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RAW264.7 세포주를 이용한 수은화합물의 세포독성기전에 관한 고찰
김공호,고대하,소병율 大韓産業醫學會 1996 대한직업환경의학회지 Vol.8 No.3
Balb/c 마우스의 복강내에 Abelson leukemia virus(A-MuLV)를 주입하여 발생시킨 종양의 복수에서 기원한 RAW264.7 cell line을 배양하는 조건에 여러농도의 수은을 첨가하여 nitrite와 ATP 생성의 변화를 관찰한 결과는 다음과 같다. 수은화합물을 첨가한 배양조건에서 RAW264.7세포의 생존률은 mercury chloride의 경우는 0.8μM이하, methyl mercury chloride의 경우 0.4μM이하의 농도에서는 95% 이상으로 유지되었으며, 그 이상의 농도에서는 생존률이 현저히 감소되었다. 이때 ATP 및 nitrite의 생성능은 수은화합물의 농도증가에 따라 용량의존적으로 감소되었다. 배양액(DMEM)에 4.5g/ℓ의 glucose를 1배에서 5배를 추가하여 농도를 강화시킨 조건에서 RAW264.7 세포를 48시간동안 배양하면 ATP 및 nitrite 생성량은 전반적으로 1배 추가시 가장 많은 양이 생성되었고 배양액의 pH는 1배 추가시 6.7-6.8의 범위로 감소하였다가 glucose 농도 증가에 따라 대조군과 비슷한 7.2까지 증가하였다. 이때 세포의 생존률은 모든 군에서 95%이상으로 유지되었다. 그리고 배양액의 glucose 농도를 2배로 강화시킨 실험조건(배양액내 glucose 기준농도의 1배를 더 첨가한 경우)에서는 mercury chloride 및 methylmercury chloride의 농도를 증가시켰음에도 불구하고 ATP 및 nitrite 생성량은 감소되지 않고 대조군과 비슷한 값으로 유지되었다. 이상의 결과는 수은이 면역세포의 nitric oxide 생성을 저해하는 것은 세포내 ATP생성과 관련한 대사과정, 특히 구연산회로(citric acid cycle)의 억제를 통해 발휘된다는 것을 시사하고 있다. The effects of glucose on the productions of ATP and nitrite which are inhibited by mercury compounds, were examined in a cell culture system of RAW 264.7 cells. The cells were cultured in Dulbecco's Modified Eagle's medium(DMEM) with cytokines, IL-1 and TNF for 24 hours. The viability of RAW 264.7 cells at the end of culture was significantly decreased by mercury chloride or methylmercury chloride added into the media in dose-dependent manner, however the viability of RAW 264.7 cells were influenced in the concentrations less than 0.8μM of mercury chloride or 0.4μM of methylmercury chloride. The addition of 4.5g/l glucose to normal DMEM lowered the pH of media to the range of 6.7-6.8 after 48 hours of culture, but not for the cell survivals. This supplement of glucose to the media also prevented the inhibitions of ATP and nitrite syntheses which were caused by mercurial compounds. These results suggest that the disorder of cell mediated immunity by mercurials could be related to the inhibition of nitric oxide synthesis which seems to be caused by the inhibition of ATP synthesis, especially related to the citric acid cycle.
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수은화합물 및 셀레늄이 EMT-6 세포에 미치는 면역독성학적 영향
소병율,고대하,염정호 大韓産業醫學會 1997 대한직업환경의학회지 Vol.9 No.3
The effects of selenium on the productions of ATP and nitrite which are inhibited by mercurial compounds, were examined in a cell culture system of EMT-6 cells. The cells were cultured in DMEM with cytokines, IL-1 and IFN-γ for 24 hours. The NO₂production of EMT-6 cells at the end of culture was significantly decreased by HgCl₂or CH₃HgCl added into the media in dose-dependent manner, and the viability of EMT-6 cells were decreased lower than 90% in the concentrations≥2.0μM HgCl₂or ≥1.0 μM CH₃HgCl. Simultaneous addition of the equimolar dose of selenium significantly increased synthesis of NO₂and ATP which were inhibited by mercurial compounds. But the single addition of selenium influenced neither the viability of cells nor the productions of NO₂and ATP. These results suggest that the disorder of cell mediated immunity by mercurials could be related to the inhibition of nitric oxide synthesis which seems to be caused by the inhibition of ATP synthesis. Administration of selenium with either HgCl₂or CH₃HgCl protected the cells against the cytotoxicity of mercurials. The protective effect of selenium on cytotoxicity of mercury is due to the formation of inactive complex of Se-Hg.
