Modern scientific discovery is frequently backed by largescale scientific experiments, which cannot prescind from the unparalleled computational capabilities provided by highperformance computing systems. However, large data centers usually prioritize system efficiency over user accessibility, posing challenges for researchers without advanced computer science expertise. This work introduces BookedSlurm, a secure and userfocused extension of the Slurm workload manager, aiming to democratize HPC access across interdisciplinary research domains. BookedSlurm enables a partially decentralized regulation of finegrained advanced resource reservations through a novel creditbased framework, ensuring fair and predictable access to computing resources. Its modular architecture leverages dedicated microservices to manage reservations, credit handling, and accounting. These components are exposed through a secure REST API and an intuitive webbased dashboard, enhancing system usability for novice and expert users. While the dashboard simplifies interactions for nonspecialists, advanced users can directly access agentlevel APIs for more complex and automated operations. The effectiveness of BookedSlurm is validated on a realworld bioinformatics use case from the SUSMIRRI.IT project, showcasing its ability to enhance usability, optimize job scheduling, and streamline execution workflows.

MOTIVATION: Computational models are crucial for addressing critical questions about systems evolution and deciphering system connections. The pivotal feature of making this concept recognizable from the biological and clinical community is the possibility of quickly inspecting the whole system, bearing in mind the different granularity levels of its components. This holistic view of system behaviour expands the evolution study by identifying the heterogeneous behaviours applicable, e.g. to the cancer evolution study. RESULTS: To address this aspect, we propose a new modelling paradigm, UnifiedGreatMod, which allows modellers to integrate fine-grained and coarse-grained biological information into a unique model. It enables functional studies by combining the analysis of the system's multi-level stable states with its fluctuating conditions. This approach helps to investigate the functional relationships and dependencies among biological entities. This is achieved, thanks to the hybridization of two analysis approaches that capture a system's different granularity levels. The proposed paradigm was then implemented into the open-source, general modelling framework GreatMod, in which a graphical meta-formalism is exploited to simplify the model creation phase and R languages to define user-defined analysis workflows. The proposal's effectiveness was demonstrated by mechanistically simulating the metabolic output of Escherichia coli under environmental nutrient perturbations and integrating a gene expression dataset. Additionally, the UnifiedGreatMod was used to examine the responses of luminal epithelial cells to Clostridium difficile infection. AVAILABILITY AND IMPLEMENTATION: GreatMod https://qbioturin.github.io/epimod/, epimod_FBAfunctions https://github.com/qBioTurin/epimod_FBAfunctions, first case study E. coli https://github.com/qBioTurin/Ec_coli_modelling, second case study C. difficile https://github.com/qBioTurin/EpiCell_CDifficile.

BACKGROUND: The analysis of large and complex biological datasets in bioinformatics poses a significant challenge to achieving reproducible research outcomes due to inconsistencies and the lack of standardization in the analysis process. These issues can lead to discrepancies in results, undermining the credibility and impact of bioinformatics research and creating mistrust in the scientific process. To address these challenges, open science practices such as sharing data, code, and methods have been encouraged. RESULTS: CREDO, a Customizable, REproducible, DOcker file generator for bioinformatics applications, has been developed as a tool to moderate reproducibility issues by building and distributing docker containers with embedded bioinformatics tools. CREDO simplifies the process of generating Docker images, facilitating reproducibility and efficient research in bioinformatics. The crucial step in generating a Docker image is creating the Dockerfile, which requires incorporating heterogeneous packages and environments such as Bioconductor and Conda. CREDO stores all required package information and dependencies in a Github-compatible format to enhance Docker image reproducibility, allowing easy image creation from scratch. The user-friendly GUI and CREDO's ability to generate modular Docker images make it an ideal tool for life scientists to efficiently create Docker images. Overall, CREDO is a valuable tool for addressing reproducibility issues in bioinformatics research and promoting open science practices.

Open science and reproducibility are two key pillars of modern scientific research. Open science is making scientific research and data accessible and transparent to the broader scientific community and the public. Reproducibility, on the other hand, is the ability to replicate and confirm research results by following the same methods and procedures. Reproducibility is thus crucial because it ensures the reliability and validity of scientific findings. The relationship between open science and reproducibility is intertwined; indeed open science practices, such as sharing raw data, detailed methodologies, and code, greatly facilitate the reproducibility of research. In recent years, concerns about the reproducibility of scientific research have gained prominence, and indeed scientists still lament the lack of details in the methods sections of published papers and the unavailability of raw data from the authors.To assist cellular biologists and immunologists and to promote a more transparent, open and reproducible research practice, we developed OmniReproducibleCellAnalysis (ORCA), a new Shiny Application based in R, for the semi-automated analysis of Western Blot (WB), Reverse Transcription-quantitative PCR (RT-qPCR), Enzyme-Linked ImmunoSorbent Assay (ELISA), Endocytosis and Cytotoxicity experiments. ORCA is open-source and approachable by scientists without advanced R language knowledge. Our application automatically compiles a report containing the finalized data analysis and all its preliminary and intermediate steps, ensuring data analysis standardization and reproducibility. Furthermore, ORCA allows to upload raw data and results directly on the data repository Harvard Dataverse, a valuable tool for promoting transparency and data accessibility in scientific research.By employing ORCA, scientists will cut down analysis time and human-dependent errors, while taking a step towards a research practice compliant with Open Science and FAIR principle.

