Promoter trapping in microalgae using the antibiotic paromomycin as selective agent

The lack of highly active endogenous promoters to drive the expression of transgenes is one of the main drawbacks to achieving efficient transformation of many microalgal species. Using the model chlorophyte Chlamydomonas reinhardtii and the paromomycin resistance APHVIII gene from Streptomyces rimosus as a marker, we have demonstrated that random insertion of the promoterless marker gene and subsequent isolation of the most robust transformants allows for the identification of novel strong promoter sequences in microalgae. Digestion of the genomic DNA with an enzyme that has a unique restriction site inside the marker gene and a high number of target sites in the genome of the microalga, followed by inverse PCR, allows for easy determination of the genomic region, which precedes the APHVIII marker gene. In most of the transformants analyzed, the marker gene is inserted in intragenic regions and its expression relies on its adequate insertion in frame with native genes. As an example, one of the new promoters identified was used to direct the expression of the APHVIII marker gene in C. reinhardtii, showing high transformation efficiencies.Junta de Andalucía (P09-CVI-5053)Junta de Andalucía (BIO-214

Propuesta de un nuevo modelo de prácticum en el título de Grado de Maestro de Primaria de la Universidad de Alicante

Comunicación presentada en el XI Symposium Internacional sobre el practicum y las prácticas en empresas en la formación universitaria "Evaluación y supervisión del practicum: el compromiso con la calidad de las practicas", Poio (Pontevedra), 27-29 junio 2011.El presente artículo pretende ofrecer nuestro punto de vista sobre el desarrollo de la asignatura Prácticum del título de grado de Maestro de Primaria, tomando como modelo lo dispuesto en el plan de estudios de la Facultad de Educación Alicante. En él tendrán cabida, tanto los contenidos y su secuenciación, como los roles asignados a los agentes implicados en el desarrollo de la asignatura. Ofreceremos nuestra visión particular del tratamiento de la evaluación y terminaremos justificando como la integración de las TIC pueden ser utilizadas como medio de información, comunicación y aprendizaje a través de un Entorno Virtual de Aprendizaje (EVA) y un diseño específico a través de herramientas basadas en la Web 2.0, Personal Learning Environments (PLE)

Numerical and experimental evaluation of dielectric properties of thermally aged insulating paper used in power transformers

Due to the relevant role of the cellulosic insulation in the dielectric system of a power transformer, it is necessary to know its dielectric properties and its evolution as insulating material. The dependence of the dielectric properties of solid insulation on the moisture content, temperature, or pressure is well-defined. However, it is not clear the effect of the paper degradation on its insulating capacity. To study the impact of ageing on the dielectric properties, samples of Kraft paper were subjected to an accelerated hygrothermal ageing. Degree of polymerization (DP) value was reduced from 1000 to 200, with intermediate study points. All samples were conditioned to the same moisture content in order to only evaluate the effect of the paper degradation. Different dielectric properties, such as loss factor (tanδ) and complex permittivity (ε) were measured using the dielectric spectroscopy technique. These experimental results were used to develop a numerical model with the finite element-based tool Comsol Multiphysics. This model provided some extra information about the material properties, such as the electric field distribution. Results showed that the degradation of Kraft paper modifies its dielectric response.We acknowledge the support of the Spanish Ministry of Science and Innovation by means of the National Research Project Asset management of biodegradable-fluid-based transformers (PID2019-107126RB-C22/AEI/ 10.13039/501100011033). The authors also wish to thank “Fundación Iberdrola” for its financial support for the research project: S090 “Análisis de las Propiedades Dieléctricas de Aislamientos Sólidos Impregnados con Líquidos Dieléctricos”. C. Méndez also wants to acknowledge the Spanish Ministry of Science, Innovation and Universities for the financial support for the FPU grant (FPU19/01849)

Propuesta de mantenimiento productivo total para mejorar la eficiencia de la línea de envasado hotfill en una empresa de bebidas no alcohólicas

