LC-MS/MS method for the determination of organophosphorus pesticides and their metabolites in salmon and zebrafish fed with plant-based feed ingredients

This is a pre-print of an article published in Analytical and Bioanalytical Chemistry. The final authenticated version is available online at:https://doi.org/10.1007/s00216-019-02104-6The composition of Atlantic salmon feed has changed considerably over the last two decades from being marine-based (fishmeal and fish oil) to mainly containing plant ingredients. Consequently, concern related to traditional persistent contaminants typically associated with fish-based feed has been replaced by other potential contaminants not previously associated with salmon farming. This is the case for many pesticides, which are used worldwide to increase food production, and may be present in plant ingredients. Earlier studies have identified two organophosphorus pesticides, chlorpyrifos-methyl and pirimiphos-methyl, in plant ingredients used for aquafeed production. In the present study, we developed a reliable and sensitive analytical method, based on liquid chromatography coupled to tandem mass spectrometry, for the determination of these pesticides and their main metabolites in warm water (zebrafish) and cold water (Atlantic salmon) species, where possible differences in metabolites could be expected. The method was tested in whole zebrafish and in different salmon tissues, such as muscle, bile, kidney, fat, and liver. The final objective of this work was to assess kinetics of chlorpyrifos-methyl and pirimiphos-methyl and their main metabolites in fish tissue, in order to fill the knowledge gaps on these metabolites in fish tissues when fed over prolonged time

Results of the engineering run of the coherent neutrino nucleus interaction experiment (CONNIE)

The CONNIE detector prototype is operating at a distance of 30 m from the core of a 3.8 GWth nuclear reactor with the goal of establishing Charge-Coupled Devices (CCD) as a new technology for the detection of coherent elastic neutrino-nucleus scattering. We report on the results of the engineering run with an active mass of 4 g of silicon. The CCD array is described, and the performance observed during the first year is discussed. A compact passive shield was deployed around the detector, producing an order of magnitude reduction in the background rate. The remaining background observed during the run was stable, and dominated by internal contamination in the detector packaging materials. The in-situ calibration of the detector using X-ray lines from fluorescence demonstrates good stability of the readout system. The event rates with the reactor ON and OFF are compared, and no excess is observed coming from nuclear fission at the power plant. The upper limit for the neutrino event rate is set two orders of magnitude above the expectations for the standard model. The results demonstrate the cryogenic CCD-based detector can be remotely operated at the reactor site with stable noise below2 e RMS and stable background rates. The success of the engineering test provides a clear path for the upgraded 100 g detector to be deployed during 2016.Fil: Aguilar Arevalo, A.. Universidad Nacional Autónoma de México; MéxicoFil: Bertou, Xavier Pierre Louis. Comisión Nacional de Energía Atómica; Argentina. Comisión Nacional de Energía Atómica. Fundación José A. Balseiro; ArgentinaFil: Bonifazi, C.. Universidade Federal do Rio de Janeiro; BrasilFil: Butner, M.. Fermi National Accelerator Laboratory; Estados UnidosFil: Cancelo, G.. Fermi National Accelerator Laboratory; Estados UnidosFil: Castañeda Vazquez, A.. Universidad Nacional Autónoma de México; MéxicoFil: Cervantes Vergara, B.. Universidad Nacional Autónoma de México; MéxicoFil: Chavez, C. R.. Universidad Nacional de Asunción; ParaguayFil: Da Motta, H.. Centro Brasileiro de Pesquisas Físicas; BrasilFil: D'Olivo, J. C.. Universidad Nacional Autónoma de México; MéxicoFil: Dos Anjos, J.. Centro Brasileiro de Pesquisas Físicas; BrasilFil: Estrada, J.. Fermi National Accelerator Laboratory; Estados UnidosFil: Fernández Moroni, Guillermo. Universidad Nacional del Sur. Departamento de Ingeniería Eléctrica y de Computadoras. Instituto ; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Ford, R.. Fermi National Accelerator Laboratory; Estados UnidosFil: Foguel, A.. Centro Brasileiro de Pesquisas Físicas; Brasil. Universidade Federal do Rio de Janeiro; BrasilFil: Hernandez Torres, K. P.. Universidad Nacional Autónoma de México; MéxicoFil: Izraelevitch, F.. Fermi National Accelerator Laboratory; Estados UnidosFil: Kavner, A.. University of Michigan; Estados UnidosFil: Kilminster, B.. Universitat Zurich; SuizaFil: Kuk, K.. Fermi National Accelerator Laboratory; Estados UnidosFil: Lima Jr, H. P.. Centro Brasileiro de Pesquisas Físicas; BrasilFil: Makler, M.. Centro Brasileiro de Pesquisas Físicas; BrasilFil: Molina, J.. Universidad Nacional de Asunción; ParaguayFil: Moreno Granados, G.. Universidad Nacional Autónoma de México; MéxicoFil: Moro, Juan Manuel. Universidad Nacional del Sur. Departamento de Ingeniería; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Paolini, Eduardo Emilio. Universidad Nacional del Sur. Departamento de Ingeniería Eléctrica y de Computadoras. Instituto ; ArgentinaFil: Sofo Haro, Miguel Francisco. Comision Nacional de Energia Atomica. Gerencia D/area de Energia Nuclear; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Tiffenberg, Javier Sebastian. Fermi National Accelerator Laboratory; Estados Unidos. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Trillaud, F.. Universidad Nacional Autónoma de México; MéxicoFil: Wagner, S.. Centro Brasileiro de Pesquisas Físicas; Brasil. Pontificia Universidade Católica do Rio Grande do Sul; Brasi

