Long-distance wound signalling in arabidopsis

The leaves of all plants use elaborate and inducible defence systems to protect themselves. A wide variety of such defences are known and they include defence chemicals such as alkaloids, phenolics and terpenes, physical structures ranging from fibre cells to silica deposits, and a wide variety of defence proteins many of which target digestive processes in herbivores. It has long been known that the defence responses of plants under attack by insects are not restricted to the site of attack. Instead, if a leaf is damaged, defence can be triggered in other parts of the plant body, for example in distal leaves or even in roots and flowers. This raises the question of what are the organ-to-organ signals that coordinate this process. Several hypotheses have been proposed. These include the long-distance transfer of chemical signals through the plant vasculature, hydraulic signals that may transit through the xylem, and electrical signals that would move through living tissues such as the phloem. Much evidence for each of these scenarios has been published. In this thesis we took advantage of the fact that many plant defence responses are regulated by a signal transduction pathway based on a molecule called jasmonic acid. We used this molecule, one of its derivatives (jasmonoyl-isoleucine), and some of the genes it regulates as markers. Using these we investigated the possible role of the electrical signals in the leaf- to-leaf activation of the jasmonate pathway. We found that feeding insects stimulate easily detected electrical activity in the leaves of Arabidopsis thaliana and we used non-invasive surface electrodes to record this activity. This approach showed that jasmonate pathway activity and the electrical activity provoked by mechanical wounding occurred within identical spatial boundaries. Measurements of the apparent speed of surface potentials agreed well with previous velocity estimates for the speed of leaf-to-leaf signals that activate the jasmonate pathway. Using this knowledge we were able to investigate the effects of current injection into Arabidopsis leaves. This resulted in the strong expression of many jasmonate-regulated genes. All these results showed that electrical activity and the activation of jasmonate signalling were highly correlated. In order to test for possible causal links between the two processes, we conducted a small-scale reverse genetic screen on a series of T-DNA insertion mutants in ion channel genes and in other genes encoding proteins such as proton pumps. This screen, which was based on surface potential measurements, revealed that mutations in genes related to ionotropic glutamate receptors in animals had impaired electrical activity after wounding. Combining mutation of two of these glutamate-receptor-like genes in a double mutant reduced the response of leaves to current injection. When a leaf of this double mutant was wounded it failed to transmit a long-distance signal to a distal leaf. This result distinguished the double mutant from the wild-type plant and provides the first genetic evidence that electrical signalling is necessary to coordinate defence responses between organs in plants. - Les feuilles des plantes disposent de systèmes de défense inductibles très élaborés. Un grand nombre de ces systèmes de défenses sont connus et sont basés sur des composés chimiques comme les alcaloïdes, les composés phénoliques ou les terpènes, des systèmes physiques allant de la production de cellules fibreuses aux cristaux de silice ainsi qu'un grand nombre de protéines de défense ciblant le processus digestif des herbivores. Il est connu dépuis longtemps que la réponse défensive de la plante face à l'attaque pas un insecte n'est pas seulement localisée au niveau de la zone d'attaque. A la place, si une feuille est attaquée, les systèmes de défense peuvent être activés ailleurs dans la plante, comme par exemple dans d'autres feuilles, les racines ou même les fleurs. Ces observations soulèvent la question de la nature des signaux d'organes à organes qui régulent ces systèmes. Plusieurs hypothèses ont été formulées; une ou plusieures molécules pourraient être véhiculées dans la plante grâce au système vasculaire, un signal hydraulique transmis au travers du xylème ou encore des signaux électriques transmis par les cellules comme dans le phloème par exemple. De nombreuses études ont été publiées sur ces différentes hypothèses. Dans ce travail de thèse, nous avons choisi d'utiliser à notre avantage le fait que de nombreuses réponses de défense de la plante sont régulées par une même voie de signalisation utilisant l'acide jasmonique. Nous avons utilisé comme marqueurs cette molécule, un de ses dérivés (le jasmonoyl-isoleucine) ainsi que certains des gènes que l'acide jasmonique régule. Nous avons alors testé l'implication de la transmission de signaux électriques dans l'activation de la voie du jasmonate de feuille à feuille. Nous avons découvert que les insectes qui se nourrissent de feuilles d'Arabidopsis thaliana activent un signal électrique que nous avons pu mesurer grâce à une technique non invasive d'électrodes de surface. Les enregistrements ont montré que la génération de signaux électriques et l'activation de la voie du jasmonate avaient lieu aux mêmes endroits. La mesure de la vitesse de déplacement des impulsions électriques correspond aux estimations faites concernant l'activation de la voie du jasmonate. Grâce à cela, nous avons pu tester l'effet d'injection de courant électrique dans les feuilles d'Arabidopsis. La conséquence a été une forte expression de nombreux gènes de la voie du jasmonate, suggérant une forte corrélation entre l'activité électrique et l'activation de la voie du jasmonate. Afin de tester le lien de cause entre ces deux phénomènes, nous avons entrepris un criblage génétique sur une série de mutants d'insertion à l'ADN-T dans des gènes de canaux ioniques et d'autres gènes d'intérêt comme les gènes des pompes à protons. Ce criblage, basé sur la mesure de potentiels de surface, a permis de montrer que plusieurs mutations de gènes liés aux récepteurs au glutamate ionotropique présentent une baisse drastique de leurs activités électriques après une blessure mécanique des feuilles par rapport au type sauvage. Par la combinaison de deux mutations de ces récepteurs au glutamate en un double mutant, on obtient une réponse à la stimulation électrique encore plus faible. Quand une feuille du double mutant est blessée, elle est incapable de transmettre un signal à longue distance vers une feuille éloignée. Ce résultat permet de distinguer le double mutant de la plante sauvage et amène la première preuve génétique que l'activité électrique est nécessaire pour coordonner les réponses de défense entre les organes chez les plantes.

