A Computing and Storage Server Infrastructure for a Mobile Application

The expansion of digital photography has turned a once expensive task into an easily accessible activity. Particularly, the dissemination of smartphones gave us the ability of takingmultiple pictures at no cost, which was just not possible in the past with traditional film cameras. With the current tools, we can use our smartphones to capture moments that we wish to revisit or share with our friends. However, due to having no associated cost, we can easily take multiple photos of the same motive, ending up with several similar pictures. This makes it difficult to share them with our friends and peers, as it is bothersome to navigate through many identical photos. While a simple solution would be to simply delete the repeated photos, this is often a difficult process: how can we be sure which of the photos is the best one? For this reason, we end up keeping more than one photo, which perpetuates the issue. This work is part of a larger project that aims to solve this problem. The project is composed of a mobile app and a server infrastructure. Due to processing capabilities and energy restrictions, the mobile device off-loads the image pre-processing to the remote server infrastructure, in order to extract measurable technical features. With this work we plan to design and provide a server infrastructure that will be able to assist the mobile app (and the photographer) in the process of removing duplicate photos, by providing services that offer features like similar image detection, image scoring and easing the process of selecting the best one that shall be kept while the others may be safely removed. The server infrastructure shall handle requests efficiently, in order to allow users with large photo galleries to remove their duplicate photos

39. Modulation of Histone Dosage and Chromatin Dynamics Regulates AAV Transduction

Recombinant adeno-associated vectors (AAVs) have gained momentum due to their combined characteristics of safety and efficiency. Nonetheless, the identity of the host functions intercepting the AAV transduction pathway still needs to be thoroughly understood.To address this issue on a genome-wide scale, we previously performed an unbiased RNAi high throughput screening (HTS; 18,120 human target genes) and identified 710 negative and 414 positive regulators of AAV efficiency. Based on the results obtained and with the purpose to identify factors involved in single-stranded (ss) AAV genome processing, we compared, again in an HTS format, siRNAs affecting ssAAV, but dispensable for scAAV transduction. One of the genes identified was ERI-1. While the effects of this protein were negligible on scAAV transduction, its knock down or overexpression lead to a 5 fold decrease or 2-7 fold increase of ssAAV efficiency, respectively.ERI-1 is a 3'-exoribonuclease known to degrade endogenous miRNAs and histone mRNAs. Indeed, we observed that AAV transduction negatively correlated with histone mRNA levels. Chromatin immunoprecipitation (ChIP) studies aimed at assessing the extent of chromatinization of the AAV genome revealed that the overexpression of ERI-1 determined an over 10-fold, selective reduction on ssAAV genome association with H3 and H4, while changes were negligible for scAAV DNA and for control cellular genes. Consistent with chromatin exerting a repressive role on ssAAV transduction, we also noticed that the downregulation of the main replication-dependent histone chaperone CAF-1 induced an over 20-fold increase in transduction. Increase of ssAAV2 transduction by ERI-1 also decreased the association, with the viral DNA, of proteins of the cellular DNA damage response (DDR; e.g. Nbs1 and Mre11), which our previous work had indicated as inhibitory of AAV transduction.Interestingly, DNA damage per se induced downregulation of histone gene expression. In particular, hydroxyurea, a drug markedly increasing AAV transduction, also determined histone mRNA degradation, an effect that required integrity of ERI-1.These results underline the importance of chromatin and its dynamic regulation in determining the fate of productive AAV transduction. These findings can be exploited for the development of more effective AAV-mediated gene delivery strategies

Efeito da maturidade, antropometria, composição corporal e potência dos membros inferiores sobre a dor reportada por atletas de futebol ao longo de uma época competitiva.

Mestrado em Exercício e SaúdeO objetivo deste estudo foi a identificação dos fatores (potência muscular, maturidade, antropometria e composição corporal) que mais se relacionam com a dor nos membros inferiores (MI) durante uma época competitiva em jovens futebolistas. A amostra compreendeu 76 atletas da Escola de Futebol Carlos Queiróz avaliados em dois momentos (novembro e maio), com uma idade média de 12,7 ± 1,7 anos e de 13,1 ± 1,7 anos, respetivamente no 1º (1M) e 2º momento de avaliação (2M). As variáveis antropométricas foram obtidas de acordo com o ISAK (Marfell-Jones et al. 2006), a composição corporal através das equações propostas por Frisancho (2008), a potência dos MI através do salto sem e com contramovimento e a maturidade através das equações de Mirwald et al. (2002). Verificámos que entre 1M e 2M a intensidade da dor aumentou significativamente (p≤0,012) na região anca-coxa (AC), e apenas não existiram diferenças significativas para as variáveis: salto sem contra movimento, IMC, alturas trocantérica e tibial lateral relativa, soma de pregas relativa e área muscular da coxa. No entanto, a maior parte das diferenças registadas resultaram da diferença de maturidade. Concluímos que 14,3% da variabilidade da dor AC no 2M foi explicada pela soma de pregas relativa e pela diferença altura tibial relativa

