Affinity partitioning of proteins tagged with choline-binding modules in aqueous two-phase systems

We present a novel procedure for affinity partitioning of recombinant proteins fused to the cholinebinding module C-LytA in aqueous two-phase systems containing poly(ethylene glycol) (PEG). Proteins tagged with the C-LytA module and exposed to the two-phase systems are quantitatively localized in the PEG-rich phase, whereas subsequent addition of the natural ligand choline specifically shifts their localization to the PEG-poor phase by displacement of the polymer from the binding sites. The described procedure is simple, scalable and reproducible, and has been successfully applied to the purification of four diverse proteins, resulting in high yields and purity

Satélite de altar resolución espacial en la determinación de la fracción de cabida cubierta en sistemas adehesados

La dehesa es un ecosistema de creación humana a partir del bosque de encinas primitivas en el que se han aclarando los árboles y controlado el estrato arbustivo para que se estabilice el pastizal. Su densidad arbórea es muy variable y depende del tratamiento recibido a lo largo de los años. La obtención de la densidad de cobertura arbórea de dicho sistema se calcula mediante la Fracción de Cabida Cubierta (FCC), medida que relaciona la superficie cubierta por la copa del árbol respecto del resto de superficie. En comparación a las imágenes utilizadas actualmente para la obtención de la FCC como ortofotografías o vuelos fotogramétricos, las imágenes satélite de alta resolución espacial ofrecen suficiente detalle espacial para poder aislar la copa de los árboles del entorno, unido a la multiespectralidad de los datos que permite su correcta caracterización espectral. El objetivo de este trabajo fue desarrollar una metodología con imágenes pan-sharpened QuickBird que, a partir del análisis espectral de los árboles, generó imágenes en formato binario árbol-no árbol mediante técnicas de combinación de bandas para la obtención de la FCC. Los resultados obtenidos fueron satisfactorios al obtener fiabilidades mayores del 90% en la determinación de las copas de los árboles en la mayoría de los análisis realizados.The dehesa is an ecosystem developed from primitive oak forest where trees have been clarified and the shrub layer controlled to stabilize the pasture. The tree density is very variable and depends on the treatment received over the years. This density can be calculated by the Canopy Cover Fraction (CCF), a measure that relates the area covered by the trees over the other surface. Satellite imagery of high spatial resolution provide enough spatial detail to isolate the treetops from the environment, and enough multispectral data to allow the correct spectral characterization. The aim of this study was to develop a methodology with QuickBird pan-sharpened imagery for obtaining the CCF based on a spectral analysis of trees and a binary reclassification (tree- no tree) of each band. The results were satisfactory with accuracies greater than 90% in the discrimination of trees in most of the analysis

Determinants of HDL Cholesterol Efflux Capacity after Virgin Olive Oil Ingestion: Interrelationships with Fluidity of HDL Monolayer

Scope: Cholesterol efflux capacity of HDL (CEC) is inversely associated with cardiovascular risk. HDL composition, fluidity, oxidation, and size are related with CEC. We aimed to assess which HDL parameters were CEC determinants after virgin olive oil (VOO) ingestion. Methods and results: Post‐hoc analyses from the VOHF study, a crossover intervention with three types of VOO. We assessed the relationship of 3‐week changes in HDL‐related variables after intervention periods with independence of the type of VOO. After univariate analyses, mixed linear models were fitted with variables related with CEC and fluidity. Fluidity and Apolipoprotein (Apo)A‐I content in HDL was directly associated, and HDL oxidative status inversely, with CEC. A reduction in free cholesterol, an increase in triglycerides in HDL, and a decrease in small HDL particle number or an increase in HDL mean size, were associated to HDL fluidity. Conclusions: HDL fluidity, ApoA‐I concentration, and oxidative status are major determinants for CEC after VOO. The impact on CEC of changes in free cholesterol and triglycerides in HDL, and those of small HDL or HDL mean size, could be mechanistically linked through HDL fluidity. Our work points out novel therapeutic targets to improve HDL functionality in humans through nutritional or pharmacological interventions.Fil: Fernández Castillejo, Sara. Universitat Rovira I Virgili; EspañaFil: Rubió, Laura. Universitat Rovira I Virgili; España. Universidad de Lleida; EspañaFil: Hernáez, Álvaro. Centro de Investigación Biomédica en Red Fisiopatología de la Obesidad y Nutrición; EspañaFil: Catalán, Úrsula. Universitat Rovira I Virgili; EspañaFil: Pedret, Anna. Universitat Rovira I Virgili; EspañaFil: Valls, Rosa M.. Universitat Rovira I Virgili; EspañaFil: Mosse, Juana Inés. Universidad de Lleida; España. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Covas, Maria Isabel. Centro de Investigación Biomédica en Red Fisiopatología de la Obesidad y Nutrición; EspañaFil: Remaley, Alan T.. National Institutes of Health; Estados UnidosFil: Castañer, Olga. Centro de Investigación Biomédica en Red Fisiopatología de la Obesidad y Nutrición; EspañaFil: Motilva, Maria José. Universidad de Lleida; EspañaFil: Solá, Rosa. Universitat Rovira I Virgili; Españ