Body fluid and tissue analysis using filter paper sampling support prior to LC-MS/MS: application to fatal overdose with colchicine

Because of the various matrices available for forensic investigations, the development of versatile analytical approaches allowing the simultaneous determination of drugs is challenging. The aim of this work was to assess a liquid chromatography-tandem mass spectrometry (LC-MS/MS) platform allowing the rapid quantification of colchicine in body fluids and tissues collected in the context of a fatal overdose. For this purpose, filter paper was used as a sampling support and was associated with an automated 96-well plate extraction performed by the LC autosampler itself. The developed method features a 7-min total run time including automated filter paper extraction (2 min) and chromatographic separation (5 min). The sample preparation was reduced to a minimum regardless of the matrix analyzed. This platform was fully validated for dried blood spots (DBS) in the toxic concentration range of colchicine. The DBS calibration curve was applied successfully to quantification in all other matrices (body fluids and tissues) except for bile, where an excessive matrix effect was found. The distribution of colchicine for a fatal overdose case was reported as follows: peripheral blood, 29 ng/ml; urine, 94 ng/ml; vitreous humour and cerebrospinal fluid, < 5 ng/ml; pericardial fluid, 14 ng/ml; brain, < 5 pg/mg; heart, 121 pg/mg; kidney, 245 pg/mg; and liver, 143 pg/mg. Although filter paper is usually employed for DBS, we report here the extension of this alternative sampling support to the analysis of other body fluids and tissues. The developed platform represents a rapid and versatile approach for drug determination in multiple forensic media.

Wipe sampling procedure coupled to LC-MS/MS analysis for the simultaneous determination of 10 cytotoxic drugs on different surfaces.

A simple wipe sampling procedure was developed for the surface contamination determination of ten cytotoxic drugs: cytarabine, gemcitabine, methotrexate, etoposide phosphate, cyclophosphamide, ifosfamide, irinotecan, doxorubicin, epirubicin and vincristine. Wiping was performed using Whatman filter paper on different surfaces such as stainless steel, polypropylene, polystyrol, glass, latex gloves, computer mouse and coated paperboard. Wiping and desorption procedures were investigated: The same solution containing 20% acetonitrile and 0.1% formic acid in water gave the best results. After ultrasonic desorption and then centrifugation, samples were analysed by a validated liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) in selected reaction monitoring mode. The whole analytical strategy from wipe sampling to LC-MS/MS analysis was evaluated to determine quantitative performance. The lowest limit of quantification of 10 ng per wiping sample (i.e. 0.1 ng cm(-2)) was determined for the ten investigated cytotoxic drugs. Relative standard deviation for intermediate precision was always inferior to 20%. As recovery was dependent on the tested surface for each drug, a correction factor was determined and applied for real samples. The method was then successfully applied at the cytotoxic production unit of the Geneva University Hospitals pharmacy.

Irregular to regular sampling, denoising and deconvolution

We propose a restoration algorithm for band limited images that considers irregular(perturbed) sampling, denoising, and deconvolution. We explore the application of a family ofregularizers that allow to control the spectral behavior of the solution combined with the irregular toregular sampling algorithms proposed by H.G. Feichtinger, K. Gr¨ochenig, M. Rauth and T. Strohmer.Moreover, the constraints given by the image acquisition model are incorporated as a set of localconstraints. And the analysis of such constraints leads to an early stopping rule meant to improvethe speed of the algorithm. Finally we present experiments focused on the restoration of satellite images, where the micro-vibrations are responsible of the type of distortions we are considering here. We will compare results of the proposed method with previous methods and show an extension tozoom.

Oxygen uptake kinetics and middle distance swimming performance.

OBJECTIVE: The aim of this study was to determine whether V˙O(2) kinetics and specifically, the time constant of transitions from rest to heavy (τ(p)H) and severe (τ(p)S) exercise intensities, are related to middle distance swimming performance. DESIGN: Fourteen highly trained male swimmers (mean ± SD: 20.5 ± 3.0 yr; 75.4 ± 12.4 kg; 1.80 ± 0.07 m) performed an discontinuous incremental test, as well as square wave transitions for heavy and severe swimming intensities, to determine V˙O(2) kinetics parameters using two exponential functions. METHODS: All the tests involved front-crawl swimming with breath-by-breath analysis using the Aquatrainer swimming snorkel. Endurance performance was recorded as the time taken to complete a 400 m freestyle swim within an official competition (T400), one month from the date of the other tests. RESULTS: T400 (Mean ± SD) (251.4 ± 12.4 s) was significantly correlated with τ(p)H (15.8 ± 4.8s; r=0.62; p=0.02) and τ(p)S (15.8 ± 4.7s; r=0.61; p=0.02). The best single predictor of 400 m freestyle time, out of the variables that were assessed, was the velocity at V˙O(2max)vV˙O(2max), which accounted for 80% of the variation in performance between swimmers. However, τ(p)H and V˙O(2max) were also found to influence the prediction of T400 when they were included in a regression model that involved respiratory parameters only. CONCLUSIONS: Faster kinetics during the primary phase of the V˙O(2) response is associated with better performance during middle-distance swimming. However, vV˙O(2max) appears to be a better predictor of T400.

How geographic distance and depth drive ecological variability and isolation of demersal fish communities in an archipelago system (Cape Verde, Eastern Atlantic Ocean)

Cape Verde is a tropical oceanic ecosystem, highly fragmented and dispersed, with islands physically isolated by distance and depth. To understand how isolation affects the ecological variability in this archipelago, we conducted a research project on the community structure of the 18 commercially most important demersal fishes. An index of ecological distance based on species relative dominance (Di) is developed from Catch Per Unit Effort, derived from an extensive database of artisanal fisheries. Two ecological measures of distance between islands are calculated: at the species level, DDi, and at the community level, DD (sum of DDi). A physical isolation factor (Idb) combining distance (d) and bathymetry (b) is proposed. Covariance analysis shows that isolation factor is positively correlated with both DDi and DD, suggesting that Idb can be considered as an ecological isolation factor. The effect of Idb varies with season and species. This effect is stronger in summer (May to November), than in winter (December to April), which appears to be more unstable. Species react differently to Idb, independently of season. A principal component analysis on the monthly (DDi) for the 12 islands and the 18 species, complemented by an agglomerative hierarchical clustering, shows a geographic pattern of island organization, according to Idb. Results indicate that the ecological structure of demersal fish communities of Cape Verde archipelago, both in time and space, can be explained by a geographic isolation factor. The analytical approach used here is promising and could be tested in other archipelago systems.

Distance education in neonatal nursing scenarios: a systematic review

OBJECTIVE Identify resources that support learning mediated by technology in the field of neonatal nursing. METHOD Systematic review with searches conducted in MEDLINE, LILACS and SciELO. Titles and abstracts were independently evaluated by two experts. RESULTS Of the 2,051 references, 203 full-text articles were analyzed, resulting in the inclusion of nine studies on semiotics and semiology, cardiopulmonary resuscitation, general aspects of neonatal care, diagnostic reasoning and assessment of pain. Only two articles addressed the development of educational strategies and seven papers described the assessment of these strategies by experts and/or users. CONCLUSION Distance education is an important resource for education, and its improvement and updating, and it particularly adds advantages for neonatal nursing by approximating teaching and real-life situations and by minimizing the exposure of newborns for teaching purposes. The lack of educational initiatives mediated by technology suggests the need for the development, evaluation and dissemination of educational resources focused on nursing care of newborns and their families.

Consumer Advisory Bulletin, Hang up on long distance slamming!, January 2008

The Attorney General’s Consumer Protection Division receives hundreds of calls and consumer complaints every year. Follow these tips to avoid unexpected expense and disappointments. This record is about: Hang up on long distance slamming!

Mise en Place d’un Dispositif de Formation a Distance a l’universite du Cap-Vert« e-learning » :Les Technologies de L’information et de la Communication dans L’enseignement du Français Langue Etrangere

Ce mémoire est l’occasion de partager le résultat de la mise en place de ce dispositif de formation à distance que nous avons mené dans l’université du Cap-Vert. Dans la première partie, nous décrirons la nature de la politique éducative au Cap-Vert. Nous contextualiserons les principes de l’installation de l’université publique dans ce pays, ainsi que les intentions d’innovation pédagogique de cette université. La deuxième partie portera un regard complémentaire sur l’utilisation des TIC et de l’internet dans l’enseignement/apprentissage d’une langue, cas du français langue étrangère, et nous nous inspirerons des théories constructiviste et socio-constructiviste. Finalement, la troisième partie détaillera toutes les étapes de la conception et de la mise en place du dispositif de formation à distance. Dans cette troisième partie, nous aborderons dans un premier temps la question des enjeux et des risques du e-Learning et nous présenterons notre mission dans le projet « e-Learning.cv » mené par l’université. Puis, dans un deuxième temps nous analyserons quelques cours que nous avons mis en ligne, en sachant qu’un cours en ligne n’est pas la simple reproduction d’un support pédagogique imprimé mais il offre à l’apprenant un environnement multimédia et interactif. Finalement dans un troisième temps nous essayerons de prendre un peu de recul pour faire une analyse critique de ce que nous avons réalisé et essayer par là même de dégager les perspectives pour améliorer le travail effectué.

Aplicação dos Sistemas de Informação Geográfica em Estudos de Caracterização Biofísica da Ilha de Santo Antão em Cabo Verde

Com características morfológicas e edafo-climáticas extremamente diversificadas, a ilha de Santo Antão em Cabo Verde apresenta uma reconhecida vulnerabilidade ambiental a par de uma elevada carência de estudos científicos que incidam sobre essa realidade e sirvam de base à uma compreensão integrada dos fenómenos. A cartografia digital e as tecnologias de informação geográfica vêm proporcionando um avanço tecnológico na colecção, armazenamento e processamento de dados espaciais. Várias ferramentas actualmente disponíveis permitem modelar uma multiplicidade de factores, localizar e quantificar os fenómenos bem como e definir os níveis de contribuição de diferentes factores no resultado final. No presente estudo, desenvolvido no âmbito do curso de pós-graduação e mestrado em sistemas de Informação geográfica realizado pela Universidade de Trás-os-Montes e Alto Douro, pretende-se contribuir para a minimização do deficit de informação relativa às características biofísicas da citada ilha, recorrendo-se à aplicação de tecnologias de informação geográfica e detecção remota, associadas à análise estatística multivariada. Nesse âmbito, foram produzidas e analisadas cartas temáticas e desenvolvido um modelo de análise integrada de dados. Com efeito, a multiplicidade de variáveis espaciais produzidas, de entre elas 29 variáveis com variação contínua passíveis de influenciar as características biofísicas da região e, possíveis ocorrências de efeitos mútuos antagónicos ou sinergéticos, condicionam uma relativa complexidade à interpretação a partir dos dados originais. Visando contornar este problema, recorre-se a uma rede de amostragem sistemática, totalizando 921 pontos ou repetições, para extrair os dados correspondentes às 29 variáveis nos pontos de amostragem e, subsequente desenvolvimento de técnicas de análise estatística multivariada, nomeadamente a análise em componentes principais. A aplicação destas técnicas permitiu simplificar e interpretar as variáreis originais, normalizando-as e resumindo a informação contida na diversidade de variáveis originais, correlacionadas entre si, num conjunto de variáveis ortogonais (não correlacionadas), e com níveis de importância decrescente, as componentes principais. Fixou-se como meta a concentração de 75% da variância dos dados originais explicadas pelas primeiras 3 componentes principais e, desenvolveu-se um processo interactivo em diferentes etapas, eliminando sucessivamente as variáveis menos representativas. Na última etapa do processo as 3 primeiras CP resultaram em 74,54% da variância dos dados originais explicadas mas, que vieram a demonstrar na fase posterior, serem insuficientes para retratar a realidade. Optou-se pela inclusão da 4ª CP (CP4), com a qual 84% da referida variância era explicada e, representando oito variáveis biofísicas: a altitude, a densidade hidrográfica, a densidade de fracturação geológica, a precipitação, o índice de vegetação, a temperatura, os recursos hídricos e a distância à rede hidrográfica. A subsequente interpolação da 1ª componente principal (CP1) e, das principais variáveis associadas as componentes CP2, CP3 e CP4 como variáveis auxiliares, recorrendo a técnicas geoestatística em ambiente ArcGIS permitiu a obtenção de uma carta representando 84% da variação das características biofísicas no território. A análise em clusters validada pelo teste “t de Student” permitiu reclassificar o território em 6 unidades biofísicas homogéneas. Conclui-se que, as tecnologias de informação geográfica actualmente disponíveis a par de facilitar análises interactivas e flexíveis, possibilitando que se faça variar temas e critérios, integrar novas informações e introduzir melhorias em modelos construídos com bases em informações disponíveis num determinado contexto, associadas a técnicas de análise estatística multivariada, possibilitam, com base em critérios científicos, desenvolver a análise integrada de múltiplas variáveis biofísicas cuja correlação entre si, torna complexa a compreensão integrada dos fenómenos.

