• Title/Summary/Keyword: DNA damaging

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Effects of Vespae Nidus on Peroxynitrite Production and Protein Expression of Proinflammatory Mediators (노봉방(露蜂房)이 t-butylhydroxyperoxide에 의한 Peroxynitrite 생성과 염증성 단백질 발현에 미치는 영향)

  • Jang, Jae-Shik;Jeong, Ji-Cheon;Shin, Hyeon-Cheol
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.6
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    • pp.1499-1505
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    • 2007
  • Peroxynitrite ($ONOO^-$) is a reactive oxidant formed from superoxide anion radical (${\cdot}\;O_2-$) and nitric oxide (NO), which can oxidize cellular components such as essential protein, non-protein thiols, DNA, low-density lipoproteins and membrane phospholipids. ${\cdot}\;O_2-$ and $ONOO^-$ have contributed to the pathogenesis of diseases such as stroke, heart disease, Alzheimer's disease and atherosclerosis. Because of damaging effects of ${\cdot}\;O_2-$ and $ONOO^-$ oxidants, Vespae Nidus, which has been known to strengthen the kidneys to preserve the vital energy. was tested as a potential specific scavenger of those oxidants. In this study, the viability of Vespae Nidus (1, 10, 50 g/ml) to scavenge ${\cdot}\;O_2-$, NO, $ONOO^-$ and so to protect cells against tert-butylhydroxyperoxide (t-BHP) induced cell death was tested. The levels of ${\cdot}\;O_2-$ and $ONOO^-$ were detected by staining with DCFH-DA and DHR 123, respectively. Protein expression levels of COX-2, iNOS and $NF{-\kappa}B$ were assayed by western blot. Vespae Nidus blocked t-BHP-induced cell death in a dose-dependent fashion. Vespae Nidus inhibited t-BHP-induced production of ${\cdot}\;O_2-$, NO and $ONOO^-$ in YPEN cells. The lipid peroxide level was increased and glutathione level was decreased in lipopolysaccharide (LPS)-treated ICR mouse, whereas the ones in the Vespae Nidus-administered group were regulated beneficially. Vespae Nidus inhibited the expression of COX-2, iNOS and NF-κB (p65 and p50) genes in LPS-treated ICR mouse. The present study suggests that Vespae Nidus is a powerful antioxidant and promotes cellular defense activity by scavenging the toxic oxidants such as ${\cdot}\;O_2-$ and $ONOO^-$.

Protective Effects of Ecklonia cava Film on UV-B-induced Photodamages (감태(Ecklonia cava)를 이용한 자외선 차단 필름의 UV-B 조사에 의한 광손상으로부터 보호효과)

  • Lee, Hyo Geun;Won, Yu Sun;Koh, Eun Byeol;Kim, Yoon Ah;Kim, Jeong Eun;Kim, Yoon Jeong;Han, Chae Won;Choi, Min-Woo;Kim, Jae-Il;Jeon, You-Jin
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.6
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    • pp.714-720
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    • 2017
  • Exposure to ultraviolet (UV) radiation is associated with the development of adverse effects in skin. Among the three types of UV rays, UV-B causes the most damaging effects, inducing sunburn and penetrating the outer skin, resulting in DNA mutations and skin cancer. The objective of this study was to formulate a UV-protective film by incorporating Ecklonia cava extracts. Cells covered with the film were exposed to UV-B (50, 80, and $100mJ/cm^2$). To determine the protective effects of the film, we evaluated cell viability, intracellular ROS generation, and apoptosis. We found that all E. cava extracts absorbed UV light and exhibited protective effects against UV-B-induced photodamage. Among the protective films examined in this study, that incorporating an E. cava 70% ethanol extract (70EX) formed the most effective protection against UV-B in HaCaT cells. These findings suggest that the application of film containing E. cava extract could prevent UV-B-induced photodamage, and offer protection against the detrimental effects of UV radiation, thus maintaining physiological condition.

Geographic Genetic Contour of A Leaf Beetle, Chrysolina aurichalcea (Coleoptera: Chysomelidae), on the Basis of Mitochondrial COI Gene and Nuclear ITS2 Sequences