Comprehensive strategy for pesticide residue analysis through the production cycle of gilthead sea bream and Atlantic salmon

Plant ingredients and processed animal proteins are alternative feedstuffs for fish feeds in aquaculture. However, their use can introduce contaminants like pesticides that are not previously associated with marine Atlantic salmon and gilthead sea bream farming. This study covers the screening of around 800 pesticides by gas chromatography (GC) and liquid chromatography (LC) coupled to high resolution time-of-flight mass spectrometry in matrices throughout the entire marine food production chain. Prior to analysis of real-world samples, the screening methodology was validated for 252 pesticides to establish the screening detection limit. This was 0.01 mg kg−1 for 113 pesticides (45%), 0.05 mg kg−1 for 73 pesticides (29%) and >0.05 mg kg−1 for 66 pesticides (26%). After that, a quantitative methodology based on GC coupled to tandem mass spectrometry with atmospheric pressure chemical ionization source (GC-APCI-MS/MS) was optimized for the pesticides found in the screening. Although several polar pesticides, of which pirimiphos methyl and chlorpyriphos-methyl were most dominant, were found in plant material and feeds based on these ingredients, none of them were observed in fillets of Atlantic salmon and gilthead sea bream fed on these feeds.This work has been developed within the framework of the Research Unit of Marine Ecotoxicology (IATS (CSIC)-IUPA (UJI)

Colombian essential oil of ruta graveolens against nosocomial antifungal resistant candida strains

Drug resistance in antifungal therapy, a problem unknown until a few years ago, is increasingly assuming importance especially in immunosuppressed patients and patients receiving chemotherapy and radiotherapy. In the past years, the use of essential oils as an approach to improve the effectiveness of antifungal agents and to reduce antifungal resistance levels has been proposed. Our research aimed to evaluate the antifungal activity of Colombian rue, Ruta graveolens, essential oil (REO) against clinical strains of Candida albicans, Candida parapsilopsis, Candida glabrata, and Candida tropicalis. Data obtained showed that C. tropicalis and C. albicans were the most sensitive strains showing minimum inhibitory concentrations (MIC) of 4.1 and 8.2 µg/mL of REO. Time–kill kinetics assay demonstrated that REO showed a fungicidal effect against C. tropicalis and a fungistatic effect against C. albicans. In addition, an amount of 40% of the biofilm formed by C. albicans was eradicated using 8.2 µg/mL of REO after 1 h of exposure. The synergistic effect of REO together with some antifungal compounds was also investigated. Fractional inhibitory concentration index (FICI) showed synergic effects of REO combined with amphotericin B. REO Lead a disruption in the cellular membrane integrity, consequently resulting in increased intracellular leakage of the macromolecules, thus confirming that the plasma membrane is a target of the mode of action of REO against C. albicans and C. tropicalis

Intentional Storytelling to Sustain Low-cost/Free Breast Cancer Services: A Latina Example of Community-driven Advocacy

