Culture collection of freshwater microalgae from the Azores archipelago: resource for taxonomic and phycoprospecting research

ABSTRACT: In recent years, much interest has been focused on the biotechnological potential of microalgae, mainly due to their rapid growth and the identification of several substances synthesized by these organisms. The isolation and in vitro cultivation of native microalgae species is very important for taxonomic and conservation studies and is the first step towards feasibility studies of local commercial productions. In this study 114 isolates of microalgae were obtained from samples collected in 23 locations of the island of Sao Miguel, Azores. From them, 60 species were identified comprising 39 Chlorophyta, 10 Ochrophyta, 6 Cyanophyta, 3 Charophyta, 1 Euglenozoa and 1 Cryptophyta. Eighteen of the species identified constitute new records for the Island of Sao Miguel.info:eu-repo/semantics/publishedVersio

Layer-by-layer assembled cell instructive nanocoatings containing platelet lysate

Great efforts have been made to introduce growth factors (GFs) onto 2D/3D constructs in order to control cell behavior. Platelet Lysate (PL) presents itself as a cost-effective source of multiple GFs and other proteins. The instruction given by a construct-PL combination will depend on how its instructive cues are presented to the cells. The content, stability and conformation of the GFs affect their instruction. Strategies for a controlled incorporation of PL are needed. Herein, PL was incorporated into nanocoatings by layer-by-layer assembling with polysaccharides presenting different sulfation degrees (SD) and charges. Heparin and several marine polysaccharides were tested to evaluate their PL and GF incorporation capability. The consequent effects of those multilayers on human adipose derived stem cells (hASCs) were assessed in short-term cultures. Both nature of the polysaccharide and SD were important properties that influenced the adsorption of PL, vascular endothelial growth factor (VEGF), fibroblast growth factor b (FGFb) and platelet derived growth factor (PDGF). The sulfated polysaccharides-PL multilayers showed to be efficient in the promotion of morphological changes, serum-free adhesion and proliferation of high passage hASCs (P>5). These biomimetic multilayers promise to be versatile platforms to fabricate instructive devices allowing a tunable incorporation of PL.Portuguese Foundation for Science and Technology is gratefully acknowledged for fellowships of Sara M. Oliveira. (SFRH/BD/70107/2010).r The research leading to these results has received funding from the European Union's Seventh Framework Programme (FP7/2007-2013) under grant agreement no REGPOT-CT2012-316331-POLARIS and FP7-KBBE-2010-4-266033 - SPECIAL. This work was also supported by the European Research Council grant agreement ERC-2012-ADG 20120216-321266 for the project ComplexiTE. The authors acknowledge Rogerio P. Pirraco for the Flow cytometry analysis

Analysis of wind velocity and release angle effects on discus throw using computational fluid dynamics

The aim of this paper is to study the aerodynamics of discus throw. A comparison of numerical and experimental performance of discus throw with and without rotation was carried out using the analysis of lift and drag coefficients. Initial velocity corresponding to variation angle of around 35.5° was simulated. Boundary condition, on the top and bottom boundary edges of computational domain, was imposed in order to eliminate external influences on the discus; a wind resistance was calculated for the velocity values of 25 and 27 m/s. The results indicate that the flight distance (D) was strongly affected by the drag coefficient, the initial velocity, the release angle and the direction of wind velocity. It was observed that these variables change as a function of discus rotation. In this study, results indicate a good agreement of D between experimental values and numerical results.info:eu-repo/semantics/publishedVersio

Development of new chitosan/carrageenan nanoparticles for drug delivery applications

