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Teixeira A.S.,ICTAN CSIC | Teixeira A.S.,CONICET | Molina-Garcia A.D.,ICTAN CSIC
Refrigeration Science and Technology | Year: 2015

Biological samples are successfully preserved for indefinite periods after vitrification, providing ice formation is avoided. Glass is difficult to characterize, especially at the extremely low temperatures required for biological tissues vitrification. Glass transition is traditionally observed by differential scanning calorimetry (DSC), in spite of its reduced sensitivity and lack of spatial information. In this work, a novel procedure, based on low-temperature scanning electron microscopy (cryo-SEM), is explored to locate vitrified areas and ice crystals in tissues and cells. Cryo-SEM observation of biological samples requires an etching phase to create contrast: a temperature rise, allowing ice partial sublimation. Glassy water, differently from crystallized water (ice), has a neglectable sublimation rate. Consequently, while the dark image of sublimated crystals and the resulting structural details can be observed in ice-containing samples, vitrified tissues show a smooth landscape, without dark areas or visible structure elements.


Diaz-Rubio M.E.,Institute Ciencia Y Tecnologia Of Alimentos Y Nutricion Ictan | Serrano J.,Institute Ciencia Y Tecnologia Of Alimentos Y Nutricion Ictan | Borderias J.,ICTAN CSIC | Saura-Calixto F.,Institute Ciencia Y Tecnologia Of Alimentos Y Nutricion Ictan
Journal of Aquatic Food Product Technology | Year: 2011

The technological effect of antioxidant dietary fiber (AODF) from Fucus vesiculosus added to minced horse mackerel (Trachurus trachurus) during frozen storage was tested. Fish mince (FM) samples supplemented with 1 and 2% AODF were compared to AODF-free control. Dietary analysis, antioxidant capacity, total polyphenols content, ferric-reducing antioxidant power (FRAP assay), and free radical scavenging activity (by ABTS assay), water binding capacity, water holding ability, cooking yield, and sensory analyses were carried out just after freezing and during frozen storage. Samples with 1 and 2% of AODF added had lower lipid oxidation than those without AODF, and the total drip (thaw + cooking) was reduced after 3 months of frozen storage. Fish samples supplemented with 1% AODF did not have a different flavor from the control. Samples supplemented with 2% AODF had a different flavor from the control, but this did not impair palatability. Copyright © Taylor & Francis Group, LLC.


Olivares M.,CSIC - Institute of Agricultural Chemistry and Food Technology | Neef A.,CSIC - Institute of Agricultural Chemistry and Food Technology | Castillejo G.,Hospital Universitario Sant Joan Of Reus | De Palma G.,CSIC - Institute of Agricultural Chemistry and Food Technology | And 9 more authors.
Gut | Year: 2014

Objective Intestinal dysbiosis has been associated with coeliac disease (CD), but whether the alterations are cause or consequence of the disease is unknown. This study investigated whether the human leukocyte antigen (HLA)-DQ2 genotype is an independent factor influencing the early gut microbiota composition of healthy infants at family risk of CD. Design As part of a larger prospective study, a subset (n=22) of exclusively breastfed and vaginally delivered infants with either high genetic risk (HLA-DQ2 carriers) or low genetic risk (non-HLA-DQ2/8 carriers) of developing CD were selected from a cohort of healthy infants with at least one first-degree relative with CD. Infant faecal microbiota was analysed by 16S rRNA gene pyrosequencing and real time quantitative PCR. Results Infants with a high genetic risk had significantly higher proportions of Firmicutes and Proteobacteria and lower proportions of Actinobacteria compared with low-risk infants. At genus level, high-risk infants had significantly less Bifidobacterium and unclassified Bifidobacteriaceae proportions and more Corynebacterium, Gemella, Clostridium sensu stricto, unclassified Clostridiaceae, unclassified Enterobacteriaceae and Raoultella proportions. Quantitative real time PCR also revealed lower numbers of Bifidobacterium species in infants with high genetic risk than in those with low genetic risk. In high-risk infants negative correlations were identified between Bifidobacterium species and several genera of Proteobacteria (Escherichia/Shigella) and Firmicutes (Clostridium). Conclusions The genotype of infants at family risk of developing CD, carrying the HLA-DQ2 haplotypes, influences the early gut microbiota composition. This finding suggests that a specific disease-biased host genotype may also select for the first gut colonisers and could contribute to determining disease risk. © 2014 BMJ Publishing Group Ltd & British Society of Gastroenterology.


