885 resultados para post-structural theory
Une étude générique du metteur en scène au théâtre:son émergence et son rôle moderne et contemporain
Resumo:
The theatre director (metteur en scene in French) is a relatively new figure in theatre practice. It was not until the I820s that the term 'mise en scene' gained currency. The term 'director' was not in general use until the I880s. The emergence and the role of the director has been considered from a variety of perspectives, either through the history of theatre (Allevy, Jomaron, Sarrazac, Viala, Biet and Triau); the history of directing (Chinoy and Cole, Boll, Veinstein, Roubine); semiotic approaches to directing (Whitmore, Miller, Pavis); the semiotics of performance (De Marinis); generic approaches to the mise en scene (Thomasseau, Banu); post-dramatic approaches to theatre (Lehmann); approaches to performance process and the specifics of rehearsal methodology (Bradby and Williams, Giannachi and Luckhurst, Picon-Vallin, Styan). What the scholarly literature has not done so far is to map the parameters necessarily involved in the directing process, and to incorporate an analysis of the emergence of the theatre director during the modem period and consider its impact on contemporary performance practice. Directing relates primarily to the making of the performance guided by a director, a single figure charged with the authority to make binding artistic decisions. Each director may have her/his own personal approaches to the process of preparation prior to a show. This is exemplified, for example, by the variety of terms now used to describe the role and function of directing, from producer, to facilitator or outside eye. However, it is essential at the outset to make two observations, each of which contributes to a justification for a generic analysis (as opposed to a genetic approach). Firstly, a director does not work alone, and cooperation with others is involved at all stages of the process. Secondly, beyond individual variation, the role of the director remains twofold. The first is to guide the actors (meneur de jeu, directeur d'acteurs, coach); the second is to make a visual representation in the performance space (set designer, stage designer, costume designer, lighting designer, scenographe). The increasing place of scenography has brought contemporary theatre directors such as Wilson, Castellucci, Fabre to produce performances where the performance space becomes a semiotic dimension that displaces the primacy of the text. The play is not, therefore, the sole artistic vehicle for directing. This definition of directing obviously calls for a definition of what the making of the performance might be. The thesis defines the making of the performance as the activity of bringing a social event, by at least one performer, providing visual and/or textual meaning in a performance space. This definition enables us to evaluate four consistent parameters throughout theatre history: first, the social aspect associated to the performance event; second, the devising process which may be based on visual and/or textual elements; third, the presence of at least one performer in the show; fourth, the performance space (which is not simply related to the theatre stage). Although the thesis focuses primarily on theatre practice, such definition blurs the boundaries between theatre and other collaborative artistic disciplines (cinema, opera, music and dance). These parameters illustrate the possibility to undertake a generic analysis of directing, and resonate with the historical, political and artistic dimensions considered. Such a generic perspective on the role of the director addresses three significant questions: an historical question: how/why has the director emerged?; a sociopolitical question: how/why was the director a catalyst for the politicisation of theatre, and subsequently contributed to the rise of State-funded theatre policy?; and an artistic one: how/why the director has changed theatre practice and theory in the twentieth-century? Directing for the theatre as an artistic activity is a historically situated phenomenon. It would seem only natural from a contemporary perspective to associate the activity of directing to the function of the director. This is relativised, however, by the question of how the performance was produced before the modern period. The thesis demonstrates that the rise of the director is a progressive and historical phenomenon (Dort) rather than a mere invention (Viala, Sarrazac). A chronological analysis of the making of the performance throughout theatre history is the most useful way to open the study. In order to understand the emergence of the director, the research methodology assesses the interconnection of the four parameters above throughout four main periods of theatre history: the beginning of the Renaissance (meneur de jeu), the classical age (actor-manager and stage designer-manager), the modern period (director) and the contemporary period (director-facilitator, performer). This allows us properly to appraise the progressive emergence of the director, as well as to make an analysis of her/his modern and contemporary role. The first chapter argues that the physical separation between the performance