Liver specimens were evaluated for the degree of steatosis according to Brunt et al., [25], where steatosis was scored as absent (=0), mild when present in <1/3 of the hepatocytes (=1), moderate selleck chemicals llc when present in 1/3�C2/3 of the hepatocytes (=2), and severe when present in >2/3 of the hepatocytes (=3). The presence and location of infiltrating inflammatory cells and liver injury was also recorded. The degree of infiltrating inflammatory cells in steatotic and non-steatotic areas, vascular stasis and loss of liver parenchyma in either zonal or nonzonal distribution were measured using a semi-quantitative graded scale of 0 (absent), 1 (mild), 2 (moderate), and 3 (extensive) [26], to enable statistical evaluation. Bacterial translocation Samples from the caudate lobe of the liver were removed aseptically and frozen immediately at ?70��C until determination.

For analysis, the samples were thawed, placed in an ultrasonic bath (Millipore, Sundbyberg, Sweden) for 5 min and swirled for 2 min on a Chiltern. Viable counts were obtained from Violet-red bile glucose (VRBG) agar (Oxoid) that was incubated aerobically at 37��C for 24 h (Enterobacteriaceae count), brain heart infusion (BHI) agar (Difco, Detroit, MI) that was incubated aerobically and under anaerobic conditions, as described above, at 37��C for 72 h (total aerobic and anaerobic counts, respectively), and from Rogosa agar (Oxoid), incubated anaerobically at 37��C for 72 h (lactobacilli count). Results were expressed as incidences of positive cultures/group.

Colonies were randomly picked from the plates with positive cultures and identified by sequencing the 16 S ribosomal RNA gene. The partial 16 S rRNA gene sequences were searched against GenBank (National Centre for Biotechnology Information, Bethesda, MD) using the Basic Local Alignment Search Tool (BLAST) accessible from the homepage at the National Centre for Biotechnology Information (NCBI; http://www.ncbi.nlm.nih.gov/). The compared sequence lengths were between 200�C900 base pairs. Viable count of Enterobacteriaceae and lactobacilli in faeces Faecal samples were thawed and homogenised in freezing medium, diluted (sodium chloride (Merck), 8.5 g/l; Bacteriological peptone (Oxoid, Unipath LTD Basingstoke, Hampshire, England), 1 g/l; Tween 80 (Merck), 1 g/l; L-Cystine hydrochloride monohydrate (Merck), 0.

2 g/l) and plated on Rogosa agar for lactobacilli count (Oxoid; incubated anaerobically [Gas Pack System, Gas Pack; Becton Dickenson Microbiology Systems, Cockeynsville, MD] at 37��C for 72 h) and violet red bile-glucose agar (VRBG) for Enterobacteriaceae GSK-3 count (Oxoid; incubated aerobically at 37��C for 24 h). The number of colonies formed on each plate was counted and corrected for the weight of the original faecal sample, and expressed as CFU/g faeces.