Multi-omics analysis aims at extracting previously uncovered biological knowledge by integrating information across multiple single-omic sources. Past approaches have focused on the simultaneous analysis of a small number of omic data sets. Current challenges face the problem of integrating multiple omic sources into a unified complex model, or of combining already available tools for two-by-two omics analyses and merging their outcomes. By doing so and leveraging integrated system-level knowledge, multi-omic approaches ought to enable the development of better qualitative and quantitative models for descriptive and predictive analyses. To move this area forward, new statistical and algorithmic frameworks are needed, for example for generalizing classical graph theory results to heterogeneous networks, and applying them to diverse problems such as drug repurposing or understanding the immune response to infections. Thus, in short, this workshop aims at investigating novel methodologies for providing crucial insights into multi-omics data management, integration, and analysis to enable biological discoveries. The workshop will be sponsored by the InfoLife CINI National Laboratory (https://www.consorzio-cini.it/index.php/en/ ).

MOTIVATION: The transition from evaluating a single time point to examining the entire dynamic evolution of a system is possible only in the presence of the proper framework. The strong variability of dynamic evolution makes the definition of an explanatory procedure for data fitting and clustering challenging. RESULTS: We developed CONNECTOR, a data-driven framework able to analyze and inspect longitudinal data in a straightforward and revealing way. When used to analyze tumor growth kinetics over time in 1599 patient-derived xenograft growth curves from ovarian and colorectal cancers, CONNECTOR allowed the aggregation of time-series data through an unsupervised approach in informative clusters. We give a new perspective of mechanism interpretation, specifically, we define novel model aggregations and we identify unanticipated molecular associations with response to clinically approved therapies. AVAILABILITY AND IMPLEMENTATION: CONNECTOR is freely available under GNU GPL license at https://qbioturin.github.io/connector and https://doi.org/10.17504/protocols.io.8epv56e74g1b/v1.

OBJECTIVE: Computational models are at the forefront of the pursuit of personalized medicine thanks to their descriptive and predictive abilities. In the presence of complex and heterogeneous data, patient stratification is a prerequisite for effective precision medicine, since disease development is often driven by individual variability and unpredictable environmental events. Herein, we present GreatNectorworkflow as a valuable tool for (i) the analysis and clustering of patient-derived longitudinal data, and (ii) the simulation of the resulting model of patient-specific disease dynamics. METHODS: GreatNectoris designed by combining an analytic strategy composed of CONNECTOR, a data-driven framework for the inspection of longitudinal data, and an unsupervised methodology to stratify the subjects with GreatMod, a quantitative modeling framework based on the Petri Net formalism and its generalizations. RESULTS: To illustrate GreatNectorcapabilities, we exploited longitudinal data of four immune cell populations collected from Multiple Sclerosis patients. Our main results report that the T-cell dynamics after alemtuzumab treatment separate non-responders versus responders patients, and the patients in the non-responders group are characterized by an increase of the Th17 concentration around 36 months. CONCLUSION: GreatNectoranalysis was able to stratify individual patients into three model meta-patients whose dynamics suggested insight into patient-tailored interventions.

The immune system (IS) represents a complex network of cells and molecules devoted to the protection of individuals from external pathogens, and in terms of complexity, it is only second to the central nervous system. As our knowledge of the IS mechanisms has become more exhaustive, interest has grown in applying modeling and simulation techniques in this context. In particular, among these techniques, the Agent Based Models (ABMs) have been increasingly applied for the IS simulation. One of the major drawbacks of ABMs is represented by the lack of well-defined semantics, which may lead to inconsistent results in comparison to other stochastic approaches. In this paper, we make use of the well-defined semantics and the simulation algorithm for ABMs that we proposed in [1] to implement a few models of the Cancer-Immune System. Comparing ABMs and Gillespie’s Stochastic Simulation Algorithm results we show that our methodology brings coherence among the results of ABMs and SSA.

Multi-omics analysis aims at extracting previously uncovered biological knowledge by integrating information across multiple single-omic sources. Past approaches have focused on the simultaneous analysis of a small number of omic data sets. Current challenges face the problem of integrating multiple omic sources into a unified complex model, or of combining already available tools for two-by-two omics analyses and merging their outcomes. By doing so and leveraging integrated system-level knowledge, multi-omic approaches ought to enable the development of better qualitative and quantitative models for descriptive and predictive analyses. To move this area forward, new statistical and algorithmic frameworks are needed, for example for generalizing classical graph theory results to heterogeneous networks and applying them to diverse problems such as drug repurposing or understanding the immune response to infections. Thus, in short, this workshop aims at investigating novel methodologies for providing crucial insights into multi-omics data management, integration, and analysis in order to enable biological discoveries.

