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Inherited platelet disorders are a heterogeneous group of rare diseases, caused by inherited defects in platelet production and/or function. Their genetic diagnosis would benefit clinical care, prognosis and preventative treatments. Until recently, this diagnosis has usually been performed via Sanger sequencing of a limited number of candidate genes. High-throughput sequencing is revolutionizing the genetic diagnosis of diseases, including bleeding disorders. We have designed a novel high-throughput sequencing platform to investigate the unknown molecular pathology in a cohort of 82 patients with inherited platelet disorders. Thirty-four (41.5%) patients presented with a phenotype strongly indicative of a particular type of platelet disorder. The other patients had clinical bleeding indicative of platelet dysfunction, but with no identifiable features. The high-throughput sequencing test enabled a molecular diagnosis in 70% of these patients. This sensitivity increased to 90% among patients suspected of having a defined platelet disorder. We found 57 different candidate variants in 28 genes, of which 70% had not previously been described. Following consensus guidelines, we qualified 68.4% and 26.3% of the candidate variants as being pathogenic and likely pathogenic, respectively. In addition to establishing definitive diagnoses of well-known inherited platelet disorders, high-throughput sequencing also identified rarer disorders such as sitosterolemia, filamin and actinin deficiencies, and G protein-coupled receptor defects. This included disease-causing variants in DIAPH1 (n=2) and RASGRP2 (n=3). Our study reinforces the feasibility of introducing high-throughput sequencing technology into the mainstream laboratory for the genetic diagnostic practice in inherited platelet disorders., This study was supported by research grants from the Gerencia Regional de Salud (GRS 1370/A/16), ISCIII & Feder (PI14/01956), CIBERER CB15/00055, Fundación Séneca (19873/GERM/15) and Sociedad Española de Trombosis y Hemostasia (SETH). SPW holds a British Heart Foundation chair.

Olive mill waste has been used as sourcing materials for the isolation of humic like substances (OMW-HLS) which have demonstrated its capacity to expand the range of applicability of photo-Fenton process to pH=5. During the isolation process, membranes of three different pore sizes (300 kDa, 150 kDa and 50 kDa) were employed in order to obtain three batches of OMW-HLS. Four pollutants contained in 2013/39/EC were used as target substances: terbutryn (TBT), diclofenac (DCF), chlorfenvinphos (CVF) and pentachlorophenol (PCP). Results showed that OMW-HLS was able to enhance photo-Fenton at pH = 5, but differences were not significant, either among fractions or with commercial humic substances. Reactions were scaled-up and driven under real sunlight and pollutants removal was faster in the presence of OMW-HLS. Toxicity was monitored according to bioassays based on different organisms or cell lines. Detoxification was observed with and without OMW-HLS, although higher toxicity was detected in the presence of humic acids, most probably due to the surfactant effect, that allows a better contact between pollutant and organism.

The taxonomic status ofDendroctonus viteiWood (Coleoptera: Curculionidae: Scolytinae) andDendroctonus mexicanusHopkins is supported by biological and morphological data. However, the differentiation of these species has been extremely difficult because most of the morphological characters have not been well illustrated and some of them are geographically variable. Seminal rod shape has been the only reliable characteristic to identify these species. The main purpose of this study was to look for new morphological characters for the identification of these species as well as to evaluate geographic variation of those previously reported characters in order to produce a comprehensive redescription ofD. vitei. A total of 33 morphological characters from antenna, stridulatory apparatus, male and female genitalia, and external morphology were compared between the similar speciesD. viteiandD. mexicanus, and we identified five novel diagnostic characters and confirmed previously reported characters that differentiate these two species from each other and from other species in theDendroctonus frontalisZimmermann complex (sensu stricto). Furthermore, these characters and those of the seminal rod allowed us to re-evaluate some previously reported records for other species to broaden the known distribution ofD. vitei.

Effective responses to the impacts of climate change require the recognition that people conceptualize and experience environmental changes differently, and require the support of a range of global-to-local interdisciplinary efforts that allow a dialog between the biophysical and social sciences. In this study, we use a hybrid epistemological framework that integrates scientifc and local knowledge systems, methodological approaches, and geographic scopes to: 1) shed light on climate change in the equatorial Andes as reported by scientific and local knowledge systems, and 2) understand the role that climate factors play on land use and agricultural change in natural resource dependent communities in the region. We analyzed weather station (n = 5) data and downscaled climate data using parametric and non-parametric statistical tests, and spatial analysis techniques to detect spatio-temporal climate trends between 1965 and 2013. We also analyzed climate variability in the past four decades using qualitative information derived from a semi-structured survey (n = 36) and life history smallholder interviews (n = 8) collected in three research sites. Our study reveals significant warming trends in the region which is corroborated by local reports. Precipitation records show high inter-annual variability, with no significant decreasing or increasing trends, which contradicts people’s views in two of our research sites. Our study shows that although climate change appears to be a factor that influences farmers’ agricultural and livestock-production practices, other non-climatic forces play a more important role in determining their intensity and adoption. Despite site-level divergences and convergences, scientific and local knowledge systems can be woven together to expand our understanding of the physical and human manifestations of climate change.

This article explores the concepts involved in the term territorio , the nature of its uses and the interpretations given to it by different user groups in Latin America. In so doing, the authors find that despite its ubiquitous nature, polysemic use and its hybrid conceptual construction, territorio is a specific Latin American concept that can shed light on power relationships in space, triggered by the confrontation between global forces on the one hand and local, place-based, or “territorially anchored” groups on the other. The article unfolds across two sections. The first summarizes diverse conceptualizations of space, power, and locality, which are understood to be core elements of territorio, particularly when these are linked through actions, demands, or claims of a collective. The second section approaches the thematic contexts in which territorio and its associated concepts are used: as spatial entities of jurisdictional administration, appropriation by indigenous, afro and peasant communities, political demands and social movements, control-dispossession and reconstruction, spatial entities of assets, and place-based development. We argue that an understanding of the multiple uses and interpretations of territory in Latin America does not just reveal the uniquely nuanced ways of thinking about geography in the region, but it also raises questions over the possibility of a multiple construction of key geographical concepts. Resumen: Este articulo explora los conceptos envueltos en el termino territorio , la naturaleza de los usos del mismo y las interpretaciones dadas por distintos grupos de usuarios del termino en America Latina. Sostenemos que, a pesar de su naturaleza ubicua, su uso polisemico y la hibridez de su construccion conceptual, territorio es un termino especifico latinoamericano, que puede contribuir al analisis de las relaciones de poder en el espacio, activadas por la confrontacion entre fuerzas globales, por un lado, y grupos con “anclaje territorial”, por el otro. El articulo presenta dos secciones. En la primera se explican diversas aproximaciones a los conceptos de espacio, poder y localidad, que son elementos centrales para el entendimiento de territorio, particularmente cuando estos estan conectados a traves de acciones, demandas o reclamos de un colectivo. La segunda seccion aborda los contextos tematicos en los cuales territorio y sus conceptos relacionados son usados: como entidades espaciales de administracion jurisdiccional, apropiacion por grupos indigenas, afros y campesinos, demandas politicas y movimientos sociales, control-desposesion y reconstruccion, como unidades espaciales de recursos y desarrollo basado-en el lugar (o desarrollo territorial). Creemos que el entendimiento de los multiples usos e interpretaciones de “territorio” en America Latina no solo revela los distintivos matices del pensamiento geografico en la region, sino tambien, plantea cuestiones sobre la posibilidad de una construccion multiple de conceptos geograficos fundamentales.

