Input-Output Based Economic Impact Evaluation System for Small City Development: A Case Study on Saemangeum's Flux City Design

The paper aims to develop a quasi-dynamic interregional input-output model for evaluating the macro-economic impacts of small city development. The features of the model are summarized as follows: (1) the consumption expenditure of households is regarded as an endogenous variable, (2) the technological change is determined by the change of industrial Location Quotient caused by firm's investment activities. (3) a strong feedback function between the city design and the economic analysis is provided. For checking the performance of the model, Saemangeum's Flux City Design Plan is used as the simulation target in our paper.

Assessment of CTF boiling transition and critical heat flux modeling capabilities using the OECD/NRC BFBT and PSBT benchmark databases

The need to refine models for best-estimate calculations, based on good-quality experimental data, has been expressed in many recent meetings in the field of nuclear applications. The modeling needs arising in this respect should not be limited to the currently available macroscopic methods but should be extended to next-generation analysis techniques that focus on more microscopic processes. One of the most valuable databases identified for the thermalhydraulics modeling was developed by the Nuclear Power Engineering Corporation (NUPEC), Japan. From 1987 to 1995, NUPEC performed steady-state and transient critical power and departure from nucleate boiling (DNB) test series based on the equivalent full-size mock-ups. Considering the reliability not only of the measured data, but also other relevant parameters such as the system pressure, inlet sub-cooling and rod surface temperature, these test series supplied the first substantial database for the development of truly mechanistic and consistent models for boiling transition and critical heat flux. Over the last few years the Pennsylvania State University (PSU) under the sponsorship of the U.S. Nuclear Regulatory Commission (NRC) has prepared, organized, conducted and summarized the OECD/NRC Full-size Fine-mesh Bundle Tests (BFBT) Benchmark. The international benchmark activities have been conducted in cooperation with the Nuclear Energy Agency/Organization for Economic Co-operation and Development (NEA/OECD) and Japan Nuclear Energy Safety (JNES) organization, Japan. Consequently, the JNES has made available the Boiling Water Reactor (BWR) NUPEC database for the purposes of the benchmark. Based on the success of the OECD/NRC BFBT benchmark the JNES has decided to release also the data based on the NUPEC Pressurized Water Reactor (PWR) subchannel and bundle tests for another follow-up international benchmark entitled OECD/NRC PWR Subchannel and Bundle Tests (PSBT) benchmark. This paper presents an application of the joint Penn State University/Technical University of Madrid (UPM) version of the well-known subchannel code COBRA-TF, namely CTF, to the critical power and departure from nucleate boiling (DNB) exercises of the OECD/NRC BFBT and PSBT benchmarks

Effects of diet and exercise on metabolic syndrome and the underlying factors = Efectos de la dieta y el ejercicio sobre el síndrome metabólico y sus factores