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Phase determination of a homogentisate dioxygenase from Comamonas sp. strain P19
박석열,소병율,채종찬,김정선 한국구조생물학회 2017 Biodesign Vol.5 No.3
In Comamonas sp. strain P19, homogentisate 1,2-dioxygenase (HGO) catalyzes the conversion of homogentisate to4-maleylacetoacetate by aromatic ring scission, in the breakdown of tyrosine and phenylalanine. To determine themolecular background of the enzymatic mechanism of HGO in this zinc-resistant organism, hmgA encoding HGO ofComamonas sp. strain P19 was cloned, and the expressed protein was purified. The protein was crystallized in solutions I[25% (w/v) polyethylene glycol 3350 and 0.1 M trisodium citrate at pH 5.6] and II [1.4 M ammonium tartrate and 0.1 M bis-Tris at pH 5.5]. X-ray diffraction data were collected to 1.8 Å resolution using synchrotron radiation. The crystal belongs tothe orthorhombic space group P212121, with unit cell dimensions of a = 73.2 Å, b = 100.0 Å, and c = 134.9 Å. A traceableelectron density map was calculated using anomalous diffraction data obtained from a crystal soaked in zinc ions.
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Bacillus subtilis S54 균주를 이용한 고추 역병과 탄저병의 생물학적 방제
이건웅,김명준,박준식,채종찬,이귀재,소병율 한국식물병리학회 2011 식물병연구 Vol.17 No.1
Phytophthora blight and anthracnose disease caused by Phytophthora capsici and Collectotrichum gloeosporioides are the most important devastating diseases of red pepper plants, worldwide. Five different bacterial isolates were isolated from the red pepper rhizosphere and non-rhizosphere soil and subsequently tested for antagonistic activity against P. capsisi and C. gloeosporioides. The area of the inhibition zone was taken as a measure for antagonistic activity. Among the 5 isolates tested, S54 exhibited a maximum antagonistic activity under in vitro and in vivo conditions. In greenhouse studies the isolate has successfully reduced the disease symptom. Protect value was 80.8% (Phytophthora blight) and 81.9% (Anthrancnose disease), whereas the infection rate of control plants was 21.3% and 23.2%. Based on the 16S rDNA sequence and API 50CHB Kit analysis the most effective isolate was identified as Bacillus subtilis. The results of the study indicate that the stratin S54 could be used as an potential biological control of Phytophthora blight and anthracnose disease of red pepper. 초록고추 역병과 탄저병은 Phytophthora capsici와 Colletotrichum gloeosporioides에 의해 발병되는 병으로 세계적으로 많은 피해를 입히는 중요한 병원균이다. 이러한 병원균의 방제를 위하여 근권과 비근권에서 각각의 병원균 생장을 억제하는 능력이 뛰어난 5균주를 선발하였다. 선발된 분리주 중 S54균주는 in vitro와 in vivo 조건 모두에서 가장뛰어나게 병원균을 억제하였다. 하우스 실험에서도 역병과 탄저병 모두 80% 이상의 방제력을 보여주었다. 또한 S54균주의 동정을 위하여 16S rDNA 부분의 염기서열의 분석과 API 50CHB Kit를 실시하였으며, 그 결과 Bacillus subtilis로 동정하였다. 이러한 결과를 바탕으로 S54는 고추 역병과 탄저병을 방제할 수 있는 생물학적 방제제로의 가능성을 보여주었다고 할 수 있다.
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