Cloning and expression of the Propionibacterium shermanii methylmalonyl-CoA epimerase gene in Escherichia coli : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Biochemistry at Massey University

Genomic DNA was isolated from Propionibacterium shermanii (52W). A 454 bp DNA fragment coding for the methylmalonyl-CoA epimerase (EC 5.1.99.1, subsequently referred to as epimerase) was amplified from genomic DNA by the polymerase chain reaction using primers designed from the known DNA sequence of the gene. The P. shermanii epimerase gene was ligated into the 2.47 kbp expression vector pT7-7. The ligation reaction mixture was transformed into electroporation competent E.coli XL1-Blue cells. Plasmid DNA prepared from several transformants was analysed, by agarose gel electrophoresis of restriction enzyme digestions, and transformed into E.coli SRP84/pGP1-2 cells to identify potential epimerase expression constructs (pTEEX) by heat shock induction. The insert DNA of one of the putative pTEEX epimerase constructs was fully sequenced and shown to be identical to the known DNA sequence of the epimerase gene described by Davis (1987). Using the sequenced expression construct pTEEX, recombinant epimerase was expressed to 20-35% of the total cell protein in the protease deficient E.coli strain SRP84 using the dual plasmid expression system of Tabor and Richardson (1985). The recombinant epimerase was ~95-100% soluble in E.coli. The recombinant epimerase and the 'wild-type' epimerase produced by P. shermanii were purified using the procedures developed for the 'wild-type' epimerase. The addition of a heat-treatment step (70°C for 15 min) early in the purification of the recombinant enzyme successfully exploited the unusually high thermostability of the epimerase protein. The epimerase protein was found to have an anomalously low electrophoretic mobility in a modified Laemmli discontinuous Tris-glycine alkaline buffer system for SDS-PAGE gels compared to the Weber and Osborn continuous phosphate buffer system. Using the latter system, a subunit molecular weight of 16.6 kDa was obtained. This is consistent with the molecular weight of 16.72 kDa (methionine on) calculated from the inferred amino acid sequence. The N-terminal sequence of the purified 'wild-type' and recombinant epimerases were identical although only half of N-terminal methionine residues were removed from the recombinant protein. The subunit molecular weight, specific activity, activation by divalent metal ions and behaviour in crystallization trials of the 'wild-type' and recombinant epimerases were very similar. Recombinant epimerase crystals were grown in a buffer containing 0.2 M ammonium acetate and 0.1 M citrate, pH 5.6, containing 30% PEG 4000 as precipitant. These crystals were relatively poorly ordered and diffracted to only 4.5 Ǻ resolution, but crystals of the recombinant epimerase that diffract to 2.6Ǻ can be grown under appropriate conditions

Development of a robust structural health monitoring system for wind turbine foundations

The construction of onshore wind turbines has rapidly been increasing as the UK attempts to meet its renewable energy targets. As the UK’s future energy depends more on wind farms, safety and security are critical to the success of this renewable energy source. Structural integrity is a critical element of this security of supply. With the stochastic nature of the load regime a bespoke low cost structural health monitoring system is required to monitor integrity. This paper presents an assessment of ‘embedded can’ style foundation failure modes in large onshore wind turbines and proposes a novel condition based monitoring solution to aid in early warning of failure

Conséquences de la diffusion des innovations technologiques sur l'emploi industriel en Tunisie : Une analyse par les données de panel

