477 research outputs found

    Celiac Immunogenic Potential of α-Gliadin Epitope Variants from Triticum and Aegilops Species

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    The high global demand of wheat and its subsequent consumption arise from the physicochemical properties of bread dough and its contribution to the protein intake in the human diet. Gluten is the main structural complex of wheat proteins and subjects affected by celiac disease (CD) cannot tolerate gluten protein. Within gluten proteins, α-gliadins constitute the most immunogenic fraction since they contain the main T-cell stimulating epitopes (DQ2.5-glia-α1, DQ2.5-glia-α2, and DQ2.5-glia-α3). In this work, the celiac immunotoxic potential of α-gliadins was studied within Triticeae: diploid, tetraploid, and hexaploid species. The abundance and immunostimulatory capacity of CD canonical epitopes and variants (with one or two mismatches) in all α-gliadin sequences were determined. The results showed that the canonical epitopes DQ2.5-glia-α1 and DQ2.5-glia-α3 were more frequent than DQ2.5-glia-α2. A higher abundance of canonical DQ2.5-glia-α1 epitope was found to be associated with genomes of the BBAADD, AA, and DD types; however, the abundance of DQ2.5-glia-α3 epitope variants was very high in BBAADD and BBAA wheat despite their low abundance in the canonical epitope. The most abundant substitution was that of proline to serine, which was disposed mainly on the three canonical DQ2.5 domains on position 8. Interestingly, our results demonstrated that the natural introduction of Q to H at any position eliminates the toxicity of the three T-cell epitopes in the α-gliadins. The results provided a rational approach for the introduction of natural amino acid substitutions to eliminate the toxicity of three T-cell epitopes, while maintaining the technological properties of commercial wheats

    Simultaneous determination of traces of PT, PD, OS, IR, RH, AG and AU by using magnetic nanoparticles solid phase extraction coupled with ICP OES

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    The direct analysis of these target analytes is very limited being essential sample pre-treatment techniques and the use of very sensitive instrumental techniques to carry out determinations. The inductively coupled plasma optical emission spectrometry shows a poor sensitivity because the concentration of some elements in environmental samples is below the detection limit of ICP OES. To solve this problem, preconcentration separation procedures have been proposed, minimizing the spectral and matrix interferences. Thus, enrichment is a very important issue for achievement of low detection limits [1-4]. In this study, a chelating resin 1,5 bis (di 2 pyridil) methylene thiocarbonohydrazide bonded to iron oxide magnetic nanoparticles (DPTH-MNPs) were synthesized. These magnetic nanoparticles were employed as a solid phase extraction (SPE) adsorbent for the separation and concentration of trace amounts of 7 elements (Au, Ag, Pd, Pt, Ir, Rh and Os) from environmental water samples. The main aim of this work was to develop a precise and accurate method for the simultaneous determination of the maximum possible number of elements by using this new absorbent and a multimode sample introduction system (MSIS). The MSIS acts as a system for the generation, separation and introduction of chemical vapours (CVG) and also as an introduction system for sample aerosols, in a simultaneous form, into an inductively coupled plasma-optical emission spectrometer. The on-line SPE-CVG-ICP-OES system developed was applied in the determination of the aforementioned metals in natural water samples (sea water, estuarine, lake and river water), with the least demanding and simple sample preparation procedure. The developed method was validated by analysing natural water certified reference materials (TMDA 54.4 fortified lake waters and SRM 1643e, trace elements in water; and National Institute of Standards and Technology (NIST), NIST-2557 autocatalyst). Sea water, tap water and well water samples collected from Malaga (Spain) were also analysed. The procedure has been demonstrated to be fast, easy, automatic, selective and economical, and the sensitivity was good. The main advantage of DPTH-MNPs is its very good stability and resistance because chemisorption of chelating molecules on the surface of solid supports provides immobility, mechanical stability and insolubility. The precision (RSD), accuracy (by standard addition or recovery) and limit of detection (LOD) were used to evaluate the characteristics of the procedure. Furthermore, the proposed method was applied in the simultaneous determination of the 7 elements mentioned above with a sample throughput of about 13 h-1, thereby, reducing the time of analysis and the volume of reagents and sample required. References [1] M. Tuzen, M. Soylak, D. Citak, H.S. Ferreira, M.G.A. Korn, M.A. Bezerra, A pre-concentration system for determination of copper and nickel in water and food samples employing flame atomic absorption spectrometry, Journal of Haz-ardous Materials 162 (2009) 1041–1045. [2] Y. Cui, X. Chang, Y. Zhai, X. Zhu, H. Zheng, N. Lian, ICP-AES determination of trace elements after preconcentrated with p-dimethylaminobenzaldehyde-modified nanometer SiO2 from sample solution, Microchem. J. 83 (2006) 35–41. [3] P. Liang, B. Hu, Z. Jiang, Y. Qin, T. Peng, Nanometer-sized titanium dioxide micro-column on-line preconcentration of La, Y, Yb, Eu, Dy and their determination by inductively coupled plasma atomic emission spectrometry, J. Anal. Atom. Spectrom. 16 (2001) 863–866. [4] B. Feist, B. Mikula, K. Pytlakowska, B. Puzio, F. Buhl, Determination of heavy metals by ICP-OES and F-AAS after preconcentration with 2,2-bipyridyl and erythrosine, J. Hazard. Mater. 152 (2008) 1122–1129.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

