Hegemony, Private Actors, and International Institutions: Transnational Corporations as the agents of transformation of the trade regime from GATT to the WTO

Diese Dissertation hat das Ziel, zum einen die Transformation des Handelssystems von der GATT zur Welthandelsorganisation (WTO) im Kontext einer veränderten Weltordnung und zum anderen die Rollen von transnationalen Unternehmen im Rahmen dieser Transformation zu untersuchen und zu verstehen. Die Arbeit wird theoretisch vom Neogramscianismus angeleitet, da die etablierten Ansätzen in den Internationalen Beziehungen und der International Politischen Ökonomie nur unzureichend die intersubjektive Natur von Regimen und nicht-staatlichen Akteuren darstellen. Für Anhänger des Neogramscianismus sind internationale Regime intersubjektive Einheiten, deren Zusammenspiel von Ideen und Machtkonfigurationen historische Strukturen prägen. Die Hegemonie ist ein Konzept, das soziale Einflüsse als Agenten historischen Wandels in international Regimen und der Weltordnung zusammenbindet. Mit dem Konzept der Hegemonie wird eine Machtsituation beschrieben, in der politische Macht in legitime Autorität übersetzt wird, indem die Zustimmung subalterner Akteure eingeholt wird. Hegemonie beinhaltet die konsensuellen Aspekte von Machtausübung in einer jeweiligen Weltordnung. Diese Dissertation argumentiert vor allem, dass die Transformation des Handelssystems als hegemonisch bezeichnet werden kann, da sie parallel mit der Transformation der Weltordnung von einer von den USA dominierten Nachkriegszeit zu einer neoliberalen Hegemonie stattfand. Mit der Transformation zur Welthandlungsorganisation wird der legale Rahmen des Handelssystems neu strukturiert und ihre normative Grundlagen neu definiert, wodurch der ethische Rahmen des Neoliberalismus reflektiert wird. Diese Änderungen werden in der neuartigen Anerkennung der legitimen Autorität des Marktes gegenüber Nationalstaaten und der Anerkennung von der Notwendigkeit von bindenden Disziplinen, die Regierungen übergeordnet sind, reflektiert. Diese Dissertation analysiert zwei Fälle, um die Rolle von transnationalen Unternehmen innerhalb diese Transformationsprozesses zu erklären. Dabei wird der Fokus vor allem auf die Aktivitäten und Fähigkeiten der Unternehmen gerichtet, die Ausrichtung des Handelsregimes zu bestimmen. Die erste Studie untersucht die Eingliederung von Dienstleistungen in das GATT Regime vor und während der Uruguay-Runde (1986 – 1994) und argumentiert, dass diese Eingliederung zu einer Neudefinierung von Liberalisierung und Normen der Nichtdiskriminierung führte. Die zweite Studie analysiert den gescheiterten Versuch, ausländische Direktinvestitionen noch bevor und während der 2001 begonnenen Doha Runde in die Welthandelsorganisation zu integrieren. Letztendlich wird in dieser Dissertation argumentiert, dass transnationale Unternehmen, die in den Vereinigten Staaten ansässig sind, hegemonische Agenten der Regimetransformation waren und eine wichtige Rolle dabei gespielt haben, Dienstleistungen in das GATT einzubinden. Und zwar gelang ihnen dies durch eine in den späten 1970er Jahren begonnenen Kampagne. Auf der einen Seite war die Kampagne darin erfolgreich, etablierte Denkstrukturen zu Handelsthemen systematisch im Sinne des Neoliberalismus zu verändern – und zwar sowohl hinsichtlich der normativen Inhalte als auch der intersubjektiven Bedeutungen des Regimes. Auf der anderen Seite deutet der Fall des Investitionsabkommens die Grenzen der hegemonischen Ideen, Institutionen, und Strömungen seit den frühen 90er Jahren an. Transnationale Unternehmen, die in Europa ansässig waren, sind mit ihren Bemühungen gescheitert, das Regime weiter zu transformieren und das Thema Investitionen in die legalen und normativen Rahmenbedingungen der WTO zu integrieren. Die Prioritäten und Strategien der transnationalen Unternehmen, die Agenda der WTO zu beeinflussen, waren beschränkt und wurden im Kontext einer angefochtenen neoliberalen Hegemonie geformt, die wiederum von dem Widerstand und anti-hegemonischen Kampagnen der Zivilgesellschaft beeinflusst wurden. Die Analyse in dieser Dissertation wurde durch eine qualitative Diskursanalyse von Sekundär- und Primärquellen durchgeführt: Regierungsvorschläge, Verhandlungstexte, Konferenzzusammenfassungen und Statements von Unternehmen.

