Julie Chapron

Xavier Boucher
email: boucher@emse.fr

Patrick Burlat

Pierre Lebrun

Stmicroelectronics Boucher

Analysis of Organisational Dependency for Urbanism of Information Systems

Keywords: CLUSTERING, ENTERPRISE MODELLING, INFORMATION SYSTEMS user): dependencies, coordination dependencies, processes, clustering, evolution management, coordination

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Introduction

In the field of manufacturing science, the ability to change has been identified as a crucial issue in the recent years. With a holistic vision for the future of manufacturing science, the report [START_REF] Manufuture | Report of the European DG Research, European Conference Manufuture'2003, European Manufacturing of the Future[END_REF] emphasises the need for agility and management of organisational transformations, with manufacturing businesses having "to make continuous reassessments of their core strengths and competencies". Reconfigurable manufacturing systems [START_REF] Harrison | Distributed engineering of modular reconfigurable automation systems[END_REF] enhance the responsiveness to customer requirements. Agile manufacturing aims at providing rapid change over between manufacturing processes [START_REF] Fujii | Agile manufacturing system with autonomously operated square arrayed machining cells[END_REF]. However, such adaptability of technological systems will be fully efficient if they are complemented by the agility of information systems. Furthermore, agility requires the ability of managers to ensure the coherence of their strategy with industrial constraints at the tactical and operational levels of organisation.

From this perspective, the paper focuses on the management of information system transformations. Information system (I.S.) changes are directly induced and constrained by changes in the manufacturing firms' strategies, technologies, or organisation of processes. This issue is especially relevant in the microelectronic domain, which has to adapt competitively to highly reactive markets. Indeed, these companies are subjected to a fluctuating worldwide demand, and must respect constraints generated by high levels of both process and information technologies. This research is the result of the collaboration between the Ecole des Mines de Saint Etienne and the firm STMicroelectronics, resulting in the PhD thesis [START_REF] Chapron | L'urbanisme organisationel : méthode et aide à la décision pour piloter l'évolution du système d'information de l'entreprise[END_REF].

Herein, a process-oriented approach is used to present new tools for change management applied to information systems, which is called "Organisational Urbanism of Information Systems" † . The objective is to take into account organisational factors when one has to decide on transforming the information system, thus not restraining the decision scope by considering only technical factors. To implement this approach it is necessary to analyse rigorously and then formalise the dependency links between processes, which could have an impact on organisational changes.

In section 2, the scientific positioning of the paper is explained and the objectives of the method are defined. The notion of "Organisational Urbanism" is also presented. Then, at the heart of the article, section 3 presents three complementary dimensions that have been chosen for process dependency and the measures of dependency are detailed. The selection of dependency dimensions is based both on the expertise of the industrial partners and on a state of the art of scientific literature. Section 4 focuses on the use of dependency links through a clustering method. Finally, section 5 applies this approach to a STMicroelectronics case study, and outlines the possibilities of organisational diagnosis using the clustering results.

Scientific positioning and objectives of the « Organisational Urbanism"

From Information System Urbanism towards Organisational Urbanism

Few formal approaches are available to answer, with a rigorous and scientific method, the joint transformation of the organisations and their information systems. The first contributions in this perspective are to be found in the field of enterprise modelling: in the domain of automatic control, [START_REF] Malhéné | Gestion des processus d'évolution des systèmes industriels : conduite et méthode[END_REF] represents the evolution trajectory of † Please note that "Urbanism" is used in the paper with the meaning of "City planning". a firm ; in the field of enterprise engineering the concept of "organisation life-cycle" aims at managing enterprise transformations [START_REF] Williams | A handbook on master planning and implementation for enterprise integration programs[END_REF], [START_REF] Geram | Generalised Enterprise Reference Architecture and Methodology[END_REF] ; this idea was also recently expanded by innovative research on virtual firms [START_REF] Camarinha-Matos | Virtual Enterprises: Life cycle supporting tools and technologies, Handbook of Life Cycle Engineering: Concepts, Tools and Techniques[END_REF], [START_REF] Strader | Information infrastructure for electronic virtual organization management[END_REF], (Hoffman and Schlosser, 2001). However, these approaches only provide a partial answer concerning organisational flexibility [START_REF] Dursun | Towards a truly integrated enterprise modelling and analysis environment[END_REF].

In the field of Information System Science, the industrial approach of "Information Systems Urbanism" has been recently developed. Urbanism has the objective to make an existing information system more adaptable to the various types of transformations which can occur within an organisation. The current urbanism methodology was essentially structured via the implementation of industrial experiences (Le [START_REF] Roux | Urbanisation et Modernisation du SI[END_REF], [START_REF] Club-Urba | Pratique de l'Urbanisme des systèmes d'information en entreprise[END_REF], [START_REF] Papadacci | NENO: une approche d'aide à l'arbitrage par l'évaluation qualitative et quantitative de la valeur métier du SI durant la phase amont du processus[END_REF], [START_REF] Chelli | Urbaniser l'entreprise et son système d'Information. Guide des entreprises agiles[END_REF]. This methodology presents the basic concepts necessary to implement information system adaptability [START_REF] Longépé | Le projet d'urbanisation du système d'information[END_REF], [START_REF] Carvalho | Evolution du Système d'Information fondée sur l'Urbanisation -Application au contexte hospitalier[END_REF]. The urbanism procedure has been structured into four well-differentiated levels, to ensure the consistence between the strategy, internal organisation, and information technologies of a firm.

This paper aims at expanding the current urbanism approach by better integrating organisational factors. Indeed, current methods of Information System Urbanism lack scientific tools (i) to manage the organisational view, and (ii) to analyse objectively the consistence between the organisational and technological levels [START_REF] Chelli | Urbaniser l'entreprise et son système d'Information. Guide des entreprises agiles[END_REF].

From this perspective, the overall information system needs to be differentiated from its [START_REF] Vernadat | Techniques de Modélisation en Entreprise: applications aux processus opérationnels[END_REF]), the information system involves not only a technological but also an organisational dimension. To refer to the information system independently of the technology, the term "information processes" will designate information operations without considering whether or not they are technologically implemented. Besides, the "computerised system" represents the technological sub-system. The term "computerised information processes" will represent automated data treatments.

The authors' contribution to the field of urbanism consists in implementing new urbanism mechanisms and tools at the organisational level. This approach has been called "Organisational Urbanism", and focuses notably on the following objectives:

-Defining a modular and generic framework to provide managers with a decisionsupport to manage information system transformations. The organisational urbanism of business processes will be based on criteria clearly distinct from technological constraints.

-Analysing scientifically, the consistence between organisational and technological urbanism. This issue has been identified as a key point in current urbanism methodologies [START_REF] Longépé | Le projet d'urbanisation du système d'information[END_REF], [START_REF] Chelli | Urbaniser l'entreprise et son système d'Information. Guide des entreprises agiles[END_REF].

Based on a process-oriented characterisation of the information system, Organisational Urbanism intends to offer a new view on the organisation and ways to transform it. As we will explain hereafter, the approach is based on the identification of information process clusters, with three main goals: (i) generate relevant clusters of processes, For a complete vision of organisational urbanism, the reader can refer to [START_REF] Chapron | L'urbanisme organisationel : méthode et aide à la décision pour piloter l'évolution du système d'information de l'entreprise[END_REF]. This paper only focuses on the clustering approach, which is the heart of the method, and the direct use of clusters for organisational diagnosis. Organisational Urbanism requires generating clusters which will later constitute relevant organisational units to manage changes. The primary objective is to manage together and within the boarders of a given cluster, most of the changes which concern each process. Each cluster gathers highly interdependent information processes, and thus constitutes a basic and consistent enterprise entity for the deployment of changes. The clustering criteria will be defined thanks to the study of information process dependencies. Therefore, carefully defining the dependency links used to generate process clusters is important, as emphasised in the following sections.

Scientific literature on process dependencies

Following [START_REF] Malone | The interdisciplinary Study of Coordination[END_REF]Crownston, 1994) and(Crownston, 1997), who state that enterprise coordination can be managed through process and entity inter-dependencies, our research is based on the hypothesis that the coordination and then the transformation of the information system can be managed through the direct and indirect interactions between processes. Indeed, every coordination mechanism or every interaction between processes either results from or induces interdependency links. (Hee-Wong, 2000) or [START_REF] Pollalis | Patterns of co-alignment in information-intensive organisations: business performance through integration strategies[END_REF] also confirm that, to manage the evolution of processes, it is necessary to identify the trans-organisational processes, rationalise the flows and supervise their dependencies.

In their coordination theory Crowston and Malone define the coordination as the management of interdependencies among activities or processes (Malone and Crownston, 1994), [START_REF] Crowston | A coordination theory approach to organisational process design[END_REF] . Coordination mechanisms are considered as key components of organisational structures and they can be managed through several types of dependencies among tasks and sub-tasks. Generally, the dependencies are considered at the level of the tasks, and always linked to shared-resources.

Coordination has also been deeply studied in the context of product design and concurrent engineering. In this perspective, (Kusiak and Wang, 1993) and [START_REF] Chen | Decomposition of interdependent task group for concurrent engineering[END_REF] propose a re-organisation of product design processes based on task dependencies. Other process-based approaches like Business Process Reengineering (BPR) include an analysis of dependencies between activities [START_REF] Hwang | On the discovery of process models from their instances[END_REF].