Background: Community-based public health advocacy efforts are crucial to sustaining the low-cost/free breast cancer services that support underserved populations. Objectives: We introduce two ways in which narrative theory may be a useful tool for developing advocacy materials and provide an example, using a community–academic partnership to promote Latina breast health in Chicago, Illinois. Methods: Community and academic partners 1) engaged 25 Spanish-speaking Latinas in an advocacy workshop, 2) leveraged narrative theory to develop multi-media advocacy materials, and 3) disseminated materials to policymakers. Lessons Learned: Our project highlights 1) that narrative theory may be useful to describe how Latinas engage policy-makers in relation to their needs and cultural norms, 2) the importance of flexibility and offering community members multiple options to engage policymakers, and 3) the importance of leveraging partners’ complementary strengths. Conclusions: Narrative theory may be a useful tool for developing advocacy materials in community–academic partnerships

Differential Role of Human Choline Kinase α and β Enzymes in Lipid Metabolism: Implications in Cancer Onset and Treatment

11 pages, 6 figures, 1 table.Background The Kennedy pathway generates phosphocoline and phosphoethanolamine through its two branches. Choline Kinase (ChoK) is the first enzyme of the Kennedy branch of synthesis of 1phosphocholine, the major component of the plasma membrane. ChoK family of proteins is composed by ChoKα and ChoKβ isoforms, the first one with two different variants of splicing. Recently ChoKα has been implicated in the carcinogenic process, since it is over-expressed in a variety of human cancers. However, no evidence for a role of ChoKβ in carcinogenesis has been reported. Methodology/Principal Findings Here we compare the in vitro and in vivo properties of ChoKα1 and ChoKβ in lipid metabolism, and their potential role in carcinogenesis. Both ChoKα1 and ChoKβ showed choline and ethanolamine kinase activities when assayed in cell extracts, though with different affinity for their substrates. However, they behave differentially when overexpressed in whole cells. Whereas ChoKβ display an ethanolamine kinase role, ChoKα1 present a dual choline/ethanolamine kinase role, suggesting the involvement of each ChoK isoform in distinct biochemical pathways under in vivo conditions. In addition, while overexpression of ChoKα1 is oncogenic when overexpressed in HEK293T or MDCK cells, ChoKβ overexpression is not sufficient to induce in vitro cell transformation nor in vivo tumor growth. Furthermore, a significant upregulation of ChoKα1 mRNA levels in a panel of breast and lung cancer cell lines was found, but no changes in ChoKβ mRNA levels were observed. Finally, MN58b, a previously described potent inhibitor of ChoK with in vivo antitumoral activity, shows more than 20-fold higher efficiency towards ChoKα1 than ChoKβ. Conclusion/Significance This study represents the first evidence of the distinct metabolic role of ChoKα and ChoKβ isoforms, suggesting different physiological roles and implications in human carcinogenesis. These findings constitute a step forward in the design of an antitumoral strategy based on ChoK inhibition.This work has been supported by grants to JCL from Comunidad de Madrid (GR-SAL-0821-2004), Ministerio de Ciencia e Innovación (SAF2008-03750, RD06/0020/0016), Fundación Mutua Madrileña, and by a grant to ARM from Fundación Mutua Madrileña.Peer reviewe

GSE4‐loaded nanoparticles a potential therapy for lung fibrosis that enhances pneumocyte growth, reduces apoptosis and DNA damage

Idiopathic pulmonary fibrosis is a lethal lung fibrotic disease, associated with aging with a mean survival of 2-5 years and no curative treatment. The GSE4 peptide is able to rescue cells from senescence, DNA and oxidative damage, inflammation, and induces telomerase activity. Here, we investigated the protective effect of GSE4 expression in vitro in rat alveolar epithelial cells (AECs), and in vivo in a bleomycin model of lung fibrosis. Bleomycin-injured rat AECs, expressing GSE4 or treated with GSE4-PLGA/PEI nanoparticles showed an increase of telomerase activity, decreased DNA damage, and decreased expression of IL6 and cleaved-caspase 3. In addition, these cells showed an inhibition in expression of fibrotic markers induced by TGF-β such as collagen-I and III among others. Furthermore, treatment with GSE4-PLGA/PEI nanoparticles in a rat model of bleomycin-induced fibrosis, increased telomerase activity and decreased DNA damage in proSP-C cells. Both in preventive and therapeutic protocols GSE4-PLGA/PEI nanoparticles prevented and attenuated lung damage monitored by SPECT-CT and inhibited collagen deposition. Lungs of rats treated with bleomycin and GSE4-PLGA/PEI nanoparticles showed reduced expression of α-SMA and pro-inflammatory cytokines, increased number of pro-SPC-multicellular structures and increased DNA synthesis in proSP-C cells, indicating therapeutic efficacy of GSE4-nanoparticles in experimental lung fibrosis and a possible curative treatment for lung fibrotic patients

