Background Rosaceae species are economically highly relevant crops. Their cultivation systems are constrained by phytopathogens causing severe losses. Plants respond to invading pathogens through signaling mechanisms, a component of which... more
Background Rosaceae species are economically highly relevant crops. Their cultivation systems are constrained by phytopathogens causing severe losses. Plants respond to invading pathogens through signaling mechanisms, a component of which are of them being plant elicitor peptides (Peps). Exogenous application of Peps activates defense mechanisms and reduces the symptoms of pathogen infection in various pathosystems. We have previously identified the Rosaceae Peps and showed, in an ex vivo system, that their topical application efficiently enhanced resistance to the bacterial pathogen Xanthomonas arboricola pv. pruni (Xap). Results Here we demonstrate the effectiveness of Prunus persica peptides PpPep1 and PpPep2 in protecting peach plants in vivo at nanomolar doses, with 40% reduction of the symptoms following Xap massive infection. We used deep sequencing to characterize the transcriptomic response of peach plants to preventive treatment with PpPep1 and PpPep2. The two peptides ind...
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RESULTS: Aerobic plate counts ranged from < 10 to > 10 9 colony-forming units (CFU) g −1 , with the lowest and highest counts recorded for fruits and sprouts respectively. The highest incidence level of coliforms was found in... more
RESULTS: Aerobic plate counts ranged from < 10 to > 10 9 colony-forming units (CFU) g −1 , with the lowest and highest counts recorded for fruits and sprouts respectively. The highest incidence level of coliforms was found in ready-to-eat vegetables, with up to 65.7% of ...
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Commercialization of genetically modified (GM) plants started about two decades ago. In 2014 10% of the total global crop acreage was sown with GM plants. GM maize is cultured in more than 30 countries, and is one of 4 species with the... more
Commercialization of genetically modified (GM) plants started about two decades ago. In 2014 10% of the total global crop acreage was sown with GM plants. GM maize is cultured in more than 30 countries, and is one of 4 species with the majority of commercial GM plants both in terms of acreage and specific events. To support farmers and consumers freedom of choice, coexistence systems need to be implemented. MON810, expressing the Bacillus thuringiensis Cry1Ab toxin against corn borers, is the only GM maize transformation event cultured in the EU and predominantly in Spain. Numerous conventional and MON810 maize varieties coexist in agricultural fields since 1998; and the proportion of GM fields has continuously increased up to about 70-80% in some zones, particularly in Catalonia, where fields are usually below 1 ha. Conventional agricultural fields in these zones support high GM pressure and thus, they can represent the worst-case scenario in coexistence studies. We monitored adven...
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Plant elicitor peptides (Peps) are endogenous molecules that induce and amplify the first line of inducible plant defense, known as pattern-triggered immunity, contributing to protect plants against attack by bacteria, fungi and... more
Plant elicitor peptides (Peps) are endogenous molecules that induce and amplify the first line of inducible plant defense, known as pattern-triggered immunity, contributing to protect plants against attack by bacteria, fungi and herbivores. Pep topic application and transgenic expression have been found to enhance disease resistance in a small number of model plant-pathogen systems. The action of Peps relies on perception by specific receptors, so displaying a family-specific activity. Recently, the presence and activity of Peps within the Rosaceae has been demonstrated. Here we characterized the population of Pep sequences within the economically important plant family of Rosaceae, with special emphasis on the Amygdaleae and Pyreae tribes, which include the most relevant edible species such as apple, pear and peach, and numerous ornamental and wild species (e.g. photinia, firethorn and hawthorn). The systematic experimental search for Pep and the corresponding precursor PROPEP sequ...
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This review, which has been prepared within the frame of the European Union (EU)-funded project MARLON, surveys the organisation and characteristics of specific livestock and feed production chains (conventional, organic, GM-free) within... more
This review, which has been prepared within the frame of the European Union (EU)-funded project MARLON, surveys the organisation and characteristics of specific livestock and feed production chains (conventional, organic, GM-free) within the EU, with an emphasis on controls, regulations, traceability, and common production practices. Furthermore, an overview of the origin of animal feed used in the EU as well as an examination of the use of genetically modified organisms (GMOs) in feed is provided. From the data, it shows that livestock is traceable at the herd or individual level, depending on the species. Husbandry practices can vary widely according to geography and animal species, whilst controls and checks are in place for notifiable diseases and general health symptoms (such as mortality, disease, productive performance). For feeds, it would be possible only to make coarse estimates, at best, for the amount of GM feed ingredients that an animal is exposed to. Labeling requirem...
