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  • 1
    ISSN: 1432-2277
    Keywords: Key words Reverse transcriptase polymerase chain reaction ; Transplantation ; Heparin ; Ischemia-reperfusion ; Gene expression ; Cytokines
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract The reverse transcriptase polymerase chain reaction (RT-PCR) is a rapid and sensitive method for detecting gene expression. However, when we used this technique to study gene expression of cytokines in ischemic and ex-vivo-reperfused rat lungs as a model for transplantation, significant inhibition of RT-PCR reaction was observed. To optimize RT-PCR conditions, RNA was extracted from rat lungs after flushing, preservation, and reperfusion. RNA was further purified and PCR conditions were modified with various strategies. We found that heparinase I pretreatment completely overcame the inhibitory effects of RT-PCR using RNA extracted from lung tissues after ischemia-reperfusion. With this treatment, a dramatic increase in tumor necrosis factor-α (TNF-α) mRNA was revealed from lung tissues after ischemia-reperfusion. This result suggests that residual heparin in lung tissue interferes with RT-PCR. Because heparinization is routinely used during clinical and experimental organ transplantation, we recommend the treatment of RNA samples with heparinase prior to RT-PCR.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Chichester : Wiley-Blackwell
    International Journal for Numerical Methods in Fluids 17 (1993), S. 221-239 
    ISSN: 0271-2091
    Keywords: Turbulent reacting flows ; Modelling ; Solution sensitivity ; Numerical methods ; Multi-grid method ; Numerical accuracy ; Engineering ; Engineering General
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: An evaluation of some numerical methods for turbulent reacting flows in furnace-like geometries is carried out. The Reynolds averaged Navier-Stokes equations and the two-equation k-∊ model together with either finite-rate or infinite-rate reaction models are solved numerically. Either single- or multiple-step reactions together with the ‘eddy dissipation concept’ (EDC) are used to model reacting flows with finite reaction rates. The numerical scheme is finite difference based, together with a multi-grid method and a local grid refinement technique. These methods have been used to calculate the combustion of propane in a single- and multiple-burner configurations. In the former case, the sensitivity of the solution to variations in some model parameters (determining the reaction rate) and numerical parameters (mesh spacing) has been studied. It is noted that different dependent variables exhibit different levels of sensitivity to the variation in model parameters. Thus, calibration and validation of models for reacting flows require that one compares the most sensitive variables. For engineering purposes, on the other hand, one may calibrate and validate models with respect to the most relevant variables. Our conclusion is that since sensitivity of the temperature distribution is relatively mild, one can still use EDC-like methods in engineering applications where details of the temperature field are of minor importance.
    Additional Material: 13 Ill.
    Type of Medium: Electronic Resource
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