The use of polymeric nanoparticles, especially those composed of natural polymers, has become a very interesting approach in drug delivery., mainly because of the advantages offered by their small dimensions. The aim of this work was to develop a novel formulation of nanoparticles comprised of two natural marine-derived polymers, namely chitosan and carrageenan, and to evaluate their potential for the association and controlled release of macromolecules. Nanoparticles were obtained in a hydrophilic environment, under very mild conditions, avoiding the use of organic solvents or other aggressive technologies for their preparation. The developed nanocarriers presented sizes within 350-650 run and positive zeta potentials of 50-60 mV. Polymeric interactions between nanoparticles' components were evaluated by Fourier transform infrared spectroscopy. Using ovalbumin as model protein, nanoparticles evidenced loading capacity varying from 4% to 17%, and demonstrated excellent capacity to provide a controlled release for up to 3 weeks. Furthermore, nanoparticles have demonstrated to exhibit a noncytotoxic behavior in biological in vitro tests performed using L929 fibroblasts, which is critical regarding the biocompatibility of those carriers. In summary, the developed chitosan-carrageenan nanoparticles have shown promising properties to be used as carriers of therapeutic macromolecules, with potential application not only strictly in drug delivery, but also in broader areas, such as tissue engineering and regenerative medicine. (C) 2009 Wiley Periodicals, Inc. J Biomed Mater Res 92A: 1265-1272, 2010Contract grant sponsor: Portuguese Foundation for Science and Technology (FCT) (POCTI/FEDER programmes)Contract grant sponsor: European Union STREP Project HIPPOCRATES; contract grant number: NMP3-CT-2003505758Contract grant sponsor: European NoE EXPERTISSUES; contract grant number: NMP3-CT-2004-50028

A potential signature for muscle-invasive tumors

Funding: This work was supported by the Astellas European Foundation Uro-Oncology Grant 2014; JABA RECORDATI Urology 2011 - Portuguese Society of Urology; Portuguese Foundation for Science and Technology (Strategic Project: PEst-C/SAU/UI3282/2013 and UID/NEU/04539/2013); GAI/FMUC and Santander Totta (Ref. FreitasGomes04.01.13) and Center of Investigation in Environment, Genetics and Oncobiology (CIMAGO), Portugal (Ref. 14/12). Margarida FerreiraTeixeira benefits from a PhD scholarship from the Portuguese Foundation for Science and Technology (Reference SFRH/BD/77314/2011).Striking evidence associates cancer stem cells (CSCs) to the high recurrence rates and poor survival of patients with muscle-invasive bladder cancer (BC). However, the prognostic implication of those cells in risk stratification is not firmly established, mainly due to the functional and phenotypic heterogeneity of CSCs populations, as well as, to the conflicting data regarding their identification based on a single specific marker. This emphasizes the need to exploit putative CSC-related molecular markers with potential prognostic significance in BC patients. This study aimed to isolate and characterize bladder CSCs making use of different functional and molecular approaches. The data obtained provide strong evidence that muscle-invasive BC is enriched with a heterogeneous stem-like population characterized by enhanced chemoresistance and tumor initiating properties, able to recapitulate the heterogeneity of the original tumor. Additionally, a logistic regression analysis identified a 2-gene stem-like signature (SOX2 and ALDH2) that allows a 93% accurate discrimination between non-muscle-invasive and invasive tumors. Our findings suggest that a stemness-related gene signature, combined with a cluster of markers to more narrowly refine the CSC phenotype, could better identify BC patients that would benefit from a more aggressive therapeutic intervention targeting CSCs population.publishersversionpublishe

Assessment of the suitability of chitosan/polybutylene succinate scaffolds seeded with mouse mesenchymal progenitor cells for a cartilage tissue engineering approach

In this work, scaffolds derived from a new biomaterial originated from the combination of a natural material and a synthetic material were tested for assessing their suitability for cartilage tissue engineering applications. In order to obtain a better outcome result in terms of scaffolds’ overall properties, different blends of natural and synthetic materials were created. Chitosan and polybutylene succinate (CPBS) 50/50 (wt%) were melt blended using a twin-screw extruder and processed into 5 5 5mm scaffolds by compression moulding with salt leaching. Micro-computed tomography analysis calculated an average of 66.29% porosity and 92.78% interconnectivity degree for the presented scaffolds. The salt particles used ranged in size between 63 and 125 lm, retrieving an average pore size of 251.28 lm. Regarding the mechanical properties, the compressive modulus was of 1.73 ± 0.4MPa (Esec 1%). Cytotoxicity evaluation revealed that the leachables released by the developed porous structures were not harmful to the cells and hence were noncytotoxic. Direct contact assays were carried out using a mouse bone marrow–derived mesenchymal progenitor cell line (BMC9). Cells were seeded at a density of 5 105 cells/scaffold and allowed to grow for periods up to 3 weeks under chondrogenic differentiating conditions. Scanning electron microscopy analysis revealed that the cells were able to proliferate and colonize the scaffold structure, and MTS test demonstrated cell viability during the time of the experiment. Finally, Western blot performed for collagen type II, a natural cartilage extracellular matrix component, showed that this protein was being expressed by the end of 3 weeks, which seems to indicate that the BMC9 cells were being differentiated toward the chondrogenic pathway. These results indicate the adequacy of these newly developed C-PBS scaffolds for supporting cell growth and differentiation toward the chondrogenic pathway, suggesting that they should be considered for further studies in the cartilage tissue engineering field.J. T. Oliveira would like to acknowledge the grant (SFRH/ BD17135/2004) from Portuguese Foundation for Science and Technology (FCT). The authors would like to thank Fernanda Marques, at the Institute for Health and Life Sciences (ICVS), University of Minho, Braga, Portugal, for her help with the Western blot analysis, as well as the staff at ICVS for allowing to use their facilities. The monoclonal antibody for collagen type II was obtained from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the Department of Biological Sciences, University of Iowa (Iowa City, IA). This work was carried out under the scope of the European NoE EXPERTISSUES (NMP3-CT-2004-500283), and partially supported by the European Project GENOSTEM (LSHB-CT-2003-503161) and the FCT Project CartiScaff (POCTI/SAU-BMA/58991/2004)

