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5)        The 12 hour overnight fast is very long for a mouse and would not be approved without critique by an IACUC in the US. Did animals receive water access during this time? More information as to why this time point was selected needs to be discussed in the methods. I personally do not believe there is much scientific justification for this lengthy fast as intestinal contents would have been emptied long before 12 hours in a rodent. This will also induce a significant amount of stress in the animal, particularly during the night cycle, which will impact nearly all the assays performed in this study. The authors should review: http://www.norecopa.no/norecopa/vedlegg/food-deprivation.pdf.  While the length of the fast does not render the paper flawed by any means, it just requires additional justification in the methods section.
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