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Food intake assay was modified from [53].
Moreover, by using a food intake assay, such as feeding with colored yeast (data not shown), analyzing metabolic markers (supplementary material Fig. S3A) or quantifying mouth hook contractions (supplementary material Fig. S3B), we could exclude the hypothesis that the smaller sizes of dGPHR larvae reflect the inability of dGPHR larvae to feed properly.
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Moreover, sick animals eat and drink less, which can bias measures of sweetened fluid intake in assays designed to evaluate anhedonia (the inability to experience pleasure from naturally rewarding activities).
Then, we performed Western blot, I intake and efflux assays, and clonogenic assay with cancer cells.
However, Ddc-GAL4 and control flies had similar consumption, measured with both a colorimetric method and a dynamic measurement of food intake, the CAFE assay (Ja et al. 2007).
There are many potential errors in biomarker analysis related to sampling, biological variations (eg, seasonal, diurnal and food intake), analyte features, assay format and parameters, 19 which can confound results.
To test this hypothesis, we measured total dietary intake employing the CAFE assay [14].
The observed increase in quinine intake in the CAFE assay was paralleled by a substantial increase in the proboscis extension response.
Although this is a less direct measure of food intake than the CAFE assay and requires two additional controls (Table S1), it can be performed in dietary conditions more comparable to those in which the flies were bred.
To assess food intake, we employed the CAFE assay [14], which quantifies dietary intake using a volumetric capillary feeder.
It was highlighted that validation of such a biomarker requires demonstration of (a) assay robustness, (b) intake versus biomarker level, and (c) stability of stored samples.
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