temperatures. The $SC_{50}$ of SOD-like was the highest in CF extracted at $25^{\circ}C$ (1.1 mg/mL) followed by $90^{\circ}C$ (1.2 mg/mL) and $50^{\circ}C$ (1.3 mg/mL). The ORAC values of the CF extracts were not significantly different in low concentration ($10\;{\mu}M$/mL) and was in order of $25^{\circ}C$ ($5.7\;{\mu}M$ TE)< $90^{\circ}C$ ($6.2\;{\mu}M$ TE)< $50^{\circ}C$ ($8.5\;{\mu}M$ TE) in high concentration ($50\;{\mu}M$/mL). $200\;{\mu}M$ $H_2O_2$ induced DNA damages in human leukocytes were significantly reduced by the pretreatment with the CF extracts. These results suggest that sansuyu fruit (Corni fructus) can be used as a natural source for antioxidant activities and as antigenotoxic agents regardless of the water extraction temperature.

Antihepatotoxic and Antigenotoxic Effects of Herb Tea Composed of Chrysanthemum morifolium Ramat. (국화차를 포함하는 허브차의 CCl4로 유도된 간세포손상 보호 및 항유전독성 효과)

  • Lee, Hyun-Jung;Hwang, Young-Il;Park, Eun-Ju;Choi, Sun-Uk
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.1
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    • pp.78-83
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    • 2011
  • The flower of Chrysanthemum morifolium Ramat. with antioxidant, anticancer, and anti-inflammatory functions has been a widely used traditional herb as a healthy beverage and medicine. The aim of the present study was to investigate a herb tea consisting of C. morifolium Ramat., Corni fructus and Schizandra chinensis Baillon for its hepatoprotective activity against $CCl_4$-induced toxicity in freshly isolated rat hepatocytes and antigenotoxic effect against oxidative stress induced DNA damage in human leukocytes. Three different compositions of the herb tea (Mix I, II, and III) were prepared by extracting with water at $90^{\circ}C$. Freshly isolated rat hepatocytes were exposed to $CCl_4$ along with/without various concentrations of each tea. Protection of rat primary cells against $CCl_4$-induced damage was determined by the MTT assay. The significant antihepatotoxic effect of the tea was shown in Mix I and II. The increased transaminase (AST and/or ALT) release in media of $CCl_4$ treated hepatocytes was significantly lowered by all the teas tested. The effect of the tea on DNA damage in human leukocytes was evaluated by Comet assay. All teas showed a protective effect against $H_2O_2$-induced DNA damage. From these results, it is assumed that herb tea based on C. morifolium Ramat., Corni fructus and Schizandra chinensis Baillon exerted antihepatotoxic and antigenotoxic effects.

A Study for Bioassay on the HBr Combustion Toxity (생물학적 분석을 통한 HBr의 연소 독성에 관한 연구)

  • Cho, Nam-Wook;Shin, Hyun-Joon;Lee, In-Ku;Oh, Eun-Ha
    • Journal of the Korean Applied Science and Technology
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    • v.29 no.4
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    • pp.545-551
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    • 2012
  • Due to the use of polymeric materials in construction materials, the fire combustion gases that occur in the fire are various. The one of combustion gases, HBr is measured to evaluate the toxicity of the combustion gases in the FTP Code Part 2, Standard NES 713 and Standard BS 6853. According to the MSDS, Inhalation of HBr gas especially cause burn, respiratory dysfunction, headache, etc. The people who are exposed to 50ppm of HBr gas, very irritant gas may also frequently result in both immediate death and post-exposure deaths due to pulmonary complications. In this paper, we conduct a research on the combustion toxicity of HBr gas hazardous test which is motility measurement of the mice exposed to the HBr standard gas comparing the biological analysis result.

Early Activation of Apoptosis and Caspase-independent Cell Death Plays an Important Role in Mediating the Cytotoxic and Genotoxic Effects of WP 631 in Ovarian Cancer Cells

  • Gajek, Arkadiusz;Denel-Bobrowska, Marta;Rogalska, Aneta;Bukowska, Barbara;Maszewski, Janusz;Marczak, Agnieszka
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.18
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    • pp.8503-8512
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    • 2016
  • The purpose of this study was to provide a detailed explanation of the mechanism of bisanthracycline, WP 631 in comparison to doxorubicin (DOX), a first generation anthracycline, currently the most widely used pharmaceutical in clinical oncology. Experiments were performed in SKOV-3 ovarian cancer cells which are otherwise resistant to standard drugs such as cis-platinum and adriamycin. As attention was focused on the ability of WP 631 to induce apoptosis, this was examined using a double staining method with Annexin V and propidium iodide probes, with measurement of the level of intracellular calcium ions and cytosolic cytochrome c. The western blotting technique was performed to confirm PARP cleavage. We also investigated the involvement of caspase activation and DNA degradation (comet assay and immunocytochemical detection of phosphorylated H2AX histones) in the development of apoptotic events. WP 631 demonstrated significantly higher effectiveness as a pro-apoptotic drug than DOX. This was evident in the higher levels of markers of apoptosis, such as the externalization of phosphatidylserine and the elevated level of cytochrome c. An extension of incubation time led to an increase in intracellular calcium levels after treatment with DOX. Lower changes in the calcium content were associated with the influence of WP 631. DOX led to the activation of all tested caspases, 8, 9 and 3, whereas WP 631 only induced an increase in caspase 8 activity after 24h of treatment and consequently led to the cleavage of PARP. The lack of active caspase 3 had no outcome on the single and double-stranded DNA breaks. The obtained results show that WP 631 was considerably more genotoxic towards the investigated cell line than DOX. This effect was especially visible after longer times of incubation. The above detailed studies indicate that WP 631 generates early apoptosis and cell death independent of caspase-3, detected at relatively late time points. The observed differences in the mechanisms of the action of WP631 and DOX suggest that this bisanthracycline can be an effective alternative in ovarian cancer treatment.