Working paper serie RMT (WPS 10-02), 26 p.The subject of this article, inspired by the compensation theory, is to discover the short and long term effect of technological innovations diffusion on employment in the case of developing countries, especially in Tunisia. Our results show that, even if in the short term, the effect of imported technological innovation and patents on employment is positive; this effect is negative in the medium and the long term. This result contradicts the theoretical predictions. This can be explained by the fact that Tunisian economy remains basically consumer and little productive of technological innovation. The complementarities between capital and job explain the increase of job opportunities with the increase of the imported technologies in short term. However, in medium and long term, Tunisian firms seem unable to put their technology in level. In most firms, the majority of inputs are imported and the use of obsolete machines and not much qualified employees is frequent. The process therefore does not implicate learning, nor massive training of technicians and engineers, and nor the production of similar technologies. When firms take advantage of the new technologies, the investors are encouraged to create more firms and new jobs. Moreover, the existence of the constraints of employees' mobility seems to prevent the functioning of the compensation mechanisms.L'objectif de cet article est, en s'inspirant de la théorie de compensation, de déterminer les effets, de court et de long terme, de la diffusion des innovations technologiques sur l'emploi pour le cas d'un pays en développement (en l'occurrence la Tunisie). Nos résultats montrent que, si l'effet à court terme des innovations technologiques importées et des brevets sur l'emploi est positif, cet effet est négatif à moyen et long terme. Ce résultat contredit les prédictions théoriques. Cette contradiction peut être expliquée par le fait que l'économie tunisienne demeure fondamentalement consommatrice et encore très peu productrice des innovations technologiques. La complémentarité entre capital et travail explique ainsi l'augmentation de court terme de l'emploi suite à l'augmentation des importations de technologies. Néanmoins, à moyen et long terme les entreprises tunisiennes semblent ne pas pouvoir mettre leur technologie à niveau. Dans la plupart des entreprises, la majorité des inputs sont importés employant des machines souvent déjà obsolètes et une main d'oeuvre très peu qualifiée. Le processus n'implique donc pas l'apprentissage, ni la formation massive de cadres ou de techniciens, ni la production de technologies similaires. Quand les firmes peuvent tirer parti des nouvelles technologies, les investisseurs sont encouragés à créer des entreprises et des emplois nouveaux. S'ajoute l'existence de contraintes à la mobilité de la main-d'oeuvre. Ces contraintes semblent avoir empêcher le bon fonctionnement des mécanismes de compensatio

Does Rosetta Stone Declare the Death of the Teacher?

Technology ushers in fundamental structural changes that can be essential to achieve significant improvements in all sectors including education This justifies the great attention that many universities have paid to the incorporation of technology into the classroom in the recent years The faculty of Science and Arts in Khulais Jeddah Saudi Arabia and in an attempt to keep up with the technological developments adopted a computer- mediated software called Rosetta Stone The latter is claimed to perform the same roles as the teacher and yield effective language learning outcomes from the part of learners Indeed this paper addressed such claims by identifying the teachers perceptions towards technology in general and Rosetta Stone in particular It also aimed to explore the teachers attitudes towards the potential adjustments they may make to their roles after the introduction of Rosetta Stone into the educational landscape and whether such programs can replace them Data was gathered via a semi- structured interview and a questionnaire The results showed that despite their highly favorable opinions towards technology teachers emphasized the importance of selecting the kind of technology to be employed in the classroom according to its suitability Findings also mirrored the teachers views that Rosetta Stone is no substitute for the teacher This implies that technology cannot set teachers aside or take them over by contrast it adds to their roles This paper includes more implications of the findings which urge the need for more researches to be carried out so as a better understanding of the classroom environment is achieved which will in turn guarantee better learning attainment

caractéristiques notables du système tunisien d’innovation

Dans ce papier, nous analysons le système tunisien d’innovation en mobilisant la méthodologie de l’approche en termes de systèmes sociaux d’innovation et de production développée par Amable, Barré et Boyer (1997). Pour ce faire, nous avons présenté dans un premier temps les inputs, ainsi que les outputs, de la recherche scientifique et des activités d’innovation. Dans un deuxième temps, en mettant l’accent sur les interrelations entre les différentes composantes du système d’innovation, nous avons essayé d’analyser les forces et les faiblesses de ce système. Notre analyse montre qu’il existe deux pôles industriels en situations extrêmes du point de vue de la recherche et de l’innovation, d’une part le secteur agroalimentaire, d’autre part le textile et l’habillement. En dépit des mesures prises par l’Etat, l’articulation entre la production de connaissances et leur application reste insuffisante. Dans ce cadre d’idées, nous avons particulièrement mis l’accent sur la difficile coordination entre la politique éducative et la politique industrielle. In this paper, we analyze the Tunisian system of innovation by mobilizing the methodology of the social systems of innovation and production approach, developed by Amable, Barré and Boyer. First, we present the inputs, as well as the outputs, of scientific research and innovation activities. Secondly, by stressing the interrelationships between the various components of the system of innovation, we try to analyze the forces and the weaknesses of this system. Our analysis shows that there are two industrial poles in extreme situations from the point of view of research and innovation activities: on the one hand the agrofood sector, on the other hand the textile and clothing sector. In spite of the policies implemented by the State, the articulation between the production of knowledge and its application remains insufficient. Within this framework of ideas, we particularly stress the difficult coordination between the educational policy and the industrial policy.system of innovation, Tunisian, industrial policy, educational policy

Conséquences de la diffusion des innovations technologiques sur l'emploi industriel en Tunisie : Une analyse par les données de panel

