José M. Gallardo

Spanish National Research Council, Madrid, Madrid, Spain

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Publications (113)248.32 Total impact

  • Ignacio Ortea, José M. Gallardo
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    ABSTRACT: Three factors defining the traceability of a food product are production method (wild or farmed), geographical origin and biological species, which have to be checked and guaranteed, not only in order to avoid mislabelling and commercial fraud, but also to address food safety issues and to comply with legal regulations. The aim of this study was to determine whether these three factors could be differentiated in shrimps using stable isotope ratio analysis of carbon and nitrogen and/or multi-element composition. Different multivariate statistics methods were applied to different data subsets in order to evaluate their performance in terms of classification or predictive ability. Although the success rates varied depending on the dataset used, the combination of both techniques allowed the correct classification of 100% of the samples according to their actual origin and method of production, and 93.5% according to biological species. Even though further studies including a larger number of samples in each group are needed in order to validate these findings, we can conclude that these methodologies should be considered for studies regarding seafood product authenticity.
    Food Chemistry 03/2015; 170:145–153. DOI:10.1016/j.foodchem.2014.08.049 · 3.26 Impact Factor
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    ABSTRACT: The present research draws a map of the characteristic carbonylation of proteins in rats fed high caloric diets with the aim of providing a new insight of the pathogenesis of metabolic diseases derived from the high consumption of fat and refined carbohydrates. Protein carbonylation was analyzed in plasma, liver and skeletal muscle of Sprague-Dawley rats fed a high-fat high-sucrose (HFHS) diet by a proteomics approach based on carbonyl-specific fluorescence-labelling, gel electrophoresis and mass spectrometry. Oxidized proteins along with specific sites of oxidative damage were identified and discussed to illustrate the consequences of protein oxidation. The results indicated that long-term HFHS consumption increased protein oxidation in plasma and liver, meanwhile protein carbonyls from skeletal muscle did not change. The increment of carbonylation by HFHS diet was singularly selective on specific target proteins: albumin from plasma and liver, and hepatic proteins such as mitochondrial carbamoyl-phosphate synthase (ammonia), mitochondrial aldehyde dehydrogenase, argininosuccinate synthetase, regucalcin, mitochondrial ATP synthase subunit beta, actin cytoplasmic 1, mitochondrial glutamate dehydrogenase 1. The possible consequences that these specific protein carbonylations have on the excessive weight gain, insulin resistance and nonalcoholic fatty liver disease resulting from HFHS diet consumption have been discussed.
    The Journal of Nutritional Biochemistry 09/2014; 25(12). DOI:10.1016/j.jnutbio.2014.06.014 · 4.59 Impact Factor
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    ABSTRACT: The effect of high hydrostatic pressure (HHP) treatment (150, 300, and 450 MPa for 0, 2.5, and 5 min) on total sodium dodecyl sulfate (SDS)-soluble and sarcoplasmic proteins in frozen (−10 °C for 3 months) Atlantic mackerel (Scomber scombrus) was evaluated. Proteomics tools based on image analysis of SDS-polyacrylamide gel electrophoresis (SDS-PAGE) protein gels and protein identification by tandem mass spectrometry (MS/MS) were applied. Total SDS-soluble proteins, composed in high proportion of myofibrillar proteins, were stable under pressurization treatment in terms of solubility and electrophoretic gel profiles. However, pressurization reduced sarcoplasmic proteins’ solubility, modified their one-dimensional (1-D)/two-dimensional (2-D) SDS-PAGE patterns in a direct-dependent manner, and exerted a selective effect on particular sarcoplasmic proteins depending on processing conditions. Thus, protein bands assigned to creatine kinase, fructose-bisphosphate aldolase A, glycogen phosphorylase, and β-enolase were degraded at 300–450 MPa. Additionally, the stability of triosephosphate isomerase B, phosphoglucomutase, and phosphoglycerate kinase-1 was found to be HHP-reduced when submitted at 450 MPa. HHP processing (300–450 MPa) also induced a cross-linking product formation of pyruvate kinase and two compounds derived from tropomyosin at 450 MPa. Frozen storage time of pressurized samples induced an additional lessening in protein solubility, but electrophoretic patterns were not modified. The present investigation emphasizes the higher lability of sarcoplasmic proteins under HHP treatment and the important role of these proteins in the sensory quality enhancement provided by milder HHP conditions on frozen mackerel. HHP technology is expected to boost the development of novel tailored processing approaches to tackle food quality challenges.
