La reproduction sexuée repose sur deux processus fondamentaux : la méiose qui permet la formation des gamètes dont le génome est haploïde et la syngamie qui permet, après fécondation, de restaurer la diploïdie par fusion des deux noyaux parentaux haploïdes. Alors que la méiose repose respectivement sur le génome maternel pour l'ovocyte et paternel pour le spermatozoïde, la restauration de la diploïdie dans le zygote repose exclusivement sur le génome maternel. Si un pronucleus maternel compétent pour la réplication est formé au terme de la méiose ovocytaire, le génome paternel quant à lui, n'acquiert cette compétence que sous l'influence de facteurs maternels. En effet, à la fin de la méiose, le génome paternel est « empaqueté » avec des protamines qui le rendent inactif pour toute fonction biologique, en particulier la réplication. L'éviction des protamines et leur remplacement par des histones maternelles sont des étapes indispensables à l'acquisition par le génome paternel de sa compétence à la réplication, préalable à la syngamie. Tous ces événements doivent être extrêmement coordonnés afin de permettre à un premier noyau zygotique comportant les deux lots de chromosomes parentaux de se former et d'entrer dans le premier cycle mitotique.Notre laboratoire a identifié yemanuclein-alpha, aussi appelé yemanuclein (yem) dans un crible moléculaire pour des gènes exprimés spécifiquement dans la lignée germinale femelle, et son premier allèle muté yem1. Cette mutation ponctuelle (V478E) a été identifiée dans un crible génétique de « stérilité femelle ». Une descendance exceptionnelle observée chez les femelles yem1, présente la propriété inattendue d'être parthénogénétique. Cette propriété révèle un double défaut chez le mutant : dans le processus de méiose ovocytaire qui conduit à la formation d'un pronucleus maternel haploïde mais aussi dans la formation d'un pronucleus paternel compétent pour la syngamie. Mes travaux de thèse ont porté sur les deux aspects de la fonction de la Yemanucléine. En conjuguant des méthodes de génétique, de biochimie, et de biologie cellulaire, nous avons pu mettre en évidence des fonctions essentielles de la Yemanucléine dans les étapes initiales de la prophase méiotique de l'ovocyte de drosophile. Nous avons pu montrer que la Yemanucléine joue un rôle clé dans la recombinaison méiotique et plus particulièrement dans la fréquence et la cinétique d'apparition des cassures double brin. Son association au complexe synaptonémal et au complexe cohésine, tous deux connus comme étant nécessaires à la ségrégation chromosomique, est un élément clé de cette fonction.Outre cette fonction méiotique, la Yemanucléine, facteur maternel, est aussi requise pour l'assemblage de la chromatine du pronucleus paternel. Nous montrons dans ce manuscrit qu'elle joue ce rôle à travers son action dans un troisième complexe, en partenariat avec la protéine HIRA. Le complexe multiprotéique contenant la protéine HIRA est connu pour sa fonction de chaperon du variant de l'histone H3.3 et son rôle dans l'assemblage de la chromatine du pronucleus paternel. La Yemanucléine est le premier membre de la famille HPC2/UBN1 caractérisé. Son rôle dans l'assemblage des nucléosomes découplé de la réplication est décrit pour la première fois dans ce manuscrit. C'est aussi la première fois qu'une protéine spécifique de la reproduction est décrite pour son implication à deux étapes clés de ce processus. / Sexual reproduction relies on two key events: formation of cells with a haploid genome through meiosis and restoration of diploidy through syngamy in the zygote. Meiosis completion is supported exclusively by the maternal genome for the oocyte and the paternal genome for the sperm cell. In contrast diploidy restoration in the zygote is entirely dependent on maternal factors. At the end of meiosis the maternal pronucleus is competent for replication, whereas the paternal genome is packed with protamines. These proteins need to be removed in the zygote and replaced by maternally provided histones before the paternal genome acquires competence for replication, a prerequisite for syngamy. All these events must be highly coordinated to allow the first zygotic nucleus to form with the two sets of parental chromosomes and enter the first mitotic cycle. Our laboratory has identified yemanuclein-alpha, also called yemanuclein (yem) in a molecular screen for genes specifically expressed in the female germ line and its first mutant allele yem1, in a female sterile screen. The role played by yem not only in the meiotic process through which a haploid maternal pronucleus is formed but also in the zygotic process that makes a paternal pronucleus competent for syngamy, is underscored by the obtention of exceptional parthenogenetic progeny from yem1 mothers.My thesis work is precisely dedicated to the analysis of both aspects of Yemanuclein function: in the oocyte and the zygote. Using genetic, biochemical and cell biology methods we were able to uncover essential functions of Yemanuclein in early meiotic prophase in the Drosophila oocyte. Using yem1 allele (V478E), we could show its requirement for meiotic recombination especially for the frequency and timing of the double strand breaks formation. Yemanuclein association with two protein complexes, the Synaptonemal Complex (SC) and the Cohesin complex known to be required for proper chromosome segregation, supports these findings. Beyond its meiotic function, Yemanuclein is also required in the zygote for assembly of paternal pronucleus chromatin. This is achieved through a third complex that acts as histone H3.3 chaperone. In the present manuscript we identify Yemanuclein as a partner of HIRA in its role in H3.3 nucleosome assembly and deposition on the paternal pronucleus. Interestingly Yemanuclein is the first member of the HPC2/UBN1 protein family ever characterized. The role of Yem/ HPC2/ UBN1 in replication independent chromatin remodeling remained elusive until very recently. Our work is original in that it is the first to report on a role of one member of this family in oocyte meiosis and paternal chromatin assembly in the zygote.