Accuracy of outcome anticipation, but not gaze behavior, differs against left- and right-handed penalties in team-handball goalkeeping

Low perceptual familiarity with relatively rarer left-handed as opposed to more common right-handed individuals may result in athletes' poorer ability to anticipate the former's action intentions. Part of such left-right asymmetry in visual anticipation could be due to an inefficient gaze strategy during confrontation with left-handed individuals. To exemplify, observers may not mirror their gaze when viewing left- vs. right-handed actions but preferentially fixate on an opponent's right body side, irrespective of an opponent's handedness, owing to the predominant exposure to right-handed actions. So far empirical verification of such assumption, however, is lacking. Here we report on an experiment where team-handball goalkeepers' and non-goalkeepers' gaze behavior was recorded while they predicted throw direction of left- and right-handed 7-m penalties shown as videos on a computer monitor. As expected, goalkeepers were considerably more accurate than non-goalkeepers and prediction was better against right- than left-handed penalties. However, there was no indication of differences in gaze measures (i.e., number of fixations, overall and final fixation duration, time-course of horizontal or vertical fixation deviation) as a function of skill group or the penalty-takers' handedness. Findings suggest that inferior anticipation of left-handed compared to right-handed individuals' action intentions may not be associated with misalignment in gaze behavior. Rather, albeit looking similarly, accuracy differences could be due to observers' differential ability of picking up and interpreting the visual information provided by left- vs. right-handed movements.

HelF, a suppressor of RNAi mediated gene silencing in Dictyostelium discoideum

RNA interference (RNAi) is a recently discovered process, in which double stranded RNA (dsRNA) triggers the homology-dependant degradation of cognate messenger RNA (mRNA). In a search for new components of the RNAi machinery in Dictyostelium, a new gene was identified, which was called helF. HelF is a putative RNA helicase, which shows a high homology to the helicase domain of Dicer, to the helicase domain of Dictyostelium RdRP and to the C. elegans gene drh-1, that codes for a dicer related DExH-box RNA helicase, which is required for RNAi. The aim of the present Ph.D. work was to investigate the role of HelF in PTGS, either induced by RNAi or asRNA. A genomic disruption of the helF gene was performed, which resulted in a distinct mutant morphology in late development. The cellular localization of the protein was elucidated by creating a HelF-GFP fusion protein, which was found to be localized in speckles in the nucleus. The involvement of HelF in the RNAi mechanism was studied. For this purpose, RNAi was induced by transformation of RNAi hairpin constructs against four endogenous genes in wild type and HelF- cells. The silencing efficiency was strongly enhanced in the HelF K.O. strain in comparison with the wild type. One gene, which could not be silenced in the wild type background, was successfully silenced in HelF-. When the helF gene was disrupted in a secondary transformation in a non-silenced strain, the silencing efficiency was strongly improved, a phenomenon named here “retrosilencing”. Transcriptional run-on experiments revealed that the enhanced gene silencing in HelF- was a posttranscriptional event, and that the silencing efficiency depended on the transcription levels of hairpin RNAs. In HelF-, the threshold level of hairpin transcription required for efficient silencing was dramatically lowered. The RNAi-mediated silencing was accompanied by the production of siRNAs; however, their amount did not