Prior research developed the Attention-Related Driving Errors Scale (ARDES), a 19-item questionnaire aimed at evaluating the drivers’ predisposition to attentional error, and also provided validity evidence of ARDES measures across two countries: Argentina and Spain. In the current work we adapt the Spanish version of the ARDES to the English language and the culture, traffic regulations and driving habits of the UK, and then provide new evidence of the cross-cultural equivalence of the scale. First, an improved forward and backward translation design was used to translate the Spanish ARDES into English. A committee-approach review process was also performed during the translation phase to guarantee that the English ARDES was the most appropriate to reflect the language, culture, traffic regulations and driving habits of the UK. Second, the adapted questionnaire was tested on 301 British drivers to inspect its psychometric properties and study the relationships between the ARDES-UK and different socio-demographic variables. In agreement with the previous literature, the results of the factor analysis confirmed a single factor and accounting for 33.84% of the variance. Cronbach’s alpha was equal to .89. The observed pattern of relationships of the scores and the different socio-demographic variables was consistent with previous evidence and support the validity of the adapted questionnaire. For example, the ARDES-UK scores were significantly different between drivers reporting and not reporting traffic accidents with material damage. To conclude with, the translation design and the resulting psychometric suggest the validity of ARDES-UK measures for evaluating the British drivers’ predisposition to attentional error. In addition, the evidence in the current work supports the hypothesis of the cross-cultural stability of the scale across Spain and the UK.

Prophylaxis with factor VIII concentrates is the cornerstone of treatment for severe hemophilia A. In children, early onset of primary and secondary prophylaxis is the gold standard in most of countries with adequate financial resources. In adults, it is reasonable to continue the prophylactic treatment initiated during childhood to preserve joint functionality. Adults with advanced hemophilic arthropathy can also benefit from tertiary prophylaxis, showing reductions in bleeding episodes, target joints and missed days from work. The long-term evolution of hemophilic patients usually depends on how successfully prophylaxis can prevent bleeding in both children and adults. For prophylaxis to be efficient it is essential to consider the resources available (factor concentrate use and trough levels) and the patient's bleeding phenotype and clinical situation (levels of activity required, presence of chronic synovitis and arthropathy), and to define the annual number of bleeds (particularly into joints) acceptable for each patient.

Diagnosis of von Willebrand disease (VWD) depends on personal and family history of bleeding and confirmatory laboratory testing. Currently available phenotypic tests for VWD contain potential sources for error that may distort results. Despite an exponential growth of information about the von Willebrand factor gene (VWF), the role of molecular diagnosis in VWD is still controversial. Due to the complexity and high cost of conventional molecular analyses, some investigators have recommended limiting this approach to distinguish suspected type 2N VWD from hemophilia A, type 2B from platelet-type VWD, and the exploration of type 3 VWD. New genetic methodologies and approaches are becoming available, but there is still some reluctance for their implementation in VWD diagnosis. This article discusses the pros and cons of molecular testing in VWD considering the experience obtained through the multicenter project “Molecular and Clinical Profile of VWD in Spain (PCM-EVW-ES).”

Dendroctonus bark beetles (Scolytinae) are one of the most important disturbance agents of coniferous forests in North and Central America. These beetles spend their lives almost entirely under the tree bark, and their survival and reproductive success depend on their ability to overcome the toxic effect of the trees’ oleoresin. The cytochromes P450 (CYPs) are associated with the detoxification process of xenobiotics, as well as other physiological processes. Different cytochromes (families 4, 6, and 9) in the Dendroctonus species have been expressed under several experimental conditions; nevertheless, the expression time-course of these genes is unknown. To explore the induction speed of CYPs, we evaluated the relative expression of the CYP6BW5, CYP6DG1, CYP6DJ2, CYP9Z18, and CYP9Z20 genes at the early hours of drilling and settling into a tree (1, 2, 4, 6, 8, 12, 18 h) both in females and males, solitary or paired, of the bark beetle Dendroctonus rhizophagus Thomas and Bright. Our findings show that the five genes were rapidly overexpressed in the early hours (1 to 6 h) in both sexes and in solitary and paired conditions, suggesting their participation in the detoxification process. Additionally, the CYPs expression shows up- and down-regulation patterns through these short times, suggesting their probable participation in other physiological processes as the biosynthesis of hormones, pheromones or compounds related to reproduction.

espanolEl presente articulo analiza la situacion del trabajo femenino en los hogares de Miazi y Shaime, dos comunidades shuar en la Amazonia sur ecuatoriana, en un contexto de colonizacion y de reciente apertura a mercados locales. Se observan ademas las implicaciones del trabajo de las mujeres en la agro-biodiversidad y en la alimentacion de sus familias, asi como la valoracion subjetiva sobre su participacion. Los resultados muestran principalmente dos espacios de produccion agricola en las fincas familiares de los entrevistados: el aja o huerta para autoconsumo y los cultivos de comercializacion. Esta caracterizacion se hace en funcion del rol que las mujeres cumplen en cada espacio y de la carga de trabajo que tienen en el proceso productivo. Ademas, se presentan algunos extractos de entrevistas de varios actores locales y comunitarios que contextualizan las condiciones desiguales y de sobre-explotacion sobre las cuales se desarrolla el trabajo femenino. Este analisis revela que, aunque invisible, el trabajo de las mujeres es central en una economia agricola de mercado en desarrollo en estos ecosistemas ecologicamente sensibles. EnglishThis article analyzes the situation of female work among the households in Miazi and Shaime, two Shuar communities in the Southern Ecuadorian Amazon, in a context of colonization and recent local market opening. The implication of women’s work in terms of agro-biodiversity and their family diet are also observed, as well as the subjective assessment of their participation. Findings depict mainly two spaces of agricultural production in the interviewees’ household farms: the aja or garden for self-consumption and the market crops. This characterization is based on the role that women fulfill in each space and the workload they have in the production process. In addition, some excerpts from interviews to various local and community actors are presented; they contextualize the uneven conditions and over-exploitation on which women’s work develops. This analysis reveals that, although invisible, women’s work is central to a developing agricultural market economy in these ecologically sensitive ecosystems.

