This work is the result of evaluating a novel strategy for the purification of recombinant His-tagged proteins. Proteins purified in this study were the E. coli translational proteins IF-3, RF-1, and RFF. The study aimed to analyse the potential of using Bio-Works WorkBeads™40 TREN, a multimodal anion ion exchange chromatography resin, as a pretreatment step upstream an immobilized metal ion chromatography (IMAC) resin to enhance performance efficiency of His-tagged protein purification. The method demonstrated here shows potential for anyone seeking to increase the purity of His-tagged protein purification or to introduce an effective purification procedure by replacing a polishing step downstream IMAC with WorkBeads 40 TREN upstream IMAC. The latter contributing to guard the IMAC column from heavy bioburden. This study showed that running WorkBeads 40 TREN prior IMAC captures impurities and removes 97-98 % more dsDNA compared to direct IMAC. WorkBeads 40 TREN is therefore highly advantageous to include early in a purification process to remove protein binding DNA fragments. Moreover, WorkBeads 40 TREN increases purity in the final product by capturing more host cell proteins than when running direct IMAC. This concept is general and WorkBeads 40 TREN could be used upstream a variety of resins such as Protein A and RPC.

This work aims to contribute to the research of Czech elites of the 19th century and to write up the hitherto neglected topic of professional and social rise of Czech architects and builders in the 19th century. At this time, as a result of state reforms of the educational, administrative and legal system, the professional perception and social status of this class changed. In the introduction, architects and builders are included in the process of modernization of the construction industry and defined on the basis of their relationship to art, engineering and business. Subsequently, architects and builders are defined as a socio-professional group that has reached a certain degree of professionalism during this period. Against the background of the formation of a modern system of building and architectural education, law and administration, individual features of professionalization are monitored on specific cases of architects and builders using the method of collective biography of elites in research areas of their social background, education, construction practice and other public activities. Thus, two levels intersect in the work - general and personal. In general, the interconnection of construction education, law and administration since the establishment of construction disciplines at...

Ce mémoire, qui s’intéresse aux enjeux de l’imbrication du philosophique et du romanesque chez les penseurs des Lumières françaises, cherchera à mettre de l’avant la contribution originale de Denis Diderot à la réflexion philosophique sur le roman, de même qu’à l’exploration des ressources littéraires permettant de philosopher par le roman. Nous nous proposons, dans un premier temps, d’examiner ce qui relève d'une théorisation explicite de l'écriture des fictions narratives dans l’oeuvre du philosophe langrois. Nous nous attacherons à montrer comment l’approfondissement des thèses de son matérialisme, ainsi que les déceptions consécutives à ses projets de réforme du théâtre, sont venus bouleverser les conceptions esthétiques et morales qui formaient le premier horizon normatif à partir duquel Diderot envisageait les modalités idéales de l'écriture et de la réception romanesques. Puis, dans un deuxième temps, nous chercherons à faire voir comment Diderot, à titre d’auteur de fictions narratives, entreprend une exploration aussi originale que féconde des ressorts proprement philosophiques de l’écriture romanesque. Les dernières sections de notre étude seront consacrées à une lecture approfondie de Jacques le fataliste et son maître. Nous tâcherons d’abord d’en dégager les principales ressources mises au service de la formation d’un lecteur de romans plus lucide et plus critique. Puis, nous chercherons à mettre en lumière de quelle façon cette propédeutique, si elle réussit, doit ultimement conduire le lecteur à entamer un véritable dialogue avec l’oeuvre, tant sur les questions métaphysiques et ontologiques qu’entraîne le « fatalisme » de Jacques, que sur les enjeux moraux qui en découlent. / This thesis, which deals with the interweaving of philosophy and literature in the work of French Enlightenment thinkers, seeks to shed light on Denis Diderot’s unique contributions to philosophical reflection on the novel, as well as to the exploration of the literary resources that make it possible to delve into philosophical issues through the novel. Beginning with an examination of Diderot’s explicit theories about writing narrative fiction, it argues that the outgrowth of his materialism and the disenchantment following his attempts at theatre reform upended the aesthetic and moral basis for the initial normative framework underpinning his analysis of how fiction is ideally written and received. It goes on to examine how Diderot, as a novelist, undertook an innovative and fruitful exploration of the unsuspected philosophical resources that narrative fiction offers. The final sections are devoted to a close reading of Jacques the Fatalist and His Master, beginning with an attempt to identify the main resources it deploys to train more lucid and critical readers of fiction. The thesis then seeks to show how this knowledge base, if successfully transmitted, ultimately leads the reader to engage in a genuine dialogue with the work, both in terms of the metaphysical and ontological issues raised by Jacques’ “fatalism” and the moral issues that ensue.

