Suggestions and Comparisons of Two Algorithms for the Simplification of Bluetooth Sensor Data in Traffic Cordons.

Bluetooth sensors in intelligent transportation systems possess extensive coverage and access to a large number of identity (ID) data, but they cannot distinguish between vehicles and persons. This study aims to classify and differentiate raw data collected from Bluetooth sensors positioned between various origin-destination (i-j) points into vehicles and persons and to determine their distribution ratios. To reduce data noise, two different filtering algorithms are proposed. The first algorithm employs time series simplification based on Simple Moving Average (SMA) and threshold models, which are tools of statistical analysis. The second algorithm is rule-based, using speed data of Bluetooth devices derived from sensor data to provide a simplification algorithm. The study area was the Historic Peninsula Traffic Cord Region of Istanbul, utilizing data from 39 sensors in the region. As a result of time-based filtering, the ratio of person ID addresses for Bluetooth devices participating in circulation in the region was found to be 65.57% (397,799 person IDs), while the ratio of vehicle ID addresses was 34.43% (208,941 vehicle IDs). In contrast, the rule-based algorithm based on speed data found that the ratio of vehicle ID addresses was 35.82% (389,392 vehicle IDs), while the ratio of person ID addresses was 64.17% (217,348 person IDs). The Jaccard similarity coefficient was utilized to identify similarities in the data obtained from the applied filtering approaches, yielding a coefficient (J) of 0.628. The identity addresses of the vehicles common throughout the two date sets which are obtained represent the sampling size for traffic measurements.

Micro level analyses of environmentally disastrous urbanization in Bangalore.

Indian metropolitan (tier I) cities have been undergoing rapid urbanization during the post-globalization era with the unprecedented market interventions, which have led to the rapid land cover changes affecting the ecology, climate, hydrology, and local environment. The unplanned urbanization has given way to the dispersed, haphazard growth at the city outskirts with the lack of basic amenities and infrastructure as the planners lack advance information of sprawl regions. This has necessitated understanding and visualization of urbanization patterns for planning towards sustainable cities. The analyses of urban dynamics during 1973-2017 using temporal remote sensing data reveal 1028% increase in urban area with the decline of 88% vegetation and 79% of water bodies. Consequences of the unplanned urbanization are the increase in greenhouse gas emissions, decline in vegetation cover, loss of groundwater table (from 28 to 300 m), contamination of water sources, increase in land surface temperature, increase in disease vectors, etc. An attempt is made to understand the implications of unplanned growth at the micro level by considering the prime growth poles such as Peenya Industrial Estate (PIE), Whitefield (WF), Bangalore South Region (BSR). The spatial analyses reveal the decline of vegetation and open spaces with intense urbanization of 86.35% (in BSR), 87.39% (PIE) and 81.61% (WF) in 2017. WF witnessed the drastic transformation from agrarian ecosystem to a concrete jungle during the past four decades. Spatial patterns of urbanization were assessed through the landscape metrics and rule-based modeling which confirms intense urbanization with single class dominance. Specifically, NP metrics depicts PIE region had sprawl growth till 2003 with numerous patches and is transformed by 2017 it has become to a single dense urban patch. This necessitates appropriate planning strategies to mitigate further erosion of environmental resources and ensure clean air, water, and environment to all residents.

Generalizing Gillespie's Direct Method to Enable Network-Free Simulations.

Gillespie's direct method for stochastic simulation of chemical kinetics is a staple of computational systems biology research. However, the algorithm requires explicit enumeration of all reactions and all chemical species that may arise in the system. In many cases, this is not feasible due to the combinatorial explosion of reactions and species in biological networks. Rule-based modeling frameworks provide a way to exactly represent networks containing such combinatorial complexity, and generalizations of Gillespie's direct method have been developed as simulation engines for rule-based modeling languages. Here, we provide both a high-level description of the algorithms underlying the simulation engines, termed network-free simulation algorithms, and how they have been applied in systems biology research. We also define a generic rule-based modeling framework and describe a number of technical details required for adapting Gillespie's direct method for network-free simulation. Finally, we briefly discuss potential avenues for advancing network-free simulation and the role they continue to play in modeling dynamical systems in biology.

Small drinking water systems under spatiotemporal water quality variability: a risk-based performance benchmarking framework.

