Ultra-deep pyrosequencing detects conserved genomic sites and quantifies linkage of drug-resistant amino acid changes in the hepatitis B virus genome

Background: Selection of amino acid substitutions associated with resistance to nucleos(t)ide-analog (NA) therapy in the hepatitis B virus (HBV) reverse transcriptase (RT) and their combination in a single viral genome complicates treatment of chronic HBV infection and may affect the overlapping surface coding region. In this study, the variability of an overlapping polymerase-surface region, critical for NA resistance, is investigated before treatment and under antiviral therapy, with assessment of NA-resistant amino acid changes simultaneously occurring in the same genome (linkage analysis) and their influence on the surface coding region. Methodology/Principal Findings: Serum samples obtained from chronic HBV-infected patients at pre-treatment and during sequential NA treatment with lamivudine, adefovir, and entecavir were analyzed by ultra-deep pyrosequencing (UDPS) using the GS-FLX platform (454 Life Sciences-Roche). The pre-treatment HBV quasispecies was not enriched with NA-resistant substitutions. The frequencies of this type of substitutions at pre-treatment did not predict the frequencies observed during lamivudine treatment. On linkage analysis of the RT region studied, NA-resistant HBV variants (except for rtA181T) were present in combinations of amino acid substitutions that increased in complexity after viral breakthrough to entecavir, at which time the combined variant rtL180M-S202G-M204V-V207I predominated. In the overlapping surface region, NA-resistant substitutions caused selection of stop codons in a significant percentage of sequences both at pre-treatment and during sequential treatment; the rtA181T substitution, related to sW172stop, predominated during treatment with lamivudine and adefovir. A highly conserved RT residue (rtL155), even more conserved than the essential residues in the RT catalytic motif YMDD, was identified in all samples. Conclusions: UDPS methodology enabled quantification of HBV quasispecies variants, even those harboring complex combinations of amino acid changes. The high percentage of potentially defective genomes, especially in the surface region, suggests effective trans-complementation of these variants. © 2012 Rodriguez-Frías et al.Centro para el Desarrollo Tecnológico Industrial; Ministry of Economy and CompetitivenessPeer Reviewe

Which region and which sector leads the circular economy? CEBIX, a multivariant index based on business actions

Consejeria de Educacion, Junta de Castilla y Leon [Grant/Award No. SA069G18] ; Ministerio de Ciencia, Innovacion y Universidades [Grant/Award No. RTI2018-093423-B-I00] ; Universidad de Salamanca [Grant/Award No. USAL2017-DISAQ] ; y Junta de Castilla y Leon y Fondo Europeo de Desarrollo Regional [Grant/Award No. CLU-2019-03 Uni-dad de Excelencia "Gestion Economica para la Sostenibilidad" (GECOS) ] ; and Universidad de Salamanca [Grant/Award Open Access] .The circular economy encompasses a sustainable economic model based on a production, consumption, distribution and maintenance process that reuses as much as possible. In this research, the two-step composite Circular Economy Business Index was created, based on 17 environmental practices that companies have implemented to reduce the generation of waste and emissions and to increase the reuse and efficiency of materials and energy, among other actions. The use of a sample of 26,783 companies from 49 countries and 10 sectors for the period 2014–2019 allowed the aggregation of these initiatives at the country and industry levels. In this sense, our results show less progress in the circular transformation worldwide and can be used in the design of policies aimed at promoting changes in production and consumption systems in specific geographic or industrial contexts.Junta de Castilla y Leon SA069G18Ministerio de Ciencia, Innovacion y Universidades RTI2018-093423-B-I00Universidad de Salamanca y Junta de Castilla y Leon y Fondo Europeo de Desarrollo Regional CLU-2019-03 Uni-dad de Excelencia "Gestion Economica para la Sostenibilida

Linoleic acid-derived oxylipins and isoprostanes plasma levels are influenced by 1,25-Dihydroxyvitamin D levels in middle-aged sedentary adults: The FIT-AGEING study