Le syndrome de DiGeorge ou microdélétion 22q11.2, est la délétion chromosomique la plus fréquente chez les êtres humains. Cette délétion est liée à la recombinaison homologue non-allélique au cours de la méiose induisant la perte d’en moyenne 40 gènes. Les études de corrélation génotype/phénotype chez les patients ont révélé des différences phénotypiques entre individus et cela indépendamment de la taille des microdélétions. L’hypothèse de l’implication des mécanismes épigénétiques dans la variabilité phénotypique observée a été soulevée mais reste encore inexplorée. C’est dans ce contexte que nous nous intéressons à l’étude des mécanismes épigénétiques au cours du développement, dans cette pathologie à travers l’utilisation d’un modèle de cellules souches pluripotentes induites humaines (hiPSs). En particulier, nous avons ciblé nos travaux sur le rôle de la chaperonne d’histone HIRA dont le gène est localisé dans la région délétée. HIRA est impliquée dans la déposition du variant d’histone H3.3, une histone majeure dans le cerveau. Afin de comprendre l’implication de HIRA dans les manifestations neurologique du syndrome de DiGeorge et en particulier dans la schizophrénie, nous avons développé et optimisé un nouveau protocole pour la différenciation de cellules hiPSCs en progéniteurs neuronaux, neurones corticaux et neurones dopaminergiques. L’ensemble de ces travaux ouvre donc de nouvelles perspectives pour la modélisation d’un grand nombre de pathologies, et dans le contexte du laboratoire, pour l’exploration des mécanismes épigénétiques associés à la variabilité phénotypique dans différentes maladies génétiques. / The DiGeorge syndrome also known as 22q11.2 microdeletion syndrome, is the most common deletion in humans. This deletion is linked to a non-allelic homologous recombination that occurs during meiosis and involves sequences called LCRs for "Low Copy Repeats". Depending on the LCRs involved, different deletions are observed, inducing the loss of approximately 40 genes. The absence of genotype/phenotype correlation in patients and the phenotypical differences regardless of the size of the microdeletion suggests the involvement of additional parameter. The hypothesis of epigenetic changes associated with the onset or variability of symptoms has been suggested but never investigated. In order to tackle this question, we decided to focus our attention of the role of the HIRA histone chaperone encoded by a gene located in the 22q11.2-deleted region. HIRA is involved in the deposition of the H3.3 histone variant, one of the main histone in the brain. In order to determine whether HIRA is implicated in the neurological manifestations in DiGeorge patients and particularly in schizophrenia, we developed and optimized a new protocol for the direct differentiation of human induced pluripotent stem cell (hiPSCs) into neural progenitors, cortical and dopaminergic neurons. In parallel, we developed a new protocol for hiPSCs differentiation toward the skeletal muscle lineage and the production of multinucleated muscle fibers. Altogether, these results open new perspectives for the modeling of a large number of pathologies, and in the context of our laboratory, the exploration of epigenetic mechanisms associated with phenotypic variability in different genetic diseases.