Im ersten Halbjahr 2016 wurde in Kooperation mit dem Martinsclub Bremen e.V. eine empirische Untersuchung zur funktionalen Varietät ' Leichte Sprache ' durchgeführt. Diese Ziele wurden verfolgt: 1) Für die ' Leichte Sprache ' wird u.a. vom ' Netzwerk Leichte Sprache ' reklamiert, für eine äußerst heterogene Zielgruppe von Nutzen zu sein. In der Untersuchung wird an Testitems, die z.T. erheblich gegen die ' Regeln ' für ' Leichte Sprache ' verstoßen, ermittelt werden, wie Gewährspersonen unterschiedlicher Zielgruppen (Menschen mit kognitiver Beeinträchtigung, Funktionale Analphabeten und Migranten) die ‚Schwierigkeit' und die ‚Akzeptabilität' der Items bewerten. 2) Die Regeln für ' Leichte Sprache ' sind (linguistisch) zu explizieren und auf die Bedürfnisse unterschiedlicher Zielgruppen zuzuschneiden. Beide Ziele wurden durch die Untersuchung für einzelne morphosyntaktische Phänomene erreicht. In diesem Artikel werden die Ergebnisse der Studie knapp vorgestellt und am Beispiel passivischer Strukturen des Deutschen im Detail diskutiert. Angrenzende Phänomenbereiche (sein-Perfekt, Kopulakonstruktionen mit sein und werden sowie werden mit futurischer und/oder epistemischer Lesart) werden nur angeschnitten.

The period 1889-1891 has been regarded as crucial in Oscar Wilde’s (1854-1900) career. Having been somewhat unsuccessful as a writer during the 1880s, and turning to journalism to earn a living, Wilde in this period saw the publication of his dialogues which led to his sole novel The Picture of Dorian Gray (hereafter, Dorian), serialised in 1890 before being republished as a novel in 1891. It has been characterized as a turning point in his career, and critics have studied these works in detail, as well as those which followed, especially the four society’s comedies: Lady Windermere’s Fan (1892), A Woman of No Importance (1893), An Ideal Husband (1895), The Importance of Being Earnest (1895). However, besides this selection, much of Wilde’s work remains under-researched, particularly his fairy tales, whose study suffers from the prejudicial categorisation as children’s literature. Research to date has tended to privilege a single aspect of Wilde’sfairy tales, as in Jarlath Killeen’s The Fairy Tales of Oscar Wilde (2007) focusing on a Victorian societal perspective, rather than studying the fairy tales as a coherent and integral part of Wilde’s lifework. The essay focuses on ‘The Fisherman and His Soul’ (hereafter, ‘Fisherman’), recognised as the most sophisticated tale from Wilde’s A House of Pomegranates (1891) (hereafter, Pomegranates). It will establish the essential role of ‘Fisherman’ in understanding Wilde’s complex aesthetic philosophy by examining the tale from two distinct levels of intertextuality. First, Wilde borrowed some of the most emblematic aesthetical and narrative elements from Andersen’s ‘The Little Mermaid’ (hereafter, ‘Mermaid’) and other similar fairy tales from the early nineteenth century for explicitly positioning ‘Fisherman’ as a response to Andersen’stale. Taking the opposite approach to Andersen’s ‘Mermaid’, Wilde’s ‘Fisherman’ stands as a social critique against the Victorian doxa, especially denouncing its nefarious effect on art. Second, through epistolary and textual evidence, the essay reveals the connections between ‘Fisherman’, Dorian, and ‘The Soul of Man’ (1891), including their genesis, design, themes, and discourse. This dual intertextuality of ‘Fisherman’ allows us to reassess Wilde’s tale as an influential text. It contributes simultaneously to comprehending better how consistent Wilde’s aesthetic standards and societal view were.