BACKGROUND: Graphs are mathematical structures widely used for expressing relationships among elements when representing biomedical and biological information. On top of these representations, several analyses are performed. A common task is the search of one substructure within one graph, called target. The problem is referred to as one-to-one subgraph search, and it is known to be NP-complete. Heuristics and indexing techniques can be applied to facilitate the search. Indexing techniques are also exploited in the context of searching in a collection of target graphs, referred to as one-to-many subgraph problem. Filter-and-verification methods that use indexing approaches provide a fast pruning of target graphs or parts of them that do not contain the query. The expensive verification phase is then performed only on the subset of promising targets. Indexing strategies extract graph features at a sufficient granularity level for performing a powerful filtering step. Features are memorized in data structures allowing an efficient access. Indexing size, querying time and filtering power are key points for the development of efficient subgraph searching solutions. RESULTS: An existing approach, GRAPES, has been shown to have good performance in terms of speed-up for both one-to-one and one-to-many cases. However, it suffers in the size of the built index. For this reason, we propose GRAPES-DD, a modified version of GRAPES in which the indexing structure has been replaced with a Decision Diagram. Decision Diagrams are a broad class of data structures widely used to encode and manipulate functions efficiently. Experiments on biomedical structures and synthetic graphs have confirmed our expectation showing that GRAPES-DD has substantially reduced the memory utilization compared to GRAPES without worsening the searching time. CONCLUSION: The use of Decision Diagrams for searching in biochemical and biological graphs is completely new and potentially promising thanks to their ability to encode compactly sets by exploiting their structure and regularity, and to manipulate entire sets of elements at once, instead of exploring each single element explicitly. Search strategies based on Decision Diagram makes the indexing for biochemical graphs, and not only, more affordable allowing us to potentially deal with huge and ever growing collections of biochemical and biological structures.

BACKGROUND: Emerging and re-emerging infectious diseases such as Zika, SARS, ncovid19 and Pertussis, pose a compelling challenge for epidemiologists due to their significant impact on global public health. In this context, computational models and computer simulations are one of the available research tools that epidemiologists can exploit to better understand the spreading characteristics of these diseases and to decide on vaccination policies, human interaction controls, and other social measures to counter, mitigate or simply delay the spread of the infectious diseases. Nevertheless, the construction of mathematical models for these diseases and their solutions remain a challenging tasks due to the fact that little effort has been devoted to the definition of a general framework easily accessible even by researchers without advanced modelling and mathematical skills. RESULTS: In this paper we describe a new general modeling framework to study epidemiological systems, whose novelties and strengths are: (1) the use of a graphical formalism to simplify the model creation phase; (2) the implementation of an R package providing a friendly interface to access the analysis techniques implemented in the framework; (3) a high level of portability and reproducibility granted by the containerization of all analysis techniques implemented in the framework; (4) a well-defined schema and related infrastructure to allow users to easily integrate their own analysis workflow in the framework. Then, the effectiveness of this framework is showed through a case of study in which we investigate the pertussis epidemiology in Italy. CONCLUSIONS: We propose a new general modeling framework for the analysis of epidemiological systems, which exploits Petri Net graphical formalism, R environment, and Docker containerization to derive a tool easily accessible by any researcher even without advanced mathematical and computational skills. Moreover, the framework was implemented following the guidelines defined by Reproducible Bioinformatics Project so it guarantees reproducible analysis and makes simple the developed of new user-defined workflows.

BACKGROUND: Multiple Sclerosis (MS) represents nowadays in Europe the leading cause of non-traumatic disabilities in young adults, with more than 700,000 EU cases. Although huge strides have been made over the years, MS etiology remains partially unknown. Furthermore, the presence of various endogenous and exogenous factors can greatly influence the immune response of different individuals, making it difficult to study and understand the disease. This becomes more evident in a personalized-fashion when medical doctors have to choose the best therapy for patient well-being. In this optics, the use of stochastic models, capable of taking into consideration all the fluctuations due to unknown factors and individual variability, is highly advisable. RESULTS: We propose a new model to study the immune response in relapsing remitting MS (RRMS), the most common form of MS that is characterized by alternate episodes of symptom exacerbation (relapses) with periods of disease stability (remission). In this new model, both the peripheral lymph node/blood vessel and the central nervous system are explicitly represented. The model was created and analysed using Epimod, our recently developed general framework for modeling complex biological systems. Then the effectiveness of our model was shown by modeling the complex immunological mechanisms characterizing RRMS during its course and under the DAC administration. CONCLUSIONS: Simulation results have proven the ability of the model to reproduce in silico the immune T cell balance characterizing RRMS course and the DAC effects. Furthermore, they confirmed the importance of a timely intervention on the disease course.

