75 research outputs found

    Report on the Verification of the Performance of MON 87705 and MON 89788 Event-specific PCR-based Methods applied to DNA Extracted from GM Stack Soybean MON 87705 x MON 89788

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    The EU-RL GMFF has previously validated individually, and declared fit for purpose, the detection methods for the single line soybean events MON 87705 and MON 89788 and has published the corresponding reports (see http://gmo-crl.jrc.ec.europa.eu/StatusOfDossiers.aspx). In line with the approach defined by the ENGL (http://gmo-crl.jrc.ec.europa.eu/doc/Min_Perf_Requirements_Analytical_methods.pdf) the EU-RL GMFF therefore has carried out only an in-house verification of the performance of each validated method when applied to DNA extracted from the GM stack MON 87705 x MON 89788 soybean. The hereby reported in-house verification study led to the conclusion that the individual methods meet the ENGL requirements also when applied to DNA extracted from the GM stack MON 87705 x MON 89788 soybean.JRC.I.3-Molecular Biology and Genomic

    Event-specific Method for the Quantification of Soybean Line 40-3-2 Using Real-time PCR - Validation Report and Protocol - Report on the Validation of a DNA Extraction Method for Soybean Seeds

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    The JRC as Community Reference Laboratory for GM Food and Feed (CRL-GMFF), established by Regulation (EC) No 1829/2003, in collaboration with the European Network of GMO Laboratories (ENGL), has carried out a collaborative study to assess the performance of a quantitative event-specific method to detect and quantify the 40-3-2 transformation event in soybean DNA (unique identifier MON-¿4¿32-6). The collaborative trial was conducted according to internationally accepted guidelines (1, 2). In accordance with Regulation (EC) No 1829/2003 of 22 September 2003 on genetically modified food and feed and with Regulation (EC) No 641/2004 of 6 April 2004 on detailed rules for the implementation of Regulation (EC) No 1829/2003, Monsanto provided the detection method and the samples (soybean seeds containing the transformation event and conventional soybean seeds). The JRC prepared the validation samples (calibration samples and blind samples at unknown GM percentage [DNA/DNA]). The collaborative trial involved fourteen laboratories from nine European countries. The results of the international collaborative trial met the ENGL performance requirements and the scientific understanding about satisfactory method performance. Therefore, the CRL-GMFF considers the method validated as fit for the purpose of regulatory compliance. The results of the collaborative study are made publicly available at http://gmo-crl.jrc.it/.JRC.I.6-Biotechnology and GMO

    Event-specific Method for the Quantification of Soybean Line 40-3-2 Using Real-time PCR - Validation Report and Protocol - Report on the Validation of a DNA Extraction Method for Soybean Seeds

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    The JRC as Community Reference Laboratory for GM Food and Feed (CRL-GMFF), established by Regulation (EC) No 1829/2003, in collaboration with the European Network of GMO Laboratories (ENGL), has carried out a collaborative study to assess the performance of a quantitative event-specific method to detect and quantify the 40-3-2 transformation event in soybean DNA (unique identifier MON-¿4¿32-6). The collaborative trial was conducted according to internationally accepted guidelines (1, 2). In accordance with Regulation (EC) No 1829/2003 of 22 September 2003 on genetically modified food and feed and with Regulation (EC) No 641/2004 of 6 April 2004 on detailed rules for the implementation of Regulation (EC) No 1829/2003, Monsanto provided the detection method and the samples (soybean seeds containing the transformation event and conventional soybean seeds). The JRC prepared the validation samples (calibration samples and blind samples at unknown GM percentage [DNA/DNA]). The collaborative trial involved fourteen laboratories from nine European countries. The results of the international collaborative trial met the ENGL performance requirements and the scientific understanding about satisfactory method performance. Therefore, the CRL-GMFF considers the method validated as fit for the purpose of regulatory compliance. The results of the collaborative study are made publicly available at http://gmo-crl.jrc.it/.JRC.I.6-Biotechnology and GMO

    Multiomic studies to improve fruit quality of berry fruits

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    In this study we are going to use different omic-techniques to analyze fruits of three species of berries such as strawberry, raspberry and black currant. Berry fruit are well appreciated for their delicate flavor and nutraceutical properties, with consumer demand increasing over the last years. Furthermore, climate change and market globalization have made necessary to improve the production while maintaining fruit quality traits. Goodberry project is developping analytical platforms, covering from transcriptomic to metabolites and volatile compounds analysis, to find new factors controlling plant adaptation, fruit production and quality. In this study we implement the metabolomic analysis of strawberry, raspberry and black currant fruits from the 2017 harvest, as well as 2018 harvest during this year. To analyze and compare the data we use multiomic tools and bioinformatics to extract properly conclusion The analyses take different berry cultivars, adapted to diverse environments, were grown in 2017 and 2018 in different latitudes (Germany, France, Norway, Italy, Poland and Scotland). The data comes from a combination of gas-chromatography-mass spectrometry (GC-TOF-MS) and headspace solid phase micro extraction (HS-SPME) coupled with GC-MS was used to semi-quantify fruit primary metabolome and volatilome. Around 50 key primary metabolites, including sugars and acids, which are fundamental factors influencing fruit taste and 75 volatiles, responsible of the aroma, were identified across the different genotypes and climates. Multivariate statistical approaches allow us to point out the genetic and environmental factors underlying complex metabolic traits involved in fruit quality. Preliminary analysis showed that both climate and genetic factors influence primary metabolite and volatile content, even if the environment seems to have a stronger impact on the first one.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tec