When Preference Is Not Satisfied but the Individual Is: How Power Distance Affects Person-Job Fit

One aspect of person-job fit reflects congruence between personal preferences and job design; as congruence increases so should satisfaction. We hypothesized that power distance would moderate whether fit is related to satisfaction with degree of job formalization. We obtained measures of job-formalization, fit and satisfaction, as well as organizational commitment from employees (n = 772) in a multinational firm with subsidiaries in six countries. Confirming previous findings, individuals from low power-distance cultures were most satisfied with increasing fit. However, the extent to which individuals from high power-distance cultures were satisfied did not necessarily depend on increasing fit, but mostly on whether the degree of formalization received was congruent to cultural norms. Irrespective of culture, satisfaction with formalization predicted a broad measure of organizational commitment. Apart from our novel extension of fit theory, we show how moderation can be tested in the context of polynomial response surface regression and how specific hypotheses can be tested regarding different points on the response surface.

What risks do people perceive in everyday life? A perspective gained from the experience sampling method (ESM)

The experiential sampling method (ESM) was used to collect data from 74 parttimestudents who described and assessed the risks involved in their current activitieswhen interrupted at random moments by text messages. The major categories ofperceived risk were short-term in nature and involved loss of time or materials relatedto work and physical damage (e.g., from transportation). Using techniques of multilevelanalysis, we demonstrate effects of gender, emotional state, and types of risk onassessments of risk. Specifically, females do not differ from males in assessing thepotential severity of risks but they see these as more likely to occur. Also, participantsassessed risks to be lower when in more positive self-reported emotional states. Wefurther demonstrate the potential of ESM by showing that risk assessments associatedwith current actions exceed those made retrospectively. We conclude by notingadvantages and disadvantages of ESM for collecting data about risk perceptions.

Minimum distance estimation of stationary and non-stationary ARFIMA processes

A new parametric minimum distance time-domain estimator for ARFIMA processes is introduced in this paper. The proposed estimator minimizes the sum of squared correlations of residuals obtained after filtering a series through ARFIMA parameters. The estimator iseasy to compute and is consistent and asymptotically normally distributed for fractionallyintegrated (FI) processes with an integration order d strictly greater than -0.75. Therefore, it can be applied to both stationary and non-stationary processes. Deterministic components are also allowed in the DGP. Furthermore, as a by-product, the estimation procedure provides an immediate check on the adequacy of the specified model. This is so because the criterion function, when evaluated at the estimated values, coincides with the Box-Pierce goodness of fit statistic. Empirical applications and Monte-Carlo simulations supporting the analytical results and showing the good performance of the estimator in finite samples are also provided.