Pereira-Caro G.,UK Institute of Food Research | Pereira-Caro G.,IFAPA Centro del Llano | Mateos R.,UK Institute of Food Research | Traka M.H.,UK Institute of Food Research | And 5 more authors.
Food Chemistry | Year: 2013

The anticarcinogenic activity of hydroxytyrosyl ethyl ether (HTy-Et) compared to its precursor hydroxytyrosol (HTy) has been studied in human Caco-2 colon adenocarcinoma cells. 451 and 977 genes were differentially expressed in Caco-2 cells exposed to HTy or HTy-Et for 24 h, respectively, compared with untreated cells (P < 0.005; FDR = 0), using Affymetrix microarrays. Results showed that both HTy and HTy-Et inhibited cell proliferation and arrested the cell cycle by up-regulating p21 and CCNG2 and down-regulating CCNB1 protein expression. HTy and HTy-Et also altered the transcription of specific genes involved in apoptosis, as suggested by the up-regulation of BNIP3, BNIP3L, PDCD4 and ATF3 and the activation of caspase-3. Moreover, these polyphenols up-regulated xenobiotic metabolizing enzymes UGT1A10 and CYP1A1, enhancing carcinogen detoxification. In conclusion, these results highlight that HTy and its derivative HTy-Et modulate molecular mechanisms involved in colon cancer, with HTy-Et being more effective than HTy. © 2012 Elsevier Ltd. All rights reserved.


Mateos R.,UK Institute of Food Research | Pereira-Caro G.,UK Institute of Food Research | Pereira-Caro G.,IFAPA Centro del Llano | Bacon J.R.,UK Institute of Food Research | And 4 more authors.
Journal of Agricultural and Food Chemistry | Year: 2013

The anticancer activity of hydroxytyrosyl acetate (HTy-Ac) has been studied in human colon adenocarcinoma cells. Gene expression of proteins involved in cell cycle (p21, p53, cyclin B1, and cyclin G2) and programmed cell death (BNIP3, BNIP3L, PDCD4, and ATF3), as well as phase I and phase II detoxifying enzymes CYPA1 and UGT1A10, were evaluated by reverse transcription polymerase chain reaction after 24 h of exposure of Caco-2/TC7 cells to 5, 10, and 50 μM of HTy-Ac. The results show that HTy-Ac inhibited cell proliferation and arrested cell cycle by enhancing p21 and CCNG2 and lowering CCNB1 protein expression. HTy-Ac also affected the transcription of genes involved in apoptosis up-regulating of BNIP3, BNIP3L, PDCD4, and ATF3 and activating caspase-3. In addition, HTy-Ac also up-regulated xenobiotic metabolizing enzymes CYP1A1 and UGT1A10, thus enhancing carcinogen detoxification. In conclusion, these results highlight that HTy-Ac has the potential to modulate biomarkers involved in colon cancer. © 2013 American Chemical Society.


