Determinación de Cis-/Trans-Estilbenos en cera de panal de abeja

Las abejas juegan un papel fundamental no solo en lo relacionado con la producción de la colmena sino también en la polinización de plantas. Por todo ello, lo que las afecte negativamente, tendrá sin duda una gran repercusión en la Agricultura y el Medio Ambiente. La investigación sobre el tema se centra actualmente en la búsqueda de compuestos naturales o sintéticos alternativos al uso de antibióticos prohibidos actualmente para el tratamiento de infecciones. Un ejemplo claro es el uso de fitoalexinas, más en concreto de resveratrol que ha estado presente desde hace siglos en la medicina tradicional.Química AnalíticaMáster en Técnicas Avanzadas en Químic

Study of time-frequency characteristics of single snores: extracting new information for sleep apnea diagnosis

Obs tructive sleep apnea (OSA) is a highly prevalent chronic disease, especially in elderly and obese population . Despite constitut ing a huge health and economic problem, most patients remain undiagnosed due to limitations in current strategies. Therefore, it is essential to find cost - effective diagnostic alternatives. One of these novel approaches is the analysis of acoustic snoring signals. Snoring is an early symptom of OSA which carr ies pathophysiological information of high diagnostic value. For this reason, the main objective of this work is to study the characteristics of single snores of different types, from healthy and OSA subjects. To do that, we analyzed snoring signals from p revious databases and developed an experimental protocol to record simulated OSA - related sounds and characterize the response of two commercial tracheal microphones. Automatic programs for filtering, downsampling, event detection and time - frequency analysi s were built in MATLAB. We found that time - frequency maps and spectral parameters (central, mean and peak frequency and energy in the 100 - 500 Hz band) allow distinguishing regular snores of healthy subjects from non - regular snores and snores of OSA subject s. Regarding the two commercial microphones, we f ound that one of them was a suitable snoring sensor, while the other had a too restricted frequency response. Future work shall include a higher number of episode s and subjects , but our study has contributed to show how important the differences between regular and non - regular snores can be for OSA diagnosis, and how much clinically relevant information can be extracted from time - frequency maps and spectral parameters of single snoresPeer ReviewedPostprint (published version

Análisis exploratorio de la percepción en niños preescolares sobre la interacción recíproca con sus madres

Objective. Although conceptual advances and empirical evidence with school-age children and adolescents suggest that children’s perceptions are relevant in understanding the reciprocal reactions between parents and children, the level of knowledge in the case of pre - school children is almost non - existent. The main objective of this study was to explore the perception of preschool children of the relationships with their mothers, and to determine whether scores were related with differences on parenting competence, as well as with other parenting factors. Method. Data were collected from 50 mothers who responded to several questionnaires and their children ( n = 50) , who answered a pictorial test of perception of the relationships with their mothers. Results. Results indicated that scores from the questionnaires were consistent with perceptions of children regarding appropriate or inappropriate interactions: for mothers on parental competence, stress, and subjective well - being, and for children on psychosocial adaptation, stress, and behavior problems; as well as with scores relating to the temperament of both. Conclusion. The results and their implications are discussed in terms of establishing coherence between conceptual advances and the empirical evidence on reciprocal interactions.Objetivo. Aunque los avances conceptuales y las evidencias empíricas con niños en edad escolar y adolescentes sugieren que las percepciones de los niños son relevantes para comprender las interacciones recíprocas padres - hijos, el estado de conocimiento es prácticamente nulo en niños preescolares. El principal objetivo de este estudio buscó explorar la percepción de niños preescolares sobre las relaciones con sus madres y analizar si los puntajes diferenciales de la percepción de los niños indican diferencias en la competencia parental, así como en otros factores que tienen relación con su crianza. Método . La muestra estuvo conformada por 50 mamás que contestaron los autorreportes y sus niños ( n = 50) que contestaron una prueba pictórica de percepción de las relaciones con sus madres. Resultados . Los resultados indicaron que las percepciones de los niños sobre las interacciones adecuadas o inadecuadas fueron congruentes con los puntajes de los cuestionarios, en particular, con la competencia parental, el estrés y el bienestar subjetivo maternos y con la adaptación psicosocial, el estrés y los problemas de comportamiento de los niños, así como con el temperamento de ambos. Conclusión . Se discuten los resultados y sus implicaciones para establecer congruencia entre el avance conceptual y las evidencias empíricas sobre las interacciones recíprocas.Escopo . Ainda que os avanços conceituais e as evidencias empíricas com crianças em idade escolar e adolescentes sugerem que as percepções das crianças são relevantes para compreender as interações recíprocas pais-filhos, o estado de conhecimento é praticamente nulo com crianças pré - escolares. O principal escopo de este estudo foi o de explorar a percepção de crianças pré - escolares sobre as relações com suas mães e analisar sim pontuações diferentes da percepção das crianças indicam diferencias na competência parental, assim como em outros fatores que tem relação com a educação. Metodologia . A mostra foi formada com 50 mães que contestaram os auto - reportes e suas crianças ( n = 50) que contestaram uma prova pictórica de percepção das relações com suas mães. Resultados . Os resultados indicaram que as percepções das crianças sobre as interações adequadas ou inadequadas foram congruentes com as pontuações dos questionários; em particular, com a competência parental, o stress e o bem-estar subjetivo maternos e com a adaptação psicossocial, o stress e os problemas de comportamento das crianças; assim como o temperamento de ambos. Conclusão. Foram discutidos os resultados e suas implicações para estabelecer congruência entre o avanço conceitual e as evidencias empíricas sobre interações recíprocas

