Glutathione deficit in schizophrenia: strategies to increase glutathione levels in " in vitro " and clinical studies

Summary: Decrease in glutathione (GSH) levels was observed in cerebrospinal fluid, prefrontal cortex and post-mortem striatum of schizophrenia patients. Evidences suggest a defect in GSH synthesis at the levels of the rate-limiting synthesizing enzyme, glutamate cysteine ligase (GCL). Indeed, polymorphisms in the gene of the modifier subunit of GCL (GCLM) was shown to be associated with the disease in three different populations, GCLM gene expression is decreaséd in fibroblasts from patients and the increase in GCL activity induced by an oxidative stress is lower in patients' fibroblasts compared to controls. GSH being a major antioxydant and redox regulator, its presence is of high importance for protecting cells against oxidative stress. The aim of the present work was to use various substances to increase GSH levels by diverse strategies. Since the synthesizing enzyme GCL is defective, bypassing this enzyme was the first strategy we used. GSH ethyl ester (GSHEE), a membrane permeable analog of GSH, succeeded in replenishing GSH levels in cultured neurons and astrocytes previously depleted in GSH by L-buthionine-(S,R)-sulfoximine (BSO), an inhibitor of GCL. GSHEE also abolished dopamine-induced decrease of NMDA-mediated calcium response observed in BSO-treated neurons. y-Glutamylcysteine ethyl ester (GCSE), a membrane permeable analog of the product of GCL, increased GSH levels only in astrocytes. The second strategy was to boost the defective enzyme GCL. While quercetin (flavonoid) could increase GSH levels only in astrocytes, curcumin (polyphenol) and tertbutylhydroquinone (quinone) were successful in both neurons and astrocytes, via an increase in the gene expression of the two subunits of GCL and, consequently, an increase in the activity of the enzyme. However, FK506, an immunosupressant, was unefficient. Treating astrocytes from GCLM KO mice showed that the modulatory subunit is necessary for the action of the substances. Finally, since cysteine is the limiting precursor in the synthesis of GSH, we hypothesized that we could increase GSH levels by providing more of this precursor. N-acetyl-cysteine (NAC), a cysteine donor, was administered to schizophrenia patients, using adouble-blind and cross-over protocol. NAC significantly improved the mismatch negativity (MMN), a component of the auditory evoked potentials, thought to reflect selective current flowing through open, unblocked NMDA channels. Considering that NMDA function is reduced when GSH levels are low, increasing these levels with NAC could improve NMDA function as reflected by the improvement in the generation of the MMN. Résumé: Les taux de glutathion (GSH) dans le liquide céphalo-rachidien, le cortex préfrontal ainsi que le striatum post-mortem de patients schizophrènes, sont diminués. L'enzyme limitante dans la synthèse du GSH, la glutamyl-cysteine ligase (GCL), est défectueuse. En effet, des polymorphismes dans le gène de la sous-unité modulatrice de GCL (GCLM) sont associés à la maladie, l'expression du gène GCLM est diminuée dans les fibroblastes de patients et, lors d'un stress oxidative, l'augmentation de l'activité de GCL est plus faible chez les patients que chez les contrôles. Le GSH étant un important antioxydant et régulateur du status redox, sa présence est primordiale afin de protéger les cellules contre les stress oxydatifs. Au cours du présent travail, une variété de substances ont été utilisées dans le but d'augmenter les taux de GSH. Passer outre l'enzyme de synthèse GCL qui est défectueuse fut la première stratégie utilisée. L'éthylester de GSH (GSHEE), un analogue du GSH qui pénètre la membrane cellulaire, a augmenté les taux de GSH dans des neurones et des astrocytes déficitaires en GSH dû au L-buthionine-(S,R)-sulfoximine (BSO), un inhibiteur du GCL. Dans ces neurones, le GSHEE a aussi aboli la diminution de la réponse NMDA, induite parla dopamine. L'éthyl-ester de y-glutamylcysteine (GCEE), un analogue du produit de la GCL qui pénètre la membrane cellulaire, a augmenté les taux de GSH seulement dans les astrocytes. La seconde stratégie était d'augmenter l'activité de l'enzyme GCL. Tandis que la quercétine (flavonoïde) n'a pu augmenter les taux de GSH que dans les astrocytes, la curcumin (polyphénol) et le tert-butylhydroquinone (quinone) furent efficaces dans les deux types de cellules, via une augmentation de l'expression des gènes des deux sous-unités de GCL et de l'activité de l'enzyme. Le FK506 (immunosupresseur) n' a démontré aucune efficacité. Traiter des astrocytes provenant de souris GCLM KO a permis d'observer que la sous-unité modulatoire est nécessaire à l'action des substances. Enfin, puisque la cysteine est le substrat limitant dans la synthèse du GSH, fournir plus de ce présurseur pourrait augmenter les taux de GSH. Nacétyl-cystéine (NAC), un donneur de cystéine, a été administrée à des schizophrènes, lors d'une étude en double-aveugle et cross-over. NAC a amélioré le mismatch negativity (MMN), un composant des potentials évoqués auditifs, qui reflète le courant circulant via les canaux NMDA. Puisque la fonctionnalité des R-NMDA est diminuée lorsque les taux de GSH sont bas, augmenter ces taux avec NAC pourrait améliorer la fonction des R-NMDA, réflété par une augmentation de l'amplitude du MMN.

