• Title/Summary/Keyword: human lymphocyte DNA

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Screening of the Extracts of Herbal Medicines which Stimulate the Hydrolysis of Phosphoinositides in Jurkat T-lymphocyte Cells (Jurkat T 면역세포에서 Phosphoinositides의 가수분해를 증가시키는 약용식물 추출물의 검색)

  • 민도식;이영한;백석환;서판길;류성호
    • Biomolecules & Therapeutics
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    • v.4 no.2
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    • pp.148-153
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    • 1996
  • Activation of the T lymphocytes results in a variety of early biochemical events ultimately leading to cell proliferation and lymphokine production. Stimulation of the signal transduction cascade in T cells through the T cell receptor coincides with activation of the phosphatidylinositol-phospholipase C (PI-PLC) pathway. Therefore, we have established a model system to screen immune-simulator that can increase the hydrolysis of phosphoinositides in human T cell leukemia Jurkat cells. As a result of screening from herbal medicine extract, 4 extracts (O1ibanum, Ephedrae Herba, Real Gar, Saussureae Radix) were found 14 increase the production of inositol phosphates. All the active fraction from the four kinds of extract were fluted in a different retention time on C-18 HPLC and these active fraction also showed difference in cell specificity. And all the active fractions increased DNA synthesis in T cell. Therefore, it is suggested that the active fraction among 4 extracts might contain a compound having different properties one another.

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Protective Effect of Edible Mushrooms (Pleurotus ostreatus, Flammulina velutipes, Lentinula edodes) according to Different Cooking Methods on DNA Damage of Jurkat Cell Line (식용 버섯의 조리방법에 따른 Jurkat 세포주 DNA 손상 보호 효과)

  • Cho, Yun-Jeong;Kim, Kyoung-Hee;Yook, Hong-Sun
    • The Korean Journal of Food And Nutrition
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    • v.28 no.1
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    • pp.34-39
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    • 2015
  • In this study, portective effect on DNA damage several mushrooms (Pleurotus ostreatus, Flammulina velutipes, Lentinula edodes) according to cooking methods was investigated using Comet assay. Three edible mushrooms were cooked by grilling, blanching, pan-frying, or by preparing 'Jeon' (traditional Korean pancake). Cells were incubated in medium with 4 kinds of samples for 48 h ($37^{\circ}C$) were further treated with hydrogen peroxide ($H_2O_2$) for 5 min as an oxidative stimulus. Oxidative damage was evaluated by single-cell gel electrophoresis (Comet assay) and quantified by tail DNA% (TD), tail length (TL), tail moment (TM). Though oxidative DNA damages expressed as TD, TL, TM of 4 cooked samples were higher than raw sample, which means lower protective activities, all samples including raw sample had significantly higher protective effects than the positive control (p<0.05). The protective effect on DNA damage of cooked samples decreased much more when soybean oil added, likely due to the thermal oxidation of oil during cooking. Although heat treatment could degrade protective effect on DNA damage of mushrooms, the cooked mushroom had significant effect on oxidative stress. In conclusion, grilling and blanching were the most advantageous cooking methods to protect oxidative DNA damage induced by $H_2O_2$.

GENE EXPRESSION CHARACTERISTICS OF PUTATIVE PROINFLAMMATORY CYTOKINES AND RECEPTOR MOLECULE CLONING (Putative proinflammatory cytokine유전자의 발현양상과 수용체 분자의 cloing)

  • Oh, Kwi-Ok;Song, Yo-Han;Seo, Young-Seok;Lee, Dong-Whan;Moon, Dae-Hee;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
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    • v.24 no.3
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    • pp.472-482
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    • 1994
  • Cytokines expressed specifically in leukocytes subsets and in activated cells, which are involved in chemotaxis and activation of leukocytes, are recently defined as chemokines. Macrophage inflammatory $protein-1{\alpha}(MIP-1{\alpha})$ and $MIP-1{\beta}$ are members of C-C chemokine subfamily which produces wide immunomodulatory, proinflammatory, and hematopoietic modulatory actions. We have studied their gene expression by using Northern blot analysis in various blood cells such as cytolytic T lymphocyte(CTL), helper T lymphocyte(HTL), macrophage, and B lymphocyte. Resting CTL line CTLL-R8 expressed $MIP-1{\alpha}$ mRNA which was downregulated by ConA stimulation. Both of resting and ConA stimulated HTL line Hut78 and Jurkat did not express $MIP-1{\alpha}$ mRNA. There was detectable $MIP-1{\alpha}$ transcript in HTL hybridoma 2B4.11 which was a little upstimulated by ConA stimulation. B cell line 230, and macrophage cell line RAW264.7 and WR19M.1 showed distinct $MIP-1{\alpha}$ message which were induced after LPS stimulation. Expression pattern of $MIP-1{\beta}$ in all cell lines or cell were almost identical to that of $MIP-1{\alpha}$. Also strategies employed to identify and characterize the biological functions was preceded by receptor cloning to trace the shorcut to the final goal of cytokine research. For the cloning of $MIP-1{\alpha}$ receptor(R), we used synthetic oligonucleotides of transmembrane(T) conserved sequences of already cloned human(h) IL-8-R, and performed reverse transcription-polymerase chain reaction(RT-PCR) amplification using murine(m) macrophage cell line mRNA. Among 5RT-PCR products, we isolated a homologous cDNA with hIL-8-R which were shown to be putative mIL-8-R cDNA.