Th. Etat--Anthropologie sociale et ethnologie--Paris--Ecole des hautes études en sciences sociales, 1998. / Les vol. 2 et 3 contiennent les textes des chants populaires en langue malgache avec la trad. française en regard. Bibliogr. p. 430-451 (t. 1).

La reproduction sexuée implique une différentiation extrême des gamètes qui s’accompagne de profonds remaniements des chromosomes parentaux. Au moment de la fécondation, ces chromosomes doivent être rendus compétents pour la formation du premier noyau zygotique. Au cours de ma thèse, j’ai étudié l’importance fonctionnelle de plusieurs voies moléculaires paternelles et maternelles participant à cette étape chez la drosophile. Le complexe HIRA est impliqué dans l’assemblage de nucléosomes dans le pronoyau mâle à la fécondation. J’ai décrit le rôle de HIRA et de son partenaire Yemanucléine-α dans cette voie. J’ai caractérisé plus finement ce complexe en étudiant son rôle somatique dans l’assemblage des nucléosomes et son implication dans la stabilité de l’hétérochromatine, améliorant notre compréhension des besoins biologiques qui conditionnent sa conservation et son évolution. Je me suis aussi intéressé à diverses situations affectant l’intégrité des chromosomes parentaux à la fécondation. (1) J’ai décrit les conséquences catastrophiques pour la méiose femelle de l’expression naturelle d’un transposon à travers l’étude d’un cas de dysgénésie hybride. (2) J’ai contribué à montrer que la protéine K81 est essentielle pour la protection des télomères dans les chromosomes paternels au cours de la spermatogénèse. (3) J’ai participé à caractériser les conséquences pour les chromosomes paternels de l’incompatibilité cytoplasmique induite par la bactérie Wolbachia. Ensemble, ces travaux soulignent les particularités des chromosomes parentaux à la fécondation et aident à cerner l’importance des voies maternelles et paternelles dans leur intégration dans le premier noyau du zygote / Sexual reproduction involves dramatic gamete differentiation and profound parental chromosomes remodelling. At fertilization, these chromosomes need to be rendered competent for the formation of the fist zygotic nucleus. I have studied the functional relevance of several paternal and maternal molecular pathways that participate during this process in Drosophila. The HIRA complex is required for nucleosome assembly in the male pronucleus at fertilization. I have further described the rôle of HIRA and its obligatory partner Yemanuclein-α during this step. I have characterized the somatic roles of this complex during nucleosome assembly and its involvment in heterochromatin stability, which gives us a better understanding of the biological needs that drive its conservation and evolution. I have also focused on several situations where parental chromosomes integrity at fertilization is compromised. (1) I have described a meiotic catastrophe associated with the natural expression of a transposon in the female germline during hybrid dysgenesis. (2) I have contributed to show that K81 is an essential protein for telomere protection in paternal chromosomes during spermiogenesis. (3) I have participated in the characterization of the chromosomal abnormalities associated with cytoplasmic incompatibility induced by Wolbachia. Together, these results underscore the specificities of parental chromosomes at fertilization and shed light into the importance of maternal and paternal pathways for their integration in the first zygotic nucleus

La reproduction sexuée implique une différentiation extrême des gamètes qui s'accompagne de profonds remaniements des chromosomes parentaux. Au moment de la fécondation, ces chromosomes doivent être rendus compétents pour la formation du premier noyau zygotique. Au cours de ma thèse, j'ai étudié l'importance fonctionnelle de plusieurs voies moléculaires paternelles et maternelles participant à cette étape chez la drosophile. Le complexe HIRA est impliqué dans l'assemblage de nucléosomes dans le pronoyau mâle à la fécondation. J'ai décrit le rôle de HIRA et de son partenaire Yemanucléine-α dans cette voie. J'ai caractérisé plus finement ce complexe en étudiant son rôle somatique dans l'assemblage des nucléosomes et son implication dans la stabilité de l'hétérochromatine, améliorant notre compréhension des besoins biologiques qui conditionnent sa conservation et son évolution. Je me suis aussi intéressé à diverses situations affectant l'intégrité des chromosomes parentaux à la fécondation. (1) J'ai décrit les conséquences catastrophiques pour la méiose femelle de l'expression naturelle d'un transposon à travers l'étude d'un cas de dysgénésie hybride. (2) J'ai contribué à montrer que la protéine K81 est essentielle pour la protection des télomères dans les chromosomes paternels au cours de la spermatogénèse. (3) J'ai participé à caractériser les conséquences pour les chromosomes paternels de l'incompatibilité cytoplasmique induite par la bactérie Wolbachia. Ensemble, ces travaux soulignent les particularités des chromosomes parentaux à la fécondation et aident à cerner l'importance des voies maternelles et paternelles dans leur intégration dans le premier noyau du zygote

No description available.