La investigación presentada se orientó a la propuesta de mejora de la eficiencia de una línea de envasado hotfill en una empresa productora y comercializadora de bebidas no alcohólicas, a través de la aplicación de las herramientas de la metodología del mantenimiento productivo total (TPM). Se empezó realizando un análisis acerca de la demanda externa, regional y local de las bebidas hotfill, donde se evidenciaba un constante crecimiento, obligando al área comercial en que las metas vayan acordes al mercado. Sin embargo, el área operativa no lograba cumplir con ello, ocasionando se realicen horas extras de trabajo y aparecieran costos no proyectados. Se desarrolló el análisis de los datos históricos de la empresa, identificándose tres grandes problemas: indisponibilidad de máquinas, bajo rendimiento de los equipos y bajo nivel del cumplimiento de las órdenes de trabajo de mantenimiento. Para cumplir los objetivos principales y específicos planteados, se propone el uso de herramientas con un enfoque en la metodología del TPM; centrándose en 3 pilares: Mantenimiento planificado, autónomo, y actividades de departamentos administrativos y de apoyo. En conclusión, se logró obtener los siguientes indicadores: Incremento en la disponibilidad de equipos del 12%; en el rendimiento de equipos del 4% y en el nivel de cumplimiento de órdenes de trabajo de mantenimiento del 15%. Por consiguiente, a través de la propuesta del TPM se logró mejorar la eficiencia de la línea de envasado hotfill en una empresa de bebidas no alcohólicas a un 25% vs el escenario anterior

Multidisciplinary Ophthalmic Imaging Visible Versus Near-Infrared Optical Performance of Diffractive Multifocal Intraocular Lenses

PURPOSE. The purpose of this study was to compare the optical performance of diffractive multifocal intraocular lenses (DMIOLs) with visible (VIS) illumination with that of near infrared (NIR) illumination, the latter being used to test pseudophakic eyes in clinical aberrometers and double-pass systems. METHODS. Two DMIOLs of different design (Tecnis þ2.75 D ZKB00 and AcrySof þ2.5 D SV25T0) were tested in vitro in a model eye under both VIS (k ¼ 530 nm) and NIR (k ¼ 780 nm) illumination, and variations in the add power of the lenses were determined. Moreover, for the two wavelengths, the energy efficiency and modulation transfer function at the DMIOLs' far and near foci were measured with pupils of 3.0 and 4.5 mm. Two counterpart monofocal IOLs (Tecnis ZA9003 and AcrySof SN60WF) were included as references in the comparison. RESULTS. With VIS light, the two DMIOLs produced relatively well-contrasted images at their near and far foci. Under NIR illumination, the add power increased, whereas the energy efficiency of the near focus decreased and that of far focus increased. Hence, the DMIOLs tended to behave like monofocal lenses because they generated good quality well-contrasted images only at their far foci. CONCLUSIONS. In addition to changes in add power, the optical performances of the DMIOLs measured under either VIS or NIR illumination are considerably different. Whereas they show two distinct (near and far) foci under VIS light, their optical performances under NIR illumination are clearly biased in favor of their far focus. These results may help prevent a misleading use of NIR-based clinical instruments for the assessment of eyes implanted with DMIOLs. Keywords: aberrometers, cataract surgery, diffractive multifocal intraocular lens, intraocular lens, pseudophakic eye, visual function testing R emoval of the crystalline lens followed by implantation of a multifocal intraocular lens (MIOL) is presently a surgical procedure primarily aimed at providing pseudoaccommodation to patients after cataract surgery. Diffractive MIOLs (DMIOLs) have proven to provide reliable and better clinical outcomes than their refractive MIOL counterparts or accommodating IOLs, 1 so there is a growing interest in assessing their optical performance in vivo. To objectively determine the imaging quality of DMIOLs in patients, several authors have recently used both wavefront aberrometers 2,3 and double-pass-based systems. Because the wavelengths of NIR testing are quite different from the wavelength corresponding to the maximum value of the photopic sensitivity of the eye under VIS spectrum (approximately 550 nm), for which DMIOLs are designed, there is a mismatch between the NIR and VIS wavelengths, which gives rise to the following issues: -A change in add power of the DMIOL between VIS and NIR (the larger the NIR wavelength, the larger the measured add power), 12 which in turn, has an effect on halo formation and size. 14,15 -A change in distribution of energy between the foci of the DMIOL. METHODS DMIOL Characteristics The Tecnis ZKB00 DMIOL, with þ2.75 diopter (D) add power, has an aspheric anterior surface. The diffractive profile covers the full aperture of the lens and consists of 15 diffractive rings with step boundaries of the same height, intended for approximately equal light distribution between the far and near foci, independent of pupil size. The wavefront-designed aspheric optics of this DMIOL produce a maximum spherical aberration (SA) of À0.27 lm for a 6.0-mm pupil. On the other hand, the AcrySof ReSTOR SV25T0 DMIOL with þ2.5 D add power has an anterior apodized diffractive surface (3.4-mm diameter) within which there is a central refractive zone (1.0 mm diameter approximately). The outer region of the lens to the 6-mm edge is purely refractive, and thus, the central and outer refractive parts of the lens are intended for distance vision. In addition, the diffractive area presents seven concentric rings with step boundaries of decreasing height, which allows for an asymmetrical and pupil-dependent light distribution between the far and near foci that benefits the far focus for large pupils. Additionally, the reference monofocal Tecnis ZA9003 (AMO Groningen) and AcrySof SN60WF (Alcon Laboratories) IOLs were included in our study. We emphasize that each monofocal IOL shares the same aspherical design and the same material, with its diffractive counterpart. Thus, the lenses of each pair (DMIOL and monofocal counterpart) have similar characteristics regarding the compensation for high order aberration (mainly SA) and the spectral variation of the refractive index. All studied lenses had a base optical power of 20 D, which in the case of the DMIOLs corresponded to distance focus. Experimental Setup for Optical Imaging Quality Assessment Assessment of the optical imaging quality of the IOLs was made using an optical test bench with a model eye (artificial cornea plus wet cell) that has been described in detail elsewhere. 26,27 Instead, we used a double convex lens that provided a level of SA at the IOL plane of þ0.27 lm for a 6.0-mm pupil, 28 similar to the one induced by the human cornea on average. Energy Assessment and MTF Measurements by Image Analysis The method of characterizing the energy distribution at the focal planes of a DMIOL has been reported in detail elsewhere Briefly, the image in a given focal plane consisted of the core sharp image of the pinhole object surrounded by a blurred halo-shaped background. This background is principally formed from an out-of-focus image produced by the other focus of the MIOL but may have additional contributions from a variety of factors such as the energy expended in higher diffraction orders, 13 scattering produced by the diffractive steps of the lens

Dopaminergic and glutamatergic signaling crosstalk in Huntington's disease neurodegeneration : the role of p25/cyclin-dependent kinase 5

Altered glutamatergic and dopaminergic signaling has been proposed as contributing to the specific striatal cell death observed in Huntington's disease (HD). However, the precise mechanisms by which mutant huntingtin sensitize striatal cells to dopamine and glutamate inputs remain unclear. Here, we demonstrate in knock-in HD striatal cells that mutant huntingtin enhances dopaminemediated striatal cell death via dopamineD1 receptors. Moreover, we show thatNMDAreceptors specifically potentiate the vulnerability of mutant huntingtin striatal cells to dopamine toxicity as pretreatment withNMDAincreased D1R-induced cell death in mutant but not wild-type cells. As potential underlying mechanism of increased striatal vulnerability, we identified aberrant cyclin-dependent kinase 5 (Cdk5) activation. We demonstrate that enhanced Cdk5 phosphorylation and increased calpain-mediated conversion of the Cdk5 activator p35 into p25 may account for the deregulation of Cdk5 associated to dopamine and glutamate receptor activation in knock-in HD striatal cells. Moreover, supporting a detrimental role of Cdk5 in striatal cell death, neuronal loss can be widely prevented by roscovitine, a potent Cdk5 inhibitor. Significantly, reduced Cdk5 expression together with enhanced Cdk5 phosphorylation and p25 accumulation also occurs in the striatum of mutant HdhQ111 mice and HD human brain suggesting the relevance of deregulated Cdk5 pathway in HD pathology. These findings provide new insights into the molecular mechanisms underlying the selective vulnerability of striatal cells in HD and identify p25/Cdk5 as an important mediator of dopamine and glutamate neurotoxicity associated to HD

Spitzer Uncovers Active Galactic Nuclei Missed by Optical Surveys in 7 Late-type Galaxies