space and its audience, which appeared in the early fifteenth-century, has been a crucial feature in the scenographic, aesthetic, political and social organisation of the performance. At the end of the Middle Ages, French farces which raised socio-political issues (see Bakhtin) made a clear division on a single outdoor stage (treteau) between the actors and the spectators, while religious plays (drame fiturgique, mystere) were mostly performed on various outdoor and opened multispaces. As long as the performance was liturgical or religious, and therefore confined within an acceptable framework, it was allowed. At the time, the French ecclesiastical and civil authorities tried, on several occasions, to prohibit staged performances. As a result, practitioners developed non-official indoor spaces, the Theatre de fa Trinite (1398) being the first French indoor theatre recognized by scholars. This self-exclusion from the open public space involved breaking the accepted rules by practitioners (e.g. Les Confreres de fa Passion), in terms of themes but also through individual input into a secular performance rather than the repetition of commonly known religious canvases. These developments heralded the authorised theatres that began to emerge from the mid-sixteenth century, which in some cases were subsidised in their construction. The construction of authorised indoor theatres associated with the development of printing led to a considerable increase in the production of dramatic texts for the stage. Profoundly affecting the reception of the dramatic text by the audience, the distance between the stage and the auditorium accompanied the changing relationship between practitioners and spectators. This distance gave rise to a major development of the role of the actor and of the stage designer. The second chapter looks at the significance of both the actor and set designer in the devising process of the performance from the sixteenth-century to the end of the nineteenth-century. The actor underwent an important shift in function in this period from the delivery of an unwritten text that is learned in the medieval oral tradition to a structured improvisation produced by the commedia dell 'arte. In this new form of theatre, a chef de troupe or an experienced actor shaped the story, but the text existed only through the improvisation of the actors. The preparation of those performances was, moreover, centred on acting technique and the individual skills of the actor. From this point, there is clear evidence that acting began to be the subject of a number of studies in the mid-sixteenth-century, and more significantly in the seventeenth-century, in Italy and France. This is revealed through the implementation of a system of notes written by the playwright to the actors (stage directions) in a range of plays (Gerard de Vivier, Comedie de la Fidelite Nuptiale, 1577). The thesis also focuses on Leoni de' Sommi (Quatro dialoghi, 1556 or 1565) who wrote about actors' techniques and introduced the meneur de jeu in Italy. The actor-manager (meneur de jeu), a professional actor, who scholars have compared to the director (see Strihan), trained the actors. Nothing, however, indicates that the actor-manager was directing the visual representation of the text in the performance space. From the end of the sixteenth-century, the dramatic text began to dominate the process of the performance and led to an expansion of acting techniques, such as the declamation. Stage designers carne from outside the theatre tradition and played a decisive role in the staging of religious celebrations (e.g. Actes des Apotres, 1536). In the sixteenth-century, both the proscenium arch and the borders, incorporated in the architecture of the new indoor theatres (theatre a l'italienne), contributed to create all kinds of illusions on the stage, principally the revival of perspective. This chapter shows ongoing audience demands for more elaborate visual effects on the stage. This led, throughout the classical age, and even more so during the eighteenth-century, to grant the stage design practitioner a major role in the making of the performance (see Ciceri). The second chapter demonstrates that the guidance of the actors and the scenographic conception, which are the artistic components of the role of the director, appear to have developed independently from one another until the nineteenth-century. The third chapter investigates the emergence of the director per se. The causes for this have been considered by a number of scholars, who have mainly identified two: the influence of Naturalism (illustrated by the Meiningen Company, Antoine, and Stanislavski) and the invention of electric lighting. The influence of the Naturalist movement on the emergence of the modem director in the late nineteenth-century is often considered as a radical factor in the history of theatre practice. Naturalism undoubtedly contributed to changes in staging, costume and lighting design, and to a more rigorous commitment to the harmonisation and visualisation of the overall production of the play. Although the art of theatre was dependent on the dramatic text, scholars (Osborne) demonstrate that the Naturalist directors did not strictly follow the playwright's