Traditional approaches for benchmarking drinking water systems are binary, based solely on the compliance and/or non-compliance of one or more water quality performance indicators against defined regulatory guidelines/standards. The consequence of water quality failure is dependent on location within a water supply system as well as time of the year (i.e., season) with varying levels of water consumption. Conventional approaches used for water quality comparison purposes fail to incorporate spatiotemporal variability and degrees of compliance and/or non-compliance. This can lead to misleading or inaccurate performance assessment data used in the performance benchmarking process. In this research, a hierarchical risk-based water quality performance benchmarking framework is proposed to evaluate small drinking water systems (SDWSs) through cross-comparison amongst similar systems. The proposed framework (R WQI framework) is designed to quantify consequence associated with seasonal and location-specific water quality issues in a given drinking water supply system to facilitate more efficient decision-making for SDWSs striving for continuous performance improvement. Fuzzy rule-based modelling is used to address imprecision associated with measuring performance based on singular water quality guidelines/standards and the uncertainties present in SDWS operations and monitoring. This proposed R WQI framework has been demonstrated using data collected from 16 SDWSs in Newfoundland and Labrador and Quebec, Canada, and compared to the Canadian Council of Ministers of the Environment WQI, a traditional, guidelines/standard-based approach. The study found that the R WQI framework provides an in-depth state of water quality and benchmarks SDWSs more rationally based on the frequency of occurrence and consequence of failure events.

Predicting gene-level sensitivity to JAK-STAT signaling perturbation using a mechanistic-to-machine learning framework.

The Janus kinase (JAK)-signal transducer and activator of transcription (STAT) pathway integrates complex cytokine signals via a limited number of molecular components, inspiring numerous efforts to clarify the diversity and specificity of STAT transcription factor function. We developed a computational framework to make global cytokine-induced gene predictions from STAT phosphorylation dynamics, modeling macrophage responses to interleukin (IL)-6 and IL-10, which signal through common STATs, but with distinct temporal dynamics and contrasting functions. Our mechanistic-to-machine learning model identified cytokine-specific genes associated with late pSTAT3 time frames and a preferential pSTAT1 reduction upon JAK2 inhibition. We predicted and validated the impact of JAK2 inhibition on gene expression, identifying genes that were sensitive or insensitive to JAK2 variation. Thus, we successfully linked STAT signaling dynamics to gene expression to support future efforts targeting pathology-associated STAT-driven gene sets. This serves as a first step in developing multi-level prediction models to understand and perturb gene expression outputs from signaling systems. A record of this paper's transparent peer review process is included in the supplemental information.

The solubility product extends the buffering concept to heterotypic biomolecular condensates.

Biomolecular condensates are formed by liquid-liquid phase separation (LLPS) of multivalent molecules. LLPS from a single ("homotypic") constituent is governed by buffering: above a threshold, free monomer concentration is clamped, with all added molecules entering the condensed phase. However, both experiment and theory demonstrate that buffering fails for the concentration dependence of multicomponent ("heterotypic") LLPS. Using network-free stochastic modeling, we demonstrate that LLPS can be described by the solubility product constant (Ksp): the product of free monomer concentrations, accounting for the ideal stoichiometries governed by the valencies, displays a threshold above which additional monomers are funneled into large clusters; this reduces to simple buffering for homotypic systems. The Ksp regulates the composition of the dilute phase for a wide range of valencies and stoichiometries. The role of Ksp is further supported by coarse-grained spatial particle simulations. Thus, the solubility product offers a general formulation for the concentration dependence of LLPS.

Receptor/Raft Ratio Is a Determinant for LRP6 Phosphorylation and WNT/ß-Catenin Signaling.

Microdomains or lipid rafts greatly affect the distribution of proteins and peptides in the membrane and play a vital role in the formation and activation of receptor/protein complexes. A prominent example for the decisive impact of lipid rafts on signaling is LRP6, whose localization to the same lipid rafts domain as the kinase CK1γ is crucial for its successful phosphorylation and the subsequent activation of the signalosome, hence WNT/ß-catenin signaling. However, according to various experimental measurements, approximately 25 to 35 % of the cell plasma membrane is covered by nanoscopic raft domains with diameters ranging between 10 to 200 nm. Extrapolating/Translating these values to the membrane of a "normal sized" cell yields a raft abundance, that, by far, outnumbers the membrane-associated pathway components of most individual signaling pathway, such as receptor and kinases. To analyze whether and how the quantitative ratio between receptor and rafts affects LRP6 phosphorylation and WNT/ß-catenin pathway activation, we present a computational modeling study, that for the first time employs realistic raft numbers in a compartment-based pathway model. Our simulation experiments indicate, that for receptor/raft ratios smaller than 1, i.e., when the number of raft compartments clearly exceeds the number of pathway specific membrane proteins, we observe significant decrease in LRP6 phosphorylation and downstream pathway activity. Our results suggest that pathway specific targeting and sorting mechanism are required to significantly narrow down the receptor/raft ratio and to enable the formation of the LRP6 signalosome, hence signaling.