Introduction: Vitamin D – concretely its active form 1,25-dihydroxyvitamin D (1,25(OH)2D) – maintains several physiological processes. Oxylipins are oxidized lipids derived from ω-6 and ω-3 polyunsaturated fatty acids involved in inflammation. Little is known about the association of 1,25(OH)2D with inflammatory parameters in middle-aged populations – who could be at risk of vitamin D deficiency –. The aim of this study was to investigate the relationship between 1,25(OH)2D plasma levels with circulating white blood cells, platelets counts and oxylipins levels. Materials and methods: A total of 74 (53 % women) middle-aged (40–65 years old) adults were recruited for this cross-sectional study. 1,25(OH)2D plasma levels were measured using an immunochemiluminometric assay. White blood cells and platelets were analyzed by hemocytometry. ω-6 and ω-3 oxylipins plasma levels were measured using liquid chromatography - tandem mass spectrometry. Simple and multiple linear regression models, and Pearson correlation analyses, were performed to study the association of 1,25(OH)2D levels with WBC and platelets counts, and oxylipins, respectively. Results: 1,25(OH)2D plasma levels were positively related with linoleic acid-derived oxylipins and isoprostanes plasma levels, whereas an inverse relationship with dihomo-γ-linolenic acid/linoleic acid and arachidonic acid/ linoleic acid ratios was unveiled. No significant associations were observed for circulating ω-3 oxylipins, white blood cells levels or platelets count. Conclusions: Linoleic acid-derived oxylipins and isoprostanes plasma levels may be influenced by 1,25(OH)2D plasma levels. Further investigations are needed to elucidate the impact of other vitamin D forms upon circulating oxylipins levels.Spanish GovernmentChina Scholarship Council 201607060017 20170706001

El rol de la consistencia del mensaje y de la estrategia de marca en el capital de marca de un destino

La estrategia de comunicación seguida para difundir un destino turístico contribuye a su competitividad. El objetivo de esta investigación es conocer la importancia de la consistencia de los mensajes entre medios y de la estrategia de posicionamiento de marca en la difusión de un destino turístico. Con esta finalidad, se ha diseñado un experimento manipulando la consistencia del mensaje (alta consistencia vs baja consistencia de mensajes) y la estrategia de posicionamiento seguida por el destino (marca destino-único vs marca destino-múltiple). Los resultados obtenidos indican que la generación de una comunicación con mensajes altamente consistentes así como un posicionamiento basada en una estrategia de marca-única contribuyen a aumentar el Customer Based Destination Brand Equity. Además, se comprueba que el efecto de posicionamiento puede estar moderado por la consistencia de los mensajes a los que está expuesto el consumidor. Los resultados alcanzados suponen un avance en la literatura académica y una aportación para el sector profesional.A well-implemented communication strategy to raise the profile of a tourist destination contributes to its competitiveness. The aim of the present study is to understand the importance of message consistency across different media, and of the brand positioning strategy, in the promotion of a tourist destination. An experiment is conducted, manipulating (a) the degree of message consistency (high vs. low) and (b) the positioning strategy followed by the destination (sole-destination brand vs. multiple-destination brand). The results of the investigation suggest that communications based on highly consistent messages, and a sole-destination brand positioning strategy, contribute to increasing Customer-Based Destination Brand Equity. Further, the work demonstrates that the effect of positioning may be moderated by the degree of consistency between the messages to which the consumer is exposed. The findings represent a contribution to both the academic literature and to the professional sector.Ministerio de Economía y Competitividad ECO-2012-39217Junta de Andalucía P11 SEJ-810

Patología implanto-endodóncica: concepto, tipos, diagnóstico, tratamiento y prevención.

La patología implanto-endodóncica (PIE) está descrita en la literatura implantológica como una de las causas de periimplantitis apical, entendida como la lesión osteolítica en la región apical del implante, con normal osteointegración de su porción coronal, provocada por la infección por contigüidad a partir de la lesión periapical del diente adyacente. Pero el concepto de PIE no sólo abarca la periimplantitis retrógrada por contaminación diente-a-implante, sino también los procesos inflamatorios periapicales en dientes adyacentes al implante por contaminación implante-adiente, cuando la colocación del implante provoca la necrosis del diente adyacente y la consiguiente periodontitis apical. Incluso podríamos incluir dentro de la PIE los casos de periimplantitis apical en implantes postextracción provocada por la infección residual presente en el alvéolo de un diente extraído con periodontitis apical. En definitiva, la PIE incluye las lesiones endodóncicas e implantarias apicales que son el resultado de infecciones residuales o por contigüidad entre diente e implante. En esta revisión bibliográfica se define y clasifica la PIE, repasándose la casuística publicada así como su influencia en el resultado del tratamiento implantológico