L'établissement de latence du virus de l'Herpès simplex 1 (HSV1) est contrôlé par les corps nucléaires PML (PML-NBs) mais leur implication exacte reste encore confuse. Une des caractéristiques majeures de la latence du virus est l'interaction entre le génome viral et les PML-NBs formant des structures nommées viral DNA-containing PML-NBs (vDCP-NBs). L'utilisation d'un modèle d'infection de fibroblastes primaires humains, qui reproduit la formation des vDCP-NBs, combinée à une approche par immuno-FISH, a permis de montrer que les vDCP-NBs contiennent l'histone H3.3 et ses chaperons, les complexes DAXX-ATRX et HIRA. La protéine HIRA a été également observé au sein des vDCP-NBs dans les neurones des ganglions trijumeaux de souris infectées par HSV1. Des expériences de ChIP-qPCR dans des cellules exprimant H3.3 ou H3.1 tagguées, nous a permis de déterminer que le génome viral est spécifiquement chromatinisé avec l'histone H3.3. La déplétion d'une seule protéine des complexes chaperons de H3.3 affecte légèrement l'incorporation de H3.3 dans les génomes viraux latents. Au contraire, l'absence de PML diminue significativement la chromatinisation H3.3 du génome HSV-1 latent sans remplacement par H3.1. Cette étude démontre une régulation épigénétique du génomes HSV1 latent par une chromatinisation dépendante de H3.3 impliquant les complexes chaperons DAXX-ATRX et HIRA. De plus, cette étude révèle un rôle majeur des PML-NBs dans la chromatinisation H3.3 des génomes HSV1 latent / Herpes simplex virus 1 (HSV-1) latency establishment is tightly controlled by PML nuclear bodies (PML-NBs) although their exact implication is still elusive. A hallmark of HSV-1 latency is the interaction between latent viral genomes and PML-NBs leading to the formation of viral DNA-containing PML-NBs (vDCP-NBs). Using a replication defective HSV-1 infected human primary fibroblast model reproducing the formation of vDCP-NBs, combined with an IF-FISH approach developed to detect latent HSV-1, we show that vDCP-NBs contain both histone H3.3 and its chaperone complexes, i.e. the DAXX/ATRX and the HIRA complex. HIRA was also detected co-localizing with vDCP-NBs present in trigeminal ganglia neurons from HSV-1 infected WT mice. ChIP-qPCR performed on fibroblasts stably expressing tagged H3.3 or H3.1 show that latent HSV1 genomes are chromatinized almost exclusively with H3.3. Depletion of single proteins from the H3.3 chaperone complexes only mildly affects H3.3 deposition on the latent HSV1 genome. In contrast, absence of PML significantly impacts on the chromatinization of the latent genomes with H3.3 without replacement with H3.1. Consequently, the study demonstrates a specific epigenetic regulation of latent HSV-1 through an H3.3-dependent HSV-1 chromatinization involving both H3.3 chaperones DAXX/ATRX and HIRA complexes. Additionally, the study reveals that PML-NBs are major actors of the latent HSV-1 H3.3 chromatinization through a PML-NBs/histone H3.3/H3.3 chaperones axis

The electronic control, instrumentation and communication hardware is becoming more and more compact and faster in operation due to the increased use of large scale integration of semiconductor devices operating at higher speeds. The use of VLSI circuit based systems in various industrial and defence sectors is also increasing continuously. Since the operating threshold voltages and currents of these devices are very small they are very prone to electrical disturbance in their operation by the Electromagnetic Interference (EMI) signals. Their proper functioning is very important particularly in the case of systems used in mission mode, critical defence/industrial platforms. EMI can be generated within the electronic system/equipment itself or may result due to some external electromagnetic source. The high power Ultra Wide Band system is one such kind of external High Power Electromagnetic (HPEM) interference source which may cause malfunctioning/physical damage to the sensitive electronic systems. Hence it is necessary to test the susceptibility of electronics to such high power UWB based intentional EMI or IEMI sources. The sources for generating these transient EM fields may also be used in impulse radars and offensive applications to mal-operate/damage non-friendly electronics. The UWB system consists of a high voltage pulsed power source called pulser along with a high bandwidth (Ultra Wide Band) antenna to radiate the UWB signal. The pulse fed by the pulser to the antenna through a switch is of high voltage type (amplitude of few 10s of kV to about a MV) and has a sub-nanosecond rise time. Most of the UWB systems developed over the world have the switch employing gaseous dielectric switching media used at pressures above the atmospheric level to generate such a fast rise time voltage pulse. Use of gaseous switching media at sub-atmospheric pressures to achieve sub-ns rise time, short duration high voltage pulses required for the high power UWB applications is another possibility. This possibility has not been exploited till date. Hence it was