Acetylcholine release and choline uptake by cuttlefish (Sepia officinalis) optic lobe synaptosomes

Acetylcholine (ACh), which is synthesized from choline (Ch), is believed to hold a central place in signaling mechanisms within the central nervous system (CNS) of cuttlefish (Sepia officinalis) and other coleoid cephalopods. Although the main elements required for cholinergic function have been identified in cephalopods, the transmembrane translocation events promoting the release of ACh and the uptake of Ch remain largely unsolved. The ACh release and Ch uptake were quantitatively studied through the use of in vitro chemiluminescence and isotopic methods on a subcellular fraction enriched in synaptic nerve endings (synaptosomes) isolated from cuttlefish optic lobe. The ACh release evoked by K+ depolarization was found to be very high (0.04 pmol ACh.s-(1).mg(-1) protein). In response to stimulation by veratridine, a secretagogue (a substance that induces secretion) that targets voltage-gated Na+ channels, the release rate and the total amount of ACh released were significantly lower, by 10-fold, than the response induced by KCl. The high-affinity uptake of choline was also very high (31 pmol Ch.min(-1).mg(-1) protein). The observed ACh release and Ch uptake patterns are in good agreement with published data on preparations characterized by high levels of ACh metabolism, adding further evidence that ACh acts as a neurotransmitter in cuttlefish optic lobe.FCTPOCTI/BSE/46721/2002SFRH/BPD/14677/2003SFRH/BD/1079/2000SFRH/BD/6403/2001SFRH/BD/18101/2004EC (to Y.D.) - LIPIDIET - QLK1-CT-2002-0017

Integrated biomimetic carbon nanotube composites for in vivo systems

CESAMAs interest in using carbon nanotubes for developing biologically compatible systems continues to grow, biological inspiration is stimulating new directions for in vivo approaches. The ability to integrate nanotechnology-based systems in the body will provide greater successes if the implanted material is made to mimic elements of the biological milieu especially through tuning physical and chemical characteristics. Here, we demonstrate the highly successful capacity for in vivo implantation of a new carbon nanotube-based composite that is, itself, integrated with a hydroxyapatite-polymethyl methacrylate to create a nanocomposite. The success of this approach is grounded in finely tailoring the physical and chemical properties of this composite for the critical demands of biological integration. This is accomplished through controlling the surface modification scheme, which affects the interactions between carbon nanotubes and the hydroxyapatite-polymethyl methacrylate. Furthermore, we carefully examine cellular response with respect to adhesion and proliferation to examine in vitro compatibility capacity. Our results indicate that this new composite accelerates cell maturation through providing a mechanically competent bone matrix; this likely facilitates osteointegration in vivo. We believe that these results will have applications in a diversity of areas including carbon nanotube, regeneration, chemistry, and engineering research.NANO/NMed-AT/0115/2007SFRH/BPD/14677/2003FC

Does neurotransmission impairment accompany aluminium neurotoxicity?

Neurobehavioral disorders, except their most overt form, tend to lie beyond the reach of clinicians. Presently, the use of molecular data in the decision-making processes is limited. However, as details of the mechanisms of neurotoxic action of aluminium become clearer, a more complete picture of possible molecular targets of aluminium can be anticipated, which promises better prediction of the neurotoxicological potential of aluminium exposure. In practical terms, a critical analysis of current data on the effects of aluminium on neurotransmission can be of great benefit due to the rapidly expanding knowledge of the neurotoxicological potential of aluminium. This review concludes that impairment of neurotransmission is a strong predictor of outcome in neurobehavioral disorders. Key questions and challenges for future research into aluminium neurotoxicity are also identified. (c) 2007 Elsevier Inc. All rights reserved.FCT (to V.S.S.)SFRH/BPD/14677/200

Comparative effects of aluminum and ouabain on synaptosomal choline uptake, acetylcholine release and (Na+/K+)ATPase

