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imal. Detection of animal location and movements is performed with infrared sensor pairs arranged in strips for horizontal activity, discriminating in between ambulatory and fine movements. The LabMaster also consists of a combination of very sensitive feeding and drinking sensors for automated on the web measurements. Before recording, animals had been permitted a -day acclimatization period in instruction cages. Measurements have been performed more than the h of saline or dexamethasone infusion. Controls Icv treated Wistar rats Pre-treatment food intake Post-treatment physique weight Glucose NEFA TG Icv treated SD rats Pre-treatment meals intake Post-treatment physique weight Glucose NEFA TG Lipid WAT Liver TG Sc treated SD rats Glucose NEFA TG .. Dex mgday .. Physique composition An EchoMRI- quantitative nuclear magnetic resonance analyzer was utilized to measure total fat and lean body mass in the finish in the therapies. Also, various white adipose tissue depots have been cautiously dissected and weighted immediately after sacrifice. Values represent imply SEM of n = rats per group. P,. in comparison to manage rats, working with Student's t test. doi:.journal.pone..t Plasma measurements Plasma glucose was measured by the glucose oxidase technique. Plasma nonesterified fatty acids and triglycerides were determined making use of Wako Chemicals GmbH and bioMerieux commercial kits, respectively. Plasma corticosterone and leptin levels were determined employing a double antibody RIA having a intra-assay coefficient of variation, and plasma insulin levels have been measured by RIA, as previously described. In one of several experiments, plasma corticosterone and -dehydrocorticosterone were also measured by LC-MSMS. Plasma dexamethasone levels following icv treatment in Wistar rats have been measured by mass spectrometry. Briefly, dexamethasone was quantified by liquid chromatography tandem mass spectrometry employing as Surveyor HPLC, equipped having a Allure biphenyl column linked to a TSQ Quantum mass spectrometer operated in electrospray mode as reported ahead of. Steroids had been eluted employing a mobile phase of :, . mlmin, uC) and quantified versus a calibration curve. Mass transitions monitored have been dexamethasone, D-dexamethasone. Parke-Davis and Bayer, Leverkusen, Switzerland) at and mg kg, respectively, and implanted using a cannula within the suitable lateral cerebral ventricle. Following one week of recovery, the drinking response to icv injection of Angiotensin II was measured to confirm the appropriate placement of your cannula. Osmotic minipumps delivering either the vehicle or dexamethasone diluted in . NaCl have been then subcutaneously implanted and connected to the icv cannula through a polyethylene catheter, as previously described. From results obtained on the physique weight obtain, immediately after a pilot dose-response study, a dose of mgday was Pomalidomide Fda Guidance lastly selected, as previously published. Wistar rats had been treated for days and SD rats for days to obtain equivalent physique weight gains. Animals were then sacrificed inside the fed state, utilizing isoflurane anesthesia and fast decapitation involving and h. Blood samples have been collected into EDTA-coated tubes, centrifuged and plasma stored at uC till use. Tissues were quickly removed, freeze-clamped, and stored at uC. Peripheral infusion of dexamethasone Soon after one week of adaptation, SD rats were individually housed and submitted to surgical procedures. They have been briefly anesthetized with isoflurane and osmotic minipumps delivering either the automobile or mgday dexamethasone were subcutaneously implanted for two days. Rats