CP, T and CPT versus temporal asymmetries for entangled states of the B-d-system

The observables used in the K-system to characterize T and CPT violation are no longer useful for the Bd-system, since the width difference between the physical states is vanishingly small. We show that only Im(epsilon) and Re(delta) can survive if Delta Gamma=0, and build alternative CP-odd, CPT-odd, T-odd and temporal asymmetries for the (B_CP -> B0, B0bar) transitions. These quantities enable us to test T and CPT invariances of the effective Hamiltonian for the B-system. The method needs the CP eigenstates B_CP, which can be tagged unambiguously to order lambda^3 from the entangled states of a B-factory

Fluor-thiol Photocoupling Reaction for Developing High Performance Nucleic Acid (NA) Microarrays

Spatially controlled anchoring of NA probes onto microscope glass slides by a novel fluor-thiol coupling reaction is performed. By this UV-initiated reaction, covalent immobilization in very short times (30 s at 254 nm) is achieved with probe densities of up to 39.6 pmol/cm2. Modulating the surface hydrophobicity by combining a hydrophobic silane and a hydrophilic silane allows the fabrication of tuned surfaces where the analyte approaches only the anchored probe, which notably reduces nonspecific adsorption and the background. The generated substrates have proven clear advantages for discriminating single-base-pair mismatches, and for detecting bacterial PCR products. The hybridization sensitivity achieved by these high-performance surfaces is about 1.7 pM. Finally, this anchoring reaction is demonstrated using two additional surfaces: polytetrafluoroethylene (PTFE) and polyvinylidene fluoride (PVDF) membranes. This provides a very interesting pathway for anchoring thiolated biomolecules onto surfaces with C–F motifs via a quick clean UV reaction.Financial support from INTERBOINTER (project CTQ2013-45875-R) and BIHOLOG (Project CTQ2016-75749-R), FEDER, and GVA PROMETEO II 2014/040 is acknowledged. P.J.-M. acknowledges the Spanish Ministry of Economy, Industry and Competitiveness for the public FPI grant (Project CTQ2013-45875-R) and the cofinancing by the European Social Fund

Literature and History through a transmedia universe: didactic possibilities of the Ministry of Time

Como profesores del Grado de Educación somos conscientes de que la alfabetización transmedia forma parte de nuestra labor docente orientada a la formación de futuros profesionales que desempeñarán su labor en un contexto comunicativo mediático. El propósito de esta comunicación es presentar una herramienta específica para el transmedia en el medio docente que se inserta dentro de un Proyecto de la Universidad de Alicante: "Propuestas transmedia y gamificación aplicadas al EEES: nuevas metodología activas para implementar las competencias en Ciencias Sociales y en Literatura" y del Proyecto de Conselleria. Consideraramos las TIC como un espacio decisivo para la difusión de la docencia a través de herramientas virtuales, desarrollando propuestas didácticas en las que se implemente a través de los transmedia y la gamificación la adquisición de nuevos contenidos en dichas áreas de conocimiento. Por ello, presentamos una experiencia educativa en la que nuestros estudiantes del Máster en Profesorado de Educación Secundaria, como futuros profesionales docentes, sean capaces de incorporar en sus clases proyectos de innovación docente con intervenciones en el aula que se sustentan en los beneficios de los universos transmedia.As teachers of the Degree in Education, we are aware that transmedia literacy is part of our teaching work aimed at training of future professionals who will carry out their work in a media communicative context. The purpose of this communication is to present a specific tool for transmedia in the teaching environment that is inserted within a Project of the University of Alicante: "Transmedia and gamification proposals applied to the EEES: new active methodologies to implement the competences in Social Sciences and in Literature "and the Conselleria Project. We consider ICT as a decisive space for the dissemination of teaching through virtual tools, developing didactic proposals in which the acquisition of new contents in these areas of knowledge is implemented through transmedia and gamification. Therefore, we present an educational experience in which our students of the Master's Degree in Secondary Education, as future teaching professionals, are able to incorporate teaching innovation projects into their clases with classroom interventions based on the benefits of the universes Transmedia