Computational Biology is a fast-growing field that is enriched by different data-driven methodological approaches and by findings and applications in a broad range of biological areas. Fundamental to these approaches are the mathematical and computational models used to describe the different state s at microscopic (for example a biochemical reaction), mesoscopic (the signalling effects at tissue level), and macroscopic levels (physiological and pathological effects) of biological processes. In this paper we address the problem of combining two powerful classes of methodologies: Flux Balance Analysis (FBA) methods which are now producing a revolution in biotechnology and medicine, and Petri Nets (PNs) which allow system generalisation and are central to various mathematical treatments, for example Ordinary Differential Equation (ODE) specification of the biosystem under study. While the former is limited to modelling metabolic networks, i.e. does not account for intermittent dynamical signalling events, the latter is hampered by the need for a large amount of metabolic data. A first result presented in this paper is the identification of three types of cross-talks between PNs and FBA methods and their dependencies on available data. We exemplify our insights with the analysis of a pancreatic cancer model. We discuss how our reasoning framework provides a biologically and mathematically grounded decision making setting for the integration of regulatory, signalling, and metabolic networks and greatly increases model interpretability and reusability. We discuss how the parameters of PN and FBA models can be tuned and combined together so to highlight the computational effort needed to perform this task. We conclude with speculations and suggestions on this new promising research direction.

It is well known that the response of individuals to disease varies, either because of unpredictable exogenous events, such as possibly unknown environmental effects, or just because of endogenous factors, i.e. different genetic background. In particular, when a treatment effectiveness has to be validated, the individual variability should be taken into account by exploiting stochastic models. Relapsing Remitting Multiple Sclerosis (RRMS) is an unpredictable and complex disease, whose random behaviour perfectly fits the study with stochastic models. RRMS is the most common form of Multiple Sclerosis (MS), an immune-mediated inflammatory disease of the central nervous system, characterized by alternate episodes of symptom exacerbation (relapses) with periods of disease stability (remission). Several treatments were proposed to contrast the disease progression. Among these, Daclizumab initially exhibited promising results. However, due to the risk of serious side effects the treatment has been retired. We propose a stochastic and an hybrid extension, based on a generalization of the high level Petri Net formalism, of an existing model of Daclizumab effects on RRMS. The model is developed to investigate the complex mechanisms and unpredictable behaviour characterizing the RRMS disease and its relapsing, especially under the Daclizumab administration.

High-level Petri nets (HLPNs) are an expressive formalism well supported by a number of tools that automate the editing and the interactive simulation of models and some kinds of analytical techniques, mainly based on state-space exploration. Structural analysis of HLPNs is, however, a challenging task not yet adequately supported and it is often accomplished via the unfolding of an HLPN into a corresponding low-level Petri Net. An approach to derive a system of Ordinary Differential Equations (ODEs) from a Stochastic Symmetric Net (SSN) has been proposed a few years ago, based on the net's unfolding and subsequent grouping of similar equations. This method has been recently improved by providing an algorithm that directly derives a compact ODE system (from a partially unfolded net) in a symbolic way, through algebraic manipulation of SSN annotations. In this paper, we present the automation of the calculus of Symbolic ODEs (SODEs) for SSN models as a new module of SNexpression, a tool for the symbolic structural analysis of Symmetric Nets. An application of the tool/technique to a variant of a SIRS epidemic model including antibiotic resistance is also described.

BACKGROUND: Multiple Sclerosis (MS) is an immune-mediated inflammatory disease of the Central Nervous System (CNS) which damages the myelin sheath enveloping nerve cells thus causing severe physical disability in patients. Relapsing Remitting Multiple Sclerosis (RRMS) is one of the most common form of MS in adults and is characterized by a series of neurologic symptoms, followed by periods of remission. Recently, many treatments were proposed and studied to contrast the RRMS progression. Among these drugs, daclizumab (commercial name Zinbryta), an antibody tailored against the Interleukin-2 receptor of T cells, exhibited promising results, but its efficacy was accompanied by an increased frequency of serious adverse events. Manifested side effects consisted of infections, encephalitis, and liver damages. Therefore daclizumab has been withdrawn from the market worldwide. Another interesting case of RRMS regards its progression in pregnant women where a smaller incidence of relapses until the delivery has been observed. RESULTS: In this paper we propose a new methodology for studying RRMS, which we implemented in GreatSPN, a state-of-the-art open-source suite for modelling and analyzing complex systems through the Petri Net (PN) formalism. This methodology exploits: (a) an extended Colored PN formalism to provide a compact graphical description of the system and to automatically derive a set of ODEs encoding the system dynamics and (b) the Latin Hypercube Sampling with PRCC index to calibrate ODE parameters for reproducing the real behaviours in healthy and MS subjects.To show the effectiveness of such methodology a model of RRMS has been constructed and studied. Two different scenarios of RRMS were thus considered. In the former scenario the effect of the daclizumab administration is investigated, while in the latter one RRMS was studied in pregnant women. CONCLUSIONS: We propose a new computational methodology to study RRMS disease. Moreover, we show that model generated and calibrated according to this methodology is able to reproduce the expected behaviours.