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Systematic studies of basal nonsense suppression, orthogonality of tRNAPyl variants, and cross recognition between codons and tRNA anticodons are reported. E. coli displays detectable basal amber and opal suppression but shows a negligible ochre suppression. Although detectable, basal amber suppression is fully inhibited when a pyrrolysyl-tRNA synthetase (PylRS)-tRNAPyl_CUA pair is genetically encoded. trnaPyl_CUA is aminoacylated by an E. coli aminoacyl-tRNA synthetase at a low level, however, this misaminoacylation is fully inhibited when both PylRS and its substrate are present. Besides that it is fully orthogonal in E. coli and can be coupled with PylRS to genetically incorporate a NAA at an ochre codon, tRNAPyl_UUA is not able to recognize an UAG codon to induce amber suppression. This observation is in direct conflict with the wobble base pair hypothesis and enables using an evolved M. jannaschii tyrosyl-tRNA synthetase-tRNAPyl_UUA pair and the wild type or evolved PylRS-tRNAPyl_UUA pair to genetically incorporate two different NAAs at amber and ochre codons. tRNAPyl_UCA is charged by E. coli tryptophanyl-tRNA synthetase, thus not orthogonal in E. coli. Mutagenic studies of trnaPyl_UCA led to the discovery of its G73U form which shows a higher orthogonality. Mutating trnaPyl_CUA to trnaPyl_UCCU not only leads to the loss of the relative orthogonality of tRNAPyl in E. coli but also abolishes its aminoacylation by PylRS.

Spannmål med varierande fuktighetsgrad kan lagras i kvadratiska silor med väggelement bestående av korrugerad plåt. Då det lagrade spannmålet ska tömmas har det fuktiga spannmålet en tendens att fastna på väggarna, vilket medför att spannmålet i efterhand måste avlägsnas manuellt. För att slippa denna tidskrävande process monteras en slät plåt på den korrugerade plåten, för att på så vis hindra det fuktiga spannmålet från att fastna. Om samma plåtdimensioner för den korrugerade plåten används då den släta plåten är och inte är monterad kan väggelementen utsättas för buckling. Det här examensarbetet handlar således om hur väggelementen ska dimensioneras för att strukturen inte ska utsättas för buckling. De silor som undersöks har tvärsnitten 3,0 x 3,0 m och 2,5 x 2,5 m med väggelement som består av endast korrugerad plåt samt med slät plåt monterad på korrugerad plåt. Vidare har silorna vertikala väggar med höjden 8,4 m som består av tio sektioner. Beräkningar har utförts då fyllnadsmaterialet är vete.   För att väggelementen ska kunna dimensioneras beräknas vilka trycksituationer som uppstår i de olika silorna med hjälp av den svenska och europeiska standarden Eurokod (2006), EN 1991-4 för tryckberäkningar i silor och behållare. För att beräkna trycksituationerna delas de aktuella silorna in i åtgärdsklass 1 då deras kapacitet understiger 100 ton, vilket innebär att osymmetriska tryck kan ignoreras. Silorna klassas även som slanka. Då silornas utlopp består av en pyramidformad tratt med centriskt utlopp och då trattens halva inre vinkel är 45° uppstår ett inre rörflöde vid tömning. Detta medför enligt Eurokod att silorna ska dimensioneras enligt trycken som uppstår vid fyllning. Det horisontella och vertikala trycket samt trycket som uppstår från friktionen beräknas för de olika tvärsnitten.   CAD-programmet Pro Engineer och finita elementtillägget Mechanica används för att modellera de aktuella silorna och utföra analyser med avseende på spänning och buckling. Modellerna har fyra symmetriplan, därför modelleras endast en åttondel av de aktuella strukturerna. Detta motsvarar ett halvt väggelement och en halv stolpe. Modellerna skapas som skalmodeller och randvillkor ansätts i alla snittytor samt på stolpens över och underkant. I modellerna förenklas strukturen genom att inga skruvar eller radier modelleras. Trycken framräknade enligt Eurokod räknas om till krafter och appliceras på modellerna. Hela strukturen modelleras i stål med sträckgränsen 180 MPa.   