    Sequential determination of traces of As, Sb and hg by on-line magnetic solid phase extraction coupled with Hr-Cs-Cvg-Gfaas

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    A green and rapid method was developed for the simultaneous separation/preconcentration and sequential monitoring pf arsenic, antimony and mercury by flow injection magnetic solid phase extraction coupled with on-line chemical vapor generation and determination by high resolution continuum source graphite furnace atomic absorption spectrometry. The system is based on chelating/cationic retention of the analytes onto a magnet based reactor designed to contain functionalized magnetic nanoparticles (MNPs). The MNP score allows overcoming the back-pressure problems that usually happen in SPME methods with NPs thanks to the possibility of inmobilizing the MNPs by applying an external magnetic field. Several chemical and flow variables were considered as factors in the optimization process using central composite designs. With the optimized procedure the detection limits obtained were 0.2, 0.003 and 0.4 µg/L for As, Sb and Hg respectively. For the quality control of the analytical performance and the validation of the developed method the analysis of two certified samples TM 24.3 and TMDA 54.4 Fortified Lake Waters was addressed. The results showed good agreement with the certified values.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

    Stimulatory Response of Celiac Disease Peripheral Blood Mononuclear Cells Induced by RNAi Wheat Lines Differing in Grain Protein Composition

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    Wheat gluten proteins are responsible for the bread-making properties of the dough but also for triggering important gastrointestinal disorders. Celiac disease (CD) affects approximately 1% of the population in Western countries. The only treatment available is the strict avoidance of gluten in the diet. Interference RNA (RNAi) is an excellent approach for the down-regulation of genes coding for immunogenic proteins related to celiac disease, providing an alternative for the development of cereals suitable for CD patients. In the present work, we report a comparative study of the stimulatory capacity of seven low-gluten RNAi lines differing in grain gluten and non-gluten protein composition, relevant for CD and other gluten pathologies. Peripheral blood mononuclear cells (PBMCs) of 35 patients with active CD were included in this study to assess the stimulatory response induced by protein extracts from the RNAi lines. Analysis of the proliferative response and interferon-gamma (INF-γ) release of PBMCs demonstrated impaired stimulation in response to all RNAi lines. The lower response was provided by lines with a very low content of α- and γ-gliadins, and low or almost devoid of DQ2.5 and p31–43 α-gliadin epitopes. The non-gluten protein seems not to play a key role in PBMC stimulation.Spanish Ministry of Economy, Industry and competitiveness AGL2016-80566-PEuropean Regional Development Fund (FEDER

    Cold vapour generation electrothermal atomic absorption spectrometry and solid phase extraction based on a new nanosorbent for sensitive HG determination in environmental samples (sea water and river water)

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    Reunión bianual del Grupo Regional Andaluz de la Sociedad Española de Química AnalíticaMercury is not an essential element for plant or animal life and it is a potential environmental toxic because of its tendency to form covalent bonds with organic molecules and the high stability of the Hg-C bond. Reports estimate a total mercury concentration in natural waters ranging from 0.2 to 100 ng L-1. Due to this fact, highly sensitive methods are required for direct determination of such extremely low levels. In this work, a rapid and simple method was developed for separation and preconcentration of mercury by flow injection solid phase extraction coupled with on-line chemical vapour generation electrothermal atomic absorption spectrometry. The system is based on chelating retention of the analyte onto the mini column filled with a mesoporous silica functionalized with 1,5 bis (di-2-pyridyl) methylene thiocarbohydrazide. The main aim of this work was to develop a precise and accurate method for the determination of the Hg. Under the optima conditions and 120 s preconcentration time, the detection limit obtained was 0.009 μg L-1, with RSDs 3.7 % for 0.2 μg L-1, 4.8 % for 1 μg L-1 and enrichment factor 4, Furthermore, the method proposed has permitted the determination of Hg with a reduction in the analysis time, the sample throughput was about 18 h-1, low consumption of reagents and sample volume. The method was applied to the determination of Hg in sea water and river water. For the quality control of the analytical performance and the validation of the newly developed method, the analysis of two certified samples, TMDA 54.4 Fortified Lake, and LGC6187 River sediment was addressed. The results showed good agreement with the certified values.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

