Role of microRNAs in Jaagsiekte sheep retrovirus infection

Ovine pulmonary adenocarcinoma (OPA) is a lung cancer that affects sheep, caused by jaagsiekte sheep retrovirus (JSRV). OPA is present in most sheeprearing countries of the world, but, at present, there are no reliable early-stage tests to diagnose the disease, and OPA continues to pose an animal welfare threat and cause substantial economic losses. In addition, OPA is a valuable animal model to study early oncogenic events in human lung cancer. Specifically, OPA and some types of human lung cancer present similarities in activated signalling pathways (Ras-MEK-ERK1/2 and PI3K-AKT-mTOR) and their association with type II pneumocytes. Nevertheless, study of the molecular pathogenesis of OPA has been hindered due to the lack of a permissive cell line for JSRV replication. JSRV encodes an unusual envelope protein (Env) which is actively oncogenic and sufficient to drive transformation in vivo and in vitro. Despite the lack of a permissive cell line, early oncogenic events induced by JSRV can be studied by transfection of cell lines with plasmids encoding JSRV Env. MicroRNAs (miRNAs) are short non-coding RNAs that regulate gene expression and essential cell processes such as cell proliferation and apoptosis. miRNAs are being extensively studied as biomarkers of several diseases, including cancer. The aims of this project were to investigate the role of miRNAs in the early oncogenic events induced by JSRV and to investigate their potential as OPA biomarkers. miRNA expression levels were investigated using small RNA sequencing in lung tissue from cases of experimentally induced OPA. No evidence of JSRV-encoded miRNAs was found, but levels of 40 miRNAs were found differentially expressed between affected and control sheep. Of those, upregulation of nine microRNAs (miR-135b, miR-182, miR-183, miR-21, miR-200b, miR-205, miR-31, miR-503 and miR-96) was confirmed by RT-qPCR in experimental and natural cases of OPA, suggesting that increased levels of these miRNAs were characteristic of OPA affected lung tissue. To investigate miRNAs as potential biomarkers, miRNA expression was measured in serum and bronchoalveolar lavage fluid (BALF) macrophages of OPA affected sheep. small RNA sequencing revealed 74 microRNAs and 85 miRNAs differentially expressed in serum and BALF macrophages, respectively. Interestingly, BALF macrophage microRNA expression was found to resemble more closely that of OPA affected sheep lungs. In addition, miRNA expression levels varied at different stages of the disease and no miRNAs were found to be consistently dysregulated in serum of OPA affected animals. Discordances in miRNA signatures in lung tissue and serum are not entirely unexpected. Lung tissue miRNAs might represent the tumour microenvironment and localised response to it, whereas miRNAs in serum may represent the global state of the animal, and tumour miRNAs might be released into circulation at low levels, making them difficult to detect. Expression of the nine upregulated miRNAs was then investigated in in vitro models to study their involvement in transformation. Lentiviral vectors encoding green fluorescent protein (GFP) or a GFP-2A-Env fusion protein were produced and used to transduce cell lines. Transformation was verified by immunocytochemical detection of the transformation markers P-Akt and PERK1/2. Nevertheless, miRNA expression levels in culture did not resemble those observed in lung tissue of OPA-affected sheep. These differences might be due to species variation, upregulation of miRNAs late in the transformation process, or involvement of other cell types in tissue besides the transformed cells. To study these questions further, JSRV and the GFP-2A-Env encoding lentiviral vectors were used to infect lung slices in culture. Expression levels of miRNAs did not, in any of the cases, resemble lung tissue findings. Fewer than 5% of cells in lung slices were found to be infected, suggesting that changes in miRNA expression could be masked by the background of normal cells. Nevertheless, increasing JSRV21 concentration did not yield higher infection levels, indicating that those might be more dependent on the availability of JSRV’s target cells, dividing type II pneumocytes, than viral concentration. Taken together, this study has revealed new information on miRNA expression in OPA-affected sheep, including expression patterns in lung and serum. Future work should focus on developing a permissive replication system to allow the study of miRNAs in early JSRV-induced transformation events

A database and digital signal processing framework for the perceptual analysis of voice quality

Bermúdez de Alvear RM, Corral J, Tardón LJ, Barbancho AM, Fernández Contreras E, Rando Márquez S, Martínez-Arquero AG, Barbancho I. A database and digital signal processing framework for the perceptual analysis of voice quality. Pan European Voice Conferenc: PEVOC 11 Abstract Book. Aug. 31-Sept.2, 2015.Introduction. Clinical assessment of dysphonia relies on perceptual as much as instrumental methods of analysis [1]. The perceptual auditory analysis is potentially subject to several internal and external sources of bias [2]. Furthermore acoustic analyses which have been used to objectively characterize pathological voices are likely to be affected by confusion variables such as the signal processing or the hardware and software specifications [3]. For these reasons the poor correlation between perceptual ratings and acoustic measures remains to be a controversial matter [4]. The availability of annotated databases of voice samples is therefore of main importance for clinical and research purposes. Databases to perform digital processing of the vocal signal are usually built from English speaking subjects’ sustained vowels [5]. However phonemes vary from one language to another and to the best of our knowledge there are no annotated databases with Spanish sustained vowels from healthy or dysphonic voices. This work shows our first steps to fill in this gap. For the aim of aiding clinicians and researchers in the perceptual assessment of voice quality a two-fold objective was attained. On the one hand a database of healthy and disordered Spanish voices was developed; on the other an automatic analysis scheme was accomplished on the basis of signal processing algorithms and supervised learning machine techniques. Material and methods. A preliminary annotated database was created with 119 recordings of the sustained Spanish /a/; they were perceptually labeled by three experienced experts in vocal quality analysis. It is freely available under Links in the ATIC website (www.atic.uma.es). Voice signals were recorded using a headset condenser cardioid microphone (AKG C-544 L) positioned at 5 cm from the speaker’s mouth commissure. Speakers were instructed to sustain the Spanish vowel /a/ for 4 seconds. The microphone was connected to a digital recorder Edirol R-09HR. Voice signals were digitized at 16 bits with 44100 Hz sampling rate. Afterwards the initial and last 0.5 second segments were cut and the 3 sec. mid portion was selected for acoustic analysis. Sennheiser HD219 headphones were used by judges to perceptually evaluate voice samples. To label these recordings raters used the Grade-Roughness-Breathiness (GRB) perceptual scale which is a modified version of the original Hirano’s GRBAS scale, posteriorly modified by Dejonckere et al., [6]. In order to improve intra- and inter-raters’ agreement two types of modifications were introduced in the rating procedure, i.e. the 0-3 points scale resolution was increased by adding subintervals to the standard 0-3 intervals, and judges were provided with a written protocol with explicit definitions about the subintervals boundaries. By this way judges could compensate for the potential instability that might occur in their internal representations due to the perceptual context influence [7]. Raters’ perceptual evaluations were simultaneously performed by means of connecting the Sennheiser HD219 headphones to a multi-channel headphone preamp Behringer HA4700 Powerplay Pro-XL. The Yin algorithm [8] was selected as initial front-end to identify voiced frames and extract their fundamental frequency. For the digital processing of voice signals some conventional acoustic parameters [6] were selected. To complete the analysis the Mel-Frequency Cepstral Coefficients (MFCC) were further calculated because they are based on the auditory model and they are thus closer to the auditory system response than conventional features. Results. In the perceptual evaluation excellent intra-raters agreement and very good inter-raters agreement were achieved. During the supervised machine learning stage some conventional features were found to attain unexpected low performance in the classification scheme selected. Mel Frequency Cepstral Coefficients were promising for assorting samples with normal or quasi-normal voice quality. Discussion and conclusions. Despite it is still small and unbalanced the present annotated data base of voice samples can provide a basis for the development of other databases and automatic classification tools. Other authors [9, 10, 11] also found that modeling the auditory non-linear response during signal processing can help develop objective measures that better correspond with perceptual data. However highly disordered voices classification remains to be a challenge for this set of features since they cannot be correctly assorted by either conventional variables or the auditory model based measures. Current results warrant further research in order to find out the usability of other types of voice samples and features for the automatic classification schemes. Different digital processing steps could be used to improve the classifiers performance. Additionally other types of classifiers could be taken into account in future studies. Acknowledgment. This work was funded by the Spanish Ministerio de Economía y Competitividad, Project No. TIN2013-47276-C6-2-R has been done in the Campus de Excelencia Internacional Andalucía Tech, Universidad de Málaga. References [1] Carding PN, Wilson JA, MacKenzie K, Deary IJ. Measuring voice outcomes: state of the science review. The Journal of Laryngology and Otology 2009;123,8:823-829. [2] Oates J. Auditory-perceptual evaluation of disordered voice quality: pros, cons and future directions. Folia Phoniatrica et Logopaedica 2009;61,1:49-56. [3] Maryn et al. Meta-analysis on acoustic voice quality measures. J Acoust Soc Am 2009; 126, 5: 2619-2634. [4] Vaz Freitas et al. Correlation Between Acoustic and Audio-Perceptual Measures. J Voice 2015;29,3:390.e1 [5] “Multi-Dimensional Voice Program (MDVP) Model 5105. Software Instruction Manual”, Kay PENTAX, A Division of PENTAX Medical Company, 2 Bridgewater Lane, Lincoln Park, NJ 07035-1488 USA, November 2007. [6] Dejonckere PH, Bradley P, Clemente P, Cornut G, Crevier-Buchman L, Friedrich G, Van De Heyning P, Remacle M, Woisard V. A basic protocol for functional assessment of voice pathology, especially for investigating the efficacy of (phonosurgical) treatments and evaluating new assessment techniques. Guideline elaborated by the Comm. on Phoniatrics of the European Laryngological Society (ELS). Eur Arch Otorhinolaryngol 2001;258:77–82. [7] Kreiman et al. Voice Quality Perception. J Speech Hear Res 1993;36:21-4 [8] De Cheveigné A, Kawahara H. YIN, a fundamental frequency estimator for speech and music. J. Acoust. Soc. Amer. 202; 111,4:1917. [9] Shrivastav et al. Measuring breathiness. J Acoust Soc Am 2003;114,4:2217-2224. [10] Saenz-Lechon et al. Automatic Assessment of voice quality according to the GRBAS scale. Eng Med Biol Soc Ann 2006;1:2478-2481. [11] Fredouille et al. Back-and-forth methodology for objective voice quality assessment: from/to expert knowledge to/from automatic classification of dysphonia. EURASIP J Appl Si Pr 2009.Campus de Excelencia Internacional Andalucía Tech, Universidad de Málaga. Ministerio de Economía y Competitividad, Projecto No. TIN2013-47276-C6-2-R

Estimation of carbon stocks under different soil uses in the central highlands of Mexico

La pérdida de los almacenes de carbono orgánico en el suelo y su expulsión a la atmósfera por efecto de las actividades antrópicas debe entenderse como un problema que trasciende del contexto ambiental al social y económico, siendo la degradación de los suelos una de sus múltiples manifestaciones. Si bien, la acumulación del carbono en el suelo depende de manera natural de la interacción de una serie de factores bióticos y abióticos, las prácticas de manejo, con frecuencia implican el deterioro de sus propiedades físicas, químicas y biológicas, y con ello el incremento de los niveles de mineralización y reducción de este almacén. Este trabajo se realizó en el municipio de San José del Rincón, Estado de México en suelos volcánicos bajo diferentes condiciones de uso y tiempos de manejo, de los cuales se obtuvieron muestras en 11 sitios de las que se realizaron análisis físicos y químicos y se obtuvo el almacén de carbono orgánico del suelo a una profundidad de 20 cm. Los resultados obtenidos indicaron que para los suelos bajo uso agrícola y pastoreo hubo incrementos significativos en la densidad aparente, mayor acidez y disminución de la concentración de carbono, con respecto a los suelos forestales. Los almacenes de carbono orgánico para los usos de suelo forestal, agrícola y pecuario a 20 cm de profundidad fueron de 131, 53 y 63 Mg C ha-1 respectivamente, estos resultados sugieren que el tiempo e intensidad de manejo son determinantes en la pérdida de los almacenes del carbono orgánico del suelo, así como en la alteración de algunas de sus propiedades físicas y químicas.The loss of organic carbon stocks from the soil and their expulsion into the atmosphere due to the effect of anthropogenic activities must be understood as a problem that goes beyond the environmental to the social and economic context, with the soil degradation being just one of its many impacts. While the accumulation of carbon in the soil depends naturally on the interaction among a series of biotic and abiotic factors, management practices often cause the deterioration of its physical, chemical and biological properties, and, thus, increase the levels of mineralization and a reduction in carbon stocks. This study was undertaken in the municipality of San José del Rincón, State of Mexico, in volcanic soils under different conditions of land use and handling time periods. Samples were obtained at 11 plots, on which physical and chemical analyses were undertaken, obtaining soil organic carbon stocks at 20 cm top soil. The results indicated that, for the soils used for agriculture and induced pasture, there were significant increases in the bulk density, greater acidity and a decrease in carbon concentration compared to forest soils. The organic carbon stocks taken from soils used in forestry, agriculture and induced pasture at a depth of 20 cm were 131, 53 and 63 Mg C ha-1 respectively, results which suggest that the timing and intensity of management are determinants in the loss of soil organic carbon stocks from the soil, as well as the alteration of some of their physical and chemical properties.Proyecto PROMEP DSA/103.5/15/718

