Chez les bactéries, la régulation de l’expression génétique est fondamentale pour permettre une adaptation optimale à l’environnement. De nombreux contrôles existent, notamment au niveau post-transcriptionnel par de nombreux ARN régulateurs (sRNA pour « small RNAs »). Ceux-ci ciblent des ARN messager (ARNm), permettant une régulation rapide de la synthèse de protéines. Le plus souvent, ces sRNAs interagissent avec leur(s) cible(s) au niveau du site de fixation du ribosome (RBS pour « ribosome binding site »), entrant dès lors en compétition avec le ribosome pour la fixation à l’ARNm et entraînant une régulation négative de l’expression des gènes cibles. Pour autant, il existe de nombreux mécanismes alternatifs de régulation par les sRNA. Nous avons ainsi pu démontrer que les deux sRNAs OmrA et OmrB, conservés au sein des entérobactéries, répriment la synthèse du récepteur FepA aux complexes fer-entérobactine en ciblant une structure de l’ARNm fepA. Cette structure en tige-boucle est située en aval du RBS de fepA, et de façon surprenante, elle contrôle positivement la synthèse de FepA via une activation de la fixation de la sous-unité 30S du ribosome à l’ARNm. Des structures similaires ont pu être prédites dans d’autres ARNm, à l’image de bamA, codant la sous-unité essentielle du complexe Bam d’adressage des protéines de membrane externe en tonneaux β. Comme pour fepA, la tige-boucle de l'ARNm bamA favorise la fixation du ribosome, suggérant que ce mécanisme de régulation pourrait être bien plus général du fait de la grande conservation de bamA au sein des bactéries à Gram négatif. De surcroît, ces résultats constituent la première illustration que les structures d'ARNm peuvent avoir un effet positif sur la traduction. Par ailleurs, deux autres sRNAs répriment également et indépendamment l’expression de fepA, à savoir SdsR et RseX. A chaque fois, le mécanisme de régulation impliqué est différent. Ainsi, SdsR s’apparie vraisemblablement à deux régions différentes de l’ARNm fepA, impliquant notamment une compétition classique avec la fixation du ribosome. La répression par RseX nécessite quant à elle la présence d’autres séquences du 5’UTR de fepA, à plus d’une centaine de nucléotides (nts) en amont du RBS. Enfin, chacun de ces sRNAs semble répondre à des stimuli différents, ce qui enrichit considérablement notre connaissance des signaux contribuant à la régulation de fepA, dont jusqu’ici seule la carence en fer était connue comme un signal de dérépression par le facteur de transcription Fur. Ce travail est une nouvelle illustration de l’immense diversité des mécanismes de régulation impliquant des ARN, dont la grande flexibilité de structure et de séquence constitue une importante source de diversité à la disposition de l’évolution / In order to perfectly adapt to their environment, bacteria require a tight gene expression regulation. This can occur through post-transcriptional control by numerous regulatory RNAs (or small RNAs, sRNAs). These sRNAs can target mRNAs, leading to a fast regulation of protein synthesis. Most often, sRNAs base-pair with their target mRNAs at the ribosome binding site (RBS), therefore competing with the ribosome for the binding with the mRNA and repressing gene expression. However, many other regulatory mechanisms involve sRNAs. We have demonstrated that the two sRNAs OmrA and OmrB, conserved among enterobacteria, repress the synthesis of the FepA receptor for iron-enterobactin complexes through base-pairing with a secondary structure within fepA mRNA. This stem-loop structure is located downstream of fepA RBS, and most surprisingly, promotes 30S ribosomal subunit binding to fepA mRNA, therefore activating FepA synthesis. Similar stem-loop structures have been predicted in other mRNAs, such as the bamA mRNA encoding the essential subunit of the Bam outer membrane protein assembly complex. As for fepA mRNA, the stem-loop found in bamA mRNA also promotes ribosome binding, showing that this regulatory mechanism could be widespread considering the strong conservation of bamA among Gram negative bacteria. Moreover, these results challenge the commonly admitted view of mRNA secondary structures being repressors of gene expression. Two other sRNAs also repress fepA expression in an OmrA/B-independent fashion, namely SdsR and RseX. For each of these sRNAs, the regulatory mechanism involved is different. Indeed, SdsR most likely acts through two distinct binding sites, one of which leading to a classical competition with the ribosome binding. Meanwhile, RseX repression requires most of fepA 5’UTR, including sequences at about 100nt upstream of the start codon. Finally, each of these sRNAs is expressed upon diverse stimuli, considerably extending our knowledge of the signals leading to fepA regulation, for which only the Fur-dependent derepression upon iron starvation was known. This work highlights the great versatility of regulatory mechanisms involving RNAs. This illustrates how RNAs structural flexibility and sequence diversity is a key source of diversity for evolution