indications written in the play in the late nineteenth-century. On the other hand, the main characteristic of directing in Naturalism at that time depended on a comprehensive understanding of the scenography, which had to respond to the requirements of verisimilitude. Electric lighting contributed to this by allowing for the construction of a visual narrative on stage. However, it was a master technician, rather than an emergent director, who was responsible for key operational decisions over how to use this emerging technology in venues such as the new Bayreuth theatre in 1876. Electric lighting reflects a normal technological evolution and cannot be considered as one of the main causes of the emergence of the director. Two further causes of the emergence of the director, not considered in previous studies, are the invention of cinema and the Symbolist movement (Lugne-Poe, Meyerhold). Cinema had an important technological influence on the practitioners of the Naturalist movement. In order to achieve a photographic truth on the stage (tableau, image), Naturalist directors strove to decorate the stage with the detailed elements that would be expected to be found if the situation were happening in reality. Film production had an influence on the work of actors (Walter). The filmmaker took over a primary role in the making of the film, as the source of the script, the filming process and the editing of the film. This role influenced the conception that theatre directors had of their own work. It is this concept of the director which influenced the development of the theatre director. As for the Symbolist movement, the director's approach was to dematerialise the text of the playwright, trying to expose the spirit, movement, colour and rhythm of the text. Therefore, the Symbolists disengaged themselves from the material aspect of the production, and contributed to give greater artistic autonomy to the role of the director. Although the emergence of the director finds its roots amongst the Naturalist practitioners (through a rigorous attempt to provide a strict visual interpretation of the text on stage), the Symbolist director heralded the modem perspective of the making of performance. The emergence of the director significantly changed theatre practice and theory. For instance, the rehearsal period became a clear work in progress, a platform for both developing practitioners' techniques and staging the show. This chapter explores and contrasts several practitioners' methods based on the two aspects proposed for the definition of the director (guidance of the actors and materialisation of a visual space). The fourth chapter argues that the role of the director became stronger, more prominent, and more hierarchical, through a more political and didactic approach to theatre as exemplified by the cases of France and Germany at the end of the nineteenth-century and through the First World War. This didactic perspective to theatre defines the notion of political theatre. Political theatre is often approached by the literature (Esslin, Willett) through a Marxist interpretation of the great German directors' productions (Reinhardt, Piscator, Brecht). These directors certainly had a great influence on many directors after the Second World War, such as Jean Vilar, Judith Molina, Jean-Louis Barrault, Roger Planchon, Augusto Boal, and others. This chapter demonstrates, moreover, that the director was confirmed through both ontological and educational approaches to the process of making the performance, and consequently became a central and paternal figure in the organisational and structural processes practiced within her/his theatre company. In this way, the stance taken by the director influenced the State authorities in establishing theatrical policy. This is an entirely novel scholarly contribution to the study of the director. The German and French States were not indifferent to the development of political theatre. A network of public theatres was thus developed in the inter-war period, and more significantly after the Second World War. The fifth chapter shows how State theatre policies establish its sources in the development of political theatre, and more specifically in the German theatre trade union movement (Volksbiihne) and the great directors at the end of the nineteenth-century. French political theatre was more influenced by playwrights and actors (Romain Rolland, Louise Michel, Louis Lumet, Emile Berny). French theatre policy was based primarily on theatre directors who decentralised their activities in France during both the inter-war period and the German occupation. After the Second World War, the government established, through directors, a strong network of public theatres. Directors became both the artistic director and the executive director of those institutionalised theatres. The institution was, however, seriously shaken by the social and political upheaval of 1968. It is the link between the State and the institution in which established directors were entangled that was challenged by the young emerging directors who rejected institutionalised responsibility in favour of the autonomy of the artist in the 1960s. This process is elucidated in chapter five. The final chapter