Spinal cord metabolic signatures in models of fast- and slow-progressing SOD1G93A Amyotrophic Lateral Sclerosis

The rate of disease progression in amyotrophic lateral sclerosis (ALS) is highly variable, even between patients with the same genetic mutations. Metabolic alterations may affect disease course variability in ALS patients, but challenges in identifying the preclinical and early phases of the disease limit our understanding of molecular mechanisms underlying differences in the rate of disease progression. We examined effects of SOD1G93A on thoracic and lumbar spinal cord metabolites in two mouse ALS models with different rates of disease progression: the transgenic SOD1G93A-C57BL/6JOlaHsd (C57-G93A, slow progression) and transgenic SOD1G93A-129SvHsd (129S-G93A, fast progression) strains. Samples from three timepoints (presymptomatic, disease onset, and late stage disease) were analyzed using Gas Chromatography-Mass Spectrometry metabolomics. Tissue metabolome differences in the lumbar spinal cord were driven primarily by mouse genetic background, although larger responses were observed in metabolic trajectories after the onset of symptoms. The significantly affected lumbar spinal cord metabolites were involved in energy and lipid metabolism. In the thoracic spinal cord, metabolic differences related to genetic background, background-SOD1 genotype interactions, and longitudinal SOD1G93A effects. The largest responses in thoracic spinal cord metabolic trajectories related to SOD1G93A effects before onset of visible symptoms. More metabolites were significantly affected in the thoracic segment, which were involved in energy homeostasis, neurotransmitter synthesis and utilization, and the oxidative stress response. We find evidence that initial metabolic alterations in SOD1G93A mice confer disadvantages for maintaining neuronal viability under ALS-related stressors, with slow-progressing C57-G93A mice potentially having more favorable spinal cord bioenergetic profiles than 129S-G93A. These genetic background-associated metabolic differences together with the different early metabolic responses underscore the need to better characterize the impact of germline genetic variation on cellular responses to ALS gene mutations both before and after the onset of symptoms in order to understand their impact on disease development

Altered metabolic profiles associate with toxicity in SOD1G93A astrocyte-neuron co-cultures

Non-cell autonomous processes involving astrocytes have been shown to contribute to motor neuron degeneration in amyotrophic lateral sclerosis. Mutant superoxide dismutase 1 (SOD1G93A) expression in astrocytes is selectively toxic to motor neurons in co-culture, even when mutant protein is expressed only in astrocytes and not in neurons. To examine metabolic changes in astrocyte-spinal neuron co-cultures, we carried out metabolomic analysis by 1H NMR spectroscopy of media from astrocyte-spinal neuron co-cultures and astrocyte-only cultures. We observed increased glucose uptake with SOD1G93A expression in all co-cultures, but while co-cultures with only SOD1G93A neurons had lower extracellular lactate, those with only SOD1G93A astrocytes exhibited the reverse. Reduced branched-chain amino acid uptake and increased accumulation of 3-methyl-2-oxovalerate were observed in co-culture with only SOD1G93A neurons while glutamate was reduced in all co-cultures expressing SOD1G93A. The shifts in these coupled processes suggest a potential block in glutamate processing that may impact motor neuron survival. We also observed metabolic alterations which may relate to oxidative stress responses. Overall, the different metabolite changes observed with the two SOD1G93A cell types highlight the role of the astrocyte-motor neuron interaction in the resulting metabolic phenotype, requiring further examination of altered metabolic pathways and their impact on motor neuron survival

Conversión de un Robot Móvil de Cadenas en Vehículo Híbrido Eléctrico

Este trabajo ha sido publicado en: Actas de las XXXV Jornadas de Automática. Valencia,Comité Español de Automática CEA-IFAC , 2014. ISBN-13:78-84-697-0589-6En la remodelación del robot móvil Auriga-a, desarrollado en el año 2000, solo se ha conservado su sistema de locomoción. Además de la actualización de los sistemas informáticos y equipos de control, el principal objetivo ha sido la conversión del robot en un vehículo híbrido eléctrico. Para ello, se han incorporado baterías LiFePO4 con su correspondiente cargador y un generador monofásico. En el diseño, se han tenido en cuenta el posicionado del centro de gravedad y la ventilación de los equipos. En conjunto, la remodelación consigue reducir la masa del vehículo y mejorar su eficiencia y autonomía energética. En la actualidad, el diseño está siendo implantado en el robot real.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. CICYT DPI 2011-22443

Remote Inspection, Measurement and Handling for LHC

Personnel access to the LHC tunnel will be restricted to varying extents during the life of the machine due to radiation, cryogenic and pressure hazards. The ability to carry out visual inspection, measurement and handling activities remotely during periods when the LHC tunnel is potentially hazardous offers advantages in terms of safety, accelerator down time, and costs. The first applications identified were remote measurement of radiation levels at the start of shut-down, remote geometrical survey measurements in the collimation regions, and remote visual inspection during pressure testing and initial machine cool-down. In addition, for remote handling operations, it will be necessary to be able to transmit several real-time video images from the tunnel to the control room. The paper describes the design, development and use of a remotely controlled vehicle to demonstrate the feasibility of meeting the above requirements in the LHC tunnel. Design choices are explained along with operating experience to-date and future development plans

Influencia del calor y del aclarado sobre la germinación de Cistus laurifolius y Cistus ladanifer

The aim of this study is to determine if the increase of the temperature of the soil while a wildfire improves the germination of Cistus laurifolius and Cistus ladanifer. Laboratory experiments were carried out by heating the seeds to different temperatures and at different periods of time. On the other hand, the regeneration of both species in the field was studied in experimental plots that were subjected to prescribed burning, as well as to cutting of the aerial biomass. Cistus ladanifer regrows better than Cistus laurifolius. The answer, in terms of seedling number, is also better in the burnt plots than in the cut. The height in each case follows different patterns. The field results agree with those of the laboratory.[es] El objetivo de este estudio es determinar si el aumento de temperatura que se produce en el suelo durante los incendios favorece la germinación de las semillas de Cistus laurliolius y Cistus ladanifer. Se realizan ensayos de germinación en el laboratorio, calentando las semillas a diferentes temperaturas durante distintos tiempos. Paralelamente, se estudia la regeneración de ambas especies en condiciones de campo, en parcelas experimentales sometidas a quema controlada y corta de la biomasa aérea. Cistus ladanifer se regenera mejor que Cistus laurifolius, en términos de número de plántulas, y mejor en las parcelas quemadas que en las cortadas. La altura en cada caso sigue patrones diferentes. Los resultados de campo concuerdan con los de laboratorio. [fr] L'objectif de cette étude consiste à déterminer l'augmentation de la température dans le sol pendant les incendies favorise la germination des semences de Cistus laurifolius et Cistus ladanifer. On a réalisé des essais de germination au laboratoire, en faisant chauffer les semences à plusieurs températures pendant des temps différents. En même temps on a étudié la régénération de ces deux espèces sur le terrain, dans des parcelles expérimentales soumises à un brûlage contrôlé et court de la biomasse aérienne. Cistus ladanifer se régénère mieux que Cistus laurifolius, en ce qui concerne le nombre de plantules, et mieux dans les parcelles brûlées que dans celles coupées. La hauteur a suivi des modèles différents. Les résultats de terrain concordent avec ceux obtenus au laboratoire

The 14q32 maternally imprinted locus is a major source of longitudinally stable circulating microRNAs as measured by small RNA sequencing

Understanding the normal temporal variation of serum molecules is a critical factor for identifying useful candidate biomarkers for the diagnosis and prognosis of chronic disease. Using small RNA sequencing in a longitudinal study of 66 women with no history of cancer, we determined the distribution and dynamics (via intraclass correlation coefficients, ICCs) of the miRNA profile over 3 time points sampled across 2–5 years in the course of the screening trial, UKCTOCS. We were able to define a subset of longitudinally stable miRNAs (ICC >0.75) that were individually discriminating of women who had no cancer over the study period. These miRNAs were dominated by those originating from the C14MC cluster that is subject to maternal imprinting. This assessment was not significantly affected by common confounders such as age, BMI or time to centrifugation nor alternative methods to data normalisation. Our analysis provides important benchmark data supporting the development of miRNA biomarkers for the impact of life-course exposure as well as diagnosis and prognostication of chronic disease

