• Title/Summary/Keyword: In situ RNA hybridization

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Seasonal Variations of Nitrifying Bacteria in Agricultural Reservoir (농업용 저수지에서의 질화세균의 계절적인 변화)

  • Lee, Hee-Soon;Lee, Young-Ok
    • Korean Journal of Ecology and Environment
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    • v.35 no.3 s.99
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    • pp.152-159
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    • 2002
  • The seasonal variations of nitrifying bacterial population sampled from 3 sites in Moon-Chon reservoir were analyzed by in situ hybridization with fluorescently labeled rRNA-targeted oligonucleotide probes from August 2000 until July 2001. In addition, physico-chemical parameters such as temperature, pH, chi-a and DOC were measured to determine correlations between those factors and the size of nitrifying bacterial populations. Total bacterial numbers varied in the range of $0.8{\sim}1.5{\times}10^6\;cells/ml$ independent of sites and had the maximal values in March at all 3 stations. The ratio of eubacteria to total bacteria ranged from 44.9% to 79.5%, and the ratio of each nitrifying bacteria to eubacterial numbers reached only $1.0{\sim}7.4%$. The variations of ammonia-oxidizing bacteria ranged from $1.1{\times}10^4$ to $3.0{\times}10^4\;cells/ml$ without noticeable peak values whereas those of nitrite-oxidizing bacteria varied in $1.3{\sim}5.7{\times}10^4\;cells/ml$ with the increasing tendency in winter regardless of the sites. Moreover it was observed that the numbers of nitrite-oxidizing bacteria were higher than those of ammonia-oxidizing bacteria. Total bacterial numbers correlated with water temperature (r = 0.355, p<0.05) and DOC (r = 0.58G, p<0.01) positively whereas nitrite-oxidizing bacteria correlated with temperature (r = -0.416, p<0.05) and pH (r = -0.568, p = 0.001) negatively. In addition, DOC represented good correlations with eubacterial numbers (r = 0.448, p<0.01). These results indicate that temperature, DOC and pH might be one of the main factors affecting variations of bacterial populations in the aquatic ecosystem. It was also suggested that FISH method is a useful tool for detection of slow growing nitrifying bacteria.

Role of Growth Factors and Cytokines on Bleomycin Induced Pulmonary Fibrosis (Bleomycin 유도 폐 섬유화에 있어서 성장인자 및 Cytokine의 역할)

  • Lee, Yong-Hee;Jung, Soon-Hee;Ahn, Chul-Min;Kim, Sung-Kyu;Cho, Sang-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.44 no.4
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    • pp.871-888
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    • 1997
  • Background : It is now thought that the earliest manifestation of idiopathic pulmonary fibrosis is alveolitis, that is, an accumulation of inflammatory and immune effector cells within alveolar walls and spaces. Inflammatory cells including alveolar macrophages and resident normal pulmonary tissue cells participate through the release of many variable mediators such as inflammatory growth factors and cytokines, which contribute to tissue damage and finally cause chronic pulmonary inflammation and fibrosis. This study was performed to investigate the source and distribution pattern of transforming growth factor-${\beta}_1$(TGF-${\beta}_1$), platelet derived growth factor(PDGF), basic fibroblast growth factor(bFGF), interleukin 1(IL-1), interleukin 6(IL-6), tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and the role of these mediators on bleomycin(BLM)-induced pulmonary injury and fibrosis in rats. Method : Wistar rats were divided into three groups(control group, BLM treated group, BLM and vitamine E treated group). Animals were sacrificed periodically at 1, 2, 3, 4, 5, 7, 14, 21, 28 days after saline or BLM administration. The effects were compared to the results of bronchoalveolar lavage fluid analysis, light microscopic findings, immunohistochemical stains for six different mediators(TGF-${\beta}_1$, PDGF, bFGF, IL-1, IL-6 and TNF-$\alpha$) and mRNA in situ hybridization for TGF-${\beta}_1$. Results : IL-1 and IL-6 are maximally expressed at postbleomycin 1~7th day which are mainly produced by neutrophils and bronchiolar epithelium. It is thought that they induce recruitment of inflammatory cells at the injury site. The expression of IL-1 and IL-6 at the bronchiolar epithelium within 7th day is an indirect evidence of contribution of bronchiolar epithelial cells to promote and maintain the inflammatory and immune responses adjacent to the airways. TNF-$\alpha$ is mainly produced by neutrophils and bronchiolar epithelial cells during 1~5th day, alveolar macrophages during 7~28th day. At the earlier period, TNF-$\alpha$ causes recruitment of inflammatory cells at the injury site and later stimulates pulmonary fibrosis. The main secreting cells of TGF-${\beta}_1$ are alveolar macrophages and bronchiolar epithelium and the target is pulmonary fibroblasts and extracellular matrix. TGF-${\beta}_1$ and PDGF stimulate proliferation of pulmonary fibroblasts and TGF-${\beta}_1$ and bFGF incite the fibroblasts to produce extracellular matrix. The vitamine E and BLM treated group shows few positive cells(p<0.05). Conclusion : After endothelial and epithelial injury, the neutrophils and bronchiolar epithelium secrete IL-1, IL-6, TNF-$\alpha$ which induce infiltration of many neutrophils. It is thought that variable enzymes and $O_2$ radicals released by these neutrophils cause destruction of normal lung architecture and progression of pulmonary fibrosis. At the 7~28th day, TGF-${\beta}_1$, PDGF, bFGF, TNF-$\alpha$ secreted by alveolar macrophages sting pulmonary fibroblasts into proliferating with increased production of extracellular matrix and finally, they make progression of pulmonary fibrosis. TNF-$\alpha$ compares quite important with TGF-${\beta}_1$ to cause pulmonary fibrosis. Vitamine E seems to decrease the extent of BLM induced pulmonary fibrosis.

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