Semipermeable capsules wrapping a multifunctional and self-regulated co-culture microenvironment for osteogenic differentiation

A new concept of semipermeable reservoirs containing co-cultures of cells and supporting microparticles is presented, inspired by the multi-phenotypic cellular environment of bone. Based on the deconstruction of the â stem cell nicheâ , the developed capsules are designed to drive a self-regulated osteogenesis. PLLA microparticles functionalized with collagen I, and a co-culture of adipose stem (ASCs) and endothelial (ECs) cells are immobilized in spherical liquified capsules. The capsules are coated with multilayers of poly(L-lysine), alginate, and chitosan nano-assembled through layer-by-layer. Capsules encapsulating ASCs alone or in a co-culture with ECs are cultured in endothelial medium with or without osteogenic differentiation factors. Results show that osteogenesis is enhanced by the co-encapsulation, which occurs even in the absence of differentiation factors. These findings are supported by an increased ALP activity and matrix mineralization, osteopontin detection, and the up regulation of BMP-2, RUNX2 and BSP. The liquified co-capsules also act as a VEGF and BMP-2 cytokines release system. The proposed liquified capsules might be a valuable injectable self-regulated system for bone regeneration employing highly translational cell sources.

Hypertension and clustering of cardiovascular risk factors in a community in Southeast Brazil: the Bambuí Health and Ageing Study

OBJECTIVE - A population-based prospective study was analysed to: a) determine the prevalence of hypertension; b) investigate the clustering of other cardiovascular risk factors and c) verify whether older differed from younger adults in the pattern of clustering. METHODS - The data comprised a representative sample of the population of Bambuí, Brazil. Multiple logistic regression was used to investigate the independent association between hypertension and selected factors. RESULTS - A total of 820 younger adults (82.5%) and 1494 older adults (85.9%) participated in this study. The overall prevalence of hypertension was 24.8% (SE=1.4 %), being higher in women (26.9±1.5%) than in men (22.0± 1.7%) (p=0.033). Hypertension was positively and significantly associated with physical inactivity, overweight, hypercholesterolemia hyperglycemia and hypertriglyceridemia. The coexistence of hypertension with 4 or more of these risk factors occurred 6 times more than expected by chance, after adjusting for age and sex (OR=6.3; 95%CI: 3.4-11.9). The pattern of risk factor clustering in hypertensive individuals differed with age. CONCLUSION - Our results reinforce the need to increase detection and treatment of hypertension and to approach patients' global risk profiles.

Electronic tongue: a versatile tool for mineral and fruit-flavored waters recognition

Natural mineral waters (still), effervescent natural mineral waters (sparkling) and aromatized waters with fruit-flavors (still or sparkling) are an emerging market. In this work, the capability of a potentiometric electronic tongue, comprised with lipid polymeric membranes, to quantitatively estimate routinely quality physicochemical parameters (pH and conductivity) as well as to qualitatively classify water samples according to the type of water was evaluated. The study showed that a linear discriminant model, based on 21 sensors selected by the simulated annealing algorithm, could correctly classify 100 % of the water samples (leave-one out cross-validation). This potential was further demonstrated by applying a repeated K-fold cross-validation (guaranteeing that at least 15 % of independent samples were only used for internal-validation) for which 96 % of correct classifications were attained. The satisfactory recognition performance of the E-tongue could be attributed to the pH, conductivity, sugars and organic acids contents of the studied waters, which turned out in significant differences of sweetness perception indexes and total acid flavor. Moreover, the E-tongue combined with multivariate linear regression models, based on sub-sets of sensors selected by the simulated annealing algorithm, could accurately estimate waters pH (25 sensors: R 2 equal to 0.99 and 0.97 for leave-one-out or repeated K-folds cross-validation) and conductivity (23 sensors: R 2 equal to 0.997 and 0.99 for leave-one-out or repeated K-folds cross-validation). So, the overall satisfactory results achieved, allow envisaging a potential future application of electronic tongue devices for bottled water analysis and classification.

Electroquímica de sustancias de interés en los sistemas agroalimentario y de sanidad animal. Desarrollo de biosensores electroquímicos. Aplicaciones analíticas.Electrochemistry of substances of interest in agroalimentary and animal health systems. Development of electrochemical biosensors. Analytical applications.

Se estudiarán los mecanismos de reacción electroquímica de las micotoxinas (metabolitos tóxicos generados por hongos) citrinina (CIT), patulina (PAT) y moniliformina (MON), de los antioxidantes naturales alfa, beta, gama y delta tocoferoles, de los flavonoides fisetina (FIS), morina (MOR), luteolina (LUT), rutina (RUT), buteina (BUT), naringenina (NAR) y miricetina (MIR) y de las hormonas esteroides estradiol (EDIOL), estrona (EONA) y estriol (ETRIOL). Por otra parte, se implementarán técnicas electroanalíticas para la detección y cuantificación de estos sustratos en muestras de matrices naturales que los contengan. Se realizará el diseño y caracterización de biosensores enzimáticos a partir de peroxidasas y/o fosfatasa alcalina para la determinación de la micotoxina CIT y de los flavonoides y, por otro, de inmunosensores para las micotoxinas ocratoxina A (OTA) y PAT y hormonas. Para el anclaje de enzimas y/o anticuerpos, se estudiarán las propiedades de electrodos modificados por monocapas autoensambladas, nanotubos de carbono y partículas magnéticas. Se usarán las técnicas de voltamperometría cíclica, de onda cuadrada y de redisolución con acumulación adsortiva, espectroscopías de impedancia electroquímica, electrólisis a potencial controlado, uv-vis e IR, microbalanza de cristal de cuarzo y microscopías de alta resolución (SEM, TEM, AFM). La importancia de este proyecto apunta a la obtención de nuevos datos electroquímicos de los sustratos indicados y conocimientos relacionados con la aplicación de electrodos modificados en la preparación de biosensores y en el desarrollo de técnicas alternativas para la determinación de los analitos mencionados precedentemente. Electrochemical reaction mechanisms of mycotoxins (toxic metabolites generated by fungi) citrinin (CIT), Patulin (PAT) and moniliformin (MON), natural antioxidants alpha, beta, gamma and delta tocopherols, flavonoids fisetin (FIS), morin (MOR), luteolin (LUT), rutin (RUT), butein (BUT), naringenin (NAR), miricetin (MIR) and steroid hormones estradiol (EDIOL), estrone (EONA) and estriole (ETRIOL) will be explored. On the other hand, electroanalytical techniques for the detection and quantification of these substrates in samples of natural matrices will be implemented. The design and characterization of enzymatic biosensors from peroxidases and/or from alkaline phosphatase for the determination of CIT and flavonoids, and also of inmunosensors for ochratoxin A (OTA) and PAT and hormones will be performed. For the anchor of enzymes and/or antibody, properties of electrodes modified by self assembled monolayers, carbon nanotubes and magnetic particles will be explored. Cyclic, square wave and adsorptive stripping voltammetries, electrochemical impedance spectroscopy, controlled potential electrolysis, uv-vis and IR, quartz crystal microbalance and high-resolution microcopies (SEM, TEM, AFM) will be used. The importance of this project is aimed at obtaining new electrochemical data for the indicated substrates and knowledge on the application of modified electrodes in preparation of biosensors and in the development of alternative techniques for the determination of the above-mentioned analytes.

mRNA metabolism: nonsense mediated mRNA decay as a tool for gene therapy and the role of human DIS3L2 in transcript degradation

Tese de mestrado em Biologia Humana e Ambiente, apresentada à Universidade de Lisboa, através da Faculdade de Ciências, 2015