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Comparative Study on Human Risk by Ionizing Radiation and Pesticide as Biological Information about Environmental Disaster (환경재해에 관한 생물정보로서의 이온화 방사선과 살충제의 인체 위해성 비교 연구)

  • Kim, Jin-Kyu;Hyun, Soung-Hee
    • Journal of Radiation Protection and Research
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    • v.26 no.4
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    • pp.385-392
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    • 2001
  • Environmental risk factors such as ionizing radiations, heavy metals, and pesticides can cause environmental disasters when they exist in excess. The increases in use of ionizing radiation and agricultural pesticide are somewhat related to the possibility of the disaster. The risk of radiation and pesticide was evaluated by means of the single cell gel electrophoresis (SCGE) assay on the human blood lymphocytes. The lymphocytes were irradiated with $0{\sim}2.0Gy$ of $^{60}Co$ gamma ray. Another groups of lymphocytes were exposed to various concentrations of parathion. Significantly increased tail moment, which was a marker of DNA strand breaks in SCGE assay, showed a clear dose- or concentration-response relationship. Parathion of a recommended concentration for agricultural use ($1mg {\ell}^{-1}$ ) has a strong cytotoxic effect on lymphocytes, which is equivalent to damage induced by 0.1 Gy of ${\gamma}$-ray. Furthermore, $2mg{\ell}^{-1}$ of parathion can give rise to DNA damage equivalent to that induced by 0.25 Gy at which the radiation-induced damage can start to develop into clinical symptoms. The comparative results of this study can provide an experimental basis and biological information for the prevention of environmental disaster.

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Protective Effect of Flavonoids on Lymphocyte DNA Damage Using Comet Assay (Comet Assay를 이용한 Flavonoids와 항산화 비타민의 인체임파구 세포 DNA 손상 보호 효과)

  • 박유경;전은재;강명희
    • Journal of Nutrition and Health
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    • v.36 no.2
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    • pp.125-132
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    • 2003
  • The present study was attempted to investigate and compare the antioxidant potency of several well-know flavonoids, antioxidant vitamin and commercially available popular beverages. The antioxidant potency was assessed by the effect on reducing oxidative DNA damage of human lymphocytes. Cellular oxidative DNA damage was measured by SCGE (single-cell gel electrophoresis), also known as comet assay. Lymphocytes were pre-treated for 30 minutes with wide ranges of doses of apigenin, kaempferol, luteolin, myricetin, rutin, quercetin, $\alpha$-tocopherol (10,25,50,100,200,500,1000 $\mu$M) ,green tea extract or grape juice (10,50,100,250,500,1000 $\mu$g/mL) followed by a $H_2O$$_2$(100 $\mu$M) treatment for 5 min as an oxidative stimulus. The physiological function of each antioxidant substance on oxidative DNA damage was analyzed as tail moment (tail length $\times$ percentage migrated DNA in tail) and expressed as relative DNA damage score after adjusting by the level of control treatment. Cells treated with $H_2O$$_2$alone (positive control) had an extensive DNA damage compared with cells treated with phosphate buffered saline (PBS, negative control) or pre-treated with all the tested samples. Of all the six flavonoids, quercetin was the most potent antioxidant showing the lowest $ED_{50}$/ of 8.5 $\mu$g/mL (concentration to produce 50% protection of relative DNA damage). The antoxidant potency of individual flavonoids were ranked as follows in a decreasing order; luteolin (18.4 $\mu$g/mL), myricetin (19.0 $\mu$g/mL) , rutin (22.2 $\mu$g/mL) , apigenin (24,3 $\mu$g/mL) , kaempferol (25.5 $\mu$g/mL). The protective effect of $\alpha$-tocopherol was substantially lower (highest $ED_{50}$value of 55.0 $\mu$g/mL) than all the other flavonoids, while the protective effect was highest in green tea and grape juice with low ED5O value of 7.6 and 5.3, respectively. These results suggest that flavonoids, especially quercetin, and natural compounds from food product, green tea and grape juice, produced powerful anti-oxidative activities, even stronger than $\alpha$-tocopherol. Taken together, supplementation of antioxidants to lymphocytes followed by oxidative stimulus inhibited damage to cellular DNA, supporting a protective effect against oxidative damage induced by reactive oxygen species.

