https://hdl.handle.net/10259/11210 2026-06-25T14:06:23Z https://hdl.handle.net/10259/11876 How reliable is the evaluation of DNA binding constants? Insights and best practices based on an inter-laboratory fluorescence titration study Dömötör, Orsolya; Binacchi, Francesca; Ribeiro, Nádia; Busto Vázquez, Natalia; González García, Jorge .; García España, Enrique; Correia, Isabel; Enyedy, Éva A.; Hamacek, Josef; Terenzi, Alessio; Basílio, Nuno; Barone, Giampaolo; Cavaco, Isabel; Biver, Tarita In all experimental sciences, the precision and reliability of quantitative measurements are paramount. This is particularly true when examining the interactions between small molecules and biomolecules/polyelectrolytes, such as DNAs/RNAs, and yet it is overlooked in most publications of thermodynamic binding parameters. This paper presents findings from COST Action 18202 “Network for Equilibria and Chemical Thermodynamics Advanced Research,” which assessed the consistency of data derived from the interactions of calf-thymus DNA (CT-DNA) with the fluorescent intercalator ethidium bromide (EB) through spectrofluorimetric titrations. We first discuss critical experimental aspects and propose a reference experimental protocol which can be used to calibrate procedures for the determination of nucleic acid binding equilibrium constants. We then fit the experimental points according to different procedures and analyse the results focusing on the statistical dispersion of the data, aiming at enlightening the strong and weak points of different fitting procedures. The implications of this work are significant, demonstrating how the statistical dispersion of experimental data can influence the interpretation of biochemical coordination mechanisms. Our study reveals that, despite rigorous protocol standardization, the determination of binding parameters remains sensitive to the choice of data fitting method, with deviations in the logarithmic stability constant (logK) values not falling below 5 % relative standard deviation (RSD), or ± 0.5 logK units for 95 % confidence. This variability evidences the critical need for standardized best practices in data treatment as well as experimental procedures. Although our study focuses on the EB/CT-DNA system through fluorescence titrations, the broader implications for other methodologies across various biochemical systems highlight the importance of this first-of-its-kind inter-laboratory comparison in advancing our understanding of biochemical coordination processes. 2025-02-01T00:00:00Z https://hdl.handle.net/10259/11621 Creatine plus β-Hydroxy-β-Methylbutyrate supplementation is associated with preserved glutathione redox-balance and redox–function associations in older adults: a secondary analysis of a randomized crossover trial Ramos-Hernández, Rafael; Mielgo Ayuso, Juan; Fernández-Lázaro, Diego; Abia, Alba; Pérez-López, Juan F.; Saiz Rodríguez, Miriam; Busto Vázquez, Natalia Oxidative stress contributes to age-related musculoskeletal decline, partly through disruption of glutathione-dependent redox homeostasis. Although creatine and β-hydroxy-β-methylbutyrate (HMB) have been individually linked to antioxidant and cytoprotective effects, their combined influence on systemic redox balance in older adults remains insufficiently characterized.To examine the effects of creatine plus HMB supplementation on oxidative stress biomarkers and composite redox indices, and to explore whether redox adaptations co-vary with changes in functional measures in physically active older adults.In a randomized, double-blind, placebo-controlled crossover trial, 30 physically active older adults (62.7 ± 5.3 years; 20 men, 10 women) completed two 6-week intervention phases (3 g/day creatine + 3 g/day calcium HMB vs. placebo) during supervised exercise training. Primary endpoints were oxidized glutathione and the Glutathione Redox Index. Secondary biomarkers and composite indices were analyzed with false discovery rate (FDR) control. Percent changes (Δ%) in functional tests were examined exclusively as exploratory correlates of redox adaptations.Supplementation was associated with attenuation of the placebo-related increase in oxidized glutathione and nominal preservation of the Glutathione Redox Index, although these effects did not remain significant after FDR adjustment. In men, a nominal increase in malondialdehyde was observed under supplementation. Exploratory analyses indicated weak associations between changes in composite redox indices and Δ% functional measures.Creatine plus HMB supplementation was associated with nominal modulation of glutathione-centered redox balance during training in active older adults. Exploratory redox–function associations support further investigation in larger, adequately powered trials. 2026-02-01T00:00:00Z https://hdl.handle.net/10259/11620 An Overview of Endometriosis and Potential Pharmacogenetic Targets Pérez Gómez, Noelia; Martínez-Zamora, María Ángeles; Carmona, Francisco; Novalbos, Jesús; Abad-Santos, Francisco; Koller, Dora; Saiz Rodríguez, Miriam Endometriosis is a chronic inflammatory disease characterized by the presence of endometrial-like tissue outside the uterine cavity. It affects approximately 10–15% of women of reproductive age globally and is characterized by heterogeneous symptoms with chronic pelvic pain, dysmenorrhea, and infertility being the most common. Although pharmacological treatments are available to manage its symptoms, many women either do not respond to these therapies or experience adverse drug reactions (ADRs) that outweigh the original symptoms of endometriosis. Current research highlights a critical gap in pharmacogenetic biomarkers for endometriosis treatment, limiting the potential for personalized therapeutic strategies. Integrative multi-omics approaches combining genetic, inflammatory, and hormonal profiles may enhance patient stratification and optimize individualized care. 2026-03-01T00:00:00Z https://hdl.handle.net/10259/11612 Combined creatine and β-hydroxy-β-methylbutyrate supplementation with integral conditioning exercise enhances functional performance and metabolic health in physically active older adults: A randomized controlled crossover trial Ramos-Hernández, Rafael; Busto Vázquez, Natalia; Miguel Ortega, Alvaro; Martínez Ferrán, María; Santamaría Peláez, Mirian; Saiz Rodríguez, Miriam; Mielgo Ayuso, Juan Objective: to investigate the effects of six weeks of CRE + HMB supplementation integrated with a supervised multicomponent exercise program on functional performance, metabolic efficiency, and physiological health in older adults. Conclusions: six weeks of CRE + HMB supplementation integrated with supervised multicomponent training enhanced mobility, metabolic efficiency, and selected physiological outcomes in physically active older adults. This strategy represents a safe, feasible, and practical approach to sustain functional independence and metabolic health with aging. 2026-01-01T00:00:00Z