The changes of DHN1 expression and subcellular distribution in A. delicisoa cells under osmotic stress were studied by using GFP as a reporter molecule. Through creating the Xba I and BamH I restriction sites at the e...The changes of DHN1 expression and subcellular distribution in A. delicisoa cells under osmotic stress were studied by using GFP as a reporter molecule. Through creating the Xba I and BamH I restriction sites at the ends of dhn1 by PCR, the expression vector for the fusion protein DHN1-mGFP4 was constructed by cloning dhn1 into plasmid pBIN-35SmGFP4. Then the DHN1-mGFP4 expression vector was transformed into A. delicisoa suspension cells by microprojectile bombardment method. Bright green fluorescence of GFP which shows the high-level expression of DHN1-mGFP4 was visualized after culture for 10 h. However, the green fluorescence was only located within the nucleus. By increasing the culture medium osmotic potential, the green fluorescence was visualized in the cytoplasm (mainly around the plasma membranes). The generation of GFP fluorescence in the cytoplasm was also promoted by increasing the medium osmotic potential. Moreover, GFP green fluorescence was abolished by protein synthesis inhibitor dicyclohexylcarbodiimid, indicating that the cytoplasmic DHN1 was newly synthesized under osmotic stress. Furthermore, ABA promoted the presence of green fluorescence in the cytoplasm, and the GFP fluorescence was visualized within a shorter time under a higher osmotic potential.展开更多
The aim of this paper is to present works performed in HTC （Heat-Tech Center）, Research ＆ Development Centre of Veolia Group located in Warsaw regarding assessment of probability of failure in DHN （district heatin...The aim of this paper is to present works performed in HTC （Heat-Tech Center）, Research ＆ Development Centre of Veolia Group located in Warsaw regarding assessment of probability of failure in DHN （district heating network）. This work is a part of a project dedicated to develop a software which objective is to increase reliability of DHN. The research methods consisted of three approaches. First, using database of failures which happened in Warsaw DHN and repairing protocols from past 10 years, a statistics approach was applied to perform first analysis. The result was that pipelines with nominal diameter DN （nominal diameter） ≤ 150 had higher failure rate per km, than pipelines with DN 〉 150. The next step of research was to study influence of internal （corrosion caused by heat carrier, quality of materials） and external （stray currents） factor in order to assess its individual influence on failure rate of pipe and explain reasons of differences in failure rate. To end a FMEA （failure mode and＇effects analysis） will aim to identify the main failures modes appearing on DHN, to estimate the main causes of these failures and to propose the best solutions regarding the causes, the costs and the means available.展开更多
基金This work was supported by the Major State Basic Research Development Program of the People's Republic of China, the National Natural Science Foundation of China (Grant No. 39770077) the Science and Technology Program of the Ministry of Education of
文摘The changes of DHN1 expression and subcellular distribution in A. delicisoa cells under osmotic stress were studied by using GFP as a reporter molecule. Through creating the Xba I and BamH I restriction sites at the ends of dhn1 by PCR, the expression vector for the fusion protein DHN1-mGFP4 was constructed by cloning dhn1 into plasmid pBIN-35SmGFP4. Then the DHN1-mGFP4 expression vector was transformed into A. delicisoa suspension cells by microprojectile bombardment method. Bright green fluorescence of GFP which shows the high-level expression of DHN1-mGFP4 was visualized after culture for 10 h. However, the green fluorescence was only located within the nucleus. By increasing the culture medium osmotic potential, the green fluorescence was visualized in the cytoplasm (mainly around the plasma membranes). The generation of GFP fluorescence in the cytoplasm was also promoted by increasing the medium osmotic potential. Moreover, GFP green fluorescence was abolished by protein synthesis inhibitor dicyclohexylcarbodiimid, indicating that the cytoplasmic DHN1 was newly synthesized under osmotic stress. Furthermore, ABA promoted the presence of green fluorescence in the cytoplasm, and the GFP fluorescence was visualized within a shorter time under a higher osmotic potential.
文摘The aim of this paper is to present works performed in HTC （Heat-Tech Center）, Research ＆ Development Centre of Veolia Group located in Warsaw regarding assessment of probability of failure in DHN （district heating network）. This work is a part of a project dedicated to develop a software which objective is to increase reliability of DHN. The research methods consisted of three approaches. First, using database of failures which happened in Warsaw DHN and repairing protocols from past 10 years, a statistics approach was applied to perform first analysis. The result was that pipelines with nominal diameter DN （nominal diameter） ≤ 150 had higher failure rate per km, than pipelines with DN 〉 150. The next step of research was to study influence of internal （corrosion caused by heat carrier, quality of materials） and external （stray currents） factor in order to assess its individual influence on failure rate of pipe and explain reasons of differences in failure rate. To end a FMEA （failure mode and＇effects analysis） will aim to identify the main failures modes appearing on DHN, to estimate the main causes of these failures and to propose the best solutions regarding the causes, the costs and the means available.