We report the discovery using Spitzers high resolution spectrograph of 7 Active Galactic Nuclei (AGN) in a sample of 32 late-type galaxies that show no definitive signatures of AGN in their optical spectra. Our observations suggest that the AGN detection rate in late-type galaxies is possibly 4 times larger than what optical spectroscopic observations alone suggest. We demonstrate using photoionization models with an input AGN and an extreme EUV-bright starburst ionizing radiation field that the observed mid-infrared line ratios cannot be replicated unless an AGN contribution, in some cases as little as 10% of the total galaxy luminosity, is included. These models show that when the fraction of the total luminosity due to the AGN is low, optical diagnostics are insensitive to the presence of the AGN. In this regime of parameter space, the mid-infrared diagnostics offer a powerful tool for uncovering AGN missed by optical spectroscopy. The AGN bolometric luminosities in our sample range from ~3 X 10^41 - ~2 X 10^43 ergs s^-1, which, based on the Eddington limit, corresponds to a lower mass limit for the black hole that ranges from ~3 X 10^3Mdot to as high as ~1.5 X 10^5Mdot. These lower mass limits however do not put a strain on the well-known relationship between the black hole mass and the host galaxy's stellar velocity dispersion established in predominantly early-type galaxies. Our findings add to the growing evidence that black holes do form and grow in low-bulge environments and that they are significantly more common than optical studies indicate.Comment: 8 figures, 17 pages, astro-ph\0801.2766 (Abel & Satyapal 2008; ApJ accepted) and this posting designed to form a two-part investigatio

Is proteinuria a rare condition in apparently healthy and sick cats? A feline practice experience (2007–2018)

Background: Proteinuria is assumed to be less frequent in cats than in dogs and is mainly associated with chronickidney disease (CKD).Aim: The current study aimed to evaluate and compare urine protein-to-creatinine (UPC) values retrospectively in cats visited for comprehensive annual health check or for presenting systemic clinical signs related to CKD.Methods: UPC ratio was retrospectively evaluated in 112 owned cats, out of which 51 (45.5%) were apparently healthy cats according to their owners who visited for comprehensive annual health checks and 61 (54.5%) sick cats, presenting systemic clinical signs suggesting CKD, such as weight loss or polyuria/polydipsia, among others.Results: Based on UPC, the present study found that 54.5% of all cats included were borderline proteinuric or proteinuric, having increased UPC (UPC ≥ 0.2), with 35.7% included in the sick group and 18.7% in the healthcheckgroup. Increased UPC was also statistically associated with azotemia and isosthenuria (urinary-specific gravitybetween 1,008 and 1,035) in both sick and health-check groups of cats.Conclusion: Independent of the reason for their medical visit, it could be concluded that borderline proteinuria andproteinuria were statistically mainly related to CKD in cats. Furthermore, the measurement of UPC could be veryuseful in the detection and management of CKD in apparently healthy cats during a medical visit for annual health check irrespective of the age

Evaluación de la frescura del bocachico (prochilodus reticulatus) - utilizando el panel organoléptico, la trimetilamina y el recuento bacteriano

En este estudio se tomaron poblaciones de diferentes lugares: a las cuales se le hizo una evaluación de frescura, tomándose como factores indicativos para esta: La trimetilamina el panel organoléptico y el recuento bacteriano. Basados en los resultados obtenidos se determinaron los límites de aceptabilidad que la especie alcanzaba y la estimación observada nos permitió determinar cuál era apta y cual no para el consumo. Para el control de los datos de las poblaciones, se utilizaron los gráficos de dispersión, aplicando el método de los mínimos cuadrados y estableciendo el coeficiente de correlación correspondiente en cada una de las gráficas obtenidas. Se han evaluado muchos de nuestros recursos pesqueros, pero no se tiene un conocimiento de que tan duraderos son estos, aplican do como método de conservación, la refrigeración después de capturado. En el desarrollo de este trabajo se establecieron todos los parámetros posibles para observar el comportamiento de la especie Bocachico (prochilodus reticulatus Magdalena) a preservar, desde el momento de su captura hasta que llega a manos del consumidor. En este estudio te tomaron poblaciones de diferentes lugares: a las cuales se le hizo una evaluación de frescura, tomándose como factores indicativos para constar, La trimetilaminal el panel organoléptico y el recuento bacteriano. Basados en los resultados obtenidos se determinaron los límites de aceptabilidad que la especie alcanzaba y la estimación observada nos permitió determina la estructura si era apta y cual no para el consumo. Para el control de los datos de las poblaciones, se utiliza ron los gráficos de dispersión, aplicando el método de los mínimos cuadrados y estableciendo el coeficiente de correlación correspondiente en cada una de las gráficas obtenidas

Evaluation of the hypocholesterolemic effect and prebiotic activity of a lentil (Lens culinaris Medik) extract

The aim of our work was to produce a hydroalcoholic extract of lentils and to examine: a) the hypocholesterolemic action in an animal model, by studying the plasma cholesterol level and the concentration of bile acids in the faeces; b) the potential prebiotic effect, by conducting an in vitro culture fermentation experiment and assessing the level of short chain fatty acids (SCFAs) in the faeces of rats