Measuring surface potential changes on leaves.

We provide here a detailed protocol for studying the changes in electrical surface potential of leaves. This method has been developed over the years by plant physiologists and is currently used in different variants in many laboratories. The protocol records surface potential changes to measure long-distance electrical signals induced by diverse stimuli such as leaf wounding or current injection. This technique can be used to determine signaling speeds, to measure the connectivity between different plant organs and-by exploiting mutant plants-to identify transporters and ion channels involved in electrical signaling. The approach can be combined with the analysis of mRNA expression and of metabolite concentrations to correlate electrical signaling to specific physiological events. We describe how to use this protocol on Arabidopsis, looking at the effects of leaf wounding; however, it is broadly applicable to other plants and can be used to study other aspects of plant physiology. After wound infliction, surface potential recording takes ∼20 min per plant.

The structural dynamics of US output and inflation: What explains the changes?

We examine the dynamics of US output and inflation using a structural time varyingcoefficient VAR. We show that there are changes in the volatility of both variables andin the persistence of inflation. Technology shocks explain changes in output volatility,while a combination of technology, demand and monetary shocks explain variations inthe persistence and volatility of inflation. We detect changes over time in the transmission of technology shocks and in the variance of technology and of monetary policyshocks. Hours and labor productivity always increase in response to technology shocks.

Grey matter changes in motor conversion disorder.

OBJECTIVE: To detect anatomical differences in areas related to motor processing between patients with motor conversion disorder (CD) and controls. METHODS: T1-weighted 3T brain MRI data of 15 patients suffering from motor CD (nine with hemiparesis and six with paraparesis) and 25 age- and gender-matched healthy volunteers were compared using voxel-based morphometry (VBM) and voxel-based cortical thickness (VBCT) analysis. RESULTS: We report significant cortical thickness (VBCT) increases in the bilateral premotor cortex of hemiparetic patients relative to controls and a trend towards increased grey matter volume (VBM) in the same region. Regression analyses showed a non-significant positive correlation between cortical thickness changes and symptom severity as well as illness duration in CD patients. CONCLUSIONS: Cortical thickness increases in premotor cortical areas of patients with hemiparetic CD provide evidence for altered brain structure in a condition with presumed normal brain anatomy. These may either represent premorbid vulnerability or a plasticity phenomenon related to the disease with the trends towards correlations with clinical variables supporting the latter.

Do institutional changes affect business cycles? Evidence from Europe

We study the effects that the Maastricht treaty, the creation of the ECB, andthe Euro changeover had on the dynamics of European business cycles using a panelVAR and data from ten European countries - seven from the Euro area and threeoutside of it. There are changes in the features of European business cycles and in thetransmission of shocks. They precede the three events of interest and are more linkedto a general process of European convergence and synchronization.

Processes and consequences in business ethical dilemmas: The oil industry and climate changes

This paper proposes a framework to examine business ethical dilemmas andbusiness attitudes towards such dilemmas. Business ethical dilemmas canbe understood as reflecting a contradiction between a socially detrimentalprocess and a self-interested profitable consequence. This representationallows us to distinguish two forms of behavior differing by whetherpriority is put on consequences or on processes. We argue that theseforms imply very different business attitudes towards society:controversial or competitive for the former and aligned or cooperativefor the latter. These attitudes are then analyzed at the discursive level in order to address the question of good faith in businessargumentation, i.e. to which extent are these attitudes consistent withactual business behaviors. We argue that consequential attitudes mostlyinvolve communication and lobbying actions aiming at eluding the dilemma.Therefore, the question of good faith for consequential attitudes liesin the consistency between beliefs and discourse. On the other hand,procedural attitudes acknowledge the dilemma and claim a change of theprocess of behavior. They thus raise the question of the consistencybetween discourses and actual behavior. We apply this processes/consequencesframework to the case of the oil industry s climate change ethical dilemmawhich comes forth as a dilemma between emitting greenhouse gases and making more profits . And we examine the different attitudes of two oilcorporations-BP Amoco and ExxonMobil-towards the dilemma.

Retinal degeneration progression changes lentiviral vector cell targeting in the retina

Ce travail de thèse a été réalisé au sein de l'Unité de Thérapie Génique et Biologie des Cellules Souches de l'Hôpital Jules- Gonin dans le Service d'Ophtalmologie de l'Université de Lausanne. Ce laboratoire recherche des solutions thérapeutiques pour des maladies dégénératives et incurables de la rétine comme les rétinites pigmentaires (RP). Ayant déjà montré certains résultats dans le domaine, la thérapie génique a été notre outil pour ce travail. Cette méthode se base sur le principe de remplacer un gène déficient par sa copie normale, en transportant celle-ci au coeur même du noyau par un vecteur. Il existe à l'heure actuelle différents vecteurs. Un des plus efficaces est un vecteur viral non-réplicatif : le lentivirus, dérivé de HIV-1. Celui-ci a la capacité d'intégrer le génome de la cellule cible, lui conférant ainsi un nouveau matériel génétique. Notre but a été d'établir le tropisme du lentivirus dans une rétine en dégénérescence. Ce lentivirus est connu pour transduire efficacement les cellules de l'épithélium pigmentaire rétinien dans l'oeil adulte sain, ainsi que celles de la neurorétine, mais ce, uniquement durant le développement. On sait aussi que le vecteur lentiviral présente un tropisme différent selon les enveloppes dont il est muni ; par exemple, le lentivirus avec une enveloppe Mokola est connu pour transduire les cellules gliales du système nerveux central. La rétine qui dégénère montre quant à elle des changements de sa structure qui pourraient influencer la diffusion du vecteur et/ou son tropisme. Le postulat de base a été le suivant : chez l'adulte, la transduction des neurones de la rétine via le lentivirus pourrait être facilitée par l'altération de la membrane limitante externe induite par la dégénérescence (meilleure pénétrance du virus). D'un point de vue technique, nous avons utilisé deux types distincts de modèles murins de dégénérescence rétinienne : des souris Balb/C soumises à une dose toxique de lumière et les souris Rhodopsin knockout, animaux génétiquement modifiés. Comme vecteur viral, nous avons employé deux différents pseudotypes de lentivirus (caractérisés par les enveloppes virales) avec différents promoteurs (séquence d'ADN qui initie la transduction et confère la spécificité d'expression d'un gène). En changeant l'enveloppe et le promoteur, nous avons essayé de trouver la meilleure combinaison pour augmenter l'affinité du vecteur vis-à-vis des photorécepteurs d'abord, puis vis-à-vis d'autres cellules de la rétine. Nos résultats ont montré que la membrane limitante externe est effectivement altérée chez les deux modèles de dégénérescence, mais que cette modification ne favorise pas la transduction des photorécepteurs lorsqu'on utilise un vecteur lentiviral contenant une enveloppe VSVG et un promoteur photorécepteur-spécifique ou ubiquitaire. En effet, une forte réaction gliale a été observée. Par contre, en utilisant le lentivirus avec une enveloppe Mokola et un promoteur ubiquitaire, nous avons constaté une très bonne transduction au niveau des cellules de Millier dans la rétine en dégénérescence, phénomène non observé chez les souris sauvages. Ce travail a donc permis de trouver un vecteur viral efficace pour atteindre et transduire les cellules de Miiller, ceci seulement pendant la dégénérescence de la rétine. Ces cellules, une fois transduites, pourraient être utilisées pour sécréter dans la rétine des agents thérapeutiques tels que des facteurs neurotrophiques pour soutenir la survie des photorécepteurs ou des facteurs anti-angiogéniques pour prévenir la néo-vascularisation lors de diabète ou de dégénérescence maculaire liée à l'âge. - In normal mice, the lentiviral vector (LV) is very efficient to target the RPE cells, but transduces retinal neurons well only during development. In the present study, the tropism of LV has been investigated in the degenerating retina of mice, knowing that the retina structure changes during degeneration. We postulated that the viral transduction would be increased by the alteration of the iuter limiting membrane (OLM). Two different LV pseudotypes were tested using the VSVG arid the Mokola envelopes, as well as two animal models of retinal degeneration: light-damaged Balb-C and Rhodopsin knockout (Rho-/-) mice. After light damage, the OLM is altered and no significant increase of the number of transduced photoreceptors can be obtained with a LV-VSVG-Rhop-GFP vector. In the Rho-/- mice, an altération of the OLM was also observed, but the possibility of transducing photoreceptors was decreased, probably by ongoing gliosis. The use of a ubiquitous promoter allows better photoreceptor transduction, suggesting that photoreceptór-specific promoter activity change during late stages of photoreceptor degeneration. However, the number of targeted photoreceptors remains low. In contrast, LV pseudotyped with the tfokola envelope allows a wide dispersion of the ctor into the retina (corresponding to the injection bleb) with preferential targeting of Muller cells, a situation Mc\ does ot occur in the wild- type retina. Mokola-pseudotyped lentiviral vectors may serve to engineer these glial cells to deliver secreted therapeutic factors to a diseased area of the retina.