    Food and Bioprocess Technology 08/2014; 7(8). DOI:10.1007/s11947-013-1250-1 · 3.13 Impact Factor
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    44th WEFTA meeting; 06/2014
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    ABSTRACT: The study innovatively pinpoints target proteins of carbonylation, a key PTM induced by oxidative stress, in the SHROB (genetically obese spontaneously hypertensive) rat model of metabolic syndrome (MetS). Protein carbonylation was assessed by a fluorescence-labelling proteomics approach, and complemented with biometric and biochemical markers of MetS. SHROB and healthy Wistar rats were fed two diets, soybean and linseed oil supplementations, in order to distinguish intrinsic carbonylation of SHROB animals from diet-modulated carbonylation unrelated to MetS. First exploratory data showed similar carbonylation patterns and metabolic conditions in SHROB rats fed soybean and linseed, but different from Wistar animals. A total of 18 carbonylated spots in liver, and 12 in skeletal tissue, related to pathways of lipid (29.6%), carbohydrate (25.9%) and amino acid (18.5%) metabolisms, were identified. In particular, SHROB animals present higher carbonylation in four liver proteins belonging to lipid metabolism, redox regulation and chaperone activity (ALDH2, PDI, PDIA3, PECR), and in the skeletal muscle ALDOA involved in muscle dysfunction. Conversely, SHROB rats display lower carbonylation in liver albumin, AKR1C9, ADH1 and catalase. This investigation provides a novel perspective of carbonylation in the context of metabolic disorders, and may be a starting point to characterize new redox pathways exacerbating MetS. Oxidative stress is a concomitant factor in the pathogenesis of MetS that induces oxidative PTM as carbonylation. Through the use a redox proteomics approach, we have thoroughly mapped the occurrence of protein targets of carbonylation in the genetically-induced MetS model SHROB rat. The present research brings a new insight of MetS pathogenesis and it may provide valuable information to understand the biological impact of oxidative stress in patients with MetS.
    Journal of proteomics 04/2014; DOI:10.1016/j.jprot.2014.04.036 · 5.07 Impact Factor
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    ABSTRACT: The present work describes the development of a robust and sensitive targeted analysis platform for the simultaneous quantification in blood plasma of lipid oxygenated mediators and fatty acids using solid-phase extraction (SPE) and high-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS). The concurrent analysis of these lipid mediators is challenging because of their instability, differences in solubility, and the frequent occurrence of isobaric forms with similar fragmentation patterns. Results demonstrated that the reduction of SPE temperature to 4 °C is a critical parameter for preserving the hydroperoxy derivatives. Polymeric HLB cartridges increased 40-50 % ARA, EPA, and DHA sensitivity compared to C18 sorbent and also provided higher global performance for most hydroxides and other oxidation products. The proposed method for the two tested mass analyzers yields high sensitivity, good linearity, and reproducibility, with detection limits ranging 0.002-7 ng/mL and global recoveries as high as 85-112 %. However, the additional advantage of the linear ion trap (LIT) mass analyzer working in full scan product ion mode, compared to the triple quadrupole (QqQ) operating in multiple reaction monitoring (MRM), should be noted: the full scan product ion mode provides the full fragmentation spectra of compounds that allowed the discrimination of coeluting isomers and false positive identifications without additional chromatography development. The proposed lipidomic procedure demonstrates a confident, simple, and sensitive method to profile in plasma a wide range of lipid eicosanoid and docosanoid mediators, including innovatively the analysis of hydroperoxy congeners and nonoxidized PUFA precursors.
