Diet remained constant throughout the study and there was no change in the mead acid level, which is a marker for dietary intake change between the pre and post-rifaximin profile [40]. Therefore the increased fatty acids are likely either due to an enhanced transport from the gut to the bloodstream via the thoracic duct as chylomicrons or enhanced release from the adipose tissue. Gut microbiota can affect adipose tissue and peripheral lipoprotein lipase by modulating the fasting-induced adipose P7C3 site factor [41,42]. The lack of short-chain fatty acids, which are major end-products of bacterial fermentation, in this serum profile is likely because the majority of their biological activity occurs within the gut lumen and they are directly absorbed and transported into the liver [43]. The predominance of long-chain serum fatty acids in the postrifaximin profile supports the gut-based transport of these molecules in chylomicrons as a potential mechanism for their higher levels. Prior studies have shown that fatty acids, both saturated and unsaturated, are associated with brain function in animal, human and population-based studies[39,44?6]. The brain fatty acid profile impacts neurogenesis, cognition and memory possibly by affecting neurotransmission, axonal sheath composition and cell membrane fluidity. Fatty acids increased in our study, arachidonic and linoleic acids, have been shown to influence brain function directly [44,46]. There was a significantTable 2. Comparison of Thiazole Orange network topology before and after rifaximin.Before Rifaximin Number of Nodes Isolated Nodes Connected Components Average Number of Neighbors Network Density Clustering Coefficient (saturation of the nodes) Network Diameter (largest distance between nodes) Network Radius (shortest distance between nodes) Characteristic Path Length (expected distance between two nodes) Network Centralization Shortest Path (shortest path through all nodes) Network Heterogeneity (tendency to form hubs) 2220 0 1 59.0405405 0.02660682 0.36257932 6 4 2.77271111 0.23453281 4926180 1.After Rifaximin 2225 0 1 51.4588764 0.02313798 0.33746817 6 4 2.75946771 0.18386182 4948400 1.Intersection of the two networks 2219 511 547 13.5205047 0.00609581 0.31452636 15 1 4.68771603 0.15184087 2600364 1.Intersection indicates the nodes and network common to both before and after rifaximin. The table shows that the majority of nodes involved were common (intersection) between the groups while the network density (average number of neighbors and network density) changed after rifaximin therapy. While the diameter and radius remained same, there was a reduction in the path length and heterogeneity after rifaximin compared to before. There was also a decrease in network centralization which means that the distribution was spread out after rifaximin therapy compared to before. doi:10.1371/journal.pone.0060042.tMetabiome and Rifaximin in CirrhosisFigure 5. Subset of correlation differences before and after rifaximin. This figure is limited to the metabolomics and clinical/cognitive features that changed with rifaximin and their interaction with the bacterial taxa. The linkages that significantly changed in nature (positive to negative or vice-versa) or intensity (less to more or vice-versa while remaining positive or negative) with p,0.05 are shown. Nodes: Blue: bacterial taxa, green: serum metabolites, Yellow: cognitive or clinical data. Linkages were dark blue if correlations were positive before and changed signific.Diet remained constant throughout the study and there was no change in the mead acid level, which is a marker for dietary intake change between the pre and post-rifaximin profile [40]. Therefore the increased fatty acids are likely either due to an enhanced transport from the gut to the bloodstream via the thoracic duct as chylomicrons or enhanced release from the adipose tissue. Gut microbiota can affect adipose tissue and peripheral lipoprotein lipase by modulating the fasting-induced adipose factor [41,42]. The lack of short-chain fatty acids, which are major end-products of bacterial fermentation, in this serum profile is likely because the majority of their biological activity occurs within the gut lumen and they are directly absorbed and transported into the liver [43]. The predominance of long-chain serum fatty acids in the postrifaximin profile supports the gut-based transport of these molecules in chylomicrons as a potential mechanism for their higher levels. Prior studies have shown that fatty acids, both saturated and unsaturated, are associated with brain function in animal, human and population-based studies[39,44?6]. The brain fatty acid profile impacts neurogenesis, cognition and memory possibly by affecting neurotransmission, axonal sheath composition and cell membrane fluidity. Fatty acids increased in our study, arachidonic and linoleic acids, have been shown to influence brain function directly [44,46]. There was a significantTable 2. Comparison of network topology before and after rifaximin.Before Rifaximin Number of Nodes Isolated Nodes Connected Components Average Number of Neighbors Network Density Clustering Coefficient (saturation of the nodes) Network Diameter (largest distance between nodes) Network Radius (shortest distance between nodes) Characteristic Path Length (expected distance between two nodes) Network Centralization Shortest Path (shortest path through all nodes) Network Heterogeneity (tendency to form hubs) 2220 0 1 59.0405405 0.02660682 0.36257932 6 4 2.77271111 0.23453281 4926180 1.After Rifaximin 2225 0 1 51.4588764 0.02313798 0.33746817 6 4 2.75946771 0.18386182 4948400 1.Intersection of the two networks 2219 511 547 13.5205047 0.00609581 0.31452636 15 1 4.68771603 0.15184087 2600364 1.Intersection indicates the nodes and network common to both before and after rifaximin. The table shows that the majority of nodes involved were common (intersection) between the groups while the network density (average number of neighbors and network density) changed after rifaximin therapy. While the diameter and radius remained same, there was a reduction in the path length and heterogeneity after rifaximin compared to before. There was also a decrease in network centralization which means that the distribution was spread out after rifaximin therapy compared to before. doi:10.1371/journal.pone.0060042.tMetabiome and Rifaximin in CirrhosisFigure 5. Subset of correlation differences before and after rifaximin. This figure is limited to the metabolomics and clinical/cognitive features that changed with rifaximin and their interaction with the bacterial taxa. The linkages that significantly changed in nature (positive to negative or vice-versa) or intensity (less to more or vice-versa while remaining positive or negative) with p,0.05 are shown. Nodes: Blue: bacterial taxa, green: serum metabolites, Yellow: cognitive or clinical data. Linkages were dark blue if correlations were positive before and changed signific.