ARTD1/PARP1 Negatively Regulates Glycolysis by Inhibiting Hexokinase 1 Independent of NAD(+) Depletion

ARTD1 (PARP1) is a key enzyme involved in DNA repair through the synthesis of poly(ADP-ribose) (PAR) in response to strand breaks, and it plays an important role in cell death following excessive DNA damage. ARTD1-induced cell death is associated with NAD(+) depletion and ATP loss; however, the molecular mechanism of ARTD1-mediated energy collapse remains elusive. Using real-time metabolic measurements, we compared the effects of ARTD1 activation and direct NAD(+) depletion. We found that ARTD1-mediated PAR synthesis, but not direct NAD(+) depletion, resulted in a block to glycolysis and ATP loss. We then established a proteomics-based PAR interactome after DNA damage and identified hexokinase 1 (HK1) as a PAR binding protein. HK1 activity is suppressed following nuclear ARTD1 activation and binding by PAR. These findings help explain how prolonged activation of ARTD1 triggers energy collapse and cell death, revealing insight into the importance of nucleus-to-mitochondria communication via ARTD1 activation.

Shared Pseudomonas aeruginosa genotypes are common in Australian cystic fibrosis centres

Recent molecular-typing studies suggest cross-infection as one of the potential acquisition pathways for Pseudomonas aeruginosa in patients with cystic fibrosis (CF). In Australia, there is only limited evidence of unrelated patients sharing indistinguishable P. aeruginosa strains. We therefore examined the point-prevalence, distribution, diversity and clinical impact of P. aeruginosa strains in Australian CF patients nationally. 983 patients attending 18 Australian CF centres provided 2887 sputum P. aeruginosa isolates for genotyping by enterobacterial repetitive intergenic consensus-PCR assays with confirmation by multilocus sequence typing. Demographic and clinical details were recorded for each participant. Overall, 610 (62%) patients harboured at least one of 38 shared genotypes. Most shared strains were in small patient clusters from a limited number of centres. However, the two predominant genotypes, AUST-01 and AUST-02, were widely dispersed, being detected in 220 (22%) and 173 (18%) patients attending 17 and 16 centres, respectively. AUST-01 was associated with significantly greater treatment requirements than unique P. aeruginosa strains. Multiple clusters of shared P. aeruginosa strains are common in Australian CF centres. At least one of the predominant and widespread genotypes is associated with increased healthcare utilisation. Longitudinal studies are now needed to determine the infection control implications of these findings.

Multi-centre research in Australia:analysis of a recent National Health and Medical Research Council-funded project

BACKGROUND AND OBJECTIVE: Human research ethics committees provide essential review of research projects to ensure the ethical conduct of human research. Several recent reports have highlighted a complex process for successful application for human research ethics committee approval, particularly for multi-centre studies. Limited resources are available for the execution of human clinical research in Australia and around the world.

METHODS: This report overviews the process of ethics approval for a National Health and Medical Research Council-funded multi-centre study in Australia, focussing on the time and resource implications of such applications in 2007 and 2008.

RESULTS: Applications were submitted to 16 hospital and two university human research ethics committees. The total time to gain final approval from each committee ranged between 13 and 77 days (median = 46 days); the entire process took 16 months to complete and the research officer's time was estimated to cost $A34 143.

CONCLUSIONS: Obstacles to timely human research ethics committee approval are reviewed, including recent, planned and potential initiatives that could improve the ethics approval of multi-centre research.

Low rates of Pseudomonas aeruginosa misidentification in isolates from cystic fibrosis patients

Pseudomonas aeruginosa is an important cause of pulmonary infection in cystic fibrosis (CF). Its correct identification ensures effective patient management and infection control strategies. However, little is known about how often CF sputum isolates are falsely identified as P. aeruginosa. We used P. aeruginosa-specific duplex real-time PCR assays to determine if 2,267 P. aeruginosa sputum isolates from 561 CF patients were correctly identified by 17 Australian clinical microbiology laboratories. Misidentified isolates underwent further phenotypic tests, amplified rRNA gene restriction analysis, and partial 16S rRNA gene sequence analysis. Participating laboratories were surveyed on how they identified P. aeruginosa from CF sputum. Overall, 2,214 (97.7%) isolates from 531 (94.7%) CF patients were correctly identified as P. aeruginosa. Further testing with the API 20NE kit correctly identified only 34 (59%) of the misidentified isolates. Twelve (40%) patients had previously grown the misidentified species in their sputum. Achromobacter xylosoxidans (n = 21), Stenotrophomonas maltophilia (n = 15), and Inquilinus limosus (n = 4) were the species most commonly misidentified as P. aeruginosa. Overall, there were very low rates of P. aeruginosa misidentification among isolates from a broad cross section of Australian CF patients. Additional improvements are possible by undertaking a culture history review, noting colonial morphology, and performing stringent oxidase, DNase, and colistin susceptibility testing for all presumptive P. aeruginosa isolates. Isolates exhibiting atypical phenotypic features should be evaluated further by additional phenotypic or genotypic identification techniques.