Photo-click chemistry to create nucleic acids dextran-based microarrays

The final publication is available at link.springer.com[EN] In the literature, there are reports of the utilization of various hydrogels to create generic platforms for protein microarray applications. Here, a novel strategy was developed to obtain high-performance microarrays. In it, a dextran hydrogel is used to covalently immobilize oligonucleotides and proteins. This method employs aqueous solutions of dextran methacrylate (Dx-MA), which is a biocompatible photopolymerizable monomer. Capture probes are immobilized inside the hydrogel via a light-induced thiol-acrylate coupling reaction at the same time as the dextran polymer is formed. Hydrogel microarrays based on this technique were prepared on different surfaces, such as a Blu-ray Disk and polycarbonate or alkene-functionalized glass slides, and these systems showed high probe-loading capabilities and good biorecognition yields. This methodology presents advantages such as a low cost, a short analysis time, a low limit of detection, and multiplexing capabilities, among others. Confocal fluorescence microscopy analysis demonstrated that in these hydrogel-based microarrays, receptor immobilization and the biorecognition event occurred within the hydrogel and not merely on the surface.Funding from MINECO through the project BIHOLOG CTQ/2016/75749-R is acknowledged.Díaz-Betancor, Z.; Bañuls Polo, M.; Maquieira Catala, A. (2019). Photo-click chemistry to create nucleic acids dextran-based microarrays. Analytical and Bioanalytical Chemistry. 411(25):6745-6754. https://doi.org/10.1007/s00216-019-02050-3S6745675441125Heller MJ. DNA microarray technology: devices, systems, and applications. Annu Rev Biomed Eng. 2002;4:129–53. https://doi.org/10.1146/annurev.bioeng.4.020702.153438 .Sassolas A, Leca-Bouvier BD, Blum LJ. DNA biosensors and microarrays. Chem Rev. 2008;108:109–39. https://doi.org/10.1021/cr0684467 .Uttamchandani M, Neo JL, Ong BNZ, Moochhala S. Applications of microarrays in pathogen detection and biodefence. Trends Biotechnol. 2009;27:53–61. https://doi.org/10.1016/J.TIBTECH.2008.09.004 .Yu X, Schneiderhan-Marra N, Joos TO. Protein microarrays for personalized medicine. Clin Chem. 2010;56:376–87. https://doi.org/10.1373/clinchem.2009.137158 .North SH, Taitt CR. Immobilization of biomolecular probes for arrays and assay: critical aspects of biointerfaces. In: Chemoselective and bioorthogonal ligation reactions. Weinheim: Wiley-VCH; 2017. p. 459–95.Nimse S, Song K, Sonawane M, Sayyed D, Kim T. Immobilization techniques for microarray: challenges and applications. Sensors. 2014;14:22208–29. https://doi.org/10.3390/s141222208 .Cardenas-Benitez B, Djordjevic I, Hosseini S, Madou MJ, Martinez-Chapa SO. Review: Covalent functionalization of carbon nanomaterials for biosensor applications: an update. J Electrochem Soc. 2018;165:B103–17. https://doi.org/10.1149/2.0381803jes .Qu Z, Xu H, Gu H. Synthesis and biomedical applications of poly((meth)acrylic acid) brushes. ACS Appl Mater Interfaces. 2015;7:14537–51. https://doi.org/10.1021/acsami.5b02912 .Oh SJ, Hong BJ, Choi KY, Park JW. Surface modification for DNA and protein microarrays. OMICS. 2006;10:327–43. https://doi.org/10.1089/omi.2006.10.327 .Luderer F, Walschus U. Immobilization of oligonucleotides for biochemical sensing by self-assembled monolayers: thiol–organic bonding on gold and silanization on silica surfaces. In: Immobilisation of DNA on chips I. Berlin: Springer; 2005. p. 37–56.Caminade A-M. Dendrimers as biological sensors. In: Dendrimers. Chichester: Wiley; 2011. p. 375–92.Kiat NJ, Simeon F, Phon TH, Ajikumar PK. DNA-directed assembly microarray for protein and small molecule inhibitor screening. Totowa, NJ: Humana; 2011. p. 127–40.Basinska T. Reactions leading to controlled hydrophilicity/hydrophobicity of surfaces. Curr Org Chem. 2017;21(24):2479–501. https://doi.org/10.2174/1385272821666170428123013 .Weinrich D, Köhn M, Jonkheijm P, Westerlind U, Dehmelt L, Engelkamp H, et al. Preparation of biomolecule microstructures and microarrays by thiol-ene photoimmobilization. ChemBioChem. 