    Application of multiomic technologies to study the environmental impact on berry fruit quality

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    Berries, such as strawberry, raspberry and black currant, are well appreciated for their delicate flavor and nutraceutical properties, with consumer demand increasing over the last years. However, climate change and market globalization have made necessary to improve the production while maintaining fruit quality traits. Among the EU GoodBerry project’s objetive are develop state-of-the-art analytical platforms, covering from transcriptomic to metabolites and volatile compounds analysis, to find new factors controlling plant adaptation, fruit production and quality and use the data to face climate changes. Here we present the metabolomic analysis of strawberry, raspberry and black currant fruits from the 2017 harvest. Different berry cultivars, adapted to diverse environments, were grown in 2017 and 2018 in different latitudes (Germany, France, Norway, Italy and Poland) combination of spectrometry techniques was used to semi-quantify fruit primary metabolome and volatilome. Around 50 key primary metabolites, including sugars and acids, which are fundamental factors influencing fruit taste and 75 volatiles, responsible of the aroma, were identified across the different genotypes and climates. Multivariate statistical approaches allow us to point out the genetic and environmental factors underlying complex metabolic traits involved in fruit quality.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

    Endoglin, a novel biomarker and therapeutical target to prevent malignant peripheral nerve sheath tumor growth and metastasis.

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    PURPOSE Malignant peripheral nerve sheath tumors (MPNSTs) are highly aggressive soft-tissue sarcomas that lack effective treatments, underscoring the urgent need to uncover novel mediators of MPNST pathogenesis that may serve as potential therapeutic targets. Tumor angiogenesis is considered a critical event in MPNST transformation and progression. Here, we have investigated whether endoglin (ENG), a TGF-β coreceptor with a crucial role in angiogenesis, could be a novel therapeutic target in MPNSTs. EXPERIMENTAL DESIGN ENG expression was evaluated in human peripheral nerve sheath tumor tissues and plasma samples. Effects of tumor cell-specific ENG expression on gene expression, signaling pathway activation and in vivo MPNST growth and metastasis were investigated. The efficacy of ENG targeting in monotherapy or in combination with MEK inhibition was analyzed in xenograft models. RESULTS ENG expression was found to be upregulated in both human MPNST tumor tissues and plasma circulating small extracellular vesicles. We demonstrated that ENG modulates Smad1/5 and MAPK/ERK pathway activation and pro-angiogenic and pro-metastatic gene expression in MPNST cells and plays an active role in tumor growth and metastasis in vivo. Targeting with ENG-neutralizing antibodies (TRC105/M1043) decreased MPNST growth and metastasis in xenograft models by reducing tumor cell proliferation and angiogenesis. Moreover, combination of anti-ENG therapy with MEK inhibition effectively reduced tumor cell growth and angiogenesis. CONCLUSIONS Our data unveil a tumor-promoting function of ENG in MPNSTs and support the use of this protein as a novel biomarker and a promising therapeutic target for this disease.We apologize to those authors whose work could not be cited due to size limitations. We thank Dr. Eduard Serra, Dr. Conxi Lázaro and Dr. David Lyden for their support in the project. We also thank Héctor Tejero for his help in analyzing RNA-seq data. Dr. Peinado laboratory is funded by US Department of Defense (W81XWH-16-1-0131), Agencia Estatal de Investigación/Ministerio de Ciencia e Innovación (AEI/MCIN) (PID2020-118558RB-I00/AEI/10.13039/501100011033), Fundación Proyecto Neurofibromatosis, European Union’s Horizon 2020 research and innovation programme “proEVLifeCycle” under the Marie Skłodowska-Curie grant agreement No 860303, and Fundación Científica AECC. We are also grateful for the support of the Ministerio de Universidades (Programa de Formación de Profesorado Universitario (FPU)) for the fellowship FPU016/05356 awarded to T. González-Muñoz and to the Translational NeTwork for the CLinical application of Extracellular VesicleS (TeNTaCLES) RED2018-102411-T(AEI/10.13039/501100011033). A. Di Giannatale was supported during this work by a research gran Nuovo-Soldati Foundation. The CNIO, certified as Severo Ochoa Excellence Centre, is supported by the Spanish Government through the Instituto de Salud Carlos III.N

    Planck intermediate results XXIV : Constraints on variations in fundamental constants