defines the contemporary role of the director in contrasting thework of a number of significant young theatre practitioners in the 1960s such as Peter Brook, Ariane Mnouchkine, The Living Theater, Jerzy Grotowski, Augusto Boal, Eugenio Barba, all of whom decided early on to detach their companies from any form of public funding. This chapter also demonstrates how they promoted new forms of performance such as the performance of the self. First, these practitioners explored new performance spaces outside the traditional theatre building. Producing performances in a non-dedicated theatre place (warehouse, street, etc.) was a more frequent practice in the 1960s than before. However, the recent development of cybertheatre questions both the separation of the audience and the practitioners and the place of the director's role since the 1990s. Secondly, the role of the director has been multifaceted since the 1960s. On the one hand, those directors, despite all their different working methods, explored western and non-western acting techniques based on both personal input and collective creation. They challenged theatrical conventions of both the character and the process of making the performance. On the other hand, recent observations and studies distinguish the two main functions of the director, the acting coach and the scenographe, both having found new developments in cinema, television, and in various others events. Thirdly, the contemporary director challenges the performance of the text. In this sense, Antonin Artaud was a visionary. His theatre illustrates the need for the consideration of the totality of the text, as well as that of theatrical production. By contrasting the theories of Artaud, based on a non-dramatic form of theatre, with one of his plays (Le Jet de Sang), this chapter demonstrates how Artaud examined the process of making the performance as a performance. Live art and autobiographical performance, both taken as directing the se(f, reinforce this suggestion. Finally, since the 1990s, autobiographical performance or the performance of the self is a growing practical and theoretical perspective in both performance studies and psychology-related studies. This relates to the premise that each individual is making a representation (through memory, interpretation, etc.) of her/his own life (performativity). This last section explores the links between the place of the director in contemporary theatre and performers in autobiographical practices. The role of the traditional actor is challenged through non-identification of the character in the play, while performers (such as Chris Burden, Ron Athey, Orlan, Franko B, Sterlac) have, likewise, explored their own story/life as a performance. The thesis demonstrates the validity of the four parameters (performer, performance space, devising process, social event) defining a generic approach to the director. A generic perspective on the role of the director would encompass: a historical dimension relative to the reasons for and stages of the 'emergence' of the director; a socio-political analysis concerning the relationship between the director, her/his institutionalisation, and the political realm; and the relationship between performance theory, practice and the contemporary role of the director. Such a generic approach is a new departure in theatre research and might resonate in the study of other collaborative artistic practices.
Resumo:
Self-similar optical pulses (or “similaritons”) of parabolic intensity profile can be found as asymptotic solutions of the nonlinear Schr¨odinger equation in a gain medium such as a fiber amplifier or laser resonator. These solutions represent a wide-ranging significance example of dissipative nonlinear structures in optics. Here, we address some issues related to the formation and evolution of parabolic pulses in a fiber gain medium by means of semi-analytic approaches. In particular, the effect of the third-order dispersion on the structure of the asymptotic solution is examined. Our analysis is based on the resolution of ordinary differential equations, which enable us to describe the main properties of the pulse propagation and structural characteristics observable through direct numerical simulations of the basic partial differential equation model with sufficient accuracy.
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This paper employs a Component GARCH in Mean model to show that house prices across a number of major US cities between 1987 and 2009 have displayed asset market properties in terms of both risk-return relationships and asymmetric adjustment to shocks. In addition, tests for structural breaks in the mean and variance indicate structural instability across the data range. Multiple breaks are identified across all cities, particularly for the early 1990s and during the post-2007 financial crisis as housing has become an increasingly risky asset. Estimating the models over the individual sub-samples suggests that over the last 20 years the financial sector has increasingly failed to account for the levels of risk associated with real estate markets. This result has possible implications for the way in which financial institutions should be regulated in the future.