Análisis de la posible citotoxicidad asociada a las nanopartículas de selenio mediante herramientas de Proteómica cuantitativa

Comunicaciones a congreso

Long-Term Sustained Effect of Liver-Targeted Adeno-Associated Virus Gene Therapy for Mitochondrial Neurogastrointestinal Encephalomyopathy.

Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is caused by mutations in TYMP, the gene encoding the enzyme thymidine phosphorylase (TP). TP dysfunction results in systemic accumulation of the noxious TP substrates thymidine and deoxyuridine. Gene therapy using either a lentiviral vector or adeno-associated vector (AAV) has proven to be a feasible strategy, as both vectors restore biochemical homeostasis in a murine model of the disease. This study shows that the effect of an AAV containing the TYMP coding sequence transcriptionally targeted to the liver persists long term in mice. Although the vector copy number was diluted and AAV-mediated liver TP activity eventually reduced or lost after 21 months at the lowest vector doses, the effect was sustained (with a negligible decrease in TP activity) and fully effective on nucleoside homeostasis for at least 21 months at a dose of 2 × 1012 vg/kg. Macroscopic visual inspection of the animals' organs at completion of the study showed no adverse effects associated with the treatment. These results further support the feasibility of gene therapy for MNGIE

Efficacy of adeno-associated virus gene therapy in a MNGIE murine model enhanced by chronic exposure to nucleosides

Preclinical studies have shown that gene therapy is a feasible approach to treat mitochondrial neurogastrointestinal encephalomyopathy (MNGIE). However, the genetic murine model of the disease (Tymp/Upp1 double knockout, dKO) has a limited functional phenotype beyond the metabolic imbalances, and so the studies showing efficacy of gene therapy have relied almost exclusively on demonstrating correction of the biochemical phenotype. Chronic oral administration of thymidine (dThd) and deoxyuridine (dUrd) to dKO mice deteriorates the phenotype of the animals, providing a better model to test therapy approaches. dKO mice were treated with both dThd and dUrd in drinking water from weaning until the end of the study. At 8 - 11 weeks of age, mice were treated with several doses of adeno-associated virus (AAV) serotype 8 vector carrying the human TYMP coding sequence under the control of different liver-specific promoters (TBG, AAT, or HLP). The biochemical profile and functional phenotype were studied over the life of the animals. Nucleoside exposure resulted in 30-fold higher plasma nucleoside levels in dKO mice compared with non-exposed wild type mice. AAV-treatment provided elevated TP activity in liver and lowered systemic nucleoside levels in exposed dKO mice. Exposed dKO mice had enlarged brain ventricles (assessed by magnetic resonance imaging) and motor impairment (rotarod test); both were prevented by AAV treatment. Among all promoters tested, AAT showed the best efficacy. Our results show that AAV-mediated gene therapy restores the biochemical homeostasis in the murine model of MNGIE and, for the first time, demonstrate that this treatment improves the functional phenotype. This work was funded in part by the Spanish Instituto de Salud Carlos III, and the Generalitat de Catalunya. The disclosed funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Most mitochondrial dGTP is tightly bound to respiratory complex I through the NDUFA10 subunit