ESCargo: a regulatable fluorescent secretory cargo for diverse model organisms

© The Author(s), 2020. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Casler, J. C., Zajac, A. L., Valbuena, F. M., Sparvoli, D., Jeyifous, O., Turkewitz, A. P., Horne-Badovinac, S., Green, W. N., & Glick, B. S. ESCargo: a regulatable fluorescent secretory cargo for diverse model organisms. Molecular Biology of the Cell, (2020): mbcE20090591, doi:10.1091/mbc.E20-09-0591.Membrane traffic can be studied by imaging a cargo protein as it transits the secretory pathway. The best tools for this purpose initially block export of the secretory cargo from the endoplasmic reticulum (ER), and then release the block to generate a cargo wave. However, previously developed regulatable secretory cargoes are often tricky to use or specific for a single model organism. To overcome these hurdles for budding yeast, we recently optimized an artificial fluorescent secretory protein that exits the ER with the aid of the Erv29 cargo receptor, which is homologous to mammalian Surf4. The fluorescentsecretory protein forms aggregates in the ER lumen and can be rapidly disaggregated by addition of a ligand to generate a nearly synchronized cargo wave. Here we term this regulatable secretory proteinESCargo (Erv29/Surf4-dependent Secretory Cargo) and demonstrate its utility not only in yeast cells, but also in cultured mammalian cells, Drosophila cells, and the ciliate Tetrahymena thermophila. Kinetic studies indicate that rapid export from the ER requires recognition by Erv29/Surf4. By choosing an appropriate ER signal sequence and expression vector, this simple technology can likely be used withmany model organisms.This work was supported by NIH grant R01 GM104010 to BSG, by NIH grant R01 GM105783 to APT, by NIH grant R01 GM136961 and American Cancer Society grant RSG-14-176 to SHB, and by NIH grant R01 DA044760 to WNG. JCC was supported by NIH training grant T32 GM007183. AZ was supported by American Heart Association fellowship 16POST2726018 and American Cancer Society fellowship 132123-PF-18-025-01-CSM. Thanks for assistance with fluorescence microscopy to Vytas Bindokas and Christine Labno at the Integrated Microscopy Core Facility, which is supported by the NIH-funded Cancer Center Support Grant P30 CA014599. The pUASt-ManII-eGFP plasmid was a gift from Bing Ye, and the Ubi-Gal4 plasmid was a gift from Rick Fehon.2020-12-2

Fructose transporter Glut5 expression in clear renal cell carcinoma

[Abstract] Renal cell carcinomas (RCC) can be subclassified for general purposes into clear cell, papillary cell, chromophobe cell carcinomas and oncocytomas. Other tumours such as collecting duct, medullary, mucinous tubular and spindle cell and associated with Xp 11.2 translocations/TFE 3 gene fusion, are much less common. There is also a residual group of unclassified cases. Previous studies have shown that RCC has high glycolytic rates, and expresses GLUT transporters, but no distinction has been made among the different subtypes of renal cell tumours and their grades of malignancy. In clear renal cell carcinoma (cRCC) glycogen levels increase, glycolysis is activated and gluconeogenesis is reduced. The clear cell subtype of RCC is characterized histologically by a distinctive pale, glassy cytoplasm and this appearance of cRCC is due to abnormalities in carbohydrate and lipid metabolism, and this abnormality results in glycogen and sterol storage. Several isoforms of glucose carriers (GLUTs) have been identified. We show here in a panel of 80 cRCC samples a significant correlation between isoform 5 (GLUT5) and many pathological parameters such as grade of differentiation, pelvis invasion and breaking capsule. GLUT5 expression also appears to associate more strongly with the clear cell RCC subtype. These data suggest a role for the GLUT5 isoform in fructose uptake that takes place in cRCC cells and which subsequently leads to the malignant RCC progression