Development and characterization of PHB-HV based 3D scaffolds for a tissue engineering and cell-therapy combinatorial approach for spinal cord Injury regeneration

Spinal cord injury (SCI) leads to devastating neurological deficits. Several tissue engineering (TE)- based approaches have been investigated for repairing this condition. Poly (3-hydroxybutyrateco- 3-hydroxyvalerate) (PHB-HV) is found to be particularly attractive for TE applications due to its properties, such as biodegradability, biocompatibility, thermoplasticity and piezoelectricity. Hence, this report addresses the development and characterization of PHB-HV-based 3D scaffolds, produced by freeze-drying, aimed to SCI treatment. The obtained scaffolds reveal an anisotropic morphology with a fully interconnected network of pores. In vitro studies demonstrate a lack of cytotoxic effect of PHB-HV scaffolds. Direct contact assays also reveal their ability to support the culture of CNS-derived cells and mesenchymal-like stem cells from different sources. Finally, histocompatibility studies show that PHB-HV scaffolds are well tolerated by the host tissue, and do not negatively impact the left hindlimb locomotor function recovery. Therefore results herein presented suggest that PHB-HV scaffolds may be suitable for SCI treatment.This study was supported by the Portuguese Foundation for Science and Technology (FCT; Grant no PTDC/SAU-BMA/114059/2009; PEst-C/SAU/LA0001/2013-2014 and RNEM-REDE/1506/REM/2005) and Foundation Calouste Gulbenkian, under the scope of the Gulbenkian Program to Support Cutting Edge Research in Life Sciences (A.J.S.). This work was also partially supported by the European FP7 Project Find and Bind (NMP4-SL-2009-229292). The authors would like to thank Miguel Carvalho, Fabio Teixeira, and Filipa Campos for their collaboration in in vivo experiments

OMICs approaches in diarrhetic shellfish toxins research

ReviewDiarrhetic shellfish toxins (DSTs) are among the most prevalent marine toxins in Europe’s and in other temperate coastal regions. These toxins are produced by several dinoflagellate species; however, the contamination of the marine trophic chain is often attributed to species of the genus Dinophysis. This group of toxins, constituted by okadaic acid (OA) and analogous molecules (dinophysistoxins, DTXs), are highly harmful to humans, causing severe poisoning symptoms caused by the ingestion of contaminated seafood. Knowledge on the mode of action and toxicology of OA and the chemical characterization and accumulation of DSTs in seafood species (bivalves, gastropods and crustaceans) has significantly contributed to understand the impacts of these toxins in humans. Considerable information is however missing, particularly at the molecular and metabolic levels involving toxin uptake, distribution, compartmentalization and biotransformation and the interaction of DSTs with aquatic organisms. Recent contributions to the knowledge of DSTs arise from transcriptomics and proteomics research. Indeed, OMICs constitute a research field dedicated to the systematic analysis on the organisms’ metabolisms. The methodologies used in OMICs are also highly e ective to identify critical metabolic pathways a ecting the physiology of the organisms. In this review, we analyze the main contributions provided so far by OMICs to DSTs research and discuss the prospects of OMICs with regard to the DSTs toxicology and the significance of these toxins to public health, food safety and aquacultureinfo:eu-repo/semantics/publishedVersio

Finely tuned fiber-based porous structures for bone tissue engineering applications

info:eu-repo/semantics/publishedVersio