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This review explores the possibilities to determine livestock consumption of genetically modified (GM) feeds/ingredients including detection of genetically modified organism (GMO)-related DNA or proteins in animal samples, and the... more
This review explores the possibilities to determine livestock consumption of genetically modified (GM) feeds/ingredients including detection of genetically modified organism (GMO)-related DNA or proteins in animal samples, and the documentary system that is in place for GM feeds under EU legislation. The presence and level of GMO-related DNA and proteins can generally be readily measured in feeds, using established analytical methods such as polymerase chain reaction and immuno-assays, respectively. Various technical challenges remain, such as the simultaneous detection of multiple GMOs and the identification of unauthorized GMOs for which incomplete data on the inserted DNA may exist. Given that transfer of specific GMO-related DNA or protein from consumed feed to the animal had seldom been observed, this cannot serve as an indicator of the individual animal's prior exposure to GM feeds. To explore whether common practices, information exchange and the specific GM feed traceabi...
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It remains controversial whether sentinel lymph node biopsy (SLNB) should be performed before or after neoadjuvant therapy (NAT). We aimed to evaluate the feasibility and accuracy of SLNB before NAT at a single institution, and to... more
It remains controversial whether sentinel lymph node biopsy (SLNB) should be performed before or after neoadjuvant therapy (NAT). We aimed to evaluate the feasibility and accuracy of SLNB before NAT at a single institution, and to determine its relation to patient prognosis. A prospective study of T1c-T2-T3 N0 breast cancer patients, after ultrasound examination, who underwent SLNB prior to NAT. Overall, disease-specific and disease-free survival were calculated by Kaplan-Meier curves. SLNB before NAT was performed in 123 patients from December 2006 to May 2014. The identification rate was 100%. SLNB was positive in 42.3% of cases (27.6% macrometastases). NAT was chemotherapy in 88.6% of cases and endocrine-therapy in 11.4%. Lymphadenectomy was avoided in 72.4% of cases. Median follow-up was 40 months (range 8-100). Overall and disease-free survival was 90.2% and 88.6% respectively.SLN involvement was not related to patient outcome (p 0.72); however there were significant difference...
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Fruit crops are regarded as important health promoters and constitute a major part of global agricultural production, and Rosaceae species are of high economic impact. Their culture is threatened by bacterial diseases, whose control is... more
Fruit crops are regarded as important health promoters and constitute a major part of global agricultural production, and Rosaceae species are of high economic impact. Their culture is threatened by bacterial diseases, whose control is based on preventative treatments using compounds of limited efficacy and negative environmental impact. One of the most economically relevant examples is the pathogen Xanthomonas arboricola pv. pruni (Xap) affecting Prunus spp. The plant immune response against pathogens can be triggered and amplified by plant elicitor peptides (Peps), perceived by specific receptors (PEPRs). Although they have been described in various angiosperms, scarce information is available on Rosaceae species. Here, we identified the Pep precursor (PROPEP), Pep and PEPR orthologues of 10 Rosaceae species and confirmed the presence of the Pep/PEPR system in this family. We showed the perception and elicitor activity of Rosaceae Peps using the Prunus-Xap pathosystem as proof-of-concept. Treatment with nanomolar doses of Peps induced the corresponding PROPEP and a set of defence-related genes in Prunus leaves, and enhanced resistance against Xap. Peps from the same species had the highest efficiencies. Rosaceae Peps could potentially be used to develop natural, targeted and environmentally friendly strategies to enhance the resistance of Prunus species against biotic attackers.
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The GRACE (GMO Risk Assessment and Communication of Evidence; www.grace-fp7.eu ) project was funded by the European Commission within the 7th Framework Programme. A key objective of GRACE was to conduct 90-day animal feeding trials,... more
The GRACE (GMO Risk Assessment and Communication of Evidence; www.grace-fp7.eu ) project was funded by the European Commission within the 7th Framework Programme. A key objective of GRACE was to conduct 90-day animal feeding trials, animal studies with an extended time frame as well as analytical, in vitro and in silico studies on genetically modified (GM) maize in order to comparatively evaluate their use in GM plant risk assessment. In the present study, the results of a 1-year feeding trial with a GM maize MON810 variety, its near-isogenic non-GM comparator and an additional conventional maize variety are presented. The feeding trials were performed by taking into account the guidance for such studies published by the EFSA Scientific Committee in 2011 and the OECD Test Guideline 452. The results obtained show that the MON810 maize at a level of up to 33 % in the diet did not induce adverse effects in male and female Wistar Han RCC rats after a chronic exposure.
