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Methods: Twenty-five infants were recruited and classified as good and non-/low-responders according to serological test results. Whole genome DNA methylation states were profiled by Illumina HumanMethylation 450 K beadchips. Data were processed through quality and dispersion filtering and with differential methylation analysis based on a combination of average methylation differences and non-parametric statistical tests. Results were finally associated to already published transcriptomics and post-transcriptomics to gain further insight. Background: Variable responses to the Hepatitis B Virus (HBV) vaccine have recently been reported as strongly dependent on genetic causes. Yet, the details on such mechanisms of action are still unknown. In parallel, altered DNA methylation states have been uncovered as important contributors to a variety of health conditions. However, methodologies for the analysis of such high-throughput data (epigenomic), especially from the computational point of view, still lack of a gold standard, mostly due to the intrinsic statistical distribution of methylomic data i.e. binomial rather than (pseudo-) normal, which characterizes the better known transcriptomic data. We present in this article our contribution to the challenge of epigenomic data analysis with application to the variable response to the Hepatitis B virus (HBV) vaccine and its most lethal degeneration: hepatocellular carcinoma (HCC).

BACKGROUND: issues and limitations related to accessibility, understandability and ease of use of signalling pathway databases may hamper or divert research workflow, leading, in the worst case, to the generation of confusing reference frameworks and misinterpretation of experimental results. In an attempt to retrieve signalling pathway data related to a specific set of test genes, we queried and analysed the results from six of the major curated signalling pathway databases: Reactome, PathwayCommons, KEGG, InnateDB, PID, and Wikipathways. FINDINGS: although we expected differences - often a desirable feature for the integration of each individual query, we observed variations of exceptional magnitude, with disproportionate quality and quantity of the results. Some of the more remarkable differences can be explained by the diverse conceptual designs and purposes of the databases, the types of data stored and the structure of the query, as well as by missing or erroneous descriptions of the search procedure. To go beyond the mere enumeration of these problems, we identified a number of operational features, in particular inner and cross coherence, which, once quantified, offer objective criteria to choose the best source of information. CONCLUSIONS: in silico biology heavily relies on the information stored in databases. To ensure that computational biology mirrors biological reality and offers focused hypotheses to be experimentally validated, coherence of data codification is crucial and yet highly underestimated. We make practical recommendations for the end-user to cope with the current state of the databases as well as for the maintainers of those databases to contribute to the goal of the full enactment of the open data paradigm.

The signaling Petri net (SPN) simulator, designed to provide insights into the trends of molecules' activity levels in response to an external stimulus, contributes to the systems biology necessity of analyzing the dynamics of large-scale cellular networks. Implemented into the freely available software, BioLayout Express(3D), the simulator is publicly available and easy to use, provided the input files are prepared in the GraphML format, typically using the network editing software, yEd, and standards specific to the software. However, analysis of complex networks represented using other systems biology formatting languages (on which popular software, such as CellDesigner and Cytoscape, are based) requires manual manipulation, a step that is prone to error and limits the use of the SPN simulator in BioLayout Express(3D). To overcome this, we present a Cytoscape plug-in that enables users to automatically convert networks for analysis with the SPN simulator from the standard systems biology markup language. The automation of this step opens the SPN simulator to a far larger user group than has previously been possible.

Microarray and deep sequencing technologies have provided unprecedented opportunities for mapping genome mutations, RNA transcripts, transcription factor binding, and histone modifications at high resolution at the genome-wide level. This has revolutionized the way in which transcriptomes, regulatory networks and epigenetic regulations have been studied and large amounts of heterogeneous data have been generated. Although efforts are being made to integrate these datasets unbiasedly and efficiently, how best to do this still remains a challenge. Here we review major impacts of high-throughput genome-wide data generation, their relevance to human diseases, and various bioinformatics approaches for data integration. Finally, we provide a case study on inflammatory diseases.