2010;11:235–47. https://doi.org/10.1002/cbic.200900559 .Wendeln C, Rinnen S, Schulz C, Kaufmann T, Arlinghaus HF, Ravoo BJ. Rapid preparation of multifunctional surfaces for orthogonal ligation by microcontact chemistry. Chem Eur J. 2012;18:5880–8. https://doi.org/10.1002/chem.201103422 .Makaraviciute A, Ramanaviciene A. Site-directed antibody immobilization techniques for immunosensors. Biosens Bioelectron. 2013;50:460–71. https://doi.org/10.1016/j.bios.2013.06.060 .Bañuls M-J, Jiménez-Meneses P, Meyer A, Vasseur J-J, Morvan F, Escorihuela J, et al. Improved performance of DNA microarray multiplex hybridization using probes anchored at several points by thiol–ene or thiol–yne coupling chemistry. Bioconjug Chem. 2017;28:496–506. https://doi.org/10.1021/acs.bioconjchem.6b00624 .Neumann K, Conde-González A, Owens M, Venturato A, Zhang Y, Geng J, et al. An approach to the high-throughput fabrication of glycopolymer microarrays through thiol–ene chemistry. Macromolecules. 2017;50:6026–31. https://doi.org/10.1021/acs.macromol.7b00952 .Gupta N, Lin BF, Campos LM, Dimitriou MD, Hikita ST, Treat ND, et al. A versatile approach to high-throughput microarrays using thiol-ene chemistry. Nat Chem. 2010;2:138–45. https://doi.org/10.1038/nchem.478 .Rubina AY, Dementieva EI, Stomakhin AA, Darii EL, Pan’kov SV, Barsky VE, et al. Hydrogel-based protein microchips: manufacturing, properties, and applications. Biotechniques. 2003;34:1008–22. https://doi.org/10.2144/03345rr01 .Varshosaz J. Dextran conjugates in drug delivery. Expert Opin Drug Deliv. 2012;9:509–23. https://doi.org/10.1517/17425247.2012.673580 .Desmet C, Blum LJ, Marquette CA. High-throughput multiplexed competitive immunoassay for pollutants sensing in water. Anal Chem. 2012;84:10267–76. https://doi.org/10.1021/ac302133u .Moschallski M, Evers A, Brandstetter T, Rühe J. Sensitivity of microarray based immunoassays using surface-attached hydrogels. Anal Chim Acta. 2013;781:72–9. https://doi.org/10.1016/j.aca.2013.04.013 .Beyer A, Pollok S, Berg A, Weber K, Popp J. Easy daylight fabricated hydrogel Array for colorimetric DNA analysis. Macromol Biosci. 2014;14:889–98. https://doi.org/10.1002/mabi.201300487 .Alonso R, Jiménez-Meneses P, García-Rupérez J, Bañuls M-J, Maquieira Á. Thiol–ene click chemistry towards easy microarraying of half-antibodies. Chem Commun. 2018;54:6144–7. https://doi.org/10.1039/C8CC01369A .Escorihuela J, Bañuls MJ, Grijalvo S, Eritja R, Puchades R, Maquieira Á. Direct covalent attachment of DNA microarrays by rapid thiol-ene “click” chemistry. Bioconjug Chem. 2014;25:618–27.Escorihuela J, Bañuls M-J, Puchades R, Maquieira Á. Site-specific immobilization of DNA on silicon surfaces by using the thiol–yne reaction. J Mater Chem B. 2014;2:8510–7. https://doi.org/10.1039/C4TB01108B .Escorihuela J, Bañuls MJ, Puchades R, Maquieira Á. Development of oligonucleotide microarrays onto Si-based surfaces via thioether linkage mediated by UV irradiation. Bioconjug Chem. 2012;23:2121–8.Dondoni A. The emergence of thiol–ene coupling as a click process for materials and bioorganic chemistry. Angew Chem Int Ed. 2008;47:8995–7. https://doi.org/10.1002/anie.200802516 .Mira D, Llorente R, Morais S, Puchades R, Maquieira A, Marti J. High-throughput screening of surface-enhanced fluorescence on industrial standard digital recording media. Proc SPIE. 2004;5617:364–73.van Dijk-Wolthuis WNE, Franssen O, Talsma H, van Steenbergen MJ, Kettenes-van den Bosch JJ, Hennink WE. Synthesis, characterization, and polymerization of glycidyl methacrylate derivatized dextran. Macromolecules. 1995;28:6317–22. https://doi.org/10.1021/ma00122a044 .Pirrung MC. How to make a DNA chip. Angew Chem Int Ed. 2002;41:1276–89. https://doi.org/10.1002/1521-3773(20020415)41:83.0.CO;2-2 .Wang C, Jia X-M, Jiang C, Zhuang G-N, Yan Q, Xiao S-J. DNA microarray fabricated on poly(acrylic acid) brushes-coated porous silicon by in situ rolling circle amplification. Analyst. 2012;137:4539. https://doi.org/10.1039/c2an35417a .Casanova-Salas I, Rubio-Briones J, Calatrava A, Mancarella C, Masiá E, Casanova J, et al. Identification of miR-187 and miR-182 as biomarkers of early diagnosis and prognosis in patients with prostate cancer treated with radical prostatectomy. J Urol. 2014;192:252–9. https://doi.org/10.1016/j.juro.2014.01.107

