The diversity of interactions, which are among elements of the biological systems, can be studied based on the networks theory. Moreover, the dynamic of these interactions is an inherent trait of those systems. In this sense, several tools have been proposed to compare networks, in that each network represents a state assumed by the system. However, the biological systems generally can assume much more than two biological states and none of the tools are able to compare structural characteristics among more than two networks simultaneously. To solve this issue, we developed a statistical tool to compare two or more networks and highlight key variables of a system. Here we describe the new method, called BioNetStat, that is able to compare correlation networks using traits that are based on graph spectra (the group of eigenvalues of the adjacency matrix), such as the spectral distribution. This measure is associated with several structural characteristics of networks such as the number of walks, diameter, and cliques. In addition to the spectral distribution, BioNetStat can also compare networks to the node centralities. We used two different biological datasets, tumoral cells genes expressions and plant metabolism, to evaluate the performance of BioNetStat and as case studies. The tool is implemented in an R package, and it also has a user-friendly interface. We showed that BioNetStat is efficient in distinguishing more than two networks. In comparison with a similar tool (GSCA), the increase in the number of compared networks reduces less the statistical power of the BioNetStat than the GSCA. Furthermore, BioNetStat is able to find signaling pathways in a bigger proportion than the GSCA, complementing tools proposed in the literature. In the case studies, the method pointed out variables, and sets of variables, with a central role in biological systems, which were not highlighted when only gene expression pattern or metabolomics were studied. For instance, BioNetStat allowed us to differentiate among cancer types and plant organs. The BioNetStat results bring new findings on what differentiate the states, giving us a systemic view of our study subject and affording the proposition of new hypotheses about the studied processes.
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