We hypothesized that inhibitor specificity may predict the outcome of antifactor VIII autoantibodies eradication treatment in acquired hemophilia A. Our objective was to analyze the association between factor VIII domains recognized by inhibitors and outcome of the immunosuppressive therapies (ISTs) in a prospective, observational study. 16 patients were recruited. Inhibitor specificities were assessed at diagnosis and throughout the study. Their association with IST outcome was addressed. First- line IST succeeded in56% of patients. Inhibitors reacted mainly with light chain domains (69%) and/or the A2 domain (44%). 31% inhibitors recognized more than one domain. Significantly, the number of patients whose inhibitors recognized the light chain was significantly higher in the group of those who did not reach complete remission after first line IST when compared with those who did [6/7 (85.7%) vs. 4/9 (44.4%), P< 0.05]. Therefore, inhibitor specificitycould predict the success of IST in acquired hemophilia A. Copyright (C) 2019 Wolters Kluwer Health, Inc. All rights reserved.

[EN] Vertical/horizontal coordination between different subjects of Chemical Engineering Degree is important in order that students can reach the skills that they will need in a future, in their profession. This means coordinating contents, methodologies and timing of concepts. Therefore, teachers of Campus d’Alcoi have developed a project named “Adsorption column design for the elimination of dyes in wastewater from the textile industry”. The students solve a part of the project using BPL methodology in every subject and complete the project sequentially throughout the degree. This work shows the steps to start this project and an example of the problems proposed in the subject Chemical Engineering Laboratory II., [ES] El nivel de coordinación vertical/horizontal entre las diferentes asignaturas impartidas en los grados es esencial para que el alumno pueda adquirir todas las competencias necesarias para ejercer su profesión. Ello conlleva coordinar contenidos, metodologías y temporalización. En esta línea, los profesores del Campus d’Alcoi han desarrollado un proyecto “Diseño de una columna de adsorción para la eliminación de colorantes en aguas residuales de la industria textil” para que los alumnos lo tengan que resolver parcialmente en cada asignatura mediante la metodología docente de ABP y completarlo de forma secuencial a lo largo de la titulación. En este trabajo se muestra cómo han sido los pasos para poner en marcha el proyecto y un ejemplo de los problemas planteados en la asignatura Experimentación en Ingeniería Química II., Agradecimiento al PIME de la UPV (2017) Referencia A19: “Coordinación en el Grado en Ingeniería Química del Campus d’Alcoi: Aprendizaje Basado en Problemas como Metodología de Conexión entre asignaturas”

Introduction Patients with severe haemophilia and inhibitors against factor VIII who require surgery need a prophylactic approach to prevent bleeding complications. Scientific evidence to decide the best prophylactic treatment is very limited and mainly based on retrospective or case series. Aims To develop evidence- and expert opinion-based guidelines for prophylactic therapy for patients with haemophilia and inhibitors undergoing surgery. Methods A panel of nine Spanish haematologists undertook a systematic review of the literature and selected publications providing relevant information regarding the prophylactic management of patients with haemophilia and inhibitors undergoing dental extraction, minor surgery or major surgery. Results Although evidence is very limited, the panel considers that it seems advisable that prophylaxis should be given in most cases with a bypassing agent (aPCC or rFVIIa) and should start immediately before minor or major surgery. Patients should be closely monitored to enable dose/product modification as needed. Conclusion It is necessary to communicate clinical experience in a detailed way in order to ensure optimal schemes of prophylaxis for patients with haemophilia and inhibitors. Development of objective outcomes to evaluate efficacy is crucial.

SummaryProphylaxis with the blood clotting factor, factor VIII (FVIII) is ineffective for individuals with haemophilia A and high-titre inhibitors to FVIII. Prophylaxis with the FVIII bypassing agents activated prothrombin complex concentrates (aPCC; FEIBA® Baxalta) or recombinant activated factor VII (rFVIIa; Novo-Seven®, Novo Nordisk) may be an effective alternative. It was our aim to develop evidence -and expert opinion- based guidelines for prophylactic therapy for patients with high-titre inhibitors to FVIII. A panel of nine Spanish haematologists undertook a systematic review of the literature to develop consensusbased guidance. Particular consideration was given to prophylaxis in patients prior to undergoing immune tolerance induction (ITI) (a process of continued exposure to FVIII that can restore sensitivity for some patients), during the ITI period and for those not undergoing ITI or for whom ITI had failed. These guidelines offer guidance for clinicians in deciding which patients might benefit from prophylaxis with FVIII bypassing agents, the most appropriate agents in various clinical settings related to ITI, doses and dosing regimens and how best to monitor the efficacy of prophylaxis. The paper includes recommendations on when to interrupt or stop prophylaxis and special safety concerns during prophylaxis. These consensus guidelines offer the most comprehensive evaluation of the clinical evidence base to date and should be of considerable benefit to clinicians facing the challenge of managing patients with severe haemophilia A with high-titre FVIII inhibitors.

In this study, we evaluate landscape composition and configuration changes between 1989 and 2016 associated with the establishment and downsizing of the Corazon de Oro Protective Forest, a mixed-use protected area in the tropical Andes of Southern Ecuador. We identify the most significant land use/land cover changes and the implications for management and conservation linked to these two events. We rely on remote sensing, landscape indicators, statistical analysis, and expert knowledge to evaluate landscape structure. This combination of methods allowed us to quantify diversity, evenness, proportional abundance, dominance, surface area, and isolation trajectories to assess compositional and configurational differences marked by the protected area’s creation and downsizing. While we found that landscape composition remained relatively unchanged over time, the creation of the protected area significantly altered landscape configuration but curbed human pressure on evergreen montane habitats. The downsizing event increased anthropogenic pressure on highland paramo vegetation. Our findings demonstrate that some aspects of landscape configuration vary independently of composition. Most importantly, however, we show that protected area formation and downsizing can significantly alter the spatial arrangement and size of transitional areas, which, in turn, may affect landscape dynamics with implications for energy flow and habitat function. The suite of landscape metrics used in this study are effective indicators that can assist managers monitor the performance of mixed-use protected areas.

