Composition influence on pulmonary delivery of rifampicin liposomes

The effects of lipid concentration and composition on the physicochemical properties, aerosol performance and in vitro toxicity activity of several rifampicin-loaded liposomes were investigated. To this purpose, six liposome formulations containing different amounts of soy phosphatidylcholine and hydrogenated soy phosphatidylcholine, with and without cholesterol and oleic acid, were prepared and fully characterized. Uni- or oligo-lamellar, small (~100 nm), negatively charged (~60 mV) vesicles were obtained. Lipid composition affected aerosol delivery features of liposomal rifampicin; in particular, the highest phospholipid concentration led to a better packing of the vesicular bilayers with a consequent higher nebulization stability. The retention of drug in nebulized vesicles (NER%) was higher for oleic acid containing vesicles (55% ± 1.4%) than for the other samples (~47%). A549 cells were used to evaluate intracellular drug uptake and in vitro toxicity activity of rifampicin-loaded liposomes in comparison with the free drug. Cell toxicity was more evident when oleic acid containing liposomes were used

6 projectes d’espai públic urbà premiats a la ciutat de Barcelona entre els anys 90 i l’actualitat: la mirada acadèmica, la popular i la pròpia

Barcelona és i ha estat una ciutat pionera i exemplar pel que fa al seu urbanisme i espai públic en clau sociològica, territorial i demogràfica, sent fins i tot un model a seguir en altres ciutats europees amb el conegut "Model Barcelona". Ja des de principis de segle, es plantejaven qüestions urbanístiques innovadores que es van veure paralitzades per la guerra civil i la dictadura, però amb les primeres eleccions democràtiques, amb Oriol Bohigas al capdavant com a delegat d'urbanisme, va ressorgir una Barcelona ocupada per les qüestions socials i que posava el focus en un espai públic de qualitat. Es van crear més de cent places i nombrosos parcs en un temps rècord i es va posar especial atenció a l'espai públic d'una Barcelona pluricèntrica i que havia vist augmentada considerablement la seva població a causa de les onades migratòries procedents de la resta de l'Estat. Oriol Bohigas va dirigir l'inici d'un seguit de mesures socials i urbanístiques per dotar a una Barcelona d'escala metropolitana d'un espai públic identitari i pròsper. Posteriorment, la concessió dels Jocs Olímpics del 92 van suposar la culminació d'importants projectes d'infraestructures i la creació de més places, parcs i equipaments. I més endavant amb la celebració del Fòrum internacional de les cultures, l'any 2004, es planteja un altre ambiciós projecte de transformació urbana i metropolitana que uneixi el districte de Sant Martí amb la ciutat de Sant Adrià del Besòs, buscant activament un urbanisme sostenible. L'any 1999 sorgeix arran d'una exposició del CCCB, el Premi Europeu de l'Espai Públic Urbà on cada dos anys un jurat d'experts internacional premien els projectes d'espai públic urbà de les ciutats europees que s'hi presenten. Així doncs, aquest treball se centra en l'anàlisi de sis projectes premiats a la ciutat de Barcelona durant les tres últimes dècades, fent una comparativa entre dos projectes de cada dècada, buscant els criteris de cada època i analitzant el context històric enBarcelona es y ha sido una ciudad pionera y ejemplar en cuanto a su urbanismo y espacio público en clave sociológica, territorial y demográfica, siendo incluso un modelo a seguir en otras ciudades europeas con el conocido "Model Barcelona". Ya desde principios de siglo, se planteaban cuestiones urbanísticas innovadoras que se vieron paralizadas por la guerra civil y la dictadura, pero con las primeras elecciones democráticas, con Oriol Bohigas al frente como delegado de urbanismo, resurgió una Barcelona ocupada por las cuestiones sociales y que ponía el foco en un espacio público de calidad. Se crearon más de cien plazas y numerosos parques en un tiempo récord y se puso especial atención al espacio público de una Barcelona pluricéntrica y que había visto aumentada considerablemente su población debido a las olas migratorias procedentes del resto de el Estado. Oriol Bohigas dirigió el inicio de una serie de medidas sociales y urbanísticas para dotar a una Barcelona de una escala metropolitana de un espacio público identitario y próspero. Posteriormente, la concesión de los Juegos Olímpicos del 92 supuso la culminación de importantes proyectos de infraestructuras y la creación de más plazas, parques y equipamientos. Y más adelante con la celebración del Foro internacional de las culturas, en 2004, se plantea otro ambicioso proyecto de transformación urbana y metropolitana que una el distrito de Sant Martí con la ciudad de Sant Adrià del Besòs, buscando activamente un urbanismo sostenible . En 1999 surge a raíz de una exposición del CCCB, el Premio Europeo del Espacio Público Urbano donde cada dos años un jurado de expertos internacional premian los proyectos de espacio público urbano de las ciudades europeas que se presentan. Así pues, este trabajo se centra en el análisis de seis proyectos premiados en la ciudad de Barcelona durante las tres últimas décadas, haciendo una comparativa entre dos proyectos de cada década, buscando los criterios de cada época y analizandoBarcelona is and has been a pioneering and exemplary city in terms of its urban planning and public space in a sociological, territorial and demographic level. It’s been a model for other European cities to pursue, even so with the wellknown "Barcelona Model". Since the beginning of the century, innovative urban planning questions were raised but were paralyzed by the civil war and the dictatorship. Through the first democratic elections, with Oriol Bohigas at the head as urban planning delegate, Barcelona remerged as a city engaged with social issues focused on the quality of public space. More than a hundred squares and numerous parks were created in record time. Special attention was paid to the public space of a pluricentric Barcelona which had seen its population increase considerably due to waves of migration from the rest of the State. Oriol Bohigas led the start of a series of social and urban planning measures to provide a metropolitan-scale Barcelona with an identarian and prosperous public space. Subsequently, the concession of the 1992 Olympic Games led to the completion of important infrastructure projects and the creation of more squares, parks and equipment. Later, with the celebration of the Universal Forum of Cultures, in 2004, another ambitious project of urban and metropolitan transformation was proposed that unites the district of Sant Martí with the city of Sant Adrià del Besòs, actively seeking a sustainable urbanism. In 1999, the European Award for Urban Public Space emerged as a result of an exhibition by the CCCB. Every two years an international jury of experts awards some of the urban public space projects of the European cities that are presented. Therefore, this work focuses on the analysis of six awarded projects in the city of Barcelona during the last three decades, making a comparison between two projects from each decade, looking for the criteria of each era and analysing the historical context of its construction. The aim, among o