decided to develop a pulser switch with gaseous switching media at sub-atmospheric pressures (up to 50 mbar) and achieve sub-ns rise time voltage pulses of up to 50 kV. The energy delivered out by the UWB system depends upon the pulser output energy per switching shot and the repetitive switching rate of the pulser. To achieve maximum energy output it is required to maximize either the energy per switching shot or the pulse repetition rate (PRR) of the pulser switch. The optimization of the pulser operation to achieve maximum pulser energy output in every switching shot has not been tried so far. In this work it was decided to analyze the circuit so as to achieve maximum pulser output energy per switching shot. Another objective of the study was to systematically characterize the pulser switch using various gases and gas mixtures as the switching media to evaluate the switch performance as a function of gas pressure and switch breakdown voltage. The effect of pulser and antenna performance parameters on the UWB system performance was also decided to be evaluated. Hence the present thesis work deals with the design, development, evaluation and performance optimization of a 50 kV, 25 MW UWB system based on Half Impulse Radiating Antenna (HIRA) fed by a coaxial capacitive pulser. The spark gap type self triggered pulser switch is designed to have a fixed gap spacing and variable gas pressure in order to vary the switch breakdown voltage. The switch is designed for operation with dry air, nitrogen, sulphur hexafluoride (SF6) and a mixture of different gases as the dielectric switching media with pressures of up to 5 bar above the atmospheric level and up to 50 mbar below the atmospheric level. Physical placement of the switch just above the coaxial pulser capacitor terminal offered a low inductance geometry. The rise time estimation of the switch has been carried out as a function of gas pressure and the switch arc inductance. These rise time values have been compared with the measured ones and a good agreement was found between the two. The rise time values indicate that an inverse relationship exists between the gas pressure and the rise time. The rise time was found to decrease at increased pressures. SF6 gas offered the minimum rise time out of all the gases/mixtures studied. The pulse repetition rate (PRR) of the UWB system depends upon the dielectric recovery of the gaseous switch and the charging time of the pulser capacitor. To estimate the PRR a circuit model has been proposed based on these parameters. The model shows an inverse relationship between the switch breakdown voltage (BDV) and the gas pressure with the PRR. The estimated PRR values were found to vary between 800 Hz and 5 kHz in the experimented range of the switch breakdown voltage. The PRR values have also been experimentally measured. There is a good match between the measured and the estimated values up to the switch BDV of 12.5 kV after which the difference is increased to about 20 %. The feed for the reflector of the HIRA antenna consists of a pair of coplanar conical transverse electromagnetic (TEM) feed plates as they have a better antenna aperture blockage performance. The angles of the TEM feed plates have been chosen using stereographic projections of the feed plates into the HIRA reflector. Each TEM feed plate of 200  characteristic impedance has been terminated by matched resistor. An analytical expression has been derived to optimize the pulser output voltage at which the energy output per switching shot of the UWB system is maximum. It was found that when the pulser output voltage i.e. the switch breakdown voltage is 75 % of the dc source voltage the output energy delivered is maximum. It was possible to achieve a maximum output energy of 10 J per switching shot for the designed 25 MW high power UWB system. The HIRA antenna has been analysed for the impedance profile for frequencies up to 3.5 GHz and was found to maintain a reflection performance better than -10 dB over the frequency range. The radiated field analysis of the antenna was carried out using an analytical model and numerically by using a commercially available software. It was found that as per the analytical model, the Figure of Merit (FoM) of the designed UWB system is 1.41 V for a normalized excitation feed pulse of 1 V and the 3 dB spectral content of the radiated field is between 180 MHz-1.8 GHz. The corresponding results using computer simulations of the UWB system indicate a slightly lesser FoM of 1.1. Higher FoM obtained using the analytical model is due to ignoring the antenna aperture blockage and the field diffraction effects over the TEM feed arms as well as from the rim of the reflector of the antenna. The radiated field amplitude and gain of the HIRA antenna were found to be a direct function of the frequency of the radiated signal. Higher gains and narrower beam width for the radiated field were observed with an increase in the frequency. The radiated field spectral waveform in the near field region was observed to have a