    Analytical and Bioanalytical Chemistry 03/2014; 406(12). DOI:10.1007/s00216-014-7701-3 · 3.66 Impact Factor
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    ABSTRACT: This work focuses on the effect of a previous high pressure processing (HPP) on the lipid damage development occurring during the frozen storage (−10°C; up to 3 months) of Atlantic horse mackerel (Trachurus trachurus). HPP conditions included different pressure (150, 300, 450 MPa) and pressure holding time (0.0, 2.5, 5.0 min) values. During frozen storage, horse mackerel muscle was analyzed for lipid hydrolysis (free fatty acid assessment; FFA) and oxidation (formation of peroxides, thiobarbituric acid reactive substances, and fluorescent compounds), and polyene content. An inhibition of lipid hydrolysis development was observed; thus, both an increasing pressure level and pressure holding time led to a marked inhibition of FFA content throughout the frozen storage. Concerning the lipid oxidation development, a partial inhibition was also obtained during the frozen storage (months 1 and 3) by increasing the pressure level applied (namely, fluorescent and peroxide compound formation); however, pressure holding time did not lead to a definite trend. No effect of HPP treatment was concluded on the polyene content of the fish muscle lipids. Present research provides novel information concerning the employment of HPP technology focused on the inhibition of lipid damage during a subsequent frozen storage.Practical applications: Frozen storage of fatty and medium-fat fish species is known to be strongly limited by lipid damage development, which is a drawback to its commercialization as such or to its subsequent employment as raw material in other kinds of processing (canneries, smoking, etc.). Present research provides valuable information concerning the employment of the high pressure technology to inhibit lipid damage development during the subsequent frozen storage of Atlantic horse mackerel (T. trachurus). Thus, both an increasing pressure level (from 150 to 450 MPa) and pressure holding time (from 0 to 5 min) led to a marked inhibition of lipid hydrolysis during frozen storage. Additionally, inhibition of lipid oxidation was produced by increasing the pressure level. Since the response to HPP of marine species has been reported to vary with species, a preliminary study is recommended to be carried out before applying the high pressure-frozen storage combining strategy.
    European Journal of Lipid Science and Technology 12/2013; 115(12). DOI:10.1002/ejlt.201300027 · 2.03 Impact Factor
  • José Manuel Gallardo, Ignacio Ortea, Mónica Carrera
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    ABSTRACT: This review is a critical overview of advances in proteomics applied in food technology, which may be classified into two main topics: (i) authentication of food components as a tool to comply with foodlabeling regulations and policies; and, (ii) food-technology research, mainly for the development of fast, reliable methods to detect and to identify spoilage and/or pathogenic microorganisms in food and for the study of changes in food components as a consequence of food processing. (c) 2013 Elsevier Ltd. All rights reserved.
