• Title/Summary/Keyword: 독성민감도

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Apoptotic response to various apoptotic inducers on cultured HCE cells (여러 가지 apoptosis 유도 물질의 각막 상피세포에 대한 apoptosis 유도 반응)

  • Kim, Jai-Min;Kim, Soon-Ae;Yoo, Geun-Chang;Seo, Eun-Sun
    • Journal of Korean Ophthalmic Optics Society
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    • v.8 no.1
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    • pp.65-71
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    • 2003
  • The corneal epithelium is constantly being shed. The mechanism of corneal desquamation is not fully understood. Apoptosis, programmed cell death, may play a role. Apoptosis can be induced by a number of factors and different mechanisms. The study was performed to examine the apoptotic index induced in human corneal epithelial cells maintained in tissue culture by various apoptotic inducers. Various inducers, recombinant human cytokines($INF{\gamma}$, $TNF{\alpha}$, FASAb), actinomycin D. camptothecin, cycloheximide, dexamethasone and etoposide, were purchased from commercial suppliers. Inducers at manufacturer-recommended concentration were added to the corneal epithelial cells for 48 hours. Cell viability was measured using MTT assay. The cells were then assessed for the level of apoptosis. Morphologic changes and quantification of apoptotic cells were determined and counted under fluorescence microscope after inducers-treated human corneal epithelial (HCE) cells for 48 hours with Hoechst 33342 staining. Annexin V-FITC/PI staining and DePsipher assay. The expression of Fas protein was studied by immunocytochemistry. All inducers induced apoptosis in HCE cells in a dose dependent manner. Actinomycin D. camptothecin and etoposide induced apoptosis at lower than manufacturer-recommended concentration, while cytokines, cycloheximide and dexamethasone induced apoptosis at higher concentrations at the end of 48 hours. All inducers elicited typical apoptotic morphologic changes (chromatin condensation, nucleus fragmentations non-orange-red colored mitochondria) and expresses Fas protein highly. Apoptotic index of HCE cells by these inducers was different from the other cell lines. RNA synthesis inhibitor and topoisomerase inhibitors induced apoptosis at lower concentration than manufacturer-recommended concentration. Cytokines, cycloheximide and dexamethasone were able to produce apoptosis at 10 times higher concentrations. RNA synthesis inhibitor and topoisomerase inhibitors are more sensitive than intracellular receptor-activators in apoptotic induction of HCE cells.

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Application of a Peptide Nucleic Acid-Based Asymmetric Real-Time PCR Method for Rapid Detection of Vibrio cholerae (비브리오 콜레라 신속 검출을 위한 펩티드 핵산 기반 비대칭 real-time PCR 방법의 적용)

  • Kang, Mingyeong;Lee, Taek-Kyun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.12
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    • pp.117-124
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    • 2019
  • Vibrio cholerae is a very important pathogenic bacterium that has to be monitored in seafood and ships' ballast water. Various methods have been developed to identify this bacterium, yet these methods are time-consuming and have limitations for their sensitivity to detect contamination. The purpose of the present study was to develop a robust and reliable method for identifying V. cholerae. Peptide nucleic acid (PNA) probes were developed to use for PNA-based asymmetrical real-time PCR techniques. The toxigenic Cholera enterotoxin subunit B (ctxB) gene was selected as a target for detecting V. cholerae and the gene was synthesized as a positive template for conventional and real-time PCR. Real-time PCR primers and PNA probes were designed and standard curves were produced for the quantitative analysis. The selected PNA probes reacted specifically to V. cholerae without any ambiguity, even among closely related species, and the detection limit was 0.1 cfu/100 mL. Taken together, the PNA probes and asymmetrical qPCR methods developed in this present study could contribute to the rapid, accurate monitoring of V. cholerae in marine environments, and as well as in seafood and ships' ballast waters.