depend on the level of hairpin transcription. These results indicated that HelF is a natural suppressor of RNAi in Dictyostelium. In contrast, asRNA mediated gene silencing was not enhanced in the HelF K.O, as shown for three tested genes. These results confirmed previous observations (H. Martens and W. Nellen, unpublished) that although similar, RNAi and asRNA mediated gene silencing mechanisms differ in their requirements for specific proteins. In order to characterize the function of the HelF protein on a molecular level and to study its interactions with other RNAi components, in vitro experiments were performed. Besides the DEAH-helicase domain, HelF contains a double-stranded RNA binding domain (dsRBD) at its N-terminus, which showed high similarity to the dsRBD domain of Dicer A from Dictyostelium. The ability of the recombinant dsRBDs from HelF and Dicer A to bind dsRNA was examined and compared. It was shown by gel-shift assays that both HelF-dsRBD and Dicer-dsRBD could bind directly to long dsRNAs. However, HelF-dsRBD bound more efficiently to dsRNA with imperfect matches than to perfect dsRNA. Both dsRBDs bound specifically to a pre-miRNA substrate (pre-let-7). The results suggested that most probably there were two binding sites for the proteins on the pre-miRNA substrate. Moreover, it was shown that HelF-dsRBD and Dicer-dsRBD have siRNA-binding activity. The affinities of the two dsRBDs to the pre-let-7 substrate were also examined by plasmon surface resonance analyses, which revealed a 9-fold higher binding affinity of the Dicer-dsRBD to pre-let-7 compared to that of the HelF-dsRBD. The binding of HelF-dsRBD to the pre-let-7 was impaired in the presence of Mg2+, while the Dicer-dsRBD interaction with pre-let-7 was not influenced by the presence of Mg2+. The results obtained in this thesis can be used to postulate a model for HelF function. In this, HelF acts as a nuclear suppressor of RNAi in wild type cells by recognition and binding of dsRNA substrates. The protein might act as a surveillance system to avoid RNAi initiation by fortuitous dsRNA formation or low abundance of dsRNA trigger. If the protein acts as an RNA helicase, it could unwind fold-back structures in the nucleus and thus lead to decreased RNAi efficiency. A knock-out of HelF would result in initiation of the RNAi pathway even by low levels of dsRNA. The exact molecular function of the protein in the RNAi mechanism still has to be elucidated. RNA interferenz (RNAi) ist ein in jüngster Zeit entdeckter Mechanismus, bei dem doppelsträngige RNA Moleküle (dsRNA) eine Homologie-abhängige Degradation einer verwandten messenger-RNA (mRNA) auslösen. Auf der Suche nach neuen Komponenten der RNAi-Maschinerie in Dictyostelium konnte ein neues Gen (helF) identifiziert werden. HelF ist eine putative RNA-Helikase mit einer hohen Homologie zur Helikasedomäne der bekannten Dicerproteine, der Helikasedomäne der Dictyostelium RdRP und zu dem C. elegans Gen drh-1, welches für eine Dicer-bezogene DExH-box RNA Helikase codiert, die am RNAi-Mechanismus beteiligt ist. Das Ziel dieser Arbeit war es, die Funktion von HelF im Zusammenhang des RNAi oder asRNA induzierten PTGS zu untersuchen. Es wurde eine Unterbrechung des helF-Gens auf genomischer Ebene (K.O.) vorgenommen, was bei den Mutanten zu einer veränderten Morphologie in der späten Entwicklung führte. Die Lokalisation des Proteins in der Zelle konnte mit Hilfe einer GFP-Fusion analysiert werden und kleinen Bereichen innerhalb des Nukleus zugewiesen werden. Im Weiteren wurde der Einfluss von HelF auf den RNAi-Mechanismus untersucht. Zu diesem Zweck wurde RNAi durch Einbringen von RNAi Hairpin-Konstrukten gegen vier endogene Gene im Wiltypstamm und der HelF--Mutante induziert. Im Vergleich zum Wildtypstamm konnte im HelF--Mutantenstamm eine stark erhöhte „Silencing“-Effizienz nachgewiesen werden. Ein Gen, welches nach RNAi Initiation im Wildtypstamm unverändert blieb, konnte im HelF--Mutantenstamm erfolgreich stillgelegt werden. Durch sekundäres Einführen einer Gendisruption im helF-Locus in einen Stamm, in welchem ein Gen nicht stillgelegt werden konnte, wurde die Effizienz des Stilllegens deutlich erhöht. Dieses Phänomen wurde hier erstmals als „Retrosilencing“ beschrieben. Mit Hilfe von transkriptionellen run-on Experimenten konnte belegt werden, dass es sich bei dieser erhöhten Stilllegungseffizienz um ein posttranskriptionelles Ereignis handelte, wobei die Stillegungseffizienz von der Transkriptionsstärke der Hairpin RNAs abhängt. Für die HelF--Mutanten konnte gezeigt werden, dass der Schwellenwert zum Auslösen eines effizienten Stillegens dramatisch abgesenkt war. Obwohl die RNAi-vermittelte Genstilllegung immer mit der Produktion von siRNAs einhergeht, war die Menge der siRNAs nicht abhängig von dem Expressionsniveau des Hairpin-Konstruktes. Diese Ergebnisse legen nahe, dass es sich bei der HelF um einen natürlichen Suppressor des RNAi-Mechanismus in Dictyostelium handelt. Im Gegensatz hierzu war die as-vermittelte Stilllegung von drei untersuchten Genen im HelF-K.O. im Vergleich zum Wildyp unverändert. Diese Ergebnisse bestätigten frühere Beobachtungen (H. Martens und W. Nellen, unveröffentlicht), wonach die Mechanismen für RNAi und asRNA-vermittelte Genstilllegung unterschiedliche spezifische Proteine benötigen. Um die Funktion des HelF-Proteins auf der molekularen Ebene genauer zu charakterisieren und die Interaktion mit anderen RNAi-Komponenten zu untersuchen, wurden in vitro Versuche durchgeführt. Das HelF-Protein enthält, neben der DEAH-Helikase-Domäne eine N-terminale Doppelstrang RNA bindende Domäne (dsRBD) mit einer hohen Ähnlichkeit zu der dsRBD des Dicer A aus Dictyostelium. Die dsRNA-Bindungsaktivität der beiden dsRBDs aus HelF und Dicer A wurde analysiert und verglichen. Es konnte mithilfe von Gel-Retardationsanalysen gezeigt werden, dass sowohl HelF-dsRBD als auch Dicer-dsRBD direkt an lange dsRNAs binden können. Hierbei zeigte sich, dass die HelF-dsRBD eine höhere Affinität zu einem imperfekten RNA-Doppelstrang besitzt, als zu einer perfekt gepaarten dsRNA. Für beide dsRBDs konnte eine spezifische Bindung an ein pre-miRNA Substrat nachgewiesen werden (pre-let-7). Dieses Ergebnis legt nah, dass es zwei Bindestellen für die Proteine auf dem pre-miRNA Substrat gibt. Überdies hinaus konnte gezeigt werden, dass die dsRBDs beider Proteine eine siRNA bindende Aktivität besitzen. Die Affinität beider dsRBDs an das pre-let-7 Substrat wurde weiterhin mit Hilfe der Plasmon Oberflächen Resonanz untersucht. Hierbei konnte eine 9-fach höhere Bindeaffinität der Dicer-dsRBD im Vergleich zur HelF-dsRBD nachgewiesen werden. Während die Bindung der HelF-dsRBD an das pre-let-7 durch die Anwesenheit von Mg2+ beeinträchtigt war, zeigte sich kein Einfluß von Mg2+ auf das Bindeverhalten der Dicer-dsRBD. Mit Hilfe der in dieser Arbeit gewonnen Ergebnisse lässt sich ein Model für die Funktion von HelF postulieren. In diesem Model wirkt HelF durch Erkennen und Binden von dsRNA Substraten als Suppressor von der RNAi im Kern. Das Protein kann als Überwachungsystem gegen eine irrtümliche Auslösung von RNAi wirken, die durch zufällige dsRNA Faltungen oder eine zu geringe Häufigkeit der siRNAs hervorgerufen sein könnte. Falls das Protein eine Helikase-Aktivität besitzt, könnte es rückgefaltete RNA Strukturen im Kern auflösen, was sich in einer verringerten RNAi-Effizienz wiederspiegelt. Durch Ausschalten des helF-Gens würde nach diesem Modell eine erfolgreiche Auslösung von RNAi schon bei sehr geringer Mengen an dsRNA möglich werden. Das Modell erlaubt, die exakte molekulare Funktion des HelF-Proteins im RNAi-Mechanismus weiter zu untersuchen.

