Expression of Brugmansia candida Hyoscyamine 6beta-Hydroxylase gene in Saccharomyces cerevisiae and its potential use as biocatalyst

<p>Abstract</p> <p>Background</p> <p>Tropane alkaloids, mainly hyoscyamine and scopolamine, are widely used in medicine due to their anticholinergic activity. Scopolamine has a higher demand being the more valuable alkaloid due to its fewer side effects and higher physiological activity. Anisodamine (6β-hydroxyhyoscyamine) is the intermediate in the conversion of hyoscyamine into scopolamine. Current studies report that this alkaloid is potentially applicable in medicine. The gene that codifies for Hyoscyamine 6-β hydroxylase, the enzyme responsible for hyoscyamine hydroxylation and epoxidation, leading to scopolamine was isolated from <it>Brugmansia candida</it>.</p> <p>Results</p> <p>The <it>h6h</it>cDNA was cloned into pYES2.1 and pYES2.1/V5-His-TOPO vectors to produce an untagged and a tagged protein, respectively. The H6H enzyme was produced in <it>Saccharomyces cerevisiae </it>in order to obtain a biological catalyst for potential industrial applications. Protein extracts of the induced yeast were analyzed by Western blot. The expression was detected 4 h after induction and no degradation was observed during the period assayed. The tagged and the untagged proteins were able to transform hyoscyamine, showing a functional expression of the <it>h6h</it>cDNA.</p> <p>Conclusion</p> <p>The strains obtained in this work are promising and potentially applicable in biocatalytic processes.</p

Ripple Effects of Surface acting: A Diary Study among Dual-earner Couples

This study among 80 dual-earner couples examines the ripple effects of emotional labour – on a daily basis. Specifically, we propose that employees who engage in surface acting at work drain their energetic resources, and undermine their own relationship satisfaction. Drawing upon conservation of resources (COR) theory, we predicted that work-related exhaustion would mediate the relationship between surface acting at work and at home. In addition, we hypothesized that employees’ emotional energy in the evening would mediate the relationship between surface acting at home and (actor and partner) satisfaction with the relationship. Participants filled in a survey and a diary booklet during five consecutive working days (N = 80 couples, N = 160 participants x 5 days, N = 800 occasions). The hypotheses were tested with multilevel analyses, using the actor–partner interdependence model. Results showed that daily work-related exhaustion partially mediated the relationship between daily surface acting at work and at home. As hypothesized, daily surface acting at home influenced own and partner’s daily relationship satisfaction through reduced daily emotional energy. These findings offer support for COR theory, and have important implications for organizations that encourage emotion regulation

Formation of secondary organic aerosols from the ozonolysis of dihydrofurans

In this work we report the study of the ozonolysis of 2,5-dihydrofuran and 2,3-dihydrofuran and the reaction conditions leading to the formation of secondary organic aerosols. The reactions have been carried out in a Teflon chamber filled with synthetic air mixtures at atmospheric pressure and room temperature. The ozonolysis only produced particles in the presence of SO2. Rising relative humidity from 0 to 40% had no effect on the production of secondary organic aerosol in the case of 2,5-dihydrofuran, while it reduced the particle number and particle mass concentrations from the 2,3-dihydrofuran ozonolysis. The water-to-SO2 rate constant ratio for the 2,3-dihydrofuran Criegee intermediate was derived from the secondary organic aerosol (SOA) yields in experiments with different relative humidity values, kH2O/kSO2 = (9.8 ± 3.7) × 10-5. The experimental results show that SO3 may not be the only intermediate involved in the formation or growth of new particles in contrast to the data reported for other Criegee intermediate-SO2 reactions. For the studied reactions, SO2 concentrations remained constant during the experiments, behaving as a catalyst in the production of condensable products. Computational calculations also show that the stabilised Criegee intermediates from the ozonolysis reaction of both 2,5-dihydrofuran and 2,3-dihydrofuran may react with SO2, resulting in the regeneration of SO2 and the formation of low-volatility organic acids.Instituto de Investigaciones Fisicoquímicas Teóricas y Aplicada

Retinal pigment epithelium degeneration caused by aggregation of PRPF31 and the role of HSP70 family of proteins

Background Mutations in pre-mRNA splicing factor PRPF31 can lead to retinitis pigmentosa (RP). Although the exact disease mechanism remains unknown, it has been hypothesized that haploinsufficiency might be involved in the pathophysiology of the disease. Methods In this study, we have analyzed a mouse model containing the p.A216P mutation in Prpf31 gene. Results We found that mutant Prpf31 protein produces cytoplasmic aggregates in the retinal pigment epithelium and decreasing the protein levels of this splicing factor in the nucleus. Additionally, normal protein was recruited in insoluble aggregates when the mutant protein was overexpressed in vitro. In response to protein aggregation, Hspa4l is overexpressed. This member of the HSP70 family of chaperones might contribute to the correct folding and solubilization of the mutant protein, allowing its translocation to the nucleus. Conclusions Our data suggests that a mechanism haploinsufficiency and dominant-negative is involved in retinal degeneration due to mutations in PRPF31. HSP70 over-expression might be a new therapeutic target for the treatment of retinal degeneration due to PRPF31 mutations.This project has been financed through a) The ISCIII (Miguel Servet-I, 2015), co-financed by the European Regional Development Fund (ERDF), No CP15/00071. b) The European Union’s Horizon 2020 research and innovation program, under grant agreement No 634479. c) Regional Ministry of Economy, Innovation and Science of the Junta de Andalucía, No P09-CTS-04967.info:eu-repo/semantics/publishedVersio