Design of axial-flux permanent-magnet low-speed machines and performance comparison between radial-flux and axial-flux machines

This thesis presents an alternative approach to the analytical design of surface-mounted axialflux permanent-magnet machines. Emphasis has been placed on the design of axial-flux machines with a one-rotor-two-stators configuration. The design model developed in this study incorporates facilities to include both the electromagnetic design and thermal design of the machine as well as to take into consideration the complexity of the permanent-magnet shapes, which is a typical requirement for the design of high-performance permanent-magnet motors. A prototype machine with rated 5 kW output power at 300 min-1 rotation speed has been designed and constructed for the purposesof ascertaining the results obtained from the analytical design model. A comparative study of low-speed axial-flux and low-speed radial-flux permanent-magnet machines is presented. The comparative study concentrates on 55 kW machines with rotation speeds 150 min-1, 300 min-1 and 600 min-1 and is based on calculated designs. A novel comparison method is introduced. The method takes into account the mechanical constraints of the machine and enables comparison of the designed machines, with respect to the volume, efficiency and cost aspects of each machine. It is shown that an axial-flux permanent-magnet machine with one-rotor-two-stators configuration has generally a weaker efficiency than a radial-flux permanent-magnet machine if for all designs the same electric loading, air-gap flux density and current density have been applied. On the other hand, axial-flux machines are usually smaller in volume, especially when compared to radial-flux machines for which the length ratio (axial length of stator stack vs. air-gap diameter)is below 0.5. The comparison results show also that radial-flux machines with alow number of pole pairs, p < 4, outperform the corresponding axial-flux machines.

REGSTATTOOLS: freeware statistical tools for the analysis of disease population databases used in health and social studies

Background: The repertoire of statistical methods dealing with the descriptive analysis of the burden of a disease has been expanded and implemented in statistical software packages during the last years. The purpose of this paper is to present a web-based tool, REGSTATTOOLS http://regstattools.net intended to provide analysis for the burden of cancer, or other group of disease registry data. Three software applications are included in REGSTATTOOLS: SART (analysis of disease"s rates and its time trends), RiskDiff (analysis of percent changes in the rates due to demographic factors and risk of developing or dying from a disease) and WAERS (relative survival analysis). Results: We show a real-data application through the assessment of the burden of tobacco-related cancer incidence in two Spanish regions in the period 1995-2004. Making use of SART we show that lung cancer is the most common cancer among those cancers, with rising trends in incidence among women. We compared 2000-2004 data with that of 1995-1999 to assess percent changes in the number of cases as well as relative survival using RiskDiff and WAERS, respectively. We show that the net change increase in lung cancer cases among women was mainly attributable to an increased risk of developing lung cancer, whereas in men it is attributable to the increase in population size. Among men, lung cancer relative survival was higher in 2000-2004 than in 1995-1999, whereas it was similar among women when these time periods were compared. Conclusions: Unlike other similar applications, REGSTATTOOLS does not require local software installation and it is simple to use, fast and easy to interpret. It is a set of web-based statistical tools intended for automated calculation of population indicators that any professional in health or social sciences may require.