Olivares M.,CSIC - Institute of Agricultural Chemistry and Food Technology | Neef A.,CSIC - Institute of Agricultural Chemistry and Food Technology | Castillejo G.,Hospital Universitario Sant Joan Of Reus | De Palma G.,CSIC - Institute of Agricultural Chemistry and Food Technology | And 10 more authors.
Gut | Year: 2015

Objective: Intestinal dysbiosis has been associated with coeliac disease (CD), but whether the alterations are cause or consequence of the disease is unknown. This study investigated whether the human leukocyte antigen (HLA)-DQ2 genotype is an independent factor influencing the early gut microbiota composition of healthy infants at family risk of CD. Design: As part of a larger prospective study, a subset (n=22) of exclusively breastfed and vaginally delivered infants with either high genetic risk (HLA-DQ2 carriers) or low genetic risk (non-HLA-DQ2/8 carriers) of developing CD were selected from a cohort of healthy infants with at least one first-degree relative with CD. Infant faecal microbiota was analysed by 16S rRNA gene pyrosequencing and real time quantitative PCR. Results: Infants with a high genetic risk had significantly higher proportions of Firmicutes and Proteobacteria and lower proportions of Actinobacteria compared with low-risk infants. At genus level, high-risk infants had significantly less Bifidobacterium and unclassified Bifidobacteriaceae proportions and more Corynebacterium, Gemella, Clostridium sensu stricto, unclassified Clostridiaceae, unclassified Enterobacteriaceae and Raoultella proportions. Quantitative real time PCR also revealed lower numbers of Bifidobacterium species in infants with high genetic risk than in those with low genetic risk. In high-risk infants negative correlations were identified between Bifidobacterium species and several genera of Proteobacteria (Escherichia/Shigella) and Firmicutes (Clostridium). Conclusions: The genotype of infants at family risk of developing CD, carrying the HLA-DQ2 haplotypes, influences the early gut microbiota composition. This finding suggests that a specific disease-biased host genotype may also select for the first gut colonisers and could contribute to determining disease risk.


de Palma G.,IATA CSIC | Capilla A.,CIBER ISCIII | Nova E.,ICTAN CSIC | Castillejo G.,Hospital Universitario Sant Joan Of Reus | And 13 more authors.
PLoS ONE | Year: 2012

Interactions between environmental factors and predisposing genes could be involved in the development of coeliac disease (CD). This study has assessed whether milk-feeding type and HLA-genotype influence the intestinal microbiota composition of infants with a family history of CD. The study included 164 healthy newborns, with at least one first-degree relative with CD, classified according to their HLA-DQ genotype by PCR-SSP DQB1 and DQA1 typing. Faecal microbiota was analysed by quantitative PCR at 7 days, and at 1 and 4 months of age. Significant interactions between milk-feeding type and HLA-DQ genotype on bacterial numbers were not detected by applying a linear mixed-model analysis for repeated measures. In the whole population, breast-feeding promoted colonization of C. leptum group, B. longum and B. breve, while formula-feeding promoted that of Bacteroides fragilis group, C. coccoides-E. rectale group, E. coli and B. lactis. Moreover, increased numbers of B. fragilis group and Staphylococcus spp., and reduced numbers of Bifidobacterium spp. and B. longum were detected in infants with increased genetic risk of developing CD. Analyses within subgroups of either breast-fed or formula-fed infants indicated that in both cases increased risk of CD was associated with lower numbers of B. longum and/or Bifidobacterium spp. In addition, in breast-fed infants the increased genetic risk of developing CD was associated with increased C. leptum group numbers, while in formula-fed infants it was associated with increased Staphylococcus and B. fragilis group numbers. Overall, milk-feeding type in conjunction with HLA-DQ genotype play a role in establishing infants' gut microbiota; moreover, breast-feeding reduced the genotype-related differences in microbiota composition, which could partly explain the protective role attributed to breast milk in this disorder. © 2012 De Palma et al.


