Each animal received, during three consecutive days; 1) ACTH (1 24) (Synacthen® Depot, Novartis Pharma AG, Basel, Schweiz) at a dose of 10 μg/kg body weight in 5 ml physiological saline, 2) porcine CRH American Peptide Companyy, Inc., Sunnyvale, CA, USA) at a dose 0.6 μg/kg body weight in 5 ml physiological saline or 3) physiological saline (5 ml).
The membrane was washed six times for 5 min in TBST solution before addition of the secondary antibody, HRP labelled goat anti-mouse IgG (Oncogene Research Products, San Diego, CA, USA), at a 1 5000 dilution in 5% (weight:vol) milk/TBST solution for 1 hour at room temperature.
Following MR imaging, a combination of dextran tracers (Texas Red 10 kDa and Fluorescein 500 kDa; Invitrogen Molecular Probes, CA, USA) at a dosage of 5 mg/100 g body weight in sterile saline at a volume of 300-500 μL, was injected into the cardiac left ventricle and allowed to circulate for 60 s, based on previous studies [ 37].
Group 1 received Gem (Biotang, Waltham, MA, USA), at a dose of 50 mg/kg body weight dissolved in saline and DMSO containing PEG-400 (Sigma).
A single intraperitoneal injection of MNU (Ashe Stevens, Detroit, MI, USA) at a dose of 50 mg/kg body weight was given to all rats at 7 weeks of age.
injection of LPS from Escherichia coli (serotype O111 B4, Sigma Chemical Company, St . Louis MO, USA) at a dose of 100 μg/kg of body weight dissolved in 2 mL sterile saline.
After their weight drop, they would plateau at a "weight stability" level.
Weight loss temperatures of the products were measured with a TA thermogravimetric analyzer (TGA 2950, TA Instruments, New Castle, DE, USA) at a heating rate of 20°C/min in N2 atmosphere.
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Weight change of the feed and draw solutions was recorded by an auto logging software (Scort Pro, Ohous, USA) at a time interval of 2 min.