Introducción. El número de personas que padecen síndrome metabólico ha incrementado a nivel mundial durante las últimas dos décadas. Existen numerosos estudios que tratan de comparar prevalencias según los diferentes criterios y estimaciones del riesgo metabólico. De ellos se puede concluir que el principal hallazgo ha sido recalcar la necesidad de una definición estándar universal. A pesar de estas discrepancias no hay lugar a duda sobre el problema de salud pública que esto conlleva. Se necesitan medidas y estrategias urgentes para prevenir y controlar esta emergente epidemia global y para ello se debe prestar especial atención a los cambios en el estilo de vida, fundamentalmente dieta y ejercicio. A pesar de todo, existe a día de hoy una importante controversia sobre el tipo de ejercicio más efectivo y su combinación con la dieta para conseguir mejoras en la salud. Objetivos. Estudiar los índices de riesgo metabólico empleados en la literatura científica y las terapias basadas en dieta y ejercicio para el tratamiento de los factores del síndrome metabólico en adultos con sobrepeso. Diseño de investigación. Los datos empleados en el análisis de esta tesis son, primeramente un estudio piloto, y posteriormente parte del estudio “Programas de Nutrición y Actividad Física para el tratamiento de la obesidad” (PRONAF). El estudio PRONAF es un proyecto consistente en un estudio clínico sobre programas de nutrición y actividad física para el sobrepeso y la obesidad, desarrollado en España durante varios años de intervenciones. Fue diseñado, en parte, para tratar de comparar protocolos de entrenamiento de resistencia, cargas y combinado en igualdad de volumen e intensidad, con el objetivo de evaluar su impacto en los factores de riesgo y la prevalencia del síndrome metabólico en personas con sobrepeso y obesidad. El diseño experimental es un control aleatorio y el protocolo incluye 3 modos de ejercicio (entrenamiento de resistencia, con cargas y combinado) y restricción dietética sobre diversas variables determinantes del estado de salud. Las principales variables para la investigación que comprende esta tesis fueron: actividad física habitual, marcadores de grasa corporal, niveles de insulina, glucosa, triglicéridos, colesterol total, colesterol HDL, colesterol LDL, presión arterial y parámetros relacionados con el ejercicio. Conclusiones. A) Los índices de riesgo metabólico estudiados presentan resultados contradictorios en relación al riesgo metabólico en un individuo, dependiendo de los métodos matemáticos empleados para el cálculo y de las variables introducidas, tanto en mujeres sanas como en adultos en sobrepeso. B) El protocolo de entrenamiento combinado (de cargas y de resistencia) junto con la dieta equilibrada propuesto en este estudio fue la mejor estrategia para la mejora del riesgo de síndrome metabólico en adultos con sobrepeso. C) Los protocolos de entrenamiento supervisado de resistencia, con cargas y combinado junto con la restricción nutricional, no obtuvieron mejoras sobre el perfil lipídico, más allá de los cambios conseguidos con el protocolo de dieta y recomendaciones generales de actividad física habitual en clínica, en adultos con sobrepeso. Background. Over the past two decades, a striking increase in the number of people with the MetS worldwide has taken place. Many studies compare prevalences using different criteria and metabolic risk estimation formulas, and perhaps their main achievement is to reinforce the need for a standardized international definition. Although these discrepancies, there is no doubt it is a public health problem. There is urgent need for strategies to prevent and manage the emerging global epidemic, special consideration should be given to behavioral and lifestyle, mainly diet and exercise. However, there is still controversy about the most effective type of exercise and diet combination to achieve improvements. Objectives. To study the metabolic risk scores used in the literature and the diet and exercise therapies for the treatment of the MetS factors in overweight adults. Research design. The data used in the analysis was collected firstly in a pilot study and lately, as a part of the “Programas de Nutrición y Actividad física para el tratamiento de la obesidad” study (PRONAF). The PRONAF Study is a clinical research project in nutrition and physical activity programs for overweight and obesity, carried out in Spain (2008-2011). Was designed, in part, to attempt to match the volume and intensity of endurance, strength and combined training protocols in order to evaluate their impact on risk factors and MetS prevalence in overweight and obese people. The design and protocol included three exercise modes (endurance, strength and combined training) and diet restriction, in a randomized controlled trial concerning diverse health status variables. The main variables under investigation were habitual physical activity, markers of body fat, fasting serum levels of insulin, glucose, triglycerides, total, LDL and HDL cholesterol, blood pressure and diet and exercise parameters. Main outcomes. A) The metabolic risk scores studied presented contradictory results in relation to the metabolic risk of an individual, depending on the mathematical method used and the variables included, both in healthy women and overweight adults. B) The protocol proposed for combination of strength and endurance training combined with a balance diet was the optimal strategy for the improvement of MetS risk in overweight adults. C) The intervention program of endurance, strength or combined supervised training protocol with diet restriction did not achieved further improvements in lipid profile than a habitual clinical practice protocol including dietary advice and standard physical activity recommendations, in overweight adults.

Finite Element Analysis Model of a Contactless Transformer for Battery Chargers in Electric Vehicles

A contactless transformer model is proposed in this paper using Finite Element Analysis (FEA). This model can be used to simulate Inductive Coupling Power Transfer (ICPT) systems with good accuracy of the transformer and reduce the fabrication time of these systems. The model not only takes into account the geometry of the windings but also the frequency effects in them. As the transformer does not have a magnetic core, it is complicated to model because the flux is expanded in the area around the windings. In order to obtain a very accurate model, it is necessary to use a 2D/3D field solver.