Desafío tecnológico: herramienta para trabajar y evaluar las competencias básicas y generales en los estudios de grado de la E.T.S.I. de Telecomunicación

This work presents the evolution of the five editions of the educational activity named “Technological Challenge” specially focused on the students at “Escuela Técnica Superior de Ingeniería de Telecomunicación” (ETSIT), but open to all students of the “Universidad de Málaga” (UMA). This initiative has been developed in the context of the educational innovation project PIE17-021 funded by UMA. The “Technological Challenge” consists on the formulation of specific real problems, which students must face in a competitive regime. This activity allows the reinforcement and evaluation of basic and general competences reached by the graduate students in the ETSIT. After nearly five years, this paper describes the evaluation of the results, regarding interest and participation of the students in the “Technology Challenge” along with the basic and general competences reached by the students.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. PIE17-02

The RR Lyrae projected density distribution from the Galactic centre to the halo

The projected density distribution of type ab RR Lyrae (RRab) stars was characterised from the innermost regions of the Milky Way to the halo, with the aim of placing constraints on the Galaxys evolution. The compiled sample (N_RRab = 64,850) stems from fundamental mode RR Lyrae variables identified by the VVV, OGLE, and Gaia surveys. The distribution is well fitted by three power laws over three radial intervals. In the innermost region (R < 2.2 deg) the distribution follows Sigma_RRab[1] propto R ^(-0.94 +- 0.051), while in the external region the distribution adheres to Sigma_RRab[2] propto R^(-1.50 +- 0.019) for 2.2 deg< R <8.0 deg and Sigma_RRab[3] propto R ^(-2.43 +- 0.043) for 8.0 deg < R <30.0 deg. Conversely, the cumulative distribution of red clump (RC) giants exhibits a more concentrated distribution in the mean, but in the central R < 2.2 deg the RRab population is more peaked, whereas globular clusters (GCs) follow a density power law (Sigma_GCs propto R ^(-1.59 +- 0.060) for R<30.0 deg) similar to that of RRab stars, especially when considering a more metal-poor subsample ([Fe/H]<-1.1 dex). The main conclusion emerging from the analysis is that the RRab distribution favours the star cluster infall and merger scenario for creating an important fraction (>18 %) of the central Galactic region. The radii containing half of the populations (half populations radii) are R_H=6.8 deg (0.99 kpc), R_H =4.2 deg (0.61 kpc), and R_H =11.9 deg (1.75 kpc) for the RRab stars, RC giants, and GCs, respectively. Finally, merely 1% of the stars have been actually discovered in the innermost region (R < 35 pc) out of the expected (based on our considerations) total number of RRab therein: N sim 1,562. That deficit will be substantially ameliorated with future space missions like the Nancy Grace Roman Space Telescope (formerly WFIRST).Comment: 11 pages, 9 figures and 2 tables. Accepted for publication in A&

PLANEACIÓN DIDÁCTICA GENERAL DE LA ASGNATURA: DESARROLLO PERSONAL

GUÍA DIDÁCTICA / PLANEACIÓN DIDÁCTICA (NMS

Laboratorio remoto eLab3D: un entorno innovador para el aprendizaje de competencias prácticas en electrónica

Este artículo muestra la utilización de las nuevas tecnologías al servicio del aprendizaje de competencias prácticas en electrónica, siendo un ejemplo de adaptación de los recursos educativos a diferentes contextos y necesidades. Se trata de un laboratorio remoto en el que la adecuada complementación de un hardware configurable y un software de última generación permite al estudiante realizar prácticas de electrónica y diseño de circuitos en un mundo virtual 3D. El usuario dispone de un avatar e interacciona con réplicas virtuales de instrumentos, placas de circuitos, componentes o cables de forma muy similar a como se opera en un laboratorio presencial. Pero lo realmente destacable es que el usuario manipula instrumentación y circuitos que están ubicados en un laboratorio real. Todo ello se ha conseguido con un sistema escalable y de bajo coste. Finalizado el diseño y desarrollo de la plataforma se han realizado las primeras pruebas con estudiantes, profesores y profesionales para valorar su percepción respecto al uso de eLab3D, obteniéndose unos resultados muy positivos

Polyphenols and IUGR Pregnancies : effects of the Antioxidant Hydroxytyrosol on Brain Neurochemistry and Development in a Porcine Model

Supplementation of a mother's diet with antioxidants, such as hydroxytyrosol (HTX), has been proposed to ameliorate the adverse phenotypes of fetuses at risk of intrauterine growth restriction. In the present study, sows were treated daily with or without 1.5 mg of HTX per kilogram of feed from day 35 of pregnancy (at 30% of total gestational period), and individuals were sampled at three different ages: 100-day-old fetuses and 1-month- and 6-month-old piglets. After euthanasia, the brain was removed and the hippocampus, amygdala, and prefrontal cortex were dissected. The profile of the catecholaminergic and serotoninergic neurotransmitters (NTs) was characterized and an immunohistochemical study of the hippocampus was performed. The results indicated that maternal supplementation with HTX during pregnancy affected the NT profile in a brain-area-dependant mode and it modified the process of neuron differentiation in the hippocampal CA1 and GD areas, indicating that cell differentiation occurred more rapidly in the HTX group. These effects were specific to the fetal period, concomitantly with HTX maternal supplementation, since no major differences remained between the control and treated groups in 1-month- and 6-month-old pigs

Implantación de prácticas de Ecodiseño en las empresas. aplicación en el rediseño diseño de un accesorio para sillas de ruedas

En la actualidad se están incrementando los diseños basados en el desarrollo sostenible en el campo del Diseño Industrial. La meta es aumentar el desempeño ambiental de los productos de las empresas. Para ello hay que identificar y desarrollar estrategias en la ingeniería de producto para lograr un diseño respetuoso con el medioambiente. El ecodiseño incorpora estos criterios en la concepción y desarrollo de un producto. Esta metodología toma en consideración el comportamiento ambiental del producto a lo largo de su ciclo de vida, además de suponer una ventaja tecnológica y económica para la empresa. Pero a pesar de los beneficios en su uso y la concienciación de las empresas en disminuir los impactos ambientales, el ecodiseño está encontrando dificultades en su implantación. El objetivo de este trabajo es entender las barreras que se encuentran las empresas para poder aplicar el ecodiseño. Además de conocer los distintos niveles de aplicación y de las herramientas se pueden apoyar en función de la sistematización del ecodiseño. Tras la investigación se desarrollará unas pautas para eliminar los obstáculos que se encuentran las empresas y facilitar la implantación de las prácticas de ecodiseño. En este artículo se muestra la aplicación de la práctica de ecodiseño en el rediseño de un sistema de desplazamiento para sillas de ruedas autopropulsables dirigida a personas con movilidad reducida. En esta práctica, se utilizan las herramientas adecuadas para conseguir disminuir los impactos ambientales durante todo su ciclo de vistas sin olvidar el resto de las características que debe tener el producto para su comercialización. En el diseño se han tenido presente los aspectos técnicos, procesos productivos y aspectos económicos, para que se garanticen su eficiencia. Se han realizado varios prototipos para comprobar y corregir los fallos del diseño y demostrar su funcionalidad