Granular columns for stabilizing slopes

A series of small-scale tests was undertaken to verify if granular anchors could be used as a slope stabilisation technique. The nature of the material used and the resulting loading configuration are described here. The work confirms that the inclusion of anchors within a slope mass, irrespective of their number or orientation, significantly enhances the capacity and ductility of the failure mode. The small-scale nature of this research did influence the observed capacities, but the overarching hypothesis was confirmed. A simple analysis method is proposed that allows designers to accurately remediate natural or man-made slopes using existing analytical methods for slope stability.

Strengthening timber beams with near surface mounted carbon fiber reinforced polymer rods

Many timber structures may require strengthening due to either decay and aging or an increase of load. This paper presents an experimental study in which eleven timber beams were tested, including three unstrengthened reference beams and eight beams strengthened with NSM CFRP bars. The test parameters include the position of NSM (tensile face or the bottom of the sides), the number of CFRP bars (1 or 2), and additional anchorage of NSM CFRP bars (steel wire U anchors or CFRP U strips). The test results show that the ultimate flexural strength of the timber beams were increased by 14%∼85% with an average of 47% due to NSM CFRP bar strengthening. Their deflection corresponding to the peak load was increased by 33% in average.

O binómio jornalista-editor de imagem na produção noticiosa televisiva: causas e consequências

Na era da convergência jornalística, as redações televisivas estão, ainda, a passar por diversos processos de transformações. Centrando esta problemáti-ca nos telejornalistas, verifica-se uma tendência crescente para que estes de-sempenhem funções e/ou tarefas que ainda há muito pouco tempo eram exe-cutadas por outros profissionais da informação televisiva. Esta agregação fun-cional verifica-se, nomeadamente, ao nível da edição de imagem de conteúdos noticiosos a serem transmitidos pela televisão. Neste contexto, na presente tese de doutoramento pretende-se, em termos gerais, apurar e compreender as causas e as consequências de serem os próprios jornalistas televisivos a desempenhar a edição de imagem de conteúdos noticiosos transmitidos pela televisão. Para isso, foi desenvolvida uma investigação empírica, tendo como campos de estudo as redações centrais da SIC e da TVI. Tal como o trabalho de campo permitiu concluir, os diversos fundamen-tos desta dinâmica são, de certa forma, incontornáveis. Contudo, esta investi-gação pretende também contribuir para que se reúnam as condições, ao nível da oferta formativa, que tornem as respetivas consequências consonantes com uma evolução profícua da atividade jornalística.

Decentralizing IP mobility management in future networks

The massive adoption of sophisticated mobile devices and applications led to the increase of mobile data in the last decade, which it is expected to continue. This increase of mobile data negatively impacts the network planning and dimension, since core networks are heavy centralized. Mobile operators are investigating atten network architectures that distribute the responsibility of providing connectivity and mobility, in order to improve the network scalability and performance. Moreover, service providers are moving the content servers closer to the user, in order to ensure high availability and performance of content delivery. Besides the e orts to overcome the explosion of mobile data, current mobility management models are heavy centralized to ensure reachability and session continuity to the users connected to the network. Nowadays, deployed architectures have a small number of centralized mobility anchors managing the mobile data and the mobility context of millions of users, which introduces issues related to performance and scalability that require costly network mechanisms. The mobility management needs to be rethought out-of-the box to cope with atten network architectures and distributed content servers closer to the user, which is the purpose of the work developed in this Thesis. The Thesis starts with a characterization of mobility management into well-de ned functional blocks, their interaction and potential grouping. The decentralized mobility management is studied through analytical models and simulations, in which di erent mobility approaches distinctly distribute the mobility management functionalities through the network. The outcome of this study showed that decentralized mobility management brings advantages. Hence, it was proposed a novel distributed and dynamic mobility management approach, which is exhaustively evaluated through analytical models, simulations and testbed experiments. The proposed approach is also integrated with seamless horizontal handover mechanisms, as well as evaluated in vehicular environments. The mobility mechanisms are also speci ed for multihomed scenarios, in order to provide data o oading with IP mobility from cellular to other access networks. In the pursuing of the optimized mobile routing path, a novel network-based strategy for localized mobility is addressed, in which a replication binding system is deployed in the mobility anchors distributed through the access routers and gateways. Finally, we go further in the mobility anchoring subject, presenting a context-aware adaptive IP mobility anchoring model that dynamically assigns the mobility anchors that provide the optimized routing path to a session, based on the user and network context. The integration of dynamic and distributed concepts in the mobility management, such as context-aware adaptive mobility anchoring and dynamic mobility support, allow the optimization of network resources and the improvement of user experience. The overall outcome demonstrates that decentralized mobility management is a promising direction, hence, its ideas should be taken into account by mobile operators in the deployment of future networks.