Neutronic analysis of the bi-spectral moderator such as that proposed for ESS

This paper reports on design studies concerning a moderator concept which aims to maximize the time averaged flux. The idea is to provide neutron spectra showing two clear maxima, with peaks at View the MathML source and View the MathML source arising from leakage from both cryogenic and thermal moderators. Such a concept known as a bi-spectral moderator (Mezei, 2006 [1]) while proven on a reactor source has only been examined for the ESS 2003 proposal. Filges et al. (2003 [2]), which featured a different design than the current ESS. This paper thus reports on a baseline design for such a moderator concept and shows that it can provide substantial gains in count rates for those applications not requiring high resolution in time-of-flight.

Analysis of tape tether survival in LEO against orbital debris

The low earth orbit (LEO) environment contains a large number of artificial debris, of which a significant portion is due to dead satellites and fragments of satellites resulted from explosions and in-orbit collisions. Deorbiting defunct satellites at the end of their life can be achieved by a successful operation of an Electrodynamic Tether (EDT) system. The effectiveness of an EDT greatly depends on the survivability of the tether, which can become debris itself if cut by debris particles; a tether can be completely cut by debris having some minimal diameter. The objective of this paper is to develop an accurate model using power laws for debris-size ranges, in both ORDEM2000 and MASTER2009 debris flux models, to calculate tape tether survivability. The analytical model, which depends on tape dimensions (width, thickness) and orbital parameters (inclinations, altitudes) is then verified with fully numerical results to compare for different orbit inclinations, altitudes and tape width for both ORDEM2000 and MASTER2009 flux data.

Validation of the burn-up code EVOLCODE 2.0 with PWR experimental data and with a Sensitivity/Uncertainty analysis

A validation of the burn-up simulation system EVOLCODE 2.0 is presented here, involving the experimental measurement of U and Pu isotopes and some fission fragments production ratios after a burn-up of around 30 GWd/tU in a Pressurized Light Water Reactor (PWR). This work provides an in-depth analysis of the validation results, including the possible sources of the uncertainties. An uncertainty analysis based on the sensitivity methodology has been also performed, providing the uncertainties in the isotopic content propagated from the cross sections uncertainties. An improvement of the classical Sensitivity/ Uncertainty (S/U) model has been developed to take into account the implicit dependence of the neutron flux normalization, that is, the effect of the constant power of the reactor. The improved S/U methodology, neglected in this kind of studies, has proven to be an important contribution to the explanation of some simulation-experiment discrepancies for which, in general, the cross section uncertainties are, for the most relevant actinides, an important contributor to the simulation uncertainties, of the same order of magnitude and sometimes even larger than the experimental uncertainties and the experiment- simulation differences. Additionally, some hints for the improvement of the JEFF3.1.1 fission yield library and for the correction of some errata in the experimental data are presented.

High power beam dump project for the accelerator prototype LIPAc: Cooling design and analysis