Magrone T.,University of Bari | de Heredia F.P.,ICTAN CSIC | de Heredia F.P.,Liverpool John Moores University | Jirillo E.,University of Bari | And 4 more authors.
Canadian Journal of Physiology and Pharmacology | Year: 2013

In Western societies, the incidence of diet-related diseases is progressively increasing due to greater availability of hypercaloric food and a sedentary lifestyle. Obesity, diabetes, atherosclerosis, and neurodegeneration are major diet-related pathologies that share a common pathogenic denominator of low-grade inflammation. Functional foods and nutraceuticals may represent a novel therapeutic approach to prevent or attenuate diet-related disease in view of their ability to exert antiinflammatory responses. In particular, activation of intestinal T regulatory cells and homeostatic regulation of the gut microbiota have the potential to reduce low-grade inflammation in diet-related diseases. In this review, clinical applications of polyphenol-rich functional foods and nutraceuticals in postprandial inflammation, obesity, and ageing will be discussed. We have placed special emphasis on polyphenols since they are broadly distributed in plants.


Saura-Calixto F.,ICTAN CSIC | Perez-Jimenez J.,ICTAN CSIC | Perez-Jimenez J.,French National Institute for Agricultural Research | Tourino S.,CSIC - Institute of Advanced Chemistry of Catalonia | And 6 more authors.
Molecular Nutrition and Food Research | Year: 2010

Proanthocyanidins (PAs) or condensed tannins, a major group of dietary polyphenols, are oligomers and polymers of flavan-3-ol and flavan-3, 4-diols widely distributed in plant foods. Most literature data on PAs' metabolic fate deal with PAs that can be extracted from the food matrix by aqueous-organic solvents (extractable proanthocyanidins). However, there are no data on colonic fermentation of non-extractable proanthocyanidins (NEPAs), which arrive almost intact to the colon, mostly associated to dietary fibre (DF). The aim of the present work was to examine colonic fermentation of NEPAs associated with DF, using a model of in vitro small intestine digestion and colonic fermentation. Two NEPA-rich materials obtained from carob pod (Ceratonia siliqua L. proanthocyanidin) and red grapes (grape antioxidant dietary fibre) were used as test samples. The colonic fermentation of these two products released hydroxyphenylacetic acid, hydroxyphenylvaleric acid and two isomers of hydro-xyphenylpropionic acid, detected by HPLC-ESI-MS/MS. Differences between the two products indicate that DF may enhance the yield of metabolites. In addition, the main NEPA metabolite in human plasma was 3,4-dihydroxyphenyl acetic acid. The presence in human plasma ofthe same metabolites as were detected after in vitro colonic fermentation ofNEPAs suggests that dietary NEPAs would undergo colonic fermentation releasing absorbable metabolites with potential healthy effects. © 2010 WILEY-VCH Verlag GmbH & Co.


Queguineur B.,National University of Ireland | Goya L.,ICTAN CSIC | Ramos S.,ICTAN CSIC | Martin M.A.,ICTAN CSIC | And 2 more authors.
Food and Chemical Toxicology | Year: 2012

Phloroglucinol is an ubiquitous secondary metabolite encountered in a free state or polymerised as phlorotannins in brown macroalgae, and present in higher plants. FRAP and TEAC assays measured the antioxidant properties of phloroglucinol in non-biological conditions. Additionally, the biological effects of phloroglucinol (4-400 μM) were scrutinised using cellular oxidative stress markers, such as the generation of ROS, antioxidant defences (concentration of GSH and activities of GPx, GR and GST), and levels of MDA as a marker for lipid peroxidation. The direct effect was assessed immediately after an incubation period, whereas for the protective effect, the incubation period was followed by 3-h treatment with the pro-oxidant t-BOOH. The results indicated that despite having a higher radical scavenging capacity than Trolox after 30. min, phloroglucinol was not a suitable antioxidant standard for phlorotannins. Regarding the biological effects, phloroglucinol had no impact on cell viability, reduced levels of ROS and increased antioxidant defences in the direct treatment for most concentrations. The results of the protective effect were mitigated as phloroglucinol failed to protect from ROS generation but evoked a significant recovery of the stress-altered cellular antioxidant defences to restful conditions. Additionally, MDA levels were greatly reduced, preventing a radical chain oxidation. © 2012 Elsevier Ltd.

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