Removal of Aqueous Cyanide through the Utilization of Industrial By-products (산업부산물(제강 Sludge, 제강 Slag)을 이용한 시안 흡착 제거)

  • 이정원;현재혁;조재범
    • Journal of Korea Soil Environment Society
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    • v.3 no.2
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    • pp.89-99
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    • 1998
  • Sorption of the aqueous cyanide onto steel mill sludge and steel mill slag, both of which are the by-products from the converter furnace, was studied. In the study, the influence of temperature, activation energy, concentration and pH on sorption of cyanide was investigated. Three different temperature($25^{\circ}C$ > $37^{\circ}C$> $50^{\circ}C$) was chosen to represent that of landfill leachate. Initial concentration was 1 mg/$\ell$ 5 mg/$\ell$, 10 mg/$\ell$, and 20 mg/$\ell$. In addition, pH was set to three different level, that is, 3, 7, and 11 respectively. As the result of batch mode experiment for cyanide adsorption, the removal rate was found to be proportional to the initial concentration of cyanide. The order of removal rate was 20 mg/$\ell$> 10 mg/$\ell$> 5 mg/$\ell$> 1 mg/$\ell$. Similarly the influence of pH was proportional because of the change in solubility of cyanide. The order of removal rate was pH 11 > pH 7 > pH 3. As the temperature increased, so did the removal rate. The reaction was endothermic and the value of activation energy(Ea) was 127.93 J/mole and 59.44 J/mole respectively at 1 mg/ιand 20 mg/ιof initial concentration. From the experiment, it can be postulated that the capability of steel mill by-products to attenuate aqueous cyanide is enough to be used as substitute for clay liner of landfill site in the aspect of pollutant removal.

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Distribution of Cyanotoxin Microcystin-LR in Han River System and Ecological Park in Seoul and Kyunggi Districts (서울 경기지역의 공원 연못 및 한강 수계내 조류독소 Microcystin-LR의 분포)

  • Suh, Mi-Yeon;Kim, Baik-Ho;Han, Myung-Soo
    • Korean Journal of Ecology and Environment
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    • v.38 no.2 s.112
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    • pp.237-248
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    • 2005
  • To determine the content of hazard microcystin (MC) in Han River system and Ecological Ponds in Seoul City and Kyunggi district, a most toxic derivative, microcystin-LR (MCLR) of 15 samples of 7 ponds, 4 rivers and 4 reservoirs during low precipitation and cold season in 2003 were analyzed by ELISA method. With the change of water temperature ($0.4\;{\sim}\;21.9^{\circ}C$), cyanobacteria including Microcystis aeruginosa dominated the cold phytoplankton community in small ecological ponds such as Kyungbokgung Kyunghyaeru (KBP), Seokchon reservoir (SCR), Yangsoori Ryukgakji (YSS), having the long residence time. Contents of MCLR (the detection limit; $0.05\;{\mu}g\;L^{-1}$) were high in cyanobacteria-rich sites, especially, Microcystis aeruginosa. Total MCLR, cell extracted type plus dissolved type, were $1.39\;{\mu}g\;L^{-1}$ in KBP, $0.55\;{\mu}g\;L^{-1}$ in SCR and $0.59\;{\mu}g\;L^{-1}$ in YSS, in the first sampling having a high temperature (>$20^{\circ}C$), while some detected only in YSS during the cold season. As expected, the MCLR content was correlated with Microcystis aeruginosa (r = 0.526 for cell extracted type, r = 0.433 for dissolved type). Therefore, low concentration of MCLR detected in small ponds and Han river system in Seoul metropolitan city and Kyunggi district, maybe hardly affect human recreation activity, especially the drinking water supply.

Biochemical Aspect of Superoxide Toxicity to Plant Mitochondria (식물 미토콘드리아에 대한 Superoxide독성의 생화학적 측면)

  • Jung, Jin;In, Man-Jin
    • Applied Biological Chemistry
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    • v.32 no.1
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    • pp.23-29
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    • 1989
  • Biochemical consequence of the accumulation in cells of superoxide $(O^{-}_{2})$ which was proposed to be probably a common chemical factor in the secondary process of the mechanism of chilling injury as well as in the visible light photodamage in cells of higher plants, has been investigated in the present work. Especially focused was the destructive effect of $O^{-}_{2}$ on the biochemical activity of mitochondria, as informations which support the suggestion that mitochondrial inner membrane is the major site of $O^{-}_{2}$ production have been collected. Mitochondria and submitochondrial particles (SMP) were prepared from soybean hypocotyls for this case study. When SMP were treated with the electrolytically produced $O^{-}_{2}$ they suffered not only inhibition of the membrane-bound enzymes as demonstrated by cytochrome c oxidase, but also lipid peroxidation of membrane as proved by malondialdehyde production. Malate dehydrogenase present in the protein extract from mitochondrial matrix was also inhibited by the $O^{-}_{2}$ treatment. These results exhibited the chaotic effect of the overproduction and accumulation of $O^{-}_{2}$ in cells under a certain abnormal circumstance such as environmental stress on the physiological function of mitochondrial; disruption of the cellular metabolic pathways and the structural integrity of membrane.