En el campo de la fusión nuclear y desarrollándose en paralelo a ITER (International Thermonuclear Experimental Reactor), el proyecto IFMIF (International Fusion Material Irradiation Facility) se enmarca dentro de las actividades complementarias encaminadas a solucionar las barreras tecnológicas que aún plantea la fusión. En concreto IFMIF es una instalación de irradiación cuya misión es caracterizar materiales resistentes a condiciones extremas como las esperadas en los futuros reactores de fusión como DEMO (DEMOnstration power plant). Consiste de dos aceleradores de deuterones que proporcionan un haz de 125 mA y 40 MeV cada uno, que al colisionar con un blanco de litio producen un flujo neutrónico intenso (1017 neutrones/s) con un espectro similar al de los neutrones de fusión [1], [2]. Dicho flujo neutrónico es empleado para irradiar los diferentes materiales candidatos a ser empleados en reactores de fusión, y las muestras son posteriormente examinadas en la llamada instalación de post-irradiación. Como primer paso en tan ambicioso proyecto, una fase de validación y diseño llamada IFMIFEVEDA (Engineering Validation and Engineering Design Activities) se encuentra actualmente en desarrollo. Una de las actividades contempladas en esta fase es la construcción y operación de una acelarador prototipo llamado LIPAc (Linear IFMIF Prototype Accelerator). Se trata de un acelerador de deuterones de alta intensidad idéntico a la parte de baja energía de los aceleradores de IFMIF. Los componentes del LIPAc, que será instalado en Japón, son suministrados por diferentes países europeos. El acelerador proporcionará un haz continuo de deuterones de 9 MeV con una potencia de 1.125 MW que tras ser caracterizado con diversos instrumentos deberá pararse de forma segura. Para ello se requiere un sistema denominado bloque de parada (Beam Dump en inglés) que absorba la energía del haz y la transfiera a un sumidero de calor. España tiene el compromiso de suministrar este componente y CIEMAT (Centro de Investigaciones Energéticas Medioambientales y Tecnológicas) es responsable de dicha tarea. La pieza central del bloque de parada, donde se para el haz de iones, es un cono de cobre con un ángulo de 3.5o, 2.5 m de longitud y 5 mm de espesor. Dicha pieza está refrigerada por agua que fluye en su superficie externa por el canal que se forma entre el cono de cobre y otra pieza concéntrica con éste. Este es el marco en que se desarrolla la presente tesis, cuyo objeto es el diseño del sistema de refrigeración del bloque de parada del LIPAc. El diseño se ha realizado utilizando un modelo simplificado unidimensional. Se han obtenido los parámetros del agua (presión, caudal, pérdida de carga) y la geometría requerida en el canal de refrigeración (anchura, rugosidad) para garantizar la correcta refrigeración del bloque de parada. Se ha comprobado que el diseño permite variaciones del haz respecto a la situación nominal siendo el flujo crítico calorífico al menos 2 veces superior al nominal. Se han realizado asimismo simulaciones fluidodinámicas 3D con ANSYS-CFX en aquellas zonas del canal de refrigeración que lo requieren. El bloque de parada se activará como consecuencia de la interacción del haz de partículas lo que impide cualquier cambio o reparación una vez comenzada la operación del acelerador. Por ello el diseño ha de ser muy robusto y todas las hipótesis utilizadas en la realización de éste deben ser cuidadosamente comprobadas. Gran parte del esfuerzo de la tesis se centra en la estimación del coeficiente de transferencia de calor que es determinante en los resultados obtenidos, y que se emplea además como condición de contorno en los cálculos mecánicos. Para ello por un lado se han buscado correlaciones cuyo rango de aplicabilidad sea adecuado para las condiciones del bloque de parada (canal anular, diferencias de temperatura agua-pared de decenas de grados). En un segundo paso se han comparado los coeficientes de película obtenidos a partir de la correlación seleccionada (Petukhov-Gnielinski) con los que se deducen de simulaciones fluidodinámicas, obteniendo resultados satisfactorios. Por último se ha realizado una validación experimental utilizando un prototipo y un circuito hidráulico que proporciona un flujo de agua con los parámetros requeridos en el bloque de parada. Tras varios intentos y mejoras en el experimento se han obtenido los coeficientes de película para distintos caudales y potencias de calentamiento. Teniendo en cuenta la incertidumbre de las medidas, los valores experimentales concuerdan razonablemente bien (en el rango de 15%) con los deducidos de las correlaciones. Por motivos radiológicos es necesario controlar la calidad del agua de refrigeración y minimizar la corrosión del cobre. Tras un estudio bibliográfico se identificaron los parámetros del agua más adecuados (conductividad, pH y concentración de oxígeno disuelto). Como parte de la tesis se ha realizado asimismo un estudio de la corrosión del circuito de refrigeración del bloque de parada con el doble fin de determinar si puede poner en riesgo la integridad del componente, y de obtener una estimación de la velocidad de corrosión para dimensionar el sistema de purificación del agua. Se ha utilizado el código TRACT (TRansport and ACTivation code) adaptándalo al caso del bloque de parada, para lo cual se trabajó con el responsable (Panos Karditsas) del código en Culham (UKAEA). Los resultados confirman que la corrosión del cobre en las condiciones seleccionadas no supone un problema. La Tesis se encuentra estructurada de la siguiente manera: En el primer capítulo se realiza una introducción de los proyectos IFMIF y LIPAc dentro de los cuales se enmarca esta Tesis. Además se describe el bloque de parada, siendo el diseño del sistema de rerigeración de éste el principal objetivo de la Tesis. En el segundo y tercer capítulo se realiza un resumen de la base teórica así como de las diferentes herramientas empleadas en el diseño del sistema de refrigeración. El capítulo cuarto presenta los resultados del relativos al sistema de refrigeración. Tanto los obtenidos del estudio unidimensional, como los obtenidos de las