  • Park, Joong-Won;Park, Sun-Young;Wang, Ah-Rha;Kim, Min-Jee;Park, Hae-Chul;Kim, Ik-Soo
    • International Journal of Industrial Entomology and Biomaterials
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    • v.23 no.1
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    • pp.155-166
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    • 2011
  • The leaf beetle, $Chrysolina$ $aurichalcea$ (Coleoptera: Chysomelidae), is a pest damaging plants of Compositae. In order to understand the genetic diversity and geographic variation we sequenced a portion of mitochondrial COI gene (658 bp) and complete nuclear internal transcribed spacer 2 (ITS2) of the species collected from seven Korean localities. A total of 17 haplotypes (CACOI01~CACOI17), with the maximum sequence divergence of 3.04% (20 bp) were obtained from COI gene sequence, whereas 16 sequence types (ITS2CA01~ITS2CA16), with the maximum sequence divergence of 2.013% (9 bp) were obtained from ITS2, indicating substantially larger sequence divergence in COI gene sequence. Phylogenetically, the COI gene provided two haplotype groups with a high nodal support (${\geq}87%$), whereas ITS2 provided only one sequence type group with a high nodal support (${\geq}92%$). The result of COI gene sequence may suggest the presence of historical biogeographic barriers that bolstered genetic subdivision in the species. Different grouping pattern between COI gene and ITS2 sequences were interpreted in terms of recent dispersal, reflected in the ITS2 sequence. Finding of unique haplotypes and sequence types only from Beakryeng-Islet population was interpreted as an intact remnant of ancient polymorphism. As more samples are analyzed using further hyper-variable marker, further fruitful inference on the geographic contour of the species might be available.

Therapeutic strategies to manage chronic wounds by using biofilm dispersal mechanisms (생물막 분산기작을 이용한 만성창상의 치료전략)

  • Kim, Jaisoo;Kim, Min-Ho
    • Korean Journal of Microbiology
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    • v.55 no.2
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    • pp.87-102
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    • 2019
  • Most chronic wounds persist in the inflammatory phase during wound healing due to the biofilm. Biofilms are resistant to antibiotics, weakening penetration, resistance to biocides and weakening local immune responses. The biofilm is firmly attached to the surrounding tissues and is very difficult to remove. Therefore, strategies to remove hard biofilms without damaging surrounding tissue are very important. One of possible strategies is dispersal. So many studies have been done to develop new strategies using dispersal mechanisms. In this review paper, especially chemotaxis, phage therapy, polysaccharides, various enzymes (glycosidases, proteases, and deoxyribonucleases), surfactants, dispersion signals, autoinducers, inhibitors were introduced. Combination therapies with other therapies such as antibiotic therapy were also introduced. It is expected that the possibility of treatment of chronic wound infection using the knowledge of the biofilm dispersal mechanisms presented in this paper will be higher.

Development of Molecular Marker through Genome Realignment for Specific Detection of Xanthomonas campestris pv. campestris Race 5, a Pathogen of Black Rot Disease

  • Afrin, Khandker Shazia;Rahim, Md Abdur;Jung, Hee-Jeong;Park, Jong-In;Kim, Hoy-Taek;Nou, Ill-Sup
    • Journal of Microbiology and Biotechnology
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    • v.29 no.5
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    • pp.785-793
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    • 2019
  • Black rot caused by Xanthomonas campestris pv. campestris (Xcc) is the most damaging disease in Brassica crops around the world. In this study, we developed a molecular marker specific to Xcc race 5. To do this, the available whole genome sequences of Xcc races/strains and Xc subspecies were aligned and identified a highly variable genomic region (XccR5-89.2). Subsequently, a primer set covering the 'XccR5-89.2' region was designed and tested against the genomic DNA of Xcc races/strains, Xc subspecies and other plant-infecting bacterial strains (Pseudomonas syringae pv. maculicola and Erwinia carotovora subsp. carotovora). The results showed that the 'XccR5-89.2' primer pair amplified a 2,172-bp fragment specific to Xcc race 5. Moreover, they also amplified a 1,515-bp fragment for Xcc race 1 and an over 3,000-bp fragment for Xcc race 3. However, they did not amplify any fragments from the remaining Xcc races/strains, subspecies or other bacterial strains. The 'XccR5-89.2' primer pair was further PCR amplified from race-unknown Xcc strains and ICMP8 was identified as race 5 among nine race-unknown Xcc strains. Further cloning and sequencing of the bands amplified from race 5 and ICMP8 with 'XccR5-89.2' primers revealed both carrying identical sequences. The results showed that the 'XccR5-89.2' marker can effectively and proficiently detect, and identify Xcc race 5 from Xcc races/strains, subspecies and other plant-infecting bacteria. To our knowledge, this is the first report for an Xcc race 5-specific molecular marker.

