Dexibuprofen biodegradable nanoparticles: one step closer towards a better ocular interaction study

Ocular inflammation is one of the most prevalent diseases in ophthalmology, which can affect various parts of the eye or the surrounding tissues. Non-steroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen, are commonly used to treat ocular inflammation in the form of eye-drops. However, their bioavailability in ocular tissues is very low (less than 5%). Therefore, drug delivery systems such as biodegradable polymeric PLGA nanoparticles constitute a suitable alternative to topical eye administration, as they can improve ocular bioavailability and simultaneously reduce drug induced side effects. Moreover, their prolonged drug release can enhance patient treatment adherence as they require fewer administrations. Therefore, several formulations of PLGA based nanoparticles encapsulating dexibuprofen (active enantiomer of Ibuprofen) were prepared using the solvent displacement method employing different surfactants. The formulations have been characterized and their interactions with a customized lipid corneal membrane model were studied. Ex vivo permeation through ocular tissues and in vivo anti-inflammatory efficacy have also been studied.This work was supported by grant RTI2018-094120-B-I00 from the Spanish Ministry of Economy, Industry and Competitiveness (MINECO) and the European Regional Development Fund. ESL wants to acknowledge the Institute of Nanoscience and Nanotechnology (ART2018 project).info:eu-repo/semantics/publishedVersio

Systematic Collaborative Reanalysis of Genomic Data Improves Diagnostic Yield in Neurologic Rare Diseases

Altres ajuts: Generalitat de Catalunya, Departament de Salut; Generalitat de Catalunya, Departament d'Empresa i Coneixement i CERCA Program; Ministerio de Ciencia e Innovación; Instituto Nacional de Bioinformática; ELIXIR Implementation Studies (CNAG-CRG); Centro de Investigaciones Biomédicas en Red de Enfermedades Raras; Centro de Excelencia Severo Ochoa; European Regional Development Fund (FEDER).Many patients experiencing a rare disease remain undiagnosed even after genomic testing. Reanalysis of existing genomic data has shown to increase diagnostic yield, although there are few systematic and comprehensive reanalysis efforts that enable collaborative interpretation and future reinterpretation. The Undiagnosed Rare Disease Program of Catalonia project collated previously inconclusive good quality genomic data (panels, exomes, and genomes) and standardized phenotypic profiles from 323 families (543 individuals) with a neurologic rare disease. The data were reanalyzed systematically to identify relatedness, runs of homozygosity, consanguinity, single-nucleotide variants, insertions and deletions, and copy number variants. Data were shared and collaboratively interpreted within the consortium through a customized Genome-Phenome Analysis Platform, which also enables future data reinterpretation. Reanalysis of existing genomic data provided a diagnosis for 20.7% of the patients, including 1.8% diagnosed after the generation of additional genomic data to identify a second pathogenic heterozygous variant. Diagnostic rate was significantly higher for family-based exome/genome reanalysis compared with singleton panels. Most new diagnoses were attributable to recent gene-disease associations (50.8%), additional or improved bioinformatic analysis (19.7%), and standardized phenotyping data integrated within the Undiagnosed Rare Disease Program of Catalonia Genome-Phenome Analysis Platform functionalities (18%)

Implementació i desenvolupament d'estudis funcionals per demostrar la patogenicitat de variants de significat incert i identificar nous gens causants de malaltia