Structural analysis of glycans and development of microarrays from Helcobacter pylori cell surface glycome

Helicobacter pylori is a bacterial pathogen that affects more than half of the world’s population with gastro-intestinal diseases and is associated with gastric cancer. The cell surface of H. pylori is decorated with lipopolysaccharides (LPSs) composed of three distinct regions: a variable polysaccharide moiety (O-chain), a structurally conserved core oligosaccharide, and a lipid A region that anchors the LPS to the cell membrane. The O-chain of H. pylori LPS, exhibits unique oligosaccharide structures, such as Lewis (Le) antigens, similar to those present in the gastric mucosa and are involved in interactions with the host. Glucan, heptoglycan, and riban domains are present in the outer core region of some H. pylori LPSs. Amylose-like glycans and mannans are also constituents of some H. pylori strains, possibly co-expressed with LPSs. The complexity of H. pylori LPSs has hampered the establishment of accurate structure-function relationships in interactions with the host, and the design of carbohydrate-based therapeutics, such as vaccines. Carbohydrate microarrays are recent powerful and sensitive tools for studying carbohydrate antigens and, since their emergence, are providing insights into the function of carbohydrates and their involvement in pathogen-host interactions. The major goals of this thesis were the structural analysis of LPSs from H. pylori strains isolated from gastric biopsies of symptomatic Portuguese patients and the construction of a novel pathogen carbohydrate microarray of these LPSs (H. pylori LPS microarray) for interaction studies with proteins. LPSs were extracted from the cell surface of five H. pylori clinical isolates and one NCTC strain (26695) by phenol/water method, fractionated by size exclusion chromatography and analysed by gas chromatography coupled to mass spectrometry. The oligosaccharides released after mild acid treatment of the LPS were analysed by electrospray mass spectrometry. In addition to the conserved core oligosaccharide moieties, structural analyses revealed the presence of type-2 Lex and Ley antigens and N-acetyllactosamine (LacNAc) sequences, typically found in H. pylori strains. Also, the presence of O-6 linked glucose residues, particularly in LPSs from strains 2191 and NCTC 26695, pointed out to the expression of a 6-glucan. Other structural domains, namely ribans, composed of O-2 linked ribofuranose residues were observed in the LPS of most of H. pylori clinical isolates. For the LPS from strain 14382, large amounts of O-3 linked galactose units, pointing to the occurrence of a galactan, a domain recently identified in the LPS of another H. pylori strain. A particular feature to the LPSs from strains 2191 and CI-117 was the detection of large amounts of O-4 linked N-acetylglucosamine (GlcNAc) residues, suggesting the presence of chitin-like glycans, which to our knowledge have not been described for H. pylori strains. For the construction of the H. pylori LPS microarray, the structurally analysed LPSs, as well as LPS-derived oligosaccharide fractions, prepared as neoglycolipid (NGL) probes were noncovalently immobilized onto nitrocellulosecoated glass slides. These were printed together with NGLs of selected sequence defined oligosaccharides, bacterial LPSs and polysaccharides. The H. pylori LPS microarray was probed for recognition with carbohydratebinding proteins (CBPs) of known specificity. These included Le and blood group-related monoclonal antibodies (mAbs), plant lectins, a carbohydratebinding module (CBM) and the mammalian immune receptors DC-SIGN and Dectin-1. The analysis of these CBPs provided new information that complemented the structural analyses and was valuable in the quality control of the constructed microarray. Microarray analysis revealed the occurrence of type-2 Lex and Ley, but not type-1 Lea or Leb antigens, supporting the results obtained in the structural analysis. Furthermore, the H. pylori LPSs were recognised by DC-SIGN, a mammalian lectin known to interact with this bacterium through fucosylated Le epitopes expressed in its LPSs. The -fucose-specific lectin UEA-I, showed restricted binding to probes containing type-2 blood group H sequence and to the LPSs from strains CI-117 and 14382. The presence of H-type-2, as well Htype- 1 in the LPSs from these strains, was confirmed using specific mAbs. Although H-type-1 determinant has been reported for H. pylori LPSs, this is the first report of the presence of H-type-2 determinant. Microarray analysis also revealed that plant lectins known to bind 4-linked GlcNAc chitin oligosaccharide sequences bound H. pylori LPSs. STL, which exhibited restricted and strong binding to 4GlcNAc tri- and pentasaccharides, differentially recognised the LPS from the strain CI-117. The chitin sequences recognised in the LPS could be internal, as no binding was detected to this LPS with WGA, known to be specific for nonreducing terminal of 4GlcNAc sequence. Analyses of the H. pylori LPSs by SDS-PAGE and Western blot with STL provided further evidence for the presence of these novel domains in the O-chain region of this LPS. H. pylori LPS microarray was also applied to analysis of two human sera. The first was from a case infected with H. pylori (H. pylori+ CI-5) and the second was from a non-infected control.The analysis revealed a higher IgG-reactivity towards H. pylori LPSs in the H. pylori+ serum, than the control serum. A specific IgG response was observed to the LPS isolated from the CI-5 strain, which caused the infection. The present thesis has contributed to extension of current knowledge on chemical structures of LPS from H. pylori clinical isolates. Furthermore, the H. pylori LPS microarray constructed enabled the study of interactions with host proteins and showed promise as a tool in serological studies of H. pyloriinfected individuals. Thus, it is anticipated that the use of these complementary approaches may contribute to a better understanding of the molecular complexity of the LPSs and their role in pathogenesis.