Systems biology markup language (SBML) level 3 package: multistate, multicomponent and multicompartment species, version 1, release 2.

Rule-based modeling is an approach that permits constructing reaction networks based on the specification of rules for molecular interactions and transformations. These rules can encompass details such as the interacting sub-molecular domains and the states and binding status of the involved components. Conceptually, fine-grained spatial information such as locations can also be provided. Through "wildcards" representing component states, entire families of molecule complexes sharing certain properties can be specified as patterns. This can significantly simplify the definition of models involving species with multiple components, multiple states, and multiple compartments. The systems biology markup language (SBML) Level 3 Multi Package Version 1 extends the SBML Level 3 Version 1 core with the "type" concept in the Species and Compartment classes. Therefore, reaction rules may contain species that can be patterns and exist in multiple locations. Multiple software tools such as Simmune and BioNetGen support this standard that thus also becomes a medium for exchanging rule-based models. This document provides the specification for Release 2 of Version 1 of the SBML Level 3 Multi package. No design changes have been made to the description of models between Release 1 and Release 2; changes are restricted to the correction of errata and the addition of clarifications.

Using Mechanistic Models for Analysis of Proteomic Data.

A goal of systems biology is to develop an integrated picture of how the myriad components of a biological system work together to produce responses to environmental inputs. Achieving this goal requires (1) assembling a list of the component parts of a cellular regulatory system, and (2) understanding how the connections between these components enable information processing. To work toward these ends, a number of methods have matured in parallel. The compilation of a cellular parts list has been accelerated by the advent of omics technologies, which enable simultaneous characterization of a large collection of biomolecules. A particular type of omics technology that is useful for understanding protein-protein interaction networks is proteomics, which can give information about a number of dimensions of the state of the cell's proteins: quantification of protein abundances within the cell, characterization of the posttranslational modification state of the proteome through phosphopeptide enrichment, and identification of protein-protein interactions through co-immunoprecipitation. Mathematical models can be useful in analyzing proteomic data.

RKappa: Software for Analyzing Rule-Based Models.

RKappa is a framework for the development, simulation, and analysis of rule-based models within the mature statistically empowered R environment. It is designed for model editing, parameter identification, simulation, sensitivity analysis, and visualization. The framework is optimized for high-performance computing platforms and facilitates analysis of large-scale systems biology models where knowledge of exact mechanisms is limited and parameter values are uncertain.The RKappa software is an open-source (GLP3 license) package for R, which is freely available online ( https://github.com/lptolik/R4Kappa ).

Using rxncon to Develop Rule-Based Models.

We present a protocol for building, validating, and simulating models of signal transduction networks. These networks are challenging modeling targets due to the combinatorial complexity and sparse data, which have made it a major challenge even to formalize the current knowledge. To address this, the community has developed methods to model biomolecular reaction networks based on site dynamics. The strength of this approach is that reactions and states can be defined at variable resolution, which makes it possible to adapt the model resolution to the empirical data. This improves both scalability and accuracy, making it possible to formalize large models of signal transduction networks. Here, we present a method to build and validate large models of signal transduction networks. The workflow is based on rxncon, the reaction-contingency language. In a five-step process, we create a mechanistic network model, convert it into an executable Boolean model, use the Boolean model to evaluate and improve the network, and finally export the rxncon model into a rule-based format. We provide an introduction to the rxncon language and an annotated, step-by-step protocol for the workflow. Finally, we create a small model of the insulin signaling pathway to illustrate the protocol, together with some of the challenges-and some of their solutions-in modeling signal transduction.

Mechanistic Models of Cellular Signaling, Cytokine Crosstalk, and Cell-Cell Communication in Immunology.

The cells of the immune system respond to a great variety of different signals that frequently reach them simultaneously. Computational models of signaling pathways and cellular behavior can help us explore the biochemical mechanisms at play during such responses, in particular when those models aim at incorporating molecular details of intracellular reaction networks. Such detailed models can encompass hypotheses about the interactions among molecular binding domains and how these interactions are modulated by, for instance, post-translational modifications, or steric constraints in multi-molecular complexes. In this way, the models become formal representations of mechanistic immunological hypotheses that can be tested through quantitative simulations. Due to the large number of parameters (molecular abundances, association-, dissociation-, and enzymatic transformation rates) the goal of simulating the models can, however, in many cases no longer be the fitting of particular parameter values. Rather, the simulations perform sweeps through parameter space to test whether a model can account for certain experimentally observed features when allowing the parameter values to vary within experimentally determined or physiologically reasonable ranges. We illustrate how this approach can be used to explore possible mechanisms of immunological pathway crosstalk. Probing the input-output behavior of mechanistic pathway models through systematic simulated variations of receptor stimuli will soon allow us to derive cell population behavior from single-cell models, thereby bridging a scale gap that currently still is frequently addressed through heuristic phenomenological multi-scale models.