[EN] The valorization of olive oil mill solid wastes (OMW) has been addressed by considering it as a possible source of humic-like substances (HLSs), to be used as auxiliary substances for photo-Fenton, employing caffeine as a target pollutant to test the efficiency of this approach. The OMW-HLS isolation encompassed the OMW basic hydrolysis, followed by ultrafiltration and drying. OMW-HLS structural features have been investigated by means of laser light scattering, fluorescence, size exclusion chromatography, and thermogravimetric analysis; moreover, the capability of OMW-HLS to generate reactive species under irradiation has been investigated using spin-trap electronic paramagnetic resonance. The caffeine degradation by means of photo-Fenton process driven at pH = 5 was significantly increased by the addition of 10 mg/L of OMW-HLS. Under the mechanistic point of view, it could be hypothesized that singlet oxygen is not playing a relevant role, whereas other oxidants (mainly OH center dot radicals) can be considered as the key species in promoting caffeine degradation., Authors want to acknowledge the financial support of Spanish Ministerio de Economia y Competitividad (CTQ2015-69832-C04) and European Union (645551-RISE-2014, MAT4-TREAT).

Bark beetles of the genus Dendroctonus are important components of coniferous forests. During host colonization, they must overcome the chemical defences of their host trees, which are metabolized by cytochrome P450 (CYP or P450) enzymes to compounds that are readily excreted. In this study, we report the relative expression (quantitative real-time PCR) of four orthologous cytochrome P450 genes (CYP6BW5, CYP6DG1, CYP6DJ2 and CYP9Z20) in Dendroctonus rhizophagus and Dendroctonus valens forced to attack host trees at 8 and 24 h following forced attack and in four stages during natural colonization [solitary females boring the bark (T1); both male and female members of couples before oviposition (T2); both male and female members of couples during oviposition (T3), and solitary females inside the gallery containing eggs (T4)]. For both species gene expression was different compared with that observed in insects exposed to single monoterpenes in the laboratory, and the expression patterns were significantly different amongst species, sex, gut region and exposure time or natural colonization stage. The induction of genes (CYP6BW5v1, CYP6DJ2v1 and CYP9Z20v1 from D. rhizophagus, as well as CYP6DG1v3 from D. valens) correlated with colonization stage as well as with the increase in oxygenated monoterpenes in the gut of both species throughout the colonization of the host. Our results point to different functions of these orthologous genes in both species.

Resumen Estudio observacional que valoro la seguridad de heparina de bajo peso molecular (HBPM) administrada para la profilaxis o el tratamiento de complicaciones tromboembolicas durante el embarazo. Diseno Se revisaron retrospectivamente historias clinicas y se recogio el resultado del embarazo y los acontecimientos adversos. El riesgo tromboembolico (ETEV) se analizo mediante la clasificacion del Royal College of Obstetrics and Gynaecologist . Resultados Se incluyeron 127 pacientes (131 fetos) con edad media de 32,3 ± 4,3 anos. La HBPM se indico por ETEV aguda en 11 (8,6%) pacientes y por profilaxis en 116 (91,4%). En el grupo de profilaxis hubo 38 (30,0%), 49 (38,6%) y 29 (22,8%) pacientes con riesgo de ETEV alto, moderado y bajo respectivamente. Los nacidos vivos fueron 127 (97%) —19 (15,1%) pretermino—. Una paciente con deficit de antitrombina desarrollo ETEV. Hubo 25 casos de sangrado (18 [72%] hematomas subcutaneos). Conclusion La HBPM es bien tolerada y segura para la profilaxis y el tratamiento de las complicaciones tromboembolicas del embarazo.

The main objective of this prospective observational study is to describe the pharmacokinetic profile of pa- tients with hemophilia A in prophylaxis with Advate® in Spain using myPKFiT®.

The long-Term survival of Andean forest landscapes (AFL) and of their capacity to contribute to sustainable development in a context of global change requires integrated adaptation and mitigation responses informed by a thorough understanding of the dynamic and complex interactions between their ecological and social components. This article proposes a research agenda that can help guide AFL research efforts for the next 15 years. The agenda was developed between July 2015 and June 2016 through a series of workshops in Ecuador, Peru, and Switzerland and involved 48 researchers and development experts working on AFL from different disciplinary perspectives. Based on our review of current research and identification of pressing challenges for the conservation and sustainable governance of AFL, we propose a conceptual framework that draws on sustainability sciences and social-ecological systems research, and we identify a set of high-priority research goals and objectives organized into 3 broad categories: systems knowledge, target knowledge, and transformation knowledge. This paper is intended to be a reference for a broad array of actors engaged in policy, research, and implementation in the Andean region. We hope it will trigger collaborative research initiatives for the continued conservation and sustainable governance of AFL. Fil: Mathez Stiefel, Sarah Lan. University of Bern; Suiza. Centro Internacional de la Papa, Lima; Perú Fil: Peralvo, Manuel. Consortium For The Sustainable Development Of The Andean Ecoregion; Ecuador Fil: Báez, Selene. Consortium For The Sustainable Development Of The Andean Ecoregion; Ecuador. Escuela Politécnica Nacional; Ecuador Fil: Rist, Stephan. University of Bern; Suiza Fil: Buytaert, Wouter. Imperial College London; Reino Unido Fil: Cuesta, Francisco. Institute For Biodiversity And Ecosystem Dynamics; Países Bajos. Consortium For The Sustainable Development Of The Andean Ecoregion; Ecuador Fil: Fadrique, Belén. University of Miami; Estados Unidos Fil: Feeley, Kenneth J.. University of Miami; Estados Unidos Fil: Groth, Aaron A. P.. University of Texas at Austin; Estados Unidos Fil: Homeier, Jürgen. Universität Göttingen; Alemania Fil: Llambí, Luis D.. Universidad de los Andes; Venezuela Fil: Locatelli, Bruno. Centre For International Forestry Research; Perú. Cirad Centre de Recherche de Montpellier; Francia Fil: López Sandoval, Maria Fernanda. Facultad Latinoamericana de Ciencias Sociales; Ecuador Fil: Malizia, Agustina. Universidad Nacional de Tucumán; Argentina. Universidad Nacional de Tucumán. Instituto de Ecología Regional. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Ecología Regional; Argentina Fil: Young, Kenneth R.. University of Texas at Austin; Estados Unidos

