Desarrollo de técnicas inmunoenzimáticas con anticuerpos recombinantes y de técnicas de PCR en tiempo real para la detección de almendra y nuez de Brasil de alimentos

La detección de frutos secos alergénicos en distintos tipos de alimentos es primordial para garantizar la salud de los individuos sensibles, ya que la ingesta inadvertida de pequeñas cantidades de estos alérgenos podría provocarles reacciones adversas e incluso la muerte. Las técnicas analíticas para su detección se basan en el análisis de proteínas o del ADN, pero no se disponía de métodos adecuados para la detección de todos los frutos secos alergénicos de declaración obligatoria en la Unión Europea. El objetivo global de esta tesis doctoral ha consistido en el desarrollo de novedosas técnicas de ELISA con anticuerpos recombinantes y de PCR en tiempo real que permitieran la detección sensible y específica de nuez de Brasil y almendra en alimentos. Este trabajo ha sido pionero en la obtención de nuevas sondas de afinidad para la detección de frutos secos alergénicos. Utilizando la tecnología de phage display y las genotecas Tomlinson I y J se han obtenido fago-anticuerpos de tipo scFv específicos frente a la nuez de Brasil y la almendra, con los que se han desarrollado técnicas de ELISA indirecto para la detección de estos alérgenos en alimentos, con límites de detección de 5,0 y 0,1 mg por gramo, respectivamente. Utilizando una aproximación proteómica se han identificado las globulinas 11S de la nuez de Brasil y de la almendra como las proteínas diana reconocidas por los anticuerpos recombinantes obtenidos frente a estas especies. Además, el análisis de numerosos productos comerciales tales como productos de panadería y bollería, galletas, chocolates, barritas energéticas, helados, cremas para untar y bebidas, mediante las técnicas de ELISA desarrolladas ha permitido descubrir errores o fraudes de etiquetado y detectar almendra y nuez de Brasil en productos en los que no se advierte de su presencia. La obtención de anticuerpos recombinantes frente a varios frutos secos de declaración obligatoria supone un avance importante desde el punto de vista metodológico, puesto que estas nuevas sondas de afinidad no requieren la utilización de animales de experimentación y constituyen una fuente ilimitada de anticuerpos específicos. Finalmente, una vez obtenidos los anticuerpos recombinantes se consiguió modificar mediante ingeniería genética los fragmentos scFv específicos frente a almendra y expresarlos en un sistema eucariota con el fin de producir scFv biotinilados in vivo. La estrategia desarrollada para la biotinilación in vivo de proteínas recombinantes empleando la levadura Pichia pastoris es una de las aportaciones más novedosas de esta tesis. Tras su purificación y multimerización con un núcleo de avidina-peroxidasa, los scFv biotinilados y utilizados en una técnica de ELISA conservaron la capacidad de detectar proteína de almendra hasta 470 mg por kg. En cuanto al desarrollo de técnicas genéticas, en esta tesis doctoral se han desarrollado técnicas de PCR en tiempo real para la detección de nuez de Brasil y almendra, que se encuentran entre las más sensibles publicadas en la bibliografía, con niveles de detección en alimentos de 2,5 y 0,1 mg por kg respectivamente, por lo que tienen una gran utilidad práctica para el control de alérgenos y la verificación del etiquetado, tanto para las autoridades sanitarias como para las industrias alimentarias. Los cebadores y sondas desarrollados han permitido analizar un gran número de alimentos comerciales y poner de manifiesto la presencia de frutos secos alergénicos no declarados en un elevado porcentaje de muestras, que oscila entre un 10 por ciento en el caso de la nuez de Brasil y un 40 por ciento en el caso de la almendra. Puede concluirse que las técnicas desarrolladas en este trabajo proporcionan herramientas específicas, rápidas y sensibles para detectar la presencia de nuez de Brasil y almendra, contribuyendo al cumplimiento de la normativa europea en materia de etiquetado de los alimentos

Preliminary Investigation on the Physicochemical and Functional Properties of Commercial Salmorejo Found in Spanish Supermarkets