Using RuleBuilder to Graphically Define and Visualize BioNetGen-Language Patterns and Reaction Rules.

RuleBuilder is a tool for drawing graphs that can be represented by the BioNetGen language (BNGL), which is used to formulate mathematical, rule-based models of biochemical systems. BNGL provides an intuitive plain text, or string, representation of such systems, which is based on a graphical formalism. Reactions are defined in terms of graph-rewriting rules that specify the necessary intrinsic properties of the reactants, a transformation, and a rate law. Rules also contain contextual constraints that restrict application of the rule. In some cases, the specification of contextual constraints can be verbose, making a rule difficult to read. RuleBuilder is designed to ease the task of reading and writing individual reaction rules or other BNGL patterns required for model formulation. The software assists in the reading of existing models by converting BNGL strings of interest into a graph-based representation composed of nodes and edges. RuleBuilder also enables the user to construct de novo a visual representation of BNGL strings using drawing tools available in its interface. As objects are added to the drawing canvas, the corresponding BNGL string is generated on the fly, and objects are similarly drawn on the fly as BNGL strings are entered into the application. RuleBuilder thus facilitates construction and interpretation of rule-based models.

Multi-Level Modeling and Simulation of Cellular Systems: An Introduction to ML-Rules.

ML-Rules is a rule-based language for multi-level modeling and simulation. ML-Rules supports dynamic nesting of entities and applying arbitrary functions on entity attributes and content, as well as for defining kinetics of reactions. This allows describing and simulating complex cellular dynamics operating at different organizational levels, e.g., to combine intra-, inter-, and cellular dynamics, like the proliferation of cells, or to include compartmental dynamics like merging and splitting of mitochondria or endocytosis. The expressiveness of the language is bought with additional efforts in executing ML-Rules models. Therefore, various simulators have been developed from which the user and automatic procedures can select. The experiment specification language SESSL facilitates design, execution, and reuse of simulation experiments. The chapter illuminates the specific features of ML-Rules as a rule-based modeling language, the implications for an efficient execution, and shows ML-Rules at work.

Rule-Based Modeling Using Wildcards in the Smoldyn Simulator.

Many biological molecules exist in multiple variants, such as proteins with different posttranslational modifications, DNAs with different sequences, and phospholipids with different chain lengths. Representing these variants as distinct species, as most biochemical simulators do, leads to the problem that the number of species, and chemical reactions that interconvert them, typically increase combinatorially with the number of ways that the molecules can vary. This can be alleviated by "rule-based modeling methods," in which software generates the chemical reaction network from relatively simple "rules." This chapter presents a new approach to rule-based modeling. It is based on wildcards that match to species names, much as wildcards can match to file names in computer operating systems. It is much simpler to use than the formal rule-based modeling approaches developed previously but can lead to unintended consequences if not used carefully. This chapter demonstrates rule-based modeling with wildcards through examples for signaling systems, protein complexation, polymerization, nucleic acid sequence copying and mutation, the "SMILES" chemical notation, and others. The method is implemented in Smoldyn, a spatial and stochastic biochemical simulator, for both generate-first and on-the-fly expansion, meaning whether the reaction network is generated before or during the simulation.

MCell-R: A Particle-Resolution Network-Free Spatial Modeling Framework.

Spatial heterogeneity can have dramatic effects on the biochemical networks that drive cell regulation and decision-making. For this reason, a number of methods have been developed to model spatial heterogeneity and incorporated into widely used modeling platforms. Unfortunately, the standard approaches for specifying and simulating chemical reaction networks become untenable when dealing with multistate, multicomponent systems that are characterized by combinatorial complexity. To address this issue, we developed MCell-R, a framework that extends the particle-based spatial Monte Carlo simulator, MCell, with the rule-based model specification and simulation capabilities provided by BioNetGen and NFsim. The BioNetGen syntax enables the specification of biomolecules as structured objects whose components can have different internal states that represent such features as covalent modification and conformation and which can bind components of other molecules to form molecular complexes. The network-free simulation algorithm used by NFsim enables efficient simulation of rule-based models even when the size of the network implied by the biochemical rules is too large to enumerate explicitly, which frequently occurs in detailed models of biochemical signaling. The result is a framework that can efficiently simulate systems characterized by combinatorial complexity at the level of spatially resolved individual molecules over biologically relevant time and length scales.