Predicted impact of the viral mutational landscape on the cytotoxic response against SARS-CoV-2

The massive assessment of immune evasion due to viral mutations that increase COVID-19 susceptibility can be computationally facilitated. The adaptive cytotoxic T response is critical during primary infection and the generation of long-term protection. Here, potential HLA class I epitopes in the SARS-CoV-2 proteome were predicted for 2,915 human alleles of 71 families using the netMHCIpan EL algorithm. Allele families showed extreme epitopic differences, underscoring genetic variability of protective capacity between humans. Up to 1,222 epitopes were associated with any of the twelve supertypes, that is, allele clusters covering 90% population. Next, from all mutations identified in ~118,000 viral NCBI isolates, those causing significant epitope score reduction were considered epitope escape mutations. These mutations mainly involved non-conservative substitutions at the second and C-terminal position of the ligand core, or total ligand removal by large recurrent deletions. Escape mutations affected 47% of supertype epitopes, which in 21% of cases concerned isolates from two or more sub-continental areas. Some of these changes were coupled, but never surpassed 15% of evaded epitopes for the same supertype in the same isolate, except for B27. In contrast to most supertypes, eight allele families mostly contained alleles with few SARS-CoV-2 ligands. Isolates harboring cytotoxic escape mutations for these families co-existed geographically within sub-Saharan and Asian populations enriched in these alleles according to the Allele Frequency Net Database. Collectively, our findings indicate that escape mutation events have already occurred for half of HLA class I supertype epitopes. However, it is presently unlikely that, overall, it poses a threat to the global population. In contrast, single and double mutations for susceptible alleles may be associated with viral selective pressure and alarming local outbreaks. The integration of genomic, geographical and immunoinformatic information eases the surveillance of variants potentially affecting the global population, as well as minority subpopulations.This research was supported by Acción Estratégica en Salud from the ISCIII (https://www.isciii.es), grants MPY 380/18 (to MJM), 388/18 (to DL) and 509/19 (to AJM-G). AJM-G is the recipient of a Miguel Servet contract by the ISCIII. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.S

Interactions between local and global drivers determine long-term trends in boreal forest understorey vegetation