Summary: Short reads sequencing technology has been used for more than a decade now. However, the analysis of RNAseq and ChIPseq data is still computational demanding and the simple access to raw data does not guarantee results reproducibility between laboratories. To address these two aspects, we developed SeqBox, a cheap, efficient and reproducible RNAseq/ChIPseq hardware/software solution based on NUC6I7KYK mini-PC (an Intel consumer game computer with a fast processor and a high performance SSD disk), and Docker container platform. In SeqBox the analysis of RNAseq and ChIPseq data is supported by a friendly GUI. This allows access to fast and reproducible analysis also to scientists with/without scripting experience. Availability and implementation: Docker container images, docker4seq package and the GUI are available at http://www.bioinformatica.unito.it/reproducibile.bioinformatics.html. Contact: [email protected]. Supplementary information: Supplementary data are available at Bioinformatics online.

BACKGROUND: Reproducibility of a research is a key element in the modern science and it is mandatory for any industrial application. It represents the ability of replicating an experiment independently by the location and the operator. Therefore, a study can be considered reproducible only if all used data are available and the exploited computational analysis workflow is clearly described. However, today for reproducing a complex bioinformatics analysis, the raw data and the list of tools used in the workflow could be not enough to guarantee the reproducibility of the results obtained. Indeed, different releases of the same tools and/or of the system libraries (exploited by such tools) might lead to sneaky reproducibility issues. RESULTS: To address this challenge, we established the Reproducible Bioinformatics Project (RBP), which is a non-profit and open-source project, whose aim is to provide a schema and an infrastructure, based on docker images and R package, to provide reproducible results in Bioinformatics. One or more Docker images are then defined for a workflow (typically one for each task), while the workflow implementation is handled via R-functions embedded in a package available at github repository. Thus, a bioinformatician participating to the project has firstly to integrate her/his workflow modules into Docker image(s) exploiting an Ubuntu docker image developed ad hoc by RPB to make easier this task. Secondly, the workflow implementation must be realized in R according to an R-skeleton function made available by RPB to guarantee homogeneity and reusability among different RPB functions. Moreover she/he has to provide the R vignette explaining the package functionality together with an example dataset which can be used to improve the user confidence in the workflow utilization. CONCLUSIONS: Reproducible Bioinformatics Project provides a general schema and an infrastructure to distribute robust and reproducible workflows. Thus, it guarantees to final users the ability to repeat consistently any analysis independently by the used UNIX-like architecture.

The aim of this paper is (i)to study breast cancer growth by mean of a mathematical model describing cell population dynamics during cancer growth, and (ii)to use this model to reproduce and explain experimental data. We started from a linear model describing cancer subpopulations evolution based on the Cancer Stem Cell (CSC) theory, and we added feedback mechanisms from the cell populations to mimic micro-environment effects in cancer growth. In details, we hypothesized two feedback mechanisms and we studied their effects both separately and combined together. In this way we obtained three new models that we tuned using data derived by TUBO Cancer cell line and describing the evolution of the total cell population and the subpopulations over time. Finally, we exploited these three models to understand which combination of feedback mechanisms better describe the experimental data.

BACKGROUND: Mantle Cell Lymphoma (MCL) is a B cell aggressive neoplasia accounting for about the 6% of all lymphomas. The most common molecular marker of clonality in MCL, as in other B lymphoproliferative disorders, is the ImmunoGlobulin Heavy chain (IGH) rearrangement, occurring in B-lymphocytes. The patient-specific IGH rearrangement is extensively used to monitor the Minimal Residual Disease (MRD) after treatment through the standardized Allele-Specific Oligonucleotides Quantitative Polymerase Chain Reaction based technique. Recently, several studies have suggested that the IGH monitoring through deep sequencing techniques can produce not only comparable results to Polymerase Chain Reaction-based methods, but also might overcome the classical technique in terms of feasibility and sensitivity. However, no standard bioinformatics tool is available at the moment for data analysis in this context. RESULTS: In this paper we present HashClone, an easy-to-use and reliable bioinformatics tool that provides B-cells clonality assessment and MRD monitoring over time analyzing data from Next-Generation Sequencing (NGS) technique. The HashClone strategy-based is composed of three steps: the first and second steps implement an alignment-free prediction method that identifies a set of putative clones belonging to the repertoire of the patient under study. In the third step the IGH variable region, diversity region, and joining region identification is obtained by the alignment of rearrangements with respect to the international ImMunoGenetics information system database. Moreover, a provided graphical user interface for HashClone execution and clonality visualization over time facilitate the tool use and the results interpretation. The HashClone performance was tested on the NGS data derived from MCL patients to assess the major B-cell clone in the diagnostic samples and to monitor the MRD in the real and artificial follow up samples. CONCLUSIONS: Our experiments show that in all the experimental settings, HashClone was able to correctly detect the major B-cell clones and to precisely follow them in several samples showing better accuracy than the state-of-art tool.