A Step-by-Step Guide to Using BioNetFit.

BioNetFit is a software tool designed for solving parameter identification problems that arise in the development of rule-based models. It solves these problems through curve fitting (i.e., nonlinear regression). BioNetFit is compatible with deterministic and stochastic simulators that accept BioNetGen language (BNGL)-formatted files as inputs, such as those available within the BioNetGen framework. BioNetFit can be used on a laptop or stand-alone multicore workstation as well as on many Linux clusters, such as those that use the Slurm Workload Manager to schedule jobs. BioNetFit implements a metaheuristic population-based global optimization procedure, an evolutionary algorithm (EA), to minimize a user-defined objective function, such as a residual sum of squares (RSS) function. BioNetFit also implements a bootstrapping procedure for determining confidence intervals for parameter estimates. Here, we provide step-by-step instructions for using BioNetFit to estimate the values of parameters of a BNGL-encoded model and to define bootstrap confidence intervals. The process entails the use of several plain-text files, which are processed by BioNetFit and BioNetGen. In general, these files include (1) one or more EXP files, which each contains (experimental) data to be used in parameter identification/bootstrapping; (2) a BNGL file containing a model section, which defines a (rule-based) model, and an actions section, which defines simulation protocols that generate GDAT and/or SCAN files with model predictions corresponding to the data in the EXP file(s); and (3) a CONF file that configures the fitting/bootstrapping job and that defines algorithmic parameter settings.

Rule-Based Models and Applications in Biology.

Complex systems are governed by dynamic processes whose underlying causal rules are difficult to unravel. However, chemical reactions, molecular interactions, and many other complex systems can be usually represented as concentrations or quantities that vary over time, which provides a framework to study these dynamic relationships. An increasing number of tools use these quantifications to simulate dynamically complex systems to better understand their underlying processes. The application of such methods covers several research areas from biology and chemistry to ecology and even social sciences.In the following chapter, we introduce the concept of rule-based simulations based on the Stochastic Simulation Algorithm (SSA) as well as other mathematical methods such as Ordinary Differential Equations (ODE) models to describe agent-based systems. Besides, we describe the mathematical framework behind Kappa (κ), a rule-based language for the modeling of complex systems, and some extensions for spaßtial models implemented in PISKaS (Parallel Implementation of a Spatial Kappa Simulator). To facilitate the understanding of these methods, we include examples of how these models can be used to describe population dynamics in a simple predator-prey ecosystem or to simulate circadian rhythm changes.

SBML Level 3 package: Multistate, Multicomponent and Multicompartment Species, Version 1, Release 1.

Rule-based modeling is an approach that permits constructing reaction networks based on the specification of rules for molecular interactions and transformations. These rules can encompass details such as the interacting sub-molecular domains (components) and the states such as phosphorylation and binding status of the involved components. Fine-grained spatial information such as the locations of the molecular components relative to a membrane (e.g. whether a modeled molecular domain is embedded into the inner leaflet of the cellular plasma membrane) can also be provided. Through wildcards representing component states entire families of molecule complexes sharing certain properties can be specified as patterns. This can significantly simplify the definition of models involving species with multiple components, multiple states and multiple compartments. The SBML Level 3 Multi Package (Multistate, Multicomponent and Multicompartment Species Package for SBML Level 3) extends the SBML Level 3 core with the "type" concept in the Species and Compartment classes and therefore reaction rules may contain species that can be patterns and be in multiple locations in reaction rules. Multiple software tools such as Simmune and BioNetGen support the SBML Level 3 Multi package that thus also becomes a medium for exchanging rule-based models.

An Overview of Network-Based and -Free Approaches for Stochastic Simulation of Biochemical Systems.

Stochastic simulation has been widely used to model the dynamics of biochemical reaction networks. Several algorithms have been proposed that are exact solutions of the chemical master equation, following the work of Gillespie. These stochastic simulation approaches can be broadly classified into two categories: network-based and -free simulation. The network-based approach requires that the full network of reactions be established at the start, while the network-free approach is based on reaction rules that encode classes of reactions, and by applying rule transformations, it generates reaction events as they are needed without ever having to derive the entire network. In this study, we compare the efficiency and limitations of several available implementations of these two approaches. The results allow for an informed selection of the implementation and methodology for specific biochemical modeling applications.