Vanilla ensure

Beginning after day 30 post-surgery, ad libitum-fed mice in caspase-induced genetic ablation experiments were habituated to test cages with fluid spouts equipped with lickometers. After 3 daily 1-h habituation sessions to a spout filled with water, a 1-h test session was conducted in which a caloric liquid (vanilla Ensure; Abbott Laboratories, IL, USA) was freely available and the total number of licks was recorded.

Animals were fasted the night prior to surgery. SG and sham surgeries were performed as previously described including pre-operative antibiotic prophylaxis with enrofloxacin (10 mg/kg subcutaneously), pre-operative pain control with subcutaneous Buprenorphine SR (0.5 mg/kg), and subcutaneous saline (20 cc/kg) on post-operative day 0 and 1 [8 (link)]. In brief, SG was performed by dividing and removing all the non-glandular stomach and glandular lateral fundus using an endoscopic stapling device (Endopath ETS-Flex, 45 mm white load; Ethicon, Cincinnati, OH). For sham animals, the same procedure was followed except the stomach was not divided. Rats received Vanilla Ensure for 48 hours after surgery (Abbott, Lake Forest, IL) before resuming their post-operative diet.

Solid food was replaced with Vanilla Ensure (Abbott, Lake Forest, IL) for post-operative recovery starting 24 hours prior to surgery and for 72 hours after surgery. All animals were fasted overnight but had ad libitum access to water. SG and pair-fed sham, or ad-lib sham surgeries were performed as previously described [5 (link),10 ]. All animals were started on Purina diet 5008 72 hours after surgery. SG and ad-lib sham rats have unlimited access to food. Pair-fed sham rats’ food intake was restricted to match the daily food intake of SG rats. Pair-fed sham rats were given the same amount of daily food consumed on average by the SG rats for the same postoperative day throughout the study period. For example, if the average food intake of obese SG rats was 15g on postoperative day #15, all pair-fed sham rats would receive only 15 g of food for their respective postoperative day 15. This creates a pair-fed group that is weight-matched and food-intake matched to the SG group.

Ethanol and control diets were composed of 30% v/v Vanilla Ensure® (Abbott Nutrition, Columbus, Ohio) and an equal quantity of double-distilled water. The ethanol-containing diet was composed of 40% v/v 200 proof ethanol (Sigma-Aldrich, San Diego, CA). Control diets were isocalorically matched to ethanol-containing diets via substitution of maltose (Sigma-Aldrich) dissolved in double-distilled water yielding equivalent caloric intake. In order to determine how many withdrawals were required to produce reliable behavioral manifestations of withdrawal, animals received ethanol (4 g/kg/i.g.) or an isocaloric-containing control diet twice daily at 0800 and 1600 for 5 consecutive days followed by a 2-day period of withdrawal. This regimen was repeated either one, two, or three times (i.e., 1, 2 CIE, or 3 CIE). This study aimed to achieve peak blood ethanol levels (BELs) of approximately 200 mg/dl to reflect patterns of binge-alcohol use in humans (Eckardt et al., 1998 (link); Jones and Sternebring, 1992 (link)). Further, the relatively moderate ethanol dose employed (i.e., 4g/kg twice daily) allowed for each animal to receive every ethanol dose in the current study; thereby reducing mortality rates known to be inherent to binge-like ethanol administration. Animal weights were recorded prior to 0800 dosing daily.