The Effect of Irradiation on Meat Products

  • Yea-Ji Kim;Ji Yoon Cha;Tae-Kyung Kim;Jae Hoon Lee;Samooel Jung;Yun-Sang Choi
    • Food Science of Animal Resources
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    • v.44 no.4
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    • pp.779-789
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    • 2024
  • The effects of irradiation on meat constituents including water, proteins, and lipids are multifaceted. Irradiation leads to the decomposition of water molecules, resulting in the formation of free radicals that can have both positive and negative effects on meat quality and storage. Although irradiation reduces the number of microorganisms and extends the shelf life of meat by damaging microbial DNA and cell membranes, it can also accelerate the oxidation of lipids and proteins, particularly sulfur-containing amino acids and unsaturated fatty acids. With regard to proteins, irradiation affects both myofibrillar and sarcoplasmic proteins. Myofibrillar proteins, such as actin and myosin, can undergo depolymerization and fragmentation, thereby altering protein solubility and structure. Sarcoplasmic proteins, including myoglobin, undergo structural changes that can alter meat color. Collagen, which is crucial for meat toughness, can undergo an increase in solubility owing to irradiation-induced degradation. The lipid content and composition are also influenced by irradiation, with unsaturated fatty acids being particularly vulnerable to oxidation. This process can lead to changes in the lipid quality and the production of off-odors. However, the effects of irradiation on lipid oxidation may vary depending on factors such as irradiation dose and packaging method. In summary, while irradiation can have beneficial effects, such as microbial reduction and shelf-life extension, it can also lead to changes in meat properties that need to be carefully managed to maintain quality and consumer acceptability.

Gemcitabine-induced Cell Death in Lung Cancer Cells : the Role of p53 (폐암 세포에서 Gemcitabine에 의한 세포 사멸과 p53의 역할)

  • Kim, Doh-Hyung;Bae, Gang-U;Yong, Wha-Shim;Choi, Eun-Kyung;Kim, Youn-Seup;Park, Jae-Seuk;Jee, Young-Koo;Lee, Kye-Young
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.3
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    • pp.275-284
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    • 2002
  • Background : Gemcitabine is a new anti-cancer agent for treating non-small cell lung cancer. Functioning as an antimetabolite, it induces anti-cancer effects by suppressing DNA synthesis after being incorporated into the DNA as a cytosine arabinoside analogue. When Gemcitabine is incorporated into the DNA, the p53 gene may be activated by induction of the DNA defect. However, there are a few studies on the molecular mechanisms of Gemcitabine-induced cell death. This study examined the role of p53 in Gemcitabine-induced cell death. Methods : A549 and NCl-H358 lung cancer cells were used in this study. The cell viability test was done using a MTT assay at Gemcitabine concentrations of 10nM, 100nM, 1uM, 10uM and 100uM. A FACScan analysis with propium iodide staining was used for the cell cycle analysis. Western blot analysis was done to investigate the extent of p53 activation. For the functional knock-out of p53, stable A549-E6 cells and H358-E6 cells were transfected pLXSN-16E6SD which is over expresses the human papilloma virus E6 protein that constantly degrades p53 protein. The functional knock out of p53 was confirmed by Western blot analysis after treatment with a DNA damaging agent, doxorubicine. Results : Gemcitabine exhibited cell toxicity in dose-dependent fashion. The cell cycle analysis resulted in an S phase arrest. Western blot analysis significant p53 activation in time-dependent manner. Gemcitabine-induced cytotoxicity was reduced by 20-30% in the A549-E6 cells and the 30-40% in H358-E6 cells when compared with the A549-neo and H358-neo control cells. Conclusion : Gemcitabine induces an S phase arrest, as expected for the anti-metabolite, and activates the p53 gene, Furthermore, p53 might play an important role in Gemcitabine-induced cell death. Further investigation into the molecular mechanisms on how Gemcitabine activates the p53 gene and its signaling pathway are recommended.

Peripheral Blood Lymphocytes as In Vitro Model to Evaluate Genomic Instability Caused by Low Dose Radiation

  • Tewari, Shikha;Khan, Kainat;Husain, Nuzhat;Rastogi, Madhup;Mishra, Surendra P;Srivastav, Anoop K
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.1773-1777
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    • 2016
  • Diagnostic and therapeutic radiation fields are planned so as to reduce side-effects while maximising the dose to site but effects on healthy tissues are inevitable. Radiation causes strand breaks in DNA of exposed cells which can lead to chromosomal aberrations and cause malfunction and cell death. Several researchers have highlighted the damaging effects of high dose radiation but still there is a lacuna in identifying damage due to low dose radiation used for diagnostic purposes. Blood is an easy resource to study genotoxicity and to estimate the effects of radiation. The micronucleus assay and chromosomal aberration can indicate genetic damage and our present aim was to establish these with lymphocytes in an in vitro model to predict the immediate effects low dose radiation. Blood was collected from healthy individuals and divided into 6 groups with increasing radiation dose i.e., 0Gy, 0.10Gy, 0.25Gy, 0.50Gy, 1Gy and 2Gy. The samples were irradiated in duplicates using a LINAC in the radiation oncology department. Standard protocols were applied for chromosomal aberration and micronucleus assays. Metaphases were stained in Giemsa and 200 were scored per sample for the detection of dicentric or acentric forms. For micronuclei detection, 200 metaphases. Giemsa stained binucleate cells per sample were analysed for any abnormality. The micronuclei (MN) frequency was increased in cells exposed to the entire range of doses (0.1-2Gy) delivered. Controls showed minimal MN formation ($2.0%{\pm}0.05$) with triple MN ($5.6%{\pm}2.0$) frequency at the lowest dose. MN formation increased exponentially with the radiation dose thereafter with a maximum at 2Gy. Significantly elevated numbers of dicentric chromosomes were also observed, even at doses of 0.1-0.5Gy, compared to controls, and acentric chromosomes were apparent at 2Gy. In conclusion we can state that lymphocytes can be effectively used to study direct effect of low dose radiation.

Effect of Cholesterol Supplementation in Freezing Medium on the Survival and Integrity of Human Sperm after Cryopreservation (콜레스테롤이 동결-해동 후 인간정자의 생존과 기능보존에 미치는 영향)

  • Lim, Jung-Jin;Sung, Su-Ye;Kim, Kye-Seong;Song, Seung-Hon;Lee, Woo-Sik;Yoon, Tae-Ki;Lee, Dong-Ryul
    • Clinical and Experimental Reproductive Medicine
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    • v.35 no.3
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    • pp.203-212
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    • 2008
  • Objective: During cryopreservation process, cold shock and cryo-injury affect the fertilizing capacity of the sperm by damaging cell membranes with loss of functional integrity. A longstanding concept for preventing the cryo-damage is to stabilize the plasma membrane by incorporating cholesterol. This study was to determine the effects of cholesterol in freezing media on the motility and functional integrity of human sperm after cryopreservation. Methods: Control group (non-cholesterol treated) and different concentrations of cholesterol-treated sperm (14 healthy males) were frozen and thawed. After freezing and thawing of sperm, the quality of sperm was evaluated by sperm analysis, acrosome reaction test and sperm chromatin structure assay. Results: When human sperm were incubated in sperm freezing medium (SFM) containing $0.5{\mu}g$ cholesterol and then freezing/thawing, the motility of sperm have significantly improved compared to those untreated cholesterol ($33.46{\pm}1.48%$ vs. $30.10{\pm}1.07%$, p<0.05). The rate of calcium ionophore-induced acrosome reactions in post-thawed sperm was significantly higher than that ($53.60{\pm}1.60%$ vs. $47.40{\pm}1.86%$, p<0.05) in SFM containing cholesterol. Sperm chromatin structure assay revealed that DNA damage to the sperm in the cholesterol-treated group was lower than that of non-treated group. Conclusion: These results suggest that increased cholesterol content of sperm plasma membrane by supplementation of cholesterol in SFM improves sperm motility, capacitation status, and DNA integrity. Therefore, addition of cholesterol into SFM could be a useful for protecting human sperm from cold shock and cryo-injury during cryopreservation.

The Stability of p53 in Ras-mediated Senescent Cells in Response to Nucleolar Stress (Ras에 의해 유도된 노화세포에서 핵인 스트레스에 의한 p53 안정화 연구)

  • Sihn, Choong-Ryoul;Park, Gil-Hong;Lee, Kee-Ho;Kim, Sang-Hoon
    • Journal of Life Science
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    • v.19 no.4
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    • pp.436-441
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    • 2009
  • B23/nucleophosmin, a nucleolar protein, translocates into the nucleus from the nucleolus when cells are damaged by extracellular stresses. Recently, it was shown that such translocation of B23/nucleophosmin in normal fibroblasts under stress conditions increases both the stability and activation of the p53 protein by disrupting its interaction with MDM2. Senescent cells have a single large nucleolus and a diminished capacity to induce p53 stability upon exposure to various DNA damaging agents. To investigate the role of B23/nucleophosmin in p53 stability in senescent cells, we established a senescence model system by expressing the ras oncogene in IMR90 cells. The stability of p53 was reduced in these cells in response to nucleolar stress, although the level of B23/nucleophosmin protein was not changed. In addition, p53 did not accumulate in the nucleus and B23/nucleophosmin did not translocate into the nucleoplasm. The binding affinity of B23/nucleophosmin with p53 was reduced in senescent cells, whereas the interaction between MDM2 and p53 was stable. Taken together, the stability of p53 in ras-induced senescent cells may be influenced by the ability of B23/nucleophosmin to interact with p53 in response to nucleolar stress.