Holographic Recording of Unslanted Volume Transmission Gratings in Acrylamide/Propargyl Acrylate Hydrogel Layers: Towards Nucleic Acids Biosensing

[EN] The role of volume hydrogel holographic gratings as optical transducers in sensor devices for point-of-care applications is increasing due to their ability to be functionalized for achieving enhanced selectivity. The first step in the development of these transducers is the optimization of the holographic recording process. The optimization aims at achieving gratings with reproducible diffraction efficiency, which remains stable after reiterative washings, typically required when working with analytes of a biological nature or several step tests. The recording process of volume phase transmission gratings within Acrylamide/Propargyl Acrylate hydrogel layers reported in this work was successfully performed, and the obtained diffraction gratings were optically characterized. Unslanted volume transmission gratings were recorded in the hydrogel layers diffraction efficiencies; up to 80% were achieved. Additionally, the recorded gratings demonstrated stability in water after multiple washing steps. The hydrogels, after functionalization with oligonucleotide probes, yields a specific hybridization response, recognizing the complementary strand as demonstrated by fluorescence. Analyte-sensitive hydrogel layers with holographic structures are a promising candidate for the next generation of in vitro diagnostic tests.This work was financially supported by the E.U. FEDER, the Spanish Ministry of Science and Innovation (ADBIHOL-PID2019-110713RB-I00/AEI/10.13039/501100011033) and Generalitat Valenciana (PROMETEO/2020/094). M. I. Lucio acknowledges MINECO for her Juan de la Cierva-Incorporacion grants (IJC 2018-035355-I). P. Zezza acknowledges the Generalitat Valenciana for her S. Grisolia grant and the UPV for the mobility grant (BEFPI 2022). Funding was also received from Aid for First Research Projects (PAID-06-22) and the Vice-rectorate for Research of the Universitat Politecnica de Valencia (UPV). Partially funded by the European Space Agency, through PEA4000129503 collaborative project: Wound Healing In Space: Key challenges towards Intelligent and Enabling Sensing platforms.Zezza, P.; Lucío, MI.; Naydenova, I.; Bañuls Polo, M.; Maquieira Catala, A. (2023). Holographic Recording of Unslanted Volume Transmission Gratings in Acrylamide/Propargyl Acrylate Hydrogel Layers: Towards Nucleic Acids Biosensing. Gels. 9(9). https://doi.org/10.3390/gels90907109