Nuclear organisation and epigenetic regulation of gene expression in Dictyostelium discoideum

A series of vectors for the over-expression of tagged proteins in Dictyostelium were designed, constructed and tested. These vectors allow the addition of an N- or C-terminal tag (GFP, RFP, 3xFLAG, 3xHA, 6xMYC and TAP) with an optimized polylinker sequence and no additional amino acid residues at the N or C terminus. Different selectable markers (Blasticidin and gentamicin) are available as well as an extra chromosomal version; these allow copy number and thus expression level to be controlled, as well as allowing for more options with regard to complementation, co- and super-transformation. Finally, the vectors share standardized cloning sites, allowing a gene of interest to be easily transfered between the different versions of the vectors as experimental requirements evolve. The organisation and dynamics of the Dictyostelium nucleus during the cell cycle was investigated. The centromeric histone H3 (CenH3) variant serves to target the kinetochore to the centromeres and thus ensures correct chromosome segregation during mitosis and meiosis. A number of Dictyostelium histone H3-domain containing proteins as GFP-tagged fusions were expressed and it was found that one of them functions as CenH3 in this species. Like CenH3 from some other species, Dictyostelium CenH3 has an extended N-terminal domain with no similarity to any other known proteins. The targeting domain, comprising α-helix 2 and loop 1 of the histone fold is required for targeting CenH3 to centromeres. Compared to the targeting domain of other known and putative CenH3 species, Dictyostelium CenH3 has a shorter loop 1 region. The localisation of a variety of histone modifications and histone modifying enzymes was examined. Using fluorescence in situ hybridisation (FISH) and CenH3 chromatin-immunoprecipitation (ChIP) it was shown that the six telocentric centromeres contain all of the DIRS-1 and most of the DDT-A and skipper transposons. During interphase the centromeres remain attached to the centrosome resulting in a single CenH3 cluster which also contains the putative histone H3K9 methyltransferase SuvA, H3K9me3 and HP1 (heterochromatin protein 1). Except for the centromere cluster and a number of small foci at the nuclear periphery opposite the centromeres, the rest of the nucleus is largely devoid of transposons and heterochromatin associated histone modifications. At least some of the small foci correspond to the distal telomeres, suggesting that the chromosomes are organised in a Rabl-like manner. It was found that in contrast to metazoans, loading of CenH3 onto Dictyostelium centromeres occurs in late G2 phase. Transformation of Dictyostelium with vectors carrying the G418 resistance cassette typically results in the vector integrating into the genome in one or a few tandem arrays of approximately a hundred copies. In contrast, plasmids containing a Blasticidin resistance cassette integrate as single or a few copies. The behaviour of transgenes in the nucleus was examined by FISH, and it was found that low copy transgenes show apparently random distribution within the nucleus, while transgenes with more than approximately 10 copies cluster at or immediately adjacent to the centromeres in interphase cells regardless of the actual integration site along the chromosome. During mitosis the transgenes show centromere-like behaviour, and ChIP experiments show that transgenes contain the heterochromatin marker H3K9me2 and the centromeric histone variant H3v1. This clustering, and centromere-like behaviour was not observed on extrachromosomal transgenes, nor on a line where the transgene had integrated into the extrachromosomal rDNA palindrome. This suggests that it is the repetitive nature of the transgenes that causes the centromere-like behaviour. A Dictyostelium homolog of DET1, a protein largely restricted to multicellular eukaryotes where it has a role in developmental regulation was identified. As in other species Dictyostelium DET1 is nuclear localised. In ChIP experiments DET1 was found to bind the promoters of a number of developmentally regulated loci. In contrast to other species where it is an essential protein, loss of DET1 is not lethal in Dictyostelium, although viability is greatly reduced. Loss of DET1 results in delayed and abnormal development with enlarged aggregation territories. Mutant slugs displayed apparent cell type patterning with a bias towards pre-stalk cell types.