Aim Global change effects on forest ecosystems are increasingly claimed to be context dependent, indicated by interactions between global and local environmental drivers. Most examples of such context dependencies originate from temperate systems, while limited research comes from the boreal biome. Here we set out to test if interactions between climate warming, nitrogen deposition, land-use change resulting in increasing forest density, and soil pH drive long-term changes in understorey vegetation in boreal forests. Location Sweden. Time period 1953-2012. Major taxa studied Vascular plants. Methods We used long-term (50 years) National Forest Inventory data on forest understorey vegetation in Sweden to model the combined effects of climate warming, nitrogen deposition, increase in forest density (tree basal area), and soil pH. Results Our results identify increasing temperature, nitrogen deposition and denser, shadier forest conditions as the main drivers of understorey vegetation changes during this time period. More importantly, we found that these effects varied with local conditions, that is, that the change towards a more nitrophilic understorey vegetation was more pronounced at low than high soil pH. Forest density was an important modulator of nitrogen deposition and temperature increase, with effects generally decreasing with density. Decreased cover of ericaceous dwarf shrubs was driven by both forest density and nitrogen deposition, with a stronger effect at low than at high pH. Main conclusions Our results highlight that to understand forest ecosystems' response to global change, and to make adequate management decisions to mitigate the effects of global change, we need to understand how changes in local environmental factors (forest density and soil pH) interact with global-scale drivers (nitrogen deposition and climate warming). Neglecting such interactions will lead to incorrect estimations of effects. In our case, we would for example, have underestimated the eutrophication effects on acid soils, which constitute a considerable part of the boreal biome

Immunometabolism is a key factor for the persistent spontaneous elite control of HIV-1 infection

Approximately 25% of elite controllers (ECs) lose their virological control by mechanisms that are only partially known. Recently, immunovirological and proteomic factors have been associated to the loss of spontaneous control. Our aim was to perform a metabolomic approach to identify the underlying mechanistic pathways and potential biomarkers associated with this loss of control. Methods: Plasma samples from EC who spontaneously lost virological control (Transient Controllers, TC, n=8), at two and one year before the loss of control, were comparedwith a control group of ECwho persistently maintained virological control during the same follow-up period (Persistent Controllers, PC, n=8). The determination of metabolites and plasma lipids was performed by GC-qTOF and LC-qTOF using targeted and untargeted approaches. Metabolite levels were associated with the polyfunctionality of HIV-specific CD8+T-cell response. Findings: Our data suggest that, before the loss of control, TCs showed a specific circulating metabolomic profile characterized by aerobic glycolytic metabolism, deregulated mitochondrial function, oxidative stress and increased immunological activation. In addition, CD8+ T-cell polyfunctionality was strongly associated with metabolite levels. Finally, valine was the main differentiating factor between TCs and PCs. Interpretation: All these metabolomic differences should be considered not only as potential biomarkers but also as therapeutic targets in HIV infection.Instituto Carlos III PI10/02635 PI13/00796 PI16/00503 PI12/02283 PI16/00684 CPII014/00025 FI14/00431 FI17/00186 INT11/240 INT12/282 INT15/226Fondos Europeos para el Desarrollo Regional PI10/02635 PI13/00796 PI16/00503 PI12/02283 PI16/00684 CPII014/00025 FI14/00431 FI17/00186FEDER PI10/02635 PI13/00796 PI16/00503 PI12/02283 PI16/00684 CPII014/00025 FI14/00431 FI17/00186FEDER PI10/02635 PI13/00796 PI16/00503 PI12/02283 PI16/00684 CPII014/00025 FI14/00431 FI17/00186Programa de Suport als Grups de Recerca 2017SGR948 2014SGR250Gilead Fellowship Program GLD14/293 GLD17/00299Red de Investigación en Sida RD12/0017/0005 RD16/0025/0006 RD12/0017/0029 RD16/0025/0020Junta de Andalucía C-0032/17Generalitat de Catalunya PERIS SLT002/16/0010

Role of Splicing Regulatory Elements and In Silico Tools Usage in the Identification of Deep Intronic Splicing Variants in Hereditary Breast/Ovarian Cancer Genes