notch at a particular frequency and its harmonics. The notch frequency was found to be a function of the propagation time difference called clear time. The effect of pulser rise time, antenna feed arm impedance and position on the radiated far field amplitude and wave shape was analysed. It was observed that with decrease in the pulser rise time from 700 ps to 100 ps, the radiated field amplitude increases by about 600 %. A matched termination impedance with position of 30of the TEM feed arms with respect to the vertical symmetry axis of the antenna provides a higher radiated field amplitude and lower post pulse oscillations in the radiated field waveform. The pulser switch was evaluated systematically for various performance parameters such as BDV, rise time, PRR, voltage recovery and jitter characteristics as a function of switch gas pressure, type of gaseous switching media and breakdown voltage at pressures above and below the atmospheric level. The switch BDV was found to be a linear function of pressure of the gas used i.e. dry air, nitrogen, sulphur hexafluoride (SF6) and a mixture of air and SF6. The measured rise times of all the gases were found to be in inverse proportion to the switch gas pressure. SF6 gas offered the best rise time and hence was found to be a good contender for achieving higher radiated field amplitudes and bandwidth. The voltage recovery characteristics of SF6 gas and air were experimentally studied as a function of the recovery time. It is found that both the gases have similar recovery characteristics having a distinct saturation plateau region. It was found that for a given recovery time SF6 recovers to a higher voltage than air and the recovery further improves for SF6 at increased pressures (between 0.5-2 bar). The effect of the number of switching shots on the jitter in the switch rise time was measured by operating the switch continuously at a PRR of 1 kHz and for total number shots up to 10.8 M. It was observed that the jitter increases by an order of magnitude after 10.8 M shots. This indicates that for the present switch design, the switch electrodes require maintenance (buffing, polishing, etc.) after every 3.5 M shots to maintain a reasonably low jitter. SF6 gas was characterized for a fixed source voltage to determine the effect of pressure on rise time in the sub atmospheric regime (up to 50 mbar). It was found that the rise time vs. pressure characteristics follows the Paschen’s curve with a value of pressure at which rise time is the lowest for a given source voltage. With increase in the source voltage the rise time was found to decrease. The HIRA based UWB radiating system was evaluated for radiated fields in the near and far field region for the temporal and spectral characteristics. It was found that for the source voltage of 25 kV, the FoM in the near and far field region are 29.4 kV and 28.9 kV respectively. The fields in the distant far field region have more oscillatory post pulses due to the effect of ground reflections and the low frequency dipole moment mismatch of the antenna. Since SF6 gas offered the best rise time of 193 ps at a voltage of 46 kV than the other gases tried, the radiated field is the highest (5.3 kV/m) with SF6 at a distance of 10 m offering a gain factor of 1.15. Dry air offered a radiated field gain factor of 0.83 which got improved by 33 % by just 30 % addition of SF6 gas into the air. The field amplitudes measured were in good agreement with those computed using the analytical model and the computer simulations and they follow the 1/R rule as a function of the far field distance, R in the bore sight direction. The measured radiation pattern of the UWB system showed a focussed and narrow radiated field beam at higher frequencies with a half field beam width (HFBW) of 8 at 2 GHz. The UWB system was measured to have dominant highest cut off frequency of 1.79 GHz with a band ratio and percentage band width of 9.56 and 162.11 % respectively. This confirmed that the developed system is of sub-hyper band radiator type. The UWB system developed through this work is having a better performance than some of the other systems developed elsewhere in the world, in terms of FoM (53 kV) and the PRR (> 1 kHz). The system can be further improved in terms of consistency (jitter) and intensity by use of a triggered switch and hydrogen gas at 100 bar pressure as the switching medium respectively. The profile of the TEM feed plates of the HIRA antenna may be further improved to have a better antenna aperture fill factor. Such multiple systems in an arrayed manner may be used either for higher power output/better agility of the radiated field beam. This system will be fully exploited for the applications of susceptibility evaluation of electronic circuits, non-friendly applications as well as impulse radars

The development of new high throughput technologies able to multiplex disease biomarkers as well as advances in medical treatments has lead to the recent expansion of the newborn screening panel to include DNA-based targets. Four rare disorders; deletion 22q11.2 syndrome and Spinal Muscular Atrophy (SMA), Severe Combined Immunodeficiency (SCID) and Congenital Cytomegalovirus (CMV), are potential candidates for inclusion to the newborn screening panel within the next few years. The major focus of this study was to determine whether 5’-hydrolysis assays developed for the four distinct disorders with specific detection needs and analytical ranges could be combined on the OpenArray system and in multiplexed qPCR reactions. SNP detection of homozygous SMN1 deletions in SMA, CNV detection in the 22q11.2 critical region, and quantification of the SCID biomarker, T-cell receptor excision circles (TRECs) and CMV were all required for disease confirmation. SMA and 22q11.2 gene deletions were accurately detected using the OpenArray system, a first for the technology. The medium density deletion 22q11.2 multiplex successfully identified deletion carriers having either the larger 3 Mb deletion or the smaller 1.5 Mb deletions. Both TREC and CMV targets were detected but with a decrease in sensitivity when compared to their singleplex counterparts. Lastly, copy number detection of the TBX1 was performed when multiplexed with the TREC assay, without a decrease in detection limit of either assay. Here, we provide proof of principal that qPCR multiplexing technologies are amenable to implementation with a newborn screening laboratory.

The present thesis is concerned with the genesis, diffusion, impact and institutionalisation of economic ideas. Despite Keynes's oft-cited comments to the effect that 'the ideas of economists and political philosophers, both when they are right and when they are wrong, are more powerful than is commonly understood'(Keynes 1936: 383), and the highly visible impact of economic ideas (for example Keynesian economics, Monetarism, or economic ideas regarding deregulation and antitrust issues) on the economic system, economists have done little to systematically explore the spread and impact of economic ideas. In fact, with only a few notable exceptions, the majority of scholarly work concerning the spread and impact of economic ideas has been developed outside of the economics literature, for example in the political institutionalist literature in the social sciences. The present thesis addresses the current lack of attention to the spread and impact of economic ideas by economists by drawing on the political institutionalist, sociological, and psychology of creativity literatures to develop a framework in which the genesis, spread, impact and institutionalisation of economic ideas may be understood. To articulate the dissemination and impact of economic ideas within economics, I consider as a case study the evolution of economists' conception of the economic agent - "homo oeconomicus." I argue that the intellectual milieu or paradigm of economics is 'socially constructed' in a specific sense, namely: (i) economic ideas are created or modified by particular individuals; (ii) economic ideas are disseminated (iii) certain economic ideas are accepted by economists and (iv) economic ideas become institutionalised into the paradigm or milieu of economics. Economic ideas are, of course, disseminated not only within economics to fellow economists, but are also disseminated externally to economic policy makers and business leaders who can - and often do - take economic ideas into account when formulating policy and building economic institutions. Important economic institutions are thereby socially constructed, in the general sense proposed by Berger and Luckmann (1966). But how exactly do economic ideas enter into this process of social construction of economic institutions? Drawing from and building on structure/agency theory (e.g. Berger and Luckmann 1966; Bourdieu 1977; Bhaskar 1979/1998, 1989; Bourdieu 1990; Lawson 1997, 2003) in the wider social sciences, I provide a framework for understanding how economic ideas enter into the process of social construction of economic institutions. Finally, I take up a methodological question: if economic ideas are disseminated, and if economic ideas have a real and constitutive impact on the economic system being modelled, does 'economic science' then accurately and objectively model an independently existing economic reality, unchanged by economic theory, or does economic theory have an interdependent and 'reflexive' relationship with economic reality, as economic reality co-exists with, is shaped by, and also shapes economic theory? I argue the latter, and consider the implications for evaluating in what sense economic science is, in fact, a science in the classical sense. The thesis makes original contributions to understanding the genesis of economic ideas in the psychological creative work processes of economists; understanding the ontological location of economic ideas in the economic system; articulating the social construction of economic ideas; and highlighting the importance of the spread of economic ideas to economic practice and economic methodology.