L'objectif de cet article est, en s'inspirant de la théorie de compensation, de déterminer les effets, de court et de long terme, de la diffusion des innovations technologiques sur l'emploi pour le cas d'un pays en développement (en l'occurrence la Tunisie). Nos résultats montrent que, si l'effet à court terme des innovations technologiques importées et des brevets sur l'emploi est positif, cet effet est négatif à moyen et long terme. Ce résultat contredit les prédictions théoriques. Cette contradiction peut être expliquée par le fait que l'économie tunisienne demeure fondamentalement consommatrice et encore très peu productrice des innovations technologiques. La complémentarité entre capital et travail explique ainsi l'augmentation de court terme de l'emploi suite à l'augmentation des importations de technologies. Néanmoins, à moyen et long terme les entreprises tunisiennes semblent ne pas pouvoir mettre leur technologie à niveau. Dans la plupart des entreprises, la majorité des inputs sont importés employant des machines souvent déjà obsolètes et une main d'oeuvre très peu qualifiée. Le processus n'implique donc pas l'apprentissage, ni la formation massive de cadres ou de techniciens, ni la production de technologies similaires. Quand les firmes peuvent tirer parti des nouvelles technologies, les investisseurs sont encouragés à créer des entreprises et des emplois nouveaux. S'ajoute l'existence de contraintes à la mobilité de la main-d'oeuvre. Ces contraintes semblent avoir empêcher le bon fonctionnement des mécanismes de compensation

Innovations technologiques, mobilité et demande de main-d’oeuvre qualifiée. Une analyse des industries tunisiennes (Technological innovations, mobility and skilled-labour deamnd : an analysis of tunisian industries).

L’objectif de cet article est de déterminer les effets de la diffusion des innovations technologiques sur la demande de la main d'oeuvre qualifiée pour le cas d’un pays en développement (en l’occurrence la Tunisie). Cette étude présente trois originalités. La première tient à la prise en compte des coûts d'ajustement. La deuxième originalité tient à la mesure de l’innovation. La troisième originalité tient à la nature de données utilisées. L'étude économétrique montre l'existence d'un biais technologique, qui favorise la demande des cadres -supposés la main-d'oeuvre la plus qualifiée-par les industries. A cet effet s'ajoute un second effet favorable à la main d'oeuvre qualifiée: cette dernière est plus fortement complémentaire au capital que les ouvriers, qui sont supposés la main-d'oeuvre non qualifiée.The aim of this article is to discover the effect of technological innovations diffusion on skilled-labour demand in the case of developing countries (especially in Tunisia). This research presents three originalities. The first takes into considerations the adjustment costs. The second originality depends on the innovation measurement. The third originality is due to the nature of used data. The econometric research shows the existence of skill-biased technical change that favour the industries demand of managers. These later are supposed the most skilled-labour. In addition to this effect, they complement the capital rather than the manual workers, who are supposed unskilled-labour.

Settlement rehabilitation of a 35 year old building : case study integrated with analysis and implementation

This paper presents a rehabilitation project concerning the settlement of a 35 year old building. The foundation system of the northwest wing of the building consists of strip footings and slabon-grade. Differential settlement results in significant cracking of the masonry partition walls located on the footing and hence rehabilitation of the footing is required to stabilize the foundation system. Geotechnical and structural investigations are conducted, including site borings and analytical modeling based on one-dimensional consolidation theory that is incorporated into a finite element analysis. The predictive model exhibits that the differential settlement does not cause noticeable distress for the primary structural members, whereas the continued settlement affects use of the building. Site implementation is performed with the pushpile method to terminate the continuous settlement of the foundation

El impacto económico de la guerra Rusia y Ucrania

El trabajo trata de como ha sido el impacto de la guerra entre Rusia y Ucrania sobre la economía europea y rusa. Una guerra entre dos países que son importantes exportadores de productos primarios en el mundo, energéticos para Rusia y agroalimentarios para Ucrania, es de gran relevancia global. Cada día puede surgir un evento que lleve a un cambio en el comercio mundial y las relaciones entre estos países y el resto del mundoMáster en Comercio Exterio

Wind turbine foundation deburial sensors based on induction-heated ceramic patches

The deburial and scouring of concrete wind turbine and bridge foundations presents a risk to structural stability and safety. In this work, we present a novel, ceramic temperature-sensing patch which can detect whether sections of a foundation are buried or exposed to air. The sensor patches, applied to concrete specimens, were fabricated by loading a geopolymer with 0 - 60 wt% ground magnetite. The magnetite content allowed the patches to be heated using an induction coil, while temperature profiles were monitored via changes in patch electrical impedance. Sensor patches were left uncoated, or were coated in surface-water, soil and sand. Each material provided a unique thermal signature which, with simple signal processing, could be used to reliably detect whether the patch was buried