Secretion of biologically active interferon-gamma inducible protein-10 (IP-10) by Lactococcus lactis

<p>Abstract</p> <p>Background</p> <p>Chemokines are a large group of chemotactic cytokines that regulate and direct migration of leukocytes, activate inflammatory responses, and are involved in many other functions including regulation of tumor development. Interferon-gamma inducible-protein-10 (IP-10) is a member of the C-X-C subfamily of the chemokine family of cytokines. IP-10 specifically chemoattracts activated T lymphocytes, monocytes, and NK cells. IP-10 has been described also as a modulator of other antitumor cytokines. These properties make IP-10 a novel therapeutic molecule for the treatment of chronic and infectious diseases. Currently there are no suitable live biological systems to produce and secrete IP-10. <it>Lactococcus lactis </it>has been well-characterized over the years as a safe microorganism to produce heterologous proteins and to be used as a safe, live vaccine to deliver antigens and cytokines of interest. Here we report a recombinant strain of <it>L. lactis </it>genetically modified to produce and secrete biologically active IP-10.</p> <p>Results</p> <p>The IP-10 coding region was isolated from human cDNA and cloned into an <it>L. lactis </it>expression plasmid under the regulation of the pNis promoter. By fusion to the usp45 secretion signal, IP-10 was addressed out of the cell. Western blot analysis demonstrated that recombinant strains of <it>L. lactis </it>secrete IP-10 into the culture medium. Neither degradation nor incomplete forms of IP-10 were detected in the cell or supernatant fractions of <it>L. lactis</it>. In addition, we demonstrated that the NICE (nisin-controlled gene expression) system was able to express IP-10 "de novo" even two hours after nisin removal. This human IP-10 protein secreted by <it>L. lactis </it>was biological active as demonstrated by Chemotaxis assay over human CD3+T lymphocytes.</p> <p>Conclusion</p> <p>Expression and secretion of mature IP-10 was efficiently achieved by <it>L. lactis </it>forming an effective system to produce IP-10. This recombinant IP-10 is biologically active as demonstrated by its ability to chemoattract human CD3+ T lymphocytes. This strain of recombinant <it>L. lactis </it>represents a potentially useful tool to be used as a live vaccine <it>in vivo</it>.</p

The ALMA-ALPINE [CII] survey: sub-kpc morphology of 3 main-sequence galaxy systems at z~4.5 revealed by ALMA

Context: From redshift 6 to redshift $\approx$ 4 galaxies grow rapidly from low mass galaxies towards the more mature massive galaxies we see at the cosmic noon. Growth via gas accretion and mergers undoubtedly shape this evolution - however, there currently exists much uncertainty over the contribution of each of these processes to the overall evolution of galaxies. Furthermore, previous characterisations of the morphology of galaxies in the molecular gas phase has been limited by the coarse resolution of previous observations. Aims: The goal of this paper is to derive the morpho-kinematic properties of 3 main-sequence systems at $z\sim4.5$, drawn from the ALPINE survey, using brand new high-resolution ALMA data in band 7. The objects were previously characterised as one merger with three components, and and two dispersion-dominated galaxies. Methods: We use intensity and velocity maps, position-velocity diagrams and radial profiles of [CII], in combination with dust continuum maps, to analyse the morphology and kinematics of the 3 systems.} Results: In general, we find that the high-resolution ALMA data reveal more complex morpho-kinematic properties. We identify in one galaxy interaction-induced clumps, showing the profound effect that mergers have on the molecular gas in galaxies, consistent with what is suggested in recent simulations. A galaxy that was previously classified as dispersion dominated turned out to show two bright [CII] emission regions, that could either be merging galaxies or massive star-forming regions within the galaxy itself. The high resolution data for the other dispersion dominated object also revealed clumps of [CII] that were not previously identified. Within the sample, we might also detect star-formation powered outflows (or outflows from Active Galactic Nuclei) which appear to be fuelling diffuse gas regions and enriching the circumgalactic medium.Comment: Submitted to A&A, 13 page