Association of the -2518 A/G Polymorphism of MCP-1 with Breast Cancer in Punjab, North-West India

  • Sambyal, Vasudha;Guleria, Kamlesh;Kapahi, Ruhi;Manjari, Mridu;Sudan, Meena;Uppal, Manjit Singh;Singh, Neeti Rajan
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7243-7248
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    • 2015
  • Background: Monocyte chemoattractant protein-1 (MCP-1) is a major chemokine thought to be responsible for monocyte and T-lymphocyte recruitment in acute inflammatory conditions and recruitment of macrophages in tumors. It is also implicated in cardiovascular disease, rheumatoid arthritis and chronic obstructive pulmonary disease. The aim of the present study was to investigate the correlation between MCP-1 -2518 A/G polymorphism and breast cancer risk in patients from Amritsar city of Punjab state in North-West India. Materials and Methods: We screened DNA samples of 200 sporadic breast cancer patients and 200 age and gender matched unrelated healthy individuals for MCP-1 -2518 A/G polymorphism using the PCR-RFLP method. Results: A significantly increased frequency of the GG genotype was observed in patients as compared to controls. Individuals carrying the MCP1 -2518GG genotype had a two fold risk for breast cancer (OR=2.06, 95%CI, 1.06-3.98; p=0.03). Genetic models analysis revealed a significant association between MCP-1 -2518 A/G polymorphism and cancer risk in homozygous co-dominant (OR=2.06, 95%CI, 1.06-3.98; p=0.03) and recessive (OR=1.97, 95%CI, 1.05-3.70; p=0.03) models. Conclusions: We conclude that the GG genotype of the MCP-1-2518 A/G polymorphism is associated with increased risk to breast cancer in Punjab, North-West India.

Protective effect of Korean diet food groups on lymphocyte DNA damage and contribution of each food group to total dietary antioxidant capacity (TDAC) (한식 식품군의 in vitro 총 항산화능 (TDAC)과 ex vivo DNA 손상 보호효과와의 관련성)

  • Lee, Min Young;Han, Jeong-Hwa;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.49 no.5
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    • pp.277-287
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    • 2016
  • Purpose: This study was performed to compare total phenolic contents, in vitro antioxidant capacity, and reduction effect of Korean food groups on ex vivo DNA damage in human cells and analyze correlations between each indicator. Methods: Vegetable foods in the Korean diet based the results of the KNHANES V-2 (2011) were classified into 10 food groups: cereals, fruits, vegetables, nuts, kimchi, seaweeds, potatoes, mushrooms, legumes, and oils. Eighty-four foods constituted more than 1% of the total intake in each food group and finally designated as vegetable foods in the Korean diet. Total phenolic content of each food group was measured. Further, in vitro antioxidant capacity was measured based on DPPH radical scavenging assay, TEAC assay, and $ORAC_{ROO{\cdot}}$ assay. Ex vivo DNA damage in human lymphocytes was assessed using comet assay. Results: Total phenolic contents of food groups of the Korean diet increased in the order of mushrooms, fruits, vegetables, seaweeds, and kimchi. Meanwhile, antioxidant rankings of food groups as mean values from the three in vitro test methods increased in the order of mushrooms, seaweeds, vegetables, kimchi, and fruits. Protection against ex vivo DNA damage in human lymphocytes was highest in mushrooms, followed by vegetables, fruits, seaweeds, and kimchi. The rankings of the food groups for total phenolic content, in vitro DAC, and ex vivo DNA protection activity were similar, and correlations between each indicator were significantly high. Conclusion: Mushrooms, fruits, vegetables, and seaweeds among the tested food groups in the Korean diet showed high total phenolic contents, in vitro antioxidant capacities, and protection against DNA damage. Correlations between each indicator in terms of total phenolic content, in vitro antioxidant capacity, and ex vivo DNA protection between each food group were found to be particularly high.