Results: Analysis of the CRKL network -available at http://www.picb.ac.cn/ClinicalGenomicNTW/software.html-allows for investigation of the potential effect of perturbing genes of interest. Within the group of genes that are significantly affected by simulated perturbation of CRKL, we are lead to further investigate the importance of PXN. Our results allow us to (1) refine the hypothesis on CRKL as a novel drug target (2) indicate potential causes of side effects in on- going trials and (3) importantly, provide recommendations with impact on on- going clinical studies. Background: Rheumatoid arthritis (RA) is among the most common human systemic autoimmune diseases, affecting approximately 1% of the population worldwide. To date, there is no cure for the disease and current treatments show undesirable side effects. As the disease affects a growing number of individuals, and during their working age, the gathering of all information able to improve therapies -by understanding their and the disease mechanisms of action- represents an important area of research, benefiting not only patients but also societies. In this direction, network analysis methods have been used in previous work to further our understanding of this complex disease, leading to the identification of CRKL as a potential drug target for treatment of RA. Here, we use computational methods to expand on this work, testing the hypothesis in silico.

Results: Our approach MIMO (Molecular Interaction Maps Overlap) addresses the first problem by allowing the introduction of gaps and mismatches between query and template pathways and permits - when necessary-supervised queries incorporating a priori biological information. It then addresses the second issue by relying directly on the rich graph topology described in the Systems Biology Markup Language (SBML) standard, and uses multidigraphs to efficiently handle multiple queries on biological graph databases. The algorithm has been here successfully used to highlight the contact point between various human pathways in the Reactome database. Background: Molecular pathways represent an ensemble of interactions occurring among molecules within the cell and between cells. The identification of similarities between molecular pathways across organisms and functions has a critical role in understanding complex biological processes. For the inference of such novel information, the comparison of molecular pathways requires to account for imperfect matches (flexibility) and to efficiently handle complex network topologies. To date, these characteristics are only partially available in tools designed to compare molecular interaction maps.

Results: We address the latter of these two challenges by testing an integrated approach on a known cancer benchmark: the NCI-60 cell panel. Here, high-throughput screens for mRNA, miRNA and proteins are jointly analyzed using factor analysis, combined with linear discriminant analysis, to identify the molecular characteristics of cancer. Comparisons with separate (non-joint) analyses show that the proposed integrated approach can uncover deeper and more precise biological information. In particular, the integrated approach gives a more complete picture of the set of miRNAs identified and the Wnt pathway, which represents an important surrogate marker of melanoma progression. We further test the approach on a more challenging patient-dataset, for which we are able to identify clinically relevant markers. Background: High-throughput (omic) data have become more widespread in both quantity and frequency of use, thanks to technological advances, lower costs and higher precision. Consequently, computational scientists are confronted by two parallel challenges: on one side, the design of efficient methods to interpret each of these data in their own right (gene expression signatures, protein markers, etc.) and, on the other side, realization of a novel, pressing request from the biological field to design methodologies that allow for these data to be interpreted as a whole, i.e. not only as the union of relevant molecules in each of these layers, but as a complex molecular signature containing proteins, mRNAs and miRNAs, all of which must be directly associated in the results of analyses that are able to capture inter-layers connections and complexity.

Advances in experimental biology, coupled with advances in computational power, bring new challenges to the interdisciplinary field of computational biology. One such broad challenge lies in the reverse engineering of gene networks, and goes from determining the structure of static networks, to reconstructing the dynamics of interactions from time series data. Here, we focus our attention on the latter area, and in particular, on parameterizing a dynamic network of oriented interactions between genes. By basing the parameterizing approach on a known power-law relationship model between connected genes (S-system), we are able to account for non-linearity in the network, without compromising the ability to analyze network characteristics. In this article, we introduce the S-System Parameter Estimation Method (SPEM). SPEM, a freely available R software package (http://www.picb.ac.cn/ClinicalGenomicNTW/temp3.html), takes gene expression data in time series and returns the network of interactions as a set of differential equations. The methods, which are presented and tested here, are shown to provide accurate results not only on synthetic data, but more importantly on real and therefore noisy by nature, biological data. In summary, SPEM shows high sensitivity and positive predicted values, as well as free availability and expansibility (because based on open source software). We expect these characteristics to make it a useful and broadly applicable software in the challenging reconstruction of dynamic gene networks.

Translational and evidence based medicine can take advantage of biotechnology advances that offer a fast growing variety of high-throughput data for screening molecular activities of genomic, transcriptional, post-transcriptional and translational observations. The clinical information hidden in these data can be clarified with clinical bioinformatics approaches. We have recently proposed a method to analyze different layers of high-throughput (omic) data to preserve the emergent properties that appear in the cellular system when all molecular levels are interacting. We show here that this method applied to brain cancer data can uncover properties (i.e. molecules related to protective versus risky features in different types of brain cancers) that have been independently validated as survival markers, with potential important application in clinical practice. © 2012 Fronza et al.; licensee BioMed Central Ltd.

