An incremental modular technique for checking LTL-X properties on Petri nets

Model-checking is a powerful and widespread technique for the verification of finite state concurrent systems. However, the main hindrance for wider application of this technique is the well-known state explosion problem. Modular verification is a promising natural approach to tackle this problem. It is based on the "divide and conquer" principle and aims at deducing the properties of the system from those of its components analysed in isolation. Unfortunately, several issues make the use of modular verification techniques difficult in practice. First, deciding how to partition the system into components is not trivial and can have a significant impact on the resources needed for verification. Second, when model-checking a component in isolation, how should the environment of this component be described? In this paper, we address these problems in the framework of model-checking LTL\X action-based properties on Petri nets. We propose an incremental and modular verification approach where the system model is partitioned according to the actions occurring in the property to be verified and where the environment of a component is taken into account using the linear place invariants of the system

Identification of a novel zinc metalloprotease through a global analysis of clostridium difficile extracellular proteins

Clostridium difficile is a major cause of infectious diarrhea worldwide. Although the cell surface proteins are recognized to be important in clostridial pathogenesis, biological functions of only a few are known. Also, apart from the toxins, proteins exported by C. difficile into the extracellular milieu have been poorly studied. In order to identify novel extracellular factors of C. difficile, we analyzed bacterial culture supernatants prepared from clinical isolates, 630 and R20291, using liquid chromatography-tandem mass spectrometry. The majority of the proteins identified were non-canonical extracellular proteins. These could be largely classified into proteins associated to the cell wall (including CWPs and extracellular hydrolases), transporters and flagellar proteins. Seven unknown hypothetical proteins were also identified. One of these proteins, CD630_28300, shared sequence similarity with the anthrax lethal factor, a known zinc metallopeptidase. We demonstrated that CD630_28300 (named Zmp1) binds zinc and is able to cleave fibronectin and fibrinogen in vitro in a zinc-dependent manner. Using site-directed mutagenesis, we identified residues important in zinc binding and enzymatic activity. Furthermore, we demonstrated that Zmp1 destabilizes the fibronectin network produced by human fibroblasts. Thus, by analyzing the exoproteome of C. difficile, we identified a novel extracellular metalloprotease that may be important in key steps of clostridial pathogenesis

Component-wise incremental LTL model checking

Efficient symbolic and explicit-state model checking approaches have been developed for the verification of linear time temporal logic (LTL) properties. Several attempts have been made to combine the advantages of the various algorithms. Model checking LTL properties usually poses two challenges: one must compute the synchronous product of the state space and the automaton model of the desired property, then look for counterexamples that is reduced to finding strongly connected components (SCCs) in the state space of the product. In case of concurrent systems, where the phenomenon of state space explosion often prevents the successful verification, the so-called saturation algorithm has proved its efficiency in state space exploration. This paper proposes a new approach that leverages the saturation algorithm both as an iteration strategy constructing the product directly, as well as in a new fixed-point computation algorithm to find strongly connected components on-the-fly by incrementally processing the components of the model. Complementing the search for SCCs, explicit techniques and component-wise abstractions are used to prove the absence of counterexamples. The resulting on-the-fly, incremental LTL model checking algorithm proved to scale well with the size of models, as the evaluation on models of the Model Checking Contest suggests

Increased sporulation underpins adaptation of Clostridium difficile strain 630 to a biologically–relevant faecal environment, with implications for pathogenicity

Abstract Clostridium difficile virulence is driven primarily by the processes of toxinogenesis and sporulation, however many in vitro experimental systems for studying C. difficile physiology have arguably limited relevance to the human colonic environment. We therefore created a more physiologically–relevant model of the colonic milieu to study gut pathogen biology, incorporating human faecal water (FW) into growth media and assessing the physiological effects of this on C. difficile strain 630. We identified a novel set of C. difficile–derived metabolites in culture supernatants, including hexanoyl– and pentanoyl–amino acid derivatives by LC-MSn. Growth of C. difficile strain 630 in FW media resulted in increased cell length without altering growth rate and RNA sequencing identified 889 transcripts as differentially expressed (p < 0.001). Significantly, up to 300–fold increases in the expression of sporulation–associated genes were observed in FW media–grown cells, along with reductions in motility and toxin genes’ expression. Moreover, the expression of classical stress–response genes did not change, showing that C. difficile is well–adapted to this faecal milieu. Using our novel approach we have shown that interaction with FW causes fundamental changes in C. difficile biology that will lead to increased disease transmissibility

An incremental modular technique for checking LTL-X properties on Petri nets

Model-checking is a powerful and widespread technique for the verification of finite state concurrent systems. However, the main hindrance for wider application of this technique is the well-known state explosion problem. Modular verification is a promising natural approach to tackle this problem. It is based on the "divide and conquer" principle and aims at deducing the properties of the system from those of its components analysed in isolation. Unfortunately, several issues make the use of modular verification techniques difficult in practice. First, deciding how to partition the system into components is not trivial and can have a significant impact on the resources needed for verification. Second, when model-checking a component in isolation, how should the environment of this component be described? In this paper, we address these problems in the framework of model-checking LTL\X action-based properties on Petri nets. We propose an incremental and modular verification approach where the system model is partitioned according to the actions occurring in the property to be verified and where the environment of a component is taken into account using the linear place invariants of the system

Behavioral technique for workflow abstraction and matching

This work is in line with the CoopFlow approach dedicated for workflow advertisement, interconnection, and cooperation in virtual organizations. In order to advertise workflows into a registry, we present in this paper a novel method to abstract behaviors of workflows into symbolic observation graphs (SOG). We present in addition an efficient algorithm for SOG matching, which is used for interconnecting workflows1

A Formal Model for Business Process Configuration Verification Supporting OR-Join Semantics

International audienceIn today's industries, similar process models are typically reused in different application contexts. These models result in a number of process model variants sharing several commonalities and exhibiting some variations. Configurable process models came to represent and group these variants in a generic manner. These processes are configured according to a specific context through configurable elements. Considering the large number of possible variants as well as the potentially complex configurable process, the configuration may be a tedious task and errors may lead to serious behavioral issues. Since achieving configuration in a correct manner has become of paramount importance, the analysts undoubtedly need assistance and guidance in configuring process variants. In this work, we propose a formal behavioral model based on the Symbolic Observation Graph (SOG) allowing to find the set of deadlock-free configuration choices while avoiding the well-known state-space explosion problem and considering loops and OR-join semantics. These choices are used to support business analysts in deriving deadlock-free variant

Multiple factors modulate biofilm formation by the anaerobic pathogen Clostridium difficile

Bacteria within biofilms are protected from multiple stresses, including immune responses and antimicrobial agents. The biofilm-forming ability of bacterial pathogens has been associated with increased antibiotic resistance and chronic recurrent infections. Although biofilms have been well studied for several gut pathogens, little is known about biofilm formation by anaerobic gut species. The obligate anaerobe Clostridium difficile causes C. difficile infection (CDI), a major health care-associated problem primarily due to the high incidence of recurring infections. C. difficile colonizes the gut when the normal intestinal microflora is disrupted by antimicrobial agents; however, the factors or processes involved in gut colonization during infection remain unclear. We demonstrate that clinical C. difficile strains, i.e., strain 630 and the hypervirulent strain R20291, form structured biofilms in vitro, with R20291 accumulating substantially more biofilm. Microscopic and biochemical analyses show multiple layers of bacteria encased in a biofilm matrix containing proteins, DNA, and polysaccharide. Employing isogenic mutants, we show that virulence-associated proteins, Cwp84, flagella, and a putative quorum-sensing regulator, LuxS, are all required for maximal biofilm formation by C. difficile. Interestingly, a mutant in Spo0A, a transcription factor that controls spore formation, was defective for biofilm formation, indicating a possible link between sporulation and biofilm formation. Furthermore, we demonstrate that bacteria in clostridial biofilms are more resistant to high concentrations of vancomycin, a drug commonly used for treatment of CDI. Our data suggest that biofilm formation by C. difficile is a complex multifactorial process and may be a crucial mechanism for clostridial persistence in the host