    Determination of Pb using F3eO4 GO join to DPTH for ferrofluid based dispersive solid phase extraction

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    In this work has been described a green and rapid method the synthesis of Fe3O4@GO nanospheres via chemical covalent bonding method. The Fe3O4@GO DPTH was applied to ferrofluid based dispersive solid phase extraction of lead as a model analyte using an ionic liquid carrier. The ferro fluid allows the rapid extraction of lead ions using a low amount of sorbent material. Besides, the magnetic separation greatly improved the separation rate. The presented method is highly time saving due to the high dispersion of the sorbent in the aqueous phase and also there is no need to shake the sample solution. The other benefits of the proposed methods are simplicity of operation, low cost, high sorption capacity, high recovery and high preconcentration. In order to optimize the method the following parameters were studied: sample solution pH, concentration of DPTH, extraction time, amount of sorbent, desorption conditions, influence of ionic strength, and tolerance of potentially interfering ions. The sample or standard solution containing Pb(II), DPTH (0.05% ethanol w/v), NaCl (0.5 %, w/v) and buffer (pH = 5.6) was poured into high volume. Then 240 µL of ferrofluid was injected rapidly into the sample solution through a syringe. Thereupon, a dark cloudy suspension was formed, ferrofluid was dispersed thoroughly in solution and the complex of Pb-DPTH was extracted in a few seconds. Subsequently, a strong magnet was placed at the bottom of the tube to let the extractant settle. After about 3 min, the solution became clear and the supernatant was discarded simply by decanting it. Afterwards, the magnet was removed and 1 mL of nitric acid (2.0 mol L−1) was introduced to the vial to desorb the Pb by sonication. Finally, the sorbent was separated by positioning the magnet to the outside of the tube and the concentration of Pb in acidic aqueous phase was determined by ETAAS.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

    Propiedades funcionales de proteínas de soja parcialmente hidrolizadas empleando la peptidasa de Asclepias fruticosa

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    La harina de soja es el subproducto más relevante en el procesamiento de esta leguminosa y contiene un alto contenido de proteínas. La hidrólisis controlada de dichas proteínas puede generar ingredientes de alto valor añadido, con mejores características nutricionales y propiedades funcionales. La preparación proteolítica denominada Af, obtenida por centrifugación del látex de Asclepias fruticosa (Asclepiadaceae) recolectado en buffer cítrico-citrato, posee una única peptidasa cisteínica de PM 23,7 kDa y pI>9,3 denominada asclepaína f, la cual ha sido clonada y expresada en Pichia pastoris. La preparación Af posee una actividad proteolítica de 1,1 Ucas/ml, alta estabilidad térmica a 37° y 45°C, un rango de pH óptimo de actividad entre 7,5 y 10,5 y ha demostrado capacidad para hidrolizar las proteínas de soja, degradando fuertemente la fracción 7S y en menor medida las globulinas 11S. El objetivo del trabajo fue evaluar las propiedades funcionales (solubilidad, capacidad de retención de agua y aceite, y espumado) de un hidrolizado de proteínas de soja obtenido con la preparación Af. Se utilizó una dispersión de concentrado de soja (conteniendo 62,1 % de proteínas determinadas por Kjeldhal) al 8% p/v y una cantidad de extracto enzimático equivalente a 8,9 Ucas por gramo de proteína, con el agregado de cisteína 20 mM. La hidrólisis se llevó a cabo a diferentes valores de pH (8, 9 y 10) durante 180 minutos a 45ºC. La actividad enzimática se detuvo por calentamiento en microondas, las suspensiones fueron centrifugadas y los pellets secados en estufa. Se realizó un blanco de reacción en las mismas condiciones, reemplazando la preparación enzimática por igual volumen de agua. Posteriormente, se determinó la concentración de proteínas solubles mediante el método de Bradford, la capacidad de retención de agua a diferentes temperaturas (5, 20 y 45°C), la retención de aceite de soja y girasol, y la formación de espuma. La capacidad de retención de agua se ensayó mezclando 1 gr de muestra con 10 ml de agua destilada a diferentes temperaturas, centrifugando las suspensiones y determinando el peso de las muestras húmedas; del mismo modo se determinó la capacidad de retención de aceite. Para ensayar la capacidad espumante se prepararon suspensiones al 1% p/v de los hidrolizados, se agitaron durante 1 minuto a temperatura ambiente y posteriormente se midió el volumen de espuma y de suspensión. El mayor grado de hidrólisis de las proteínas de soja con la preparación Af se obtuvo a pH 10 y fue del 7%. A través de dicho proceso de hidrólisis se logró incrementar la concentración de proteínas solubles un 60%, la capacidad de retención de agua a 20°C se incrementó un 71% y a 45°C un 134%, mientras que la capacidad de retención de aceite de soja a 20°C aumentó un 92% y la de girasol 73,5%. En tanto que la capacidad espumante casi no fue afectada, se detectó solo un ligero aumento (3%). Empleando la preparación Af se logró obtener un hidrolizado de proteínas de soja con propiedades funcionales particulares que podrá emplearse en el desarrollo de alimentosInstituto Multidisciplinario de Biología Celula