En av företagets äldre dimensioneringsstandarder undersöks genom att väggelementen tilldelas plåttjocklekar enligt denna. För att undersöka var de mest kritiska områdena för buckling uppstår utförs en bucklingsanalys grundad på en statisk analys för modellerna. Resultatet från bucklingsanalysen för silon med väggelement bestående av korrugerad plåt med väggbredden 3,0 m visar att buckling uppstår på den näst nedersta sektionen vid 72 % av den applicerade kraftresultanten. För silon med samma väggelement fast med bredden 2,5 m uppstår buckling på den översta sektionen där bucklingskraften uppgår till 62 % av den applicerade kraftresultanten. För silorna med väggelement bestående av slät plåt monterad på korrugerad plåt uppstår buckling redan vid 3-4 % av den applicerade kraftresultanten, för båda väggbredderna. Då det genom analyser framkommit att det är lokala bucklingar enbart på den släta plåten som ger dessa låga värden friläggs den korrugerade plåten för att trycksituationen som verkar från den släta plåten på så vis ska kunna erhållas. Med kraften från den släta väggen kan den korrugerade plåten därmed dimensioneras mot buckling. Vid bucklingsanalys av den korrugerade plåten som är tryckbelastad av den släta plåten framgår att buckling då väggbredden är 3,0 m uppstår på silons näst understa sektion. Bucklingskraften uppgår till 59 % av den applicerade kraftresultanten. För silon med väggbredden 2,5 m uppstår buckling vid 51 % av den applicerade kraftresultanten på silons översta sektion.   Då målet är att de korrugerade plåtarna inte får utsättas för buckling provas plåttjocklekar fram i finita elementmodellerna tills att bucklingskraften uppgår till minst 110 % av den applicerade kraftresultanten. Detta genererar att plåttjocklekarna för de nedre fyra sektionerna av väggelementen bestående av korrugerad plåt med bredden 3,0 m behöver ökas. För silon med samma väggelement fast med väggbredden 2,5 m behöver dimensionen på de två översta sektionerna ökas. För silorna med väggelement bestående av slät plåt monterad på korrugerad plåt behöver den korrugerade plåten för väggbredden 3,0 m ökas för de fem understa sektionerna medan samma dimensioner för väggbredden 2,5 m kan användas som för silon med väggelement bestående av enbart korrugerad plåt. Om den släta plåten inte får utsättas för buckling behöver plåttjocklekarna ökas från mellan 5,5 mm och 1,5 mm. Vid beräkningar av plåtdimensioner har ett konservativ förfarande använts vilket innebär att företagets dimensionering kan vara korrekt. Erhållna dimensioneringsförslag är emellertid rimliga för de korrugerade plåtarna. / Grains with varying humidity can be stored in square silos with wall elements consisting of corrugated sheet. When the stored grain is to be emptied from the silos it has tendency to stick to the walls, especially if humid, which means that the grain must be removed manually. To avoid this time-consuming process a flat sheet is mounted on the corrugated sheet to prevent the moist grain from sticking to the wall. If the same dimension on the corrugated sheet is used when the flat sheet is or is not mounted the walls may be subjected to buckling. This thesis is thus about how the wall elements shall be designed in order to prevent buckling. The silos that have been examined have a cross section of 3.0 x 3.0 m and 2.5 x 2.5 m respectively with wall elements consisting of only corrugated sheet or smooth sheet mounted on corrugated sheet. Furthermore, the silos got vertical walls with a height of 8.4 m consisting of ten sections. Calculations are made with wheat as the stored grain.   To be able to dimension the wall elements the pressure is calculated for the different silos, using the Swedish and European standard Eurocode (2006), EN 1991-4 for pressure calculations in silos and tanks. To calculate the pressure the silos are assigned into action assessment class 1, since their capacity are less than a 100 tons, which further means that the unsymmetrical pressure can be ignored. The silos are also classified as slender. As the silos outlet consists of a square pyramidal hopper with centric outlet and a half internal angel of 45° an inner pipe flow occurs during emptying. This means according to Eurocode that the dimension shall be based on the pressure which occurs during filling. The horizontal and vertical pressure and the pressure made from the friction are calculated for the different cross sections.   The CAD software Pro Engineer and the finite element extension Mechanica is used to model the current silos and perform analysis for stress and buckling. The models have four symmetry planes therefore only one eighth of the current structure is modeled, corresponding to half a wall element and half a pole. The models are created as shell models and boundary conditions are applied in all symmetrical planes and on the top and bottom of the pole. The structure of the silos is simplified since no screws or radius is modeled. The pressure calculated according to Eurocode is converted into forces and applied to the models. The whole structure is modeled in steel with yield strength of 180 MPa.   