La clarté cognitive et l'entrée dans l'écrit

Partant de la diversité des compétences des enfants face à l'écrit en entrant en 1re année, nous avons vérifié, si dès la maternelle, une intervention portant directement sur la représentation de la langue écrite, de la clarté cognitive et de la conscience linguistique pouvait favoriser les premiers apprentissages formels (lecture et écriture). L'échantillon auprès duquel a été menée l'expérimentation était constituée de 4 groupes d'enfants de maternelle (5 ans) provenant de milieu socio-économique moyen; 2 de ces groupes ont reçu une stimulation à l'écrit tandis que les autres ont suivi le programme du MEQ. Notre plan d'expérimentation partant des théories de Downing, Fijalkow et Ferreiro s'est déroulé sur une période de neuf mois. L'analyse des résultats nous permet d'avancer que l'intervention sur l'écrit à la maternelle favorise l'accroissement de la clarté cognitive et contribue à élargir leur conscience linguistique.

Les pratiques de gouvernance de commissions scolaires du Québec

Le système d'éducation public québécois actuel est marqué par la complexité de son fonctionnement multijoueurs amplifiée par un accroissement des prescriptions législatives en matière de gouvernance et de procédures participatives. Devant ce constat, l'objectif de cette thèse est de décrire et de comprendre, à partir de l'analyse de pratiques d'acteurs, le fonctionnement réel de la gouvernance de deux commissions scolaires (CS) québécoises en regard de ce qui est prescrit par la Loi. Utilisant une approche qualitative, notre recherche repose sur une étude de cas dans laquelle trois dossiers sont étudiés pour chacune des organisations. L' analyse est menée à partir des concepts propres aux approches théoriques de l'analyse stratégique et du néoinstitutionnalisme. Nous concluons de cette étude de pratiques de gouvernance que les dossiers étudiés sont menés conformément aux prescriptions législatives, voire que ces pratiques vont souvent au-delà des exigences législatives, notamment quant aux procédures de participation. Dans les CS étudiés, les processus de gouvernance mis en oeuvre s'appuient généralement sur un porteur de dossier et des comités de travail regroupant des représentants-relais de plusieurs acteurs concernés. Ce fonctionnement est observé au sein des deux organisations, et ce, même si les modes de coopération présentent des différences notables. Au niveau des acteurs, dans les deux cas étudiés, les cadres de la CS et le Conseil des commissaires sont perçus comme étant très influents sur le déroulement des dossiers. L'influence des autres acteurs, tels que les parents, les représentants de la communauté et les acteurs associés aux établissements, est perçue comme étant variable selon les organisations et selon les dossiers. Les perceptions quant à la marge de manoeuvre et aux capacités d'influence des acteurs, l'intérêt porté au dossier et la personnalité des individus impliqués s'avèrent des facteurs marquants quant à cette inégalité.

Étude CFD des effets du désalignement et du cisaillement sur les performances et le chargement des hydroliennes

Le développement des hydroliennes se base souvent sur des conditions d'écoulement idéalisées qui ne reflètent pas entièrement le courant présent sur un site réel. Dans cette optique, ce mémoire investigue l'effet d'un écoulement non aligné ou cisaillé sur les performances et le chargement de deux types d'hydrolienne : les hydroliennes à rotor axial (HRA) et à aile oscillante (HAO). Cette étude est réalisée à l'aide de simulations numériques et les résultats démontrent que pour les deux types d'hydrolienne, le désalignement produit une réduction de la puissance et de la traînée, tandis que le cisaillement n'a qu'un effet de second ordre sur ces derniers. Le chargement additionnel sur la structure de support causé par des conditions d'écoulement non idéal est aussi similaire pour les deux technologies. Le désalignement et le cisaillement affectent cependant plus significativement le chargement en fatigue des pales de l'HRA que celui de l'aile de l'HAO.

Sexe de l’enfant et investissement parental: le cas d’Haïti

Bon nombre d’études révèlent que, dans les pays en développement, la préférence des parents pour un sexe en particulier les pousse souvent à traiter leurs enfants inéquitablement. Pour déterminer si ce phénomène existe en Haïti, cette étude s’inspire des travaux de Barcellos et al. (2014) sur les parents indiens dont les résultats, obtenus à partir des données d’enquête démographique et de santé (DHS 1992), suggèrent une discrimination au détriment des filles. Cette discrimination a lieu au niveau du temps et des ressources consacrés à prendre soin de l’enfant suite à sa naissance. Les résultats obtenus à l’aide des données nationales révèlent qu’en 1994, parmi les variables d’intérêt (allaitement, immunisation, vitamine A), seule la probabilité d’allaitement semble varier par sexe en faveur des garçons en Haïti. Tout nouveau-né de sexe masculin a une probabilité de 3,2 points de pourcentage plus élevée d’être allaité que le sexe opposé. Des données plus récentes révèlent que le sexe de l’enfant n’a aucun impact sur la probabilité d’être allaité en 2000. Par contre, en 2005 l’effet de cette probabilité est contraire à celui observé en 1994. En l’occurrence, si le nouveau-né est de sexe féminin elle a 2,4 points de pourcentage de plus que les garçons d’être allaité. De plus, considérant les enfants de 0 à 59 mois, les estimations révèlent qu’en 2005 la durée de l’allaitement augmente de 11,2% si l’enfant est une fille. Mots clés: Investissement, Haïti, enfant, sexe, allaitement, mère, immunisation, anthropométrie, 1994, 2000, 2005.

Jazz Combos, March 5, 2003

Concert program for Jazz Combos, March 5, 2003

Hydrogen peroxide sensing, signaling and regulation of transcription factors

The regulatory mechanisms by which hydrogen peroxide (H2O2) modulates the activity of transcription factors in bacteria (OxyR and PerR), lower eukaryotes (Yap1, Maf1, Hsf1 and Msn2/4) and mammalian cells (AP-1, NRF2, CREB, HSF1, HIF-1, TP53, NF-κB, NOTCH, SP1 and SCREB-1) are reviewed. The complexity of regulatory networks increases throughout the phylogenetic tree, reaching a high level of complexity in mammalians. Multiple H2O2 sensors and pathways are triggered converging in the regulation of transcription factors at several levels: (1) synthesis of the transcription factor by upregulating transcription or increasing both mRNA stability and translation; (ii) stability of the transcription factor by decreasing its association with the ubiquitin E3 ligase complex or by inhibiting this complex; (iii) cytoplasm-nuclear traffic by exposing/masking nuclear localization signals, or by releasing the transcription factor from partners or from membrane anchors; and, (iv) DNA binding and nuclear transactivation by modulating transcription factor affinity towards DNA, co-activators or repressors, and by targeting specific regions of chromatin to activate individual genes. We also discuss how H2O2 biological specificity results from diverse thiol protein sensors, with different reactivity of their sulfhydryl groups towards H2O2, being activated by different concentrations and times of exposure to H2O2. The specific regulation of local H2O2 concentrations is also crucial and results from H2O2 localized production and removal controlled by signals. Finally, we formulate equations to extract from typical experiments quantitative data concerning H2O2 reactivity with sensor molecules. Rate constants of 140 M-1s−1 and ≥ 1.3 × 103 M-1s−1 were estimated, respectively, for the reaction of H2O2 with KEAP1 and with an unknown target that mediates NRF2 protein synthesis. In conclusion, the multitude of H2O2 targets and mechanisms provides an opportunity for highly specific effects on gene regulation that depend on the cell type and on signals received from the cellular microenvironment.