The Physicochemical Stabilities and Biological Activities of Pigment Extract from Marine Bacterium Pseudoalteromonas psicida TA20 (해양세균 Pseudoa/teromonas psicida TA20의 색소 추출물의 물리화학적 안정성과 생리활성)

  • Park, Jin-Sook;Jeong, Dong-Woo;Kang, Myung-Hee
    • Journal of Life Science
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    • v.19 no.8
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    • pp.1132-1138
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    • 2009
  • We investigated the physicochemical stabilities and biological activities of ethanol- extracted pigment from marine bacterium Pseudoalteromonas psicida TA20. This bacterial pigment was very stable at a pH range of between 4.0 and 8.0 at a temperature below $40^{\circ}C$. In the presence of light, the pigment was also very stable, showing more than 90 percent remaining absorbance during 14 days at $25^{\circ}C$. The stability of the pigment, when metal ions were present, showed higher stability in all examined metal ions except for $Al^{3+}$ and $Cu^{2+}$, especially in the presence of $Fe^{2+}$. This pigment showed higher stability than other pigment extracts reported. The pigment has free-radical scavenging (3,495 ${\mu}g/ml$) activity and 44% antioxidant protective effect against DNA damage of human lymphocyte cells at a concentration of 10 ${\mu}g/ml$. The results indicate that the bacterial pigment produced a significant reduction in oxidative DNA damage. The pigment also showed antimicrobial activity against major food poisoning bacteria. Therefore, these results suggest that this bacterial pigment could be used as a natural colorant in the food industry, having the advantages of antioxidant and antimicrobial activities.

The Improvement of Chaga Mushroom (Inonotus Obliquus) Extract Supplementation on the Blood Glucose and Cellular DNA Damage in Streptozotocin-Induced Diabetic Rats (Streptozotocin으로 유발한 당뇨쥐에 있어서 차가버섯(Inonotus Obliquus)의 혈당 및 DNA 손상 개선효과)

  • Park, Yoo-Kyoung;Kim, Jung-Shin;Jeon, Eun-Jae;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.42 no.1
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    • pp.5-13
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    • 2009
  • Mushrooms have become a largely untapped source of powerful new pharmaceutical products that poses anti-inflammatory, and antimutagenic, and antioxidant activities. The antioxidant effects of the mushroom may be partly explained by protecting cellular components against free radical. The aim of this study was to investigate the protective effect of chaga mushroom against diabetes, via the mitigation of oxidative stress and reduction of blood glucose, in streptozotocin-induced diabetic rats. Rats were rendered diabetic by intravenous administration of STZ through tail at a dose of 50 mg/kg. Animals were allocated into four groups with 8 rats each. The control and diabetic control group were fed with standard rat feed. The other diabeic groups, the low chaga extract group and the high chaga extract group were fed ad libitum using 0.5 g/kg and 5 g/kg of chaga mushroom extract, respectively, for 4 weeks. The blood glucose levels in the two chaga extract groups showed a tendency to decrease but did not reach statistical significance after the supplementation. Leukocyte DNA damage, expressed as tail length, was found to be significantly lower in the high chaga extract group than in the diabetic control group (p > 0.05). Plasma level of total radical-trapping antioxidant potential (TRAP) was tend to be higher in the high chaga extract group compared with the diabetic control group. Erythrocyte antioxidant enzyme activities of two groups did not differ. Although we did not obtain beneficial effect on lowering blood glucose levels in the STZ-induced diabetic rats, this results suggest that the chaga mushroom extracts may initially act on protecting endogenous DNA damage in the short-term experiment.

Ethanol Induced Leucocytic and Hepatic DNA Strand Breaks Are Prevented by Styela clava and Styela plicata Supplementation in Male SD Rats (알코올로 인한 흰쥐의 백혈구 및 간 DNA 손상에 미치는 미더덕과 오만둥이 분말의 보충섭취 효과)

  • Kim, Jung-Mi;Park, Hae-Ryoung;Lee, Seung-Cheol;Park, Eun-Ju
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.36 no.10
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    • pp.1271-1278
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    • 2007
  • In this study, the ability of Styela clava or Styela plicata to reduce ethanol-induced hepatotoxicity and hepatic and leucocytic DNA damages was evaluated. Twenty four male SD rats were given 25% ethanol containing water (ad lib, p.o.) and divided into 3 groups; ethanol treated control group (EtOH), ethano1+3% S. clava (EtOH+SC), and ethano1+3% S. plicata (EtOH+SP). After 6 weeks, the supplementation of S. clava reduced the plasma ALT, ALP and LDH activities significantly (p<0.05), while S. plicata induced significant decrease in the plasma LDH activity only. The comet assay was employed to quantify the alcohol-induced DNA damage in rat hepatocytes and leucocytes. A significant protective effect on hepatic and leucocytic DNA damages was observed in S. clava or S. plicata supplemented groups compared to the EtOH control group. The hepatic DNA damage was correlated positively with plasma ALP and LDH activities. These results demonstrated that S. clava or S. plicata supplementation protected alcohol-induced hepatic and leucocytic DNA damage.