    TrAC Trends in Analytical Chemistry 12/2013; 52:135–141. DOI:10.1016/j.trac.2013.05.019 · 6.61 Impact Factor
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    ABSTRACT: High consumption of fish carries a lower risk of cardiovascular disease as a consequence of dietary omega-3 long chain polyunsaturated fatty acid (n-3 PUFA; especially EPA and DHA) content. A controversy exists about the component/s responsible of these beneficial effects and, in consequence, which is the best proportion between both fatty acids. We sought to determine, in healthy Wistar rats, the proportions of EPA and DHA that would induce beneficial effects on biomarkers of oxidative stress, and cardiovascular disease risk. Female Wistar rats were fed for 13 weeks with 5 different dietary supplements of oils; 3 derived from fish (EPA/DHA ratios of 1:1, 2:1, 1:2) plus soybean and linseed as controls. The activities of major antioxidant enzymes (SOD, CAT, GPX, and GR) were determined in erythrocytes and liver, and the ORAC test was used to determine the antioxidant capacity in plasma. Also measured were: C reactive protein (CRP), endothelial dysfunction (sVCAM and sICAM), prothrombotic activity (PAI-1), lipid profile (triglycerides, cholesterol, HDLc, LDLc, Apo-A1, and Apo-B100), glycated haemoglobin and lipid peroxidation (LDL-ox and MDA values). After three months of nutritional intervention, we observed statistically significant differences in the ApoB100/ApoA1 ratio, glycated haemoglobin, VCAM-1, SOD and GPx in erythrocytes, ORAC values and LDL-ox. Supplementation with fish oil derived omega-3 PUFA increased VCAM-1, LDL-ox and plasma antioxidant capacity (ORAC). Conversely, the ApoB100/ApoA1 ratio and percentage glycated haemoglobin decreased. Our results showed that a diet of a 1:1 ratio of EPA/DHA improved many of the oxidative stress parameters (SOD and GPx in erythrocytes), plasma antioxidant capacity (ORAC) and cardiovascular risk factors (glycated haemoglobin) relative to the other diets.
    Lipids in Health and Disease 10/2013; 12(1):140. DOI:10.1186/1476-511X-12-140 · 2.31 Impact Factor
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    ABSTRACT: The present study aims to compare two molecular technologies, 16S rRNA sequencing and matrix-assisted laser desorption ionisation time-of-flight mass spectrometry (MALDI TOF - MS), for bacterial species identification in seafood. With this aim, 70 reference strains from culture collections, including important seafood-borne pathogenic and spoilage bacterial species, and 50 strains isolated from commercial seafood products, were analyzed by both techniques. Genomic analysis only identified the species of 50% of the isolated strains, proving to be particularly poor at identifying members of the Pseudomonas and Bacillus genera. In contrast, MALDI-TOF MS fingerprinting identified 76% of the strains at the species level. The mass spectral data were submitted to the SpectraBank database (www.spectrabank.org), making this information available to other researchers. Furthermore, cluster analysis of the peak mass lists was carried out with the web application SPECLUST and the calculated groupings were consistent with results determined by a phylogenetic approach that is based on the 16S rRNA sequences. However, the MALDI-TOF MS analysis demonstrated more discriminating potential that allowed for better classification, especially for the Pseudomonas and Bacillus genera. This is of importance with respect to the varying pathogenic and spoilage character at the intra-genus and intra-species level. In this sense, MALDI-TOF MS demonstrated to be a competent bacterial typing tool that extends phenotypic and genotypic approaches, allowing a more ample classification of bacterial strains.
    Electrophoresis 03/2013; 34(6). DOI:10.1002/elps.201200532 · 3.16 Impact Factor
  • Innovative Food Science & Emerging Technologies 01/2013; · 2.25 Impact Factor
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    ABSTRACT: The present study investigates the susceptibility of individual myofibrillar proteins from mackerel (Scomber scombrus) mince to undergo carbonylation reactions during chilled storage, and the antioxidant capacity of (+)-catechin to prevent oxidative processes of proteins. The carbonylation of each particular protein was quantified by combining the labelling of protein carbonyls by fluorescein-5-thiosemicarbazide (FTSC) with 1-D or 2-D gel electrophoresis. Alpha skeletal actin, glycogen phosphorylase, unnamed protein product (UNP) similar to enolase, pyruvate kinase, isoforms of creatine kinase, aldolase A and an isoform of glyceraldehyde 3-phosphate dehydrogenase (G3PDH) showed elevated oxidation in chilled non-supplemented mince. Myosin heavy chain (MHC) was not carbonylated in chilled muscle, but an extensive MHC degradation was observed in those samples. The supplementation of catechin reduced protein oxidation and lipid oxidation in a concentration-dependent manner: control>25>100≈200ppm. Therefore, the highest catechin concentrations (100 and 200ppm) exhibited the strongest antioxidant activity. Catechin (200ppm) reduced significantly carbonylation of protein spots identified as glycogen phosphorylase, pyruvate kinase muscle isozyme, isoforms of creatine kinase. Conversely, catechin was ineffective to inhibit the oxidation of actin and UNP similar to enolase. These results draw attention to the inefficiency of catechin to prevent actin oxidation, in contrast to the extremely high efficiency of catechin in inhibiting oxidation of lipids and other proteins.
