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Marine macroalgae or seaweeds synthesize a wide variety of polymers and smaller compounds with several bioactivities, among which the sulfated polysaccharides acquire greater relevance not only due to the reported antioxidant, antiviral and anticancer[1]Â activities, but also to the resemblance of extracellular matrix glycosaminoglycans found in the human body[2]. In this study, the potential of fucoidan (Fu) isolated from brown seaweed Fucus vesiculosus for therapeutical use has been evaluated, focusing in its performance as antitumoral agent (bioactive role) or as building block of cell encapsulating systems (structural role). Materials and Methods:Â The anticancer activity of Fu extracts was assessed by evaluating the cytotoxic behavior over two human breast cancer cell lines (MCF-7 and MDA-MB-231) in in-vitro culture, using human fibroblasts and endothelial cells (HPMEC-ST1 and MRC-5, respectively) as reference. Regarding the structural role, Fu was modified by methacrylation reaction (MFu) using methacrylic acid and further crosslinked using visible radiation and triethanolamine and eosin-y as photoinitiators. The photocrosslinking was performed on MFu solution droplets placed in a silica-based superhydrophobic surface[3], allowing the formation of particles[4]Â (since natural Fu is highly soluble in water and ion gelation is not effective). Biological performance of the developed particles was assessed by in vitro culture of fibroblasts and pancreatic cells (L929 and 1.1B4, respectively) in contact with MFu particles, up to 7 days. The ability of the developed materials to support adhesion and proliferation of cells was evaluated for both types of cells. Results and Discussion:Â The tested anticancer activity is not ubiquitous on Fu extracts, being dependent on its chemical features, with molecular weight (Mw) representing a particular role. Specifically, Mw values around 60 kDa exhibited cytotoxic effects to human breast cancer cell lines, while not affecting normal fibroblasts or endothelial cells (which represent the cells of the healthy tissue that would be closer to the tumor in a real situation). A concentration range of 0.2 to 0.3 mg mL-1 from the selected Fu extract could be considered as the therapeutic window for further studies. Regarding fucoidanâ s role on innovative biomaterials, the developed MFu particles could support the proliferation of fibroblasts (L929), but also of human pancreatic beta cells (1.1B4), which tend to form pseudo-islets after 7 days in culture (Fig. 1). This pancreatic cells could be also successfully encapsulated, opening a new route for a diabetes mellitus type 1 therapeutic approach. Fig. 1:Â Confocal microscopy images of 1.1B4 cells cultured in the presence of fucoidan-based particles and organized in pseudo-islets (red â actin; blue â nuclei). Conclusion:Â The present work establishes fucoidan as a high performance building block for the development of advanced therapies for cancer (targeted therapy) or tissue and organ regeneration. It shed light on the relation between chemical structure and biological activity towards anti-cancer effect and proposes novel beta cell laden particles as injectable insulin producing systems to tackle diabetes., Funding from projects 0687_NOVOMAR_1_P (co-funded by INTERREG 2007-2013 / POCTEP), CarbPol_u_Algae (EXPL/MAR-BIO/0165/2013, funded by the Portuguese Foundation for Science and Technology, FCT), POLARIS (FP7-REGPOT-CT2012-316331) and ComplexiTE (ERC-2012-ADG 20120216-321266), funded by the European Union’s Seventh Framework Programme for Research and Development is acknowledged. ASF, SSS, NMO and DSC are also thankful to FCT for their individual fellowships.

Aquaporins (AQPs), members of the water-channel protein family, are highly expressed in brain tissue especially in astrocytic end-feet. They are important players for water hemostasis during development of cytotoxic as well as vasogenic edema. Increased expression of AQPs is important in pathophysiology of neurological diseases such as neuroinflammation and ischemia. Unfortunately, there are a few pharmacological inhibitors of AQP4 with several side effects limiting their translation as a drug for use in clinical conditions. Another therapeutic approach is using antisense oligonucleotides (ASOs) to block AQP4 activity. These are short, synthetic, modified nucleic acids that bind RNA to modulate its function. However, they cannot pass the blood brain barrier (BBB). To overcome this obstacle we designed a nanoparticulate system made up of chitosan nanoparticles surface modified with PEG and conjugated with monoclonal anti transferrin receptor-1 antibody via streptavidin-biotin binding. The nanocarrier system could be targeted to the transferrin receptor-1 at the brain endothelial capillaries through monoclonal antibodies. It is hypothesized that the nanoparticles could pass the BBB via receptor mediated transcytosis and reach brain parenchyma. Particle size, zeta potential, loading capacity and release profiles of nanoparticles were investigated. It was observed that all types of chitosau (CS) nanoparticles had positive zeta potential values and nanoparticle particle size distribution varied between 100 and 800 nm. The association efficiency of ASOs into the nanoparticles was between 80–97% and the release profiles of the nanoparticles exhibited an initial burst effect followed by a controlled release. The results showed that the designed chitosan based nanocarriers could be a promising carrier system to transport nucleic acid based drugs to brain parenchyma This study is supported by The Scientific and Technological Research Council of Turkey (TUBITAK, Project Number: 110S460) SI