Research Interests: Risk assessment, Animal Feed, Female, Animals, Male, and 3 moreHealth Status, Zea mays, and Risk Assessment
Polymerase chain reaction (PCR) methods are very useful techniques for the detection and quantification of genetically modified organisms (GMOs) in food samples. These methods rely on the amplification of transgenic sequences and... more
Polymerase chain reaction (PCR) methods are very useful techniques for the detection and quantification of genetically modified organisms (GMOs) in food samples. These methods rely on the amplification of transgenic sequences and quantification of the transgenic DNA by comparison to an amplified reference gene. Reported here is the development of specific primers for the rapeseed (Brassica napus) BnACCg8 gene and PCR cycling conditions suitable for the use of this sequence as an endogenous reference gene in both qualitative and quantitative PCR assays. Both methods were assayed with 20 different rapeseed varieties, and identical amplification products were obtained with all of them. No amplification products were observed when DNA samples from other Brassica species, Arabidopsis thaliana, maize, and soybean were used as templates, which demonstrates that this system is specific for rapeseed. In real-time quantitative PCR analysis, the detection limit was as low as 1.25 pg of DNA, which indicates that this method is suitable for use in processed food samples which contain very low copies of target DNA.
Research Interests: Engineering, Reference, Quality Control, Food Analysis, Quantization, and 13 moreMethod, Polymerase Chain Reaction, Gene, Agricultural, CHEMICAL SCIENCES, Gen, Brassica rapa, Agricultural and Food Chemistry, Species Specificity, Sensitivity and Specificity, Genetically Modified Organism (GMO), Acetyl CoA Carboxylase, and Reference standards
Research Interests: Technology, Western Europe, European Union, Quantitative PCR, Biological Sciences, and 16 morereal time PCR, Humans, Maize, Calibration, Animals, Polymerase Chain Reaction, GMO, Zea mays, Genomic DNA, Genome sequence, Transgenic, Certified Reference Material, Base Sequence, Transgenic plant, Molecular Sequence Data, and Reference standards
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Worldwide maize is the second major agricultural commodity and around one-fourth is currently biotech, with significant application of the insect resistant event MON810 particularly in the European Union. Grains are the major... more
Worldwide maize is the second major agricultural commodity and around one-fourth is currently biotech, with significant application of the insect resistant event MON810 particularly in the European Union. Grains are the major commercialized part of the plant, and can be harvested after maturity (for food and feed purposes) or at late milky-starchy stage (for forage uses, with the whole plant). We assessed possible proteomic unintended effects of the MON810 transgene using two-dimensional gel electrophoresis coupled to mass spectrometry. To keep in a realistic scenario we used plants grown in agricultural fields in a region where ~50% of maize was MON810, and analyzed grains at milky-starchy stage. In maize, differential transcripts and metabolites between GM and comparable non-GM varieties tend to be variety specific. Thus, we analyzed two variety pairs, DKC6575/Tietar and PR33P67/PR33P66 which are considered representative of Food and Agriculture Organization 700 and 600 varieties commercially grown in the region. MON810 and non-GM milky-starchy grains had virtually identical proteomic patterns, with a very small number of spots showing fold-variations in the 1-1.8 range. They were all variety specific and had divergent identities and functions. Although 2DE allows the analysis of a limited dataset our results support substantial equivalence between MON810 and comparable non-GM varieties.
