Nanji Alcohol Diet Protocol

Both genetic and environmental models of circadian disruption used the Nanji diet alcohol protocol [32] (link), [33] (link), [34] (link), [35] (link), which consisted of a two week introduction and gradual increase in alcohol dose, followed by eight weeks on the full alcohol concentration (29% of total calories, 4.5% v/v; Figure 1). Control mice were fed an isocaloric liquid diet in which the calories from alcohol were replaced with dextrose. The components of the liquid Nanji diet include: mineral mix, vitamin mix, choline bitartrate, d-L-methionine, lactalbumin, xanthan gum, dextrose (all obtained from Dyets, Inc., Bethlehem, PA), fish oil (from menhaden), ethanol (both from Sigma, St. Louis, MO) and Hershey's Lite Chocolate syrup. The caloric composition of the diet was: 36% protein, 29% carbohydrate/alcohol and 35% fat. The liquid diet for each group was prepared fresh daily and provided to mice in individual specialized graduated sipper tubes (Bio-Serv, Frenchtown, NJ) to allow for monitoring of daily food intake.