Summary Essentials Increasing evidences support a role for splicing defects in multiple disorders. For antithrombin (AT) deficiency only 7% of mutations disturb intronic splicing sequences. Our study of 141 unrelated cases with AT deficiency found higher rate of splicing defects (>13%). A wide range of gene defects cause different types of AT deficiency through aberrant splicing. Background There is increasing evidence supporting the relevance of aberrant splicing in multiple disorders. In antithrombin deficiency only 22 intronic mutations affecting splicing sites (7% of SERPINC1 mutations) are considered as splicing mutations. Methods SERPINC1 was analyzed by Sanger sequencing and MLPA in 141 unrelated cases with antithrombin deficiency. Plasma antithrombin was studied by functional and western blot assays, purified by FPLC and characterized by proteomic analysis. In silico predictions on splicing was done with the Human Splicing Finder software. Results We detected 89 different SERPINC1 defects, 13 with potential effect on splicing. Ten cases presented 9 mutations disturbing splicing sites, 5 new. Three gross or small gene defects also disturbed a correct splicing. Interestingly, the first duplication of a single exon ever described (c.1154-13_1218+115dup), caused mild deficiency (75%). A deeper intronic mutation (c.1154-14G>A), identified in three unrelated patients with traces of disulphide dimers of antithrombin in plasma, created a cryptic splicing site that might generate a variant with 4 additional in frame residues according to in silico predictions. This aberrant splicing was confirmed by proteomic analysis of the dimer purified from plasma. Conclusions A high proportion of cases with antithrombin deficiency (up to 13%) may be explained by an aberrant splicing. Up to 15% of mutations in SERPINC1: splicing site variations, gross gene defects and deep intronic mutations, may affect a correct splicing with three potential consequences type I, type II, and even moderate antithrombin deficiency.

Diagnostics of von Willebrand disease (VWD) includes assessment of factor VIII (FVIII) coagulant activity, von Willebrand factor (VWF) antigen (VWF:Ag) and VWF ristocetin cofactor activity (VWF:RCo), and more specific tests as multimeric and genetic analyses are necessary for the correct VWD classification. The ACL AcuStar analyzer introduces chemiluminescence (CL) technology in detection of VWD with automated VWF:Ag and VWF:RCo assays. Compare VWF:Ag-ELISA and VWF:RCo by aggregometry conventional assays with new CL VWF:Ag-IL and VWF:RCo-IL assays, investigate the ability to make accurate VWD diagnosis and concordance with multimeric and genetic analyses. 146 patients with congenital VWD (51 Type 1; 34 Type 2A; 16 Type 2B; 31 Type 2M; 5 Type 2N; 9 Type 3) and 30 healthy normal subjects were included. A comparison was made between CL and conventional methods. Diagnostic evaluation included: VWF:RCo/VWF:Ag ratio, multimeric distribution (sodium dodecyl sulfate [SDS]-agarose gel) of VWF and genetic analysis in 110 of 146 patients. CL and conventional methods revealed good correlation. Kappa test agreement diagnosis was >0.8. CL diagnostic sensitivity was 100% and specificity 97%. Multimeric and genetic analysis were of help in clarifying 13 discrepancies of diagnosis between methods, of which six discrepancies were explained by lack of conventional methods′ sensibility. CL methodology can detect VWD and discriminate between type 1, 3 and variant forms and offers an automated, faster, sensitive and less cumbersome method when compared to conventional assays, in particular VWF:RCo by aggregometry. In some cases, even with all phenotype and genetic analyses, discrepancies exist in the classification of VWD.

This paper presents how participatory mapping can contribute to the understanding of the local meanings of community limits and perceptions about tenure security in agro-pastoral communities of the Andes, which participate in the payments for environmental services (PES) program. Literature about PES sustains that increasing tenure security might be an additional element of participating in such programs. We focused on the “Socio Paramo” conservation program to analyze how the inclusion of land in this program has infl uenced perception on limits and tenure security for the local communities. With two case studies in the high grasslands (paramo) of Ecuador, we fi rst used sketch maps to elicit how the area inscribed in the PES program has modifi ed the spatial structures of the community territory and what type of limits are found in this territory. Then, we conducted narrative walking to track GPS points with descriptions of land uses, perceived communitarian limits and narratives about meaning or concerns with regard to limits for the community and the relation between the legalized area inscribed in the PES program and tenure security. Maps were produced through GIS support and narratives were analyzed through thematic coding. The study reveals that: a) legal tenure obtained in frame of PES implementation is perceived as a pre-condition to participate in the program and has infl uenced positively or negatively land tenure security for the entire community; b) knowledge about community limits of paramo is the main concern for elder members, who regard this knowledge as the key element to maintain community cohesion. We conclude that participatory mapping is a powerful tool to elucidate concerns about limits, rules and control over land use and persistence of communitarian life, elements that should be considered when implementing PES.

The Attention-Related Driving Errors Scale (ARDES) is a self-reported questionnaire to assess individual differences in the proneness to make attentional errors while driving. The aims of the current work are to adapt the original Argentinean version of the ARDES to the culture, language, traffic regulations and driving habits of Spain and provide new validity evidence of the cross-cultural equivalence of the scale. In the first step of the validation process, five external independent experts reviewed the original ARDES-Argentina and proposed modifications, adapted to the culture, language, traffic regulations and driving habits in Spain. Secondly, a sample of 320 drivers completed the adapted questionnaire. Descriptive statistics and psychometric properties (corrected item-total correlation, Cronbach’s alpha and factor structure) were performed on the data. Besides, in order to obtain further validity evidence, the relationships between the questionnaire scores and socio-demographic variables (age, sex, educational level, driving experience, crash involvement and traffic fines received) were analyzed. Factor analysis suggested a single factor that exceeded the parallel analysis criterion and accounted for 32.70% of the total variance. All items showed positive loadings on this factor, ranging from .41 to .72. The corrected item-total correlation values extend from .41 to .60, indicating that the items had good discrimination power. Cronbach’s alpha coefficient value was .88. The analysis of the relationships between ARDES-Spain scores and socio-demographic variables provided further validity evidence of the appropriateness of the adapted questionnaire. In particular, differences in ARDES-Spain scores were found between drivers who reported traffic collisions with material damage and participants who did not. In conclusion, results in the current study suggest that the adapted version of the ARDES is a useful tool for evaluating the proneness to attentional errors during driving in the Spanish population. Future studies adapting the questionnaire to other countries with different languages, cultures, traffic regulations and driving habits are encouraged in order to expand the discussion on the cross-cultural equivalence of the ARDES.

Forest remnants of La Carbonería are the last patches of natural vegetation near the city of Ibarra. These forest patches are now isolated and restricted to areas of rugged topography and sustain a scarcely explored, largely unknown flora. As an example of this, two new species of orchids were discovered in our first exploration: Lepanthes ibarrae and Lepanthes obandoi, which are described and illustrated in this article. Lepanthes ibarrae is similar to L. zapatae , but is distinguished by its arched stems, reduced petals with a small, rounded apicule, lip with lanceolate-oblong, papillose laminae, small, rounded appendix covered with white pubescence at apex and the triangular, long-attenuate, reflexed rostelum. Lepanthes obandoi is reminiscent of L. muscula, L. guanacasensisr & R. Escobar and L. osiris, but can be distinguished by the much taller plant with distinct superposed stems, elongate appendix with orbicular, pubescent apex and a prominent tuft of white hairs at the base.