    Food Chemistry 01/2013; 136(1):64-72. DOI:10.1016/j.foodchem.2012.07.109 · 3.26 Impact Factor
  • Mónica Carrera, Benito Cañas, José M Gallardo
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    ABSTRACT: This paper presents the first proteome network map for the sarcoplasmic fish proteome. A total of 183 non-redundant annotated proteins were identified in a shotgun proteome-wide analysis from 15 different fish species. The final protein compilation was investigated by integrated in-silico studies, including functional GO term enrichment, pathways studies and networks analysis. An in-silico interactomics map was built up merging all the identified proteins. The whole confidence network contains 84 nodes and 279 interactions. Most of the sarcoplasmic fish proteins were grouped under pathways and networks referring to energy, catabolism and lipid metabolism. As a new potential nutritional ingredient valuable bioactive peptides were also predicted after an in-silico human gastrointestinal digestion. As is presented in this study, the integrated global proteomics results and the bioinformatics analysis of the sarcoplasmic fish proteome show the feasibility of this approach to provide a comprehensive knowledge of this fraction since a functional and nutritional point of view.
    Journal of proteomics 11/2012; 78. DOI:10.1016/j.jprot.2012.11.016 · 5.07 Impact Factor
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    ABSTRACT: The potential value of different dietary eicosapentaenoic acid (EPA, 20:5) and docosahexaenoic acid (DHA, 22:6) ratios (1:1, 2:1 and 1:2, respectively) on protein redox states from plasma, kidney, skeletal muscle and liver was investigated in Wistar rats. Dietary fish oil groups were compared with animals fed soybean and linseed oils, vegetable oils enriched in omega-6 linoleic acid (LA, 18:2) and omega-3 alpha-linolenic acid (ALA, 18:3), respectively. Fish oil treatments were effective to reduce the level of total fatty acids in plasma, and to enrich the plasmatic free fatty acid fraction and erythrocyte membranes in EPA and DHA. A proteomic approach consisting of fluorescein-5-thiosemicarbazide (FTSC)-labeling of protein carbonyls, FTSC-intensity visualization on 1-DE or 2-DE gels and protein identification by MS/MS was used for the protein oxidation assessment. Albumin was found to be the most carbonylated protein in plasma for all dietary groups, and its oxidation level was significantly modulated by dietary interventions. Supplementations with equal EPA/DHA ratio (1:1) showed the lowest oxidation score for plasma albumin, followed in increasing order of carbonylation by 1:2<2:1≈linseed<soybean. Oxidation patterns of myofibrillar skeletal muscle proteins, and cytosolic proteins from kidney and liver, also indicated a protective effect on proteins for the fish oil treatments, exhibiting the 1:1 ratio the lowest protein oxidation scores. It is remarkable the effect of fish oil treatments in reducing carbonylation on specific proteins from plasma (albumin), skeletal muscle (actin) and liver (albumin, argininosuccinate synthetase, 3-α-hydroxyxteroid dehydrogenase). The present investigation highlights the efficiency of dietary fish oil to reduce in vivo oxidative damage of proteins compared to oils enriched in the 18-carbon polyunsaturated fatty acids (PUFA) omega-3 ALA or omega-6 LA, and such antioxidant activity may differ among different fish oil sources due to variations in EPA/DHA content.