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Previous empirical assessments of the effectiveness of structural merger remedies have focused mainly on the subsequent viability of the divested assets. Here, we take a different approach by examining how competitive are the market structures which result from the divestments. We employ a tightly specified sample of markets in which the European Commission (EC) has imposed structural merger remedies. It has two key features: (i) it includes all mergers in which the EC appears to have seriously considered, simultaneously, the possibility of collective dominance, as well as single dominance; (ii) in a previous paper, for the same sample, we estimated a model which proved very successful in predicting the Commission’s merger decisions, in terms of the market shares of the leading firms. The former allows us to explore the choices between alternative theories of harm, and the latter provides a yardstick for evaluating whether markets are competitive or not – at least in the eyes of the Commission. Running the hypothetical post-remedy market shares through the model, we can predict whether the EC would have judged the markets concerned to be competitive, had they been the result of a merger rather than a remedy. We find that a significant proportion were not competitive in this sense. One explanation is that the EC has simply been inconsistent – using different criteria for assessing remedies from those for assessing the mergers in the first place. However, a more sympathetic – and in our opinion, more likely – explanation is that the Commission is severely constrained by the pre-merger market structures in many markets. We show that, typically, divestment remedies return the market to the same structure as existed before the proposed merger. Indeed, one can argue that any competition authority should never do more than this. Crucially, however, we find that this pre-merger structure is often itself not competitive. We also observe an analogous picture in a number of markets where the Commission chose not to intervene: while the post-merger structure was not competitive, nor was the pre-merger structure. In those cases, however, the Commission preferred the former to the latter. In effect, in both scenarios, the EC was faced with a no-win decision. This immediately raises a follow-up question: why did the EC intervene for some, but not for others – given that in all these cases, some sort of anticompetitive structure would prevail? We show that, in this sample at least, the answer is often tied to the prospective rank of the merged firm post-merger. In particular, in those markets where the merged firm would not be the largest post-merger, we find a reluctance to intervene even where the resulting market structure is likely to be conducive to collective dominance. We explain this by a willingness to tolerate an outcome which may be conducive to tacit collusion if the alternative is the possibility of an enhanced position of single dominance by the market leader. Finally, because the sample is confined to cases brought under the ‘old’ EC Merger Regulation, we go on to consider how, if at all, these conclusions require qualification following the 2004 revisions, which, amongst other things, made interventions for non-coordinated behaviour possible without requiring that the merged firm be a dominant market leader. Our main conclusions here are that the Commission appears to have been less inclined to intervene in general, but particularly for Collective Dominance (or ‘coordinated effects’ as it is now known in Europe as well as the US.) Moreover, perhaps contrary to expectation, where the merged firm is #2, the Commission has to date rarely made a unilateral effects decision and never made a coordinated effects decision.
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It is generally believed that the structural reforms that were introduced in India following the macro-economic crisis of 1991 ushered in competition and forced companies to become more efficient. However, whether the post-1991 growth is an outcome of more efficient use of resources or greater use of factor inputs remains an open empirical question. In this paper, we use plant-level data from 1989–1990 and 2000–2001 to address this question. Our results indicate that while there was an increase in the productivity of factor inputs during the 1990s, most of the growth in value added is explained by growth in the use of factor inputs. We also find that median technical efficiency declined in all but one of the industries between 1989–1990 and 2000–2001, and that change in technical efficiency explains a very small proportion of the change in gross value added.
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In this paper we present a simple three-sector model explaining the structural change in employment, which is a modified version of Rowthorn-Wells (1987). We supplement the theoretical analysis with simple econometric tests, which illustrate how the modified Rowthorn-Wells model can be used to (i) motivate empirical estimates of the link between the level of development and structures of employment, (ii) illustrate structural distortions under the command economies, and the structural adjustment that happened during the post-Communist transition. We demonstrate that in the case of these economies, the transition process leads to an adjustment to the employment structures predicted by the model.
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Using a data set for the 162 largest Hungarian firms during the period of 1994-1999, this paper explores the determinants of equity shares held by both foreign investors and Hungarian corporations. Evidence is found for a post-privatisation evolution towards more homogeneous equity structures, where dominant categories of Hungarian and foreign owners aim at achieving controlling stakes. In addition, focusing on firm-level characteristics we find that exporting firms attract foreign owners who acquire controlling equity stakes. Similarly, firm-size measurements are positively associated with the presence of foreign investors. However, they are negatively associated with 100% foreign ownership, possibly because the marginal costs of acquiring additional equity are growing with the size of the assets. The results are interpreted within the framework of the existing theory. In particular, following Demsetz and Lehn (1985) and Demsetz and Villalonga (2001) we argue that equity should not be treated as an exogenous variable. As for specific determinants of equity levels, we focus on informational asymmetries and (unobserved) ownership-specific characteristics of foreign investors and Hungarian investors.
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An organisation’s ability to internalise external knowledge and learn from various sources in undertaking new product development and/or entering a new market is crucial to its competitive performance. Nevertheless, little attention has been paid to how growth-oriented small firms identify and exploit entrepreneurial opportunities, i.e. take entrepreneurial action, related to such methods of development, in a mature, contracting industry. The latter is particularly relevant to recent discussion and debate in academic and policy-making circles concerning the salvage of the clothing manufacture industry in developed industrialised countries, by intensifying efforts relating to diversification into high value manufacturing sectors. This paper, based on an instrumental case-firm, demonstrates analytically how learning as entrepreneurial action relating to diversifying into /technical clothing – i.e. a high value manufacturing/innovatory sector - takes place, drawing on situated learning theory. It is argued that learning relating to such entrepreneurial action is dynamic in nature and is founded on specific organising principles that foster both inter- and intracommunal learning.