Publicado em "Journal of Tissue Engeneering and Regenerative Medicine", vol. 7, supp. 1 (2013), Glycosaminoglycans (GAGs) are integral part of the closest cellular environment: they can be found on the cells surface and in the extracellular matrix, where they interact with different proteins acting as local regulator of their activity. The use of GAGs in the preparation of protein delivery nanosystems is, therefore, prominent but so far, underexploited mainly because of the heterogeneity (composition and molecular weights) of natural glycans and the multistep procedures needed to obtain GAGs’ synthetic analogues and diblock copolymers.1 Recently, we have shown that oxime click reaction can be applied as a straightforward methodology for the synthesis of poly(ethylene glycol) (PEG)- hyaluronic acid (HA) diblock copolymers.2 These copolymers formed nanosized interpolyelectrolyte complexes (45 to 150 nm) by interaction with poly- L -lysine (PLL).3 Unfortunately, these complexes are not stable at physiological ionic strength. Herein, we describe a strategy to overcome this drawback; chondroitin sulphate-b-PEG diblock copolymers (CS-b-PEG) were obtained using the same oxime click reaction. The stronger negative charge of sulphate groups (versus the carboxilic groups present in HA) resulted in the complexes with higher stability: interpolyelectrolyte complexes between PLL and (CS-b-PEG) are stable up to 260 mM ionic strenght. Because carbohydrates do not activate Tcells, we believe that the reported herein complexes have an enormous potential in both drug delivery and vaccination fields.

Alpha-phenyl-n-tert-butyl nitrone (PBN) shows its major effect by scavenging free radicals formed in the ischemia and it has the ability to penetrate through the blood brain barrier easily. The in vivo stability of PBN is very low and when administered systemically, it has a mean plasma half life of about three hours. Therefore, formulations which are able to prolong the plasma residence time of PBN are of major interest, because oxygen radicals are usually continuously formed under pathological conditions. In this study, PBN, a nitrone compound having neuroprotective properties, was encapsulated in chitosan (CS) and chitosan-poly(ethylene glycol) (CS-PEG) nanoparticles for treatment of diseases such as stroke, in which sustained free radical production is reported. The nanoparticles were characterized through particle size determination, zeta potential, encapsulation efficiency, surface morphology determinations and in vitro release studies. The surface morphologies were evaluated by transmission electron microscopy (TEM) and nanoparticles having spherical shapes were characterized. The particle size distribution was between approximately 97 nm and approximately 322 nm; and the zeta potentials varied between approximately 9 mV and approximately 33 mV. Size of the nanoparticle formulations was important for the release of PBN from nanoparticles. The quantitative determination of PBN has been evaluated by a validated analytical HPLC method. The presented chitosan-based nanotechnology opens new perspectives for testing antioxidant activity in vivo.

Se mencionan 25 especies de líquenes del Parque Natural “Serra da Enciña da Lastra”, de las que son nuevas citas para Galicia: Acarospora murorum, Cetraria merrillii, Lecanora crenulata, Lecidella patavina, Placynthiella oligotropha, Placynthium lismorense, Scoliciosporum chlorococcum, Trapeliopsis wallrothii, Verrucaria calciseda y Verrucaria lecideoides. Las otras 15 son nuevas citas para la provincia de Ourense.

Presentamos los resultados de la revisión de los 116 pliegos de biotipo foliáceo de la familia Physciaceae depositados en el herbario PO de la Universidad de Porto, Portugal. Esta revisión nos permitió sinonimizar Physcia caesia var. perrugosa a P. aipolia, aportar la cita de Physconia grisea ssp. lilacina para Portugal y confirmar la única cita de Phaeophyscia sciastra.

Se mencionan 15 especies de líquenes del Parque Natural “Serra da Enciña da Lastra”, de las que 6: Fulgensia fulgens (Sw.) Elenkin, Melanelixia glabra (Schaer.) O. Blanco, A. Crespo, Divakar, Essl., D. Hawksw. & Lumbsch, Phaeophyscia sciastra (Ach.) Moberg, Psora decipiens (Hedw.) Hoffm., Psora testacea Hoffm. y Psora vallesiaca (Schaerer) Timdal son nuevas citas para Galicia. Las 9 restantes lo son para la provincia de Ourense.

Se señala la presencia en el Macizo orensano de 215 especies liquénicas, 21 de las cuales son desconocidas en el resto del territorio gallego. En esta zona se presenta también un gran número de taxones amenazados. Por ello, consideramos el LIC una importante área de conservación de la diversidad liquénica en Galicia.

A traditional karyomorphological approach was carried out in order to investigate the cytotaxonomic relationships among the species of the Santolina chamaecyparissus complex. All the 14 species included in this species complex were analysed for a total of 39 populations. Metaphase plates were obtained by squashing root tips stained with Feulgen technique. Measurable metaphase plates available in literature were also re-analysed. Chromosome number (2n), total haploid (monoploid) length (THL), and karyotype asymmetry indices (CVCL and MCA) were calculated for each of the 14 species. Chromosome data are provided for the first time for S. benthamiana and S. vedranensis, while a new hexaploid cytotype is reported for S. villosa. Five different ploidy levels are detected: diploid, triploid (a single individual of the otherwise diploid S. virens), tetraploid, pentaploid, and hexaploid. Our analyses reveal that polyploids are characterized by a more asymmetric karyotype with respect to diploids. The possible origin of polyploids, and in particular of the cultivated pentaploid S. chamaecyparissus, is briefly discussed.