Assessment of the Persistence of Avena sterilis L. Patches in Wheat Fields for Site-Specific Sustainable Management

This paper aims to evaluate the spatial persistence of wild oat patches in four wheat fields over time to determine the economic feasibility of using late-season wild oat maps for early site-specific weed management (SSWM) next season. The spatial persistence of wild oat patches was analyzed by three tests: land use change detection between years, spatial autocorrelation, and análisis of spreading distance. The temporal trend of wild oat patch distribution showed a clear persistence and a generalized increase in the infested area, with a noticeable level of weed aggregation and a tendency in the new weed patches to emerge close to older ones. To economically evaluate the SSWM, five simulations in four agronomic scenarios, varying wheat yields and losses due to wild oat, were conducted. When yield losses due to wild oat were minimal and for any of the expected wheat yields, some SSWM simulations were more economically profitable than the overall application in most of the fields. Nevertheless, when the yield losses due to wild oat were maximal, all SSWM simulations were less profitable than overall treatment in all the analyzed fields. Although the economic profit variations achieved with SSWM treatments were modest, any of the site-specific treatments tested are preferred to herbicide broadcast over the entire field, in order to reduce herbicide and environmental pollution

Aplicación racional de los nuevos criterios de la European Society for Paediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN) 2020 para el diagnóstico de la enfermedad celíaca

Coeliac disease is a systemic immune-mediated disorder triggered by the ingestion of gluten, which is given in genetically predisposed subjects. It manifests with a wide variety of clinical symptoms, specific serological markers, HLA-DQ2/DQ8 haplotype, and enteropathy. The criteria followed for this have usually been those established by the European Society for Paediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN) since 1969. These criteria have advanced from the need of several intestinal biopsies to, thanks to the development of serological tests of high sensitivity and specificity, considering the enteropathy as one more element in this diagnosis and makes it possible to perform a diagnosis without the need of an intestinal biopsy in certain circumstances. The updated review of the 2012 criteria in 2019 provides new evidence on some aspects, such as the role of HLA, the diagnosis of asymptomatic patients, and the effectiveness of the serological markers. These aspects are reviewed in detail, with the aim of facilitating the rational application of the new 2020 criteria at all care levels. In this sense, Paediatric Primary Care is fundamental in the search for active cases and to perform a first serological study, being recommended that the diagnosis is always establishedby a Paediatric GastroenterologistLa enfermedad celíaca es un proceso sistémico de carácter inmunológico, desen-cadenado por el consumo de gluten, que se da en sujetos genéticamente predispuestos. Se expresa con una gran variedad de síntomas clínicos, marcadores serológicos específicos, hap-lotipo HLA-DQ2/DQ8 y enteropatía. El tratamiento consiste en eliminar de por vida el gluten de la dieta, por lo que es fundamental un diagnóstico adecuado. Los criterios seguidos para ello han sido habitualmente los establecidos por la European Society for Paediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN) desde 1969. Estos criterios han ido evolucionando desde la necesidad de varias biopsias intestinales para el diagnóstico a, gracias al desarrollo de pruebas serológicas de alta sensibilidad y especificidad, considerar la enteropatía como un elemento más en este diagnóstico y posibilitar en determinadas circunstancias realizarlo sin necesidad de biopsia intestinal. La revisión actualizada en 2019 de los criterios 2012 aporta nueva evidencia sobre algunos aspectos, como el papel del HLA, el diagnóstico de los pacientes asintomáticos y la eficacia de los marcadores serológicos. Estos aspectos se revisan en detalle,con el objetivo de facilitar la aplicación de los nuevos criterios 2020 de una forma racional en todos los niveles asistenciales. En este sentido el pediatra de Atención Primaria es fundamental para la búsqueda activa de casos y realizar un primer estudio serológico, recomendándose que el diagnóstico sea siempre establecido por un pediatra gastroenterólog

A Multi-Omics Analysis Pipeline for the Metabolic Pathway Reconstruction in the Orphan Species Quercus ilex

Holm oak (Quercus ilex) is the most important and representative species of the Mediterranean forest and of the Spanish agrosilvo-pastoral “dehesa” ecosystem. Despite its environmental and economic interest, Holm oak is an orphan species whose biology is very little known, especially at the molecular level. In order to increase the knowledge on the chemical composition and metabolism of this tree species, the employment of a holistic and multi-omics approach, in the Systems Biology direction would be necessary. However, for orphan and recalcitrant plant species, specific analytical and bioinformatics tools have to be developed in order to obtain adequate quality and data-density before to coping with the study of its biology. By using a plant sample consisting of a pool generated by mixing equal amounts of homogenized tissue from acorn embryo, leaves, and roots, protocols for transcriptome (NGS-Illumina), proteome (shotgun LC-MS/MS), and metabolome (GC-MS) studies have been optimized. These analyses resulted in the identification of around 62629 transcripts, 2380 protein species, and 62 metabolites. Data are compared with those reported for model plant species, whose genome has been sequenced and is well annotated, including Arabidopsis, japonica rice, poplar, and eucalyptus. RNA and protein sequencing favored each other, increasing the number and confidence of the proteins identified and correcting erroneous RNA sequences. The integration of the large amount of data reported using bioinformatics tools allows the Holm oak metabolic network to be partially reconstructed: from the 127 metabolic pathways reported in KEGG pathway database, 123 metabolic pathways can be visualized when using the described methodology. They included: carbohydrate and energy metabolism, amino acid metabolism, lipid metabolism, nucleotide metabolism, and biosynthesis of secondary metabolites. The TCA cycle was the pathway most represented with 5 out of 10 metabolites, 6 out of 8 protein enzymes, and 8 out of 8 enzyme transcripts. On the other hand, gaps, missed pathways, included metabolism of terpenoids and polyketides and lipid metabolism. The multi-omics resource generated in this work will set the basis for ongoing and future studies, bringing the Holm oak closer to model species, to obtain a better understanding of the molecular mechanisms underlying phenotypes of interest (productive, tolerant to environmental cues, nutraceutical value) and to select elite genotypes to be used in restoration and reforestation programs, especially in a future climate change scenario

Soil mapping based on landscape classification in the semiarid Chaco, Argentina

The semiarid Chaco shared with Argentina, Paraguay, Bolivia, and Brazil, is an ecosystem where the change of land use from forest to commercial agriculture and social conflicts have been intense since the beginning of the 21st century. These changes and the lack of reliable soil information at appropriate scales threaten the sustainable development of the region. In Santiago del Estero province, Argentina, a soil survey was conducted in order to reduce the knowledge gap. Due to the large area, geomorphological diversity, limited funding, and high information demand, a geopedological survey using remote sensing and GIS was considered an suitable approach. The cartographic units were determined based on the integration of geoforms and soils, knowledge of landscape and soil forming factors, field observations, and laboratory determinations. Three main landscape units were recognized: 1) the Chaco fluvio-eolian plain, including a megafan with Haplustolls and Torripsamments, 2) the Rio Dulce valley with Torripsamments, and 3) the migratory alluvial plain of Río Salado with Torripsamments, Ustifluvents, and Natraqualfs. The approach used helped accelerate the collection of soil information at an appropriate scale for land use planning.Sociedad Argentina de Informática e Investigación Operativa (SADIO