The company’s older dimension standards are applied on the wall elements and analyzed. To investigate where to most critical areas for buckling occurs a buckling analysis based on a static analysis of the models is performed. The results from the buckling analysis for the silo wall element consisting of corrugated sheet with the width of 3.0 m shows that buckling occurs on the second bottom section at 72 % of the applied force. For the silo consisting of the same wall element but with the width of 2.5 m buckling occurs at the top section where the buckling force amounts to 62 % of the applied force. For the silos with wall elements consisting of plain sheet mounted on corrugated sheet buckling occurs at 3-4 % of the applied force for the two wall widths. Analysis show that the low values of buckling load on the plane sheet is a result from local buckling. In order to dimension the corrugated sheet to prevent it from buckling when the plane sheet is mounted a free body diagram is made for the corrugated sheet to obtain the acting forces. The buckling analysis of the corrugated sheet, with wall width 3.0 m, which is pressurized by the plane sheet shows that buckling occurs on the silos second bottom section. Buckling occurs at 59 % of the applied force for the silo with wall width of 2.5 m buckling occurs at 51 % of the applied force on the silo top section.   Since the goal is that the corrugated sheets are not to be subject to buckling, the thickness of the sheets is iterated until the buckling force is equal to at least 110 % of the applied force. This generates an increased thickness for the lower four sections for the silo with wall element consisting of corrugated sheet with wall width of 3.0 m. For the silo with the same wall elements but with a wall width of 2.5 m, the dimensions of the top two sections need to increase. Regarding the silos with wall elements consisting of plane sheet mounted on corrugated sheet an increase in dimension is needed for the corrugated sheet for the five lowest sections for the wall width of 3.0 m.   With a wall width of 2.5 m the same dimension can be used as when the silo wall elements consist of only corrugated sheet. If the plane sheet is not to be exposed for buckling the thickness of the sheets needs to be increased from between 5.5 mm and 1.5 mm. All calculations of the sheet dimensions are obtained by a conservative thinking which means that the company’s older dimensions may be correct. However, the resulting dimensions are reasonable for the corrugated sheets.

Summaries in English And Afrikaans / Text in Afrikaans / The biography focused on the life and work of Evert Philippus Groenewald in church historical perspective within the context of the twentieth century. It endeavoured to make a contribution to a period in the history of the Dutch Reformed Church (DRC) against the background of strife in matters related to the church, politics and other areas. Groenewald taught New Testament subjects as professor at the Faculty of Theology (Section 8 - DRC) at the University of Pretoria for 33 years. As one of the first four professors of the new Faculty of Theology he helped to lay a sound foundation. He served as dean of the Faculty for 21 years (1948 - 1969). As administrator and lecturer he played an important role in the establishment, recognition and development of the Faculty. He was a born exegete, the Nestor of New Testament science in SA. He is regarded as a great theologian and the doyen of New Testament research and exegesis in SA. As writer of Afrikaans theological literature Groenewald undeniably fulfilled the role of a pioneer. This was recognised by the award of the Stals-prize for Theology to him, the first time in history. Apart from this he received four honourary doctorates and various other important awards. He made an important contribution to Bible translation and translatory science. As church and cultural leader he strove to find solutions for the problems of his church and people. He made a significant contribution in the ecumenical field. Apartheid was given its biblical foundation by Groenewald during the forties. His intention was to stimulate thought in the search for a solution to the serious racial problem. Further facets of Groenewald's work await research in order to paint a representative theological portrait