simulaciones fluidodinámicas 3D mediante el empleo del código ANSYS-CFX. En el quinto capítulo se presentan los resultados referentes al análisis de corrosión del circuito de refrigeración del bloque de parada. El capítulo seis se centra en la descripción del montaje experimental para la obtención de los valores de pérdida de carga y coeficiente de transferencia del calor. Asimismo se presentan los resultados obtenidos en dichos experimentos. Finalmente encontramos un capítulo de apéndices en el que se describen una serie de experimentos llevados a cabo como pasos intermedios en la obtención del resultado experimental del coeficiente de película. También se presenta el código informático empleado para el análisis unidimensional del sistema de refrigeración del bloque de parada llamado CHICA (Cooling and Heating Interaction and Corrosion Analysis). ABSTRACT In the nuclear fusion field running in parallel to ITER (International Thermonuclear Experimental Reactor) as one of the complementary activities headed towards solving the technological barriers, IFMIF (International Fusion Material Irradiation Facility) project aims to provide an irradiation facility to qualify advanced materials resistant to extreme conditions like the ones expected in future fusion reactors like DEMO (DEMOnstration Power Plant). IFMIF consists of two constant wave deuteron accelerators delivering a 125 mA and 40 MeV beam each that will collide on a lithium target producing an intense neutron fluence (1017 neutrons/s) with a similar spectra to that of fusion neutrons [1], [2]. This neutron flux is employed to irradiate the different material candidates to be employed in the future fusion reactors, and the samples examined after irradiation at the so called post-irradiative facilities. As a first step in such an ambitious project, an engineering validation and engineering design activity phase called IFMIF-EVEDA (Engineering Validation and Engineering Design Activities) is presently going on. One of the activities consists on the construction and operation of an accelerator prototype named LIPAc (Linear IFMIF Prototype Accelerator). It is a high intensity deuteron accelerator identical to the low energy part of the IFMIF accelerators. The LIPAc components, which will be installed in Japan, are delivered by different european countries. The accelerator supplies a 9 MeV constant wave beam of deuterons with a power of 1.125 MW, which after being characterized by different instruments has to be stopped safely. For such task a beam dump to absorb the beam energy and take it to a heat sink is needed. Spain has the compromise of delivering such device and CIEMAT (Centro de Investigaciones Energéticas Medioambientales y Tecnológicas) is responsible for such task. The central piece of the beam dump, where the ion beam is stopped, is a copper cone with an angle of 3.5o, 2.5 m long and 5 mm width. This part is cooled by water flowing on its external surface through the channel formed between the copper cone and a concentric piece with the latter. The thesis is developed in this realm, and its objective is designing the LIPAc beam dump cooling system. The design has been performed employing a simplified one dimensional model. The water parameters (pressure, flow, pressure loss) and the required annular channel geometry (width, rugoisty) have been obtained guaranteeing the correct cooling of the beam dump. It has been checked that the cooling design allows variations of the the beam with respect to the nominal position, being the CHF (Critical Heat Flux) at least twice times higher than the nominal deposited heat flux. 3D fluid dynamic simulations employing ANSYS-CFX code in the beam dump cooling channel sections which require a more thorough study have also been performed. The beam dump will activateasaconsequenceofthe deuteron beam interaction, making impossible any change or maintenance task once the accelerator operation has started. Hence the design has to be very robust and all the hypotheses employed in the design mustbecarefully checked. Most of the work in the thesis is concentrated in estimating the heat transfer coefficient which is decisive in the obtained results, and is also employed as boundary condition in the mechanical analysis. For such task, correlations which applicability range is the adequate for the beam dump conditions (annular channel, water-surface temperature differences of tens of degrees) have been compiled. In a second step the heat transfer coefficients obtained from the selected correlation (Petukhov- Gnielinski) have been compared with the ones deduced from the 3D fluid dynamic simulations, obtaining satisfactory results. Finally an experimental validation has been performed employing a prototype and a hydraulic circuit that supplies a flow with the requested parameters in the beam dump. After several tries and improvements in the experiment, the heat transfer coefficients for different flows and heating powers have been obtained. Considering the uncertainty in the measurements the experimental values agree reasonably well (in the order of 15%) with the ones obtained from the correlations. Due to radiological reasons the quality of the cooling water must be controlled, hence minimizing the copper corrosion. After performing a bibligraphic study the most adequate water parameters were identified (conductivity, pH and dissolved oxygen concentration). As part of this thesis a corrosion study of the beam dump cooling circuit has been performed with the double aim of determining if corrosion can pose a risk for the copper beam dump , and obtaining an estimation of the corrosion velocitytodimension the water purification system. TRACT code(TRansport and ACTivation) has been employed for such study adapting the code for the beam dump case. For such study a collaboration with the code responsible (Panos Karditsas) at Culham (UKAEA) was established. The work developed in this thesis has supposed the publication of three articles in JCR journals (”Journal of Nuclear Materials” y ”Fusion Engineering and Design”), as well as presentations in more than four conferences and relevant meetings.