Regenerative medicine is a novel clinical branch aiming at the cure of diseases by replacement of damaged tissues. The crucial use of stem cells makes this area rich of challenges, given the poorly understood mechanisms of differentiation. One highly needed and yet unavailable technology should allow us to monitor the exact (metabolic) state of stem cells differentiation to maximize the effectiveness of their implant in vivo. This is challenged by the fact that not all relevant metabolites in stem cells differentiation are known and not all metabolites can currently be continuously monitored. To bring advancements in this direction, we propose the enhancement and integration of two available technologies into a general pipeline. Namely, high-throughput biochip for gene expression screening to pre-select the variables that are most likely to be relevant in the identification of the stem cells' state and low-throughput biochip for continuous monitoring of cell metabolism with highly sensitive carbon nanotubes-based sensors. Intriguingly, additionally to the involvement of multidisciplinary expertise (medicine, molecular biology, computer science, engineering, and physics), this whole query heavily relies on biochips: it starts in fact from the use of high-throughput ones, which output, in turn, becomes the base for the design of low-throughput, highly sensitive biochips. Future research is warranted in this direction to develop and validated the proposed device.

Community structure is an important topological phenomenon typical of complex networks. Accurately unveiling communities is thus crucial to understand and capture the many-faceted nature of complex networks. Communities in real world frequently overlap, i.e. nodes can belong to more than one community. Therefore, quantitatively evaluating the extent to which a node belongs to a community is a key step to find overlapping boundaries between communities. Non-negative matrix factorization (NMF) is a technique that has been used to detect overlapping communities. However, previous efforts in this direction present: (i) limitations in the interpretation of meaningful overlaps and (ii) lack of accuracy in predicting the correct number of communities. In this paper, a hybrid method of NMF to overcome both limitations is presented. This approach effectively estimates the number of communities and is more interpretable and more accurate in identifying overlapping communities in undirected networks than previous approaches. Validations on synthetic and real world networks show that the proposed community learning framework can effectively reveal overlapping communities in complex networks.

Results: We use Factor Analysis coupled with pre-established knowledge as a theoretical base to achieve this goal. Our intention is to identify structures that contain information from both mRNAs and miRNAs, and that can explain the complexity of the data. Despite the small sample available, we can show that this approach permits identification of meaningful structures, in particular two polycistronic miRNA genes related to transcriptional activity and likely to be relevant in the discrimination between gliosarcomas and other brain tumors. Background: Advances in biotechnology offer a fast growing variety of high-throughput data for screening molecular activities of genomic, transcriptional, post-transcriptional and translational observations. However, to date, most computational and algorithmic efforts have been directed at mining data from each of these molecular levels (genomic, transcriptional, etc.) separately. In view of the rapid advances in technology (new generation sequencing, high-throughput proteomics) it is important to address the problem of analyzing these data as a whole, i.e. preserving the emergent properties that appear in the cellular system when all molecular levels are interacting. We analyzed one of the (currently) few datasets that provide both transcriptional and post-transcriptional data of the same samples to investigate the possibility to extract more information, using a joint analysis approach.

Community structures are found to exist ubiquitously in a number of systems conveniently represented as complex networks. Partitioning networks into communities is thus important and crucial to both capture and simplify these systems' complexity. The prevalent and standard approach to meet this goal is related to the maximization of a quality function, modularity, which measures the goodness of a partition of a network into communities. However, it has recently been found that modularity maximization suffers from a resolution limit, which prevents its effectiveness and range of applications. Even when neglecting the resolution limit, methods designed for detecting communities in undirected networks cannot always be easily extended, and even less directly applied, to directed networks (for which specifically designed community detection methods are very limited). Furthermore, real-world networks are frequently found to possess hierarchical structure and the problem of revealing such type of structure is far from being addressed. In this paper, we propose a scheme that partitions networks into communities by electing community leaders via message passing between nodes. Using random walk on networks, this scheme derives an effective similarity measure between nodes, which is closely related to community memberships of nodes. Importantly, this approach can be applied to a very broad range of networks types. In fact, the successful validation of the proposed scheme on real and synthetic networks shows that this approach can effectively (i) address the problem of resolution limit and (ii) find communities in both directed and undirected networks within a unified framework, including revealing multiple levels of robust community partitions.

