Quantitative expansion microscopy for the characterization of the spectrin periodic skeleton of axons using fluorescence microscopy

Fluorescent nanoscopy approaches have been used to characterize the periodic organization of actin,spectrin and associated proteins in neuronal axons and dendrites. This membrane-associated periodicskeleton (MPS) is conserved across animals, suggesting it is a fundamental component of neuronalextensions. The nanoscale architecture of the arrangement (190 nm) is below the resolution limitof conventional fluorescent microscopy. Fluorescent nanoscopy, on the other hand, requires costlyequipment and special analysis routines, which remain inaccessible to most research groups. Thisreport aims to resolve this issue by using protein-retention expansion microscopy (pro-ExM) to revealthe MPS of axons. ExM uses reagents and equipment that are readily accessible in most neurobiologylaboratories. We first explore means to accurately estimate the expansion factors of protein structureswithin cells. We then describe the protocol that produces an expanded specimen that can be examinedwith any fluorescent microscopy allowing quantitative nanoscale characterization of the MPS. Wevalidate ExM results by direct comparison to stimulated emission depletion (STED) nanoscopy. Weconclude that ExM facilitates three-dimensional, multicolor and quantitative characterization of theMPS using accessible reagents and conventional fluorescent microscopes.Fil: Martínez, Gaby F.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Gazal, Nahir Guadalupe. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Quassollo Infanzon, Gonzalo Emiliano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Szalai, Alan Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias "Elizabeth Jares Erijman"; ArgentinaFil: Del Cid Pellitero, Esther. No especifíca;Fil: Durcan, Thomas M.. No especifíca;Fil: Fon, Edward A.. No especifíca;Fil: Bisbal, Mariano. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; ArgentinaFil: Stefani, Fernando Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Centro de Investigaciones en Bionanociencias "Elizabeth Jares Erijman"; ArgentinaFil: Unsain, Nicolas. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra. Universidad Nacional de Córdoba. Instituto de Investigación Médica Mercedes y Martín Ferreyra; Argentin

LegioLit: Knappable material lithotheque in the Prehistory Laboratory at the University of León, Spain

This work introduces a comparative collection located in the Prehistory Laboratory at the University of León (Spain) specialised in knappable raw materials, mainly comprising radiolarite and black chert (micro-crypto crystalline quartz), from the western Cantabrian Mountains (north of Iberian Peninsula). A standardised protocol of sample collection and data organisation was developed, which includes the use of several methodologies. First, Geographical Information Systems (GIS) for referencing lithic sources. Second, direct observation of the sample for the macroscopic characterization, both de visu and stereomicroscope. Third, petrographic microscopy for a description of main petrological, and palaeontological features, complemented with the identification of the different minerals that make up the samples by X-ray diffraction (XRD). Forth, X-Ray Fluorescence (XRF), Inductively Coupled Plasma Optical Emission Spectrometry (ICP-OES) and Thermogravimetry – Differential Scanning Calorimetry (TG-DSC) for geochemical and thermal features of the samples. Finally, the results of these analyses were entered in a database. All this information is contributing towards the creation of a physical reference collection specialised in local Palaeozoic formations (mostly from Devonian to Carboniferous) that outcrop in the western Cantabrian Mountains, a region whose potential resource base was previously not very well known. This collection would allow to compare archaeological lithic remains from different sites inside and outside the Cantabrian Mountains

Differential association between S100A4 levels and insulin resistance in prepubertal children and adult subjects with clinically severe obesity

Objectives: S100A4 has been recently identified as an adipokine associated with insulin resistance (IR) in adult subjects with obesity. However, no data about its levels in children with obesity and only a few approaches regarding its potential mechanism of action have been reported. To obtain a deeper understanding of the role of S100A4 in obesity, (a) S100A4 levels were measured in prepubertal children and adult subjects with and without obesity and studied the relationship with IR and (b) the effects of S100A4 in cultured human adipocytes and vascular smooth muscle cells (VSMCs) were determined. Methods: Sixty-five children (50 with obesity, age 9.0 ±1.1 years and 15 normal weight, age 8.4 ±0.8 years) and fifty-nine adults (43 with severe obesity, age 46 ±11 years and 16 normal weight, age 45 ±9 years) were included. Blood from children and adults and adipose tissue samples from adults were obtained and analysed. Human adipocytes and VSMC were incubated with S100A4 to evaluate their response to this adipokine. Results: Circulating S100A4 levels were increased in both children (P =.002) and adults (P <.001) with obesity compared with their normal-weight controls. In subjects with obesity, S100A4 levels were associated with homeostatic model assessment-insulin resistance (HOMA-IR) in adults (βstd =.42, P =.008) but not in children (βstd =.12, P =.356). Human adipocytes were not sensitive to S100A4, while incubation with this adipokine significantly reduced inflammatory markers in VSMC. Conclusions: Our human data demonstrate that higher S100A4 levels are a marker of IR in adults with obesity but not in prepubertal children. Furthermore, the in vitro results suggest that S100A4 might exert an anti-inflammatory effect. Further studies will be necessary to determine whether S100A4 can be a therapeutic target for obesity

A role for Oncostatin M in the impairment of glucose homeostasis in obesity

CONTEXT: Oncostatin M (OSM) plays a key role in inflammation, but its regulation and function during obesity is not fully understood. OBJECTIVE: The aim of this study was to evaluate the relationship of OSM with the inflammatory state that leads to impaired glucose homeostasis in obesity. We also assessed whether OSM immunoneutralization could revert metabolic disturbances caused by a high-fat diet (HFD) in mice. DESIGN: 28 patients with severe obesity were included and stratified into two groups: (1) glucose levels 100 mg/dL. White adipose tissue was obtained to examine OSM gene expression. Human adipocytes were used to evaluate the effect of OSM in the inflammatory response, and HFD-fed C57BL/6J mice were injected with anti-OSM antibody to evaluate its effects. RESULTS: OSM expression was elevated in subcutaneous and visceral fat from patients with obesity and hyperglycemia, and correlated with Glut4 mRNA levels, serum insulin, homeostatic model assessment of insulin resistance, and inflammatory markers. OSM inhibited adipogenesis and induced inflammation in human adipocytes. Finally, OSM receptor knockout mice had increased Glut4 mRNA levels in adipose tissue, and OSM immunoneutralization resulted in a reduction of glucose levels and Ccl2 expression in adipose tissue from HFD-fed mice. CONCLUSIONS: OSM contributes to the inflammatory state during obesity and may be involved in the development of insulin resistance

Regional development gaps in Argentina: A multidimensional approach to identify the location of policy priorities

Spatial inequalities within Latin American countries have historically attracted the interest ofacademics, policy-makers, and international agencies. This article aims to provide amultidimensional diagnosis of provincial development gaps in Argentina, in order to identifythe location of policy priorities. Therefore, we built a large database, which covers sevendevelopment dimensions, and applied multivariate analysis techniques to overcome someanalytical limitations of previous studies. Results show the stability of provincial developmentgaps between 2003 and 2013 and some heterogeneity within geographic regions. Instead,cluster analysis offers a better classification of Argentine provinces according to theirdevelopment gaps, which can help the government to prioritize the places wheredevelopment policies are strategic.Fil: Niembro, Andrés Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte; Argentina. Universidad Nacional de Río Negro; ArgentinaFil: Sarmiento, Jesica Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte; Argentina. Universidad Nacional de Río Negro; Argentin

Analysis of Modern Optical Inspection Systems for Parts Manufactured by Selective Laser Melting

Metal additive manufacturing (AM) allows obtaining functional parts with the possibility of optimizing them topologically without affecting system performance. This is of great interest for sectors such as aerospace, automotive, and medical&ndash;surgical. However, from a metrological point of view, the high requirements applied in these sectors constitute a challenge for inspecting these types of parts. Non-contact inspection has gained great relevance due to the rapid verification of AM parts. Optical measurement systems (OMSs) are being increasingly adopted for geometric dimensioning and tolerancing (GD&amp;T) verification within the context of Industry 4.0. In this paper, the suitability (advantages and limitations) of five different OMSs (based on laser triangulation, conoscopic holography, and structured light techniques) for GD&amp;T verification of parts manufactured by selective laser melting (SLM) is analyzed. For this purpose, a specific testing part was designed and SLM-manufactured in 17-4PH stainless steel. Once the part was measured by contact (obtaining the reference GD&amp;T values), it was optically measured. The scanning results allow comparing the OMSs in terms of their inspection speed as well as dimensional and geometrical accuracy. As a result, two portable systems (handheld laser triangulation and structured blue-light scanners) were identified as the most accurate optical techniques for scanning SLM parts

Establecimiento de un precio óptimo ambiental y social para el agua de riego mediante la internalización de los costes ambientales y el uso de matrices de contabilidad social. Aplicación a la Comunidad Foral de Navarra

[EN] This work develops a methodology to obtain a “social ad environmental optimal price” for water in the Comunidad Foral de Navarra (CFN) in order to maximize the internalization of the water environmental cost generated by agricultural consumption without affecting the general level of the regional economy. The economical model is built over a Geographical Information System (GIS) that allows quantifying and valuing in economical terms environmental water costs in different basins. Elements that have been considered to describe the water resource value are volume, quality and hazard to affect aquatic and bank ecosystems. Values have been estimated with Environmental Economy methods. A demand curve for water is also built and a Social Accounting Matrix (SAM) is used to watch if regional economy supports a price that internalizes total water value. Potential water prices are established ranging from prices that minimize negative economic regional impacts –social optimal priceto those that internalize totally the environmental costs of water- environmetnal optimal price.[ES] Se propone una metodología para obtener un “precio social y ambiental óptimo” que pueda aplicarse al agua de riego en la Comunidad Foral de Navarra (CFN) de modo que maximice la internalización de los costes ambientales asociados al consumo no contemplados en el actual precio sin que el nivel general de la economía regional se resienta. Para ello se elabora un modelo económico sobre un Sistema de Información Geográfica (SIG) que permite cuantificar y valorar los costes ambientales en cada una de las subcuencas de la Comunidad. Para valorar íntegramente el agua, los elementos que se tienen en cuenta son: volumen, calidad y riesgo de afectar a los ecosistemas acuáticos y de ribera. Estos valores son estimados mediante métodos de valoración económico ambiental. Se construye una curva de demanda para los regantes y un modelo de matrices de cuentas sociales (MCS). A partir de estos elementos se estudia el impacto que sobre la economía de la región tendría un incremento en el precio del agua. Se establece un rango potencial de precios del agua de riego desde el que minimiza los impactos económicos negativos –precio social óptimo- hasta el que internaliza totalmente los costes ambientales del agua- precio ambiental óptimo-.Elorrieta, J.; Castellano, E.; Martínez De Anguita, P.; Pellitero, M.; Rey, C. (2003). Environmental and social ideal price establishment for the irrigation water by means of the internalisation of the environmental costs and the use of social accounting matrix. Application to Navarre. 139-166. doi:10.7201/earn.2003.01.07SWORD13916

Laser Defocusing Effect on the Microstructure and Defects of 17-4PH Parts Additively Manufactured by SLM at a Low Energy Input

In this paper, the microstructure, defects, hardness, and tensile strength of the 17-4PH specimens manufactured additively using the selective laser melting (SLM) technique were studied. The analyzed parts (10 mm size cubic specimens and tensile specimens) were manufactured with different defocus parameter values (-1, 0, +1 mm) in order to evaluate this effect with a low power laser (38 W). The study was carried out on three different sections of each cubic specimen, one perpendicular to the laser beam or SLM manufacturing direction (transversal section), and another two parallel to the laser beam direction (longitudinal sections). The specimens microstructures were analyzed with an X-ray diffraction technique, as well as optical, scanning electron, and transmission electron microscopes. Image J software was used to characterize the defects and phase ratio. In addition, hardness and tensile tests were performed according to the corresponding standards. The results show that the amount of austenitic phase and the average grain size varied with defocusing. The percentage of defective area was less than 0.25%. The analyzed defocus distance did not affect the number and average size of the defects. Adjusting the defocusing SLM parameter is important for manufacturing parts with good mechanical properties

Modelo de evaluación y seguimiento de los Trabajos Fin de Grado (TFG) y Trabajos Fin de Máster (TFM) tutorizados en el Área de Ingeniería de los Procesos de Fabricación

Congreso Universitario de Innovación Educativa En las Enseñanzas Técnicas, CUIEET (26º. 2018. Gijón