Mahogany (mg) stimulates feeding and increases basal metabolic rate independent of its suppression of agouti

The mahogany (mg) locus originally was identified as a recessive suppressor of agouti, a locus encoding a skin peptide that modifies coat color by antagonizing the melanocyte-stimulating hormone receptor or MC1-R. Certain dominant alleles of agouti cause an obesity syndrome when ectopic expression of the peptide aberrantly antagonizes the MC4-R, a related melanocyte-stimulating hormone receptor expressed in hypothalamic circuitry and involved in the regulation of feeding behavior and metabolism. Recent work has demonstrated that mg, when homozygous, blocks not only the ability of agouti to induce a yellow coat color when expressed in the skin of the lethal yellow mouse (AY), but also the obesity resulting from ectopic expression of agouti in the brain. Detailed analysis of mg/mg AY/a animals, presented here, demonstrates that mg/mg blocks the obesity, hyperinsulinemia, and increased linear growth induced by ectopic expression of the agouti peptide. Remarkably, however, mg/mg did not reduce hyperphagia in the AY/a mouse. Furthermore, mg/mg induced hyperphagia and an increase in basal metabolic rate in the C57BL/6J mouse in the absence of AY. Consequently, although mahogany is broadly required for agouti peptide action, it also appears to be involved in the control of metabolic rate and feeding behavior independent of its suppression of agouti.

Independent regulation of JNK/p38 mitogen-activated protein kinases by metabolic oxidative stress in the liver

The stress-activated protein kinases JNK and p38 mediate increased gene expression and are activated by environmental stresses and proinflammatory cytokines. Using an in vivo model in which oxidative stress is generated in the liver by intracellular metabolism, rapid protein–DNA complex formation on stress-activated AP-1 target genes was observed. Analysis of the induced binding complexes indicates that c-fos, c-jun, and ATF-2 were present, but also two additional jun family members, JunB and JunD. Activation of JNK precedes increased AP-1 DNA binding. Furthermore, JunB was shown to be a substrate for JNK, and phosphorylation requires the N-terminal activation domain. Unexpectedly, p38 activity was found to be constitutively active in the liver and was down-regulated through selective dephosphorylation following oxidative stress. One potential mechanism for p38 dephosphorylation is the rapid stress-induced activation of the phosphatase MKP-1, which has high affinity for phosphorylated p38 as a substrate. These data demonstrate that there are mechanisms for independent regulation of the JNK and p38 mitogen-activated protein kinase signal transduction pathways after metabolic oxidative stress in the liver.