Closing the gap between adverse health effects of aluminum and its mechanisms of action still represents a huge challenge. Cholinergic dysfunction has been implicated in neuronal injury induced by aluminum. Previously reported data also indicate that in vivo and in vitro exposure to aluminum inhibits the mammalian (Na+/K+)ATPase, an ubiquitous plasma membrane pump. This study was undertaken with the specific aim of determining whether in vitro exposure to AlCl3 and ouabain, the foremost utilized selective inhibitor of (Na+/K+)ATPase, induce similar functional modifications of cholinergic presynaptic nerve terminals, by comparing their effects on choline uptake, acetylcholine release and (Na+/K+)ATPase activity, on subcellular fractions enriched in synaptic nerve endings isolated from rat brain, cuttlefish optic lobe and torpedo electric organ. Results obtained show that choline uptake by rat synaptosomes was inhibited by submillimolar AlCl3, whereas the amount of choline taken up by synaptosomes isolated from cuttlefish and torpedo remained unchanged. Conversely, choline uptake was reduced by ouabain to a large extent in all synaptosomal preparations analyzed. In contrast to ouabain, which modified the K+ depolarization evoked release of acetylcholine by rat, cuttlefish and torpedo synaptosomal fractions, AlCl3 induced reduction of stimulated acetylcholine release was only observed when rat synaptosomes were challenged. Finally, it was observed that the aluminum effect on cuttlefish and torpedo synaptosomal (Na+/K+)ATPase activity was slight when compared to its inhibitory action on mammalian (Na+/K+)ATPase. In conclusion, inhibition of (Na+/K+)ATPase by AlCl3 and ouabain jeopardized the high-affinity (Na+-dependent, hemicholinium-3 sensitive) uptake of choline and the Ca2+-dependent, K+ depolarization evoked release of acetylcholine by rat, cuttlefish and torpedo synaptosomal fractions. The effects of submillimolar AlCl3 On choline uptake and acetylcholine release only resembled those of ouabain when rat synaptosomes were assayed. Therefore, important differences were found between the species regarding the cholinotoxic action of aluminum. The variability of (Na+/K+)ATPase sensitivity to aluminum of cholinergic neurons might contribute to their differential susceptibility to this neurotoxic agent. (C) 2007 Elsevier Ireland Ltd. All rights reserved.FCTPOCTI/BSE/46721/2002SFRH/BPD/14677/2003SFRH/BD/1079/2000SFRH/BD/6403/2001SFRH/BD/18101/2004EC (to Y.D.)LIPIDIET - QLK1-CT-2002-0017

Aqueous Exfoliation of Transition Metal Dichalcogenides Assisted by DNA/RNA Nucleotides: Catalytically Active and Biocompatible Nanosheets Stabilized by Acid–Base Interactions

This document is the Accepted Manuscript version of a Published Work that appeared in final form in ACS Applied Materials & Interfaces, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://pubs.acs.org/doi/10.1021/acsami.6b13619The exfoliation and colloidal stabilization of layered transition metal dichalcogenides (TMDs) in an aqueous medium using functional biomolecules as dispersing agents have a number of potential benefits toward the production and practical use of the corresponding two-dimensional materials, but such a strategy has so far remained underexplored. Here, we report that DNA and RNA nucleotides are highly efficient dispersants in the preparation of stable aqueous suspensions of MoS2 and other TMD nanosheets at significant concentrations (up to 5–10 mg mL–1). Unlike the case of common surfactants, for which adsorption on 2D materials is generally based on weak dispersive forces, the exceptional colloidal stability of the TMD flakes was shown to rely on the presence of relatively strong, specific interactions of Lewis acid–base type between the DNA/RNA nucleotide molecules and the flakes. Moreover, the nucleotide-stabilized MoS2 nanosheets were shown to be efficient catalysts in the reduction of nitroarenes (4-nitrophenol and 4-nitroaniline), thus constituting an attractive alternative to the use of expensive heterogeneous catalysts based on noble metals, and exhibited an electrocatalytic activity toward the hydrogen evolution reaction that was not impaired by the possible presence of nucleotide molecules adsorbed on their active sites. The biocompatibility of these materials was also demonstrated on the basis of cell proliferation and viability assays. Overall, the present work opens new vistas on the colloidal stabilization of 2D materials based on specific interactions that could be useful toward different practical applications.Financial support from the Spanish Ministerio de Economía y Competitividad (MINECO) and the European Regional Development Fund (ERDF) through project MAT2015-69844-R is gratefully acknowledged. Partial funding by Plan de Ciencia, Tecnología e Innovación (PCTI) 2013-2017 del Principado de Asturias and the ERDF (project GRUPIN14-056) is also acknowledged. M.A-V. and J.M.M. are grateful to MINECO and the Spanish Ministerio de Educación, Cultura y Deporte (MECD), respectively, for their pre-doctoral contracts. O.P-V. acknowledges funding of his contract by the European Social Fund (ESF). M.C. is grateful to FCT (Fundação para a Ciência e a Tecnologia) for financial support through the post-doctoral grant SFRH/BPD/101468/2014, co-financed by national funds from the Portuguese Ministry of Education and Science and the ESF.Peer reviewe