An Atypical Mitochondrial Carrier That Mediates Drug Action in Trypanosoma brucei

Elucidating the mechanism of action of trypanocidal compounds is an important step in the development of more efficient drugs against Trypanosoma brucei. In a screening approach using an RNAi library in T. brucei bloodstream forms, we identified a member of the mitochondrial carrier family, TbMCP14, as a prime candidate mediating the action of a group of anti-parasitic choline analogs. Depletion of TbMCP14 by inducible RNAi in both bloodstream and procyclic forms increased resistance of parasites towards the compounds by 7-fold and 3-fold, respectively, compared to uninduced cells. In addition, down-regulation of TbMCP14 protected bloodstream form mitochondria from a drug-induced decrease in mitochondrial membrane potential. Conversely, over-expression of the carrier in procyclic forms increased parasite susceptibility more than 13-fold. Metabolomic analyses of parasites over-expressing TbMCP14 showed increased levels of the proline metabolite, pyrroline-5-carboxylate, suggesting a possible involvement of TbMCP14 in energy production. The generation of TbMCP14 knock-out parasites showed that the carrier is not essential for survival of T. brucei bloodstream forms, but reduced parasite proliferation under standard culture conditions. In contrast, depletion of TbMCP14 in procyclic forms resulted in growth arrest, followed by parasite death. The time point at which parasite proliferation stopped was dependent on the major energy source, i.e. glucose versus proline, in the culture medium. Together with our findings that proline-dependent ATP production in crude mitochondria from TbMCP14-depleted trypanosomes was reduced compared to control mitochondria, the study demonstrates that TbMCP14 is involved in energy production in T. brucei. Since TbMCP14 belongs to a trypanosomatid-specific clade of mitochondrial carrier family proteins showing very poor similarity to mitochondrial carriers of mammals, it may represent an interesting target for drug action or targeting.

Does the use of mobile devices (tablets and smartphones) affect survey quality and choice behaviour in web surveys?

Web surveys are becoming increasingly popular in survey research including stated preference surveys. Compared with face-to-face, telephone and mail surveys, web surveys may contain a different and new source of measurement error and bias: the type of device that respondents use to answer the survey questions. This is the first study that tests whether the use of mobile devices, tablets or smartphones, affects survey characteristics and stated preferences in a web-based choice experiment. The web survey on expanding renewable energy production in Germany was carried out with 3182 respondents, of which 12% used a mobile device. Propensity score matching is used to account for selection bias in the use of mobile devices for survey completion. We find that mobile device users spent more time than desktop/laptop users to answer the survey. Yet, desktop/laptop users and mobile device users do not differ in acquiescence tendency as an indicator of extreme response patterns. For mobile device users only, we find a negative correlation between screen size and interview length and a positive correlation between screen size and acquiescence tendency. In the choice experiment data, we do not find significant differences in the tendency to choose the status quo option and scale between both subsamples. However, some of the estimates of implicit prices differ, albeit not in a unidirectional fashion. Model results for mobile device users indicate a U-shaped relationship between error variance and screen size. Together, the results suggest that using mobile devices is not detrimental to survey quality.

Anoxia tolerance in tobacco roots: effect of overexpression of pyruvate decarboxylase

Plant survival during flooding relies on ethanolic fermentation for energy production. The available literature indicates that the first enzyme of the ethanolic fermentation pathway, pyruvate decarboxylase (PDC), is expressed at very low levels and is likely to be rate-limiting during oxygen deprivation. The authors expressed high levels of bacterial PDC in tobacco to study the modulation of PDC activity in vivo, and assess its impact on the physiology of ethanolic fermentation and survival under oxygen stress. In contrast to leaves, wild-type normoxic roots contained considerable PDC activity, and overexpression of the bacterial PDC caused only a moderate increase in acetaldehyde and ethanol production under anoxia compared to wild-type roots. No significant lactate production could be measured at any time, making it unlikely that lactate-induced acidification (LDH/PDC pH-stat) triggers the onset of ethanol synthesis. Instead, the authors favour a model in which the flux through the pathway is regulated by substrate availability. The increased ethanolic flux in the transgenics compared to the wild-type did not enhance anoxia tolerance. On the contrary, rapid utilisation of carbohydrate reserves enhanced premature cell death in the transgenics while replenishment of carbohydrates improved survival under anoxia.