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    Any variation in the fundamental physical constants, more particularly in the fine structure constant, a, or in the mass of the electron, me, affects the recombination history of the Universe and cause an imprint on the cosmic microwave background angular power spectra. We show that the Planck data allow one to improve the constraint on the time variation of the fine structure constant at redshift z - 10(3) by about a factor of 5 compared to WMAP data, as well as to break the degeneracy with the Hubble constant, H-0. In addition to a, we can set a constraint on the variation in the mass of the electron, me, and in the simultaneous variation of the two constants. We examine in detail the degeneracies between fundamental constants and the cosmological parameters, in order to compare the limits obtained from Planck and WMAP and to determine the constraining power gained by including other cosmological probes. We conclude that independent time variations of the fine structure constant and of the mass of the electron are constrained by Planck to Delta alpha/alpha = (3.6 +/- 3.7) x 10(-3) and Delta m(e)/m(e) = (4 +/- 11) x 10(-3) at the 68% confidence level. We also investigate the possibility of a spatial variation of the fine structure constant. The relative amplitude of a dipolar spatial variation in a (corresponding to a gradient across our Hubble volume) is constrained to be delta alpha/alpha = (-2.4 +/- 3.7) x 10(-2).Peer reviewe

    Planck intermediate results XIV : Dust emission at millimetre wavelengths in the Galactic plane

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    Para-infectious brain injury in COVID-19 persists at follow-up despite attenuated cytokine and autoantibody responses

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    To understand neurological complications of COVID-19 better both acutely and for recovery, we measured markers of brain injury, inflammatory mediators, and autoantibodies in 203 hospitalised participants; 111 with acute sera (1–11 days post-admission) and 92 convalescent sera (56 with COVID-19-associated neurological diagnoses). Here we show that compared to 60 uninfected controls, tTau, GFAP, NfL, and UCH-L1 are increased with COVID-19 infection at acute timepoints and NfL and GFAP are significantly higher in participants with neurological complications. Inflammatory mediators (IL-6, IL-12p40, HGF, M-CSF, CCL2, and IL-1RA) are associated with both altered consciousness and markers of brain injury. Autoantibodies are more common in COVID-19 than controls and some (including against MYL7, UCH-L1, and GRIN3B) are more frequent with altered consciousness. Additionally, convalescent participants with neurological complications show elevated GFAP and NfL, unrelated to attenuated systemic inflammatory mediators and to autoantibody responses. Overall, neurological complications of COVID-19 are associated with evidence of neuroglial injury in both acute and late disease and these correlate with dysregulated innate and adaptive immune responses acutely

    Profilo degli acidi grassi e degli steroli nel plasma di neonati sottoposti a nutrizione parenterale

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    Al fine di fornire un’adeguata nutrizione ai bambini pretermine di peso molto basso (<1250), oltre ad una nutrizione enterale, viene associato anche un supporto nutrizionale per via endovenosa tramite emulsioni lipidiche. Esse permettono di raggiungere un elevato apporto energetico senza un eccessivo carico di glucosio e sono fonte di acidi grassi essenziali. Gli acidi grassi essenziali linoleico (C 18:2 ω6) e α-linolenico (C 18:3 ω3) sono importanti precursori per la sintesi dell’acido arachidonico (ARA C 20:4 ω6), dell’acido eicosapentenoico (EPA C 20:5 ω3) e docosaesaenoico (DHA C 22:6 ω3). La disponibilità di adeguate quantità di acidi grassi è importante per la crescita e lo sviluppo del neonato pretermine: in particolar modo l’ARA e il DHA sono necessari per lo sviluppo neurointellettivo del neonato. Le emulsioni lipidiche contengono anche fitosteroli, steroli vegetali che non vengono sintetizzati nell’uomo ma che vengono assunti tramite la dieta. Alcuni studi indicano i fitosteroli come potenzialmente dannosi per la loro possibile associazione con la colestasi. Lo scopo di questo studio è stato quello di analizzare il profilo lipidico plasmatico (acidi grassi e steroli) di 144 neonati pretermine sottoposti a nutrizione parenterale e randomizzati a ricevere 5 emulsioni lipidiche differenti, di cui 2 contenti olio di pesce (Lipidem® e SMOF®) e 3 convenzionali (Intralipid®, ClinOleic® e Lipofundin®). Il contenuto di acidi grassi e di steroli è stato analizzato tramite analisi cromatografiche (TLC, GC/FID, GC/MS). Dai risultati ottenuti si evince che le emulsioni contenenti olio di pesce determinano una modificazione del profilo lipidico plasmatico. In particolare, esse determinano un aumento significativo degli acidi grassi ω3, specialmente l’EPA e il DHA, e una riduzione significativa dei fitosteroli plasmatici. Ciononostante, non abbiamo riscontrato un’ associazione tra alti livelli di fitosteroli plasmatici e colestasi. Risultano necessari successivi studi di follow-up per valutare lo sviluppo neurointellettivo dei pazienti
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