Association between circulating alpha-1 antitrypsin polymers and lung and liver disease

Alpha-1 antitrypsin deficiency; Circulating polymers; EmphysemaDeficiència d'alfa-1 antitripsina; Polímers circulants; EmfisemaDeficiencia de alfa-1 antitripsina; Polímeros circulantes; EnfisemaBackground Alpha-1 antitrypsin deficiency (AATD) is considered one of the most common genetic diseases and is characterised by the misfolding and polymerisation of the alpha-1 antitrypsin (AAT) protein within hepatocytes. The relevance of circulating polymers (CP) of AAT in the pathogenesis of lung and liver disease is not completely understood. Therefore, the main objective of our study was to determine whether there is an association between the levels of CP of AAT and the severity of lung and liver disease. Method This was a cross-sectional study in patients with different phenotypes of AATD and controls. To quantify CP, a sandwich ELISA was performed using the 2C1 monoclonal antibody against AAT polymers. Sociodemographic data, clinical characteristics, and liver and lung parameters were collected. Results A cohort of 70 patients was recruited: 32 Pi*ZZ (11 on augmentation therapy); 29 Z-heterozygous; 9 with other genotypes. CP were compared with a control group of 47 individuals (35 Pi*MM and 12 Pi*MS). ZZ patients had the highest concentrations of CP (p < 0.001) followed by Z heterozygous. The control group and patients with Pi*SS and Pi*SI had the lowest CP concentrations. Pi*ZZ also had higher levels of liver stiffness measurements (LSM) than the remaining AATD patients. Among patients with one or two Z alleles, two patients with lung and liver impairment showed the highest concentrations of CP (47.5 µg/mL), followed by those with only liver abnormality (n = 6, CP = 34 µg/mL), only lung (n = 18, CP = 26.5 µg/mL) and no abnormalities (n = 23, CP = 14.3 µg/mL). Differences were highly significant (p = 0.004). Conclusions Non-augmented Pi*ZZ and Z-patients with impaired lung function and increased liver stiffness presented higher levels of CP than other clinical phenotypes. Therefore, CP may help to identify patients more at risk of developing lung and liver disease and may provide some insight into the mechanisms of disease.This study was supported by funding from Grifols to the Catalan Center for Research in Alpha-1 antitrypsin deficiency of the Vall d’Hebron Research Institute (VHIR) in the Vall d’Hebron Barcelona Hospital Campus, Barcelona, Spain and with a grant from the Fundació Catalana de Pneumologia (FUCAP)

New variants of alpha-1-antitrypsin : structural simulations and clinical expression

Alpha-1 antitrypsin deficiency (AATD) is characterized by reduced serum levels of the AAT protein and predisposes to liver and lung disease. The characterization at structural level of novel pathogenic SERPINA1 mutants coding for circulating AAT could provide novel insights into the mechanisms of AAT misfolding. The present study aimed to provide a practical framework for the identification and analysis of new AAT mutations, combining structural simulations and clinical data. We analysed a total of five mutations (four not previously described) in a total of six subjects presenting moderate to severe AATD: Gly95Alafs*18, Val210Glu, Asn247Ser, Pi*S + Asp341His and Pi*S + Leu383Phe + Lys394Ile. Clinical data, genotyping and phenotyping assays, structural mapping, and conformational characterization through molecular dynamic (MD) simulations were developed and combined. Newly discovered AAT missense variants were localized both on the interaction surface and the hydrophobic core of the protein. Distribution of mutations across the structure revealed Val210Glu at the solvent exposed s4C strand and close to the "Gate" region. Asn247Ser was located on the accessible surface, which is important for glycan attachment. On the other hand, Asp341His, Leu383Phe were mapped close to the "breach" and "shutter" regions. MD analysis revealed the reshaping of local interactions around the investigated substitutions that have varying effects on AAT conformational flexibility, hydrophobic packing, and electronic surface properties. The most severe structural changes were observed in the double- and triple-mutant (Pi*S + Asp341His and Pi*S + Leu383Phe + Lys394Ile) molecular models. The two carriers presented impaired lung function. The results characterize five variants, four of them previously unknown, of the SERPINA1 gene, which define new alleles contributing to the deficiency of AAT. Rare variants might be more frequent than expected, and therefore, in discordant cases, standardized screening of the S and Z alleles needs complementation with gene sequencing and structural approaches. The utility of computational modelling for providing supporting evidence of the pathogenicity of rare single nucleotide variations is discussed. The online version contains supplementary material available at 10.1186/s12931-022-02271-8