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This paper proposes a more profound discussion of the philosophical underpins of sustainability than currently exists in the MOT literature and considers their influence on the construction of the theories on green operations and technology management. Ultimately, it also debates the link between theory and practice on this subject area. The paper is derived from insights gained in three research projects completed during the past twelve years, primarily involving the first author. From 2000 to 2002, an investigation using scenario analysis, aimed at reducing atmospheric pollution in urban centres by substituting natural gas for petrol and diesel, provided the first set of insights about public policy, environmental impacts, investment analysis, and technological feasibility. The second research project, from 2003 to 2005, using a survey questionnaire, was aimed at improving environmental performance in livestock farming and explored the issues of green supply chain scope, environmental strategy and priorities. Finally, the third project, from 2006 to 2011, investigated environmental decisions in manufacturing organisations through case study research and examined the underlying sustainability drivers and decision-making processes. By integrating the findings and conclusions from these projects, the link between philosophy, theory, and practice of green operations and technology management is debated. The findings from all these studies show that the philosophical debate seems to have little influence on theory building so far. For instance, although ‘sustainable development’ emphasises ‘meeting the needs of current and future generation’, no theory links essentiality and environmental impacts. Likewise, there is a weak link between theory and the practical issues of green operations and technology management. For example, the well-known ‘life-cycle analysis’ has little application in many cases because the life cycle of products these days is dispersed within global production and consumption systems and there are different stakeholders for each life cycle stage. The results from this paper are relevant to public policy making and corporate environmental strategy and decision making. Most of the past and current studies in the subject of green operations and sustainability management deal with only a single sustainability dimension at any one time. Here the value and originality of this paper lies in its integration between philosophy, theory, and practice of green technology and operations management.
Resumo:
Self-similar optical pulses (or “similaritons”) of parabolic intensity profile can be found as asymptotic solutions of the nonlinear Schr¨odinger equation in a gain medium such as a fiber amplifier or laser resonator. These solutions represent a wide-ranging significance example of dissipative nonlinear structures in optics. Here, we address some issues related to the formation and evolution of parabolic pulses in a fiber gain medium by means of semi-analytic approaches. In particular, the effect of the third-order dispersion on the structure of the asymptotic solution is examined. Our analysis is based on the resolution of ordinary differential equations, which enable us to describe the main properties of the pulse propagation and structural characteristics observable through direct numerical simulations of the basic partial differential equation model with sufficient accuracy.
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The primary aim of this thesis was to investigate the in vivo ocular morphological and contractile changes occurring within the accommodative apparatus prior to the onset of presbyopia, with particular reference to ciliary muscle changes with age and the origin of a myopic shift in refraction during incipient presbyopia. Commissioned semi-automated software proved capable of extracting accurate and repeatable measurements from crystalline lens and ciliary muscle Anterior Segment Optical Coherence Tomography (AS-OCT) images and reduced the subjectivity of AS-OCT image analysis. AS-OCT was utilised to document longitudinal changes in ciliary muscle morphology within an incipient presbyopic population (n=51). A significant antero-inwards shift of ciliary muscle mass was observed after 2.5 years. Furthermore, in a subgroup study (n=20), an accommodative antero-inwards movement of ciliary muscle mass was evident. After 2.5 years, the centripetal response of the ciliary muscle significantly attenuated during accommodation, whereas the antero-posterior mobility of the ciliary muscle remained invariant. Additionally, longitudinal measurement of ocular biometry revealed a significant increase in crystalline lens thickness and a corresponding decrease in anterior chamber depth after 2.5 years (n=51). Lenticular changes appear to be determinant of changes in refraction during incipient presbyopia. During accommodation, a significant increase in crystalline lens thickness and axial length was observed, whereas anterior chamber depth decreased (n=20). The change in ocular biometry per dioptre of accommodation exerted remained invariant after 2.5 years. Cross-sectional ocular biometric data were collected to quantify accommodative axial length changes from early adulthood to advanced presbyopia (n=72). Accommodative axial length elongation significantly attenuated during presbyopia, which was consistent with a significant increase in ocular rigidity during presbyopia. The studies presented in this thesis support the Helmholtz theory of accommodation and despite the reduction in centripetal ciliary muscle contractile response with age, primarily implicate lenticular changes in the development of presbyopia.