    High-Throughput System for the Early Quantification of Major Architectural Traits in Olive Breeding Trials Using UAV Images and OBIA Techniques

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    The need for the olive farm modernization have encouraged the research of more efficient crop management strategies through cross-breeding programs to release new olive cultivars more suitable for mechanization and use in intensive orchards, with high quality production and resistance to biotic and abiotic stresses. The advancement of breeding programs are hampered by the lack of efficient phenotyping methods to quickly and accurately acquire crop traits such as morphological attributes (tree vigor and vegetative growth habits), which are key to identify desirable genotypes as early as possible. In this context, an UAV-based high-throughput system for olive breeding program applications was developed to extract tree traits in large-scale phenotyping studies under field conditions. The system consisted of UAV-flight configurations, in terms of flight altitude and image overlaps, and a novel, automatic, and accurate object-based image analysis (OBIA) algorithm based on point clouds, which was evaluated in two experimental trials in the framework of a table olive breeding program, with the aim to determine the earliest date for suitable quantifying of tree architectural traits. Two training systems (intensive and hedgerow) were evaluated at two very early stages of tree growth: 15 and 27 months after planting. Digital Terrain Models (DTMs) were automatically and accurately generated by the algorithm as well as every olive tree identified, independently of the training system and tree age. The architectural traits, specially tree height and crown area, were estimated with high accuracy in the second flight campaign, i.e. 27 months after planting. Differences in the quality of 3D crown reconstruction were found for the growth patterns derived from each training system. These key phenotyping traits could be used in several olive breeding programs, as well as to address some agronomical goals. In addition, this system is cost and time optimized, so that requested architectural traits could be provided in the same day as UAV flights. This high-throughput system may solve the actual bottleneck of plant phenotyping of "linking genotype and phenotype," considered a major challenge for crop research in the 21st century, and bring forward the crucial time of decision making for breeders

    Danthron, an Anthraquinone Isolated from a Marine Fungus, Is a New Inhibitor of Angiogenesis Exhibiting Interesting Antitumor and Antioxidant Properties

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    The role played by a sustained angiogenesis in cancer and other diseases stimulates the interest in the search for new antiangiogenic drugs. In this manuscript, we provide evidence that 1,8- dihydroxy-9,10-anthraquinone (danthron), isolated from the fermentation broth of the marine fungus Chromolaenicola sp. (HL-114-33-R04), is a new inhibitor of angiogenesis. The results obtained with the in vivo CAM assay indicate that danthron is a potent antiangiogenic compound. In vitro studies with human umbilical endothelial cells (HUVEC) reveal that this anthraquinone inhibits certain key functions of activated endothelial cells, including proliferation, proteolytic and invasive capabilities and tube formation. In vitro studies with human breast carcinoma MDA-MB231 and fibrosarcoma HT1080 cell lines suggest a moderate antitumor and antimetastatic activity of this compound. Antioxidant properties of danthron are evidenced by the observation that it reduces the intracellular reactive oxygen species production and increases the amount of intracellular sulfhydryl groups in endothelial and tumor cells. These results support a putative role of danthron as a new antiangiogenic drug with potential application in the treatment and angioprevention of cancer and other angiogenesis-dependent diseases.Partial funding for open access charge: Universidad de Málag
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