Cirrhosis severity in patients was previously shown to be associated with progressive changes in the fecal microbiome in a longitudinal setting. Recent evidence shows that bacterial translocation from the gut to the extraintestinal sites could play a major role in poor disease outcome and patient survival. However, the underlying mechanisms involving the microbiota in the disease progression are not well understood. Here, using an animal model of cirrhosis in a longitudinal and multibody sites setting, we showed the presence of a distinct composition of the microbiome in mesenteric lymph nodes, blood, and ascitic fluid compared to that in feces and ileocecal content, suggesting compartmentalization of the gut microbiome. We also demonstrate that microbiome changes in intestinal sites are associated with shifts in specific microbial groups in the mesenteric lymph nodes as well as an increase in systemic cytokine production, linking inflammation to decompensated cirrhosis in the gut-liver-immunity axis.

This study was performed to contribute to the analysis of alpha-lactalbumin "molten globule" state by using spectral and proteolysis techniques. Samples of holo and apo alpha-lactalbumin in the presence of different concentrations of ethanol were analyzed. Results of fluorescence spectroscopy of both forms showed that as ethanol concentration increased, the tryptophanyl residues became more accessible to the solvent. Near circular dichroism spectra of holo alpha-lactalbumin indicated that its tertiary structure was maintained in 20% ethanol whereas it was altered in 30 and 40% ethanol. For apo alpha-lactalbumin, spectra were similar in all samples studied. Holo alpha-lactalbumin was resistant to trypsinolysis in 0% ethanol, whereas it was easily hydrolyzed in 20 and 30% ethanol. In the case of the apo form and in the absence of ethanol, 70% of the protein was degraded after 1 h. However, in the presence of 20 and 30% ethanol, the overall reaction rate was lowered. Peptides obtained after tryptic hydrolysis were identified by reversed-phase high-performance liquid chromatography coupled to mass spectrometry. Differences in population of produced peptides indicate the changes of folding intermediates present in the studied alpha-lactalbumin solutions. This study demonstrated that proteolytic enzymes are suitable tools to determine protein structure complementing physico-chemical studies.

R1033 is a plasmid of compatibility group P (= P1) transferred from a wild strain of Pseudomonas aeruginosa. It confers resistance to gentamicin by gentamicin acetyl-transferase 1 and to kanamycin and neomycin by neomycin phosphotransferase 1.The self-transmissible R factors of Pseudomonas aeruginosa clinical isolates have been assigned to three compatibility groups by Bryan and his collaborators (2)(3)(4). Group P1 contains the R factors that in Escherichia coli K-12 constitute group P (13). Group P2 plasmids are not transmissible to E. coli, and plasmids of group P3 are those assigned to group C (12) or com6 (6) in the E. coli plasmid classifications.Since gentamicin is useful in the treatment of P. aeruginosa infections, attention has been concentrated on mechanisms by which resistance to this drug can be attained. Isolates of P. aeruginosa carrying plasmids conferring gentamicin resistance have been reported (3,17,21 We report here the isolation, in Spain, of a strain resistant to gentamicin (and several other antibiotics) by virtue of an R factor of group P1. MATERIALS AND METHODSBacteria. The bacteria used were P. aeruginosa CRT and E. coli K-12 strains J53-2 (F-, pro,met,rif), J62 (F-, pro,his,trp,lac), and CR34Thy (F-, thr,leu, thi,thy,lac).R factors. The R factors used were Plac, a plasmid of compatibility group C (= P3) which confers lactose fermentation ability and sulfonamide resistance (15), and R751, a plasmid of compatibility group P (= P1) which confers resistance to trimethoprim (18). Phage. The phage used was PRR1, which adsorbs to bacteria carrying plasmids of group P (= P1) and not to plasmid-free becteria or cells carrying plasmids of other groups (11,19). Conjugal plasmid transfer and plasmid compatibility properties. Procedures for conjugal plasmid transfer and the determination of plasmid compatibility properties were as described previously (9, 13).

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