BACKGROUND: RNA-Seq provides remarkable power in the area of biomarkers discovery and disease characterization. Two crucial steps that affect RNA-Seq experiment results are Library Sample Preparation (LSP) and Bioinformatics Analysis (BA). This work describes an evaluation of the combined effect of LSP methods and BA tools in the detection of splice variants. RESULTS: Different LSPs (TruSeq unstranded/stranded, ScriptSeq, NuGEN) allowed the detection of a large common set of splice variants. However, each LSP also detected a small set of unique transcripts that are characterized by a low coverage and/or FPKM. This effect was particularly evident using the low input RNA NuGEN v2 protocol. A benchmark dataset, in which synthetic reads as well as reads generated from standard (Illumina TruSeq 100) and low input (NuGEN) LSPs were spiked-in was used to evaluate the effect of LSP on the statistical detection of alternative splicing events (AltDE). Statistical detection of AltDE was done using as prototypes for splice variant-quantification Cuffdiff2 and RSEM-EBSeq. As prototype for exon-level analysis DEXSeq was used. Exon-level analysis performed slightly better than splice variant-quantification approaches, although at most only 50% of the spiked-in transcripts was detected. The performances of both splice variant-quantification and exon-level analysis improved when raising the number of input reads. CONCLUSION: Data, derived from NuGEN v2, were not the ideal input for AltDE, especially when the exon-level approach was used. We observed that both splice variant-quantification and exon-level analysis performances were strongly dependent on the number of input reads. Moreover, the ribosomal RNA depletion protocol was less sensitive in detecting splicing variants, possibly due to the significant percentage of the reads mapping to non-coding transcripts.

Stochastic Symmetric Nets (SSNs) are a High-Level Stochastic Petri Net formalism which provides a parametric system description and an efficient analysis technique that exploit system symmetries to automatically aggregate its states. Even if significant reductions can be achieved in highly symmetric models, the reduced state space can still be too large to derive and/or solve the underlying stochastic process, so that Monte Carlo simulation and fluid approximation remain the only viable ways that need to be explored. In this paper, we contribute to this line of research by proposing a new approach based on fluid approximation to automatically derive from an SSN model a set of ordinary differential equations (ODEs) which mimic the system behavior, and by showing how the SSN formalism allows us to define an efficient translation method which reduces the size of the corresponding ODE system with an automatic exploitation of system symmetries. Additionally, some case studies are presented to show the effectiveness of the method and the relevance of its application in practical cases.

Abstract: This position paper introduces the concept of “artificial co-pilot ” (that is, a driver model), with a focus on driver’s oriented cognitive cars, in order to illustrate a new approach for future intelligent vehicles, which overcomes the limitations of nowadays models. The core consists in adopting the human cognitive frame-work for vehicles, following an artificial intelligent approach to take decisions. This paper illustrates in de-tails these concepts, as they are under development in the EU co-funded project HOLIDES. 1.

Abstract Summary: Chimera is a Bioconductor package that organizes, annotates, analyses and validates fusions reported by different fusion detection tools; current implementation can deal with output from bellerophontes, chimeraScan, deFuse, fusionCatcher, FusionFinder, FusionHunter, FusionMap, mapSplice, Rsubread, tophat-fusion and STAR. The core of Chimera is a fusion data structure that can store fusion events detected with any of the aforementioned tools. Fusions are then easily manipulated with standard R functions or through the set of functionalities specifically developed in Chimera with the aim of supporting the user in managing fusions and discriminating false-positive results. Availability and implementation: Chimera is implemented as a Bioconductor package in R. The package and the vignette can be downloaded at bioconductor.org. Contact: [email protected] Supplementary information: Supplementary data are available at Bioinformatics online.

In this paper, the Non-deterministic Repairable Fault Tree (NdRFT) formalism is proposed: it allows the modeling of failures of complex systems in addition to their repair processes. Its originality with respect to other Fault Tree extensions allows us to address repair strategy optimization problems: in an NdRFT model, the decision as to whether to start or not a given repair action is non-deterministic, so that all the possibilities are left open. The formalism is rather powerful, it allows: the specification of self-revealing events, the representation of components degradation, the choice among local repair, global repair, preventive maintenance, and the specification of the resources needed to start a repair action. The optimal repair strategy with respect to some relevant system state function, e.g. system unavailability, can then be computed by solving an optimization problem on a Markov Decision Process derived from the NdRFT. Such derivation is obtained by converting the NdRFT model into an intermediate formalism called Markov Decision Petri Net (MDPN). In the paper, the NdRFT syntax and semantics are formally described, together with the conversion rules to derive from the NdRFT the corresponding MDPN model. The application of NdRFT is illustrated through examples.