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The basic conceptions of the model „entity-relationship” as entities, relationships, structural constraints of the relationships (index cardinality, participation degree, and structural constraints of kind (min, max)) are considered and formalized in terms of relations theory. For the binary relations two operators (min and max) are introduced; structural constraints are determined in terms of the operators; the main theorem about compatibility of these operators’ values on the source relation and inversion to it is given here.
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Full text: The idea of producing proteins from recombinant DNA hatched almost half a century ago. In his PhD thesis, Peter Lobban foresaw the prospect of inserting foreign DNA (from any source, including mammalian cells) into the genome of a λ phage in order to detect and recover protein products from Escherichia coli [ 1 and 2]. Only a few years later, in 1977, Herbert Boyer and his colleagues succeeded in the first ever expression of a peptide-coding gene in E. coli — they produced recombinant somatostatin [ 3] followed shortly after by human insulin. The field has advanced enormously since those early days and today recombinant proteins have become indispensable in advancing research and development in all fields of the life sciences. Structural biology, in particular, has benefitted tremendously from recombinant protein biotechnology, and an overwhelming proportion of the entries in the Protein Data Bank (PDB) are based on heterologously expressed proteins. Nonetheless, synthesizing, purifying and stabilizing recombinant proteins can still be thoroughly challenging. For example, the soluble proteome is organized to a large part into multicomponent complexes (in humans often comprising ten or more subunits), posing critical challenges for recombinant production. A third of all proteins in cells are located in the membrane, and pose special challenges that require a more bespoke approach. Recent advances may now mean that even these most recalcitrant of proteins could become tenable structural biology targets on a more routine basis. In this special issue, we examine progress in key areas that suggests this is indeed the case. Our first contribution examines the importance of understanding quality control in the host cell during recombinant protein production, and pays particular attention to the synthesis of recombinant membrane proteins. A major challenge faced by any host cell factory is the balance it must strike between its own requirements for growth and the fact that its cellular machinery has essentially been hijacked by an expression construct. In this context, Bill and von der Haar examine emerging insights into the role of the dependent pathways of translation and protein folding in defining high-yielding recombinant membrane protein production experiments for the common prokaryotic and eukaryotic expression hosts. Rather than acting as isolated entities, many membrane proteins form complexes to carry out their functions. To understand their biological mechanisms, it is essential to study the molecular structure of the intact membrane protein assemblies. Recombinant production of membrane protein complexes is still a formidable, at times insurmountable, challenge. In these cases, extraction from natural sources is the only option to prepare samples for structural and functional studies. Zorman and co-workers, in our second contribution, provide an overview of recent advances in the production of multi-subunit membrane protein complexes and highlight recent achievements in membrane protein structural research brought about by state-of-the-art near-atomic resolution cryo-electron microscopy techniques. E. coli has been the dominant host cell for recombinant protein production. Nonetheless, eukaryotic expression systems, including yeasts, insect cells and mammalian cells, are increasingly gaining prominence in the field. The yeast species Pichia pastoris, is a well-established recombinant expression system for a number of applications, including the production of a range of different membrane proteins. Byrne reviews high-resolution structures that have been determined using this methylotroph as an expression host. Although it is not yet clear why P. pastoris is suited to producing such a wide range of membrane proteins, its ease of use and the availability of diverse tools that can be readily implemented in standard bioscience laboratories mean that it is likely to become an increasingly popular option in structural biology pipelines. The contribution by Columbus concludes the membrane protein section of this volume. In her overview of post-expression strategies, Columbus surveys the four most common biochemical approaches for the structural investigation of membrane proteins. Limited proteolysis has successfully aided structure determination of membrane proteins in many cases. Deglycosylation of membrane proteins following production and purification analysis has also facilitated membrane protein structure analysis. Moreover, chemical modifications, such as lysine methylation and