Effects of lymphocyte DNA damage levels in Korean plant food groups and Korean diet regarding to glutathione S-transferase M1 and T1 polymorphisms (건강한 성인의 glutathione S-transferase M1과 T1 유전자 다형성에 따른 한식에서의 식물성 식품군과 한식의 DNA 손상 감소 효과)

  • Kim, Hyun-A;Lee, Min-Young;Kang, Myung-Hee
    • Journal of Nutrition and Health
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    • v.50 no.1
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    • pp.10-24
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    • 2017
  • Purpose: GST (glutathione S-transferase) M1 and T1 gene polymorphisms are known to affect antioxidant levels. This study was carried out to evaluate genetic susceptibility by measuring the effect of DNA damage reduction in the Korean diet by vegetable food according to GST gene polymorphisms using the ex vivo method with human lymphocytes. Methods: Vegetable foods in the Korean diet based the results of the KNHANES V-2 (2011) were classified into 10 food groups. A total of 84 foods, which constituted more than 1% of the total intake in each food group, were finally designated as a vegetable food in the Korean diet. The Korean diet applied in this study is the standard one-week meals for Koreans (2,000 Kcal/day) suggested by the 2010 Dietary Reference Intakes for Koreans. Ex vivo DNA damage in human lymphocytes was assessed using comet assay. Results: In the Korean food group, the DNA damage protective effect of GSTM1 and GSTT1 was found to be greater in mutant type and wild-type, respectively. and the DNA damage protective effect according to the combined genotype of GSTM1 and GSTT1 was different depending on the food group. On the other hand, in Korean Diet, the DNA damage protective effect appeared to be larger in GSTM1 wild-type than in mutant type and was found to not be affected by GSTT1 genotype. Conclusion: These results can be used as basic data to demonstrate the superiority of the antioxidant function of Korean dietary patterns and food groups. Furthermore, it may be a starting point to begin research on customized antioxidant nutrition according to individual genes.

Comparison of antioxidant activity and prevention of lymphocyte DNA damage by fruit and vegetable juices marketed in Korea (시판 천연 과일주스와 채소주스의 항산화능과 임파구 DNA 손상 방지 효능 비교)

  • Cho, Miran;Lee, Hye-Jin;Kang, Myung-Hee;Min, Hyesun
    • Journal of Nutrition and Health
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    • v.50 no.1
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    • pp.1-9
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    • 2017
  • Purpose: Fruit and vegetable juices are known to be rich sources of antioxidants, which have beneficial effects on diseases caused by oxidative stress. The purpose of this study was to directly compare the antioxidant activities of fruit and vegetable juices marketed in Korea. Methods: We analyzed four fruit juices, two vegetable juices, two yellow-green juices, and six mixed vegetable juices. Antioxidant activities were analyzed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) test, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonate) (ABTS) test, and oxygen radical absorbance capacity (ORAC) assay. Protective effects against DNA damage were determined using an ex vivo comet assay with human lymphocytes. Results: DPPH radical scavenging activities were in the following order: blueberry juice > mixed vegetable C juice > kale juice > mixed vegetable P juice > grape juice. ABTS radical scavenging activities were in the following order: blueberry juice > mixed vegetable C juice > grape juice > mixed vegetable P juice > kale juice. Peroxyl radical scavenging activities as assessed by ORAC assay were in the following order: blueberry juice > kale juice > mixed vegetable C juice > grape juice. Grape or blueberry juice showed strong abilities to prevent DNA damage in lymphocytes, and the difference between them was not significant according to the GSTM1/GSTT1 genotype. Conclusion: Antioxidant activities of fruit and vegetable juices and ex vivo DNA protective activity increased in the order of blueberry juice, grape juice, and kale juice, although the rankings were slightly different. Therefore, these juices rich in polyphenols and flavonoids deserve more attention for their high antioxidant capacity.

Genotoxicity of low-dose Glyphosate by Sister Chromatid Exchange (자매염색분체교환을 통한 글라이포세이트 유전독성)

  • Lee, Sang Hoon;Kim, Sung Jin;Choi, Woo Ik;Jin, Sang Chan;Choi, In Jang;Lee, Jae Ho
    • Journal of The Korean Society of Clinical Toxicology
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    • v.12 no.1
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    • pp.8-13
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    • 2014
  • Purpose: Glyphosate (N-phosphonomethyl glycine) is widely used as an herbicide for weed control in rural areas. It is also readily available for suicide attempts. Glyphosate has high toxicity and negatively affects the human body. The aim of this investigation was to study the genotoxicity of a low-concentration of glyphosate through sister chromatid exchange (SCE) in human blood lymphocytes in vitro. Methods: Primary lymphocyte cultures were obtained from blood samples of 11 males and seven females who had been exposed to glyphosate (0, 100, 200, and 300 ng/mL). The frequency of SCEs was examined and statistical analysis was performed. Results: All doses of glyphosate induced a significant dose-dependent increase in SCE frequency compared with the control group (P<0.001). In particular, the SCE frequency for exposure to low-dose glyphosate was significantly higher in females than in males. Conclusion: According to the result of this study, even a low-dose of glyphosate may damage DNA and females are more vulnerable to glyphosate.

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