Research Interests: Technology, Mass Spectrometry, Proteomics, Agriculture, European Union, and 13 moreBiological Sciences, Maize, Insect Resistance, Zea mays, Genetic variation, Proteome analysis, Transgenic, Food and Agriculture Organization, Gel electrophoresis, Two-Dimensional Gel Electrophoresis, Two dimensional Gel Electrophoresis, Proteome, and Genetically Modified Organism (GMO)
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In this paper, a method for the specific detection and quantification of potato and tomato DNA in food samples with the use of conventional and real-time polymerase chain reaction (PCR) is described. This method is adequate for use in... more
In this paper, a method for the specific detection and quantification of potato and tomato DNA in food samples with the use of conventional and real-time polymerase chain reaction (PCR) is described. This method is adequate for use in food quality routine assays involving highly processed samples for which very tiny amounts of DNA are expected. Detection was achieved by amplifying a region of the metallo-carboxypeptidase inhibitor gene from either the potato (PCI) or the tomato (MCPI) and by using specific primers complementary to the propeptide regions of these inhibitors, which were found to differ for the potato and tomato proproteins. Conventional and real-time PCR systems were based on the same potato- or tomato-specific primer pairs, and quantification was carried out with a TaqMan chemistry-based probe. The methods developed proved to be very specific and sensitive and highly reliable for the identification and quantification of DNA from both plant species. In addition, the construction of plasmids pPAT and pTOM, suitable for use as external calibration standards for the elaboration of comparative amplification profiles, is reported. Limits of detection and quantification with the use of these plasmid standards are given. Specificity and copy number conservation among different cultivars were analyzed, and the reliability of these systems was tested through their application to the analysis of commercial food samples including potato and/or tomato as components.
Research Interests: Molecular Biology, Multidisciplinary, Quality Control, Quantitative analysis, Calibration, and 12 morePolymerase Chain Reaction, Protease Inhibitors, Food Contamination, Quantitative Analysis, Solanum Tuberosum, Base Sequence, Sensitivity and Specificity, PLANT PROTEINS, Potato Starch, Lycopersicon esculentum, Reference standards, and food handling
A real-time polymerase chain reaction (QPCR) assay was developed for quantitative detection of a genetically modified (GM) maize event TC-1507 and modification to conventional PCR for qualitative purposes. Sequences... more
A real-time polymerase chain reaction (QPCR) assay was developed for quantitative detection of a genetically modified (GM) maize event TC-1507 and modification to conventional PCR for qualitative purposes. Sequences 5&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39;-flanking TC-1507 full-length insert were characterized and showed multiple rearrangements involving insert and maize chloroplast fragments. The event specificity of the TC-1507 assays was based on the detection of transgene and plant rearranged sequences found to 5&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;#39; flank the insertion site. They were fully specific and exhibited a limit of detection below 10 target copies, allowing consistent detection of 0.1% GM levels. The QPCR was highly linear and efficient and proved adequate for quantification of GM contents, aiming at the fulfillment of legal requirements established in the European Union (i.e., compulsory labeling of TC-1507 levels &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;gt;0.9%). It satisfactorily determined TC-1507 contents on different matrixes and was successfully transferred a different laboratory.
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Research Interests: Genetics, Principal Component Analysis, Plant Biology, Agriculture, Plant Molecular Biology, and 17 moreGene expression, Transcriptome, Maize, Nitrogen, Transgenic Crops, Nitrogen Fertilizer, Zea mays, Environmental Conditions, Fertilizers, Natural Variation, Species Specificity, Genetically Modified Organism (GMO), Endotoxins, Biochemistry and cell biology, Safety Assessment, Gene expression profiling, and Differential expression
Molecular evidence has linked the pathophysiology of lymphangioleiomyomatosis (LAM) to that of metastatic breast cancer. Following on this observation, we assessed the association between LAM and subsequent breast cancer. An... more
Molecular evidence has linked the pathophysiology of lymphangioleiomyomatosis (LAM) to that of metastatic breast cancer. Following on this observation, we assessed the association between LAM and subsequent breast cancer. An epidemiological study was carried out using three LAM country cohorts, from Japan, Spain, and the United Kingdom. The number of incident breast cancer cases observed in these cohorts was compared with the number expected on the basis of the country-specific incidence rates for the period 2000-2014. Immunohistochemical studies and exome sequence analysis were performed in two and one tumors, respectively. All cohorts revealed breast cancer standardized incidence ratios (SIRs) ≥ 2.25. The combined analysis of all cases or restricted to pre-menopausal age groups revealed significantly higher incidence of breast cancer: SIR = 2.81, 95 % confidence interval (CI) = 1.32-5.57, P = 0.009; and SIR = 4.88, 95 % CI = 2.29-9.99, P = 0.0007, respectively. Immunohistochemical...