Hereditary Leiomyomatosis and Renal Cell Cancer Syndrome in Spain: Clinical and Genetic Characterization

Simple Summary Hereditary leiomyomatosis and renal cell cancer (HLRCC) syndrome is a very rare hereditary disorder characterized by cutaneous leiomyomas (CLMs), uterine leiomyomas (ULMs), renal cysts (RCys) and renal cell cancer (RCC), with no data on its prevalence worldwide. No genotype-phenotype associations have been described. The aim of our study was to describe the genotypic and phenotypic features of the largest series of patients with HLRCC from Spain reported to date. Of 27 FH germline pathogenic variants, 12 were not previously reported in databases. Patients with missense pathogenic variants showed higher frequencies of CLMs, ULMs and RCys, than those with loss-of-function variants. The frequency of RCCs (10.9%) was lower than those reported in the previously published series. Hereditary leiomyomatosis and renal cell cancer syndrome (HLRCC) is a very rare hereditary disorder characterized by cutaneous leiomyomas (CLMs), uterine leiomyomas (ULMs), renal cysts (RCys) and renal cell cancers (RCCs). We aimed to describe the genetics, clinical features and potential genotype-phenotype associations in the largest cohort of fumarate hydratase enzyme mutation carriers known from Spain using a multicentre, retrospective study of individuals with a genetic or clinical diagnosis of HLRCC. We collected clinical information from medical records, analysed genetic variants and looked for genotype-phenotype associations. Analyses were performed using R 3.6.0. software. We included 197 individuals: 74 index cases and 123 relatives. CLMs were diagnosed in 65% of patients, ULMs in 90% of women, RCys in 37% and RCC in 10.9%. Twenty-seven different pathogenic variants were detected, 12 (44%) of them not reported previously. Patients with missense pathogenic variants showed higher frequencies of CLMs, ULMs and RCys, than those with loss-of-function variants (p = 0.0380, p = 0.0015 and p = 0.024, respectively). This is the first report of patients with HLRCC from Spain. The frequency of RCCs was lower than those reported in the previously published series. Individuals with missense pathogenic variants had higher frequencies of CLMs, ULMs and RCys

Genetic and clinical characterization of a novel FH founder mutation in families with hereditary leiomyomatosis and renal cell cancer syndrome

Hereditary leiomyomatosis and renal cell cancer syndrome is a rare autosomal dominant hereditary syndrome. Previously, we published the largest cohort of FH mutation carriers in Spain and observed a highly recurrent missense heterozygous variant, FH(NM_000143.4):c.1118A > G p.(Asn373Ser), in 104 individuals from 31 apparently unrelated families. Here, we aimed to establish its founder effect and characterize the associated clinical phenotype. Results Haplotype analysis confirmed that families shared a common haplotype (32/38 markers) spanning 0.61-0.82 Mb, indicating this recurrent variant was inherited from a founder ancestor. Cutaneous and uterine leiomyomatosis were diagnosed in 64.6% (64/99) and 98% (50/51) of patients, respectively, and renal cell cancer was present in 10.4% (10/96). The pathogenic FH_c.1118A > G variant is a Spanish founder mutation that originated 12-26 generations ago. We estimate that the variant may have appeared between 1370 and 1720. Individuals carrying this founder mutation had similar frequency of renal cell cancer and a higher frequency of renal cysts and leiomyomas than those in other cohorts of this syndrome. Conclusions In the Spanish province of Alicante there is a high prevalence of HLRCC because of the founder mutation FH c.1118A > G; p.(Asn373Ser). The characterization of founder mutations provides accurate and specific information regarding their penetrance and expressivity. In individuals with suspected HLRCC from the province of Alicante, genetic testing by direct analysis of the founder FH c.1118A > G; p.(Asn373Ser) mutation may be a faster and more efficient diagnostic tool compared with complete gene sequencing