A Study of a Comprehensive Career Services System in Chinese Universities

Since 1999, with the adoption of expansion policy in higher education by the Chinese government, enrollment and graduate numbers have been increasing at an unprecedented speed. Accustomed to a system in which university graduates were placed, many students are not trained in “selling themselves”, which exacerbates the situation leading to a skyrocketing unemployment rate among new graduates. The idea of emphasizing career services comes with increasing employment pressure among university graduates in recent years. The 1998 “Higher Education Act” made it a legislative requirement. Thereafter, the Ministry of Education issued a series of documents in order to promote the development of career services. All higher education institutions are required to set up special career service centers and to set a ratio of 1:500 between career staff and the total number of students. Related career management courses, especially career planning classes, are required to be clearly included as specific modules into the teaching plan with a requirement of no less than 38 sessions in one semester at all universities. Developing career services in higher education has thus become a hot issue. One of the more notable trends in higher education in recent years has been the transformation of university career service centers from merely being the coordinators of on-campus placement into full service centers for international career development. The traditional core of career services in higher education had been built around guidance, information and placements (Watts, 1997). This core was still in place, but the role of higher education career services has changed considerably in recent years and the nature of each part is being transformed (Watts, 1997). Most services are undertaking a range of additional activities, and the career guidance issue is emphasized much more than before. Career management courses, especially career planning classes, are given special focus in developing career services in the Chinese case. This links career services clearly and directly with the course provision function. In China, most career service centers are engaging in the transformation period from a “management-oriented” organization to a “service-oriented” organization. Besides guidance services, information services and placement activities, there is a need to blend them together with the new additional teaching function, which follows the general trend as regulated by the government. The role of career services has been expanding and this has brought more challenges to its development in Chinese higher education. Chinese universities still remain in the period of exploration and establishment in developing their own career services. In the face of the new situation, it is very important and meaningful to explore and establish a comprehensive career services system to address student needs in the universities. A key part in developing this system is the introduction of career courses and delivering related career management skills to the students. So there is the need to restructure the career service sectors within the Chinese universities in general. The career service centers will operate as a hub and function as a spoke in the wheel of this model system, providing support and information to staff located in individual teaching departments who are responsible for the delivery of career education, information, advice and guidance. The career service centers will also provide training and career planning classes. The purpose of establishing a comprehensive career services system is to provide a strong base for student career development. The students can prepare themselves well in psychology, ideology and ability before employment with the assistance of effective career services. To conclude, according to the different characteristics and needs of students, there will be appropriate services and guidance in different stages and different ways. In other words, related career services and career guidance activities would be started for newly enrolled freshmen and continue throughout their whole university process. For the operation of a comprehensive services system, there is a need for strong support by the government in the form of macro-control and policy guarantee, but support by the government in the form of macro-control and policy guarantee, but also a need for close cooperation with the academic administration and faculties to be actively involved in career planning and employment programs. As an integral function within the universities, career services must develop and maintain productive relationships with relevant campus offices and key stakeholders both within the universities and externally.