Results: The algorithm was first validated on synthetic and real benchmarks. It was then applied to the reconstruction of the core of the amino acids metabolism in Bifidobacterium longum, an essential, yet poorly known player in the human gut intestinal microbiome, known to be related to the onset of important diseases, such as metabolic syndromes. Our results show how computational approaches can offer effective tools for applications with the identification of potential new biological information. Motivation: The reliable and reproducible identification of gene interaction networks represents one of the grand challenges of both modern molecular biology and computational sciences. Approaches based on careful collection of literature data and network topological analysis, applied to unicellular organisms, have proven to offer results applicable to medical therapies. However, when little a priori knowledge is available, other approaches, not relying so strongly on previous literature, must be used. We propose here a novel algorithm ( based on ordinary differential equations) able to infer the interactions occurring among genes, starting from gene expression steady state data.

This new book examines the latest research in the synthetic biology which refers to both: the design and fabrication of biological components and systems that do not already exist in the natural world the re-design and fabrication of existing biological systems. It also deals with Integrative biology is the study and research of biological systems. It does not simply involve one discipline, but integrates a wide variety of disciplines that work together to find answers to scientific questions.

SBML model of rheumatoid arthritis

Background: Computational biology contributes to a variety of areas related to life sciences and, due to the growing impact of translational medicine - the scientific approach to medicine in tight relation with basic science, it is becoming an important player in clinical-related areas. In this study, we use computation methods in order to improve our understanding of the complex interactions that occur between molecules related to Rheumatoid Arthritis (RA). Methodology: Due to the complexity of the disease and the numerous molecular players involved, we devised a method to construct a systemic network of interactions of the processes ongoing in patients affected by RA. The network is based on high-throughput data, refined semi-automatically with carefully curated literature-based information. This global network has then been topologically analysed, as a whole and tissue-specifically, in order to translate the experimental molecular connections into topological motifs meaningful in the identification of tissue-specific markers and targets in the diagnosis, and possibly in the therapy, of RA. Significance: We find that some nodes in the network that prove to be topologically important, in particular AKT2, IL6, MAPK1 and TP53, are also known to be associated with drugs used for the treatment of RA. Importantly, based on topological consideration, we are also able to suggest CRKL as a novel potentially relevant molecule for the diagnosis or treatment of RA. This type of finding proves the potential of in silico analyses able to produce highly refined hypotheses, based on vast experimental data, to be tested further and more efficiently. As research on RA is ongoing, the present map is in fieri, despite being -at the moment- a reflection of the state of the art. For this reason we make the network freely available in the standardised and easily exportable.xml CellDesigner format at 'www.picb.ac.cn/ClinicalGenomicNTW/temp.html' and 'www.celldesigner.org'. © 2010 Wu et al.

Community structures are found to exist ubiquitously in real-world complex networks. We address here the problem of community detection in directed networks. Most of the previous literature ignores edge directions and applies methods designed for community detection in undirected networks, which discards valuable information and often fails when different communities are defined on the basis of incoming and outgoing edges. We suggest extracting information about edge directions using a PageRank random walk and translating such information into edge weights. After extraction we obtain a new weighted directed network in which edge directions can then be safely ignored. We thus transform community detection in directed networks into community detection in reweighted undirected networks. Such an approach can benefit directly from the large volume of algorithms for the detection of communities in undirected networks already developed, since it is not obvious how to extend these algorithms to account for directed networks and the procedure is often difficult. Validations on synthetic and real-world networks demonstrate that the proposed framework can effectively detect communities in directed networks.

Understanding the complexity of the cellular machinery represents a grand challenge in molecular biology. To contribute to the deconvolution of this complexity, a novel inference algorithm based on linear ordinary differential equations is proposed, based solely on high-throughput gene expression data. The algorithm can infer (i) gene-gene interactions from steady state expression profiles and (ii) mode-of-action of the components that can trigger changes in the system. Results demonstrate that the proposed algorithm can identify both information with high performances, thus overcoming the limitation of current algorithms that can infer reliably only one.