Salmorejo is a traditional Spanish food made of raw tomatoes, bread, garlic, and virgin olive oil. The food industry aims to satisfy consumer demand for ready-to-eat salmorejo while trying to maintain characteristics of the homemade product. In this work, we have assessed physical-chemical and color parameters, radical scavenging activity, and total polyphenol, lycopene, and β-carotene contents of raw and pasteurized commercial salmorejo, as well as homemade (raw) salmorejo samples. Our results showed that heat treatment had a significant influence on color parameters of salmorejo, with pasteurized samples being less red and exhibiting an increase in the degree of browning. Pasteurized samples also showed the highest radical scavenging activity when expressing the results per dry weight. However, when water content was considered, radical scavenging activity was superior in homemade samples when analyzing non-polar molecules extracted with acetone. Results were similar for polyphenol content. It was also observed that heat treatment affected lycopene but not β-carotene content. When acquiring commercial salmorejo at the supermarket, consumers have the option to choose between already prepared pasteurized or raw salmorejo. According to the results obtained in this work, physicochemical and functional properties of commercial raw salmorejo were comparable to a larger extent than pasteurized salmorejo compared to those exhibited by homemade salmorejo samples

Coenzyme Q10 and Cardiovascular Diseases

Coenzyme Q10 (CoQ10), which plays a key role in the electron transport chain by providing an adequate, efficient supply of energy, has another relevant function as an antioxidant, acting in mitochondria, other cell compartments, and plasma lipoproteins. CoQ10 deficiency is present in chronic and age-related diseases. In particular, in cardiovascular diseases (CVDs), there is a reduced bioavailability of CoQ10 since statins, one of the most common lipid-lowering drugs, inhibit the common pathway shared by CoQ10 endogenous biosynthesis and cholesterol biosynthesis. Different clinical trials have analyzed the effect of CoQ10 supplementation as a treatment to ameliorate these deficiencies in the context of CVDs. In this review, we focus on recent advances in CoQ10 supplementation and the clinical implications in the reduction of cardiovascular risk factors (such as lipid and lipoprotein levels, blood pressure, or endothelial function) as well as in a therapeutic approach for the reduction of the clinical complications of CVD

Twitter as an educational tool for autonomous and collaborative learning in the Veterinary Degree

Desde el punto de vista educativo, las redes sociales facilitan el intercambio de una gran cantidad de información especializada en tiempo real. Twitter permite el desarrollo de habilidades comunicativas y la capacidad de síntesis por su sencilla y directa forma de comunicación. Nuestro objetivo ha sido determinar el valor de Twitter como instrumento educativo de aprendizaje autónomo y colaborativo. Para ello, se creó una cuenta para la asignatura optativa “el Animal de Laboratorio”, perteneciente al Grado de Veterinaria. Se llevaron a cabo dos actividades; i) la gestión temporal de la cuenta por parte de distintos grupos de trabajo y, ii) el seguimiento e interacción con la cuenta desde los perfiles individuales de cada alumno. Se evaluó mediante cuestionarios tanto la situación inicial del alumnado, así como los logros finales conseguidos. Los resultados obtenidos destacan su funcionalidad como instrumento de aprendizaje autónomo. Además, nos muestran que es una metodología que fomenta el aprendizaje colaborativo, permitiendo interactuar, compartir conclusiones y establecer un reparto de tareas. Igualmente, su uso aumenta el interés y la motivación, al igual que promueve la participación del alumnado. En definitiva, se pone de manifiesto la idoneidad de Twitter como herramienta educativa en el proceso de enseñanza-aprendizaje competencial actual.From an educational standpoint, the use of social networks facilitates the exchange of information, allowing access to a large amount of specialized information in real time. The social network Twitter is characterized by its user-friendly and straightforward way of social interaction. The main objective of this project has been to determine the value of Twitter as an educational tool for autonomous and collaborative learning. To that end, a Twitter account was made for the elective subject "Laboratory Animal", belonging to the Veterinary Degree. Two activities were carried out: i) temporary management of the account profile by different working groups, and ii) interaction with the account profile using students private accounts. The starting point of students, as well as the final achievements regarding the stated objective, were evaluated using questionnaires. It stands out the functionality of the tool as an autonomous learning instrument, as it allows students to be conscious of their learning process. It also came across as a methodology that encourages collaborative learning, as it promotes interaction, sharing conclusions and teamwork building. Finally, it increases interest and motivation, as well as participation. In conclusion, Twitter migh be a reliable tool to facilitate competency-based learning in superior education

Endothelial Dysfunction and Advanced Glycation End Products in Patients with Newly Diagnosed Versus Established Diabetes: From the CORDIOPREV Study

Endothelial dysfunction and intima-media thickness of common carotid arteries (IMT-CC) are considered subclinical markers of atherosclerotic cardiovascular disease (ASCVD). Advanced glycation end products (AGEs) are increased in type 2 diabetes mellitus (T2DM) patients, compared with non-diabetics, being implicated in micro- and macrovascular complications. Our aim was to compare serum AGEs levels and subclinical atherosclerotic markers between patients with established and newly diagnosed T2DM. Among 540 patients with T2DM and coronary heart disease from the CORDIOPREV study, 350 patients had established T2DM and 190 patients had newly diagnosed T2DM. Serum levels of AGEs (methylglyoxal (MG) and N-carboxymethyl lysine (CML)) and subclinical atherosclerotic markers (brachial flow-mediated vasodilation (FMD) and IMT-CC) were measured. AGEs levels (all p < 0.001) and IMT-CC (p = 0.025) were higher in patients with established vs. newly diagnosed T2DM, whereas FMD did not differ between the two groups. Patients with established T2DM and severe endothelial dysfunction (i.e., FMD < 2%) had higher serum MG levels, IMT-CC, HOMA-IR and fasting insulin levels than those with newly diagnosed T2DM and non-severe endothelial dysfunction (i.e., FMD ≥ 2%) (all p < 0.05). Serum CML levels were greater in patients with established vs. newly diagnosed T2DM, regardless of endothelial dysfunction severity. Serum AGEs levels and IMT-CC were significantly higher in patients with established vs. newly diagnosed T2DM, highlighting the progressively increased risk of ASCVD in the course of T2DM. Establishing therapeutic strategies to reduce AGEs production and delay the onset of cardiovascular complications in newly diagnosed T2DM patients or minimize ASCVD risk in established T2DM patients is needed

Quality and Quantity of Protein Intake Influence Incidence of Type 2 Diabetes Mellitus in Coronary Heart Disease Patients: From the CORDIOPREV Study

Evidence suggests that enriching a diet with plant-based proteins could reduce the risk of developing type 2 diabetes mellitus. In the present work, we evaluated the association between the change in plant protein intake (adjusted by energy) and incidence of type 2 diabetes mellitus in patients with coronary heart disease from the CORDIOPREV (coronary diet intervention with olive oil and cardiovascular prevention) study. At baseline and during the follow-up, patients underwent medical examination and blood and oral glucose tolerance tests. Information on patient’s dietary intake was gathered by registered dietitians using a validated food frequency questionnaire. A total of 106 out of 436 nondiabetic patients at baseline developed type 2 diabetes mellitus after a median follow-up of 60 months. Cox regression analyses showed that patients who belonged to the group that increased plant protein intake exhibited a lower risk of developing the disease (HR = 0.64, (0.43–0.96)). Changes in plant protein intake were positively correlated with changes in carbohydrates, fibre, and legumes intake and negatively correlated with changes in saturated fatty acids intake. Results of the present study support the need of improving diet with plant-based proteins to prevent the onset of type 2 diabetes mellitus

Beta cell functionality and hepatic insulin resistance are major contributors to type 2 diabetes remission and starting pharmacological therapy: from CORDIOPREV randomized controlled trial