All these approaches advocate that to manage the transformation of a system and to ensure coordination in the long run, studying the dependencies among the basic components is necessary. However, the link between dependencies and the different types of "resources" used by the activities or processes has not yet been detailed. dependencies to implement automated data exchanges among applications. [START_REF] Attie | Specifying and Enforcing Intertask Dependencies[END_REF], [START_REF] Carter | Customizing Internal Activity Behaviour for Flexible Process Enforcement[END_REF] have analysed task dependencies by taking into account information flows among the resources used for each task. [START_REF] Lim | ECEM : an event correlation based event manager for I/O-intensive application[END_REF] uses event dependencies to improve coordination among the sub-components of an information system. [START_REF] Albino | Analysis of information flows to enhance the coordination of production processes[END_REF] go a step further, with the identification of dependencies among pieces of information manipulated by the tasks, in order to detect critical points of coordination among the processes. Based on similar studies of information flow dependencies, the integration of applications turns out to be a compelling objective [START_REF] Castano | A methodology and tool environment for process analysis and reengineering[END_REF]. For instance, [START_REF] Lee | Automatic clustering of business processes in business systems planning[END_REF] applies informational dependencies to determine clusters of processes defined by the pieces of information in use.

Complementary approaches consider the dependencies created by interactions at the level of the actors involved in the processes. Actually, human actors can also be considered as shared resources, generating dependencies among processes. Obviously, specific links between the actors also have to be considered. Moreover, human-centred coordination mechanisms are necessary to ensure work efficiency. In their sociotechnical approach, [START_REF] Yu | Modelling Organisational Issues for Enterprise Integration[END_REF] show that actors of a firm depend on each other: such dependencies must be managed in order to be more reactive in change situations. [START_REF] Dahlgren | Managing inter-firm industrial projects on pacing and matching hierarchies[END_REF] study the impact of the structure of the actors on the output of a firm's processes. In the field of information systems [START_REF] Castro | Towards requirements-driven information systems engineering: the Tropos project[END_REF] also use actor dependencies for a better definition of information system All these contributions concerning task, activity or process dependencies underline several types of dependency links related to (i) technological resources, (ii) shared information, or (iii) human actors, when considering activities within their sociotechnical environment. Based on these conclusions, section 3 specifies the dependency dimensions considered in this approach.

Formalisation of process dependencies

Selection of dependency dimensions

The dependency criteria needed for this research must comply with the change management objective. The goal is to generate clusters of processes whose level of interdependence justifies managing internal changes altogether. Furthermore, since change is dealt with at the organisational level, organisational dimensions have to be considered. Both the scientific state of the art provided by the previous section and a study in the field done by STMicroelectronics has led to the selection of the three following dimensions of dependency: shared technological resources, shared information, or actor relationships. As a matter of fact, the "Organisational Urbanism" method was first designed to fulfil the specific requirements and needs of the company STMicroelectronics. Based on the expertise of information system specialists in the company, each of the three dimensions above has been validated as essential to answer Therefore, there is a need to develop an evaluation of these main dependencies, using basic measures of more elementary dependency links.

Moreover, process dependencies will be defined on two levels. Following Malone's coordination theory, most of the current dependency approaches only consider direct dependency among tasks or activities. Organisational Urbanism does consider the direct dependency, at the first stage. This dependency is induced by the sharing of the three "dependency-objects" † corresponding to the three selected dimensions: shared actors, technological resources, or information. But in addition, at the second stage, indirect dependencies are considered: dependencies among the dependency-objects themselves, even when these objects are not directly shared by the processes ‡ . Thus, in the following sections, dependency measures will be formalised covering both † Only necessary to simplify these explanations, the 3 following generic notions, "actors", "technological resources" and "pieces of information", will be named "dependency-objects".

‡ For instance, there is an indirect dependency between two processes when one actor of the first process has a hierarchical link with an actor of the second. Since there is no shared actor between the two processes, there is no direct actor dependency, but an indirect dependency. the three dimensions selected and the direct and indirect dependencies. The three dependency dimensions also define three complementary axes of action to improve coordination. Indeed, the identification of dependencies can also detect constraints and obstacles that confront any system change.

Formalisation of the dependency concept

In the context of change management, two objects are dependent when the modification of one has an impact on the functioning or the transformation of the other [START_REF] Chapron | L'urbanisme organisationel : méthode et aide à la décision pour piloter l'évolution du système d'information de l'entreprise[END_REF]. Given two processes or sets of processes C 1 and C 2 , the goal is to evaluate their dependency dep(C 1 , C 2 ), using a real number in the interval [0,1] † . However the method only requires modelling mutual dependency and not causal dependency. The following restrictive hypothesis can be made: "The dependency between processes is modelled by elementary dependency-links which are neither quantified, nor oriented". This hypothesis implies that no causal information on the dependency link is used, and that the quantification of the dependency will depend only on the number of elementary dependency links, and not on their intensity. The first justification is that, for the purpose of change management, generating clusters of processes presenting strong mutual dependencies is required, and this mutual dependency is independent of the orientation of the causal link. Secondly, a reliable quantification of each elementary dependency links would require a very precise level of detail for the process models.

Considering the large number of dependency links, such a detailed modelling would † In the interval [0,1], 0 means no dependency at all and 1 means full dependency. make the method non applicable to the industrial context. As a consequence of the hypothesis, the mutual dependency is considered here as a restrictive version of dependency † , which fits with the property of symmetry dep(C 1 , C 2 ) = dep(C 2, C 1 ).

In the three following sections, the formalisation and evaluation of the main dependencies actor D A , resource D RE , and information D I will be presented. For each of them, all the elementary dependency links will be identified. These elementary dependency links are not quantified: they are only valuated 0 or 1 depending on their existence (as a consequence of the preceding hypothesis). Then section 3.4 will explain the aggregation of the elementary dependencies to provide the final evaluation of D A , D RE , and DI.

Actor dependency -D A

The actor dependency only concerns human actors, while the dependencies among technological resources are represented with D RE . To formalise D A , the three elementary dependency links are defined: (a) actor sharing, (b) structural dependency and (c) role dependency.

Actor sharing: two processes P n and P m are dependent in terms of actor sharing when they have in common the same person. For each pair of actors A i , A j mobilised, respectively by the processes P n and P m , the function expressing if the actors A i and A j are the same is noted d a (A i , A j ) (Binary value 1, if the actors are identical, otherwise 0). † Further research could also develop a formalisation of causal dependency using non-symmetric measures. The value of d a is obtained for each combination of pairs of actors mobilised by P n and P m . The aggregation function used to define the final dependency between the processes will be specified in the section 3.4.

Structural dependency: two processes P n and P m are dependent when they involve distinct actors who are hierarchically linked in the enterprise organisational chart. The structural dependency between two actors is noted as d s (A i , A j ). d s is measured in reference to the enterprise organisational chart. Conventionally, depth(A i ) is the depth of the node A i in the hierarchy (i.e. the number of edges on the path from the root of the tree to the node A i ). Given any two actors, the Lowest Common Ancestor -LCA(A i ,A j )is the node of greatest depth that is an ancestor of both actors A i and A j .

If Max (Depth(A 1 ), Depth (A 2 )) -Depth(LCA(A 1 ,A 2 )) ≤ α then d s (A 1 ,A 2 )=1 else d s (A 1 ,A 2 )=0.
The variable α represents the number of edges in the organisation chart separating two actors from their Lowest Common Ancestor, from wherein the two actors are no longer considered dependent. This formula can be adapted to the individual structure of an enterprise (depending on the number of hierarchy levels).

Role dependency: two processes P n and P m are dependent when they involve exactly the same role † . For each pair of actors A i , A j mobilised by the processes, the function expressing if the actors A i and A j perform the same role is noted d r (A i , A j ) (Binary value, 1 if the roles are the same, otherwise 0). The value of d r is obtained for each combination of pairs of actors mobilised by P n and P m . † Here, we do not identify similar roles but only identical ones. This level of precision is sufficient with regards to the constraints of the case study, as underlined in the results (section 5). Thus, the notion of role applied to human actors is used. Referring to a state of the art on this subject -e.g. [START_REF] Ould | Business processes: modelling and analysis for reengineering and improvement[END_REF], (Archimate Project, 2003), (ISO 19440, 2005)-, the concept of "role" has been defined in [START_REF] Chapron | L'urbanisme organisationel : méthode et aide à la décision pour piloter l'évolution du système d'information de l'entreprise[END_REF] as the mission and scope of responsibility of an actor, in a specific context. The role is a generic concept used to describe business activity domains by consistent sets of missions and competencies required. Considering change management, the role concept highlights the domains under common responsibility and, therefore, reveals processes which depend greatly on actor competency.

Technological Resource dependency -D RE

Since this study only focuses on information processes, the technological resources involved in the processes are computer applications. The link between processes and hardware is no longer considered as a constraint at the process level. The high performance of the hardware allows most types of process implementation. Thus, two types of resource dependency will be considered:

Software sharing: two processes P n and P m are dependent when they have in common the same software. For each pair of applications S i , S j used respectively by the processes P n and P m , the function expressing if the applications are the same or not, is noted d so (S i , S j ), taking a binary value, 1 if identical, 0 otherwise. The aggregation defining the final dependency between the processes will be specified in the section 3.4.

Data flow dependency: two processes using different applications are dependent if there are significant data flows exchanges between these applications. For each pair of applications, S i , S j used respectively by the processes P n and P m , the data flow dependency is noted d f (S i , S j ).

d f (S i , S j ) can be measured using the outputs of the standard Information System Urbanism method (not the "Organisational" Urbanism). Regardless of enterprise processes, the Information System Urbanism generates software clusters based on the data flow intensity between the applications. These software clusters will be used here to define the data flow dependency. Two applications S i , S j belonging to the same cluster will have a dependency of 1 (full dependency), 0 otherwise † .

Information dependency -D I

The information dependency will be based on the concept of business object, defined in [START_REF] Chapron | L'urbanisme organisationel : méthode et aide à la décision pour piloter l'évolution du système d'information de l'entreprise[END_REF]) as a set of information manipulated by information processes and, whose meaning is unique and shared by all the business actors irrespective of the process. The identification of the business objects in use, and the specification of their boundaries require a contextual expertise of the firm. Such expertise is also needed to model the processes and their boundaries, as underlined in section 5.

The use of the business object concept takes into account the semantics of the information used by the processes regardless of IT implementation, operational data structure and data life cycle. Thus, information dependency is complementary to data flow dependency which only focuses on IT implementation constraints and data structures. Information dependency is evaluated with two elementary dependency links:

Business object sharing: two processes P n and P m are dependent if they share a common † Here again a more precise measure would not have provided the method with any real added-value. The event dependency: two distinct processes are dependent if a business object they share induces a logical coordination between these two processes or, in other words, when their logical sequencing is constrained. This logical coordination covers synchronisation constraints, in the case when synchronism is required at the logical level, and not only induced by an implementation constraint. This dependency is noted d ev (O i , O j ) (Binary value, 2 if the business object is a logical coordination between the processes, 1 otherwise).

From a quantitative point of view, the event dependency will be introduced as one of the factors necessary to calculate business object sharing dependency. When a business object induces a logical coordination between two processes, this information is taken into account to modify the evaluation of the business sharing dependency. This is obtained with the aggregation function (weighting the business objects) -see §3.4.

The calculation of these elementary dependency links related to D A , D RE and D I is summarised in Table 1. Note that the elementary dependency measures are relative to the dependency-objects and thus they have to be aggregated to provide an evaluation of dependency links at the process granularity level. The following section will explain the [Insert Table 1 about here]

Aggregation of dependency measures

The The weight coefficients (w 1 , w 2 , w 3 , w 4 , w 5 ) chosen for this aggregation can be fine tuned depending on the targeted diagnosis.

Figure 1 sumarises the different steps of the dependency measure computation described above. The computation of the three main dependencies produces a process graph where the nodes represent the processes and the edges represent the dependency between the processes. This graph will be represented as a matrix and analysed to identify relevant clusters, as explained in the following section.

[Insert figure 1 about here]

Clustering of processes

Our approach will now use a clustering method in order to generate a representation of the Information System based on consistent and highly dependent sets of processes, Figure 2 presents an overview of the decision-support procedure. First, process mapsin their technological and organisational environment -are generated. These enterprise models contain all the company data required for the proceeding steps. These models are built upon a meta-model specified in [START_REF] Chapron | Managing the information system evolution: the microelectronics case study[END_REF]. Data collected at this stage is used to evaluate the process dependencies, according to procedures in section 3.

This step generates a process dependency graph. Then, mathematic clustering is applied to this dependency graph. The goal is to extract groups of processes which are coherent with change management criteria. Cluster maps provide concise and relevant information in order to supply decision-making supports.

[Insert figure 2 about here]

Selection of a clustering approach

Clustering consists in grouping objects, evaluated on a matrix of distance (or dissimilarity) between each pair of objects, into a limited number of homogeneous subsets. Clustering reveals a structure in a collection of objects. There is a large number of clustering approaches, mainly based on mathematical theories of graph analysis (Jain et al.,99) where sub-clusters can be contained in larger clusters. But, the distance between each pair of objects or object clusters has to be known, in order to build a partition.

The objective of the method here is to obtain distinct clusters of processes, considered as consistent organisational units to achieve change goals: all the processes within a

given cluster have to be managed together when there is a situation of change. This objective induces the use of a hard, non incremental, and determinist method. Moreover, the measure of distance between two processes (i.e. dependency measure) is a nonoriented and aggregated distance (which means it is a unique criterion; two nodes of the graph are only connected with one non-oriented edge). The size of the data computed is quite small (even when considering hundreds of processes) compared to sophisticated algorithms specialised in the treatment of large databases. These criteria put forth the classification methods.

Two approaches characterise most of the classification methods: agglomerative algorithms (bottom-up) and divisive algorithms (top-down). Divisive methods have rarely been applied, especially because they consume more calculation-time than the agglomerative algorithm. Therefore, the Agglomerative Hierarchical Classification (AHC) was selected.

The clustering procedure

The dependencies defined in section 3 have to be calculated for each pair of processes considered, and result in a dependency matrix providing the inputs for mathematical Then, before applying the AHC algorithm, a multi-dimensional scaling method (MDS) is used. The goal is to detect meaningful underlying dimensions that explain the observed distance between the investigated objects. In the case of this method, the MDS plays the role of pre-treatment, like a filter while preserving the useful information. The result is a better partition, by creating more homogeneous clusters when applying the classification method [START_REF] Lebart | Statistique exploratoire multidimensionnelle[END_REF].

Thirdly, the AHP method is finally applied. The principle of the classification algorithm is to create a partition at each step. This partition is obtained by aggregating the nearest elements two by two. What are called "elements" can be the objects to classify as well as the clusters generated by the algorithm [START_REF] Berkhin | Survey of clustering data mining techniques[END_REF]. A partition is a set of clusters defined at a certain level in the tree. The algorithm does not only give a partition of the elements but a partition hierarchy which is usually called a dendrogram (see Figure 3). The dendogram is then cut to get the final partition, which will be represented as a cluster map.

[insert here figure 3]

The mathematical criteria used to position the final "cut" of the dendogram has to be precisely defined. Here, the partitions are made to minimise the information loss associated with each cluster. In this objective, Ward's hierarchical method was used [START_REF] Ward | Hierarchical grouping to optimize an objectimize function[END_REF]. Two clusters are aggregated in such way that the inter-class inertia is the least, so that the classes remain homogeneous. More details on algorithms and mathematic criteria in use can be found in [START_REF] Chapron | L'urbanisme organisationel : méthode et aide à la décision pour piloter l'évolution du système d'information de l'entreprise[END_REF].

Several cluster maps

As explained previously, three main dependencies between processes are considered.

Each dependency (actor, resource, information) can be used in the clustering method to provide a specific cluster map. Furthermore, these three types of dependencies can also be aggregated in a global dependency which supplies a cluster map of the aggregated dependency between the processes of the enterprise.

Thus the approach can produce four different maps, either applied to the existing or targeted information system. Which map will be used depends on the point of view of the manager. Each map can be used in the strategic and/or tactic approaches of the Organisational Urbanism methodology (see 2.2). These maps can also be compared one with another to highlight internal consistence problems, among the distinct dependencies. This will be illustrated in section 5. The added-value as decision-support tool is situated at the pragmatic level: [START_REF] Williams | A general theory of classificatory sorting strategies -Hierarchical system[END_REF] explains that it is not a question of reaching a true or false result, whether probable or improbable, but only profitable or non-profitable.

The case study -ST Microelectronics Company

Context and objectives of the case study

This approach was applied to the Franco-Italian electronic chip manufacturing company First, the rapid evolution of semi-conductor technologies requires constant and heavy investment for a short product life cycle. Second, market fluctuations with 5 to 7 year cycles, make it difficult to forecast the production workload. Microelectronics is a highly specialised industry where IT tools are essential. With the manufacturing process of a wafer requiring more than 300 steps and production units of several thousand employees, the underlying information system has a very high degree of complexity.

This complexity makes almost impossible the analysis and management of information systems without decision-support tools for change management.

On the operational level, the objective is to build a decision-support tool where the precise dependency and clustering calculations are automated and thus non-apparent to the end-user. The successive steps of calculation are embedded in the tool, which improves user-friendliness. The responsibility of the user is to input correct data on the enterprise and its processes. A meta-model specifying all the enterprise modelling concepts required to implement the approach was proposed in [START_REF] Chapron | Managing the information system evolution: the microelectronics case study[END_REF]. The This is a feasibility study. The objective was to test and apply the method on a limited number of processes, in order to test and validate the applicability of the different steps of the method. The experimental case presented gathers fifteen distinct processes. This first industrial test trial will illustrate the generation of cluster maps (section 5.2), as well as some Information System diagnoses that can be deduced (section 5.3). The company is currently deploying the method to an even larger portion of the firm.

Dependencies computation and clustering sequence

Table 2 presents the fifteen processes concerned.

[Insert table 2 about here]

The data collected on these processes is sufficient to obtain the cluster maps. In the following sections, the various steps which generate these maps will be described.

However, the data presented are anonymous to guarantee the enterprise confidentiality.

Dependency calculation steps

In this section, the procedure to calculate the actor elementary dependency is explained, using the example of two processes: P 2 -Process Change Review Board and P 7 -Scrap Management. Of course this procedure must be repeated for each additional pair of processes, as well as for the two other main dependencies: resource and information dependencies. The company organisation chart is used to identify the depth of the Lowest Common Ancestor for each pair of actors. Table 3 shows the calculation results for the structural dependency applied to each pair of actors. The structure of the organisational chart has been simplified for the case study. In this context, the value of the variable α is 4 (see 3.4), which means that two actors separated by more than four hierarchical levels are not considered as dependent.

[Insert table 3 about here]

The aggregation function (see section 3.5) applied to the actor structural dependency matrix provides the value 0.44.

b. Actor role dependency -d R

The matrix of Table 4 identifies the different roles for the processes P 2 and P 7 .

[Insert table 4 about here]

The aggregation function applied to the actor role dependency matrix gives the value 0.37.

c. Actor sharing -d A

The two processes P 2 and P 7 share two actors -The Device Engineer and the Process Engineer (see Table 3). The calculation procedure is the same as for the role dependency. The actors of the two processes are compared in a matrix, later aggregated using the same aggregation function as for role dependency (section 3.5). d A takes the value 0.25. The actor main dependency aggregates the elementary dependencies using the following formula:

The coefficients of aggregation applied to the elementary dependencies must be defined depending on the objectives of the analysis. In this study, the roles are highlighted (high coefficient) because they represent functional dependencies between the processes.

Such dependencies are considered by the manager as crucial in situations of evolution.

Cluster maps building

The previous dependencies are calculated for the three main dependencies (actor -D A , resource -D R and information -D I ) and for all the processes. Table 5 represents the actor dependencies for all the processes studied:

[Insert table 5 about here]

This matrix provides the input for the mathematical clustering analysis. To execute the multi-dimensional scaling method and the agglomerative hierarchical classification, the software XLStat, developed by the company Addinsoft, was used. First, the multidimensional scaling method was applied to improve the quality of the data. Then the agglomerative hierarchical classification (AHC) was employed. In the AHC method, the calculation of the aggregation criterion is based on the Ward method. This approach calculates the loss of inertia for each new group composition. The cut on the dendrogram must be placed when the inertial loss is the most important. represents the results of the analysis for the actor as well as resource dependencies.

[Insert figure 4 about here]

Diagnosis applied to the case study

At this stage of the method, the initial goal has been reached: the generation of cluster maps. However, the important point is to facilitate the use of such maps for decisionsupports applied to information systems diagnosis and transformation management. In [START_REF] Chapron | L'urbanisme organisationel : méthode et aide à la décision pour piloter l'évolution du système d'information de l'entreprise[END_REF] several levels of decision-support have been developed. However, it is not possible to present all of them. Therefore, in section 5.3.1, only one example of diagnosis will be discussed (the analysis of consistence between distinct cluster maps).

A brief overview of other decision-support systems will be given in section 5.3.2.

Consistence among the three cluster maps

The clustering sequence has generated a map for each of the three dependency dimensions. But, as shown in Figure 4 for the case study, these cluster maps are rather different. The Information System diagnosis presented below consists in comparing these three maps and identifying potential inconsistencies. The comparison can be automated. The potential inconsistencies have to be further analysed with contextual information from the company, to confirm if they point out (or not) any organisational weaknesses, which could later lead to remedial actions. An example of this diagnosis is presented via STMicroelectronics case study.

First, a potential organisational weakness can occur when different processes belong to the same cluster in both the actor and resource maps, but not in the information one. These processes are dependent because of the actors and the software in use, but with a low level of information sharing. This situation has to be further analysed to check if there is a lack of information sharing or technology deployment for the processes. In the case study, this situation was identified for processes P 2 , P 7 , P 8 . A contextual analysis by STMicroelectronics has confirmed the necessity to improve information technology: two major projects for processes P 2 and P 8 have been launched.

A second potential inconsistence appears when several processes belong to the same cluster in the resource map, but to different clusters in the information and actor maps. This situation could point out a lack of cohesion between the information systems and the actor/information organisation: the same software answers the needs corresponding to several of the enterprise's sectors. Sometimes, using different applications or specifically configured software can lead to performance gains. However, in the case of STMicroelectronics, this situation occurs with several processes because the Manufacturing Execution System, at the heart of the manufacturing information system, is transversal to a lot of processes. Therefore, this situation has been justified by the context and there is no inconsistence here.

A third example of potential inconsistence occurs when several processes belong to the same clusters in the actors and information maps, but not in the resource map. The process actors are interdependent from each other and share information, but they do not use common software. This situation can emphasise a need of information technology at the interface between several processes. In the case study, this situation was identified for the pairs of processes {P 11 ; P 12 } and {P 9 ; P 10 }. However, in both cases, a software at the interface of the processes already existed, but the processes were not in the same resource cluster because of other dependency links considered during the clustering.

Thus, this situation is not inconsistent, and outlines the limits of the clustering support.

These three examples show the importance of the contextual factors. Each of them has led to a different conclusion: (i) confirmation of inconsistence, (ii) rejection because the situation is accepted, and (iii) rejection because the results of the clustering approach are not sufficient. Via these examples, three types of inconsistence have been put forward.

However, other types of inconsistencies have been identified in [START_REF] Chapron | L'urbanisme organisationel : méthode et aide à la décision pour piloter l'évolution du système d'information de l'entreprise[END_REF], and their diagnosis has been systematised. A generic grid of most potential inconsistencies has been proposed. Each inconsistence corresponds to a given gap among the three cluster maps. The clusters are represented in a matrix so as to calculate these gaps.

Then, the generic grid is applied to signal potential inconsistencies. Of course the contextual analysis remains necessary to confirm these alarms.

Complementary diagnoses: perspectives

Other diagnoses, based on the use of the cluster maps, have been proposed in [START_REF] Chapron | L'urbanisme organisationel : méthode et aide à la décision pour piloter l'évolution du système d'information de l'entreprise[END_REF]. They are briefly introduced herein to illustrate the added-value of the approach.

At a primary level, some decision-support has been proposed to evaluate the existing system:

-The consistence analysis applied to the three dependency dimensions of the information system, which has been explained in the previous section. -A maturity analysis on the clusters, used to build targets and transformation scenarios for the information system. Referring to the standard Capability Maturity Model (Carnegie Mellon University, 2002), the maturity of the process clusters is assessed. This information is further used to ease management of change.

-An analysis of the strategic weight of the clusters. Based on the evaluation of the contribution of each process to the strategy of the firm, the overall strategic weight of the clusters is calculated. This indicator complements the maturity level, in helping manage IT evolutions.

At another level, some complementary decision-support focuses on the comparison of the alternative solutions for the information system transformations:

-An analysis of the quality of the cluster maps. This analysis is based on the definition of three quality indicators: the level of separation between clusters of a same map, the internal degree of cohesion of a cluster, the number of clusters in a map. These indicators can evaluate the quality of the clustering. But overall, they are used to characterise the transformation between two configurations of the information system (pre and post change). same information system, the dependency gaps are identified among the cluster maps from one configuration to the other. Then, the two indicators "extension" and "amplitude" of the transformation are calculated. This information is useful in selecting among several change alternatives.

Conclusion and perspectives

This paper has presented the dependency approach and clustering method used to implement Organisational Urbanism of Information Systems. Based on models formalising knowledge of the business processes and their technical/organisational environment, Organisational Urbanism provides process cluster maps, which present new organisational views. The method tends to be generic. The models were developed for only one enterprise, but offer the necessary flexibility to be adapted to other enterprises and domains.

This method makes use of simplified enterprise representations. Of course, these models cannot integrate all the complexity and exhaustiveness of the existing interactions among the various components of the systems. The definition of the dependencies could thus be refined, by describing more precisely the existing links (for example, the interservices links or the trans-functional co-ordinations in the company organisational chart). Nonetheless, the cluster maps are synthetic decision-support systems that managers can deploy and adapt to their needs. At each level, the different dependency maps provide tools that, when in the hands of the managers, can forecast highlight and The implementation of this methodology on fifteen processes of the company ST-Microelectronics has confirmed the feasibility of the approach. The successive steps of calculation are embedded in the tools which generate the clusters. Meanwhile, the case study has underlined the fact that each implementation of the methodology requires fine tuning to fit with the internal enterprise context and structure. Beyond the feasibility study, the real benefits of the method will appear with an even broader application.

Indeed, Organisational Urbanism should provide a real added-value when the number of processes and interactions considered is high and justifies a systematic and mathematical analysis, as proposed. After the success of the first tests described here, ST-Microelectronics is currently generalising process modelling on several production sites. Furthermore, other decision-support systems using process clusters have been suggested, notably: (i) identifying the inconsistence between two information systems when any company has to be interconnected with an industrial partner, and (ii) managing inter-plants benchmarks, and process transfers from one production plant to another.

F

P5 P4

P5 1 0,07 0,28 0,00 0,09 0,00 0,00 0,00 0,00 0,00 0,44 0,00 0,00 0,00 0,00 P4 0,07 1 0,03 0,00 0,04 0,02 0,01 0,01 0,01 0,06 0,11 0,05 0,01 0,00 0,00 P6 0,28 0,03 1 0,00 0,26 0,03 0,00 0,00 0,02 0,01 0,14 0,00 0,01 0,00 0,00 P14 0,00 0,00 0,00 1 0,00 0,00 0,01 0,18 0,00 0,08 0,00 0,00 0,00 0,00 0,10 P1 0,09 0,04 0,26 0,00 1 0,00 0,00 0,00 0,00 0,00 0,26 0,02 0,00 0,00 0,00 P8 0,00 0,02 0,03 0,00 0,00 1 0,16 0,03 0,15 0,30 0,00 0,17 0,31 0,02 0,10 P11 0,00 0,01 0,00 0,01 0,00 0,16 1 0,03 0,00 0,05 0,00 0,15 0,23 0,01 0,27 P9 0,00 0,01 0,00 0,18 0,00 0,03 0,03 1 0,21 0,24 0,00 0,04 0,19 0,00 0,14 P10 0,00 0,01 0,02 0,00 0,00 0,15 0,00 0,21 1 0,08 0,00 0,02 0,18 0,00 0,00 P2 0,00 0,06 0,01 0,08 0,00 0,30 0,05 0,24 0,08 1 0,00 0,35 0,10 0,00 0,14 P3 0,44 0,11 0,14 0,00 0,26 0,00 0,00 0,00 0,00 0,00 1 0,05 0,00 0,00 0,00 P7 0,00 0,05 0,00 0,00 0,02 0,17 0,15 0,04 0,02 0,35 0,05 1 0,02 0,01 0,02 P13 0,00 0,01 0,01 0,00 0,00 0,31 0,23 0,19 0,18 0,10 0,00 0,02 1 0,02 0,13 P15 0,00 0,00 0,00 0,00 0,00 0,02 0,01 0,00 0,00 0,00 0,00 0,01 0,02 1 0,12 P12 0,00 0,00 0,00 0,10 0,00 0,10 0,27 0,14 0,00 0,14 0,00 0,02 0,13 0,12 1

  system. Referring to the definition provided by

  to manage changes ; (ii) analyse these clusters to define general transformation targets and trajectories ; and (iii) manage future projects in coherence with this overall cluster analysis.

  Resource-based dependencies have been studied in other fields. Notably, the management of information systems has induced research on the automation of information processes. The development of workflows has encouraged studies of tasks

  house needs of change management. As a consequence the three aforementioned dimensions have been implemented to evaluate process dependencies, relative to: -human actor relationships, called Actor Dependency D A ; -use of technological resources, called Resource Dependency D RE ; -information sharing, called Information Dependency D I.

  object. For each pair of business objects O i and O j used by the processes P n and P m , the function expressing if the business objects are the same is noted d o (O i , O j ). The aggregation function used to calculate the final dependency between the processes will be specified in the section 3.4.

  calculate D A , D RE and D I .

n

  and m are the numbers of dependency-objects O i / O j (e.g. "actors") involved in the two processes P 1 and P 2 . The elementary dependency measures d s or d f take binary values 0 / 1, while the process dependencies d S , d R take real values in the interval [0,1].In the case of elementary dependencies d r , d a and d so each dependency-object of one process, say P 1 , can only depend on one dependency-object of the other process P 2 .Therefore the dependency matrices among objects are simplified, and we can use "Business Object Sharing" d o , a different function is used in order to apply the event dependency d ev as a weight to re-enforce d o .For k= s or f, (i,j) and for formula described k = i, so thenOnce the aggregation has been accomplished, to calculate all the process dependencies d S , d R , d A , d SO and d F (and considering all the processes within the cope of the study), a classical weighted mean is used to provide the three main dependencies D A , D R and D I.

  account change management criteria. These sets will constitute basic management units to implement changes. The diagnosis will emphasise the transformation consistence with regards to the existing systems. Transformation consistence should consider both the stability and the flexibility of the system.

  . Like any firm in this domain, STMicroelectronics is faced with frequent technological and organisational upheavals and has strong needs for the joint management of both the transformation of the information systems and the organisational structures. Indeed, this industry is characterised by a twofold challenge.

  enterprise models cover the processes, their organisational context, and technological environment. STMicroelectronics has chosen the software and modelling environment MEGA Process and MEGA Architecture® to implement the Meta-model. All the data the dependencies and generate the clusters are extracted from enterprise models.

  analysis, between the structure of the computer system and the organisational clusters. The objective is notably to evaluate the consistence between organisational and technological urbanism.

-

  An analysis of the transformation effort. Considering two configurations of the

  and the weaknesses of the possible evolutions of the information system.

  Figure 1. Aggregation procedure for the actor dependency Figure 2. Clustering approach Figure 3. Agglomerative hierarchical clustering method Figure 4. Actor and resource dependency dendrograms

  objective here is to aggregate elementary dependency measures applied to the dependency objects (i.e d s , d r , d a , d so and d f ) in order to obtain the corresponding process dependency d S , d R , d A , d SO and d F .First, an aggregation function is applied to d s and d f to calculate d S and d F for two processes P 1 and P 2 .

Table 5 .

 5 Actor dependency matrix for each pair of processes.

	o r
	P
	e
	e r
	R
	e
	v i e
	w
	O n l
	y

P 15 P 12


Marie-Pierre Drogoul-Vey

Catherine Marimoutou

Andrée Robaglia-Schlupp

Mouna Beerli

Jean-Albert Gastaut

Pierre Cau

Isabelle Poizot-Martin

Determinants and evolution of Squamous Intraepithelial Lesions in HIV-infected women, 1991-2004

Keywords: HIV infection, Pap smear, Squamous Intraepithelial Lesion, cervix cancer, HAART HIV infection, Pap smear, SIL, cervix cancer, HAART

HAL is

Introduction HIV-infected women have been described as being at higher risk for squamous intraepithelial lesions (SIL) and cervix cancer since the beginning of the HIV epidemic. This led to the inclusion of cervix cancer in AIDS diseases in the 1992 CDC revised classification. Since that time, gynecologic screening including Pap smears has been strongly recommended as a part of the regular follow-up of HIV-infected women {Delfraissy JF, 2004 #24}.

Previous studies have reported the incidence of SIL as between 8.9 and 11.5/100 person-years [START_REF] Massad | Evolution of cervical abnormalities among women with HIV-1: evidence from surveillance cytology in the women's interagency HIV study[END_REF][START_REF] Schuman | Longitudinal study of cervical squamous intraepithelial lesion in human immunodeficiency virus (HIV)-seropositive and at-risk HIV-seronegative women[END_REF]. Different factors have been reported to be associated with SIL occurrence, principally the existence of Papilloma virus (HPV) infection [START_REF] Cardillo | CD4 T-cell count, viral load, and squamous intraepithelial lesions in women infected with the human immunodeficiency virus[END_REF][START_REF] Massad | Evolution of cervical abnormalities among women with HIV-1: evidence from surveillance cytology in the women's interagency HIV study[END_REF][START_REF] Hawes | Increased risk of high-grade cervical squamous intraepithelial lesions and invasive cervical cancer among African women with human immunodeficiency virus type 1 and 2 infections[END_REF][START_REF] Schuman | Longitudinal study of cervical squamous intraepithelial lesion in human immunodeficiency virus (HIV)-seropositive and at-risk HIV-seronegative women[END_REF][START_REF] Heard | The impact of HIV antiviral therapy on human papillomavirus (HPV) infections and HPV-related diseases[END_REF][START_REF] Harris | Incidence of cervical squamous intraepithelial lesions associated with HIV serostatus, CD4 cell counts, and human papillomavirus test results[END_REF][START_REF] Lillo | Determination of Human Papillomavirus (HPV) load and type in high-grade cervical lesions surgically resected from HIV-Infected women during follow-up of HPV infection[END_REF]) and immunosuppression [START_REF] Cardillo | CD4 T-cell count, viral load, and squamous intraepithelial lesions in women infected with the human immunodeficiency virus[END_REF][START_REF] Schuman | Longitudinal study of cervical squamous intraepithelial lesion in human immunodeficiency virus (HIV)-seropositive and at-risk HIV-seronegative women[END_REF][START_REF] Heard | The impact of HIV antiviral therapy on human papillomavirus (HPV) infections and HPV-related diseases[END_REF][START_REF] Taylor | Genital dysplasia in women infected with human immunodeficiency[END_REF][START_REF] Harris | Incidence of cervical squamous intraepithelial lesions associated with HIV serostatus, CD4 cell counts, and human papillomavirus test results[END_REF]. The role of highly active antiretroviral therapy (HAART) is still debated -some studies have reported a lower incidence of SIL after 1996 but the direct influence of HAART on incidence of SIL is not well defined [START_REF] Heard | The impact of HIV antiviral therapy on human papillomavirus (HPV) infections and HPV-related diseases[END_REF].

The aim of this study was to describe cervical Pap smear results in HIV-infected women followed up since 1991 and to compare women diagnosed with SIL to others in regard of previously identified risk factors (age, mode of contamination, CD4 cell count, antiretroviral treatment).

Results

The data for 423 women screened for cervical SIL between September 1991 and June 2004 were collected. The patients were predominantly contaminated through heterosexual intercourse (48.5%) and intravenous drug use (IDU, 41.1%). When the first Pap smear was performed (baseline), the median age was 33 years (range: 17 to 75 years) with a median exposure to HIV for 5 years. Their median CD4 cell count was 341 cells/mm 3 , 56.5% were antiretroviral treated, 16.1% receiving monotherapy, 14.9% bitherapy and 25.5% tritherapy or more (HAART) (Table 1). Antiretroviral treatment exposure differed according to the time period of first Pap-smear performance, 35.7% of patients were treated before 1995, 48.2% in 1995-1996, and 79.3% after 1996. For the 239 patients who were treated, median antiretroviral exposure was 1.2 years in the first two periods but 3 years in the post-1996 period. Furthermore, the proportion of treated patients increased during follow-up. 2. The 220 patients with SIL were more likely contaminated through intravenous drug use. They were younger, examined for the first time sooner (72.3% were first examined ≤1996 compared to 43.0% in the non SIL group), had a lower CD4 cell count and were antiretroviral treated in about the same proportion but more frequently with mono or bitherapy. Those differences were significant as well when comparing baseline or last characteristics (i.e. at the time of SIL or at the last Pap smear for patients without SIL). It is to be noted that most of patients characteristics were considered only at baseline whether patients presented SIL at this time (70% of the SIL group) or they were seen only once (48% of the non SIL group). However, considering the follow-up characteristics increased the differences (Table 2).

Characteristics of patients with or without SIL

The multivariate logistic regression including characteristics at the time of SIL finally found a higher risk of SIL when patients were younger (OR=0.95, CI=0.92-0.98 for one year younger), examined for the first time ≤1996 (OR: 1.88, CI: 1.12-3.22), had a lower CD4 cell count (OR: 0.98; CI: 0.97-0.99, for each decrease of 10 cells/mm 3 ). Adjusted on all these parameters and compared to no treatment, treatment with HAART tended to be protective for SIL diagnostic (OR: 0.57; CI: 0.31-1.04, p=0.065). No difference was observe for mono or bitherapy. The association between contamination by IDU and SIL diagnosis was no longer significant.

In regard of follow-up data, the median delay between first and second Pap-smear examination was 10.5 months [IQ: 6-18 months]. It did not differ between women with or without SIL neither between those seen first ≤1996 or after.

SIL evolution

Among the 220 women with SIL, 142 had follow-up data after first SIL (65.5% low grade and 34.5% high grade). They were 30.3% (n=43) to present a worsen SIL during follow-up

Cancer of the cervix

In the whole sample of patients, 21 (5.0%) were diagnosed once with cervix cancer, three at first examination and 18 during follow-up, most of them after a previous diagnosis of low or high-grade SIL. Majority (13/21) of these women has been included in the database ≤1996, but only 5 cancer were diagnosed at this period. At inclusion, they were 7/21 classified CDC stage C, 62% were contaminated with UDI (n=13), their median CD4 cell count was 315/mm3 (range: 15-1259). At this time only 3 patients received tritherapy, 4 bitherapy, 6 monotherapy of nucleoside analogues and 8/21 were not treated.

Discussion

In this historic study, the prevalence of SIL was high, more than a half of women presenting SIL at least once, and diagnosis of SIL was predominantly performed at first Pap smear examination. The case control analysis confirms the pejorative role of immuno-suppression on SIL risk. Women with SIL were also younger and more likely included before 1996.

However, even adjusted on theses factors HAART tended be protective of SIL diagnosis.

Moreover, unless patients were somehow regularly screened with Pap smears, Pap smear classification aggravated in 30% of patients with SIL and conducted to cancer in 11%.

This study corresponds to data collected under routine conditions in women followed up in an HIV-specialized unit. However, gynecologic screening was not systematically accepted by the women, who could choose to be followed up by a gynecologist out of the unit or not followed up at all. A selection bias cannot be denied. Nevertheless, the repartition of contamination mode, age and antiretroviral treatment in this sample is similar to that of the whole population of women followed up in the unit.

The association between low CD4 cell count and SIL has largely been reported in literature [START_REF] Cardillo | CD4 T-cell count, viral load, and squamous intraepithelial lesions in women infected with the human immunodeficiency virus[END_REF][START_REF] Massad | Evolution of cervical abnormalities among women with HIV-1: evidence from surveillance cytology in the women's interagency HIV study[END_REF][START_REF] Hawes | Increased risk of high-grade cervical squamous intraepithelial lesions and invasive cervical cancer among African women with human immunodeficiency virus type 1 and 2 infections[END_REF][START_REF] Schuman | Longitudinal study of cervical squamous intraepithelial lesion in human immunodeficiency virus (HIV)-seropositive and at-risk HIV-seronegative women[END_REF][START_REF] Taylor | Genital dysplasia in women infected with human immunodeficiency[END_REF][START_REF] Harris | Incidence of cervical squamous intraepithelial lesions associated with HIV serostatus, CD4 cell counts, and human papillomavirus test results[END_REF], although the significant threshold may differ. For Harris et al., there was no difference in the two-year incidence of SIL between HIV-infected women over 500 CD4/mm 3 and seronegative women, the incidence rose when the CD4 cell count declined <500/mm 3 [START_REF] Harris | Incidence of cervical squamous intraepithelial lesions associated with HIV serostatus, CD4 cell counts, and human papillomavirus test results[END_REF]. However, Massad et al. found that the significant cut-off was 200 CD4 mm 3 [START_REF] Massad | Evolution of cervical abnormalities among women with HIV-1: evidence from surveillance cytology in the women's interagency HIV study[END_REF]. In our study we considered CD4 cell count as a continuous variable, if converted in dummy-variable in three In our study, younger women were at higher risk of developing SIL, suggesting either lower protection against infection due to younger age or more risky behavior for HPV infection. In recent studies focusing on the effects of HAART, age was not reported as a risk factor for SIL [START_REF] Schuman | Longitudinal study of cervical squamous intraepithelial lesion in human immunodeficiency virus (HIV)-seropositive and at-risk HIV-seronegative women[END_REF][START_REF] Ahdieh-Grant | Highly active antiretroviral therapy and cervical squamous intraepithelial lesions in human immunodeficiency virus-positive women[END_REF][START_REF] Heard | The impact of HIV antiviral therapy on human papillomavirus (HPV) infections and HPV-related diseases[END_REF][START_REF] Taylor | Genital dysplasia in women infected with human immunodeficiency[END_REF]). However, patients with cervix cancer were older, suggesting the necessity of longer duration of either HPV or HIV infection.

Patients contaminated though IDU are suspected of being at more at-risk behavior in terms of sexual activity or addiction to tobacco. We did not recorded such behavioral data but no difference between women infected with IDU and others was observed in multivariate analysis. Moreover, Schumann et al. in the HER study did not find any association between SIL and tobacco use or sexual activity [START_REF] Schuman | Longitudinal study of cervical squamous intraepithelial lesion in human immunodeficiency virus (HIV)-seropositive and at-risk HIV-seronegative women[END_REF]).

In the literature, a slight effect of HAART on SIL occurrence or regression has been reported [START_REF] Ahdieh-Grant | Highly active antiretroviral therapy and cervical squamous intraepithelial lesions in human immunodeficiency virus-positive women[END_REF][START_REF] Heard | The impact of HIV antiviral therapy on human papillomavirus (HPV) infections and HPV-related diseases[END_REF][START_REF] Taylor | Genital dysplasia in women infected with human immunodeficiency[END_REF]), but it appeared to be mediated by a CD4 increase [START_REF] Ahdieh-Grant | Highly active antiretroviral therapy and cervical squamous intraepithelial lesions in human immunodeficiency virus-positive women[END_REF].

Our results concluded to a trend in favor of a protective role of HAART compared to no treatment after adjustment on CD4 cell count and year of inclusion. However, HPV is known to interfere with SIL and HAART has no direct effect of on HPV infection [START_REF] Heard | The impact of HIV antiviral therapy on human papillomavirus (HPV) infections and HPV-related diseases[END_REF]).This trend should eventually be confirmed on larger sample with HPV infection recorded. In this study, HPV coinfection was not tested in all of the patients since the study began before HPV detection was routinely performed. However, French guidelines for HIV infection care do not recommend HPV testing but a systematic annual Pap smear if CD4>200/mm 3 and twice a year in case of CD4<200/mm 3 {Delfraissy JF, 2004 #24}. The main result of this study is that unless for cancer occurring on previously observed abnormal cervix, most of abnormalities were detected at the first Pap smear performance. This was particularly true when considering the oldest data (i.e. first Pap ≤ 1996). These patients were less antiretroviral treated or if so with less effective therapy. Moreover, they had a lower CD4 cell count at the first Pap smear. So, it can be concluded that evolution of care after HAART introduction more likely than direct effect of HAART had a major impact on SIL risk in HIV infected patients. However, even with antiretroviral treatment, we observed an aggravation of SIL classification in 30% of women. A previous study by Delmas et al. associating data of 20 women of this cohort showed that 8% of the women with CIN1 had progressed to high-grade lesions at one year [START_REF] Delmas | Cervical squamous intraepithelial lesions in HIVinfected women: prevalence, incidence, and regression. European Study Group on Natural History of HIV Infection in Women[END_REF] and Massad et al observed 14% aggravation of abnormal smears (including ASCUS) at six months [START_REF] Massad | Evolution of cervical abnormalities among women with HIV-1: evidence from surveillance cytology in the women's interagency HIV study[END_REF]. In this study no determinant significantly differed women who progressed from those who did not. No specific role of CD4 cell count was also observed in Massad et al study after adjustment of HPV status [START_REF] Massad | Evolution of cervical abnormalities among women with HIV-1: evidence from surveillance cytology in the women's interagency HIV study[END_REF]).

We also found women diagnosed with SIL at risk of progression to cervix cancer suggesting a lack of either SIL management or efficiency of treatment in this population. The delay between first and second Pap smear did not differ between patients with or without SIL, favoring the hypothesis of problems in curative therapy options in these patients. The high frequency of SIL recurrence is well known in this population even when HAART treated and has been reported elsewhere [START_REF] Gilles | Management and outcome of cervical intraepithelial neoplasia lesions: a study of matched cases according to HIV status[END_REF]. Finally, the incidence of cancer was high in our study compared to results observed in US-HIV-infected women included in a prevention program [START_REF] Massad | Low incidence of invasive cervical cancer among HIV-infected US women in a prevention program[END_REF]). This result seem coherent with the observation of increase of cervix cancer between the 1993-1996 and 1997-1998 periods by Italians [START_REF] Franceschi | Incidence of AIDS-defining cancers after AIDS diagnosis among people with AIDS in Italy, 1986-1998[END_REF] and the results of the Swiss 12 cohort which find this type of cancer persistently 8-fold higher than in seronegative women with no influence of HAART [START_REF] Clifford | Cancer risk in the Swiss HIV Cohort Study: associations with immunodeficiency, smoking, and highly active antiretroviral therapy[END_REF].

In conclusion, our results reflect change in women care after the introduction of HAART but also stress the necessity to maintain a regular gynecologic screening in HIV-infected women.

The role of specific gynecologic treatment on secondary cancer in HAART-treated patients with SIL should be studied prospectively. 1995 [1993-1997] 1997 [1994-2000] 0

F

  Prospective observational cohort of gynecologic follow-up of HIV infected women referred to a single HIV-specialized hospital care unit, the CISIH-Sud of Marseille, France. Since 1991, this hospital unit has proposed to HIV-infected women a gynecologic consultation with a specialist who systematically performed a screening of cervical squamous intraepithelial lesions (SIL) using conventional Pap smears. The results of Pap smears, demographic data, HIV therapy and CD4 cell count were prospectively collected for women who gave their informed consent.All of the Pap smears were collected using a endocervical cytological brush. The specimens were smeared onto one or two slides, fixed immediately, and then sent to the same laboratory. After technical preparation and Papanicolaou staining, each Pap smear was first examined by a cytotechnologist and then evaluated by the same cytopathologist all through the study period. Pap-smear results were classified using the Bethesda classification (Kurman RJ, Malkasian GD Jr et al. 1991)according to the 2001 consensus guidelines (Wright TC, Cox JT et al. 2002), without subdividing the atypical squamous cells (ASC) into 2 categories because our cohort was set up fifteen years ago and the first Pap smears could not be reclassified. In this database of women with Pap smear results we performed a case-control nested study, comparing women presenting at least once a diagnosis of SIL with those free of SIL at all registered Pap smears. They were compared according to their characteristics at first Pap smears performance (i.e. inclusion in the database) and at the time of diagnostic of SIL for those with SIL or last Pap smear results for others. The associations between age, contamination route, year of first pap-smear performance, CDC stage classification of HIV infection, CD4 cell count, antiretroviral treatment and SIL were evaluated using Chi 2 or Kruskall-Wallis tests and logistic regression. Statistical analyses were performed by STATA.

  Among the 423 women who entered the database, 139 (33%) had Pap smear examination only once. Globally, the median number of Pap smears by women was 3 [IQ: 1-6]. In order to increase the power of analysis, the whole sample of Pap smears was considered to identify patients with SIL. Two hundred and twenty women (52%) presented SIL at least once in the database. The first SIL was considered for this analysis. It was diagnosed at the first Pap smear performance in 154 (70%) women and later during follow-up (median delay of occurrence 25 months; Interquartile: 9-41 months) in 66 (30%). The SIL was classified low grade for 64.5% and high grade for 35.5%. Three patients were diagnosed with cervix cancer at first Pap smear and were excluded of the comparison. The 200 women free of SIL at all Pap smears examinations were 96 (48%) seen only once and for the 104 others had their last Pap smear in a median delay of 28 months [11-60 months]. Characteristics of these patients diagnosis of SIL are summarized Table

  delay of 27 months (IQ: 11-46 months), 33.3% among the low grade SIL and 24.5% among the high grade (p=0.28). Among the 31 low grade SIL, 27 reached high grade and 4 cancer. Moreover, 12 high grade SIL were diagnosed cancer during follow-up.Consequently 16 patients (11.3% of patients with SIL and further examination) presented cervix cancer during follow-up in a median delay of 32 months (IQ: 23-66 months). The median duration between SIL diagnosis and last Pap smear for patients who did not aggravate their Pap smear classification was 45 months (IQ: 18-80 months).The patients who worsen their SIL did not differed of others in regards of their characteristics at the time of first SIL or their year of first Pap smear examination, although their median CD4 cell count tended to be lower 274 versus 344, p=0.13. However, at the time of diagnosis of the worsen SIL, the CD4 cell count distribution did not significantly differed (median 268; IQ: 180-425) and the median delta of CD4 was -25 cells/mm 3 [IQ: -114;+36]. Finally, patients with worsen SIL had had 3 Pap smears [IQ: 2-5] recorded in median between the first SIL and the worsen SIL versus 4 Pap smears [IQ: 3-7] for those with no worsen SIL.

  between first Pap examination and cancer diagnosis was 37 months. Unless for the 3 patients diagnosed at first Pap smear, the range distribution of this delay was 4 to 139 months. At the time of cancer diagnosis, women were median aged of 35 years (range: 26-64 years), 25% had a CD4 cell count <200/mm3, 7/21 were at AIDS stage. The great majority (19/21) were ARV treated, 10/21 receiving at least tritherapy and only 2 a monotherapy. Patients diagnosed with cancer had had 4 Pap smears examination in median. Finally, 18 cervix cancers were diagnosed in the 267 women with follow-up data (median follow-up of 4 years), representing a incidence of cancer of 1.2 per 100 persons-years of follow-up.

  , between 200-499 and >500), both threshold <200 and <500 were significant on the risk of SIL (data not shawn).

Table 1 :

 1 Baseline characteristics of women at the time of first Pap smear, n=423

	o r
	P
	e
	e r
	R
	e
	v i e
	w
	O n l
	y

Table 2 :

 2 Characteristics of patients according to SIL diagnosis, N=420

		Patients with SIL	Patients without SIL	p
		N=220	N=200
	Contamination by IDU	99 (45.0%)	71 (35.5%)	0.05
	CDC classification
	Stage A	72 (32.7)	111 (55.5)	<0.0001*
	Stage B	89 (40.5)	50 (25.0)
	Stage C	59 (26.8)	39 (19.5)
	Year of first examination

  IDU: intravenous drug use; IQR: interquartile range * Proportion of stage C tended to be significantly higher in the SIL group (p=0.08) ** Proportion of stage C was significantly higher in the SIL group (p=0.04)

	Age in years	F o r	Baseline Characteristics	.0001
	Median (IQR) CD4 cell count Median (IQR) Antiretroviral therapy None Monotherapy	32 [27-35] 313 [153-457] P e 104 (47.3) 48 (21.8) e r	35 [30-40] 383 [262-549] 81 (40.5) 20 (10.0)	0.0001 0.0001 <0.0001
	Bitherapy HAART			27 (12.3) 41 (18.6) R	35 (17.5) 64 (32.0)
	e Characteristics at SIL diagnosis or last Pap smear free of SIL Median (IQR) Age in years 32 [28-36] 36 [32-43] v i e CDC classification Stage A Stage B 70 (31.8) 87 (39.5) 111 (55.5) 50 (25.0) w	0.0001 <0.0001**
	Stage C CD4 cell count † Median (IQR) Antiretroviral therapy None Monotherapy		63 (28.6) 313 [156-433] 94 (42.7) 41 (18.6)	39 (19.5) 393 [259-557] 59 (29.5) 15 (7.5) O n l <0.0001 0.0001 y
	bitherapy			36 (16.4)	40 (20.0)
	HAART			49 (22.3)	86 (43%)


Jaroslav Kriz
email: kriz@imc.cas.cz

Jiri Dybal

Emanuel Makrlik

Petr Vanura

Jan Lang

Jaroslav Kříž

Jiří Dybal

Emanuel Makrlík

Petr Vaňura

A proton complex of p-tert-butylcalix[4]arene-tetrakis(N,Ndimethylthioacetamide): NMR evidence and probable structure

Keywords: Calixarene complex, protonation, NMR, DFT Calixarene complex, protonation, NMR, DFT

A proton complex of p-tert-butylcalix[4]arenetetrakis(N,N-dimethylthioacetamide): NMR evidence and probable structure

INTRODUCTION

Calixarene-based molecules produced in rich diversity [START_REF] Gutsche | Calixarenes Revisited[END_REF] have received intense attention in the last years. The some calixarenes having four and five member rings bear some resemblance to the natural cyclodextrins [START_REF] Saenger | [END_REF] in as much they posses a cone-like cavity, which is able to act as a host for a number of guest molecules. As such, they find applications as biomimetic studies [START_REF] Gutsche | Calixarenes Revisited[END_REF].

In addition, they can be modified by attaching carbonyl, ester, amide or thioamide groups to their lower rim. Due to their number (usually four or more) and restricted conformation, these groups often serve as efficient coordination site for various cations. Many studies have focused on their binding ability toward metal ions, predominantly alkali and alkaline-earth but also transition and heavy metal cations. [3][START_REF] Arduini | Inclusion Phenom[END_REF][START_REF] Arnaud-Neu | [END_REF][6][7][8][9][10][11] As experimentally proved, coordination of a metal ion to the groups (say carbonyls) at the lower rim exerts a conformation change on the whole calixarene unit, which usually adopts a more cone-like form changing thus subtly its host properties for a guest molecule. The actual conformation change appears to be strongly dependent on the nature of the ion as well as the calixarene itself. The study of ion coordination to modified calixarenes, including non-metallic ions, first of all proton or hydroxonium ion, thus can be of primary interest to supramolecular chemistry.

Up to now, no complex of proton with any calixarene has been reported. We have studied interactions of H 3 O + with calix [START_REF] Arduini | Inclusion Phenom[END_REF]arenes bearing ester, amide and thioamide groups. All of these substances clearly form an equimolecular complex with H 3 O + . However, the dimethylamide-derivative apparently is not available in a sufficiently pure form, the commercial product being partly complexed with Na + ions and thus not suitable for demonstrative purposes. The interaction of the analogue bearing ester groups is somewhat complicated and will be described in a separate study. [18] In this work, we present NMR evidence for a complex of H 3 O + with the thioacetamide derivative known as lead ionophore, namely p-tert-butylcalix [START_REF] Arduini | Inclusion Phenom[END_REF]arene-tetrakis(N,N-dimethylthioacetamide) (1, cf. Scheme 1). On the grounds of 1 H and 13 C NMR spectra and DFT calculations, we suggest its most probable structure.

RESULTS AND DISCUSSION

Similarly as in our previous papers [13,14], hydrogen bis(1,2-dicarbollyl)cobaltate (HDCC) (Ref.12, for structure see Scheme 1) as a proton source, was used in nitrobenzene-d 5 .

A 2.5 mol excess of water to HDCC converts the protons to the hydroxonium ions H 3 O + .

HDCC is well soluble in nitrobenzene, but calixarene 1 did not allow concentrations above In the following NMR spectra, the signal assignment (which was checked by means of 2D COSY-LR and NOESY NMR spectra for 1 H and HSQC and HMBC spectra for 13 C) corresponds to Scheme 1 where the protons have the same number as the carbons they are attached to. Figure 1 shows 1 H NMR spectra of 1 and its 2:1 and 1:1 mol/mol mixtures with HDCC in nitrobenzene-d 5 . There are relative shifts of almost all signals in a similar direction and magnitude as in the case of complexation of metallic ions by calixarene ligands [3][START_REF] Arduini | Inclusion Phenom[END_REF][START_REF] Arnaud-Neu | [END_REF],

5×10 -3 mol/L. C H 3 C CH 3 CH 3 H H C H e H a O H 2 C C=S N H 3 C
evidencing the formation of a complex of 1 with H 3 O + (in Fig. 1B, the signal of proton 13 is broadened and placed under the number 6). Under higher excess of HDCC, no further shift is observed proving thus that the complex is equimolecular and its stability constant is high NMR spectra [15,19]. The average spectral shape is caused by a rapid interconversion between two identical structures (pinched cone). The free energy barrier is usually as low as 9 kcal/mol [19], which makes this motion fairly difficult to be detected by NMR. A significant line broadening due to this process occurs at very low temperatures (around 188 K) when most of solvents become frozen. An evidence of the pinched cone conformation of 1 is also a chemical shift difference of the axial and equatorial bridging methylene protons 9 [15], which is as large as 1.534 ppm.

It is obvious that attachment of hydroxonium or metal cation hinders the pinched cone pinched cone interconversion. In the presence of the fast chemical exchange of hydroxonium cations (on 1H chemical shift time scale), the calix [START_REF] Arduini | Inclusion Phenom[END_REF]arene molecule is allowed to interconvert only in between the periods when the complex is formed. This results in overall decrease of pinched conepinched cone inteŕconversion rate and thus broadening of some of the signals.

When all of 1 is bound to H 3 O + , the calix [START_REF] Arduini | Inclusion Phenom[END_REF]arene part of the molecule is fixed in a C 4 symmetry so that no interconversion takes place and the NMR signals are narrow again. The decrease of the difference of chemical shifts of diastereotopic methylene protons 9 to 1.311 ppm shows that the cavity adopts a flatter, more cone-like form, presumably due to nearer proximity of the lower-rim polar groups interacting with H 3 O + . Despite of the high stability constant of the complex, the hydroxonium cation remains mobile and it samples different orientations within a single ligand averaging thus the whole structure so that there is only one set of signals in Figure 1C.

Somewhat more information about the structure of the complex 1 . H 3 O + can be obtained from 13 C NMR spectra depicted in show up in DEPT spectra but can be found in ordinary 13 C NMR spectra, their shifts being 197.97 and 195.99 ppm in 1 and the complex, respectively. The down-field shifts of carbon signals 4(8) and 5 (7) probably indicate formation of the conical structure that is close to C 4 symmetry. Quite similar 13 C complexation induced shifts were reported by Arduini et al. [START_REF] Arduini | Inclusion Phenom[END_REF] for complex of tetraamide derivative of calix [START_REF] Arduini | Inclusion Phenom[END_REF] In order to confront our experimental findings with theoretical predictions, the equilibrium geometries of ligand 1 and its complex with H 3 O + were calculated at the B3LYP/6-31G(d) level. DFT calculations have been found to offer good results for H-bonded structures [20] although their full reliability in this field was recently criticized [21,22].

However, recommended more rigorous methods like Moeller-Plesset perturbation approach MP2 [21] or modified functionals with wider basis sets [22] are not practicable for such large systems as 1 or its complex. Considering the rather mild deviations apparently produced by the conventional DFT method, we believe that our calculations give a reasonable approximation to our structures.

In Fig. 3, the optimized structure of 1 is given. As it can be seen, the cone is predicted to be distorted in the way suggested by NMR, i.e. two opposite aromatic rings are much more parallel than it would be expected in ligand 1. As explained above, it is in a good agreement with NMR spectra. In Fig. 4, the lowest-energy-level structure obtained by optimization of the 1 . H 3 O + complex is illustrated. In good agreement with NMR, the calixarene part of the considered complex is more cone-like and distinctly more symmetric than free ligand Although the symmetry of the calixarene part is predicted to be fairly near to C 4 (cf. Fig. 4), the strong hydrogen bonds are not symmetrically distributed as it would seem to be so by NMR spectra. The above explanation that the structure is averaged by a fast motion of the H 3 O + ion in the coordination cavity seems to be hampered by a too high energy barrier connected with the necessary breaking of three to four hydrogen bonds. There is a high probability, however, that such barrier either does not exist or is much lower due to the As the ion rotates, some hydrogen bonds are strengthened simultaneously as other are weakened and the energy surface traversed by this motion has shallow ripples rather than high peaks and deep valleys. It is also probable that the rotation can tunnel through these low barriers.

Finally, the calculated stabilization energy of the 1 . H 3 O + complex is 100.4 kcal/mol, which is a rather high value expressing high stability of the complex under study. This again is in agreement with the findings of NMR. The determination of the actual stability constant of the complex 1 . H 3 O + in the organic phase of the two-phase water-nitrobenzene extraction system is underway in our laboratories.

In conclusion, we found that H 3 O + not only binds to 1 but, in doing so, influences its overall conformation forcing its calixarene cavity to adopt a more symmetric and open shape. This surely influences its properties as a host for some guest molecules. One can thus speculate that inclusion interactions of 1 and similar modified calixarenes could be changed by adding protons to the system, which opens interesting possibilities.

EXPERIMENTAL

Materials and samples. Nitrobenzene-d 5 and p-tert-butylcalix [START_REF] Arduini | Inclusion Phenom[END_REF]arene-tetrakis(N,Ndimethylthioacetamide) (1) were purchased from Fluka, Buchs, Switzerland and were used as obtained. Preparation of hydrogen bis(1,2-dicarbollyl) cobaltate (HDCC) was described in Ref. [14]. Briefly, the cesium (CsDCC) analogue was prepared in the Institute of Inorganic Chemistry, Řež, Czech Republic, using the method published by Hawthorne et al. [17]. In order to obtain its hydrogen analogue, HDCC, 0.2 mol/L solution of CsDCC in nitrobenzene was twice shaken with equal volumes of 15% v/v n-propanol in 1 mol/L H 2 SO 4 in distilled water, followed by tenfold equilibrium shaking with equal amounts of 1 mol/L H 2 SO 4 and NMR measurements. 1 H and 13 C NMR spectra were measured at 300.13 and 75.45 MHz frequency, respectively, with an upgraded Bruker Avance DPX300 spectrometer collecting 64 scans for 1 H NMR and 2×10 4 scans for each type of 13 C NMR spectra. In the latter case, DEPT45 sequence was used and spectra with coherence evolution delays adapted for the spin interaction constants J CH 145, 20, 12, and 6 Hz, respectively, were superimposed in the presented 13 C NMR spectra. COSY-LR and NOESY 2D 1 H NMR spectra as well as HSQC and HMBC 1 H-13 C 2D correlation spectra were measured using an inverse-detection zgradient probe taking 1 kpoints and 256 increments in 640 scans. Phase shifted sine-bell weighting function was applied before Fourier transform.

Computational procedures. All calculations were performed using the GAUSSIAN 03 suite of programs [16]. The molecular equilibrium geometries were fully optimized without geometry constrains by using the Becke's three parameter functional of density functional theory with the correlation of Lee-Yang-Parr (B3LYP) and the 6-31G(d) basis set. Several local configurations near the achieved energy minimum were examined. As the renewed optimizations converged to the same molecular geometry, we believe the achieved energy minimum to be the global one.

  carriers, analytical sensors, catalysts and model structures for

  Scheme 1

4 (

 4 well above 10 3 ). At HDCC/1 ratios between 0 and 1 mol/mol, the signals gradually shift and broaden. At the ratio 1:1, the signals are further shifted but narrow again. Evidently, an exchange between free and bound ligand 1 takes place. Its rate is only moderate considering the signal broadening. The corresponding correlation times of exchange τ ex (in ms) calculated from the broadening of the signals and the relative shifts between the free and bound states are given in Fig.1B, while the relative shifts are shown in Figure1C. These values are rather formal, however, considering the complexity of motions discussed below.

Fig. 1 1 H

 1 Fig.11 H NMR spectra of 0.005 mol/L solution of 1 (A) and its 2:1 (B) and 1:1 (C) (mol/mol) mixtures with HDCC. Signal assignment (cf. Scheme 1) in A, approximate exchange correlation times τ ex (in ms) in B, relative chemical shifts (in ppm) in C.

Figure 2 .

 2 In order to overcome the low sensitivity at 5 mmol/L concentration and overshadowing by huge signals of nitrobenzene-d 5 , four types DEPT-LR spectra are combined here. The signals of C=S carbon 11 did not

Fig. 3

 3 Fig. 3 Two projections of the DFT-optimized structure of free 1 (B3LYP/6-31G(d), hydrogen

Fig. 4

 4 Fig.[START_REF] Arduini | Inclusion Phenom[END_REF] Two projections of the DFT-optimized structure of the 1 . H 3 O + complex (B3LYP/6-31G(d), hydrogen atoms omitted except those of H 3 O + ).

  two other sulfur atoms, which form weak hydrogen bonds with the ion.

  distilled water. After separation of the phases, nitrobenzene was removed by distillation. The product was dried under high vacuum to a constant weight for two weeks. For NMR samples, 5×10 -6 mol of 1 was dissolved in a mixture of appropriate amounts of nitrobenzene-d 5 and 0.001 mol/L solution of HDCC in the same solvent.

  :/mc.manuscriptcentral.com/tandf/gsch Email: suprachem@mail.cm.utexas.

  arene with KSCN. The up-field shift of 6 may be attributed to a decreased electron density on the phenoxy oxygen atom due to a hydrogen bond with H 3 O + . At the same time, 1.98 ppm up-field shift of carbon 11 shows a strong hydrogen bond of the ion H 3 O + with a C=S group. As these shifts indicate really strong hydrogen bonds, the most probable structure appears to be that of H 3 O Combined13 C NMR DEPT45 and DEPT-LR spectra of 0.005 mol/L solution of 1 (A) and its 1:1 (mol/mol) mixture with HDCC (B). Signal assignment (cf. Scheme 1) in A, relative chemical shifts (in ppm) in B; in both parts, nb means residual signals of nitrobenzene.

	Fig. 2
		F o r
				P
				e
				e r
						R
						e
							v i e
					nb				w			0.71 -0.20 0.15 -0.21
	-2.04	nb 1.97		0.84 (+nb)		0.56	B	-2.81		HDCC n O l
													y
																	1
						4,8
					nb
	6	3													13 12
				nb
				5,7			A	10									9
		nb															2
	150	145	140	135	130	ppm		80	75	70	65	60	55	50	45	40	35	ppm

+ directly bonded to both phenoxy oxygen and thiocarbonyl sulphur atoms. We have only one signal for each type of carbon including 11, which indicates a C 4 symmetry of the whole molecule; the simplicity of the corresponding proton spectrum points to the same conclusion. As it is hardly possible to have the H 3 O + ion equally bound by hydrogen bonds to all four C=S groups and all four phenoxy oxygen atoms as well we have to assume a rapid motion of H 3 O + within the coordination site averaging the structure in the time window of NMR (see below).

Acknowledgements

. This work was supported by the Academy of Sciences of the Czech Republic, Project T400500402, the Czech Science Foundation, Project 203/04/P168 and the Czech Ministry of Education, Youth and Sports, Projects MSM 4977751303, MSM 6046137307, MS 0021620835.