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Processing these data using the ΔΔCT method yields the normalized dengue expression ratios presented in Table 2.
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Infected bone marrow cells derived from engrafted NOD-scid IL2rγnull mice were also assessed for dengue antigen expression by immunoperoxidase staining.
During acute Dengue Fever, iNOS expression in monocytes differed among patients probably due to individual variations.
Most attempts to express dengue NS1 in E. coli and yeast expression system have ended up with insoluble protein.
48 hours post-transfection, expression of dengue E protein was monitored by flow cytometry on permeabilized transfected HeLa cells with the 4E11 monoclonal antibody against dengue E protein.
Viral vectors in which this technique has been successfully used to control transgene expression include Dengue virus replicons [12], Alphaviral replicons [13], Lentiviral vectors [14] and Adenoviral vectors [15].
This study, related to gene expression of dengue patients and application of the SVM algorithm to the data is part of a bigger functional immunomics study in our lab and was reviewed and approved by ethics committee of Brazilian Ministry of Health CONEP: 4909; Process n° 25000.119007/2002-03; CEP: 32/09.
Additional clinical information is needed to determine the influence of co-infections on clinical expression of dengue and chikungunya fever.
There are several reports of cloning and expression of dengue NS1 protein for research and diagnostic purpose in a variety of expression systems like bacteria, yeast, and insects.
Functions and molecular mechanisms of MD2-like proteins have not been elucidated fully in mosquitoes and several MD-like proteins have altered gene expression following dengue infection in salivary glands or whole mosquito bodies [ 30, 31].
Our gene expression data from dengue disease patients provides an opportunity to better understand the relevance of the individual genes in the DENV infection pathway involving the innate immune response.
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