The intake of fried virgin olive or sunflower oils differentially induces oxidative stress in rat liver microsomes

Published online by Cambridge University Press: 09 March 2007This work was supported by CICYT project ALI91-1113- C03-01. M. Battino was a visiting scientist at Granada University, thanks to Ministerio de Educación y Ciencia, Madrid. J.L. Quiles and M.C. Ramirez-Tortosa received a short-term fellowship by Ancona University.The effects of non-fried and fried virgin olive and sunflower oils on rat liver microsomal compositional features have been investigated. In addition, plasma antioxidants (α-tocopherol and ubiquinone 9) were investigated as well as the possible oxidative modifications suffered by virgin olive and sunflower oils during the frying process. The frying process decreased the content of α-tocopherol and phenolics in the oils and increased total polar materials. Sunflower oil was affected to a greater extent than olive oil. In rats, the intake of fried oil led to higher levels of lipid peroxidation and a lower concentration of plasma antioxidants. Microsomal fatty acid and antioxidant profiles were also altered. It seems that a strong relationship exists between the loss of antioxidants and the production of toxic compounds in the oils after frying and the extent of the peroxidative events in microsomes, which were also different depending on the fat source. The highly unsaturated sunflower oil was less resistant to the oxidative stress produced by frying and led to a higher degree of lipid peroxidation in liver microsomes in vivo than virgin olive oil.CICYT project ALI91-1113- C03-01.

A cohort of patients with COVID-19 in a major teaching hospital in Europe

Artículo escrito por un elevado número de autores, solo se referencian el que aparece en primer lugar, el nombre del grupo de colaboración, si le hubiere, y los autores pertenecientes a la UAMArtículo escrito en nombre del COVID@HULP Working GroupSince the confirmation of the first patient infected with SARS-CoV-2 in Spain in January 2020, the epidemic has grown rapidly, with the greatest impact on the region of Madrid. This article describes the first 2226 adult patients with COVID-19, consecutively admitted to La Paz University Hospital in Madrid. Methods: Our cohort included all patients consecutively hospitalized who had a final outcome (death or discharge) in a 1286-bed hospital of Madrid (Spain) from 25 February (first case admitted) to 19 April 2020. The data were manually entered into an electronic case report form, which was monitored prior to the analysis. Results: We consecutively included 2226 adult patients admitted to the hospital who either died (460) or were discharged (1766). The patients’ median age was 61 years, and 51.8% were women. The most common comorbidity was arterial hypertension (41.3%), and the most common symptom on admission was fever (71.2%). The median time from disease onset to hospital admission was 6 days. The overall mortality was 20.7% and was higher in men (26.6% vs. 15.1%). Seventy-five patients with a final outcome were transferred to the intensive care unit (ICU) (3.4%). Most patients admitted to the ICU were men, and the median age was 64 years. Baseline laboratory values on admission were consistent with an impaired immune-inflammatory profile. Conclusions: We provide a description of the first large cohort of hospitalized patients with COVID-19 in Europe. Advanced age, male sex, the presence of comorbidities and abnormal laboratory values were more common among the patients with fatal outcome

Cost-effectiveness analysis of an active search to retrieve HCV patients lost to follow-up (RELINK-C strategy) and the impact of COVID-19

Acord transformatiu CRUE-CSICAltres ajuts: Gilead Science