Personality traits, cognition and volumetric MRI changes in elderly patients with early-onset depression: A 2-year follow-up study.

Previous studies revealed personality changes in elderly patients with early-onset depression (EOD) that persist in euthymic stages. However, depression in older patients is a complex disorder that may affect not only personality, but also cognition and brain structure. To address this issue, a cross-sectional comparison and 2-year follow-up of 28 EOD elderly patients and 48 healthy controls included detailed neurocognitive assessment, estimates of brain volumes in limbic areas and white matter hyperintensities, as well as evaluation of the Five Factor Model of personality, in a remitted mood state. Results revealed that cognitive performances as well as brain volumes were preserved in EOD patients both at baseline and at follow-up. The increased Neuroticism factor and Anxiety facet scores as well as the decreased Warmth and Positive Emotions facet scores found at baseline reached the level of healthy controls after 2years. Only the Depression facet scores remained significantly higher in EOD patients compared to controls upon follow-up. Results were independent of depressive relapse since baseline (25% of patients). These findings suggest that both cognitive performances and brain volumes show long-term preservation in older EOD patients. In contrast, the depression-related personality facet might be a trait like marker that persists in the long-term evolution of this disorder.

Assessment of microbial community structure changes by amplified ribosomal DNA restriction analysis (ARDRA)

Amplified ribosomal DNA restriction analysis (ARDRA) is a simple method based on restriction endonuclease digestion of the amplified bacterial 16S rDNA. In this study we have evaluated the suitability of this method to detect differences in activated sludge bacterial communities fed on domestic or industrial wastewater, and subject to different operational conditions. The ability of ARDRA to detect these differences has been tested in modified Ludzack-Ettinger (MLE) configurations. Samples from three activated sludge wastewater treatment plants (WWTPs) with the MLE configuration were collected for both oxic and anoxic reactors, and ARDRA patterns using double enzyme digestions AluI+MspI were obtained. A matrix of Dice similarity coefficients was calculated and used to compare these restriction patterns. Differences in the community structure due to influent characteristics and temperature could be observed, but not between the oxic and anoxic reactors of each of the three MLE configurations. Other possible applications of ARDRA for detecting and monitoring changes in activated sludge systems are also discussed

Transcript profiling suggests that differential metabolic adaptation of mice to a high fat diet is associated with changes in liver to muscle lipid fluxes.

Genetically homogenous C57Bl/6 mice display differential metabolic adaptation when fed a high fat diet for 9 months. Most become obese and diabetic, but a significant fraction remains lean and diabetic or lean and non-diabetic. Here, we performed microarray analysis of "metabolic" transcripts expressed in liver and hindlimb muscles to evaluate: (i) whether expressed transcript patterns could indicate changes in metabolic pathways associated with the different phenotypes, (ii) how these changes differed from the early metabolic adaptation to short term high fat feeding, and (iii) whether gene classifiers could be established that were characteristic of each metabolic phenotype. Our data indicate that obesity/diabetes was associated with preserved hepatic lipogenic gene expression and increased plasma levels of very low density lipoprotein and, in muscle, with an increase in lipoprotein lipase gene expression. This suggests increased muscle fatty acid uptake, which may favor insulin resistance. In contrast, the lean mice showed a strong reduction in the expression of hepatic lipogenic genes, in particular of Scd-1, a gene linked to sensitivity to diet-induced obesity; the lean and non-diabetic mice presented an additional increased expression of eNos in liver. After 1 week of high fat feeding the liver gene expression pattern was distinct from that seen at 9 months in any of the three mouse groups, thus indicating progressive establishment of the different phenotypes. Strikingly, development of the obese phenotype involved re-expression of Scd-1 and other lipogenic genes. Finally, gene classifiers could be established that were characteristic of each metabolic phenotype. Together, these data suggest that epigenetic mechanisms influence gene expression patterns and metabolic fates.