Genome-wide RNAi screening identifies host restriction factors critical for in vivo AAV transduction

Viral vectors based on the adeno-associated virus (AAV) hold great promise for in vivo gene transfer; several unknowns, however, still limit the vectors' broader and more efficient application. Here, we report the results of a high-throughput, whole-genome siRNA screening aimed at identifying cellular factors regulating AAV transduction. We identified 1,483 genes affecting vector efficiency more than 4-fold and up to 50-fold, either negatively or positively. Most of these factors have not previously been associated to AAV infection. The most effective siRNAs were independent from the virus serotype or analyzed cell type and were equally evident for single-stranded and self-complementary AAV vectors. A common characteristic of the most effective siRNAs was the induction of cellular DNA damage and activation of a cell cycle checkpoint. This information can be exploited for the development of more efficient AAV-based gene delivery procedures. Administration of the most effective siRNAs identified by the screening to the liver significantly improved in vivo AAV transduction efficiency

Glucocorticoid receptor signalling activates YAP in breast cancer

The Hippo pathway is an oncosuppressor signalling cascade that plays a major role in the control of cell growth, tissue homoeostasis and organ size. Dysregulation of the Hippo pathway leads to aberrant activation of the transcription co-activator YAP (Yes-associated protein) that contributes to tumorigenesis in several tissues. Here we identify glucocorticoids (GCs) as hormonal activators of YAP. Stimulation of glucocorticoid receptor (GR) leads to increase of YAP protein levels, nuclear accumulation and transcriptional activity in vitro and in vivo. Mechanistically, we find that GCs increase expression and deposition of fibronectin leading to the focal adhesion-Src pathway stimulation, cytoskeleton-dependent YAP activation and expansion of chemoresistant cancer stem cells. GR activation correlates with YAP activity in human breast cancer and predicts bad prognosis in the basal-like subtype. Our results unveil a novel mechanism of YAP activation in cancer and open the possibility to target GR to prevent cancer stem cells self-renewal and chemoresistance

In Vivo High-Content Evaluation of Three-Dimensional Scaffolds Biocompatibility

While developing tissue engineering strategies, inflammatory response caused by biomaterials is an unavoidable aspect to be taken into consideration, as it may be an early limiting step of tissue regeneration approaches. We demonstrate the application of flat and flexible films exhibiting patterned high-contrast wettability regions as implantable platforms for the high-content in vivo study of inflammatory response caused by biomaterials. Screening biomaterials by using high-throughput platforms is a powerful method to detect hit spots with promising properties and to exclude uninteresting conditions for targeted applications. High-content analysis of biomaterials has been mostly restricted to in vitro tests where crucial information is lost, as in vivo environment is highly complex. Conventional biomaterials implantation requires the use of high numbers of animals, leading to ethical questions and costly experimentation. Inflammatory response of biomaterials has also been highly neglected in high-throughput studies. We designed an array of 36 combinations of biomaterials based on an initial library of four polysaccharides. Biomaterials were dispensed onto biomimetic superhydrophobic platforms with wettable regions and processed as freeze-dried three-dimensional scaffolds with a high control of the array configuration. These chips were afterward implanted subcutaneously in Wistar rats. Lymphocyte recruitment and activated macrophages were studied on-chip, by performing immunocytochemistry in the miniaturized biomaterials after 24 h and 7 days of implantation. Histological cuts of the surrounding tissue of the implants were also analyzed. Localized and independent inflammatory responses were detected. The integration of these data with control data proved that these chips are robust platforms for the rapid screening of early-stage in vivo biomaterials’ response

Recent advances on open fluidic systems for biomedical applications: A review

Microfluidics has become an important tool to engineer microenvironments with high precision, comprising devices and methods for controlling and manipulating fluids at the submillimeter scale. A specific branch of microfluidics comprises open fluidic systems, which is mainly characterized by displaying a higher air/liquid interface when compared with traditional closed-channel setups. The use of open channel systems has enabled the design of singular architectures in devices that are simple to fabricate and to clean. Enhanced functionality and accessibility for liquid handling are additional advantages inputted to technologies based on open fluidics. While benchmarked against closed fluidics approaches, the use of directly accessible channels decreases the risk of clogging and bubble-driven flow perturbation. In this review, we discuss the advantages of open fluidics systems when compared to their closed fluidics counterparts. Platforms are analyzed in two separated groups based on different confinement principles: wall-based physical confinement and wettability-contrast confinement. The physical confinement group comprises both open and traditional microfluidics; examples based on open channels with rectangular and triangular cross-section, suspended microfluidics, and the use of narrow edge of a solid surface for fluid confinement are addressed. The second group covers (super)hydrophilic/(super)hydrophobic patterned surfaces, and examples based on polymer-, textile- and paper-based microfluidic devices are explored. The technologies described in this review are critically discussed concerning devices' performance and versatility, manufacturing techniques and fluid transport/manipulation methods. A gather-up of recent biomedical applications of open fluidics devices is also presented.European Research Council grant agreement ERC-2012-ADG 20120216-321266 for project ComplexiTE and ERC-2014-ADG-669858 for project “ATLAS”. N. M. Oliveira acknowledges the financial support from Portuguese Foundation for Science and Technology − FCT (Grant SFRH/BD/73172/2010), from the financial program POPH/FSE from QREN. The work was developed within the scope of the project CICECO Aveiro Institute of Materials, POCI-01-0145-FEDER-007679 (FCT Ref. UID/CTM/50011/2013). Sara Vilabril acknowledges the financial support from national funds through the FCT/MEC and when appropriate co-financed by FEDER under the PT2020 Partnership Agreementinfo:eu-repo/semantics/publishedVersio

Screening of perfused combinatorial 3D microenvironments for cell culture

Biomaterials combining biochemical and biophysical cues to establish close-to-extracellular matrix (ECM) models have been explored for cell expansion and differentiation purposes. Multivariate arrays are used as material-saving and rapid-to-analyze platforms, which enable selecting hit-spotted formulations targeting specific cellular responses. However, these systems often lack the ability to emulate dynamic mechanical aspects that occur in specific biological milieus and affect physiological phenomena including stem cells differentiation, tumor progression, or matrix modulation. We report a tailor-made strategy to address the combined effect of flow and biochemical composition of three-dimensional (3D) biomaterials on cellular response. We suggest a simple-to-implement device comprising (i) a perforated platform accommodating miniaturized 3D biomaterials and (ii) a bioreactor that enables the incorporation of the biomaterial-containing array into a disposable perfusion chamber. The system was upscaled to parallelizable setups, increasing the number of analyzed platforms per independent experiment. As a proof-of-concept, porous chitosan scaffolds with 1 mm diameter were functionalized with combinations of 5 ECM and cell-cell contact-mediating proteins, relevant for bone and dental regeneration, corresponding to 32 protein combinatorial formulations. Mesenchymal stem cells adhesion and production of an early osteogenic marker were assessed on-chip on static and under-flow dynamic perfusion conditions. Different hit-spotted biomaterial formulations were detected for the different flow regimes using direct image analysis. Cell-binding proteins still poorly explored as biomaterials components amelogenin and E-cadherin - were here shown as relevant cell response modulators. Their combination with ECM cell-binding proteins - fibronectin, vitronectin, and type 1 collagen - rendered specific biomaterial combinations with high cell adhesion and ALP production under flow. The developed versatile system may be targeted at wM.B. Oliveira acknowledges the financial support from Portuguese Foundation for Science and Technology- FCT (Grant SFRH/BPD/111354/2015). This work was developed within the scope of the projects CICECO-Aveiro Institute of Materials, POCI-01-0145-FEDER-007679 (FCT Ref. UID/CTM/50011/2013) and IPC/i3N Minho (FCT Ref. UID/CTM/50025/2013), financed by national funds through the FCT/MEC and when appropriate co-financed by FEDER under the PT2020 Partnership Agreement. This work was also supported by European Research Council grant agreement ERC-2014-ADG-669858 (project ATLAS)

Bioinspired multilayer membranes as potential adhesive patches for skin wound healing

Bioinspired and adhesive multilayer membranes are produced using the layer-by-layer (LbL) assembly of chitosan (CHT), alginate (ALG) and hyaluronic acid modified with dopamine (HA-DN). Freestanding multilayer membranes without DN are also produced as a control. The success of the synthesis of HA-DN was confirmed using UV-visible spectroscopy. Scanning electron microscopy images indicate that the surface of the DN-containing membranes is more porous than the control ones; they also present a higher average thickness value for the same number of CHT/ALG/CHT/HA(-DN) tetralayers (n = 100). Also, water uptake, mechanical strength and adhesion are enhanced with the introduction of DN moieties along the nano-layers. Besides, human dermal fibroblast viability, enhanced adhesion and proliferation were confirmed by immunofluorescence assays and by measuring both the metabolic activity and DNA content. Moreover, in vivo assays with such kinds of DN-containing multilayer membranes were performed; the application of these membranes in the treatment of dermal wounds induced in Wistar rats results in the highest decrease of inflammation of rat skin, compared with the control conditions. Overall, this investigation suggests that these mussel-inspired freestanding multilayer membranes may enhance either their mechanical performance or cellular adhesion and proliferation, leading to an improved wound healing process, being a promising material to restore the structural and functional properties of wounded skin.M. P. S. acknowledges the Portuguese Foundation for Science and Technology (FCT) for financial support through Grant No. SFRH/BD/97606/2013. This work was supported by the European Research Council grant agreement ERC-2014-ADG-669858 for the ATLAS project. The authors acknowledge Paula Marques (Mechanical Engineering Department, University of Aveiro, Aveiro, Portugal) for providing the mechanical equipment to carry out the tensile tests and the lap shear adhesion tests.info:eu-repo/semantics/publishedVersio