Making Space for Festival 1400-1700. Interactions of Architecture and Performance in Late Medieval and Early Modern Festivals

Las relaciones entre actos festivos y espacios arquitectónicos, y las modalidades con que las ceremonias cortesanas llegaban a modificar los centros urbanos y las residen- cias palaciegas, han sido los temas principales del congreso internacional Making Space for Festival, 14001700. Interactions of Architecture and Performance in Late Medieval and Early Modern Festivals, que tuvo lugar del 21 al 24 de marzo de 2013 en el Palacio Pesaro-Papafava de Venecia, residencia de la Universidad de Warwick en la Laguna. La Society for European Festivals Research (SEFR), entidad vinculada a esta universidad, ha promovido el encuentro junto con la Red PALATIUM financia- da por el Research Networking Programme de la European Science Fundation (ESF) (2010-2015), dedicada al estudio de las residencias palaciegas de la Edad Moderna como espacios de intercambios políticos y culturales. El tema de las jornadas nacía de la unión de los intereses científicos de los grupos organizadores, dejando lugar a temáticas diferentes como: la articulación de un mensaje festivo a lo largo del recorri- do de procesiones y entradas públicas, la financiación y los aspectos más prácticos de la puesta en escena de un evento festivo, la tipificación de ceremonias y decoraciones en determinados contextos, el intercambio cultural a través de la fiesta, o la recepción de las modificaciones de los espacios por parte de los espectadores

Load and Performance Testing of Web Applications on Service Based Environments

Suorituskyky- ja kuormitustestien tekeminen sovelluksille on erittäin tärkeä osa tuotantoprosessia nykypäivänä. Myös Web-sovelluksia testataan yhä enemmän. Tarve suorituskyky- ja kuormitustestien tekemiselle on selvä. Testattavan ympäristön tämänhetkinen, mutta myös tulevaisuuden toimivuus taataan oikein tehdyillä testeillä ja niitä seuraavilla korjaustoimenpiteillä. Suurten käyttäjämäärien testaaminen manuaalisesti on kuitenkin hyvin vaikeaa. Sirpaleisen ympäristön, kuten palveluihin perustuvien Web-sovellusympäristöjen testaaminen on haaste. Tämän työn aiheena on arvioida työkaluja ja menetelmiä, joilla raskaita teollisia Web-sovelluksia voidaan testata. Tavoitteena on löytää testausmenetelmiä, joilla voidaan luotettavasti simuloida suuria käyttäjämääriä. Tavoitteena on myös arvioida erilaisten yhteyksien ja protokollien vaikutusta Web-sovelluksen suorituskykyyn.

Évaluation d'une structure gériatrique entre l'ambulatoire et l'hospitalier [Geriatric institutions, between ambulatory and hospital care: patients' description and performance assessment].

Introduction : Décrire les patients d'une structure gériatrique offrant des hospitalisations de courte durée, dans un contexte ambulatoire, pour des situations gériatriques courantes dans le canton de Genève (Suisse). Mesurer les performances de cette structure en termes de qualité des soins et de coûts. Méthodes : Des données relatives au profil des 100 premiers patients ont été collectées (huit mois), ainsi qu'aux prestations, aux ressources et aux effets (réadmissions, décès, satisfaction, complications) de manière à mesurer différents indicateurs de qualité et de coûts. Les valeurs observées ont été systématiquement comparées aux valeurs attendues, calculées à partir du profil des patients. Résultats : Des critères d'admission ont été fixés pour exclure les situations dans lesquelles d'autres structures offrent des soins mieux adaptés. La spécificité de cette structure intermédiaire a été d'assurer une continuité des soins et d'organiser d'emblée le retour à domicile par des prestations de liaison ambulatoire. La faible occurrence des réadmissions potentiellement évitables, une bonne satisfaction des patients, l'absence de décès prématurés et le faible nombre de complications suggèrent que les soins médicaux et infirmiers ont été délivrés avec une bonne qualité. Le coût s'est révélé nettement plus économique que des séjours hospitaliers après ajustement pour la lourdeur des cas. Conclusion : L'expérience-pilote a démontré la faisabilité et l'utilité d'une unité d'hébergement et d'hospitalisation de court séjour en toute sécurité. Le suivi du patient par le médecin traitant assure une continuité des soins et évite la perte d'information lors des transitions ainsi que les examens non pertinents. INTRODUCTION: To describe patients admitted to a geriatric institution, providing short-term hospitalizations in the context of ambulatory care in the canton of Geneva. To measure the performances of this structure in terms of quality ofcare and costs. METHOD: Data related to the clinical,functioning and participation profiles of the first 100 patients were collected. Data related to effects (readmission, deaths, satisfaction, complications), services and resources were also documented over an 8-month period to measure various quality and costindicators. Observed values were systematically compared to expected values, adjusted for case mix. RESULTS: Explicit criteria were proposed to focus on the suitable patients, excluding situations in which other structures were considered to be more appropriate. The specificity of this intermediate structure was to immediately organize, upon discharge, outpatient services at home. The low rate of potentially avoidable readmissions, the high patient satisfaction scores, the absence of premature death and the low number of iatrogenic complications suggest that medical and nursing care delivered reflect a good quality of services. The cost was significantly lower than expected, after adjusting for case mix. CONCLUSION: The pilot experience showed that a short-stay hospitalization unit was feasible with acceptable security conditions. The attending physician's knowledge of the patients allowed this system tofocus on essential issues without proposing inappropriate services.

Influence of Equipment Operation and Performance Deterioration on Power Plant Economy.

Pysyäkseen kilpailukykyisenä vapautuneilla sähkömarkkinoilla on voimalaitoksen energiantuotantokustannusten oltava mahdollisimman matalia, tinkimättä kuitenkaan korkeasta käytettävyydestä. Polttoaineen energiasisällön mahdollisimman hyvä hyödyntäminen on ratkaisevan tärkeää voimalaitoksen kannattavuudelle. Polttoainekustannusten osuus on konvektiivisilla laitoksilla yleensä yli puolet koko elinjakson kustannuksista. Kun vielä päästörajat tiukkenevat koko ajan, korostuu polttoaineen korkea hyötykäyttö entisestään. Korkea energiantuotannon luotettavuus ja käytettävyys ovat myös elintärkeitä pyrittäessä kustannusten minimointiin. Tässä työssä on käyty läpi voimalaitoksen kustannuksiin vaikuttavia käsitteitä, kuten hyötysuhdetta, käytettävyyttä, polttoaineen hintoja, ylös- ja alasajoja ja tärkeimpiä häviöitä. Ajostrategiassa ja poikkeamien hallinnassa pyritään hyvään hyötysuhteeseen ja alhaisiin päästöihin joka käyttötilanteessa. Lisäksi on tarkasteltu tiettyjen suureiden, eli höyryn lämpötilan ja paineen, savukaasun hapen pitoisuuden, savukaasun loppulämpötilan, sekä lauhduttimen paineen poikkeamien vaikutusta ohjearvostaan energiantuotantokustannuksiin. Happi / hiilimonoksidi optimoinnissa on otettu huomioon myös pohjatuhkan palamattomat.

Development, physical-chemical stability and release studies of four alcohol-free spironolactone suspensions for use in pediatrics

Dissolution studies have become of great significance because, in most cases, drug dissolution is the rate-limiting step in the absorption process. As occurs with solid oral dosage forms, heterogeneous disperse systems (suspensions) could also have some problems with their in vitro dissolution. The objective of this study was to evaluate influence of the excipients on the release of spironolactone from four alcohol free suspensions (pharmaceutical compounding) of spironolactone 5 mg/mL suitable for pediatric use. Also the comparison of the physical and chemical stability of the suspensions stored at 4, 25 and 40 ºC over a 60- day period has been studied. Rheological behavior, particle size, a prediction of long-term physical stability, pH and assay of spironolactone by HPLC were assessed at prefixed times. The dissolution profile of each suspension was determined and compared with that of the commercial tablets. A microbiological study of the best formula was also performed. Chemically, the four spironolactone suspensions were stable for 60 days stored at three temperatures; Suspension IV had optimum pH values and the highest recovery percentage. In terms of physical stability, sedimentation occurred in Suspension IV and flotation of spironolactone in Suspensions I, II and III. Suspension III had the highest viscosity and the slowest drug release. Suspension IV was also microbiologically stable for 60 days. In conclusion, Suspension IV had the best properties and the least suitable form was Suspension III, as its high viscosity made it difficult to achieve homogeneous redispersion, and it had the slowest dissolution profile.

A chromosomal insertion toolbox for promoter probing, mutant complementation and pathogenicity studies in Ralstonia solanacearum

We describe here the construction of a delivery system for stable and directed insertion of gene constructs in a permissive chromosomal site of the bacterial wilt pathogen Ralstonia solanacearum. The system consists of a collection of suicide vectors the Ralstonia chromosome (pRC) series that carry an integration element flanked by transcription terminators and two sequences of homology to the chromosome of strain GMI1000, where the integration element is inserted through a double recombination event. Unique restriction enzyme sites and a GATEWAY cassette enable cloning of any promoter::gene combination in the integration element. Variants endowed with different selectable antibiotic resistance genes and promoter::gene combinations are described. We show that the system can be readily used in GMI1000 and adapted to other R. solanacearum strains using an accessory plasmid. We prove that the pRC system can be employed to complement a deletion mutation with a single copy of the native gene, and to measure transcription of selected promoters in monocopy both in vitro and in planta. Finally, the system has been used to purify and study secretion type III effectors. These novel genetic tools will be particularly useful for the construction of recombinant bacteria that maintain inserted genes or reporter fusions in competitive situations (i.e., during plant infection).

Market orientation and performance in entrepreneurial SMEs

Market orientation is the organizational culture that creates the necessary behaviors for continuous additional value for customers and thus continuous superior performance for the business. The field of market orientation has been studied repeatedly during the past two decades. Yet research has concentrated on large firms in large domestic markets creating a need for diversifying research. The master’s thesis at hand examined the general incidence of market orientation among SMEs from five different industries as well as its consequences on SME performance. The empirical part of the thesis was conducted with a web-based survey that resulted in 255 responses. The data of the survey was analyzed by statistical analysis. The incidence of market orientation varied among dimensions and market orientation did not show any direct effect on firm performance. Customer orientation was the only dimension that showed a direct (positive) effect. On the contrary, moderating effects were found which indicate that the effect of market orientation in SMEs is influenced by other factors that should receive further attention. Also industry specific differences were discovered and should be further examined.

Strategic groups and performance: an empirical analysis of strategic groups in the wind energy industry in Europe

The strategic group theory provides an intermediate level of analysis between a single company and the whole industry for identifying issues about the company's competitive position and strategic choices. Strategic groups are companies within an industry with similar strategic characteristics or competing on similar bases. Strategic choices are aligned with the firms’ resources. The purpose of this study was to identify the strategic groups in the wind energy industry in Europe, and study, whether a certain group membership results in financial performance differences. Altogether 80 European wind energy companies were included in the study, which were clustered into four strategic groups according to their age and growth rate. Each group corresponds to a different strategy. The results show that the wind energy companies can be clustered according to the chosen strategic characteristics. Strategic decisions were investigated with characteristic variables. Performance variables were used in the analysis measuring profitability, liquidity and solvency of the groups. These strategic choices of the companies did not have a significant influence on the firms’ performance. The more mature and slower growing group proved to be the most successful. However, the differences between groups were generally not statistically significant. The only statistically significant difference found was in the solvency ratio between Mature Slow and Young Rapid groups. Measured with these variables, more mature and slower growing companies performed better. Therefore, a certain strategic group membership results in performance differences.