The Structure of a Biologically Active Influenza Virus Ribonucleoprotein Complex

The influenza viruses contain a segmented, single-stranded RNA genome of negative polarity. Each RNA segment is encapsidated by the nucleoprotein and the polymerase complex into ribonucleoprotein particles (RNPs), which are responsible for virus transcription and replication. Despite their importance, information about the structure of these RNPs is scarce. We have determined the three-dimensional structure of a biologically active recombinant RNP by cryo-electron microscopy. The structure shows a nonameric nucleoprotein ring (at 12 Å resolution) with two monomers connected to the polymerase complex (at 18 Å resolution). Docking the atomic structures of the nucleoprotein and polymerase domains, as well as mutational analyses, has allowed us to define the interactions between the functional elements of the RNP and to propose the location of the viral RNA. Our results provide the first model for a functional negative-stranded RNA virus ribonucleoprotein complex. The structure reported here will serve as a framework to generate a quasi-atomic model of the molecular machine responsible for viral RNA synthesis and to test new models for virus RNA replication and transcription

Colgajo de vasto lateral de pedículo distal para cobertura de lesiones complejas de rodilla

El tratamiento de las heridas complejas articulares se muestra siempre complicado, el abordaje razonable conjunto por traumatólogos y cirujanos plásticos mejora el resultado clínico, funcional y estético. Presentamos el caso de un paciente politraumatizado con pérdida de sustancia a nivel de la rodilla secundaria a artritis séptica, necrosis de patela y exposición de cóndilos femorales que fue tratado mediante colgajo muscular de vastus lateralis de pedículo distal. El vastus lateralis permite la cobertura eficiente de l articulación expuesta sin afectar la extensión de la rodilla y dejando mínima secuela estética.The treatment of the joint complex wounds is difficult. A good clinic, functional and aesthetic outcome is possible when multidisciplinary approach by orthopaedics and plastic surgeons is carried out. We report a case of a patient with multiple fractures who presents a severe loss of soft tissue at the knee. This lesion was due to a septic arthritis with patellar necrosis and condillar exposition. The treatment was made by a distally pedicled vastus lateralis muscular flap. This flap lets efficient coverage for the exposed joint as well as the integrity of the extensión knee function without aesthetic seque

Identification and molecular biodiversity of autochthonous grapevine cultivars in the ‘Comarca del Bierzo’, León, Spain

A prospecting work at the ‘Comarca del Bierzo’ in Spain has been carried out in order to evaluate the existing biodiversity and detect the neglected and endangered minor grapevine cultivars of the zone. As a result, a total of 79 different accessions were collected and studied using STMS markers for preliminary variety identification, that was confirmed on the base of ampelographic observations. Several synonymies and homonymies were detected. A total of 33 different cultivars were identified. Comparison of the genotypes, allelic frequencies and allelic sizes are presented. Conservation of the endangered cultivars is highly recommended

Prospection and identification of grapevine varieties cultivated in north Portugal and northwest Spain

A broad prospection of grapevine plants was carried out in northwest Spain and north of Portugal during the period from 2002 to 2009. It included zones located in Galicia as well as in the provinces of León, Zamora and Salamanca in Spain and the region delimited by the right side of the Douro river and the spanish border in Portugal. A total of 669 accessions were sampled, studied and identified, both by using the six microsatellite loci included in the OIV descriptors list, plus ampelographic characters. As a result of the study a total of 53 different grapevine varieties were identified. Most of them correspond to minor varieties that are present in the zone. In many cases the same variety was detected in both countries, although denominated with different names that are synonymous. The molecular markers that were analyzed are those included in the OIV 801 to 806 descriptors, and correspond to the following loci: VVS2, VVMD5, VVMD7, VVMD27, ssrVrZAG62 and ssrVrZAG79. Allele sizes of the studied varieties are recorded. Relationships among these varieties seem to be possible. Recommended priority names for the studied varieties both in Spain and in Portugal are listed, in order to avoid misidentifications and reduce the incidence of homonymies.

Direct specific isotopic analysis of compounds released by pyrolysis (Py-CSIA): Novel applications in paleoenvironmental studies

Poster J.03 presentado en el The 40th International Symposium on Capillary Chromatography and 13th GCxGC Symposium May 29-June 03, (2016) Riva del Garda Fierecongressi, Riva del Garda, Italyisotopic analysis (Py-CSIA) and analytical pyrolysis (Py-GC/MS) opens new windows of information and is particularly useful to study solid materials that are not soluble and therefore not amenable by conventional GC/MS techniques. Py-CSIA is a rather novel hyphenated technique that combines the chromatographic separation of compounds released by pyrolysis (Py-GC) with an isotope ratio mass spectrometer (IRMS). The technique allows the measurement of stable isotope proportions i.e., δ13C, δ15N and δ2H, δ18O in specific compounds released by pyrolysis. The sample preparation is minimized and a molecular fingerprinting of the material is achieved.N

Factors involved in soil organic matter stabilization in Peruvian Amazonian soils (Ucayali region) and the molecular composition of extractable lipids