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Evaluation of DNA Damage and Repair Kinetics in the Earthworm (Eisenia fetida) Exposed to Radiation and Mercury (방사선과 수은에 의해 유도된 Eisenia fetida 체강세포의 DNA 손상 및 수복 평가)

  • Ryu, Tae-Ho;Nili, Mohammad;An, Kwang-Guk;Kim, Jin-Kyu
    • Korean Journal of Environmental Biology
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    • v.29 no.1
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    • pp.68-73
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    • 2011
  • The single cell gel electrophoresis (SCGE) assay is a microelectrophoretic technique for assessments of DNA damage at the level of the individual eukaryotic cell. The SCGE assay, due to its simplicity, sensitivity and need of a few cells, has advantages compared to other genomic damage assays such as sister chromatid exchange, chromosomal aberration and micronucleus test. In this study, investigated were the levels of DNA damage and the repair kinetics in the coelomocytes of Eisenia fetida treated with HgCl2 and ionizing radiation by means of the SCGE assay. For detecting DNA damage and repair in coelomocytes, earthworms (E. fetida) were irradiated with six doses of ${\gamma}$-rays (0, 2.5, 5, 10, 20 and 50 Gy) and in vivo exposed to mercuric chloride at 0, 80 and 160 mg $kg^{-1}$ for 48 hours. Then the Olive tail moments were measured during 0~12 hours after irradiation and 0~72 hours after Hg treatment. The results showed that the more the oxidative stress was induced by mercury and radiation, the longer the repair time was required. Also, the results suggest that the SCGE assay may be used as an important tool for comparison of the sensitivity of different species to oxidative stresses.

Apoptotic Response of Human Oral Squamous Carcinoma Cells to Etoposide (Etoposide에 대한 사람구강편평상피암종세포의 세포자멸사 반응)

  • Kim, Gyoo-Cheon;Lee, Kyoung-Duk;Park, Jae-Hyun;Kim, Duk-Han;Park, Jeong-Kil;Park, June-Sang;Park, Bong-Soo
    • Journal of Oral Medicine and Pain
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    • v.30 no.2
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    • pp.231-238
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    • 2005
  • Anti-cancer drugs have been shown to target diverse cellular functions in mediation cell death in chemosensitive tumors. Most antineoplastic drugs used in chemotherapy of leukemias and solid tumors induce apoptosis in drug-sensitive target cells. However, the precise molecular requirements that are central for drug-induced cell death are largely unknown. Etoposide is used for the treatment of lung and testicular cancer. This study was performed to examine whether etoposide promote apoptosis in human oral squamous carcinoma cells (OSC9) as well as in lung and testicular cancer. Etoposide had a significant dose- and time-dependent inhibitory effect on the viability of OSC9 cells. TUNEL assay showed the positive reaction on condensed nuclei. Hoechst stain demonstrated that etoposide induced a change in nuclear morphology. The expression of p53 was increased at 48 hour, suggesting that the nuclear of OSC9 cell was damaged, thereby inducing apoptosis. Etoposide treatment induced caspase-3 cleavage and activation. Intact PARP protein 116-kDa and 85-kDa cleaved product were observed. The activated caspase-3 led cleavage of the PARP. These results demonstrate that etoposide-induced apoptosis in OSC9 cells is associated with caspase-3 activation.

Effects of Cadmium Treatment on Carbohydrate Allocation and Partitioning of Five Half-sib Families of Betula schmidtii (박달나무 반형매 5가계의 탄수화물 배분 및 분할에 대한 Cd 처리 효과)