The Hybrid Automata Stochastic Logic (HASL) has been recently defined as a flexible way to express classical performance measures as well as more complex, path-based ones (generically called "HASL formulas"). The considered paths are executions of Generalized Stochastic Petri Nets (GSPN), which are an extension of the basic Petri net formalism to define discrete event stochastic processes. The computation of the HASL formulas for a GSPN model is demanded to the COSMOS tool, that applies simulation techniques to the formula computation. Stochastic Symmetric Nets (SSN) are a high level Petri net formalism, of the colored type, in which tokens can have an identity, and it is well known that colored Petri nets allow one to describe systems in a more compact and parametric form than basic (uncolored) Petri nets. In this paper we propose to extend HASL and COSMOS to support colors, so that performance formulas for SSN can be easily defined and evaluated. This requires a new definition of the logic, to ensure that colors are taken into account in a correct and useful manner, and a significant extension of the COSMOS tool.

BACKGROUND: RNA-seq has the potential to discover genes created by chromosomal rearrangements. Fusion genes, also known as "chimeras", are formed by the breakage and re-joining of two different chromosomes. It is known that chimeras have been implicated in the development of cancer. Few publications in the past showed the presence of fusion events also in normal tissue, but with very limited overlaps between their results. More recently, two fusion genes in normal tissues were detected using both RNA-seq and protein data.Due to heterogeneous results in identifying chimeras in normal tissue, we decided to evaluate the efficacy of state of the art fusion finders in detecting chimeras in RNA-seq data from normal tissues. RESULTS: We compared the performance of six fusion-finder tools: FusionHunter, FusionMap, FusionFinder, MapSplice, deFuse and TopHat-fusion. To evaluate the sensitivity we used a synthetic dataset of fusion-products, called positive dataset; in these experiments FusionMap, FusionFinder, MapSplice, and TopHat-fusion are able to detect more than 78% of fusion genes. All tools were error prone with high variability among the tools, identifying some fusion genes not present in the synthetic dataset. To better investigate the false discovery chimera detection rate, synthetic datasets free of fusion-products, called negative datasets, were used. The negative datasets have different read lengths and quality scores, which allow detecting dependency of the tools on both these features. FusionMap, FusionFinder, mapSplice, deFuse and TopHat-fusion were error-prone. Only FusionHunter results were free of false positive. FusionMap gave the best compromise in terms of specificity in the negative dataset and of sensitivity in the positive dataset. CONCLUSIONS: We have observed a dependency of the tools on read length, quality score and on the number of reads supporting each chimera. Thus, it is important to carefully select the software on the basis of the structure of the RNA-seq data under analysis. Furthermore, the sensitivity of chimera detection tools does not seem to be sufficient to provide results consistent with those obtained in normal tissues on the basis of fusion events extracted from published data.

BACKGROUND: Cancer stem cell theory suggests that cancers are derived by a population of cells named Cancer Stem Cells (CSCs) that are involved in the growth and in the progression of tumors, and lead to a hierarchical structure characterized by differentiated cell population. This cell heterogeneity affects the choice of cancer therapies, since many current cancer treatments have limited or no impact at all on CSC population, while they reveal a positive effect on the differentiated cell populations. RESULTS: In this paper we investigated the effect of vaccination on a cancer hierarchical structure through a multi-level model representing both population and molecular aspects. The population level is modeled by a system of Ordinary Differential Equations (ODEs) describing the cancer population's dynamics. The molecular level is modeled using the Petri Net (PN) formalism to detail part of the proliferation pathway. Moreover, we propose a new methodology which exploits the temporal behavior derived from the molecular level to parameterize the ODE system modeling populations. Using this multi-level model we studied the ErbB2-driven vaccination effect in breast cancer. CONCLUSIONS: We propose a multi-level model that describes the inter-dependencies between population and genetic levels, and that can be efficiently used to estimate the efficacy of drug and vaccine therapies in cancer models, given the availability of molecular data on the cancer driving force.

For the vast majority of species - including many economically or ecologically important organisms, progress in biological research is hampered due to the lack of a reference genome sequence. Despite recent advances in sequencing technologies, several factors still limit the availability of such a critical resource. At the same time, many research groups and international consortia have already produced BAC libraries and physical maps and now are in a position to proceed with the development of whole-genome sequences organized around a physical map anchored to a genetic map. We propose a BAC-by-BAC sequencing protocol that combines combinatorial pooling design and second-generation sequencing technology to efficiently approach denovo selective genome sequencing. We show that combinatorial pooling is a cost-effective and practical alternative to exhaustive DNA barcoding when preparing sequencing libraries for hundreds or thousands of DNA samples, such as in this case gene-bearing minimum-tiling-path BAC clones. The novelty of the protocol hinges on the computational ability to efficiently compare hundred millions of short reads and assign them to the correct BAC clones (deconvolution) so that the assembly can be carried out clone-by-clone. Experimental results on simulated data for the rice genome show that the deconvolution is very accurate, and the resulting BAC assemblies have high quality. Results on real data for a gene-rich subset of the barley genome confirm that the deconvolution is accurate and the BAC assemblies have good quality. While our method cannot provide the level of completeness that one would achieve with a comprehensive whole-genome sequencing project, we show that it is quite successful in reconstructing the gene sequences within BACs. In the case of plants such as barley, this level of sequence knowledge is sufficient to support critical end-point objectives such as map-based cloning and marker-assisted breeding.

