• Title/Summary/Keyword: Biological damage

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Effects of a Phosphomimetic Mutant of RAP80 on Linear Polyubiquitin Binding Probed by Calorimetric Analysis

  • Thach, Thanh Trung;Jee, Jun-Goo;Lee, Sang-Ho
    • Bulletin of the Korean Chemical Society
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    • v.33 no.4
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    • pp.1285-1289
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    • 2012
  • RAP80 plays a key role in DNA damage responses by recognizing K63-linked polyubiquitin moieties through its two ubiquitin-interacting motif (UIM) domains. The linker between the two UIMs possesses a phosphorylation site, but the relationship between phosphorylation and polyubiquitin recognition remains elusive. We investigated the interaction between a phosphorylation-mimic RAP80 mutant S101E and linear polyubiquitins, structurally equivalent to the K63-linked ones, using isothermal titration calorimetry (ITC). ITC analysis revealed differential binding affinities for linear tetraubiquitin by otherwise equivalent UIMs in S101E. Mutational analysis supported such differential polyubiquitin recognition by S101E. Our results suggest a potential crosstalk between polyubiquitin recognition and phosphorylation in RAP80.

A $G_{4}$ Sequence within PHR1 Promoter Acts as a Gate for Cross-Talks between Damage-Signaling Pathway and Multi-Stress Response

  • Jang, Yeun-Kyu;Kim, Eun-Mi;Park, Sang-Dai
    • Animal cells and systems
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    • v.6 no.3
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    • pp.271-275
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    • 2002
  • Rph1 and Gisl are damage-responsive repressors involved in PHR1 expression. They have two $C_{2}$H/ sub 2/ zinc finger motifs as putative DNA binding domains and N-terminal conserved domain with unknown function. They are also found in the human retinoblastoma binding protein 2 and the mouse jumonji- encoded protein. The repressors are able to bind to A $G_{4}$ sequence within a 39-bp sequence called upstream repressing sequence of PHR1 promoter (UR $S_{PHR1}$) responsible for the damage-response of PHR1. We report here that Rph1 is predominantly localized in the nucleus as examined by fluorescence microscopic analysis with GFP-Rph1 fusion protein. On the basis of the fact that the A $G_{4}$ sequence that is recognized by Rph1 and Gisl is also recognized by Msn2 and Msn4 in a process of stress response, we a1so tried to examine the in vivo function of A $G_{4}$ and the role of Msn2 and Msn4 in PHR1 expression. Our results demonstrate that Msn2 and Msn4 are actually required for the basal transcription of PHR1 expression but not for its damage induction. When A $G_{4}$ sequence was inserted into the minimal promoter of the cyc1-LacZ reporter, the increased LacZ expression was observed indicating its involvement in transcriptional activation. The data suggest that the A $G_{4}$ is primarily required for basal transcriptional activation of PHR1 or CYC1 promoter through the possible involvement of Msn2 and Msn4. However, since the A $G_{4}$ is also involved in the repression of PHR1 via Rphl and Gisl, it is proposed that A $G_{4}$ functions as either URS or upstream activating sequence (UAS) depending on the promoter context.t.

The visible injury and physiological responses of three varieties of hot peppers to ozone

  • Kim, Bo-Sun;Yun, Sung-Chul
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.93.1-93
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    • 2003
  • A growth chamber fumigation was conducted to evaluate the ozone (O3) on the physiology of three hot pepper, Capsicum annuum L. cultivars, 'dabotab', 'buchon' and 'pochungchun'. Thirty-day old plants were exposed to O3 of 120 nl 1-1 in the chambers for 8 h d-1 for 3 days. Foliar damage due to O3 was different from the varieties, 'dabotab'was most sensitive to O3, 'pochungchun' was medium, and 'buchon' was resistant. Ozone symptom on the leaves was bifacial necorsis. Photosynthesis and stomatal conductance were decreased due to O3 treatment, but they were not much different from the variety. Decreases of net photosynthesis by O3 were 56%, 40% and 35% on 'dabotab', 'buchon' and 'pochungchun', respectively Decreases of stomatal conductance by O3 were 66%, 63%, and 50% on each varieties. Ozone closed the stomata and decrease net photosynthesis on hot peppers regardless of the variety. Light curves on the three varieties were showing similar patterns that O3 damage on net photosynthesis were started at the low levels of light with or without the visible injury, Assimilation-internal CO2 concentration curves of the three cultivars were not different due to the treatment. It means there was not significant biochemical damage Inside the leaves by O3. In conclusion, ozone closed the stomata and damaged light capturing system of the pepper leaves with or without the visible damage. Although visible damage of the leaves could be a good indicator of O3 resistance, the ecophysiological change by O3 were not proportional to the amout of visible injuries

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Protective Effect of Electrolyzed Reduced Water on the Paraquat-induced Oxidative Damage of Human Lymphocyte DNA (Paraquat에 의한 사람 임파구 DNA 손상에 대한 환원전리수의 보호효과)

  • Park, Eun-Ju;Ryoo, Kun-Kul;Lee, Yoon-Bae;Lee, Jong-Kwon;Lee, Mi-Young
    • Applied Biological Chemistry
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    • v.48 no.2
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    • pp.155-160
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    • 2005
  • Electrolyzed reduced water (ERW), showing extremely negative oxidation-reduction potential, was used to investigate the effects of paraquat-induced damages on DNA from human lymphocyte. The effect of ERW on paraquat-induced oxidative DNA damage in human lymphocytes was evaluated by Comet assay (single-cell gel electrophoresis) quantified as percentage fluorescence in tail. Comet assay has been used widely to assess the level of the DNA damage in individual cells. Lymphocytes were oxidatively challenged with various concentrations of paraquat for 30 min at $37^{\circ}C$, and were then treated with electrolyzed reduced water for 30 min. The oxidative DNA damage by paraquat, as indicated by the fluorescent tail in DNA, increased in a dose-dependent manner. However, oxidative damage of the DNA was almost completely prevented upon treatment with electrolyzed reduced water.

Inhibitory Effect of Capsaicin against Carcinogen-induced Oxidative Damage in Rats

  • Yu, Ri-Na;Park, Min-Ah;Kawada, Teruo;Kim, Byung-Sam;Han, In-Seob;Yoo, Hoon
    • Preventive Nutrition and Food Science
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    • v.7 no.1
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    • pp.67-71
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    • 2002
  • Capsaicin (trans-8-methyl-N-vanillyl-6-nonenarnide), a major pungent component of hot pepper, is known to exert antioxidative properties. In this study, we investigated the protective effects of capsaicin against chemical carcinogen-induced oxidative damage in rats. Male Sprague Dawley rats weighting 230~250 g were treated with chemical carcinogens such as 2-nitropropane (2NP) or n-methyl-N'-nitro-N-nitrosoguanidine (MNNG) after (or before) the administration of capsaicin at doses of 0.5, 1,5 mg/kg. The level of lipid peroxidation in rat liver was estimated by measuring the amounts of thiobarbituric acid reactive substances. The degree of oxidative DNA damage was evacuated by measuring a DNA adduct, 8-hydroxydeoxyguanosine (8-OHdG), in urine. Antioxidative activities of capsaicin and its metabolites in vitro were determined by the measurement of DPPH (1,1-diphenyl-2-picrylhydrazyl), a radical quencher. Significant inhibition of 2-NP induced lipid peroxidation was observed in the liver of the rat when treated with capsaicin. MNNG-induced urinary excretion of 8-OHdG was decreased by capsaicin treatment. Capsaicin and its metabolites inhibited net only the formation of free radicals, but also lipid peroxidation in vitro. Our results show that capsaicin may function as a free radical scavenger against chemical carcinogen-induced oxidative cellular damage in vivo. The observed antioxidative activities of capsaicin may play an important role in the process of chemoprevention.

The linker connecting the tandem ubiquitin binding domains of RAP80 is critical for lysine 63-linked polyubiquitin-dependent binding activity

  • Cho, Hyun-Jung;Lee, Sang-Ho;Kim, Hong-Tae
    • BMB Reports
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    • v.42 no.11
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    • pp.764-768
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    • 2009
  • The tandem ubiquitin-interacting motif (UIM) domain located at the N-terminus of Receptor Associated Protein 80 (RAP80) plays a crucial role in ionizing radiation (IR)-induced DNA damage response. RAP80 translocates to sites of IR-induced DNA damage through interaction of its UIM domain with ubiquitinated H2A and Lys63-linked polyubiquitin chains. The exact mechanism, however, through which RAP80 associates with Lys63-linked polyubiquitin chains is not clear. Here, we show by in vitro GST-pull down assays that modifying the linker region between the tandem ubiquitin binding domains of RAP80 changes the binding affinity for Lys63-linked polyubiquitin chains and affects translocation to sites of DNA breaks. Based on these findings, we suggest that the length of the linker region between the tandem ubiquitin binding domains of RAP80 may be a key factor in the binding of RAP80 with Lys63-linked polyubiquitin chains as well as in the translocation of RAP80 to DNA break sites.

Effect of a Static Magnetic Field on Susceptibility to Ethanol-Induced Hepatic Dysfunction in Rats

  • Park, Kap-Joo;Kim, Eun-Jung;Cho, Myung-Hwan;Lee, Jae-Seok
    • Food Science and Biotechnology
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    • v.15 no.6
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    • pp.833-837
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    • 2006
  • To determine whether alcohol-treated rat liver cells can be protected by a static magnetic field (SMF), we analyzed the blood chemistry and histology of hepatic tissue removed from alcohol-exposed rats that had been exposed to a static magnetic field. The rats were exposed to a 0.3 tesla (3,000 gauss) magnetic field (MF) for 24 hr daily for 5 weeks with appropriate controls. Glutamic pyruvic transaminase activity and the triglyceride levels in animals exposed to the north (N) or south (S) pole of the MF decreased significantly (p<0.01 and p<0.05, respectively) compared with negative control animals with alcohol exposure. A histological examination of hepatic tissue revealed a moderate to severe accumulation of fat vacuoles of various sizes in the cytoplasm of the hepatocytes of animals in the negative control group throughout the study; whereas in groups exposed to the MF poles, fewer fat vacuoles were seen compared with the negative control group. Electron microscopic observations showed that exposure to the N or S pole protected organelles, including the nucleus, from damage during exposure to this toxic agent, as indicated by the fact that the nucleus and the mithochondria virtually retained their shape throughout this study. These results suggest that exposure to a SMF could be an excellent way of protecting against alcohol-induced damage to the rat liver cell.

Proposal of enhanced treatment process based on actual pilot plant for removal of micropharmaceuticals in sewage treatment plants

  • Lee, Shun-hwa;Park, Yun-kyung;Lee, Miran;Lee, Byung-dae
    • Environmental Engineering Research
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    • v.25 no.4
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    • pp.588-596
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    • 2020
  • This study was carried out to increase the treatment efficiency through the improvement of the conventional biological process, and to propose the optimal treatment direction. The optimal treatment conditions were derived based on the results of the spike damage tests in each single process. The removal efficiency of micropharmaceuticals was further increased when an ozone treatment process was added to the biological process compared to the single process. The soil and activated carbon adsorption process was introduced in the post-treatment to remove the micropharmaceutical residues, and the removal efficiency of the pharmaceduticals in the final effluent was more than 85% in spike damage experiment. In particular, the continuous process of biological treatment-ozone-adsorption could ensure the stable treatment of micropharmaceuticals, which had not been efficiently removed in the single process, as it showed more than 80% removal efficiency. Therefore, it is expected that the addition of the ozone oxidation and activated carbon adsorption process to the existing sewage treatment facilities can contribute to the efficient removal of micropharmaceuticals.

An Experimental Study on the Combined Effect of Installation Damage and Creep of Geogrids (지오그리드의 시공시 손상 및 크리프 복합효과에 대한 실험적 연구)

  • Cho, Sam-Deok;Lee, Kwang-Wu;Oh, Se-Yong;Lee, Do-Hee
    • Proceedings of the Korean Geotechical Society Conference
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    • 2005.03a
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    • pp.561-568
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    • 2005
  • The factors affecting the long-term design strength of geogrid can be classified into factors on creep deformation, installation damage, temperature, chemical degradation and biological degradation. Especially, creep deformation and installation damage are considered as main factors to determine the long-term design strength of geogrid. Current practice in the design of reinforced soil is to calculate the long-term design strength of a reinforcement damaged during installation by multiplying the two partial safety factors, $RF_{ID} and RF_{CR}$. This method assumes that there is no synergy effect between installation damage and creep deformation of geogrids. Therefore, this paper describes the results of a series of experimental study, which are carried out to assess the combined effect of installation damage and creep deformation for the long-term design strength of geogrid reinforcement. The results of this study show that the tensile strength reduction factors, RF, considering combined effect between installation damage and creep deformation is less than that calculated by the current design method.

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Antimicrobial efficacy of endophytic Penicillium purpurogenum ED76 against clinical pathogens and its possible mode of action

  • Yenn, Tong Woei;Ibrahim, Darah;Chang, Lee Kok;Ab Rashid, Syarifah;Ring, Leong Chean;Nee, Tan Wen;Noor, Muhamad Izham bin Muhamad
    • Korean Journal of Microbiology
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    • v.53 no.3
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    • pp.193-199
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    • 2017
  • This study was aimed to evaluate the antimicrobial activity of Penicillium purpurogenum ED76 on several clinically important microorganisms. The endophytic fungus P. purpurogenum ED76 was previously isolated from Swietenia macrophylla leaf. The antimicrobial efficacy of P. purpurogenum ED76 dichloromethane extract was determined via disc diffusion and broth microdilution assay. A kill curve study was conducted and the morphology of extract treated bacterial cells were viewed under scanning electron microscope. The dichloromethane extract showed significant inhibitory activity on 4 test bacteria and 2 test yeasts. The minimal inhibitory concentration of the extract ranged from 125 to $1,000{\mu}g/ml$, which indicates the different susceptibility levels of the test microorganisms to the fungal extract. The kill curve study has revealed a concentration-dependent inhibition for all test microorganisms. With the increase of the extract concentration, the microbial growth was significantly reduced. The scanning electron micrograph of dichloromethane extract-treated Staphylococcus aureus cells showed the total damage of the cells. The cell wall invagination of the bacterial cells also indicates the loss of cellular materials and metabolic activity. The gas chromatography mass spectrometry analysis of the extract also showed that the major compound was stigmasterol, which constitutes 45.30% of the total area. The dichloromethane extract of P. purpurogenum ED76 exhibited significant inhibitory activity on several clinically important bacteria and yeasts. The study proposed a possible mode of action that the extract cause significant damage to the morphology of S. aureus cells.