[cat] Les malalties metabòliques hereditàries (MMH) són un grup de patologies amb una gran heterogeneïtat clínica, bioquímica i genètica. Actualment se n’han descrit més de 1 800. En l’última dècada la implementació de les tècniques de seqüenciació massiva (seqüenciació de l’exoma complet, WES i seqüenciació del genoma complet, WGS) ha permès identificar la causa genètica de la malaltia en un gran nombre de pacients i descobrir un nombre important de nous gens associats a patologia. Malgrat això, aproximadament la meitat dels pacients roman sense rebre un diagnòstic genètic definitiu. Això es deu, en part, a limitacions tècniques pel que fa a la detecció i priorització de variants candidates, així com a l gran nombre de variants de significat incert (VUS) que s’identifiquen. Per aquests motius cal desenvolupar aproximacions addicionals que ajudin a prioritzar i interpretar l’impacte funcional de les variants identificades per poder-ne determinar la seva patogenicitat. En el context d’aquesta tesi doctoral, hem aplicat una estratègia multiòmica, basada en la combinació de WES/WGS amb la seqüenciació de l’RNA (RNA seq)seq), en una cohort de 65 pacients amb sospita de MMH. L’estratègia utilitzada ha permès orientar el diagnòstic i identificar gens causants de malaltia en un gran nombre de pacients. No obstant això, en molts casos s’ha requerit la realització d’estudis funcionals , la contribució dels quals ha estat essencial per demostrar l’impacte de les variants prioritzades i assolir un diagnòstic genètic definitiu. En concret, la tesi doctoral inclou cinc publicacions originals , quatre de les quals es centr en la caracterització funcional de noves variants genètiques en quatre gens implicats en funcions molt diverses : CCDC186 , PEX1 , PTCD3 i TFRC . En el primer treball hem identificat tres pacients amb variants en CCDC186 i confirmem que aquestes s’associen a un trastorn que presenta una característica combinació d’alteracions endocrines i del sistema nerviós central. En el segon article hem aplicat la combinació de WES, RNA seq i estudis funcionals per acabar amb l’odissea diagnòstica de més 30 anys en una pacient amb deficiència de PEX1 . En el tercer treball hem descrit tres pacients amb variants en PTCD3 , que codifica per una proteïna associada al ribosoma mitocondrial , la disfunció de la qual provoca un trastorn del metabolisme energètic mitocondrial. A més, hem consolidat que les variants en aquest gen s’associen amb l a síndrome de Leigh. En el quart treball hem descrit per primer cop que diferents variants en TFRC poden associar-se a diverses presentacions clíniques: immunodeficiència combinada, anèmia microcítica o cardiomiopatia hipertròfica. Estudis funcionals realitzats en models cel·lulars modificats genèticament mitjançant CRISPR/Cas9 han ampliat les bases fisiopatològiques de la deficiència de TFRC , tot mostrant que, en diferent mesura, les variants identificades en els pacients afecten la viabilitat cel·lular, l a regulació de l’autofàgia i l’homeòstasi del ferro i provoquen una alteració generalitzada de la funció mitocondrial. La cinquena publicació ha consistit en el desenvolupament d’un flux de treball basat en el sistema d’edició genètica CRISPR/Cas9 en cèl·lules HAP1 . L’estratègia proposada permet introduir les variants genètiques identificades en pacients i generar models cel·lulars models cel·lulars knockin per tal d’investigar les seves conseqüències funcionals. . En conclusió, aquesta tesi doctoral emfatitza la importància d’implementar els estudis de validació funcional, així com altres aproximacions òmiques més enllà del WES/WGS al flux diagnòstic de les malalties minoritàries. En aquest sentit, la incorporació de l’RNAl’RNA-seq ha contribuït de forma important al diagnòstic gràcies a la identificació de variants causants de patologia no prioritzades pel WES, així com generant evidències funcionals de l’impacte d’aquelles variants que afecten generant el processament del mRNA.. A més, els estudis funcionals realitzats en material biològic dels pacients o en models cel·lulars modificats genèticament han contribuït en avaluar l’impacte de les variants identificades, tot ajudant a dirigir el diagnòstic i en alguns casos a la presa de decisions decisions clíniques

Obtenció de nanopartícules polimèriques per a l'administració ocular de fàrmacs i estudis d'interacció amb tècniques biofísiques

Ageing is a biological process that leads to a decrease in physiological functions, and is the main risk factor for most common diseases in developed countries. The process causes modifications in biological membranes, especially affecting some body structures, such as the ocular surface. Specifically, the lipids of the corneal ocular surface present modifications related to age, with increased epithelial permeability making the elderly more vulnerable to corneal injuries. Inflammation is one of the most common pathologies evident in ophthalmology, which can affect different eye structures. Non-steroidal anti-inflammatory drugs (NSAIDs), such as ibuprofen, are widely used to treat ocular inflammation. Therefore, a drug delivery system such as PLGA biodegradable polymeric nanoparticles is an interesting alternative, as it can improve drug bioavailability, reduce the number of administrations required, and increase therapeutic adherence. This work also encompassed the proposal of a methodology for the development of formulations of nanoparticles that contain encapsulated dexibuprofen, and the creation of a model of corneal membrane that would permit in vitro studies of interactions. Keywords: dexibuprofen, biological membranes, eye, nanoparticles