Functional characterization of correct and incorrect feature integration

Advance access publication date 30 April 2022Our sensory system constantly receives information from the environment and our own body. Despite our impression to the contrary, we remain largely unaware of this information and often cannot report it correctly. Although perceptual processing does not require conscious effort on the part of the observer, it is often complex, giving rise to errors such as incorrect integration of features (illusory conjunctions). In the present study, we use functional magnetic resonance imaging to study the neural bases of feature integration in a dual task that produced ~30% illusions. A distributed set of regions demonstrated increased activity for correct compared to incorrect (illusory) feature integration, with increased functional coupling between occipital and parietal regions. In contrast, incorrect feature integration (illusions) was associated with increased occipital (V1–V2) responses at early stages, reduced functional connectivity between right occipital regions and the frontal eye field at later stages, and an overall decrease in coactivation between occipital and parietal regions. These results underscore the role of parietal regions in feature integration and highlight the relevance of functional occipito-frontal interactions in perceptual processing.This work was supported by the Spanish Ministry of Science and Innovation research projects PSI2017-88136 and PID2020-119033GB-I00 and the local government of Andalusia (Proyectos de I+D+i en el marco del Programa Operativo FEDER, B-SEJ-570-UGR20) to Ana B. Chica. Pedro M. Paz- Alonso was supported by grants from the Spanish Ministry of Science and Innovation [RYC-2014- 15440 and PGC2018-093408-B-I00], Neuroscience projects from the Fundación Tatiana Pérez de Guzmán el Bueno, Basque Government [PIBA-2021-1-0003], and a grant from “la Caixa” Banking Foundation under the project code LCF/PR/HR19/52160002. BCBL acknowledges support by the Basque Government through the BERC 2022-2025 program and by the Spanish State Research Agency through BCBL Severo Ochoa excellence accreditation CEX2020-001010-S

Innovative Tourism Clusters: Myth or Reality? Empirical Evidence from Benidorm

Paper submitted to TCVT3 Bozen/Bolzano, 10-12 April 2014, International workshop on Tourists as Consumers, Visitors, TravellersThis paper analyses the applicability of the cluster concept to tourist destinations, taking Benidorm as the reference for study: a destination on the Mediterranean coast of Spain which has attained a high degree of competitive success with the sun and sand product in which it has specialized. The main result obtained is that Benidorm's success is not derived from or favoured by the existence of a cluster in the destination, since important elements are present that prevent affirmation of its existence. This paper is the preliminary result of a research project that has been carried out within the framework of the project "Methodology, criteria and impementation of the cluster theory in consolidated tourism areas: innovation, competitiveness and territorial synergies" under the Spanish National R&D&I Plan 2008-2011 supported by the Ministry of Science and Innovation.This research has been carried out within the framework of the project “Methodology, criteria and implementation of the cluster theory in consolidated tourism areas: innovation, competitiveness and territorial synergies” under the Spanish National R&D&I Plan 2008-2011 supported by the Ministry of Science and Innovation

Economic crises and tourism competitiveness: A Markov swtiching regression approach

Authors discuss the effects that economic crises generate on the global market shares of tourism destinations, through a series of potential transmission mechanisms based on the main economic competitiveness determinants identified in the previous literature using a non-linear approach. Specifically a Markov Switching Regression approach is used to estimate the effect of two basic transmission mechanisms: reductions of internal and external tourism demands and falling investment

Ensayos experimentales para la obtención del coeficiente hidráulico como un factor relevante en el desarrollo del proyecto de un buque

12th International Conference on Project Engineering[Resumen]En este artículo se presenta un estudio experimental del comportamiento de un modelo sujeto a la inundación progresiva de sus compartimentos. El estudio consta de varios ensayos mediante los que se inunda uno o varios compartimientos utilizando aberturas circulares, y de otras formas, con diferentes dimensiones. En cada ensayo se mide en el dominio del tiempo el calado y los ángulos de escora y cabeceo. Teniendo en cuenta estos movimientos se estima la cantidad de agua y caudal en el interior del compartimiento. Estos datos sirven para obtener el coeficiente hidráulico asociado a las aberturas del modelo. Estos coeficientes pueden ser utilizados posteriormente para calibrar programas de cálculo de la inundación progresiva en los buques. ------ [abstract] In this paper an experimental study appears about the behavior of a model subject to the progressive flood of its compartments. The study consists of several tests by means of which one or more compartments are flooded using circular openings, and of other forms, with different dimensions. In every test the domain of the time the draw and the angles of roll and pitch are measured. Bearing these movements in mind the volume and water quantity is estimated inside the compartment. This information serves to obtain the hydraulic coefficient associated with the openings of the model. These coefficients can be used later to calibrate programs of calculation to the progressive flooding in the ships