In this paper we propose an improved BDD approach to the network reliability analysis, that allows the user to compute an exact solution or an approximation based on reliability bounds when network complexity makes the former solution practically impossible. To this purpose, a new algorithm for encoding the connectivity graph on a Binary Decision Diagram (BDD) has been developed; it reduces the computation memory peak with respect to previous approaches based on the same type of data structure without increasing the execution time, and allows us also to derive from a subset of the minpaths/mincuts a lower/upper bound of the network reliability, so that the quality of the obtained approximation can be estimated. Finally, a fair and detailed comparison between our approach and another state of the art approach presented in the literature is documented through a set of benchmarks.

In this paper we propose a methodology for the modeling and analysis of ad hoc networks composed by a large number of nodes moving among geographical regions. This methodology uses compositional construction of stochastic Petri nets (SPN) for building the model which allows for specifying the model and the required performance indices at a high level of abstraction. As our aim is to consider real scenarios with several geographical regions and non-trivial user behavior in each region, the size of the state space of the model can easily grow too large to analyze with exact analytical approaches or even with simulation. For this reason, we propose to carry out the analysis by constructing the mean field approximation of the behavior of the SPN. The approximation is provided by a set of ordinary differential equations (ODE) that can be derived automatically from the SPN and can be solved numerically with low computational effort even for large models. The methodology is illustrated on a case study, modeling application spreading in a mobile environment. It will be shown that the approximate results obtained by the mean field approach capture well the behavior of the system.

The increasing demand for customer centric evaluation of systems, mostly related with the assessment of the quality of service that they can deliver, requires the development of techniques properly designed to model and to study the movement of specific entities generically referred to as "customers". Stochastic Well-Formed Net(SWN) are naturally suited for the representation of systems in which "customers" of different categories compete for the use of common resources. Color classes of SWN are easily associated with these different categories, leaving to the peculiar features of the formalism the possibility of exploiting all the symmetries existing into the representation for the efficient and effective computation of the measures of interest. Within this application context, the computation of first passage time distribution measures in Stochastic Well-Formed Net (SWN) is becoming of primary interest. Customers however are not primitive entities in the formalism and an approach similar to that previously developed for Generalized Stochastic Petri Nets (GSPN) is suggested to overcome this problem in which P-semiflows are used to identify the circulating "customers". In this paper we propose an original algorithm for computing some P-semiflows of colored PNs (in particular of SWNs) in parametric form by exploiting the peculiarities of the objective of this investigation, and extend the customer centric first passage time computation approach previously developed for GSPNs, to make it suitable for SWN models. Moreover, the paper proposes an enhancement of the SWN notation in order to provide a way to ease the modeler in the specification of customer scheduling policies that may affect the computation of first passage time distributions. This extension, inspired by Queueing Petri Nets, adds to SWN some "syntactic sugar" that allows to include in the model queueing places which are automatically replaced by appropriate submodels, before solving the model.

This paper presents an extension of the generalized stochastic Petri net (GSPN) formalism that enables the computation of first passage time distributions. The tagged customer technique typical of queuing networks is adapted to the GSPN context by providing a formal definition and an automatic computation of the groups of tokens that can be identified as customers, i.e. classes of homogeneous entities behaving in a similar manner. Passage times are identified through the concept of events that correspond to the firing of transitions placed at the boundaries of a subnet. The extended model obtained with this specifications is translated into an ordinary GSPN by isolating a customer from the group and highlighting its path through the net thus obtaining a representation suited for the passage time analysis. Proofs are provided to show the equivalence between these models with respect to their steady-state distributions. An important and original aspect treated in this paper is the possibility of specifying several scheduling policies of tokens at places, an information not present in ordinary GSPN models, but that is vital for the precise computation of first passage time distributions as shown by a few results computed for a simple Flexible Manufacturing application.

Non deterministic Repairable Fault Trees (NdRFT) are a recently proposed modeling formalism for the study of optimal repair strategies: they are based on the widely adopted Fault Tree formalism, but in addition to the failure modes, NdRFTs allow to define possible repair actions. In a previous pa per the formalism has been introduced together with an analysis method and a tool allowing to automatically derive the best repair strategy to be applied in each state. The analysis technique is based on the generation and solution of a Markov Decision Process. In this paper we present an extension, ParNdRFT, that allows to exploit the presence of redundancy to reduce the complexity of the model and of the analysis. It is based on the translation of the ParNdRFT in to a Markov Decision Well-Formed Net, i.e. a model specified by means of an High Level Petri Net formalism. The translated model can be efficiently solved thanks to existing algorithms that generate a reduced state space automatically exploiting the model symmetries.