Biochemistry; Molecular medicineBioquímica; Medicina molecularBioquímica; Medicina molecularImbalanced mitochondrial dNTP pools are known players in the pathogenesis of multiple human diseases. Here we show that, even under physiological conditions, dGTP is largely overrepresented among other dNTPs in mitochondria of mouse tissues and human cultured cells. In addition, a vast majority of mitochondrial dGTP is tightly bound to NDUFA10, an accessory subunit of complex I of the mitochondrial respiratory chain. NDUFA10 shares a deoxyribonucleoside kinase (dNK) domain with deoxyribonucleoside kinases in the nucleotide salvage pathway, though no specific function beyond stabilizing the complex I holoenzyme has been described for this subunit. We mutated the dNK domain of NDUFA10 in human HEK-293T cells while preserving complex I assembly and activity. The NDUFA10E160A/R161A shows reduced dGTP binding capacity in vitro and leads to a 50% reduction in mitochondrial dGTP content, proving that most dGTP is directly bound to the dNK domain of NDUFA10. This interaction may represent a hitherto unknown mechanism regulating mitochondrial dNTP availability and linking oxidative metabolism to DNA maintenance.We thank Dr, Luke Formosa (Department of Biochemistry and Molecular Biology, Monash Biomedicine Discovery Institute, Monash University, Melbourne, Australia) for his valuable advice and assistance on NDUFA10 molecular studies and Dr. Francesc Canals and his team (Proteomics Laboratory, Vall d’Hebron Institute of Oncology [VHIO], Universitat Autònoma de Barcelona, Barcelona, Spain) for their assistance with LC-MS/MS analyses. This work was supported by the Spanish Ministry of Industry, Economy and Competitiveness [grants BFU2014-52618-R, SAF2017-87506, and PID2020-112929RB-I00 to Y.C.], by the Spanish Instituto de Salud Carlos III [grants PI21/00554 and PMP15/00025 to R.M.], co-financed by the European Regional Development Fund (ERDF), and by an NHMRC Project grant to M.R. (GNT1164459)

Identificación de los mecanismos moleculares asociados con la toxicidad inducida por MeHg mediante el análisis de la expresión proteica diferencial

Comunicaciones a congreso

Age-related metabolic changes limit efficacy of deoxynucleoside-based therapy in thymidine kinase 2-deficient mice

Thymidine kinase 2 (TK2) catalyses the phosphorylation of deoxythymidine (dThd) and deoxycytidine (dCtd) within mitochondria. TK2 deficiency leads to mtDNA depletion or accumulation of multiple deletions. In patients, TK2 mutations typically manifest as a rapidly progressive myopathy with infantile onset, leading to respiratory insufficiency and encephalopathy in the most severe clinical presentations. TK2-deficient mice develop the most severe form of the disease and die at average postnatal day 16. dThd+dCtd administration delayed disease progression and expanded lifespan of a knockin murine model of the disease. We daily administered TK2 knockout mice (Tk2 KO) from postnatal day 4 with equimolar doses of dThd+dCtd, dTMP+dCMP, dThd alone or dCtd alone. We monitored body weight and survival and studied different variables at 12 or 29 days of age. We determined metabolite levels in plasma and target tissues, mtDNA copy number in tissues, and the expression and activities of enzymes with a relevant role in mitochondrial dNTP anabolism or catabolism. dThd+dCtd treatment extended average lifespan of Tk2 KO mice from 16 to 34 days, attenuated growth retardation, and rescued mtDNA depletion in skeletal muscle and other target tissues of 12-day-old mice, except in brain. However, the treatment was ineffective in 29-day-old mice that still died prematurely. Bioavailability of dThd and dCtd markedly decreased during mouse development. Activity of enzymes catabolizing dThd and dCtd increased with age in small intestine. Conversely, the activity of the anabolic enzymes decreased in target tissues during mouse development. We also found that administration of dThd alone had the same impact on survival to that of dThd+dCtd, whereas dCtd alone had no influence on lifespan. dThd+dCtd treatment recruits alternative cytosolic salvage pathways for dNTP synthesis, suggesting that this therapy would be of benefit for any Tk2 mutation. dThd accounts for the therapeutic effect of the combined treatment in mice. During the first weeks after birth, mice experience marked tissue-specific metabolic regulations and ontogenetic changes in dNTP metabolism-related enzymes that limit therapeutic efficacy to early developmental stages. This study was funded by grants from the Spanish Ministry of Industry, Economy and Competitiveness, the Spanish Instituto de Salud Carlos III, the Fundación Inocente, Inocente, AFM Téléthon and the Generalitat de Catalunya. The disclosed funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript