Expression of the proto-oncogene c-fos in three-dimensional fetal brain cell cultures and the lack of correlation with maturation-inducing stimuli.

Previous work has shown that aggregating fetal brain cell cultures are able to attain a highly differentiated state, and that their development is greatly enhanced by growth and/or differentiation factors such as epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and the protein kinase C-activating tumor promoter mezerein. The present study shows that in these 3-dimensional cultures the peptide growth factors EGF and bFGF as well as mezerein are able to induce the expression of the proto-oncogene c-fos. This induction was rapid and transient, in good agreement with observations reported from a wide variety of cell types in vitro. The maximal levels of c-fos mRNA found after stimulation were low in immature cultures and increased greatly as maturation progressed. Of the three factors tested, mezerein was the most potent inducer of c-fos. In contrast to the peptide growth factors EGF and bFGF which were found to induce c-fos only in glial cells, mezerein was stimulatory in glial cells as well as in neurons. A similar cell type specificity has been observed previously for the maturation-enhancing response in immature aggregate cultures. However, in the present study no correlation was found between the degree of c-fos induction and the extent of the maturation-enhancing stimulation. Immature cultures known to be most sensitive and responsive to these maturation-enhancing agents required relatively high doses of peptide growth factors for the induction of c-fos, and the maximal levels of c-fos mRNA elicited were much lower than those in differentiated cultures which did not show any long-term response to these stimuli.(ABSTRACT TRUNCATED AT 250 WORDS)

Expression of an uncleavable N-terminal RasGAP fragment in insulin-secreting cells increases their resistance toward apoptotic stimuli without affecting their glucose-induced insulin secretion.

Apoptosis of pancreatic beta cells is implicated in the onset of type 1 and type 2 diabetes. Consequently, strategies aimed at increasing the resistance of beta cells toward apoptosis could be beneficial in the treatment of diabetes. RasGAP, a regulator of Ras and Rho GTPases, is an atypical caspase substrate, since it inhibits, rather than favors, apoptosis when it is partially cleaved by caspase-3 at position 455. The antiapoptotic signal generated by the partial processing of RasGAP is mediated by the N-terminal fragment (fragment N) in a Ras-phosphatidylinositol 3-kinase-Akt-dependent, but NF-kappaB-independent, manner. Further cleavage of fragment N at position 157 abrogates its antiapoptotic properties. Here we demonstrate that an uncleavable form of fragment N activates Akt, represses NF-kappaB activity, and protects the conditionally immortalized pancreatic insulinoma betaTC-tet cell line against various insults, including exposure to genotoxins, trophic support withdrawal, and incubation with inflammatory cytokines. Fragment N also induced Akt activity and protection against cytokine-induced apoptosis in primary pancreatic islet cells. Fragment N did not alter insulin cell content and insulin secretion in response to glucose. These data indicate that fragment N protects beta cells without affecting their function. The pathways regulated by fragment N are therefore promising targets for antidiabetogenic therapy.

Examining chemical compound biodegradation at low concentrations through bacterial cell proliferation.

We show proof of principle for assessing compound biodegradation at 1-2 mg C per L by measuring microbial community growth over time with direct cell counting by flow cytometry. The concept is based on the assumption that the microbial community will increase in cell number through incorporation of carbon from the added test compound into new cells in the absence of (as much as possible) other assimilable carbon. We show on pure cultures of the bacterium Pseudomonas azelaica that specific population growth can be measured with as low as 0.1 mg 2-hydroxybiphenyl per L, whereas in mixed community 1 mg 2-hydroxybiphenyl per L still supported growth. Growth was also detected with a set of fragrance compounds dosed at 1-2 mg C per L into diluted activated sludge and freshwater lake communities at starting densities of 10(4) cells per ml. Yield approximations from the observed community growth was to some extent in agreement with standard OECD biodegradation test results for all, except one of the examined compounds.

Contribution à la compréhension du rôle de la mémoire de travail visuospatiale et de ses altérations dans la schizophrénie à l'aide d'un nouveau paradigme qui présente des stimuli dynamiques et statiques

Abstract Working memory has been defined as the ability to maintain and manipulate on-line a limited amount of information. A large number of studies have investigated visuospatial working memory in schizophrenia. However, today, the available data concerning the functioning of these processes in schizophrenia are largely controversial. These inconclusive results are due to incomplete knowledge on processes involved in visuospatial working memory tasks. Recent studies suggest that visuospatial working memory processes may be divided into an active monitoring and a storing components. Furthermore, it has been shown that visuospatial working memory processes are strongly interconnected with early encoding processes (perceptual organization). In our view, the two working memory components, and the relationship that they entertain with early encoding processes, may be investigated using dynamic and static visuospatial stimuli in a working memory paradigm. In this thesis we aim at comparing dynamic and static visuospatial working memory processes in patients with schizophrenia and first-episode of psychosis patients. This analysis may clarify the functioning of visuospatial working memory and the dysfunction of these processes in schizophrenia. Our results are in accord with the hypothesis of two visuospatial working memory subcomponents. Dynamic, rather than static, stimuli are strongly involved in the visuospatial working memory encoding process. Indeed, the results are congruent with the idea that static stimuli may be strongly encoded by parallel perceptual organization processes. Patients with schizophrenia show important deficits in both working memory and perceptual organization encoding processes. In contrast, perceptual organization processes seem spared in firstepisodepsychosis patients. Considering our findings, we propose a model to explain the degradation of visuospatial processes involved in a working memory task during schizophrenia. Résumé: La mémoire de travail est définie comme la capacité à maintenir et manipuler « on-line » un nombre limité d'informations pour une courte période de temps (de l'ordre de quelques dizaines de secondes). Un grand nombre d'études a montré que les processus de mémoire de travail visuo spatiale peuvent être affectés dans la schizophrénie. Malgré cela, les données concernant les déficits de ces processus chez des patients qui souffrent de schizophrénie sont contradictoires. La difficulté de comprendre la nature des déficits de mémoire de travail visuospatiale dans la schizophrénie est en grande partie imputable aux connaissances encore lacunaires sur le fonctionnement de ces processus dans un état non pathologique. Dans cette thèse, on cherche à élucider le fonctionnement des processus de mémoire de travail visuospatiale. Pour cela, on a créé un nouveau paradigme qui sollicite ce type de mémoire. Dans ce paradigme, on présente des stimuli dynamiques et statiques. Après un court délai, le sujet doit reconnaître le stimulus qu'il a visualisé parmi six possibilités. Sur la base de récentes études neurophysiologiques, neuroanatomiques et psychologiques, nous avançons l'hypothèse que l'encodage de stimuli dynamiques et statiques repose sur deux processus distincts de mémoire de travail : un processus d'organisation qui manipule les informations sensorielles et un processus dé stockage qui est responsable du maintien de l'information au cours de la manipulation. Ces processus sont en relation directe avec les processus responsables d'une organisation de l'information à un niveau précoce du traitement visuel. Les études présentées dans cette thèse ont pour but de vérifier la pertinence de la distinction entre les processus de mémoire de travail visuospatiale, selon la modalité «dynamique » ou «statique ». L'investigation des processus dynamiques et statiques de mémoire de travail dans la schizophrénie présente deux avantages principaux. Premièrement, 1a pathologie pourrait permettre de mettre en évidence, par les dysfonctionnements qu'elle présente, la pertinence des hypothèses sur le fonctionnement des processus de mémoire de travail visuospatiale et de leur interaction avec les processus sensoriels. Deuxièmement, ces investigations rendent possible une analyse des dysfonctions des différents processus dans la schizophrénie. Dans cette thèse, on analyse aussi les processus de mémoire de travail «dynamiques » et «statiques » chez des sujets dans une phase initiale de la psychose. Les résultats de cette étude permettent de faire une comparaison avec ceux obtenus avec des patients qui souffrent de schizophrénie. Cette comparaison peut apporter des informations intéressantes sur l'évolution des dysfonctions dans les processus impliqués dans les fonctions de mémoire de travail visuospatiale au cours de la schizophrénie. Les résultats obtenus dans les différentes études sont cohérents avec l'hypothèse d'une implication différente des processus d'organisation de la mémoire de travail sur des stimuli dynamiques et statiques. -Nos résultats montrent que les processus de mémoire de travail responsables de l'organisation (manipulation active) des informations est déficitaire dans la schizophrénie. Ce déficit semble jouer un rôle de premier plan dans la dégradation des processus visuospatiaux au cours de la schizophrénie.

Respiratory responses associated with affective processing of film stimuli

We investigated respiratory responses during film clip viewing and their relation to the affective dimensions of valence and arousal. Seventy-six subjects participated in a study using a between groups design. To begin with, all participants viewed an emotionally neutral film clip. Then, they were presented with one out of four emotional film clips: a positive high-arousal, a positive low-arousal, a negative high-arousal and a negative low-arousal clip. Respiration, skin conductance level, heart rate, corrugator activity and affective judgments were measured. Expiratory time was shorter and inspiratory duty cycle, mean expiratory flow and minute ventilation were larger during the high-arousal clips compared to the low-arousal clips. The pleasantness of the stimuli had no influence on any respiratory measure. These findings confirm the importance of arousal in respiratory responding but also evidence differences in comparison to previous studies using visual and auditory stimuli. [Authors]

Using flow cytometry for in situ monitoring of antimicrobial compound production in the biocontrol bacterium Pseudomonas fluorescens CHA0

Pseudomonas fluorescens strain CHA0 is able to protect plants against a variety of pathogens, notably by producing the two antimicrobial compounds 2,4-diacetylphloroglucinol (DAPG) and pyoluteorin (PLT). The regulation of the expression of these compounds is affected by many biotic factors, such as fungal pathogens, rhizosphere bacteria as well as plant species. Therefore, the influence of some plant phenolic compounds on the expression of DAPG and PLT biosynthetic genes has been tested using GFP-based reporter, monitored by standard fluometry and flow cytometry. In situ experiments were also performed with cucumber plants. We found that several plant metabolites such as IAA and umbelliferone are able to modify significantly the expression of DAPG and PLT. The use of flow cytometry with autofluorescents proteins seems to be a promising method to study rhizobacteria-plant interactions.