Usefulness of the Measurement of Serum Paraoxonase-1 Arylesterase Activity in the Diagnoses of COVID-19

The development of inexpensive, fast, and reliable screening tests for COVID-19 is, as yet, an unmet need. The present study was aimed at evaluating the usefulness of serum arylesterase activity of paraoxonase-1 (PON1) measurement as a screening test in patients with different severity levels of COVID-19 infection. We included 615 COVID-19-positive patients who were classified as asymptomatic, mildly symptomatic, severely symptomatic, or fatally symptomatic. Results were compared with 50 healthy volunteers, 330 patients with cancer, and 343 with morbid obesity. Results showed PON1 activity greatly decreased in COVID-19 compared to healthy volunteers; a receiver operating characteristics plot showed a high diagnostic accuracy. The degree of COVID-19 severity did not influence PON1 levels. Our results indicated that PON1 determination was efficient for disease diagnosis, but not for prognosis. Furthermore, patients with obesity or cancer presented alterations similar to those of COVID-19 patients. As such, elevated levels of PON1 indicate the absence of COVID-19, but low levels may be present in various other chronic diseases. The assay is fast and inexpensive. We suggest that PON1 measurement could be used as an initial, high cut-off point screening method, while lower values should be confirmed with the more expensive nucleic acid amplification test

Multi-strange baryon production in pp collisions at √s=7 TeV with ALICE

A measurement of the multi-strange Ξ− and Ω− baryons and their antiparticles by the ALICE experiment at the CERN Large Hadron Collider (LHC) is presented for inelastic proton–proton collisions at a centre-of-mass energy of 7 TeV. The transverse momentum (pT) distributions were studied at mid-rapidity (|y|6.0 GeV/c. We also illustrate the difference between the experimental data and model by comparing the corresponding ratios of (Ω−+Ω¯+)/(Ξ−+Ξ¯+) as a function of transverse mass

Inclusive J/ψ production in pp collisions at √s=2.76 TeV

The ALICE Collaboration has measured inclusive J/ψ production in pp collisions at a center-of-mass energy √s=2.76 TeV at the LHC. The results presented in this Letter refer to the rapidity ranges |y|<0.9 and 2.5<y<4 and have been obtained by measuring the electron and muon pair decay channels, respectively. The integrated luminosities for the two channels are Linte=1.1 nb−1 and Lintμ=19.9 nb−1, and the corresponding signal statistics are NJ/ψe+e−=59±14 and NJ/ψμ+μ−=1364±53. We present dσJ/ψ/dy for the two rapidity regions under study and, for the forward-y range, d2σJ/ψ/dydpt in the transverse momentum domain 0<pt<8 GeV/c. The results are compared with previously published results at s=7 TeV and with theoretical calculations

Ks0Ks0 correlations in pp collisions at √s=7 TeV from the LHC ALICE experiment

Identical neutral kaon pair correlations are measured in √s=7 TeV pp collisions in the ALICE experiment. One-dimensional Ks0Ks0 correlation functions in terms of the invariant momentum difference of kaon pairs are formed in two multiplicity and two transverse momentum ranges. The femtoscopic parameters for the radius and correlation strength of the kaon source are extracted. The fit includes quantum statistics and final-state interactions of the a0/f0 resonance. Ks0Ks0 correlations show an increase in radius for increasing multiplicity and a slight decrease in radius for increasing transverse mass, mT, as seen in ππ correlations in pp collisions and in heavy-ion collisions. Transverse mass scaling is observed between the Ks0Ks0 and ππ radii. Also, the first observation is made of the decay of the f2′(1525) meson into the Ks0Ks0 channel in pp collisions