Changes of beta-amyloid precursor protein splice patterns in brain cell aggregate cultures.

The splice pattern of beta-amyloid precursor protein (beta-APP) has been studied in a variety of neuronal and glial cells and in brain cell aggregate cultures by the polymerase chain reaction (PCR). The brain-typical pattern, in which beta-APP695 is the dominant form, has been found only in aggregate cultures but not in any of the other cell types including neuronal cell lines. Selective elimination of glial cells from aggregates resulted in increased quantities of beta-APP695, whereas removal of neurons led to a reduction of beta-APP695 and to an elevation of beta-APP751 and beta-APP770. This shift of splice pattern was not observed in cocultures of the neuronal cell line PC 12 with primary astrocytes combined in a variety of cellular ratios. Blood serum, which is an essential component of these cultures, tested on aggregates, did not reduce the amount of beta-APP695 or have any marked effects on splice patterns generally. From these results it is concluded that investigations on brain-typical splicing of beta-APP require primary neurons. Neuronal cell lines may be no suitable model systems. Splicing events favoring production of beta-APP695 may mark an important, very early step of amyloid formation in the brain.

Changes in understorey biomass and species composition after water level drawdown on pine mires in southern Finland

Tiivistelmä: Vedenpinnan alenemisen vaikutus sararämeen pintakasvillisuuden biomassaan ja lajistoon

Chemical changes in argisols under irrigated grape production in the central São Francisco river valley, Brazil

This study compares the chemical composition of the solution and exchange complex of soil in a 3-year-old irrigated vineyard (Vitis vinifera L., Red Globe cultivar) with that of adjacent clearing in the native hyperxerophyllic 'caatinga' vegetation. The soils are classified as Plinthic Eutrophic Red-Yellow Argisol; according to Soil Taxonomy they are isohyperthermic Plinthustalfs. Detailed physiographic characterization revealed an impermeable gravel and cobble covering the crystalline rocks; the relief of this layer was more undulating than the level surface. Significant higher concentrations of extractable Na, K, Mg and Ca were observed within the vineyard. Lower soil acidity, higher Ca/Mg ratios, as well as lower sodium adsorption and Na/K ratios reflected additions of dolomitic lime, superphosphate and K-bearing fertilizers. As the water of the São Francisco River is of good quality for irrigation (C1S1), the increases in Na were primarily attributed to capillary rise from the saline groundwater table. None of the soil in the study area was found to be sodic. About 62% of the vineyard had an Ap horizon with salinity levels above 1.5dSm-1 (considered detrimental for grape production); according to average values for this horizon, a potential 13% reduction in grape production was predicted. Differences in chemical composition in function of distance to the collector canals were observed in the clearing, but not in the vineyard. The influence of differences in the elevations of the surface and impermeable layers, as well as pediment thickness, was generally weaker under irrigation. Under irrigation, soil moisture was greater in points of convergent surface waterflow; the effect of surface curvature on chemical properties, though less consistent, was also stronger in the vineyard.

A phylogenetic study of the phosphoenolpyruvate carboxylase multigene family in Poaceae: understanding the molecular changes linked to C4 photosynthesis evolution

Phylogenetic reconstructions have supported several independent appearances of C₄ photosynthesis within grasses (Poaceae). These recurrent appearances appear to contradict the large number of biochemical and morphological changes required to change from C₃ to C₄, a paradox that leads to questions about the genetic changes underlying C₄ evolution. In this study, we analysed sequences encoding phosphoenolpyruvate carboxylases (PEPCs) in grasses in order to gain insights into the origin of the ppc-C₄ gene, which encodes a key enzyme in the C₄ pathway. We screened databanks for PEPC genes or cDNAs in grasses. A coding sequence of 1130 base pairs was used to build phylogenetic trees that supported the existence of four distinct PEPC gene lineages. Ppc-C₄ present in all C₄ grasses was also found in two C₃ species. The ppc-C₄ clade was congruent with the species tree, suggesting orthologous evolution. This result would imply that ppc-C₄ appeared without any duplication event. Nevertheless, caution is needed since the sampling of our study is still far from comprehensive. Further investigation with an increased sampling is recommended to elucidate the evolutionary changes underlying ppc-C₄ gene evolution in grasses.