A CT-based study investigating the relationship between pedicle screw placement and stimulation threshold of compound muscle action potentials measured by intraoperative neurophysiological monitoring.

PURPOSE: Neurophysiological monitoring aims to improve the safety of pedicle screw placement, but few quantitative studies assess specificity and sensitivity. In this study, screw placement within the pedicle is measured (post-op CT scan, horizontal and vertical distance from the screw edge to the surface of the pedicle) and correlated with intraoperative neurophysiological stimulation thresholds. METHODS: A single surgeon placed 68 thoracic and 136 lumbar screws in 30 consecutive patients during instrumented fusion under EMG control. The female to male ratio was 1.6 and the average age was 61.3 years (SD 17.7). Radiological measurements, blinded to stimulation threshold, were done on reformatted CT reconstructions using OsiriX software. A standard deviation of the screw position of 2.8 mm was determined from pilot measurements, and a 1 mm of screw-pedicle edge distance was considered as a difference of interest (standardised difference of 0.35) leading to a power of the study of 75 % (significance level 0.05). RESULTS: Correct placement and stimulation thresholds above 10 mA were found in 71 % of screws. Twenty-two percent of screws caused cortical breach, 80 % of these had stimulation thresholds above 10 mA (sensitivity 20 %, specificity 90 %). True prediction of correct position of the screw was more frequent for lumbar than for thoracic screws. CONCLUSION: A screw stimulation threshold of >10 mA does not indicate correct pedicle screw placement. A hypothesised gradual decrease of screw stimulation thresholds was not observed as screw placement approaches the nerve root. Aside from a robust threshold of 2 mA indicating direct contact with nervous tissue, a secondary threshold appears to depend on patients' pathology and surgical conditions.

Theoretical approach to the magnetostructural correlations in spin-Peierls compound CuGeO3

A theoretical density-functional study has been carried out to analyze the exchange coupling in the chains of CuGeO3 using discrete models. The results show a good agreement with the experimental exchange coupling constant (J) together with a strong dependence of J with the Cu-O-Cu angle. The calculation of the J values for a distorted model indicates a larger degree of dimerization than those reported previously.

Theoretical approach to the magnetostructural correlations in spin-Peierls compound CuGeO3

A theoretical density-functional study has been carried out to analyze the exchange coupling in the chains of CuGeO3 using discrete models. The results show a good agreement with the experimental exchange coupling constant (J) together with a strong dependence of J with the Cu-O-Cu angle. The calculation of the J values for a distorted model indicates a larger degree of dimerization than those reported previously.

Characterization of PhlG, a hydrolase that specifically degrades the antifungal compound 2,4-diacetylphloroglucinol in the biocontrol agent Pseudomonas fluorescens CHA0.

The potent antimicrobial compound 2,4-diacetylphloroglucinol (DAPG) is a major determinant of biocontrol activity of plant-beneficial Pseudomonas fluorescens CHA0 against root diseases caused by fungal pathogens. The DAPG biosynthetic locus harbors the phlG gene, the function of which has not been elucidated thus far. The phlG gene is located upstream of the phlACBD biosynthetic operon, between the phlF and phlH genes which encode pathway-specific regulators. In this study, we assigned a function to PhlG as a hydrolase specifically degrades DAPG to equimolar amounts of mildly toxic monoacetylphloroglucinol (MAPG) and acetate. DAPG added to cultures of a DAPG-negative DeltaphlA mutant of strain CHA0 was completely degraded, and MAPG was temporarily accumulated. In contrast, DAPG was not degraded in cultures of a DeltaphlA DeltaphlG double mutant. To confirm the enzymatic nature of PhlG in vitro, the protein was histidine tagged, overexpressed in Escherichia coli, and purified by affinity chromatography. Purified PhlG had a molecular mass of about 40 kDa and catalyzed the degradation of DAPG to MAPG. The enzyme had a kcat of 33 s(-1) and a Km of 140 microM at 30 degrees C and pH 7. The PhlG enzyme did not degrade other compounds with structures similar to DAPG, such as MAPG and triacetylphloroglucinol, suggesting strict substrate specificity. Interestingly, PhlG activity was strongly reduced by pyoluteorin, a further antifungal compound produced by the bacterium. Expression of phlG was not influenced by the substrate DAPG or the degradation product MAPG but was subject to positive control by the GacS/GacA two-component system and to negative control by the pathway-specific regulators PhlF and PhlH.

Optimal dividend strategies for a compound Poisson risk process under transaction costs and power utility

We characterize the value function of maximizing the total discounted utility of dividend payments for a compound Poisson insurance risk model when strictly positive transaction costs are included, leading to an impulse control problem. We illustrate that well known simple strategies can be optimal in the case of exponential claim amounts. Finally we develop a numerical procedure to deal with general claim amount distributions.