of him. / Die studie fokus op die lewe en werk van Evert Philippus Groenewald in kerkhistoriese perspektief binne die konteks van die twintigste eeu. Die klem het op die periode tussen die dertiger- en tagtigerjare geval. Daar is gepoog om 'n bydrae te lewer tot 'n tydperk in die geskiedenis van die NGK teen die agtergrond van die stryd wat op vele gebiede gevoer is. Die invloed van vroom ouers, 'n aantal ingrypende gebeurtenisse in sy lewe en die eise van die tyd het hom ge·inspireer. Hy was kind en teoloog van sy tyd, egter ook van sy kerk. Wat hy verwoord het was deel van die NGK se Calvinistiese en gereformeerde historiese erfenis en die NGK se beleid. Hy het sy eie teologiese stempel egter ongetwyfeld daarop afgedruk. Hy onderrig vir 33 jaar (1938 tot 1970) Nuwe Testamentiese vakke as professor aan die Teofogiese Fakulteit (Afd 8) van UP. As een van die eerste vier professore help hy om 'n stewige grondslag te le. Hy was vir 21 jaar (1948-1969) dekaan van die Fakulteit. In 1970 het hy geemeriteer. Hy het nie die akademie as sy enigste roeping beskou nie. Hy het sy kerk, volk en die gemeenskap gedien. Na sy aftrede lewer hy sy belangrikste bydraes tot Bybelvertaling, die NAV (gepubliseer in 1983), en die vertaalwetenskap. Hy het wat sy lewenstaak betref sy belangrikste bydrae as dosent gelewer. As administrateur het hy die Fakulteit help vestig en uitbou. Hy was 'n gebore eksegeet, die Nestor van die Nuwe Testamentiese wetenskap in SA. Hy word beskou as 'n belangrike teoloog, die doyen van die Nuwe Testamentiese navorsing en eksegese in SA. Die tradisie van die N T eksegese wat hy in die beoefening van die teologie gevestig het, kan as sy heel belangrikste bydrae beskou word. Hy word ook as volksteoloog getipeer. As skrywer het Groenewald 'n pioniersrol vervul vanwee sy groot bydrae tot die ontluikende Afrikaanse teologiese literatuur. Hy het die prestasie behaal dat die Stals-prys vir Teologie, die eerste keer in die geskiedenis, aan horn toegeken is. Hy is met vier eredoktorsgrade en verskeie ander belangrike toekennings vereer. As kerk- en kultuurleier het Groenewald se bekwaamhede sterk na vore gekom. Hy het die behoeftes van sy kerk en volk raakgesien en horn beywer om oplossings vir die ingewikkelde vraagstukke te vind. Die NGK, UP, vele instansies en die Staat het gebruik gemaak van sy talente. Apartheid is in die veertigerjare deur hom Bybels fundeer. Sy bedoeling was om teologiese denke te stimuleer in die soeke na 'n oplossing vir die ernstige rassevraagstuk. Hy het apartheid as teologies-praktiese oplossing aangebied. Hy het na 'n eerlike oplossing binne die destydse konteks gesoek wat regverdig teenoor die nieblanke sou wees en van die blanke groot offers sou vra. Die wyse waarop apartheid algaande toegepas is, het meegebring dat daar afgewyk is van wat Groenewald in 1947 beoog het. Dit het in 'n ideologiese monster ontaard wat groot pyn en lyding veroorsaak het. Groenewald was een van 'n klein aantal kerkfigure wat met hul dade in die twintigste eeu van stryd en worsteling die Suid-Afrikaanse kerkgeskiedenis verryk het. Sy bydrae is deur hierdie proefskrif slegs ten dele ontgin en verdere fasette van sy werk wag om nagevors te word ten einde uiteindelik 'n omvattende teologiese portret van hom te skilder. / Christian Spirituality, Church History and Missiology / Th. D. (Church History)

The thesis provide an analysis of environment that might influence processes of socialization of youth people. Next to this description of socially and/or pedagogically working organizations and institutions that focus on youth and their parents in the area of risk behaviour with an aim to prevent dependence on mind-altering substances is provided. Theoretical part of the thesis discusses socio-cultural factors. Therefore roles especially of family as primary institution responsible for socialization as well as of socio-pedagogically working organizations in drug prevention targeting youth and their parents is described. Empirical part of the thesis addresses ways that parents of young users of drugs used to prevent risky behaviour of their children, what sources and types of informations about drugs they used, and to what extend they have been prepared to solve potential problems that might result in drug dependence. Also with the use of a narrative analysis it is discussed where parents sought professional help and what was their opinion about quality of socio- pedagogical intervention delivered by individual organizations.