Chitosan-Hyaluronic acid hybrid vectors for retinal gene therapy

info:eu-repo/semantics/publishedVersio

Sustained gene expression in the retina by improved episomal vectors

Gene and cellular therapies are nowadays part of therapeutic strategies for the treatment of diverse pathologies. The drawbacks associated with gene therapy-low levels of transgene expression, vector loss during mitosis, and gene silencing-need to be addressed. The pEPI-1 and pEPito family of vectors was developed to overcome these limitations. It contains a scaffold/matrix attachment region, which anchors its replication to cell division in eukaryotic cells while in an extrachromosomal state and is less prone to silencing, due to a lower number of CpG motifs. Recent success showed that ocular gene therapy is an important tool for the treatment of several diseases, pending the overcome of the aforementioned limitations. To achieve sustained gene delivery in the retina, we evaluated several vectors based on pEPito and pEPI-1 for their ability to sustain transgene expression in retinal cells. These vectors stably transfected and replicated in retinal pigment epithelial (RPE) cells. Expression levels were promoter dependent with constitutive promoters cytomegalovirus immediate early promoter (CMV) and human CMV enhancer/human elongation factor 1 alpha promoter yielding the highest levels of transgene expression compared with the retina-specific RPE65 promoter. When injected in C57Bl6 mice, transgene expression was sustained for at least 32 days. Furthermore, the retina-specific RPE65 promoter showed higher efficiency in vivo compared to in vitro. In this study, we demonstrate that by combining tissue-specific promoters with a mitotic stable system, less susceptible to epigenetic silencing such as pEPito-based plasmids, we can achieve prolonged gene expression and a sustained therapeutic effect.Fundacao para a Ciencia e Tecnologia, Portugal [PEst/OE/EQB-LA 0023/2013, SFRH/BD/76873/2011, SFRH/BD/70318/2010, PTDC/SAU/BEB/098475/2008]; European Union [PIRG-GA-2009-249314

An integrative approach for codon repeats evolutionary analyses

The relationship between genome characteristics and several human diseases has been a central research goal in genomics. Many studies have shown that specific gene patterns, such as amino acid repetitions, are associated with human diseases. However, several open questions still remain, such as, how these tandem repeats appeared in the evolutionary path or how they have evolved in orthologous genes of related organisms. In this paper, we present a computational solution that facilitates comparative studies of orthologous genes from various organisms. The application uses various web services to gather gene sequence information, local algorithms for tandem repeats identification and similarity measures for gene clustering.publishe

Development of new analytical tools for monitoring of cardiovascular disease markers – towards the detection of homocysteine-thiolactone

Poster presented at the 4th International Conference on Bio-Sensing Technology, 10-13 May 2015, Lisbon, Portuga

New PON1-based biosensor for the detection of homocysteine-thiolactone in human plasma

Poster presented at the XXIII International Symposium on Bioelectrochemistry and Bioenergetics, 14-18 June 2015, Malmo, Sweden

Computational and statistical methodologies for ORFeome primary structure analysis

Codon usage and context are biased in open reading frames (ORFs) of most genomes. Codon usage is largely influenced by biased genome G+C pressure, in particular in prokaryotes, but the general rules that govern the evolution of codon context remain largely elusive. In order to shed new light into this question we have developed computational, statistical and graphical tools for analysis of codon context on an ORFeome wide scale. In here, we describe these methodologies in detail and show how they can be used for analysis of ORFs of any genome sequenced.publishe

Graphene Oxide Thin Films with Drug Delivery Function

Funding Information: Funding: Funding was obtained through the Fundação para a Ciência e Tecnologia (FCT) through the projects iNOVA4Health, Translational Medicine program—UIDB/Multi/04462/2020; UIDB/50008/2020 and PTDC/CTM-REF/2679/2020.Graphene oxide has been used in different fields of nanomedicine as a manager of drug delivery due to its inherent physical and chemical properties that allow its use in thin films with biomedical applications. Several studies demonstrated its efficacy in the control of the amount and the timely delivery of drugs when it is incorporated in multilayer films. It has been demonstrated that oxide graphene layers are able to work as drug delivery or just to delay consecutive drug dosage, allowing the operation of time-controlled systems. This review presents the latest research developments of biomedical applications using graphene oxide as the main component of a drug delivery system, with focus on the production and characterization of films, in vitro and in vivo assays, main applications of graphene oxide biomedical devices, and its biocompatibility properties.publishersversionpublishe

Traffic light optimization of an intersection: a portuguese case study

Smart cities aim to rise strategies that reduce issues caused by the urban population growth and fast urbanization. Thus, traffic light optimization emerges as an important option for urban traffic management. The main goal of this study is to improve traffic light management at a specific intersection, in the City of Guimarães (Portugal), where high-intensity traffic and an active pedestrian area were observed, generating traffic queues. To achieve the goals, a simulation-based optimization strategy using the Particle Swarm Optimization combined with the Simulation of Urban Mobility software was used to minimize the average waiting time of the vehicles by determining the optimal value of the traffic light cycle. The computational results showed it is possible to decrease by 78.2% the average value of the waiting time. In conclusion, by better managing the traffic light cycle time, traffic flow without congestion or queues can be achieved.This work has been supported by FCT—Fundação para a Ciência e Tecnologia within the R&D Units Project Scope: UIDB/00319/2020 and the project “Integrated and Innovative Solutions for the well-being of people in complex urban centers” within the Project Scope NORTE-01-0145-FEDER-000086

The workaholism as an obstacle to safety and well-being in the workplace

The work aholism, defined as addiction at work, increase burnout and negatively affects the psychological well-being. It was developed a quantitative study using self-reported measures. This study aims to evaluate the variables: passion for work, work engagement, engagement for life and job satisfaction as predictors of workaholism and the effects of workaholism on the psychological well-being and burnout. With a sample of 199 workers, the results of multiple linear regression analysis allowed to identify that some of the variables and their dimensions have significant effects on the workaholism and on psychological well-being and burnout

Morphology dependence degradation of electro- and magnetoactive poly(3-hydroxybutyrate-co-hydroxyvalerate) for tissue engineering applications

Poly(hydroxybutyrate-co-hydroxyvalerate) (PHBV) is a piezoelectric biodegradable and biocompatible polymer suitable for tissue engineering applications. The incorporation of magnetostrictive cobalt ferrites (CFO) into PHBV matrix enables the production of magnetically responsive composites, which proved to be effective in the differentiation of a variety of cells and tissues. In this work, PHBV and PHBV with CFO nanoparticles were produced in the form of films, fibers and porous scaffolds and subjected to an experimental program allowing to evaluate the degradation process under biological conditions for a period up to 8 weeks. The morphology, physical, chemical and thermal properties were evaluated, together with the weight loss of the samples during the in vitro degradation assays. No major changes in the mentioned properties were found, thus proving its applicability for tissue engineering applications. Degradation was apparent from week 4 and onwards, leading to the conclusion that the degradation ratio of the material is suitable for a large range of tissue engineering applications. Further, it was found that the degradation of the samples maintain the biocompatibility of the materials for the pristine polymer, but can lead to cytotoxic effects when the magnetic CFO nanoparticles are exposed, being therefore needed, for magnetoactive applications, to substitute them by biocompatible ferrites, such as an iron oxide (Fe3O4).This work was supported by the Portuguese Foundation for Science and Technology (FCT) in the framework of the Strategic Funding UID/FIS/04650/2020, UID/BIO/04469/2020 and UID/QUI/00686/2020, and projects PTDC/BTM-MAT/28237/2017 and PTDC/EMD-EMD/28159/2017 and Associate Laboratory for Green Chemistry—LAQV, financed by national funds from FCT/MCTES (UIDB/50006/2020). The authors also thank the FCT for the SFRH/BPD/121526/2016 (DMC) grant. The authors acknowledge funding by the Spanish Ministry of Economy and Competitiveness (MINECO) through the project MAT2016-76039-C4-3-R (AEI/FEDER, UE) and from the Basque Government Industry and Education Departments under the ELKARTEK, HAZITEK and PIBA (PIBA-2018-06) programs, respectively.info:eu-repo/semantics/publishedVersio