Teacher Motivation: Exploring the Integration of Technology and Didactics in the Narratives of Future Teachers

Different theories addressing the motivational process in educational practice highlight the importance of the teacher’s perspective in the effective integration of technologies as pedagogical–didactic tools in the classroom. The current study consists of a manifest content analysis applying a non-experimental, cross-sectional, qualitative research design. A longitudinal study was conducted through semi-structured individual interviews over three academic years (2020–2023) with a sample of 36 teacher-training students, including undergraduate and postgraduate students from the Valencian Community. The results obtained reveal the participants’ perceptions of the motivations for using technology in their self-learning process, highlighting the positive value they attribute to these tools as resources to motivate their future students. The narratives compiled highlight the relationship between the use of technology and improved academic performance in teacher training. Furthermore, they underline the need to incorporate educational models that not only foster digital skills but also provide solid pedagogical training in this area.This research was funded by the Conselleria de Innovación, Universidades, Ciencia y Sociedad Digital de la Generalitat Valencia to support and promote the activity of emerging I+D+I: La brecha digital de género y el modelo TPACK en la formación del profesorado: análisis de la capacitación digital docente (reference GV/2021/077)

Surface Micro-Patterned Biofunctionalized Hydrogel for Direct Nucleic Acid Hybridization Detection

[EN] The present research is focused on the development of a biofunctionalized hydrogel with a surface diffractive micropattern as a label-free biosensing platform. The biosensors described in this paper were fabricated with a holographic recording of polyethylene terephthalate (PET) surface micro-structures, which were then transferred into a hydrogel material. Acrylamide-based hydrogels were obtained with free radical polymerization, and propargyl acrylate was added as a comonomer, which allowed for covalent immobilization of thiolated oligonucleotide probes into the hydrogel network, via thiol-yne photoclick chemistry. The comonomer was shown to significantly contribute to the immobilization of the probes based on fluorescence imaging. Two different immobilization approaches were demonstrated: during or after hydrogel synthesis. The second approach showed better loading capacity of the bioreceptor groups. Diffraction efficiency measurements of hydrogel gratings at 532 nm showed a selective response reaching a limit of detection in the complementary DNA strand of 2.47 mu M. The label-free biosensor as designed could significantly contribute to direct and accurate analysis in medical diagnosis as it is cheap, easy to fabricate, and works without the need for further reagents.This work was financially supported by the E.U. FEDER, the Spanish Ministry of Science and Innovation (ADBIHOL-PID2019-110713RB-I00/AEI/10.13039/501100011033) and Generalitat Valenciana (PROMETEO/2020/094). M. I. Lucío acknowledges her Juan de la Cierva-Incorporación grant (IJC 2018-035355-I) funded by MCIN/AEI/10.13039/501100011033. P. Zezza acknowledges Generalitat Valenciana for her Grisolia fellowship grant.Zezza, P.; Lucío, MI.; Fernández, E.; Maquieira Catala, A.; Bañuls Polo, M. (2023). Surface Micro-Patterned Biofunctionalized Hydrogel for Direct Nucleic Acid Hybridization Detection. Biosensors. 13(3). https://doi.org/10.3390/bios1303031213

Novel and rapid activation of polyvinylidene fluoride membranes by UV light

[EN] Polyvinylidene fluoride (PVDF) membranes have become essential because of their huge applicability to the industry; however, they still present some limitations. This study focuses on the modification of PVDF membrane properties such as hydrophobicity, wettability, and functionality. To obtain a stable grafting, the surface of the membrane is hydroxylated using UV light at 254¿nm, followed by covalent immobilization of (3-aminopropyl)triethoxysilane (APTES) and vinyltriethoxysilane (VTES). The physicochemical and morphological properties of modified and raw PVDF membranes were analyzed by spectroscopy, microscopy, and goniometry. Finally, nucleic acid microarray technology results showed that PVDF and PVDF-VTES membranes had probe immobilization densities of 5 and 11¿pmol/cm2 and hybridization limits of detection of 1 and 5¿nM, respectively.Financial support from Spanish Ministry of Economy and Competitiveness (BIHOLOG Project CTQ2016-75749-R) and FEDER is acknowledged. P.J.-M. acknowledges the Spanish Ministry of Economy, Industry and Competitiveness for the public FPI grant (Project CTQ2013-45875-R) and cofinancing by the European Social Fund.Jimenez-Meneses, P.; Bañuls Polo, M.; Puchades, R.; Maquieira Catala, A. (2019). Novel and rapid activation of polyvinylidene fluoride membranes by UV light. Reactive and Functional Polymers. 140:56-61. https://doi.org/10.1016/j.reactfunctpolym.2019.04.012S566114

Fluor-thiol Photocoupling Reaction for Developing High Performance Nucleic Acid (NA) Microarrays

[EN] Spatially controlled anchoring of NA probes onto microscope glass slides by a novel fluor-thiol coupling reaction is performed. By this UV-initiated reaction, covalent immobilization in very short times (30 s at 254 nm) is achieved with probe densities of up to 39.6 pmol/cm(2). Modulating the surface hydrophobicity by combining a hydrophobic silane and a hydrophilic silane allows the fabrication of tuned surfaces where the analyte approaches only the anchored probe, which notably reduces nonspecific adsorption and the background. The generated substrates have proven clear advantages for discriminating single-base-pair mismatches, and for detecting bacterial PCR products. The hybridization sensitivity achieved by these high-performance surfaces is about 1.7 pM. Finally, this anchoring reaction is demonstrated using two additional surfaces: polytetrafluoroethylene (PTFE) and polyvinylidene fluoride (PVDF) membranes. This provides a very interesting pathway for anchoring thiolated biomolecules onto surfaces with C-F motifs via a quick clean UV reaction.Financial support from INTERBOINTER (project CTQ2013-45875-R) and BIHOLOG (Project CTQ2016-75749-R), FEDER, and GVA PROMETEO II 2014/040 is acknowledged. The authors also thank Dr. Tortajada-Genaro and Dr. Ninoles Rodenes for providing the Salmonella and Campylobacter PCR-amplified products. P.J.-M. acknowledges the Spanish Ministry of Economy, Industry and Competitiveness for the public FPI grant (Project CTQ2013-45875-R) and the cofinancing by, the European Social Fund. Dr. Miguel Angel Gonzalez-Martinez, Dr. Sergio Navalon, and Dr. Patricia Concepcion from Universitat Politecnica de Valencia are acknowledged for their help in the XPS analysis.Jimenez-Meneses, P.; Bañuls Polo, M.; Puchades, R.; Maquieira Catala, A. (2018). Fluor-thiol Photocoupling Reaction for Developing High Performance Nucleic Acid (NA) Microarrays. Analytical Chemistry. 90(19):11224-11231. https://doi.org/10.1021/acs.analchem.8b00265S1122411231901

Site-specific immobilization of DNA on silicon surfaces by using the thiol-yne reaction

[EN] Covalent immobilization of ssDNA fragments onto silicon-based materials was performed using the thiol-yne reaction. Chemical functionalization provided alkyne groups on the surface where the thiol-modified oligonucleotide probes can be easily photoattached as microarrays, reaching an immobilization density around 30 pmol cm(-2). The developed method presents the advantages of spatially controlled probe anchoring (by using a photomask), direct attachment without using cross-linkers, and short irradiation times (20 min). Hybridization efficiencies up to 70%, with full complementary strands, were reached. The approach was evaluated by scoring single nucleotide polymorphisms with a discrimination ratio around 15. Moreover, the potential applicability of the proposed methodology is demonstrated through the specific detection of 20 nM of a genomic target of bacterial Escherichia coli.This research was supported by Ministerio de Ciencia e Innovacion (CTQ2013-45875-R) and Generalitat Valenciana (PROMETEO/2010/008).Escorihuela Fuentes, J.; Bañuls Polo, M.; Puchades, R.; Maquieira Catala, Á. (2014). Site-specific immobilization of DNA on silicon surfaces by using the thiol-yne reaction. Journal of Materials Chemistry B. 2(48):8510-8517. https://doi.org/10.1039/c4tb01108bS8510851724