Poster E1012b presentado en el 27th International Meeting on Organic Geochemistry September 13–18, 2015, Prague, Czech RepublicAmazonian ecosystems are considered important sinks for atmospheric CO2 on Earth. It is therefore desirable to preserve their biodiversity and productivity. In this communication an assessment of the influence of different agro-forestry practices on soil C storage in representative ecosystems from the Peruvian Ucayali region is approached by analyzing the soil lipid fraction. In fact, this organic matter fraction is an important source of analytical surrogates of soil C stabilization and accumulation processes (Naafs at al., 2004; Poulenard et al., 2004; Rumpel et al., 2004). Top (0–20 cm) and subsoil (20–40 cm) samples were taken from the Amazonian Ucayali region (Pucallpa, Perú), ranging from recent alluvial soils in muddy zones of riverside areas (wetlands referred to as ‘mud’, ‘beach’ and low ‘restinga’ soils) to more developed soils located in medium and higher riverine terraces, as well as in hills. The lipid fraction was Soxhlet extracted with a mixture of ichloromethane:methanol (2:1 by vol.), saponified and divided into neutral and acid subfractions. The acid fractions were then sequentially methylated and silylated prior to the chromatographic analysis (González-Vila et al., 2003). Neutral and acid sub-fractions were separated and the major compounds were identified by gas chromatography-mass spectrometry using an HP G1800A GCD System (electron impact detector at 70 eV) equipped with a DB-5 fused silica capillary column (30 m × 0.32 mm i.d., film thickness 0.25 µm) and using He as carrier gas at a flow rate of 1.5 mL min-1. The chromatographic oven temperature was programmed to increase from 40 to 100 ºC at 30 ºC min-1 and then to 300 ºC at 6 ºC min -1. The values of soil organic matter and lipids ranged from 25.8 to 5.8 g kg-1, and from 5.2 to 0.2 g kg-1, respectively. This variability as well as the differences in lipid molecular composition could suggest differences in the soils’ use and management practices (Van Bergen et al.,1996). The main families of signature lipids detected in the soil extracts were n-alkanes, linear isoprenoids, cyclic alkanes, n-fatty acids, branched fatty acids, unsaturated fatty acids, hydroxyacids, n-alcohols, ketones, polycyclic hydrocarbons and sterols. Very significant differences in qualitative and quantitative lipid composition were found in terms of soil depth. This effect was more pronounced in the soils from the riverine high terraces devoted to agro-forestry practices (Figs. 1 and 2). In addition, lipidic compounds of high molecular weight (i.e., di- and triterpenes) were detected as regular components of the free lipid fractions from the different ecosystems (wetlands, terraces and hills) reflecting specific vegetation and soil-use influences. Xenobiotic compounds (naphthenic acids) detected within the unresolved chromatographic ‘hump’ (highly unresolved mixture of cyclic or branched hydrocarbons) were also observed, indicating anthropogenic contamination by mineral oils. Likewise, dialkyl phthalates (Phth) from plasticizers were also detected.Projects CGL2012-38655-C04-01 and CGL2008-04296 and fellowship BES-2013-062573 given to N.T.J.M by the Spanish Ministry of Economy and Competitiveness. Dr. J.M.R. is the recipient of a fellowship from the JAE-Doc subprogram financed by the CSIC and the European Social Fund.Peer Reviewe

Comorbidity and osteoporotic fracture: approach through predictive modeling techniques using the OSTEOMED registry

Purpose To examine the response to anti-osteoporotic treatment, considered as incident fragility fractures after a minimum follow-up of 1 year, according to sex, age, and number of comorbidities of the patients. Methods For this retrospective observational study, data from baseline and follow-up visits on the number of comorbidities, prescribed anti-osteoporotic treatment and vertebral, humerus or hip fractures in 993 patients from the OSTEOMED registry were analyzed using logistic regression and an artificial network model. Results Logistic regression showed that the probability of reducing fractures for each anti-osteoporotic treatment consid- ered was independent of sex, age, and the number of comorbidities, increasing significantly only in males taking vitamin D (OR = 7.918), patients without comorbidities taking vitamin D (OR = 4.197) and patients with ≥ 3 comorbidities taking calcium (OR = 9.412). Logistic regression correctly classified 96% of patients (Hosmer–Lemeshow = 0.492) compared with the artificial neural network model, which correctly classified 95% of patients (AUC = 0.6). Conclusion In general, sex, age and the number of comorbidities did not influence the likelihood that a given anti-osteoporotic treatment improved the risk of incident fragility fractures after 1 year, but this appeared to increase when patients had been treated with risedronate, strontium or teriparatide. The two models used classified patients similarly, but predicted differently in terms of the probability of improvement, with logistic regression being the better fit

Multigene germline testing usefulness instead of BRCA1/2 single screening in triple negative breast cancer cases

Producción CientíficaTriple negative breast cancer is considered as the worst aggressive subtype with poor prognosis. Recent studies suggest a hereditary component is involve in TNBC development, especially in young patients. However, genetic spectrum remains unclear. Our purpose was to evaluate the usefulness of multigene panel testing in triple negative patients respect overall breast cancer cases as well as contributing to elucidate which genes are most implicated in TNBC development with respect to the remaining breast cancer subtypes. A breast cancer patients sample comprised of 100 triple negative breast cancer patients and 100 other breast cancer subtypes patients were analyzed by Next-Generation Sequencing using an On-Demand panel which included 35 predisposition cancer genes associated with inherited cancer susceptibility. Triple negative breast cancer patients obtained a higher percentage of germline variant carriers. ATM, PALB2, BRIP1 and TP53 were the most non-BRCA mutated genes. Moreover, triple negative breast cancer patients without family history related which proved to be carriers were diagnosed at significant earlier age. As conclusion, our study reinforces the usefulness of multigene panel testing in breast cancer cases but specifically in those with triple negative subtype regardless family history.Junta de Castilla y León. Dirección Regional de Salud de Castilla y León (GRS/2180/A/2020 y GRS/2351/A/2021

Plasma MicroRNA Signature Validation for Early Detection of Colorectal Cancer

OBJECTIVES: Specific microRNA (miRNA) signatures in biological fluids can facilitate earlier detection of the tumors being then minimally invasive diagnostic biomarkers. Circulating miRNAs have also emerged as promising diagnostic biomarkers for colorectal cancer (CRC) screening. In this study, we investigated the performance of a specific signature of miRNA in plasma samples to design a robust predictive model that can distinguish healthy individuals from those with CRC or advanced adenomas (AA) diseases. METHODS: Case control study of 297 patients from 8 Spanish centers including 100 healthy individuals, 101 diagnosed with AA, and 96 CRC cases. Quantitative real-time reverse transcription was used to quantify a signature of miRNA (miRNA19a, miRNA19b, miRNA15b, miRNA29a, miRNA335, and miRNA18a) in plasma samples. Binary classifiers (Support Vector Machine [SVM] linear, SVM radial, and SVM polynomial) were built for the best predictive model. RESULTS: Area under receiving operating characteristic curve of 0.92 (95% confidence interval 0.871-0.962) was obtained retrieving a model with a sensitivity of 0.85 and specificity of 0.90, positive predictive value of 0.94, and negative predictive value of 0.76 when advanced neoplasms (CRC and AA) were compared with healthy individuals. CONCLUSIONS: We identified and validated a signature of 6 miRNAs (miRNA19a, miRNA19b, miRNA15b, miRNA29a, miRNA335, and miRNA18a) as predictors that can differentiate significantly patients with CRC and AA from those who are healthy. However, large-scale validation studies in asymptomatic screening participants should be conducted

Clinical features and health-related quality of life in adult patients with mucopolysaccharidosis IVA: the Spanish experience

Background: Mucopolysaccharidosis (MPS) IVA or Morquio A syndrome is a progressive and disabling disease characterized by a deficiency of the enzyme N-acetylgalactosamine-6-sulphate sulphatase. Its clinical presentation is very heterogeneous and poorly understood in adults. The aim of this study was to describe the clinical manifestations of MPS IVA in adult patients in Spain and to assess their health-related quality of life (HRQoL). Results: Thirty-three patients from nine reference centres participated in the study. The median age was 32 (interquartile range [IQR]: 20.5–40.5) years. The phenotype was classical in 54.5% of patients, intermediate in 33.3% of patients, and non-classical in 12.1% of patients. The most common clinical manifestation was bone dysplasia, with a median height of 118 (IQR: 106–136) cm. Other frequent clinical manifestations were hearing loss (75.7%), ligamentous laxity (72.7%), odontoid dysplasia (69.7%), limb deformities that required orthopaedic aids (mainly hip dysplasia and genu valgus) (63.6%), and corneal clouding (60.6%). In addition, 36.0% of patients had obstructive sleep apnoea/hypopnoea syndrome and 33.3% needed non-invasive ventilation. Cervical surgery and varisation osteotomy were the most common surgical interventions (36.4% each). Almost 80% of patients had mobility problems and 36.4% used a wheelchair at all times. Furthermore, 87.9% needed help with self-care, 33.3% were fully dependent, and 78.8% had some degree of pain. HRQoL according to the health assessment questionnaire was 1.43 (IQR: 1.03–2.00) in patients with the non-classical phenotype, but 2.5 (IQR: 1.68–3.00) in those with the classical phenotype. Seven patients were initiated on enzyme replacement therapy (ERT), but two of them were lost to follow-up. Lung function improved in four patients and slightly worsened in one patient. The distance achieved in the six-minute walk test increased in the four patients who could perform it. HRQoL was better in patients treated with elosulfase alfa, with a median (IQR) of 1.75 (1.25–2.34) versus 2.25 (1.62–3.00) in patients not treated with ERT. Conclusions: The study provides real-world data on patients with MPS IVA. Limited mobility, difficulties with self-care, dependence, and pain were common, together with poor HRQoL. The severity and heterogeneity of clinical manifestations require the combined efforts of multidisciplinary teams