In order to assess whether previous hepatic IR (Hepatic-IR fasting) and beta-cell functionality could modulate type 2 diabetes remission and the need for starting glucose- lowering treatment, newly-diagnosed type 2 diabetes participants who had never received glucose-lowering treatment (190 out of 1002) from the CORonary Diet Intervention with Olive oil and cardiovascular PREVention study (a prospective, randomized and controlled clinical trial), were randomized to consume a Mediterranean or a low-fat diet. Type 2 diabetes remission was defined according to the American Diabetes Association recommendation for levels of HbA1c, fasting plasma glucose and 2h plasma glucose after oral glucose tolerance test, and having maintained them for at least 2 consecutive years. Patients were classified according to the median of Hepatic-IR fasting and beta-cell functionality, measured as the disposition index (DI) at baseline. Cox proportional hazards regression determined the potential for Hepatic-IR fasting and DI indexes as predictors of diabetes remission and the probability of starting pharmacological treatment after a 5-year follow-up. Low-Hepatic-IR fasting or high-DI patients had a higher probability of diabetes remission than high-Hepatic-IR fasting or low-DI subjects (HR:1.79; 95% CI 1.06_3.05; and HR:2.66; 95% CI 1.60_4.43, respectively) after a dietary intervention with no pharmacological treatment and no weight loss. The combination of low- Hepatic-IR fasting and high-DI presented the highest probability of remission (HR:4.63; 95% CI 2.00_10.70). Among patients maintaining diabetes, those with high- Hepatic-IR fasting and low-DI showed the highest risk of starting glucose-lowerin

Recent advances in food allergy

Food allergy is a public health issue that has significantly increased worldwide in the past decade, affecting consumers’ quality of life and making increasing demands on health service resources. Despite recent advances in many areas of diagnosis and treatment, our general knowledge of the basic mechanisms of the disease remain limited i.e., not at pace with the exponential number of new cases and the explosion of new technologies. Many important key questions remain: What defines a major allergen? Why do some individuals develop food allergies and others do not? Which are the environmental factors? Could the environmental factors be monitored through epigenetics or modified by changes in the microbiome? Can tolerance to food be induced? Why are some foods more likely to trigger allergies than others? Does the route and timing of exposure have any role on sensitization? These and many other related questions remain unanswered. In this short review some of these topics are addressed in the light of recent advances in the area

Recent advances in food allergy

Summary Food allergy is a public health issue that has significantly increased worldwide in the past decade, affecting consumers’ quality of life and making increasing demands on health service resources. Despite recent advances in many areas of diagnosis and treatment, our general knowledge of the basic mechanisms of the disease remain limited i.e., not at pace with the exponential number of new cases and the explosion of new technologies. Many important key questions remain: What defines a major allergen? Why do some individuals develop food allergies and others do not? Which are the environmental factors? Could the environmental factors be monitored through epigenetics or modified by changes in the microbiome? Can tolerance to food be induced? Why are some foods more likely to trigger allergies than others? Does the route and timing of exposure have any role on sensitization? These and many other related questions remain unanswered. In this short review some of these topics are addressed in the light of recent advances in the area

Uso de Twitter como herramienta educativa de aprendizaje autónomo y colaborativo en el Grado de Veterinaria

Desde el punto de vista educativo, el uso de redes sociales facilita el intercambio de información, haciéndolo más ágil y accesible y permitiendo el acceso a gran cantidad de información especializada en tiempo real. Twitter se caracteriza por su sencilla y directa forma de comunicación, permitiendo el desarrollo de habilidades comunicativas y la capacidad de síntesis. El objetivo principal de este proyecto ha sido determinar el valor de la red social Twitter como instrumento educativo de aprendizaje autónomo y colaborativo. Se creó una cuenta de Twitter para la asignatura optativa de “el Animal de Laboratorio”, perteneciente al Grado de Veterinaria (@cuartaErre). Se llevaron a cabo dos actividades: i) la gestión temporal del propio perfil por parte de distintos grupos de trabajo, y ii) el seguimiento e interacción con el perfil @cuartaErre desde cuentas individuales de cada alumno. Se evaluó mediante cuestionarios tanto la situación inicial del alumnado, como los logros finales respecto al objetivo planteado, siguiendo una metodología descriptiva. Destaca su funcionalidad como instrumento de aprendizaje autónomo, permitiendo al estudiantado reflexionar acerca de su propio proceso de aprendizaje. Se trata, además, de una metodología que fomenta el aprendizaje colaborativo, permitiendo interactuar, compartir conclusiones y establecer un reparto de tareas. Igualmente, aumenta el interés y la motivación, al igual que promueve la participación. En definitiva, se pone de manifiesto la idoneidad del uso de Twitter como herramienta educativa en un proceso de enseñanza-aprendizaje competencial como el actual