Reply: Response to Wilson et al. Comments on Lopez-Jaramillo et al. DivinylSulfone Cross-Linked Cyclodextrin-Based Polymeric Materials: Synthesis and Applications as Sorbents and Encapsulating Agents. Molecules, 2015, 20, 3565–3581

Reply: Response to Wilson et al. Comments on Lopez-Jaramillo et al. DivinylSulfone Cross-Linked Cyclodextrin-Based Polymeric Materials: Synthesis and Applications as Sorbents and Encapsulating Agents. Molecules, 2015, 20, 3565–3581The authors acknowledge the Ministerio de Ciencia and Innovacion for financial support (Grant CTQ2011-29299-CO2-01)

Vinyl sulfone: a multi-purpose function in proteomics

The chapter is organized in three sections. In a first instance, a general overview of the vinyl sulfone chemistry in terms of the most relevant methods of synthesis and aspects of their reactivity will be followed by a discussion of the application of this chemical behaviour with proteins. Their advantages and disadvantages with other currently available methodologies to modify amine and thiol groups naturally present in proteins will be compared. In a second section the applications of vinyl sulfones to Proteomics will be enumerated. Finally, the wide scope of the vinyl sulfone chemistry in other omic sciences will be discussed.The authors acknowledge Dirección General de Investigación Científica y Técnica (DGICYT) (CTQ2008-01754) and Junta de Andalucía (P07-FQM-02899) for financial support

Turning carbon dots into selenium bearing nanoplatforms with in vitro GPx-like activity and pro-oxidant activity

Selenium (Se) has been defined as the “Janus element”, with one face showing antioxidant activity and the other pro-oxidant activity. The biological effect of Se depends on both dose and speciation. Se nanoparticles are attracting major interest, although their large-scale preparation for biomedical applications is not trivial. We hypothesize that acid anhydride-coated carbon dots (AACD) are an attractive platform for preparing nanoparticles containing chemically defined Se. The reaction of AA-CD with 3- selenocyanatopropan-1-amine yields carbon dots bearing selenocyanate and carboxylate groups (CD-SeCN) that allow for tuning the hydrosolubility. CD-SeCN has a Se content of 0.36 μmol per mg of nanoparticles, and they show the typical photoluminescence of carbon dots. The selenocyanate groups (SeCN) exhibited glutathione peroxidase-like activity and cytotoxicity. Data show that antioxidant behavior differs between normal and tumor cells, and the evaluation on HEK293 and A549 cells reveals that the toxicity of CD-SeCN depends on dose, time, and intracellular glutathione (GSH) content. The toxicity of CD-SeCN decreases with the time of incubation and the cell death mechanism switches from necrosis to apoptosis, indicating that CD-SeCN is neutralized. Additionally, high levels of intracellular GSH exert a protective effect. These results support a pharmacological potential in cancers with low levels of intracellular GSH. The use of AA-CD as nanoplatforms is a general strategy that paves the way for the engineering of advanced nanosystemsSpanish institution Ministerio de Ciencia, Innovacion y Universidades (No. CTQ2017- 86125-P)Unit of Excellence in Chemistry Applied to Biomedicine and the EnvironmentCentro de Instrumentacion Cientifica (Universidad de Granada)Universidad de Granada/CBUASpringer Natur

Sistema microcontrolado para el aprendizaje y la estimulación de discapacitados motóricos de corta edad (SAED)

En este trabajo se presenta un prototipo desarrollado, (dentro de la actividad investigadora entre la Asociación Tutelar del Minusválido de Cartagena –ASTUS- y el Grupo de Investigación “Electrónica Industrial y Médica”) a fin de facilitar el aprendizaje en el uso de pulsadores o conmutadores de acceso por parte de usuarios discapacitados. Así, a través del empleo de un microcontrolador, y haciendo uso de técnicas de depuración de accionamientos involuntarios de desarrollo propio, es posible adelantar la edad de iniciación de los discapacitados en el acceso a los sistemas informáticos, a través de juguetes estándar que actúen como elementos de estimulación lúdica

Vinyl sulfonyl chemistry-driven unidirectional transport of a macrocycle through a [2]rotaxane

This work has been financially supported by FEDER(EDRF)/Junta de Andalucia-Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades (P18-FR-2877), grant PID2020-112906GA-I00 funded by MCIN/AIE (/10.13039/501100011033) and Ministerio de Economia y Competitividad (MINECO, Spain) (CTQ2014-55474-C2-2-R and CTQ2017-86125P, co-financed by FEDER funds). Funding for open access APCs provided by Universidad de Granada through a Paid by Read & Publish agreement with RSC.By applying a combination of the coupling-and-decoupling (CAD) chemistry of the vinyl sulfonate group with the click thia-Michael addition to the vinyl sulfone group (MAVS) we performed the irreversible unidirectional transportation of the ring through the linear component in a [2]rotaxane by a chemically and pH-driven flashing energy ratchet mechanism. The design is based on a monostoppered thread precursor bearing a sulfonate stopper, a vinyl sulfone group on the unstoppered end and a dibenzylammonium unit as recognition site for the dibenzo-24-crown-8 macrocycle. First, the ring enters from the vinyl sulfone side and the rotaxane is capped through a thia-Michael addition reaction. Then, the cleavage of the sulfonate group of the opposite stopper using MgBr2 as chemical stimulus and subsequent addition of base (Et3N) promoted the controlled and directional release of the macrocycle into the bulk under mild conditions. The efficiency of the system allowed the in situ operation as demonstrated by NMR and HRMS techniques.FEDER(EDRF)/Junta de Andalucia-Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades - MCIN/AIE P18-FR-2877Ministerio de Economia y Competitividad (MINECO, Spain) (FEDER funds) CTQ2014-55474-C2-2-R; CTQ2017-86125PUniversidad de GranadaRSCMCIN/AIE PID2020-112906GA-I0

Single chain variable fragment fused to maltose binding protein: a modular nanocarrier platform for the targeted delivery of antitumorals

This work was supported by grants CTQ2014-55474-C2-1-R, CTQ2014-55474-C2-2-R and CTQ2017-86125-P from the Ministerio Economia, Industria y Competitividad (co-financed by FEDER funds). SP is supported by a FPU fellowship (FPU17/ 04749). We acknowledge the University of Granada (Spain) cell culture, animal and microscopy central facilities (CIC-UGR).The use of the specific binding properties of monoclonal antibody fragments such as single-chain variable fragments (ScFv) for the selective delivery of antitumor therapeutics for cancer cells is attractive due to their smaller size, low immunogenicity, and low-cost production. Although covalent strategies for the preparation of such ScFv-based therapeutic conjugates are prevalent, this approach is not straightforward, as it requires prior chemical activation and/or modification of both the ScFv and the therapeutics for the application of robust chemistries. A non-covalent alternative based on ScFv fused to maltose-binding protein (MBP) acting as a binding adapter is proposed for active targeted delivery. MBP-ScFv proves to be a valuable modular platform to synergistically bind maltose-derivatized therapeutic cargos through the MBP, while preserving the targeting competences provided by the ScFv. The methodology has been tested by using a mutated maltose-binding protein (MBP I334W) with an enhanced affinity toward maltose and an ScFv coding sequence toward the human epidermal growth factor receptor 2 (HER2). Non-covalent binding complexes of the resulting MBP-ScFv fusion protein with diverse maltosylated therapeutic cargos (a near-infrared dye, a maltosylated supramolecular beta-cyclodextrin container for doxorubicin, and non-viral polyplex gene vector) were easily prepared and characterized. In vitro and in vivo assays using cell lines that express or not the HER2 epitope, and mice xenografts of HER2 expressing cells demonstrated the capability and versatility of MBP-ScFv for diagnosis, imaging, and drug and plasmid active targeted tumor delivery. Remarkably, the modularity of the MBP-ScFv platform allows the flexible interchange of both the cargos and the coding sequence for the ScFv, allowing ad hoc solutions in targeting delivery without any further optimization since the MBP acts as a pivotal element.Ministerio Economia, Industria y Competitividad - FEDER funds CTQ2014-55474-C2-1-R CTQ2014-55474-C2-2-R CTQ2017-86125-PSpanish Government FPU17/0474

Divinyl Sulfone Cross-Linked Cyclodextrin-Based Polymeric Materials: Synthesis and Applications as Sorbents and Encapsulating Agents

The aim of this study was to evaluate the crosslinking abilities of divinyl sulfone (DVS) for the preparation of novel water-insoluble cyclodextrin-based polymers (CDPs) capable of forming inclusion complexes with different guest molecules. Reaction of DVS with native α-cyclodextrin (α-CD), β-cyclodextrin (β-CD) and/or starch generates a variety of homo- and hetero-CDPs with different degrees of crosslinking as a function of the reactants’ stoichiometric ratio. The novel materials were characterized by powder X-ray diffraction, electron microscopy and for their sorption of phenol and 4-nitrophenol. They were further evaluated as sorbents with phenolic pollutants (bisphenol A and β-naphthol) and bioactive compounds (the hormone progesterone and curcumin). Data obtained from the inclusion experiments show that the degree of cross-linking has a minor influence on the yield of inclusion complex formation and highlight the important role of the CDs, supporting a sorption process based on the formation of inclusion complexes. In general, the inclusion processes are better described by a Freundlich isotherm although an important number of them can also be fitted to the Langmuir isotherm with R2 ≥ 0.9, suggesting a sorption onto a monolayer of homogeneous sites

Poly(ethylene-imine)-Functionalized Magnetite Nanoparticles Derivatized with Folic Acid: Heating and Targeting Properties

Magnetite nanoparticles (MNPs) coated by branched poly (ethylene-imine) (PEI) were synthesized in a one-pot. Three molecular weights of PEI were tested, namely, 1.8 kDa (sample MNP-1), 10 kDa (sample MNP-2), and 25 kDa (sample MNP-3). The MNP-1 particles were further functionalized with folic acid (FA) (sample MNP-4). The four types of particles were found to behave magnetically as superparamagnetic, with MNP-1 showing the highest magnetization saturation. The particles were evaluated as possible hyperthermia agents by subjecting them to magnetic fields of 12 kA/m strength and frequencies ranging between 115 and 175 kHz. MNP-1 released the maximum heating power, reaching 330 W/g at the highest frequency, in the high side of reported values for spherical MNPs. In vitro cell viability assays of MNP-1 and MNP-4 against three cell lines expressing different levels of FA receptors (FR), namely, HEK (low expression), and HeLa (high expression), and HepG2 (high expression), demonstrated that they are not cytotoxic. When the cells were incubated in the presence of a 175 kHz magnetic field, a significant reduction in cell viability and clone formation was obtained for the high expressing FR cells incubated with MNP-4, suggesting that MNP-4 particles are good candidates for magnetic field hyperthermia and active targeting.Spanish Institutions: Ministerio de Ciencia, Innovación y Universidades (PGC2018-098770-B-I00 and CTQ2017-86125-P)Junta de Andalucía (ProgramaOperativo FEDER 2014-2020, grants B-FQM-141-UGR18, A1-FQM-341-UGR-18, C-FQM-497-UGR18

Vinyl sulfone silica: application of an open preactivated support to the study of transnitrosylation of plant proteins by S-nitrosoglutathione

Background S-nitrosylaton is implicated in the regulation of numerous signaling pathways with a diversity of regulatory roles. The high lability of the S-NO bond makes the study of proteins regulated by S-nitrosylation/denitrosylation a challenging task and most studies have focused on already S-nitrosylated proteins. We hypothesize that: i) S-nitrosoglutathione (GSNO) transnitrosylation is a feasible mechanism to account for the physiological S-nitrosylation of rather electropositive sulfur atoms from proteins, ii) affinity chromatography is a suitable approach to isolate proteins that are prone to undergo S-transnitrosylation and iii) vinyl sulfone silica is a suitable chromatographic bead.Results The combination of vinyl sulfone silica with GSNO yielded an affinity resin that withstood high ionic strength without shrinking or deforming and that it was suitable to isolate potential GSNO transnitrosylation target candidates. Fractions eluted at 1500 mM NaCl resulted in a symmetrical peak for both, protein and S-nitrosothiols, supporting the idea of transnitrosylation by GSNO as a selective process that involves strong and specific interactions with the target protein. Proteomic analysis led to the identification of 22 physiological significant enzymes that differ with the tissue analyzed, being regulatory proteins the most abundant group in hypocotyls. The identification of chloroplastidic FBPase, proteasome, GTP-binding protein, heat shock Hsp70, syntaxin, catalase I, thioredoxin peroxidase and cytochrome P450 that have already been reported as S-nitrosylated by other techniques can be considered as internal positive controls that validate our experimental approach. An additional validation was provided by the prediction of the S-nitrosylation sites in 19 of the GSNO transnitrosylation target candidates.Conclusions Vinyl sulfone silica is an open immobilization support that can be turned ad hoc and in a straightforward manner into an affinity resin. Its potential in omic sciences was successfully put to test in the context of the analysis of post-translational modification by S-nitrosylation with two different tissues: mature pea leaves and embryogenic sunflower hypocotyls. The identified proteins reveal an intriguing overlap among S-nitrosylation and both tyrosine nitration and thioredoxin regulation. Chloroplastidic FBPase is a paradigm of such overlap of post-translational modifications since it is reversible modified by thioredoxin and S-nitrosylation and irreversibly by tyrosine nitration. Our results suggest a complex interrelation among different modulation mechanisms mediated by NO-derived molecules.Financial Support was provided by Dirección General de Investigacion Cientıfica y Técnica (DGICYT) (CTQ2008-01754), Junta de Andalucía (P07-FQM-02899), Universidad de Jaén campus de Excelencia Internacional Agroalimentario ceiA3 and by ERDF-cofinanced grants from Ministry of Science and Innovation (BIO2012-33904) and Junta de Andalucía (research groups BIO286 and BIO192). We also acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI)