Evaluating the pharmacological response in fluorescence microscopy images: The Δm algorithm

Current drug discovery procedures require fast and effective quantification of the pharmacological response evoked in living cells by agonist compounds. In the case of G-protein coupled receptors (GPCRs), the efficacy of a particular drug to initiate the endocytosis process is related to the formation of endocytic vesicles or endosomes and their subsequent internalisation within intracellular compartments that can be observed with high spatial and temporal resolution by fluorescence microscopy techniques. Recently, an algorithm has been proposed to evaluate the pharmacological response by estimating the number of endosomes per cell on time series of images. However, the algorithm was limited by the dependence on some manually set parameters and in some cases the quality of the image does not allow a reliable detection of the endosomes. Here we propose a simple, fast and automated image analysis method?the ?m algorithm- to quantify a pharmacological response with data obtained from fluorescence microscopy experiments. This algorithm does not require individual object detection and computes the relative increment of the third order moment in fluorescence microscopy images after filtering with the Laplacian of Gaussian function. It was tested on simulations demonstrating its ability to discriminate different experimental situations according to the number and the fluorescence signal intensity of the simulated endosomes. Finally and in order to validate this methodology with real data, the algorithm was applied to several time-course experiments based on the endocytosis of the mu opioid receptor (MOP) initiated by different agonist compounds. Each drug displayed a different ?m sigmoid time-response curve and statistically significant differences were observed among drugs in terms of efficacy and kinetic parameters

Detection, size characterization and quantification of silver nanoparticles in consumer products by particle collision coulometry

Silver nanoparticles (AgNPs) are widely used in industrial and consumer products owing to its antimicrobial nature and multiple applications. Consequently, their release into the environment is becoming a big concern because of their negative impacts on living organisms. In this work, AgNPs were detected at a potential of + 0.70 V vs. Ag/AgCl reference electrode, characterized, and quantified in consumer products by particle collision coulometry (PCC). The electrochemical results were compared with those measured with electron microscopy and single-particle inductively coupled plasma mass spectrometry. The theoretical and practical peculiarities of the application of PCC technique in the characterization of AgNPs were studied. Reproducible size distributions of the AgNPs were measured in a range 10–100 nm diameters. A power allometric function model was found between the frequency of the AgNPs collisions onto the electrode surface and the number concentration of nanoparticles up to a silver concentration of 1010 L-1 (ca. 25 ng L-1 for 10 nm AgNPs). A linear relationship between the number of collisions and the number concentration of silver nanoparticles was observed up to 5 × 107 L-1. The PCC method was applied to the quantification and size determination of the AgNPs in three-silver containing consumer products (a natural antibiotic and two food supplements). The mean of the size distributions (of the order 10–20 nm diameters) agrees with those measured by electron microscopy. Graphical abstract: [Figure not available: see fulltext.]

Cuantificación de ácidos grasos a partir de biomasa microalgal

Long chain polyunsaturated fatty acids are focusing the interest of researchers as long as analytical techniques are being improved. The Lepage and Roy direct transesterification method is being widely used to measure fatty acid content of microalgae which are proposed as potential sources of these products. In this paper, the physical state of the biomass to be analyzed is studied. Comparison has been stated between lyophilized biomass and wet biomass just harvested. Furthermore, the amount of sample and the number of methyl ester extractions have also been studied. From the statistical analysis carried out, wet biomass to fatty acid determination and the adequate range of sample are proposed. Three methyl ester extractions improve the resolution of GC analysis.El interés creciente por los ácidos grasos poliinsaturados de cadena larga viene favorecido por la mejora de las técnicas de análisis. La metodología de transesterificación directa propuesta por Lepage y Roy está siendo ampliamente utilizada para la determinación del contenido en ácidos grasos de microalgas, consideradas como potenciales fuentes de estos productos. En el presente trabajo, se analiza la influencia del estado físico de la biomasa analizada, seca, normalmente por liofilización, o húmeda recién cosechada del cultivo. Asimismo, se analiza el efecto de la cantidad de biomasa empleada y del número de extracciones de los esteres metílicos obtenidos. Del estudio estadístico realizado se pone de manifiesto la importancia del estado físico de la muestra, aconsejándose el uso de biomasa directamente obtenida del cultivo. Se proponen los intervalos de cantidad de biomasa para metilación, así como la conveniencia de realizar tres extracciones previas a la inyección en el cromatógrafo, que favorece la resolución del análisis

Efectos de una unidad didáctica de mimo basada en el modelo de Educación Deportiva sobre la interculturalidad

The aim of this study was to assess the impact of a Sport Education unit on elementary school students’ intercultural competence. The program involved the participation of 28 fifth-grade students (M = 10.75; DT = .75) from a state school in Region of Murcia, Spain.A quasi-experimental design, pretest and posttest (of quantitative and qualitative nature) measures, was used. The data included students’ questionnaires and interviews, andteachers’ interviews and diary notes. Results indicated that students improved on positive emotional responses, as well as providing help and having better relations with their peers from different cultures/countries. The incidence of methodological elements, communication and the social climate of the classroom on the development of intercultural competence is also discussed. Lines of future research are suggested in order to improve the model for the development of intercultural competence.El propósito de este estudio fue conocer el impacto generado por una unidad didáctica de mimo basada en el Modelo de Educación Deportiva (MED) sobre la competencia intercultural del alumnado de primaria. Participaron 28 estudiantes (M = 10.75; DT = .75) de quinto curso de educación primaria pertenecientes a un colegio ubicado en la Región de Murcia (España). Se desarrolló un diseño cuasiexperimental con medidas pretest y postest, de naturaleza cuantitativa y cualitativa. La información fue recabada mediante cuestionarios y entrevistas al alumnado, así como diarios y entrevistas a los docentes. Los resultados mostraron mejoras en la respuesta emocional positiva, así como en la ayuda, y las relaciones entre iguales de distintas culturas y países. Se discute también la incidencia de los elementos metodológicos, la comunicación y el clima social del aula sobre el desarrollo de la competencia intercultural. Se sugieren líneas de investigación futura al objeto de perfeccionar el modelo para el desarrollo de la competencia intercultural.

Functionalization using biocompatible carboxylated cyclodextrins of iron-based nanoMIL-100

Here we report the first example of nanoMIL-100 particles modified with monomeric cyclodextrin derivatives of different length by exploiting strong interactions between non-saturated iron trimers at the external surface and carboxylate functionalities located at the end of biocompatible and flexible linkers of cyclodextrins. The main results revealed that, after the functionalization, the cyclodextrins are selectively located at the external surfaces covering the nanoparticles. Z potential measurements show that this functionalization induced changes respect to the bare nanoMIL-100 particles, however, the presence of the cyclodextrins does not modify the size neither porosity of the nanoparticles. The amount of cyclodextrins attached, investigated by thermogravimetry, increases with the length of the linker between CD cavity and nanoparticle surface, reaching up a 9 % wt. Auger spectroscopy suggested a clear predominant sp3 character after the functionalizations (vs. sp2 predominance in the unmodified nanoMIL-100). This study supposes the creation of an alternative family of hybrids based on carboxylated monomeric cyclodextrins

El modelo de Educación Deportiva y la orientación. Efectos en la satisfacción con la vida, las inteligencias múltiples, las necesidades psicológicas básicas y las percepciones sobre el modelo de los adolescentes

The aim of this study was to assess the effects of a Sport Education (SE) model-based teaching unit through orienteering on grade 9 (14-17 years) students’ life satisfaction, multiple intelligences, basic psychological needs and curricular model perceptions.81 students (Mage = 14.62; SD = .80) allocated in three intact groups from a high-school in Region of Murcia (Spain), took part in this unit. Based on a quasi-experimental design (pretest-posttest measures), data were collected from interviews (teacher perspective) and both quantitative and qualitative questionnaires (student perspective). Significant improvements were obtained on the development of the three main SE goals (competency, literacy and enthusiasm), whereas no significant changes were found on life satisfaction and multiple intelligences. Findings confirmed orienteering as a suitable content to be taught according through the pedagogical principles of SE model.Likewise, results showed its benefits on the development of the objectives this pursues.Se examinaron los efectos de una unidad didáctica de orientación deportiva (OD) basada en el modelo de Educación Deportiva (ED) sobre la satisfacción con la vida, las inteligencias múltiples, las necesidades psicológicas básicas y las percepciones del modelo en estudiantesde tercer curso de educación secundaria. La muestra estuvo comprendida por 81 estudiantes (M de edad = 14.62; DT = .80) distribuidos en tres grupos naturales, pertenecientes a un instituto ubicado en la Región de Murcia (España). A partir de un diseño cuasi-experimental (medidas pretest-postest), se recabaron datos mediante entrevistas (percepción docente), cuestionarios cuantitativos y cualitativos (percepción discente). Se obtuvieronmejoras significativas en el desarrollo de los tres objetivos que persigue el modelo (competencia, cultura deportiva y entusiasmo), al tiempo queno se revelaron cambios significativos en la satisfacción con la vida y en las inteligencias múltiples. Los resultados constatan a la OD como contenido apto para ser abordado mediante los principios del modelo de ED y respaldansu eficaciarespecto a los objetivos que este últimopersigue

Mortality and cardiovascular disease burden of uncontrolled diabetes in a registry-based cohort: the ESCARVAL-risk study

BACKGROUND: Despite the epidemiological evidence about the relationship between diabetes, mortality and cardiovascular disease, information about the population impact of uncontrolled diabetes is scarce. We aimed to estimate the attributable risk associated with HbA1c levels for all-cause mortality and cardiovascular hospitalization. METHODS: Prospective study of subjects with diabetes mellitus using electronic health records from the universal public health system in the Valencian Community, Spain 2008-2012. We included 19,140 men and women aged 30 years or older with diabetes who underwent routine health examinations in primary care. RESULTS: A total of 11,003 (57%) patients had uncontrolled diabetes defined as HbA1c ≥6.5%, and, among those, 5325 participants had HbA1c ≥7.5%. During an average follow-up time of 3.3 years, 499 deaths, 912 hospitalizations for coronary heart disease (CHD) and 786 hospitalizations for stroke were recorded. We observed a linear and increasingly positive dose-response of HbA1c levels and CHD hospitalization. The relative risk for all-cause mortality and CHD and stroke hospitalization comparing patients with and without uncontrolled diabetes was 1.29 (95 CI 1.08,1.55), 1.38 (95 CI 1.20,1.59) and 1.05 (95 CI 0.91, 1.21), respectively. The population attributable risk (PAR) associated with uncontrolled diabetes was 13.6% (95% CI; 4.0-23.9) for all-cause mortality, 17.9% (95% CI; 10.5-25.2) for CHD and 2.7% (95% CI; - 5.5-10.8) for stroke hospitalization. CONCLUSIONS: In a large general-practice cohort of patients with diabetes, uncontrolled glucose levels were associated with a substantial mortality and cardiovascular disease burden

Dry selection and wet evaluation for the rational discovery of new anthelmintics

Helminths infections remain a major problem in medical and public health. In this report, atom-based 2D bilinear indices, a TOMOCOMD-CARDD (QuBiLs-MAS module) molecular descriptor family and linear discriminant analysis (LDA) were used to find models that differentiate among anthelmintic and non-anthelmintic compounds. Two classification models obtained by using non-stochastic and stochastic 2D bilinear indices, classified correctly 86.64% and 84.66%, respectively, in the training set. Equation 1(2) correctly classified 141(135) out of 165 [85.45%(81.82%)] compounds in external validation set. Another LDA models were performed in order to get the most likely mechanism of action of anthelmintics. The model shows an accuracy of 86.84% in the training set and 94.44% in the external prediction set. Finally, we carry out an experiment to predict the biological profile of our ‘in-house’ collections of indole, indazole, quinoxaline and cinnoline derivatives (∼200 compounds). Subsequently, we selected a group of nine of the theoretically most active structures. Then, these chemicals were tested in an invitro assay and one good candidate (VA5-5c) as fasciolicide compound (100% of reduction at concentrations of 50 and 10 mg/L) was discovered.Yovani Marrero-Ponce acknowledges the support from USFQ with partial finance of project ID5455Peer Reviewe