LOW REACTIVE OXYGEN SPECIES AND HIGH GLYCOLYSIS IN GLIOBLASTOMA STEM CELLS:MECHANISMS AND THERAPEUTIC IMPLICATIONS

Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumor with poor prognosis due in part to drug resistance and high incidence of tumor recurrence. The drug resistant and cancer recurrence phenotype may be ascribed to the presence of glioblastoma stem cells (GSCs), which seem to reside in special stem-cell niches in vivo and require special culture conditions including certain growth factors and serum-free medium to maintain their stemness in vitro. Exposure of GSCs to fetal bovine serum (FBS) can cause their differentiation, the underlying mechanism of which remains unknown. Reactive oxygen species (ROS) play an important role in normal stem cell differentiation, but their role in affecting cancer stem cell fate remains unclear. Whether the metabolic characteristics of GSCs are different from other glioblastoma cells and can be targeted are also unknown. In this study, we used several stem-like glioblastoma cell lines derived from clinical tissues by typical neurosphere culture system or orthotopic xenografts, and showed that addition of fetal bovine serum to the medium induced an increase of ROS, leading to aberrant differentiation and decreases of stem cell markers such as CD133. We found that exposure of GSCs to serum induced their differentiation through activation of mitochondrial respiration, leading to an increase in superoxide (O2-) generation and a profound ROS stress response manifested by upregulation of oxidative stress response pathway. This increase in mitochondrial ROS led to a down-regulation of molecules including SOX2, and Olig2, and Notch1 that are important for stem cell function and an upregulation of mitochondrial superoxide dismutase SOD2 that converts O2- to H2O2. Neutralization of ROS by antioxidant N-acetyl-cysteine in the serum-treated GSCs suppressed the increase of superoxide and partially rescued the expression of SOX2, Olig2, and Notch1, and prevented the serum-induced differentiation phenotype. Additionally, GSCs showed high dependence on glycolysis for energy production. The combination of a glycolytic inhibitor 3-BrOP and a chemotherapeutic agent BCNU depleted cellular ATP and inhibited the repair of BCNU-induced DNA damage, achieving strikingly synergistic killing effects in drug resistant GSCs. This study uncovers the metabolic properties of glioblastoma stem cells and suggests that mitochondrial function and cellular redox status may profoundly affect the fates of glioblastoma stem cells via a ROS-mediated mechanism, and that the active glycolytic metabolism in cancer stem cells may provide a biochemical basis for developing novel therapeutic strategies to effectively eliminate GSCs.

Exploring the Impact of Decreased AMPK Pathway Activity on Brain Injury Outcome

Each year, 150 million people sustain a Traumatic Brain Injury (TBI). TBI results in life-long cognitive impairments for many survivors. One observed pathological alteration following TBI are changes in glucose metabolism. Altered glucose uptake occurs in the periphery as well as in the nervous system, with an acute increase in glucose uptake, followed by a prolonged metabolic suppression. Chronic, persistent suppression of brain glucose uptake occurs in TBI patients experiencing memory loss. Abberant post-injury activation of energy-sensing signaling cascades could result in perturbed cellular metabolism. AMP-activated kinase (AMPK) is a kinase that senses low ATP levels, and promotes efficient cell energy usage. AMPK promotes energy production through increasing glucose uptake via glucose transporter 4 (GLUT4). When AMPK is activated, it phosphorylates Akt Substrate of 160 kDa (AS160), a Rab GTPase activating protein that controls Glut4 translocation. Additionally, AMPK negatively regulates energy-consumption by inhibiting protein synthesis via the mechanistic Target of Rapamycin (mTOR) pathway. Given that metabolic suppression has been observed post-injury, we hypothesized that activity of the AMPK pathway is transiently decreased. As AMPK activation increases energy efficiency of the cell, we proposed that increasing AMPK activity to combat the post-injury energy crisis would improve cognitive outcome. Additionally, we expected that inhibiting AMPK targets would be detrimental. We first investigated the role of an existing state of hyperglycemia on TBI outcome, as hyperglycemia correlates with increased mortality and decreased cognitive outcome in clinical studies. Inducing hyperglycemia had no effect on outcome; however, we discovered that AMPK and AS160 phosphorylation were altered post-injury. We conducted vii work to characterize this period of AMPK suppression and found that AMPK phosphorylation was significantly decreased in the hippocampus and cortex between 24 hours and 3 days post-injury, and phosphorylation of its downstream targets was consistently altered. Based on this period of observed decreased AMPK activity, we administered an AMPK activator post-injury, and this improved cognitive outcome. Finally, to examine whether AMPK-regulated target Glut4 is involved in post-injury glucose metabolism, we applied an inhibitor and found this treatment impaired post-injury cognitive function. This work is significant, as AMPK activation may represent a new TBI therapeutic target.

Characterization of the NOP -1 opsin photoreceptor in the filamentous fungus Neurospora crassa

Light absorption is an important process for energy production and sensory perception in many organisms. In the filamentous fungus, Neurospora crassa, blue-light is an important regulator of both asexual and sexual development, but the identity of the blue-light receptor is unknown. The work presented in this dissertation initiated the characterization of the putative N. crassa opsin photoreceptor, NOP-1. Opsins were thought to exist only in the archaea and mammals until the discovery of nop-1. All opsins have the same conserved structure of seven transmembrane helical domains with a lysine residue in the seventh helix specific for forming a Schiff-base linkage with retinal. The predicted NOP-1 protein sequence is equally similar to archaeal rhodopsins and a newly identified fungal opsin-related protein group (ORPs). ORPs maintain the seven transmembrane helical structure of opsins, but lack the conserved lysine residue for binding retinal. An ORP gene, orp-1 was identified in N. crassa and this work includes the cloning and sequence analysis of this gene. Characterization of NOP-1 function in N. crassa development began with the construction of a Δnop-1 deletion mutant. Extensive phenotypic analysis of Δnop-1 mutants revealed only subtle defects during development primarily under environmental conditions that induce a stress response. NOP-1 was overexpressed in the heterologous system Pichia pastoris, and it was demonstrated that NOP-1 protein bound all-trans retinal to form a green-light absorbing pigment (λmax = 534 nm) with a photochemical reaction cycle similar to archaeal sensory rhodopsins. nop-1 gene expression was monitored during N. crassa development. nop-1 transcript is highly expressed during asexual sporulation (conidiation) and transcript levels are abundant in the later stages of conidial development. nop-1 expression is not regulated by blue-light or elevated temperatures. Potential functions for NOP-1 were discovered through the transcriptional analysis of conidiation-associated genes in Δnop-1 mutants. NOP-1 exhibits antagonistic transcriptional regulation of conidiation-associated genes late in conidial development, by enhancing the carotenogenic gene, al-2 and repressing the conidiation-specific genes, con-10 and con-13. ^

Euphausiid respiration model revamped, link to model results

Euphausiids constitute major biomass component in shelf ecosystems and play a fundamental role in the rapid vertical transport of carbon from the ocean surface to the deeper layers during their daily vertical migration (DVM). DVM depth and migration patterns depend on oceanographic conditions with respect to temperature, light and oxygen availability at depth, factors that are highly dependent on season in most marine regions. Changes in the abiotic conditions also shape Euphausiid metabolism including aerobic and anaerobic energy production. Here we introduce a global krill respiration model which includes the effect of latitude (LAT), the day of the year of interest (DoY), and the number of daylight hours on the day of interest (DLh), in addition to the basal variables that determine ectothermal oxygen consumption (temperature, body mass and depth) in the ANN model (Artificial Neural Networks). The newly implemented parameters link space and time in terms of season and photoperiod to krill respiration. The ANN model showed a better fit (r**2=0.780) when DLh and LAT were included, indicating a decrease in respiration with increasing LAT and decreasing DLh. We therefore propose DLh as a potential variable to consider when building physiological models for both hemispheres. We also tested for seasonality the standard respiration rate of the most common species that were investigated until now in a large range of DLh and DoY with Multiple Linear Regression (MLR) or General Additive model (GAM). GAM successfully integrated DLh (r**2= 0.563) and DoY (r**2= 0.572) effects on respiration rates of the Antarctic krill, Euphausia superba, yielding the minimum metabolic activity in mid-June and the maximum at the end of December. Neither the MLR nor the GAM approach worked for the North Pacific krill Euphausia pacifica, and MLR for the North Atlantic krill Meganyctiphanes norvegica remained inconclusive because of insufficient seasonal data coverage. We strongly encourage comparative respiration measurements of worldwide Euphausiid key species at different seasons to improve accuracy in ecosystem modelling.

Case studies of the effects of a total solar eclipse on weather with varying degrees of cloud cover using a local numerical weather prediction model

The importance of renewable energies for the European electricity market is growing rapidly. This presents transmission grids and the power market in general with new challenges which stem from the higher spatiotemporal variability of power generation. This uncertainty is due to the fact that renewable power production results from weather phenomena, thus making it difficult to plan and control. We present a sensitivity study of a total solar eclipse in central Europe in March. The weather in Germany and Europe was modeled using the German Weather Service's local area models COSMO-DE and COSMO-EU, respectively (http://www.cosmo-model.org/). The simulations were performed with and without considering a solar eclipse for the following 3 situations: 1. An idealized, clear-sky situation for the entire model area (Europe, COSMO-EU) 2. A real weather situation with mostly cloudy skies (Germany, COSMO-DE) 3. A real weather situation with mostly clear skies (Germany, COSMO-DE) The data should help to evaluate the effects of a total solar eclipse on the weather in the planetary boundary layer. The results show that a total solar eclipse has significant effects particularly on the main variables for renewable energy production, such as solar irradiation and temperature near the ground.

Table 6 Generalised properties of Utsira caprocks, based on analysis of cuttings (page 79)

Executive Summary: Carbon dioxide capture and storage (CCS) is one option for mitigating atmospheric emissions of carbon dioxide and thereby contributes in actions for stabilization of atmospheric greenhouse gas concentrations. The Bellona Foundation is striving to achieve wide implementation of carbon dioxide (CO2) capture and storage both in Norway and internationally. Bellona considers CCS as the only viable large scale option to close the gap between energy production and demand in an environmentally sound way, thereby ensuring that climate changes and acidification of the oceans due to increased CO2 concentrations in the atmosphere will be stabilised. ff

Infrastructure Development of Railway in Cambodia: A Long Term Strategy

Infrastructure development means for the making of living environment, transport and communications, disaster prevention and national land conservation, agriculture, forestry and fisheries, and energy production and supply. Transport infrastructure development in Cambodia involved with (1) road, (2) railway, (3) port, inland-water way and (4) aviation. All model of transport infrastructure have special different kinds of importance. Railway is different from other base important of railways are transport passengers and traffic freight especially transport for heavy goods in huge capacity and in long distance by safer and faster. Transport in Cambodia for traffic freight export import base from Thailand and other via Sisophon and Shihanoukvill port. Traffic is increasing rapidly during nowadays railway condition in adequate of demand required. This is why Railway is selected as the topic of this paper to prevent monopoly of road transport. This paper, does review about infrastructure development plan for Railway in Cambodia as a long term strategy by review and analysis forecast on the previous performance of Royal Railways of Cambodia (RRC) transport traffic involved with condition of infrastructure development of railway in Cambodia. And also review the plan of development RRC but just only detail a plan of rehabilitation that is immediately needed. Suggest some recommendation at the last part. As Cambodia is a member country of ASEAN and also Mekong sub-region. For make sure that transport networks work effectively with a progress of economic integration, we make clear what is important for infrastructure development of railway in Cambodia from the standpoint of the development plan of Mekong sub-region. This paper is organized by 4 sections. Section 1 review about Infrastructure Development of Railway in Cambodia (IDRC) Historical Background, Follow by Section 2 will review the Current Situation of IDRC and some analysis of transport performance from previous years, Then Section 3 review of the focusing on traffic transport of RRC in the future, Section 4 review Infrastructure Development of Railway in Cambodia Future plans in long term; at last conclusion and recommendation. In section 1 does review history background of RRC from the rail first begun. But why is needed to review? Because of history background is involved infrastructure development of RRC in present time. History background made big gaps constraint and obstacle for socioeconomic development and poverty reduction, also left Cambodia with tragedy and left developed behind. After that remain infrastructure development needs huge fund and long time for restoration, reconstruction, rehabilitation and development into new technology as most of world practice.

High-efficiency Köhler photovoltaic concentrators with external light confinement

Metal grid lines are a vital element in multijunction solar cells in order to take out from the cell the generated photocurrent. Nevertheless all this implies certain shadowing factor and thus certain reflectivity on cells surface that lowers its light absorption. This reflectivity produces a loss in electrical efficiency and thus a loss in global energy production for CPV systems. We present here an optical design for recovering this portion of reflected light, and thus leading to a system efficiency increase. This new design is based on an external confinement cavity, an optical element able to redirect the light reflected by the cell towards its surface again. It has been possible thanks to the recent invention of the advanced Köhler concentrators by LPI, likely to integrate one of these cavities easily. We have proven the excellent performance of these cavities integrated in this kind of CPV modules offering outstanding results: 33.2% module electrical efficiency @Tcell=25ºC and relative efficiency and Isc gains of over 6%.

Towards 35% Efficient Concentrator Modules Based On LPI Köhler Concentrators With External Confinement Cavities

Multijunction solar cells present a certain reflectivity on its surface that lowers its light absorption. This reflectivity produces a loss in electrical efficiency and thus a loss in global energy production for CPV systems. We present here an optical design for recovering this portion of reflected light, and thus leading to a system efficiency increase. This new design is based on an external confinement cavity, an optical element able to redirect the light reflected by the cell towards its surface again. We have proven the excellent performance of these cavities integrated in CPV modules offering outstanding results: 33.2% module electrical efficiency @Tcell  =  25 °C and relative efficiency and Isc gains of over 6%

Caracterización y modelado de grandes centrales fotovoltaicas

La presente tesis doctoral propone un conjunto de ensayos de corta duración destinados a cubrir la ausencia de estándares internacionales específicamente adaptados a la tecnología y al panorama fotovoltaico actual que indiquen como realizar los procedimientos de control de calidad para comprobar que las grandes centrales fotovoltaicas ejecutadas responden a las expectativas establecidas durante la fase de proyecto. Dichos ensayos buscan, desde el punto de vista estrictamente técnico, obtener en un corto periodo de tiempo (típicamente una semana) resultados altamente repetitivos y representativos del comportamiento de la instalación bajo análisis, a la vez que minimizar al máximo la incertidumbre global, aspectos fundamentales para los procedimientos de control general de la calidad de una central. Los ensayos propuestos comprueban tanto el comportamiento general de la central, en términos de su capacidad de producción energética, como el de sus principales componentes, generadores fotovoltaicos e inversores, en términos de potencia máxima y eficiencia, respectivamente. También se aconseja una revisión de la calidad y seguridad de la instalación y de los materiales empleados en la ejecución de la central para evitar un envejecimiento prematuro de los mismos. Todos los ensayos recogidos en el texto se apoyan en la experiencia recopilada por el “Grupo de Sistemas Fotovoltaicos del Instituto de Energía Solar de la Universidad Politécnica de Madrid”, que ha estado involucrado en procedimientos de control de calidad de unas 50 centrales fotovoltaicas, con una potencia acumulada cercana a 250 MW, la mayoría de ellas instaladas en España. ABSTRACT This PhD thesis proposes a set of short-duration tests to establish quality control procedures to ensure that large photovoltaic plants fulfil the initial expectations. The motivation for this work is the lack of international standards specifically adapted to the present photovoltaic technology and its state of the art. From a strict technical point of view, these tests seek to obtain highly repetitive and representative results about the behaviour of the installation under study in a short period of time (typically one single week); and to minimize the global uncertainty. These are the two keys aspects required in quality control procedures. The proposed tests evaluate the general behaviour of the photovoltaic plants, in terms of energy production, as well as the particular behaviour of their main devices, photovoltaic arrays and inverters, in terms of maximum power and efficiency, respectively. A review of the installation quality and safety, and the employed materials in its execution to avoid premature aging is also recommended. The tests here presented are based on the experience accumulated by the “Grupo de Sistemas Fotovoltaicos del Instituto de Energia Solar de la Universidad Politecnica de Madrid”. This group has been involved in quality control procedures of about 50 photovoltaic plants, with an accumulated power close to 250 MW, most of them installed in Spain.

Análisis técnico-económico de acceso al mercado alemán EPEX SPOT

Acciona Energía dispone en Alemania de 150 MW de potencia instalada. Todos ellos de energía eólica, en un total de doce parques eólicos situados en la sur del país. El proyecto tiene por objeto comparar las distintas modalidades de venta de energía procedente de fuentes renovables que ofrece el estado alemán para la cartera de activos de Acciona Energía, llegando a una conclusión final de cuál de ellas es la más aconsejable. Para ello se ha realizado un estudio del funcionamiento y normas del mercado Epex Spot de la electricidad y de la legislación alemana correspondiente a la materia, así como un seguimiento exhaustivo de producción y otras variables de los parques eólicos para su análisis. Los cálculos y las estimaciones realizados llevaron a la conclusión, que la mejor opción era la venta directa en el mercado Epex Spot, para lo que primero habría que darse de alta como agente en dicho mercado. Aunque esta opción asuma mayores riesgos también ofrecería un aumento considerable de ingresos. ABSTRACT Acciona Energía has 150 MW of power capacity in Germany, all of them wind energy in a total of twelve installations, located in the south of the country. The main goal of the project is to compare the different ways to sell the energy which became from renewable source that German state offers for Acciona’s asset portfolio, finding the most advisable conclusion. To do so a study of standards and rules of Epex Spot electricity market and German law related to this topic has been made. In addition of an exhaustive monitoring of energy production and others wind farms variables has been analyzed. The reckoning and estimations saw the conclusion that the best option was the direct sell in Epex Spot market, in order to do that the first of all is to register as market agent. Despite this options assume bigger risks, it provide a substantial increase in income