https://hdl.handle.net/10481/63754 2026-04-14T14:02:08Z https://hdl.handle.net/10481/103813 Electrochemical immunoplatform for the quantification of epithelial extracellular vesicles applied to prostate cancer diagnosis Felici, Emiliano; González Martínez, Coral; Valero Griñán, María Teresa; Gato-Zambrano, Sheila; Pereira, Sirley; Fernández-Baldo, Martín-A.; Ortega Sánchez, Francisco Gabriel Prostate cancer (PCa) is the second most commonly diagnosed cancer in men worldwide, and its early detection is critical for improving patient outcomes through timely and effective treatment. In this work, we present the first electrochemical immunoplatform based on magnetic microbeads (MBs) for the determination of epithelial extracellular vesicles (EpEVs), which are emerging as promising biomarkers for PCa diagnosis and prognosis. The immunoplatform employs MBs functionalized with anti-EpCAM antibodies to selectively capture EpEVs, forming sandwich-type immune complexes that are detected via amperometry at disposable screen-printed carbon electrodes. The method demonstrated a detection limit of 0.4 ng µL⁻¹ of EpEVs obtained from PC-3 cell line´s culture, excellent reproducibility (coefficient of variation < 5%), and high selectivity against potential interferences. Comparative analysis with colorimetric immune-magnet ELISA test showed a strong correlation between the two methods, confirming the reliability of the proposed approach. Furthermore, the electrochemical platform provided better precision and a lower limit of detection than the immune magnet ELISA method, indicating its superior analytical performance. Clinical validation using patient samples revealed that the combination of EpEV detection with PSA levels significantly improves the sensitivity and specificity of PCa diagnosis. This novel immunoplatform represents a promising analytical tool for early detection and monitoring of PCa, with potential applications in personalized cancer management. Universidad Nacional de San Luis (PROICO 02–2220), Agencia Nacional de Promoción Científica y Tecnológica (PICT-2021-GRF-TI-00136). Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET) (PIP–11220200100033CO). Instituto de Salud Carlos III (ISCIII) (PI22_01275, CP23/00134, DTS23_00030). Universidad de Granada (P32/22/02). https://hdl.handle.net/10481/101649 Tracking cell proliferation using a nanotechnology based approach Altea-Manzano, Patricia; Unciti-Broceta, Juan Diego; Cano Cortes, María Victoria; Ruiz-Blas, María Paz; Valero-Griñan, Mª Teresa; Díaz-Mochón, Juan José; Sánchez-Martín, Rosario María Aim: To develop an efficient nanotechnology fluorescence based method to track cell proliferation to avoid the limitations of current cell-labelling dyes. Material & methods: Synthesis, PEGylation, bifunctionalization and labelling with a fluorophore (Cy5) of 200 nm polystyrene nanoparticles (NPs) were performed. These NPs were characterised and assessed for in vitro long-term monitoring of cell proliferation. Results: The optimisation and validation of this method to track long term cell proliferation assays have been achieved with high reproducibility, without cell cycle disruption. This method has been successfully applied in several adherent and suspension cells including hard-to-transfect cells and isolated human primary lymphocytes. Conclusion: A novel approach to track efficiently cellular proliferation by flow cytometry using fluorescence labelled nanoparticles has been successfully developed. https://hdl.handle.net/10481/101645 Novel bead-based platform for direct detection of unlabelled nucleic acids through Single Nucleobase Labelling Venkateswaran, Seshasailam; Luque-González, Maria Angélica; Tabraue-Chávez, Mavys; Fara, Mario Antonio; López-Longarela, Barbara; Cano-Cortes, Victoria; López-Delgado, Francisco Javier; Sánchez-Martín, Rosario María; Ilyine, Hugh; Bradley, Mark; Pernagallo, Salvatore; Díaz-Mochón, Juan José Highlights • Direct detection of native microRNAs via Single Nucleotide Labelling. • Detection of native nucleic acids with single base resolution. • One hour assay for direct detection of miRNA with a LoD of 15 fmole. https://hdl.handle.net/10481/101570 Pyrazolopyrimide library screening in glioma cells discovers highly potent antiproliferative leads that target the PI3K/mTOR pathway Baillache, DJ; Fraser, C; Myers, SH; Unciti-Broceta, A; Valero Griñán, María Teresa The search for novel targeted inhibitors active on glioblastoma multiforme is crucial to develop new treatments for this unmet clinical need. Herein, we report the results from a screening campaign against glioma cell lines using a proprietary library of 100 structurally-related pyrazolopyrimidines. Data analysis identified a family of compounds featuring a 2-amino-1,3-benzoxazole moiety (eCF309 to eCF334) for their antiproliferative properties in the nM range. These results were validated in patient-derived glioma cells. Available kinase inhibition profile pointed to blockade of the PI3K/mTOR pathway as being responsible for the potent activity of the hits. Combination studies demonstrated synergistic activity by inhibiting both PI3Ks and mTOR with selective inhibitors. Based on the structure activity relationships identified in this study, five new derivatives were synthesized and tested, which exhibited potent activity against glioma cells but not superior to the dual PI3K/mTOR inhibitor and lead compound of the screening eCF324. https://hdl.handle.net/10481/101550 Small Molecule Kinase Inhibitor Drugs (1995–2021): Medical Indication, Pharmacology, and Synthesis Ayala, C; Valero, T; Lorente, A; Baillache, DJ; Crocke, S; Unciti-Broceta, A The central role of dysregulated kinase activity inthe etiology of progressive disorders, including cancer, has fosteredincremental efforts on drug discovery programs over the past 40years. As a result, kinase inhibitors are today one of the mostimportant classes of drugs. The FDA approved 73 small moleculekinase inhibitor drugs until September 2021, and additionalinhibitors were approved by other regulatory agencies during thattime. To complement the published literature on clinical kinaseinhibitors, we have prepared a review that recaps this large data setinto an accessible format for the medicinal chemistry community.Along with the therapeutic and pharmacological properties of eachkinase inhibitor approved across the world until 2020, we provide the synthesis routes originally used during the discovery phase,many of which were only available in patent applications. In the last section, we also provide an update on kinase inhibitor drugsapproved in 2021.