The Escherichia coli MG1655 in silico metabolic genotype: Its definition, characteristics, and capabilities

The Escherichia coli MG1655 genome has been completely sequenced. The annotated sequence, biochemical information, and other information were used to reconstruct the E. coli metabolic map. The stoichiometric coefficients for each metabolic enzyme in the E. coli metabolic map were assembled to construct a genome-specific stoichiometric matrix. The E. coli stoichiometric matrix was used to define the system's characteristics and the capabilities of E. coli metabolism. The effects of gene deletions in the central metabolic pathways on the ability of the in silico metabolic network to support growth were assessed, and the in silico predictions were compared with experimental observations. It was shown that based on stoichiometric and capacity constraints the in silico analysis was able to qualitatively predict the growth potential of mutant strains in 86% of the cases examined. Herein, it is demonstrated that the synthesis of in silico metabolic genotypes based on genomic, biochemical, and strain-specific information is possible, and that systems analysis methods are available to analyze and interpret the metabolic phenotype.

Aging accentuates and bone marrow transplantation ameliorates metabolic defects in Fabry disease mice

Fabry disease is an X-linked metabolic disorder caused by a deficiency of α-galactosidase A (α-Gal A). The enzyme defect leads to the systemic accumulation of glycosphingolipids with α-galactosyl moieties consisting predominantly of globotriaosylceramide (Gb3). In patients with this disorder, glycolipid deposition in endothelial cells leads to renal failure and cardiac and cerebrovascular disease. Recently, we generated α-Gal A gene knockout mouse lines and described the phenotype of 10-week-old mice. In the present study, we characterize the progression of the disease with aging and explore the effects of bone marrow transplantation (BMT) on the phenotype. Histopathological analysis of α-Gal A −/0 mice revealed subclinical lesions in the Kupffer cells in the liver and macrophages in the skin with no gross lesions in the endothelial cells. Gb3 accumulation and pathological lesions in the affected organs increased with age. Treatment with BMT from the wild-type mice resulted in the clearance of accumulated Gb3 in the liver, spleen, and heart with concomitant elevation of α-Gal A activity. These findings suggest that BMT may have a potential role in the management of patients with Fabry disease.

Directed evolution of a (βα)8-barrel enzyme to catalyze related reactions in two different metabolic pathways

Enzymes participating in different metabolic pathways often have similar catalytic mechanisms and structures, suggesting their evolution from a common ancestral precursor enzyme. We sought to create a precursor-like enzyme for N′-[(5′-phosphoribosyl)formimino]-5-aminoimidazole-4-carboxamide ribonucleotide (ProFAR) isomerase (HisA; EC 5.3.1.16) and phosphoribosylanthranilate (PRA) isomerase (TrpF; EC 5.3.1.24), which catalyze similar reactions in the biosynthesis of the amino acids histidine and tryptophan and have a similar (βα)8-barrel structure. Using random mutagenesis and selection, we generated several HisA variants that catalyze the TrpF reaction both in vivo and in vitro, and one of these variants retained significant HisA activity. A more detailed analysis revealed that a single amino acid exchange could establish TrpF activity on the HisA scaffold. These findings suggest that HisA and TrpF may have evolved from an ancestral enzyme of broader substrate specificity and underscore that (βα)8-barrel enzymes are very suitable for the design of new catalytic activities.

Delineating developmental and metabolic pathways in vivo by expression profiling using the RIKEN set of 18,816 full-length enriched mouse cDNA arrays

We have systematically characterized gene expression patterns in 49 adult and embryonic mouse tissues by using cDNA microarrays with 18,816 mouse cDNAs. Cluster analysis defined sets of genes that were expressed ubiquitously or in similar groups of tissues such as digestive organs and muscle. Clustering of expression profiles was observed in embryonic brain, postnatal cerebellum, and adult olfactory bulb, reflecting similarities in neurogenesis and remodeling. Finally, clustering genes coding for known enzymes into 78 metabolic pathways revealed a surprising coordination of expression within each pathway among different tissues. On the other hand, a more detailed examination of glycolysis revealed tissue-specific differences in profiles of key regulatory enzymes. Thus, by surveying global gene expression by using microarrays with a large number of elements, we provide insights into the commonality and diversity of pathways responsible for the development and maintenance of the mammalian body plan.