Combining data acquisition modes in liquid-chromatography–tandem mass spectrometry for comprehensive determination of acylcarnitines in human serum

Acylcarnitines (ACs) are metabolites involved in fatty acid β-oxidation and organic acid metabolism. Metabolic disorders associated to these two processes can be evaluated by determining the complete profile of ACs. In this research, we present an overall strategy for identification, confirmation, and quantitative determination of acylcarnitines in human serum. By this strategy we identified the presence of 47 ACs from C2 to C24 with detection of the unsaturation degree by application of a data-independent acquisition (DIA) liquid chromatography–tandem mass spectrometry (LC–MS/MS) method. Complementary, quantitative determination of ACs is based on a high-throughput and fully automated method consisting of solid-phase extraction on-line coupled to LC–MS/MS in data-dependent acquisition (DDA) to improve analytical features avoiding the errors associated to sample processing. Quantitation limits were at pg mL–1 level, the intra-day and between-day variability were below 15–20%, respectively; and the accuracy, expressed as bias, was always within ± 25%. The proposed method was tested with 40 human volunteers to determine the relative concentration of ACs in serum and identify predominant forms. Significant differences were detected by comparing the ACs profile of obese versus non-obese individuals

Strategies for proteomic analysis of blood glycated proteins

Comunicaciones a congreso

Quantitative analysis of protein glycation in clinical samples

Comunicaciones a congreso

Comparison of drying techniques for extraction of bioactive compounds from olive-tree materials

Olive tree vegetal materials are considered a powerful source for the isolation of bioactive compounds—mainly phenols and triterpenic acids. However, the high humidity content of them reduces their preservation and extractability to a liquid solvent. Accordingly, a drying step is crucial to homogenize the material and to obtain an efficient extraction. We studied the influence of the drying process on the extraction efficiency of bioactive compounds from olive vegetal material. For this purpose, we evaluated the effects of four drying processes on the solid–liquid extraction of bioactive compounds from two by-products, olive leaves and pomace, and olive fruits harvested from two cultivars, Alfafara and Koroneiki. Infrared-assisted drying (IAD) was the most suited approach to obtain extracts enriched in oleuropein from leaves (28.5 and 22.2% dry weight in Alfafara and Koroneiki, respectively). In the case of pomace, lyophilization and microwave-assisted drying led to extracts concentrated in oleacein and oleuropein aglycone, whereas IAD and oven-drying led to extracts with enhanced contents of hydroxytyrosol glucoside and hydroxytyrosol, respectively. The drying process considerably affected the chemical composition of extracts obtained from fruits. Changes in the composition of the extracts were explained essentially by the drying process conditions using auxiliary energies, temperature, and time, which promoted chemical alterations and increased the extractability of the compounds. Therefore, the drying protocol should be selected depending on the phenolic content and initial raw material

Glycated platelets proteome

Comunicaciones a congreso

Development of a method for the evaluation of comptences acquisition and its comparison with the tradicional evaluation system in subjects of Chemistry Degree

Un aspecto diferencial de la enseñanza de Grado en el Espacio Europeo de Enseñanza Superior (EEES), recogido en el Marco Español de Cualificaciones (MECES), es la definición de programas formativos que implican conocimientos y competencias. En este trabajo se han preparado rúbricas para evaluar las competencias a adquirir por el alumnado en dos asignaturas del Grado en Química. Las rúbricas diseñadas permiten dividir el conjunto de competencias en elementos diferenciados que se pueden evaluar diseñando y seleccionando actividades apropiadas. En esta herramienta se definen indicadores escalados para determinar el nivel alcanzado en cada competencia. Los resultados obtenidos con la aplicación de las rúbricas han sido correlacionados con los obtenidos en la evaluación habitual. Se propone así un método de trabajo flexible y dinámico para mejorar el proceso de evaluación.A differential aspect of the teaching of Degree in the European Higher Education Area (EHEA), included in the Spanish Qualifications Framework (MECES), is the definition of training programs that involve knowledge and skills. In this paper, rubrics have been prepared to evaluate the competences to be acquired by the students in two subjects of the Degree in Chemistry. The designed rubrics allow dividing the set of competences into differentiated elements that can be evaluated by designing and selecting appropriate activities. In this tool, scaled indicators are defined to determine the level reached in each competence. The results obtained with the application of the rubrics has been correlated with those obtained in the usual evaluation. A work method that is flexible and dynamic to improve the evaluation process is proposed

Promotion of self-employed work in the laboratory practices of students of the degree in chemistry: case study

En este estudio se ha perseguido incrementar la participación de los alumnos en las prácticas de laboratorio. El motivo principal ha sido evitar que el alumno memorice los conceptos teóricos sin ser capaz de aplicarlos a la resolución de problemas reales, y así adquiera compentencias útiles para cuando se introduzca en el mundo laboral. La nueva metodología se desarrolló en una asignatura optativa del Grado de Química. Para abordar la nueva metodología propuesta en este proyecto, los alumnos elaboraron los protocolos de tres prácticas, sobre tres temas propuestos por el profesorado. Una vez supervisados dichos protocolos por el profesorado, los alumnos las realizaron de forma autónoma en el laboratorio. La evaluación de los alumnos se realizó mediante: elaboración de los protocolos de prácticas, realización de las prácticas de laboratorio (mediante rúbrica) y cuestionario sobre conceptos teóricos (mediante la herramienta Kahoot). Finalmente, se evaluó el grado de aceptación de la nueva metodología docente por parte del alumnado, para detectar aspectos a mejorar para el próximo año académico. Esta metodología ha permitido una mayor implicación del alumnado en las sesiones de laboratorio, ya que pudieron aplicar los conceptos teóricos adquiridos sobre Química Analítica y adquirir competencias, tales como capacidad de aprendizaje autónomo.In this study we have sought to increase the participation of students in laboratory practices. The main reason has been to prevent the student from memorizing the theoretical concepts without being able to apply them to the resolution of real problems, and thus acquire useful competences for when entering into the working world. The new methodology was developed in an optional subject of the Degree in Chemistry. Therefore, it was proposed that they themselves elaborate the protocols of practices, on subjects proposed by the teaching staff, and that they carried them out independently in the laboratory. The evaluation of the students' work was carried out through three activities: preparation of the practical protocols, carrying out of the laboratory practices (using a rubric) and questionnaire on theoretical concepts (using the Kahoot tool). Finally, the degree of acceptance of the new teaching methodology by the students was evaluated. This allowed to detect the aspects to improve for the next academic year. This methodology has allowed a greater involvement of the students in the laboratory sessions, since they could apply the theoretical concepts acquired on Analytical Chemistry and acquire competences, such as autonomous learning capacity

Evaluation of Antioxidant and Wound-Healing Properties of EHO-85, a Novel Multifunctional Amorphous Hydrogel Containing Olea europaea Leaf Extract

The excess of free radicals in the wound environment contributes to its stagnation during the inflammatory phase, favoring hard-to-heal wounds. Oxidative stress negatively affects cells and the extracellular matrix, hindering the healing process. In this study, we evaluated the antioxidant and wound-healing properties of a novel multifunctional amorphous hydrogel-containing Olea europaea leaf extract (OELE). Five assessments were performed: (i) phenolic compounds characterization in OELE; (ii) absolute antioxidant activity determination in OELE and hydrogel (EHO-85); (iii) antioxidant activity measurement of OELE and (iv) its protective effect on cell viability on human dermal fibroblasts (HDFs) and keratinocytes (HaCaT); and (v) EHO-85 wound-healing-capacity analysis on diabetic mice (db/db; BKS.Cg-m+/+Leprdb). The antioxidant activity of OELE was prominent: 2220, 1558, and 1969 µmol TE/g by DPPH, ABTS, and FRAP assays, respectively. Oxidative stress induced with H2O2 in HDFs and HaCaT was normalized, and their viability increased with OELE co-treatment, thus evidencing a protective role. EHO-85 produced an early and sustained wound-healing stimulating effect superior to controls in diabetic mice. This novel amorphous hydrogel presents an important ROS scavenger capacity due to the high phenolic content of OELE, which protects skin cells from oxidative stress and contributes to the physiological process of wound healing.This research was funded by QUESPER R&D, and partially by INNCORPORA-TU-2011-1886 subprogramme (Ministry of Economy and Competitiveness, Spain) and the programme for the Reinforcement of Research Activity in the Clinical Management Units of the Andalusian Health Service (Department of Health. Regional Government of Andalusia, Spain)

Early Salmonella Typhimurium infection in pigs disrupts Microbiome composition and functionality principally at the ileum mucosa

peer-reviewedSalmonella is a major foodborne pathogen which successfully infects animal species for human consumption such as swine. The pathogen has a battery of virulence factors which it uses to colonise and persist within the host. The host microbiota may play a role in resistance to, and may also be indirectly responsible from some of the consequences of, Salmonella infection. To investigate this, we used 16S rRNA metagenomic sequencing to determine the changes in the gut microbiota of pigs in response to infection by Salmonella Typhimurium at three locations: ileum mucosa, ileum content and faeces. Early infection (2 days post-infection) impacted on the microbiome diversity at the mucosa, reflected in a decrease in representatives of the generally regarded as desirable genera (i.e., Bifidobacterium and Lactobacillus). Severe damage in the epithelium of the ileum mucosa correlated with an increase in synergistic (with respect to Salmonella infection; Akkermansia) or opportunistically pathogenic bacteria (Citrobacter) and a depletion in anaerobic bacteria (Clostridium spp., Ruminococcus, or Dialliser). Predictive functional analysis, together with metabolomic analysis revealed changes in glucose and lipid metabolism in infected pigs. The observed changes in commensal healthy microbiota, including the growth of synergistic or potentially pathogenic bacteria and depletion of beneficial or competing bacteria, could contribute to the pathogen’s ability to colonize the gut successfully. The findings from this study could be used to form the basis for further research aimed at creating intervention strategies to mitigate the effects of Salmonella infection