DFIFA. Artículos del Departamento de Fisiología y Farmacología

Oxidative eustress regulates insulin signaling and promotes GLUT4-mediated glucose uptake in insulin-resistant skeletal muscle fibres

2026-06-24T00:01:31Z

[EN]The document focuses on how oxidative eustress (a moderate and beneficial level of reactive species) acts as an alternative regulatory mechanism to trigger GLUT4 trafficking and restore glucose uptake within insulin-resistant skeletal muscle.

Bioaccessibility and genoprotective effect of melanoidins obtained from common and soft bread crusts: relationship between melanoidins and their bioactivity

2026-06-20T00:01:00Z

[EN]Bread crust constitutes an important by-product of the bakery industry, and its utilization for the isolation of melanoidins to be used as a functional ingredient can enhance its added value and contribute to health. The aim of this study was to evaluate the bioaccessibility, bioactivity, and genoprotective effect of melanoidins derived from bread crust. Bioaccessibility was assessed in gastric, intestinal digestion, and colonic fermentation fractions. The results revealed a relationship between bioaccessible melanoidins and their type (common or soft bread). No cytotoxicity effects were observed for bioaccessible fractions, as assessed by MTT and RTA methods, and they did not affect the distribution of E-cadherin in Caco-2 cells, confirming their ability to maintain membrane integrity. Furthermore, our study demonstrated that the gastrointestinal and colonic fermentation fractions successfully transported across the intestinal barrier, without affecting cell permeability, and showed antioxidant activity on the basolateral side of the cell monolayer. Remarkably, both fractions displayed a significant genoprotective effect in Caco-2 cells. Our findings provide crucial insights into the relationship between the melanoidins and their bioactivity and genoprotective effect. These results demonstrated the potential of bioaccessible melanoidins as valuable bioactive compounds for the development of functional foods, without showing toxic effects on gastrointestinal cells.

Bioaccessible bread melanoidins modulate oxidative stress, reduce inflammation and suppress adhesion of helicobacter pylori to caco-2 cells

2026-06-20T00:00:56Z

[EN]Background/Objectives: Helicobacter pylori is a major contributor to gastric infections; it is prevalent in humans and associated with gastrointestinal diseases. In recent years, the increase in antimicrobial resistance has contributed to the need for alternative approaches, prompting interest in natural products with antimicrobial and antivirulence properties. This study investigated the effect of bioaccessible melanoidins from common and soft bread crust against H. pylori infection. Methods: Melanoidins were extracted using dead-end ultrafiltration, and bioaccessible fractions were obtained through in vitro digestion. The bactericidal effect of melanoidins was assessed at 2% and 4% concentrations over 24 and 48 h. The effect on H. pylori adhesion of 100 μg/mL and 200 μg/mL of gastric and intestinal bioaccessible fractions of melanoidins was evaluated in Caco-2 cells. Results: The bactericidal effect of melanoidins revealed significant efficacy, with a greater effect for soft bread melanoidins. The gastric fractions exhibited a higher inhibitory effect, which is crucial for gastric mucosa, the primary site of H. pylori infection. Both bioaccessible fractions showed anti-inflammatory and antioxidant effects against H. pylori-induced inflammation, particularly in the gastric fractions. This was evidenced by a reduction in interleukin-6 and interleukin-8 release and an enhancement in interleukin-10 release. The observed reduction in reactive oxygen species (ROS) and the maintenance of glutathione levels indicate an improved redox status. Conclusions: This study emphasizes the potential of melanoidins, especially from soft bread, as bioactive compounds against H. pylori, offering insights for future functional food development

Age and hypertension synergize with dehydration to cause renal frailty in rats and predispose them to intrinsic acute kidney injury

2026-06-20T00:00:53Z

[EN]Acute kidney frailty (AKF) is a condition of increased susceptibility to acute kidney injury (AKI), an abrupt impairment of renal excretory function potentially leading to severe complications. Prevention of AKI relies on the recognition of risk factors contributing to AKF. At the population level, dehydration constitutes a predisposing factor for AKI. However, renal frailty may be context-specific, with variations among patients in the types of damage and the distinct pathological mechanisms. In this regard, we studied the combined effect of dehydration with other factors on renal homeostasis, such as increasing age and hypertension. AKF status was studied in rats bearing risk factors individually and in combination and was evaluated as the level of AKI induced by a triggering dose of cisplatin, which is known to be mildly nephrotoxic for young, healthy rats. AKI was assessed through parameters of renal function (including creatinine, urea, creatinine clearance, proteinuria, and fractional excretion of sodium) and histopathology of renal tissue specimens. The hydration status was measured by bioelectric impedance and other techniques. Water deprivation induces a dehydration state characterized by reductions in body weight and urinary flow and increases in hematocrit and plasma and urine osmolality. Bioelectric impedance showed a net loss of body water after water deprivation with no relevant changes in body mass distribution. Dehydration is not sufficient to predispose young control rats to intrinsic AKI. However, the combination of dehydration with advanced age or hypertension induces AKF evidenced by a magnified response of renal dysfunction (reduced filtration and tubular function) and tubular necrosis caused by low-dose cisplatin treatment. This study highlights the relevance of addressing AKF as a premorbid condition providing prophylactic opportunities and shows that dehydration differentially predisposes to prerenal and intrinsic AKI.

Regulation and function of the cytosolic viral RNA sensor RIG-I in pancreatic beta cells

2026-05-20T00:02:43Z

[EN]Enteroviral infections are associated with type I diabetes. The mechanisms by which viruses or viral products such as double-stranded RNA (dsRNA) affect pancreatic beta cell function and survival remain unclear. We have shown that extracellular dsRNA induces beta cell death via Toll-like receptor-3 (TLR3) signaling whereas cytosolic dsRNA triggers the production of type I interferons and apoptosis via a TLR3-independent process. We presently examined expression of the intracellular viral RNA sensors, the RNA helicases RIG-I and MDA5, and documented the functionality of RIG-I in pancreatic beta cells. FACS-purified rat beta cells and islet cells from wild-type or TLR3(-/-) mice were cultured with or without the RIG-I-specific ligand 5'-triphosphate single-stranded RNA (5'triP-ssRNA), the synthetic dsRNA polyI:C (PIC) or 5'OH-ssRNA (negative control); the RNA compounds were added in the medium or transfected in the cells using lipofectamine. RIG-I and MDA5 expression were determined by real-time RT-PCR. NF-kappaB and IFN-beta promoter activation were studied in the presence or absence of a dominant-negative form of RIG-I (DN-RIG-I). Both extracellular (PICex) and intracellular (PICin) PIC increased expression of RIG-I and MDA5 in pancreatic beta cells. TLR3 deletion abolished PICex-induced up-regulation of the helicases in beta cells but not in dendritic cells. PICin-induced NF-kappaB and IFN-beta promoter activation were prevented by the DN-RIG-I. The RIG-I-specific ligand 5'triP-ssRNA induced IFN-beta promoter activation and beta cell apoptosis. Our results suggest that the RIG-I pathway is present and active in beta cells and could contribute to the induction of insulitis by viral RNA intermediates.

Medication review improves pain management and quality of life in chronic pain: a pilot randomized controlled study

2026-05-19T00:01:12Z

[EN]Chronic pain is a complex condition that benefits from a multidisciplinary approach. This pilot parallel-group single-blinded randomized controlled study evaluated the feasibility, acceptability and adherence by patients and physicians of pharmacist-led medication review on chronic pain patients. Trends in pain intensity, quality of life and patient satisfaction were examined. Twenty adults were recruited from two primary care units in Porto, Portugal, and randomly assigned to either the medication review (MR) group, (n = 10) using the Dader method, or the usual care (UC) group, (n = 10) and given general advice, for 16-weeks. Pain intensity decreased by 2.07 (MR group) and increased by 0.52 (UC group), yielding an adjusted mean difference of 2.77 (95% CI, -4.93 to -0.62; p = 0.008). Pain relief was reported by 62.5% in the MR group versus 37.5% in UC (p = 0.357). The MR group showed significant improvement in physical functioning (p = 0.019) and higher treatment satisfaction (p = 0.029). The acceptance rate of MR interventions was 71%, which resolved 63% of negative medication outcomes. Acceptability was high (> 90% of planned interviews). Conducting pharmacist-led MR for chronic pain management in primary care is feasible and well accepted by patients and physicians. Observed trends toward improved pain and QoL warrant confirmation in a larger trial. This pilot trial was registered in clinicaltrials.gov (NCT06997861).

Sex-dependent modulation of CGRPergic neurovascular activity by 5-CT in rats

2026-05-06T00:01:27Z

[EN]Background and purpose: Serotonin modulates vascular tone both directly and indirectly through autonomic and sensory nerves innervating blood vessels. Perivascular sensory nerves release calcitonin gene-related peptide (CGRP), a potent vasodilator strongly implicated in migraine pathophysiology. In male rats, the serotonergic system inhibits CGRPergic vasodepressor responses via 5-HT1B/1F and 5-HT7 receptors. Since both serotonergic and CGRPergic pathways exhibit marked sex differences, the present study investigated the 5-HT receptor (sub)types involved in the 5-carboxamydotryptamine (5−CT, 5-HT1/5/7 receptor agonist) modulation of vascular CGRPergic neurotransmission in rats, focusing on sex-dependent differences. Methods: Male and female Wistar rats (14–16 weeks old) were pithed and pretreated with an i.v. continuous infusion of hexamethonium and methoxamine, followed by administration of 5-HT-related drugs. Mean blood pressure (MBP) and heart rate (HR) were continuously recorded throughout the experiments. Vasodepressor CGRPergic responses were elicited by electrical stimulation of the sensory outflow (0.1–5 Hz) or i.v. α-CGRP (0.1–1 μg/kg). Results: Basal MBP and HR were lower in females than in males, whereas the methoxamine-induced increase in MBP was greater in females. The electrically evoked vasodepressor responses, as well as their inhibition by 5−CT, were similar in both sexes. In males, the inhibitory effect of 5−CT was reproduced by 5-HT1B, 5-HT1F, and 5-HT7 receptor agonists (CP-93,129, LY344864, and AS-19, respectively) and persisted in the presence of the 5-HT5A receptor antagonist SB699551. In contrast, in females, 5-CT-induced inhibition was mimicked by 5-HT1F and 5-HT7 receptor agonists and was not affected by administration of SB699551. None of the other 5-HT receptor agonists (5-HT1A/1B/1D) modified the CGRPergic vasodilator responses in females. Only AS-19 reduced the vasodepressor responses elicited by exogenous α-CGRP in females. Conclusion: 5−CT inhibits perivascular sensory CGRPergic neurotransmission in both male and female rats. Unlike males, where the 5−CT effect is mediated by prejunctional 5-HT1B/1F/7 receptors, in females, this inhibitory effect is mediated by prejunctional 5-HT1F and pre and/or postjunctional 5-HT7 receptors. These findings provide novel insights into sex-specific serotonergic modulation of neurovascular function.

Angiotensin II and EDH pathways underlie the vascular sympatho-modulation by 5-HT in female rats

2026-05-06T00:01:22Z

[EN]The vascular 5-HT sympatho-modulation may involve inhibitory or potentiating pathways: nitric oxide (NO), endothelium-dependent hyperpolarization (EDH)-K+ channels, prostanoids, angiotensin II (Ang-II), or endothelin. Compared to males, female rats show differences in the serotonergic sympatho-regulation; therefore, we aimed to study the involvement of indirect pathways via 5-HT1D-mediated inhibition and 5-HT2A/3-mediated potentiation of vascular noradrenergic neurotransmission in females. An i.v. bolus of different inhibitors/blockers of modulators/mediators (NO, K+ channels, prostanoids, Ang-II, or endothelin) was administered prior to the infusion of the agonists, L-694,247 (5-HT1D), TCB-2 (5-HT2A), or 1-PBG (5-HT3), in female pithed rats. In these conditions, the vascular sympathetic outflow was electrically stimulated to assess the vasopressor responses. The L-694,247 vascular sympatho-inhibition was abolished by a non-selective K+ channel blocker, tetraethylammonium. The 1-PBG sympatho-excitatory vascular effect was not modified by any of the inhibitors tested, whereas TCB-2 sympatho-potentiation was blocked solely by losartan (Ang-II type 1 receptor antagonist). Moreover, Ang-II levels were increased after TCB-2 infusion in females. The EDH pathway mediates the 5-HT1D-induced sympatho-inhibition, while the 5-HT2A-evoked sympatho-excitatory effect is associated with Ang-II. In contrast, the 5-HT3 sympatho-potentiation does not involve any indirect pathway. These findings advance current understanding of the complex interactions between 5-HT and vascular homeostasis in female rats.

Targeting a Glioblastoma Cancer Stem-Cell Population Defined by EGF Receptor Variant III

2026-04-17T00:01:08Z

[EN]The relationship between mutated proteins and the cancer stem-cell population is unclear. Glioblastoma tumors frequently express EGFRvIII, an EGF receptor (EGFR) variant that arises via gene rearrangement and amplification. However, expression of EGFRvIII is restricted despite the prevalence of the alteration. Here, we show that EGFRvIII is highly coexpressed with CD133 and that EGFRvIII+/CD133+ defines the population of cancer stem cells (CSC) with the highest degree of self-renewal and tumor-initiating ability. EGFRvIII+ cells are associated with other stem/progenitor markers, whereas markers of differentiation are found in EGFRvIII− cells. EGFRvIII expression is lost in standard cell culture, but its expression is maintained in tumor sphere culture, and cultured cells also retain the EGFRvIII+/CD133+ coexpression, self-renewal, and tumor initiating abilities. Elimination of the EGFRvIII+/CD133+ population using a bispecific antibody reduced tumorigenicity of implanted tumor cells better than any reagent directed against a single epitope. This work demonstrates that a mutated oncogene can have CSC-specific expression and be used to specifically target this population.

Evolutionary Origins of Metabolic Reprogramming in Cancer

2026-04-17T00:01:00Z

[EN]Metabolic changes that facilitate tumor growth are one of the hallmarks of cancer. These changes are not specific to tumors but also take place during the physiological growth of tissues. Indeed, the cellular and tissue mechanisms present in the tumor have their physiological counterpart in the repair of tissue lesions and wound healing. These molecular mechanisms have been acquired during metazoan evolution, first to eliminate the infection of the tissue injury, then to enter an effective regenerative phase. Cancer itself could be considered a phenomenon of antagonistic pleiotropy of the genes involved in effective tissue repair. Cancer and tissue repair are complex traits that share many intermediate phenotypes at the molecular, cellular, and tissue levels, and all of these are integrated within a Systems Biology structure. Complex traits are influenced by a multitude of common genes, each with a weak effect. This polygenic component of complex traits is mainly unknown and so makes up part of the missing heritability. Here, we try to integrate these different perspectives from the point of view of the metabolic changes observed in cancer.

Intermediate molecular phenotypes to identify genetic markers of anthracycline-induced cardiotoxicity risk

2026-04-15T00:01:04Z

[EN]Cardiotoxicity due to anthracyclines (CDA) affects cancer patients, but we cannot predict who may suffer from this complication. CDA is a complex trait with a polygenic component that is mainly unidentified. We propose that levels of intermediate molecular phenotypes (IMPs) in the myocardium associated with histopathological damage could explain CDA susceptibility, so variants of genes encoding these IMPs could identify patients susceptible to this complication. Thus, a genetically heterogeneous cohort of mice (n = 165) generated by backcrossing were treated with doxorubicin and docetaxel. We quantified heart fibrosis using an Ariol slide scanner and intramyocardial levels of IMPs using multiplex bead arrays and QPCR. We identified quantitative trait loci linked to IMPs (ipQTLs) and cdaQTLs via linkage analysis. In three cancer patient cohorts, CDA was quantified using echocardiography or Cardiac Magnetic Resonance. CDA behaves as a complex trait in the mouse cohort. IMP levels in the myocardium were associated with CDA. ipQTLs integrated into genetic models with cdaQTLs account for more CDA phenotypic variation than that explained by cda-QTLs alone. Allelic forms of genes encoding IMPs associated with CDA in mice, including AKT1, MAPK14, MAPK8, STAT3, CAS3, and TP53, are genetic determinants of CDA in patients. Two genetic risk scores for pediatric patients (n = 71) and women with breast cancer (n = 420) were generated using machine-learning Least Absolute Shrinkage and Selection Operator (LASSO) regression. Thus, IMPs associated with heart damage identify genetic markers of CDA risk, thereby allowing more personalized patient management.

The role of GAB1 in cancer

2026-04-15T00:01:13Z

[EN]GRB2-associated binder 1 (GAB1) is the inaugural member of the GAB/DOS family of pleckstrin homology (PH) domain-containing proteins. Upon receiving various stimuli, GAB1 transitions from the cytoplasm to the membrane where it is phosphorylated by a range of kinases. This event recruits SH2 domain-containing proteins like SHP2, PI3K's p85 subunit, CRK, and others, thereby activating distinct signaling pathways, including MAPK, PI3K/AKT, and JNK. GAB1-deficient embryos succumb in utero, presenting with developmental abnormalities in the heart, placenta, liver, skin, limb, and diaphragm myocytes. Oncogenic mutations have been identified in the context of cancer. GAB1 expression levels are disrupted in various tumors, and elevated levels in patients often portend a worse prognosis in multiple cancer types. This review focuses on GAB1's influence on cellular transformation particularly in proliferation, evasion of apoptosis, metastasis, and angiogenesis-each of these processes being a cancer hallmark. GAB1 also modulates the resistance/sensitivity to antitumor therapies, making it a promising target for future anticancer strategies.

The use of Pharmacology-Focused journal clubs in health sciences education: a descriptive study

2026-04-15T00:01:09Z

[EN]Introduction: Physicians and nurses have a responsibility to provide evidence-based care to patients, which requires continuing education. The journal club model is an educational and collaborative tool widely used in healthcare for this purpose. However, further studies investigating the effectiveness and perception of participants are needed. The aim of this study is to explore the experience of medical and nursing students with the journal club approach applied to the subject of pharmacology. Methods: Journal club sessions were developed within the pharmacology class of the medical and nursing degrees during the academic year 2023-2024. A total of 208 students participated in the study. 90 students (43.3%) were enrolled in the nursing degree course, while 118 (56.7%) were enrolled in the medical degree course. Students' perceptions were assessed through a qualitative questionnaire of 30 questions with 3 options. Results: This study validated that this format is effective as a learning model, enhancing educational competences and non-educational competences required in health professionals. Our results showed the good acceptance of this format. Conclusion: It is concluded that the journal club strategy enhances the educational and non-educational skills necessary for doctors and nurses to develop the competencies required in today's world.

Nursing-led strategy to combat antimicrobial resistance: multi-method design

2026-04-15T00:01:07Z

[EN]Background: Antimicrobial resistance is currently a global health threat. Numerous efforts have been made to prevent or mitigate this phenomenon, but all have been insufficient. Although numerous studies have postulated nursing as a potential mediator to address this problem, no research has been conducted to put it into practice. This study aims to contextualize the problem of inappropriate antibiotic use and antimicrobial resistance, as well as to develop and implement a nurse-led educational intervention aimed at mitigating it, emphasizing the role of nurses in health education. Methods: Two different groups of participants were recruited using convenience sampling. Nurses administered a survey to 782 citizens in a population from Spain to assess their knowledge of the correct use of antibiotics and antimicrobial resistance. After completing the survey, nurses explained to the participants how to use antibiotics correctly and the problem of antimicrobial resistance. Furthermore, an educational intervention led by nurses was carried out with 104 adolescents, consisting of an oral presentation to raise awareness about the issue. The effectiveness of this intervention was evaluated through a comparative analysis before and after the activity (pre-test and post-test). Results: Our results indicated that the level of knowledge about the correct use of antibiotics and antimicrobial resistance is not statistically significant related to sex in the general population (p > 0.05). However, it is statistically significant related to age (p < 0.05), educational level (p < 0.0001) and study area (p < 0.0001). In addition, the nurse-led educational intervention increased significantly the level of knowledge on the topic among adolescents (p < 0.0001). Conclusions: These findings highlight the low level of knowledge in the population about the correct use of antibiotics and antimicrobial resistance. It also demonstrates how nurses, through their role in health education, can actively contribute to addressing the issue, providing a rationale for the inclusion of nursing in the design and implementation of strategies to prevent or mitigate antimicrobial resistance.

Development of an EGFRvIII specific recombinant antibody

2026-04-11T00:01:21Z

[EN]Background: EGF receptor variant III (EGFRvIII) is the most common variant of the EGF receptor observed in human tumors. It results from the in frame deletion of exons 2-7 and the generation of a novel glycine residue at the junction of exons 1 and 8. This novel juxtaposition of amino acids within the extra-cellular domain of the EGF receptor creates a tumor specific and immunogenic epitope. EGFRvIII expression has been seen in many tumor types including glioblastoma multiforme (GBM), breast adenocarcinoma, non-small cell lung carcinoma, ovarian adenocarcinoma and prostate cancer, but has been rarely observed in normal tissue. Because this variant is tumor specific and highly immunogenic, it can be used for both a diagnostic marker as well as a target for immunotherapy. Unfortunately many of the monoclonal and polyclonal antibodies directed against EGFRvIII have cross reactivity to wild type EGFR or other non-specific proteins. Furthermore, a monoclonal antibody to EGFRvIII is not readily available to the scientific community. Results: In this study, we have developed a recombinant antibody that is specific for EGFRvIII, has little cross reactivity for the wild type receptor, and which can be easily produced. We initially designed a recombinant antibody with two anti-EGFRvIII single chain Fv’s linked together and a human IgG1 Fc component. To enhance the specificity of this antibody for EGFRvIII, we mutated tyrosine H59 of the CDRH2 domain and tyrosine H105 of the CDRH3 domain to phenylalanine for both the anti-EGFRvIII sequence inserts. This mutated recombinant antibody, called RAbDMvIII, specifically detects EGFRvIII expression in EGFRvIII expressing cell lines as well as in EGFRvIII expressing GBM primary tissue by western blot, immunohistochemistry (IHC) and immunofluorescence (IF) and FACS analysis. It does not recognize wild type EGFR in any of these assays. The affinity of this antibody for EGFRvIII peptide is 1.7 × 107 M-1 as determined by enzyme-linked immunosorbent assay (ELISA). Conclusion: This recombinant antibody thus holds great potential to be used as a research reagent and diagnostic tool in research laboratories and clinics because of its high quality, easy viability and unique versatility. This antibody is also a strong candidate to be investigated for further in vivo therapeutic studies.

Pathophysiological Integration of Metabolic Reprogramming in Breast Cancer

2026-04-11T00:01:20Z

[EN]Metabolic changes that facilitate tumor growth are one of the hallmarks of cancer. The triggers of these metabolic changes are located in the tumor parenchymal cells, where oncogenic mutations induce an imperative need to proliferate and cause tumor initiation and progression. Cancer cells undergo significant metabolic reorganization during disease progression that is tailored to their energy demands and fluctuating environmental conditions. Oxidative stress plays an essential role as a trigger under such conditions. These metabolic changes are the consequence of the interaction between tumor cells and stromal myofibroblasts. The metabolic changes in tumor cells include protein anabolism and the synthesis of cell membranes and nucleic acids, which all facilitate cell proliferation. They are linked to catabolism and autophagy in stromal myofibroblasts, causing the release of nutrients for the cells of the tumor parenchyma. Metabolic changes lead to an interstitium deficient in nutrients, such as glucose and amino acids, and acidification by lactic acid. Together with hypoxia, they produce functional changes in other cells of the tumor stroma, such as many immune subpopulations and endothelial cells, which lead to tumor growth. Thus, immune cells favor tissue growth through changes in immunosuppression. This review considers some of the metabolic changes described in breast cancer.