Three folded maps (1 col.) in pocket. Bibliography: leaves 81-94.

Bibliography: p.127-141.

Typescript (Photocopy) Copies of 4 published papers co-authored by author, and 7 maps, in back cover pocket. Bibliography: leaves 203-218.

Stimuli responsive polymers (SRP) have attracted a lot of attention, due to their potential and promising applications in many fields, as protein-ligand recognition, on-off switches for modulated drug delivery or artificial organs. Poly(N-isopropylacrylamide) (PNIPAM) is one of the most widely studied polymers due to its lower critical solution temperature (LCST) at ~ 32° C in aqueous solution. Additionally, glycopolymers, where free sugar units are present, have potentially interesting applications especially in bio-recognition where sugars play an important role. In this work, our interest was focused on the synthesis of glycomonomers and its block- and random- copolymers with NIPAM. NIPAM homopolymers with an active chain transfer unit at the chain end could be prepared by RAFT. They were used as macro-chain transfer agents to prepare a variety of sugar containing responsive block copolymers from new glycomonomers by the monomer addition concept. The LCSTs of the aqueous solutions of the copolymers are affected strongly by the comonomer content, spacer chain length of the glycomonomer and the chain architecture of the copolymers. These polymers were coated on a solid substrate by spin coating and crosslinked by plasma immobilization. Characterization of the polymers was performed by nuclear magnetic resonance spectroscopy (NMR), ultraviolet (UV), dynamic light scattering (DLS, detection of aggregation behaviour) and gel permeation chromatography (GPC). Polymer films were investigated by ellipsometry, X-ray photoelectron spectroscopy (XPS) and atomic force microscopy (AFM) regarding their surface properties. Afterwards sulfation of sugar – OH groups was performed in order to obtain heparin like structure, as heparin exhibits numerous important biological activities, like good interaction with diverse proteins. Finally, affinity of the polymers (sulfated and non sulfated form) on a solid support to the endothelial cells was investigated.

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Η Τομογραφία Εκπομπής Ποζιτρονίων, η οποία συχνά αναφέρεται με βάση το ακρωνύμιό της, PET (Positron Emission Tomography), αποτελεί μία πρωτοποριακή τεχνική απεικόνισης η οποία παρέχει εγκάρσιες τομές της λειτουργίας των διαφόρων δομών του ανθρωπίνου σώματος. Η Τομογραφία PET επιτρέπει την μεταβολική απεικόνιση αυτών των δομών (σε αντίθεση με τις ακτίνες-Χ και την Υπολογιστική Τομογραφία (CT – Computer Tomography) οι οποίες παρέχουν ανατομική απεικόνιση), σε μοριακό επίπεδο, και αυτός είναι ο λόγος που συχνά η Τομογραφία PET αναφέρεται ως μοριακή απεικόνιση. Ειδικότερα, οι τομογράφοι PET για μικρά ζώα (Small Animal PET) οι οποίοι απαιτούν ιδιαίτερα υψηλή διακριτική ικανότητα, διαδραματίζουν σημαντικό ρόλο στην βιολογία και στις in-vivo μελέτες της φαρμακοκινητικής των ιχνηθετών και του μεταβολισμού. Σύμφωνα με τη βιβλιογραφία έχουν αναφερθεί διάφοροι τύποι τομογράφων PET, στους οποίους χρησιμοποιήθηκαν διάφοροι τύποι ανιχνευτικών διατάξεων, με διαφορετικό σχεδιασμό σε κάθε περίπτωση. Η τεχνολογία αυτών των τομογράφων βασίζεται στη χρήση μικρών ανόργανων κρυστάλλων, κυρίς αποτελούμενων από BGO, GSO και LSO, οι οποίοι σχηματίζουν ένα block στο οποίο έχει προσαρτηθεί ένας φωτοπολλαπλασιαστής είτε με ευαισθησία θέσης (PS – PMT: Position Sensitive PhotoMultiplier Tube), είτε με πολλαπλές ανόδους (multianode PMT). To BGO (Bismuth Germanate Oxide) είναι το υλικό που χρησιμοποιείται σε αρκετούς εμπορικούς Τομογράφους, έχοντας πλέον αντικαταστήσει το ιωδιούχο νάτριο (ΝαΙ). Ένα πρότυπο σύστημα small animal PET χαμηλού κόστους βρίσκεται υπό ανάπτυξη, προκειμένου να μελετήσουμε τα επιμέρους σχεδιαστικά χαρακτηριστικά του και να μετρήσουμε την απόδοσή του. Ο βασικός block ανιχνευτής αποτελείται από μία 16×16 διάταξη επιμέρους BGO κρυστάλλων διαστάσεων 3.75×3.75×20 mm3, ο οποίος έχει τοποθετηθεί με ειδική διεργασία στην επιφάνεια ενός Hamamatsu R-2486 PSPMT. Με τη χρήση κατάλληλων ηλεκτρονικών διατάξεων και την ανάπτυξη ειδικού λογισμικού πραγματοποιήθηκαν μετρήσεις της απόδοσης των επιμέρους ανιχνευτών καθώς επίσης και μετρήσεις σχετικές με τους φωτοπολλαπλασιαστές. / Positron Emission Tomography, often referred to by its acronym, PET, is an emerging radiologic modality that yields transverse tomographic images of functioning organs in the human body. PET enables the metabolic imaging of organs (as opposed to the anatomic imaging provided by techniques such as the X-ray imaging or the Computerized Tomography (CT)), in molecular level, and this is the reason why it is characterized as molecular imaging. Especially, small animal PET tomographs which require high spatial resolution can play an important role in biology and studies of in vivo tracer pharmacokinetics and metabolism. Various implementations have been reported in the literature using a variety of detector and design technologies. The basic technology for these scanners is based on small inorganic crystals, mainly from BGO, GSO, and LSO, forming detector blocks read out by position sensitive and multianode PMTs. BGO is the material used in a lot of commercial scanners, having replaced NaI, mainly because BGO has higher stopping power and it is not hygroscopic. We have been developing a low-cost small animal PET prototype, in order to study specific design characteristics and measure its performance. The basic block detector design consists of a 16×16 array of 3.75×3.75×20 mm3 individual BGO crystals coupled to a Hamamatsu R-2486 PSPMT. Measurements of the individual detector performance as well as measurements of the PSPMTs have been performed.

We have effectively identified and removed ground roll through a twostep process. The first step is to identify the major components of the ground roll through various methods including multiscale separation, directional or frequency filtering or by any other method that identifies the ground roll. Given this estimate for ground roll, the recorded signal is separated during the second step through a block-coordinate relaxation method that seeks the sparsest set for weighted curvelet coefficients of the ground roll and the sought-after reflectivity. The combination of these two methods allows us to separate out the ground roll signal while preserving the reflector information. Since our method is iterative, we have control of the separation process. We successfully tested our algorithm on a real data set with a complex ground roll and reflector structure.

Age-related inhibitory effects were investigated during spatial memory performance. In Experiment 1, 15 young (M = 20 years) and 16 old adults (M = 70 years) completed two spatial tasks (i.e., Block Suppression Test, Corsi Block Tapping Test) that differed in need for inhibitory processing. Accuracy differences within each task revealed age-related differences in spatial working memory and between task differences revealed that older adults had difficulty ignoring irrelevant items. Experiment 2 (10 young, 10 old adults) examined whether the distractibility of irrelevant items in the inhibition task (i.e. BST) accounted for the age-related inhibitory effects. Findings were largely consistent with the initial experiment indicating that inhibitory function was affected by adult aging.

Bibliography: p.127-141. / xv, 210 p. : ill. (some col.), maps (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Geology, 2001?