  • Han, Sim-Hee;Oh, Chang-Young;Lee, Jae-Cheon;Kim, Pan-Gi
    • Korean Journal of Agricultural and Forest Meteorology
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    • v.8 no.1
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    • pp.15-21
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    • 2006
  • This study was conducted to understand the regulation mechanism of carbohydrate allocation and partitioning of one-year-old seedlings of Betula schmidtii with Cd treatment, and to assess tolerance against Cd toxicity among five half-sib families on the basis of carbon allocation and partitioning. Seedlings were treated with $CdSO_4$ solution of 0, 0.2, 0.4, and 0.6 mM for two months. After harvesting, carbohydrate concentrations were analyzed for leaves, stems and roots of seedlings. Carbohydrate concentration for Cd-treated seedlings decreased in comparison with control plants, even though Cd effects were significantly different among five families. Meanwhile, Cd treatment decreased carbohydrate allocation in leaves and increased allocation in roots. In addition, partitioning of glucose in leaves was increased by Cd treatment, but partitioning of sucrose and starch in leaves decreased. In Cd-treated roots, partitioning of glucose, sucrose and starch increased. On the basis of carbohydrate allocation patterns, 'Family No. 7' (of the five families studied) was considered as the most sensitive family to Cd toxicity because the decrease of carbohydrate concentration and the change of carbohydrate allocation pattern after Cd treatment were relatively greater.

Glyphosate Toxicity: II. EPSP-synthase Activity in Cell Suspension Culture of Corydalis Sempervirens and Lycopersicon Esculentum (Glyphosate 독성(毒性): II. corydalis Sempervirens와 토마토의 세포배양체(細胞培養體)에서 EPSP-synthase의 활성(活性)에 미치는 영향(影響))

  • Kim, Tae-Wan;Amrhein, Nikolaus
    • Korean Journal of Weed Science
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    • v.15 no.2
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    • pp.148-153
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    • 1995
  • Glyphosate (N-[phosphonomethyl]glycine) applied to the assimilate-exporting leaves or sprayed to the whole plants of tomato(Lycopersicon esculentum Mil var. Moneymaker) induced the rapid inhibition of 5-enolpyruvyl skimic acid 3-phosphate synthase(EPSP-synthase). It shows that EPSP-synthase activity precedes chlorophyll loss. There is no difference in EPSP-synthase activity between in vivo tomato meristem and cell suspension culture if glyphosate is not applied. The EPSP-synthase activity is in a range of 4 to 6 nkat per mg protein. The inhibition of EPSP-synthase action is induced within 36 h after glyphosate application while the Chl contents were reduced 48 h after the application. In cell suspension culture of tomato and Corydalis (Corydalis sempervirens), a sublethal concentration of glyphosate retards the fresh weight increase and prolonged lag phase. The fresh weight is reached maximal about 14 days after the subculture in the presence of glyphosate. The inhibitory effect of glyphosate on EPSP-synthase is remarkably induced in lag phase.

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Toxins and Antibiotic Resistance of Methicillin-Resistant Staphylococcus aureus Isolated from Clinical Specimens (임상검체로부터 분리된 methicillin 내성 Staphylococcus aureus의 독소 및 항생제 내성)

  • Baik, Keun-Sik;Ki, Gwang-Seo;Choe, Han-Na;Park, Seong-Chan;Koh, Eun-Cho;Kim, Hyung-Rak;Seong, Chi-Nam
    • Journal of Life Science
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    • v.21 no.2
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    • pp.257-264
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    • 2011
  • Seventy five methicillin- resistant Staphylococcus aureus (MRSA) strains and 24 methicillin- susceptible S. aureus (MSSA) were isolated from clinical specimens obtained from a hospital in Suncheon, Jeonnam province, Korea, from July to December, 2009. Antibiotic resistance was determined using the disc diffusion method. Genes encoding enterotoxin (SE), toxic shock syndrome toxin-1 (TSST-1), exfoliative toxin (ET) and Panton-Valentine leukocidin (PVL) were detected by multiplex PCR-mediated amplification using specific primers. Sixty (80%) MRSA isolates possessed either one or more toxin genes and the most common pattern that coexisted in MRSA was seb, sec, seg, sei and tst (22.7%) followed by coexistence of sec, seg, sei and tst genes (18.7%). Gene pvl encoding leukocidin was not found. Significant correlation between the production of sec, seg, sei and tst genes was found. MRSAs were resistant to erythromycin (89% of the isolates), gentamicin (70.7%), ciprofloxacin (69.3%), clindamycin (61.3%) and tetracycline (58.7%), while MSSAs were susceptible to the antibiotics with the exception of erythromycin. Toxin genes seb, sec and tst were related to the tetracycline resistance of MRSA.