    Free Radical Biology and Medicine 11/2012; DOI:10.1016/j.freeradbiomed.2012.11.004 · 5.27 Impact Factor
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    ABSTRACT: Although seafood species identification has traditionally relied on morphological analysis, sometimes this is difficult to apply for the differentiation among penaeid shrimps owing to their phenotypic similarities and to the frequent removal of external carapace during processing. The objective of this review is to provide an updated and extensive overview on the molecular methods for shrimp and prawn species authentication, in which several omics approaches based on protein and DNA analysis are described. DNA-based methods include the amplification by PCR of different genes, commonly the mitochondrial 16S ribosomal RNA and cytochrome oxidase I genes. A recently described method based on RFLP coupled to PCR turned out to be particularly interesting for species differentiation and origin identification. Protein analysis methods for the characterization and detection of species-specific peptides are also summarized, emphasizing some novel proteomics-based approaches, such as phyloproteomics, peptide fragmentation, and species-specific peptide detection by HPLC coupled to multiple reaction monitoring (MRM) MS, the latter representing the fastest method described to date for species authentication in food.
    Electrophoresis 08/2012; 33(15):2201-11. DOI:10.1002/elps.201100576 · 3.16 Impact Factor
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    ABSTRACT: MALDI-TOF MS has proved to be an accurate, rapid, and cost-effective technique for microbial identification in which the spectral fingerprint of an unknown strain can be compared to a database of spectra from reference strains. Most of the existing databases are private and often costly to access, and little spectral information is shared among researchers. The objective of the present communication is to introduce the SpectraBank database (http://www.spectrabank.org), which provides open access MALDI-TOF mass spectra from a variety of microorganisms. This work aims to familiarize readers with the SpectraBank database, from the sample preparation, data collection, and data analysis to how the spectral reference data can be used for microbial species identification. The database currently includes more than 200 MALDI-TOF MS spectra from more than 70 bacterial species and links to the freely available web-based application SPECLUST (http://bioinfo.thep.lu.se/speclust.html) to allow comparisons of the obtained peak mass lists and evaluate phyloproteomic relationships. The SpectraBank database is intended to be expanded by the addition of new spectra from microbial strains, obtained in our laboratory and by other researchers.
    Electrophoresis 07/2012; 33(14):2138-42. DOI:10.1002/elps.201200074 · 3.16 Impact Factor
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    ABSTRACT: Streptococcus parauberis is known as an etiological agent of mastitis in cows and for producing streptococcosis in farmed fish, although its presence in foods has seldom been reported. In this work, two bacterial isolates were recovered from a spoiled vacuum-packaged refrigerated seafood product. Both isolates were identified by 16S rRNA gene sequencing, exhibiting 99% homology with respect to S. parauberis. Both isolates were also characterized by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS). Genetic analysis revealed the clonal homogeneity of the isolates and their grouping together with other S. parauberis strains in a different cluster with respect to Streptococcus uberis strains. Proteomic analysis by MALDI-TOF MS allowed for the identification of five mass peaks in the range of 2200-6000 m/z that resulted to be specific to the species S. parauberis and allowed its rapid and direct identification with respect to other pathogenic and spoilage bacteria potentially present in seafood and other food products. This study represents, to our knowledge, the first report of S. parauberis in seafood in general and in vacuum-packed food products in particular. Moreover, it provides a rapid method based on MALDI-TOF MS for the identification of S. parauberis.
    Food Microbiology 05/2012; 30(1):91-7. DOI:10.1016/j.fm.2011.10.012 · 3.37 Impact Factor
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    Food Quality, 04/2012; , ISBN: 978-953-51-0560-2

Publication Stats

2k Citations
248.32 Total Impact Points

Institutions

  • 1999–2015
    • Spanish National Research Council
      • • Department of Food Science & Technology
      • • Instituto de Investigaciones Marinas de Vigo
      • • Department of Chemical and Surfactants Technology
      Madrid, Madrid, Spain
  • 2003–2012
    • University of Santiago de Compostela
      • Faculty of Veterinary
      Santiago, Galicia, Spain
  • 2009
    • Marine Environmental Research Institute
      Blue Hills, Connecticut, United States