cysteine alkylation, have proven their worth to facilitate crystallization of membrane proteins, as well as NMR investigations of membrane protein conformational sampling. Together these approaches have greatly facilitated the structure determination of more than 40 membrane proteins to date. It may be an advantage to produce a target protein in mammalian cells, especially if authentic post-translational modifications such as glycosylation are required for proper activity. Chinese Hamster Ovary (CHO) cells and Human Embryonic Kidney (HEK) 293 cell lines have emerged as excellent hosts for heterologous production. The generation of stable cell-lines is often an aspiration for synthesizing proteins expressed in mammalian cells, in particular if high volumetric yields are to be achieved. In his report, Buessow surveys recent structures of proteins produced using stable mammalian cells and summarizes both well-established and novel approaches to facilitate stable cell-line generation for structural biology applications. The ambition of many biologists is to observe a protein's structure in the native environment of the cell itself. Until recently, this seemed to be more of a dream than a reality. Advances in nuclear magnetic resonance (NMR) spectroscopy techniques, however, have now made possible the observation of mechanistic events at the molecular level of protein structure. Smith and colleagues, in an exciting contribution, review emerging ‘in-cell NMR’ techniques that demonstrate the potential to monitor biological activities by NMR in real time in native physiological environments. A current drawback of NMR as a structure determination tool derives from size limitations of the molecule under investigation and the structures of large proteins and their complexes are therefore typically intractable by NMR. A solution to this challenge is the use of selective isotope labeling of the target protein, which results in a marked reduction of the complexity of NMR spectra and allows dynamic processes even in very large proteins and even ribosomes to be investigated. Kerfah and co-workers introduce methyl-specific isotopic labeling as a molecular tool-box, and review its applications to the solution NMR analysis of large proteins. Tyagi and Lemke next examine single-molecule FRET and crosslinking following the co-translational incorporation of non-canonical amino acids (ncAAs); the goal here is to move beyond static snap-shots of proteins and their complexes and to observe them as dynamic entities. The encoding of ncAAs through codon-suppression technology allows biomolecules to be investigated with diverse structural biology methods. In their article, Tyagi and Lemke discuss these approaches and speculate on the design of improved host organisms for ‘integrative structural biology research’. Our volume concludes with two contributions that resolve particular bottlenecks in the protein structure determination pipeline. The contribution by Crepin and co-workers introduces the concept of polyproteins in contemporary structural biology. Polyproteins are widespread in nature. They represent long polypeptide chains in which individual smaller proteins with different biological function are covalently linked together. Highly specific proteases then tailor the polyprotein into its constituent proteins. Many viruses use polyproteins as a means of organizing their proteome. The concept of polyproteins has now been exploited successfully to produce hitherto inaccessible recombinant protein complexes. For instance, by means of a self-processing synthetic polyprotein, the influenza polymerase, a high-value drug target that had remained elusive for decades, has been produced, and its high-resolution structure determined. In the contribution by Desmyter and co-workers, a further, often imposing, bottleneck in high-resolution protein structure determination is addressed: The requirement to form stable three-dimensional crystal lattices that diffract incident X-ray radiation to high resolution. Nanobodies have proven to be uniquely useful as crystallization chaperones, to coax challenging targets into suitable crystal lattices. Desmyter and co-workers review the generation of nanobodies by immunization, and highlight the application of this powerful technology to the crystallography of important protein specimens including G protein-coupled receptors (GPCRs). Recombinant protein production has come a long way since Peter Lobban's hypothesis in the late 1960s, with recombinant proteins now a dominant force in structural biology. The contributions in this volume showcase an impressive array of inventive approaches that are being developed and implemented, ever increasing the scope of recombinant technology to facilitate the determination of elusive protein structures. Powerful new methods from synthetic biology are further accelerating progress. Structure determination is now reaching into the living cell with the ultimate goal of observing functional molecular architectures in action in their native physiological environment. We anticipate that even the most challenging protein assemblies will be tackled by recombinant technology in the near future.