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Para-aortic lymphadenectomy (PAL) is a challenging procedure, performed by minimally invasive surgery in very few centers due to its intrinsic technical complexity. We aim to describe and asses the feasibility and learning curve of... more
Para-aortic lymphadenectomy (PAL) is a challenging procedure, performed by minimally invasive surgery in very few centers due to its intrinsic technical complexity. We aim to describe and asses the feasibility and learning curve of robotic double docking transperitoneal infrarenal PAL combined with oncological pelvic surgery. From March 2010 to May 2013, 50 patients who underwent this procedure using the Da Vinci S surgical system were included. The mean PAL surgery time was 76 min (range 32-150), and mean number of lymph nodes 11.8 (range 1-44). There were no conversions to laparotomy or laparoscopy. The mean hospital stay was 2 days (range 1-25). Statistically significant decreases were noted for mean table rotation time (17 ± 6.8 SD vs. 13 ± 3.6 SD min; p= 0.02) and mean PAL operating time (85,4 ± 25,8 SD vs. 69,8 ± 24,6 SD min; p= 0,04) when comparing the first 20 patients to the rest of patients. . The number of nodes remained similar for the first 20 to the rest of the patients. Double docking transperitoneal infrarenal PAL technique combined with oncological pelvic surgery is feasible with minimal morbidity and a short learning curve.
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Cationic α-helical antimicrobial peptides such as BP100 are of increasing interest for developing novel phytosanitary or therapeutic agents and products with industrial applications. Biotechnological production of these peptides in plants... more
Cationic α-helical antimicrobial peptides such as BP100 are of increasing interest for developing novel phytosanitary or therapeutic agents and products with industrial applications. Biotechnological production of these peptides in plants can be severely impaired due to the toxicity exerted on the host by high-level expression. This can be overcome by using inducible promoters with extremely low activity throughout plant development, although the yields are limited. We examined the use of modified atmospheres using the increased levels of [CO2], commonly used in the food industry, as the inductor agent to biotechnologically produce phytotoxic compounds with higher yields. Here we show that 30 % [CO2] triggered a profound transcriptional response in rice leaves, including a change in the energy provision from photosynthesis to glycolysis, and the activation of stress defense mechanisms. Five genes with central roles in up-regulated pathways were initially selected and their promoters successfully used to drive the expression of phytotoxic BP100 in genetically modified (GM) rice. GM plants had a normal phenotype on development and seed production in non-induction conditions. Treatment with 30 % [CO2] led to recombinant peptide accumulation of up to 1 % total soluble protein when the Os.hb2 promoter was used. This is within the range of biotechnological production of other peptides in plants. Using BP100 as a proof-of-concept we demonstrate that very high [CO2] can be considered an economically viable strategy to drive production of recombinant phytotoxic antimicrobial peptides in plant biofactories.
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Temporary employment accounts in Spain for about one third of total salaried employment since the mid-Eighties, which triples European figures. In the Nineties, two labour market reforms were implemented in an attempt to reduce the very... more
Temporary employment accounts in Spain for about one third of total salaried employment since the mid-Eighties, which triples European figures. In the Nineties, two labour market reforms were implemented in an attempt to reduce the very high incidence of temporary employment, achieving some positive effects on employment. However, we still have a very limited knowledge of the likely effects on
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Measuring microbiological contamination in fruit and vegetables M. Pla, D. Rodriguez-Lazaro, E. Badosa and E. Montesinos, Institute of Food and Agricultural Technology (INTEA), University of Girona, Spain 3.1 Introduction Foodborne... more
Measuring microbiological contamination in fruit and vegetables M. Pla, D. Rodriguez-Lazaro, E. Badosa and E. Montesinos, Institute of Food and Agricultural Technology (INTEA), University of Girona, Spain 3.1 Introduction Foodborne diseases are among the most serious ...
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Genetically modified (GM) crops have been commercially grown for two decades. GM maize is one of 3 species with the highest acreage and specific events. Many countries established a mandatory labeling of products containing GM material,... more
Genetically modified (GM) crops have been commercially grown for two decades. GM maize is one of 3 species with the highest acreage and specific events. Many countries established a mandatory labeling of products containing GM material, with thresholds for adventitious presence, to support consumers' freedom of choice. In consequence, coexistence systems need to be introduced to facilitate commercial culture of GM and non-GM crops in the same agricultural area. On modeling adventitious GM cross-pollination distribution within maize fields, we deduced a simple equation to estimate overall GM contents (%GM) of conventional fields, irrespective of its shape and size, and with no previous information on possible GM pollen donor fields. A sampling strategy was designed and experimentally validated in 19 agricultural fields. With 9 samples, %GM quantification requires just one analytical GM determination while identification of the pollen source needs 9 additional analyses. A decision...