Cancer hereditario de mama y ovario; Pseudoexones; Variantes intrónicas profundas spliceogénicasCàncer hereditari de mama i d'ovari; Pseudoexons; Variants intròniques profundes spliceogèniquesHereditary breast ovarian cancer; Pseudoexons; Spliceogenic deep intronic variantsThe contribution of deep intronic splice-altering variants to hereditary breast and ovarian cancer (HBOC) is unknown. Current computational in silico tools to predict spliceogenic variants leading to pseudoexons have limited efficiency. We assessed the performance of the SpliceAI tool combined with ESRseq scores to identify spliceogenic deep intronic variants by affecting cryptic sites or splicing regulatory elements (SREs) using literature and experimental datasets. Our results with 233 published deep intronic variants showed that SpliceAI, with a 0.05 threshold, predicts spliceogenic deep intronic variants affecting cryptic splice sites, but is less effective in detecting those affecting SREs. Next, we characterized the SRE profiles using ESRseq, showing that pseudoexons are significantly enriched in SRE-enhancers compared to adjacent intronic regions. Although the combination of SpliceAI with ESRseq scores (considering ∆ESRseq and SRE landscape) showed higher sensitivity, the global performance did not improve because of the higher number of false positives. The combination of both tools was tested in a tumor RNA dataset with 207 intronic variants disrupting splicing, showing a sensitivity of 86%. Following the pipeline, five spliceogenic deep intronic variants were experimentally identified from 33 variants in HBOC genes. Overall, our results provide a framework to detect deep intronic variants disrupting splicing.This research was funded by the Spanish Instituto de Salud Carlos III (ISCIII) funding an initiative of the Spanish Ministry of Economy and Innovation, partially supported by European Regional Development FEDER Funds, grant numbers PI16/01218 and PI19/01303. AM-F contract is supported by the award ERAPERMED2019-215 granted by AECC FC and by ISCIII thorough AES 2019, both within the ERAPerMed framework”. J.D.-V. contract is supported by the Secretariat for Universities and Research of the Ministry of Business and Knowledge of the Government of Catalonia and the European Social Fund

Multi-Stream Extension of Variational Bayesian HMM Clustering (MS-VBx) for Combined End-to-End and Vector Clustering-based Diarization

Combining end-to-end neural speaker diarization (EEND) with vector clustering (VC), known as EEND-VC, has gained interest for leveraging the strengths of both methods. EEND-VC estimates activities and speaker embeddings for all speakers within an audio chunk and uses VC to associate these activities with speaker identities across different chunks. EEND-VC generates thus multiple streams of embeddings, one for each speaker in a chunk. We can cluster these embeddings using constrained agglomerative hierarchical clustering (cAHC), ensuring embeddings from the same chunk belong to different clusters. This paper introduces an alternative clustering approach, a multi-stream extension of the successful Bayesian HMM clustering of x-vectors (VBx), called MS-VBx. Experiments on three datasets demonstrate that MS-VBx outperforms cAHC in diarization and speaker counting performance.Comment: Accepted at Interspeech 202

Proline cis-​trans isomerization and its implications for the dimerization of analogues of cyclopeptide stylostatin 1: a combined computational and experimental study

Cis and trans proline conformers are often associated with dramatic changes in the biological function of peptides. A slow equilibrium between cis and trans Ile-Pro amide bond conformers occurs in constrained derivatives of the native marine cyclic heptapeptide stylostatin 1 (cyclo-(NSLAIPF)), a potential anticancer agent. In this work, four cyclopeptides, cyclo-(NSTAIPF), cyclo-(KSTAIPF), cyclo-(RSTAIPF) and cyclo-(DSTAIPF), which are structurally related to stylostatin 1, are experimentally and computationally examined in order to assess the effect of residue mutations on the cis-trans conformational ratio and the apparent capacity to form dimeric aggregates. Primarily, cyclo-(KSTAIPF) and cyclo-(RSTAIPF) showed specific trends in circular dichroism, MALDI-TOF and HPLC purification experiments, which suggests the occurrence of peptide dimerization. Meanwhile, the NMR spectrum of cyclo-(KSTAIPF) indicates that this cyclopeptide exists in the two slow-exchange families of conformations mentioned above. Molecular dynamics simulations combined with quantum mechanical calculations have shed light on the factors governing the cis/trans conformational ratio. In particular, we have found that residue mutations affect the internal hydrogen bond pattern which ultimately tunes the cis/trans conformational ratio and that only trans conformers are capable of aggregating due to the shape complementarity of the two subunits