Bark beetles (Curculionidae: Scolytinae) are major cause of woody plants death in the world. They colonize the stem and other parts of trees recognizing host-produced specific compounds (kairomones) and insect pheromones. Bark beetle's antennae and alimentary canal participate in the host selection identifying chemical compounds produced by trees and insects, and also in the metabolism and detoxification of these compounds. The red turpentine beetle (RTB), Dendroctonus valens LeConte, is an unaggressive species that colonize > 40 pine species (Pinaceae) in North and Central America. Several studies suggest that bark beetle cytochrome P450 enzymes are involved in monoterpene oxidation. In this study we identified by means of PCR, cloning, sequencing, and bioinformatic analysis, eleven full-length genes: five CYP4, four CYP6, and two CYP9 in the antennae and gut region of RTB, after stimulation with vapors of monoterpenes: (±)-α-pinene, (R)-(+)-α-pinene, (S)-(−)-β-pinene, (S)-(−)-α-pinene and (+)-3-carene; pine trees volatiles used by RTB as kairomones. The recovered cDNA of these genes vary from 1.5 kb to 1.8 kb and the open frame encodes from 496 to 562 amino acid proteins. The bioinformatic analysis suggests that the majority of P450 proteins encoded by these genes are membrane anchored in the endoplasmic reticulum. RT-qPCR assays showed differential expression of all CYP genes between male and female. The gene expression was dependent of monoterpenes and exposure time, with some of them sex, antennae and gut region specific. Significant differences among monoterpenes, gut region, antennae and exposure time were found. Our results suggest that some of these genes may be involved in the detoxification process of these compounds during tree colonization.

We identified the species of bats and their roosts in the urban area of Lules, Tucuman. This is the first systematic study in Argentina in an urban area. Seven species were collected mainly housed in roofs, outstanding the collection of Eumops glaucinus, representing the second record of this species for the province.

Driver distraction and inattention are considered among the major contributing factors in road traffic crashes. One of the most widely used tools to study drivers’ attentional lapses and other types of aberrant behaviour is the Driver Behaviour Questionnaire (DBQ). In the present work, further evidence of the feasibility of the DBQ to study driver inattention is provided. The relationships between the DBQ and both a computer-based neurocognitive test on attentional performance (the Attention Network Test for Interactions and Vigilance, ANTI-V) and a self-reported measure of cognitive failure (the Cognitive Failures Questionnaire, CFQ) are analysed. Results show that attentional lapses are negatively associated with vigilance and positively associated with cognitive failure. Other types of aberrant behaviour (driving errors, traffic violations and aggressive behaviours) were not found related to the attentional performance indices (executive control, orienting, phasic alertness or vigilance), with the exception of DBQ-Violations and the executive control score in percentage of errors. In addition, the relationship between the other types of aberrant behaviour with cognitive failure was more moderate (except for DBQ-Errors, which was also highly correlated). Overall, these results are consistent with the idea of DBQ-Lapses being related to driving inattention, and suggest that this subscale could be a useful tool to study vigilance-related driving behaviour. Further evidence with improved versions of the DBQ or alternative questionnaires would be helpful to clarify whether proneness to attentional lapses while driving may be associated with different driving performance measures, such as crashes or near misses.

Biochemical effects of azinphosmethyl (AZM), an organophosphate pesticide, were determined in gill, brain and muscle tissues of Odontesthes hatcheri and Jenynsia multidentata. The 96-h toxicity was first assessed, estimating lethal concentrations fifty (LC50) of 7 and 30 μg L−1 AZM for O. hatcheri and J. multidentata, respectively. Considering the LC50, sublethal 96-h static exposures were designed for O. hatcheri (0.1–0.5 μg L−1 AZM) and J. multidentata (5–10 μg L−1AZM) to determine biochemical endpoints. Brain acetylcholinesterase (AchE) was inhibited by AZM in both species, while the buffer enzyme carboxylesterase (CarbE) was not affected in this tissue. Conversely, muscular AchE was not affected but CarbE was augmented by AZM. The enzymes glutathione reductase, glutathione-S-transferase and CarbE were significantly inhibited in O. hatcheri gills but none of them was affected by AZM in J. multidentata gills compared to control. GSH levels were augmented in gills of both species in exposed fish compared to controls and in addition, lipid peroxidation was significantly increased in O. hatcheri gills. Ex vivo histochemical analysis of ROS by fluorescence microscopy was also performed in J. multidentata gills, indicating a significant increase upon exposure to 10 μg L−1 AZM. Principal component analyses (PCA) were applied, both to the species together or separately. The general analysis demonstrated a clear separation of responses in the two species. For O. hatcheri, the variable that explains the major variation in PC1 is gill catalase and brain AchE in PC2. In J. multidentata in turn, the variable that explains the major variation in PC1 is brain AchE and total oxyradical scavenging capacity in PC2. The toxicity data and biomarker responses obtained for both species were compared to environmental concentrations of AZM detected in superficial water from different points in the Alto Valle region and risk quotients (RQ) were calculated. This approach indicated probable acute effects for O. hatcheri in river and irrigation channels (RQ > 0.1), while the risk was unacceptable in drainage superficial water (RQ > 1). In contrast, J. multidentata showed minimal risk in river or channel water (RQ

SummaryThe diagnosis of von Willebrand disease (VWD) remains difficult in a significant proportion of patients. A Spanish multicentre study investigated a cohort of 556 patients from 330 families who were analysed centrally. VWD was confirmed in 480. Next generation sequencing (NGS) of the whole coding VWF was carried out in all recruited patients, compared with the phenotype, and a final diagnosis established. A total of 238 different VWF mutations were found, 154 were not included in the Leiden Open Variation Database (LOVD). Of the patients, 463 were found to have VWF mutation/s. A good phenotypic/ genotypic association was estimated in 96.5 % of the patients. One hundred seventy-four patients had two or more mutations. Occasionally a predominant phenotype masked the presence of a second abnormality. One hundred sixteen patients presented with mutations that had previously been associated with increased von Willebrand factor (VWF) clearance. RIPA unavailability, central phenotypic results disagreement and difficult distinction between severe type 1 and type 3 VWD prevented a clear diagnosis in 70 patients. The NGS study facilitated an appropriate classification in 63 of them. The remaining seven patients presented with a VWF novel mutation pending further investigation. In five patients with a type 3 and two with a type 2A or 2B phenotype with no mutation, an acquired von Willebrand syndrome (AVWS) was suspected/confirmed. These data seem to support NGS as a first line efficient and faster paradigm in VWD diagnosis.Supplementary Material to this article is available online at www.thrombosis-online.com.

SummaryAntithrombin is the main endogenous anticoagulant. Impaired function or deficiency of this molecule significantly increases the risk of thrombosis. We studied the genetic variability of SERPINC1, the gene encoding antithrombin, to identify mutations affecting regulatory regions with functional effect on its levels. We sequenced 15,375 bp of this gene, including the potential promoter region, in three groups of subjects: five healthy subjects with antithrombin levels in the lowest (75%) and highest (115%) ranges of our population, 14 patients with venous thrombosis and a moderate antithrombin deficiency as the single thrombophilic defect, and two families with type I antithrombin deficiency who had neither mutations affecting exons or flanking regions, nor gross gene deletions. Our study confirmed the low genetic variability of SERPINC1, particularly in the coding region, and its minor influence in the heterogeneity of antithrombin levels. Interestingly, in one family, we identified a g.2143 C>G transversion, located 170 bp upstream from the translation initiation codon. This mutation affected one of the four regions located in the minimal promoter that have potential regulatory activity according to previous DNase footprinting protection assays. Genotype-phenotype analysis in the affected family and reporter analysis in different hepatic cell lines demonstrated that this mutation significantly impaired, although it did not abolish, the downstream transcription. Therefore, this is the first mutation affecting a regulatory region of the SERPINC1 gene associated with antithrombin deficiency. Our results strongly sustain the inclusion of the promoter region of SERPINC1 in the molecular analysis of patients with antithrombin deficiency.

Based on Posner's (1994) model of attentional functions, the relationship between age and personal proneness to attention-related errors while driving and the functioning of the 3 attentional networks were explored by means of attentional behavioral measures and self-report data.A sample of 55 drivers was drawn from the general population of Mar del Plata, Buenos Aires, Argentina. The Attention Network Test for Interactions (ANTI) (Callejas et al. 2004) task was used to obtain behavioral measures of the attentional networks, and we used the Attention-Related Driving Error Scale (ARDES) (Ledesma et al. 2010) questionnaire to obtain a self-report measure of attention-related driving errors.Drivers reporting the greatest propensity to experience attention-related errors showed an overall slowdown in performance, less endogenous preparation for high = priority warning signs, and a better response to conflict in the presence of valid cues than drivers less prone to attention-related errors while driving. Older participants showed a slowdown in performance, less endogenous preparation for high-priority warning signs, and worse cognitive control when solving conflict in comparison with younger drivers.We suggest that each group variable, attention-related error proneness and age, shows a particular combination of attentional network functioning that implies different ways of being distracted, which have different practical implications for safe driving. It can be inferred that drivers who are more prone to commit attentional errors while driving run less risk in situations in which they can deal with response conflict in the presence of valid cues because of the particular way in which their attentional networks are combined when a valid cue is present, could serve to compensate, with a better response conflict, their general slowdown and less endogenous preparedness for high-priority signs. It can be inferred that older drivers might show a reduction of the general state of alertness to signs that indicate the presence of a dangerous driving situation and in solving conflict in traffic circumstances.

We used several cue–target SOAs (100, 500, 1000 ms) and three different degrees of cue predictability (Non-predictive-50%, Predictive-75%, Counter-predictive-25%), to investigate the role of awareness of cue–target predictability on cueing effects. A group of participants received instructions about the informative value of the cue, while another group did not receive such instructions. Participants were able to extract the predictive value of a spatially peripheral cue and use it to orient attention, whether or not specific instructions about the predictive value of the cue were given, and no matter their ability to correctly report it in a post-test questionnaire. In the non-predictive block, bad estimators who received no instructions showed regular cueing effects, while good estimators exhibited smaller and non-significant facilitatory effects at the short SOA and an absence of significant IOR at longer SOAs. However, for the instructions group, the pattern of results reversed.

Bark beetles (Coleoptera: Curculionidae: Scolytinae) are economically and ecologically the most important group of forest insects. They use several monoterpenes produced by their host plants to locate and colonize trees. Some of these compounds also are metabolized to produce sex, aggregation, or antiaggregation pheromones. Experimental studies have shown that certain terpenes are highly toxic to the insects, negatively affecting their development, reproductive success, and colonization of host trees. Nevertheless, the effects of these compounds on specific organs or anatomical systems are unknown. Based on relationship between bark beetles and monoterpenes of their host trees and the midgut morphological characterization performed previously, we studied the effects of α-pinene and myrcene on midgut cells of the red turpentine beetle, Dendroctonus Valens LeConte. Our results show that both α-pinene and myrcene induce an increase in the numbers of lysosomes and mitochondria. Outer and inner mitochondrial membranes were very conspicuous but were not disrupted. Both smooth and rough endoplasmic reticula were abundant and were found throughout the cytoplasm. Two kinds of secretory vesicles were observed: one associated with digestive material and the other associated with “flocculent” material. The Golgi complex was prominent but had no defined arrangement. Observed ultrastructural changes are indicative of intense cell activity and are associated with processes of digestion, synthesis, and excretion as well as of monoterpenes transformation but are not indicative of irreversible cellular damage or death.

El trastorno por déficit de atención e hiperactividad (TDAH) afecta a entre un 5% y un 10% de la población en desarrollo. Hiperactividad, impulsividad e inatención son los síntomas comportamentales que mejor caracterizan a este trastorno. A pesar de la extensa investigación clínica en relación al mismo, el déficit subyacente sigue aún generando bastante controversia. Por ello, estudiar las redes atencionales en relación a la caracterización sintomática del TDAH puede ayudar a tener una visión más clara del trastorno y con ello generar mejoras en su tratamiento.

Las tecnicas de registro y analisis tridimensional se han desarrollado de manera importante en los ultimos anos, al grado de constituirse en poderosas herramientas de apoyo para la preservacion del patrimonio cultural. Esta investigacion presenta el Laboratorio de Documentacion y Analisis Tridimensional (Lab3D) de la Coordinacion Nacional del Conservacion del Patrimonio Cultural (CNCPC), Instituto Nacional de Antropologia e Historia (INAH), mediante el estudio de caso del registro tridimensional del conjunto de relieves decorativos en piedra de la zona arqueologica de Tula, Hidalgo, Mexico, el cual servira para el monitoreo y la toma de decisiones en materia de conservacion de este importante legado patrimonial.

The correct diagnosis and classification of von Willebrand disease (VWD) is difficult because of the variability of its clinical expression and limitations of laboratory methods. However, correctly diagnosing VWD is important for therapy and genetic counselling. A survey related to the referred VWD patients in Spain revealed local diagnostic problems in at least one third of the cases of VWD. Consequently, a Spanish multicenter study was carried out in which a cohort of 556 patients from 330 families was analyzed centrally. VWD was confirmed in 480 patients. Next generation sequencing (NGS) of the whole coding von Willebrand factor gene ( VWF ) was carried out in all recruited patients, compared with the phenotype, and a final diagnosis was established. A total of 238 different VWF mutations were found: 154 were not included in the Leiden Open Variation Database (LOVD). Of the patients, 467 were found to have a VWF mutation/s. A good phenotype/genotype association was estimated in 94.6% of the cases. One hundred and eighty patients had two or more mutations. Occasionally a predominant phenotype masked the presence of a second abnormality. One hundred and sixteen patients presented with mutations that had previously been associated with an increased VWF clearance. Ristocetin induced platelet agglutination (RIPA) unavailability, central phenotypic results disagreement and difficult distinction between severe type 1 and type 3 VWD prevented a clear diagnosis in 74 patients. A local/central disagreement over diagnosis occurred in 42.3% of the whole cohort. The NGS study facilitated an appropriate classification in 60 of them. Based on these results, a proposal for a change in the VWD diagnostic paradigm, suggesting the inclusion of the analysis of VWF at the initial diagnostic process, has been made. An extension of this project to include 400 new VWD patients has begun. A diagnostic strategy has been developed, and a Spanish consensus guideline for optimal treatment of VWD was developed in Spain. Desmopressin (DDAVP) is the treatment of choice in responsive VWD patients. VWF concentrates (VWF/FVIII) are used in individuals unresponsive to DDAVP, when DDAVP is contraindicated, or in VWD types 2B and 3, and for long term management needs (e.g., post major surgery).

Objective: The aim of this research was (a) to study driver inattention as a trait-like variable and (b) to provide new evidence of validity for the Attention-Related Driving Errors Scale (ARDES). Background: Driving inattention is approached from an individual differences perspective. We are interested in how drivers vary in their propensity to experience failures of attention and in the methods to measure these differences. Method: In a first sample (n = 301), we tested, via confirmatory factor analysis, a new theoretical model for the ARDES. In a second sample (n = 201), we evaluated the relationship between inattention and internal and external sources of distraction and social desirability bias in ARDES responses. A subsample (n = 65) was reevaluated to study temporal stability of the ARDES scores. Results: Errors measured by the ARDES can be classified according to the driving task level at which they occur (navigation, maneuvering, or control). Differences in ARDES scores based on collision history were observed. ARDES was related to internal sources of distraction and was independent of the level of exposure to distracting activities. Test-retest showed a high degree of stability in ARDES scores. Low correlations were found with a social desirability measure. Conclusion: ARDES appears to measure a personal trait that remains relatively stable over time and is relatively independent of distracting activities. New evidence of validity emerged for this self-report. Application: ARDES can be used to measure individual differences in driving inattention and to help tailor preventive interventions for inattentive drivers. It can serve as an instrument of driver self-assessment in educational and training contexts. Fil: Ledesma, Ruben Daniel. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Montes, Silvana Andrea. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Poó, Fernando Martín. Universidad Nacional de Mar del Plata; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: López Ramón, María Fernanda. Universidad de Zaragoza; España

Glycosyltransferases play diverse roles in cellular metabolism by modifying the activities of regulatory metabolites. Three stress-regulated UDP-glucosyltransferase-encoding genes have been isolated fromthe stigmas of saffron, UGT85U1, UGT85U2 and UGT85V1, which belong to the UGT85 family that includes members associated with stress responses and cell cycle regulation. Arabidopsis constitutively expressing UGT85U1 exhibited and increased anchor root development. No differences were observed in the timing of root emergence, in leaf, stem and flower morphology or flowering time. However, salt and oxidative stress tolerance was enhanced in these plants. Levels of glycosylated compounds were measured in these plants and showed changes in the composition of several indole-derivatives. Moreover, auxin levels in the roots were higher compared to wild type. The expression of several key genes related to root development and auxin homeostasis, including CDKB2.1, CDKB2.2, PIN2, 3 and 4; TIR1, SHR, and CYCD6, were differentially regulated with an increase of expression level of SHR, CYCD6, CDKB2.1 and PIN2. The obtained results showed that UGT85U1 takes part in root growth regulation via auxin signal alteration and the modified expression of cell cycle-related genes, resulting in significantly improved survival during oxidative and salt stress treatments. Spanish Ministerio de Economia y Competitividad BIO2013-44239-R IBERCAROT network 112RT0445 FPCYTA through the INCRECYT Programme

Isolated microspores are reprogrammed in vitro by stress, becoming totipotent cells and producing embryos and plants via a process known as microspore embryogenesis. Despite the abundance of data on auxin involvement in plant development and embryogenesis, no data are available regarding the dynamics of auxin concentration, cellular localization and the expression of biosynthesis genes during microspore embryogenesis. This work involved the analysis of auxin concentration and cellular accumulation; expression of TAA1 and NIT2 encoding enzymes of two auxin biosynthetic pathways; expression of the PIN1-like efflux carrier; and the effects of inhibition of auxin transport and action by N-1-naphthylphthalamic acid (NPA) and α-(p-chlorophenoxy) isobutyric acid (PCIB) during Brassica napus microspore embryogenesis. The results indicated de novo auxin synthesis after stress-induced microspore reprogramming and embryogenesis initiation, accompanying the first cell divisions. The progressive increase of auxin concentration during progression of embryogenesis correlated with the expression patterns of TAA1 and NIT2 genes of auxin biosynthetic pathways. Auxin was evenly distributed in early embryos, whereas in heart/torpedo embryos auxin was accumulated in apical and basal embryo regions. Auxin efflux carrier PIN1-like gene expression was induced in early multicellular embryos and increased at the globular/torpedo embryo stages. Inhibition of polar auxin transport (PAT) and action, by NPA and PCIB, impaired embryo development, indicating that PAT and auxin action are required for microspore embryo progression. NPA also modified auxin embryo accumulation patterns. These findings indicate that endogenous auxin biosynthesis, action and polar transport are required in stress-induced microspore reprogramming, embryogenesis initiation and progression., This work was supported by the Spanish Ministry of Economy and Competitiveness (MINECO) [projects BFU2008-00203, BFU2011-23752 and AGL2014-52028-R; and a pre-doctoral FPI fellowship (BES-2009-014245) to H.R.S.]; and the European Regional Development Fund (ERDF/FEDER) of the European Commission.