Suchergebnisse

Colorectal cancer is one of the main causes of cancer death worldwide, and novel biomarkers are urgently needed for its early diagnosis and treatment. The utilization of metabolomics to identify and quantify metabolites in body fluids may allow the detection of changes in their concentrations that could serve as diagnostic markers for colorectal cancer and may also represent new therapeutic targets. Metabolomics generates a pathophysiological 'fingerprint' that is unique to each individual. The purpose of our study was to identify a differential metabolomic signature for metastatic colorectal cancer. Serum samples from 60 healthy controls and 65 patients with metastatic colorectal cancer were studied by liquid chromatography coupled to high-resolution mass spectrometry in an untargeted metabolomic approach. Multivariate analysis revealed a separation between patients with metastatic colorectal cancer and healthy controls, who significantly differed in serum concentrations of one endocannabinoid, two glycerophospholipids, and two sphingolipids. These findings demonstrate that metabolomics using liquid-chromatography coupled to high-resolution mass spectrometry offers a potent diagnostic tool for metastatic colorectal cancer. ; This study was supported by a grant (n° 15CC056/DTS17/00081- ISCIII-FEDER) from the Fundación para la Investigación Biosanitaria de Andalucía Oriental (FIBAO) and Roche Pharma S.L. Authors from the Fundación MEDINA acknowledge the receipt of financial support from this public-private partnership of Merck Sharp & Dohme de España S.A. with the University of Granada and Andalusian Regional Government (PIN-0474-2016).

Untargeted metabolomics (including lipidomics) is a holistic approach to biomarker discovery and mechanistic insights into disease onset and progression, and response to intervention. Each step of the analytical and statistical pipeline is crucial for the generation of high-quality, robust data. Metabolite identification remains the bottleneck in these studies; therefore, confidence in the data produced is paramount in order to maximize the biological output. Here, we outline the key steps of the metabolomics workflow and provide details on important parameters and considerations. Studies should be designed carefully to ensure appropriate statistical power and adequate controls. Subsequent sample handling and preparation should avoid the introduction of bias, which can significantly affect downstream data interpretation. It is not possible to cover the entire metabolome with a single platform; therefore, the analytical platform should reflect the biological sample under investigation and the question(s) under consideration. The large, complex datasets produced need to be pre-processed in order to extract meaningful information. Finally, the most time-consuming steps are metabolite identification, as well as metabolic pathway and network analysis. Here we discuss some widely used tools and the pitfalls of each step of the workflow, with the ultimate aim of guiding the reader towards the most efficient pipeline for their metabolomics studies.

AbstractGlobal climate changes cause water scarcity in many regions, and the sustainable use of recycled water appears crucial, especially in agriculture. However, potentially hazardous compounds such as pharmaceuticals can enter the food chain and pose severe risks. This paper aims to study the presence of selected pharmaceutical active compounds (PhACs) and their metabolites in crops grown in aeroponic conditions and evaluate the potential of PhAC plant uptake. A solvent extraction with an acidified mixture of acetonitrile and water followed by LC-HRMS was developed and validated for quantifying nine pharmaceuticals and their nine metabolites in three plants. We aimed for a robust method with a wide linear range because an extensive concentration range in different matrices was expected. The developed method proved rapid and reliable determination of selected pharmaceuticals in plants in the wide concentration range of 10 to 20,000 ng g−1 and limit of detection range 0.4 to 9.0 ng g-1. The developed method was used to study the uptake and translocation of pharmaceuticals and their metabolites in plant tissues from an aeroponic experiment at three different pH levels. Carbamazepine accumulated more in the leaves of spinach than in arugula. On the other hand, sulfamethoxazole and clindamycin evinced higher accumulation in roots than in leaves, comparable in both plants. The expected effect of pH on plants' uptake was not significant.

Ciguatera Poisoning (CP) is caused by consumption of fish or invertebrates contaminated with ciguatoxins (CTXs). Presently CP is a public concern in some temperate regions, such as Macaronesia (North-Eastern Atlantic Ocean). Toxicity analysis was performed to characterize the fish species that can accumulate CTXs and improve understanding of the ciguatera risk in this area. For that, seventeen fish specimens comprising nine species were captured from coastal waters inMadeira and Selvagens Archipelagos. Toxicity was analysed by screening CTX-like toxicity with the neuroblastoma cell-based assay (neuro-2a CBA). Afterwards, the four most toxic samples were analysed with liquid chromatography-high resolution mass spectrometry (LC-HRMS). Thirteen fish specimens presented CTX-like toxicity in their liver, but only four of these in their muscle. The liver of one specimen of Muraena augusti presented the highest CTX-like toxicity (0.270 ± 0.121 µg of CTX1B equiv·kg-1). Moreover, CTX analogues were detected with LC-HRMS, for M. augusti and Gymnothorax unicolor. The presence of three CTX analogues was identified: C-CTX1, which had been previously described in the area; dihydro-CTX2, which is reported in the area for the first time; a putative new CTX m/z 1127.6023 ([M+NH4]+) named as putative C-CTX-1109, and gambieric acid A. ; The research leading to these results has received funding from the European Union Seventh Framework Programme (FP7/2007–2013) under the ECsafeSEAFOOD project (grant agreement n° 311820). À.T. acknowledges IRTA-URV-Banco Santander for the Ph.D. grant (2016 PMF-PIPF-74). ; Peer reviewed

Carminic acid (E120) is a natural, water soluble red dye, secreted by the cochineal insect as a deterrent to predators. It is stable under conditions of indoor scattered rays, large temperature range, pH variation and oxidation-resistant [1]. This colorant is used for several uses from ink to cosmetics and foods. Regards food, it is usually added to preserve and maintain the red color of meat products during their shelf-life as well [2]. The quality of fresh meat products so may be compromised by this not admitted procedure. On this purpose, Regulation 1129/11/EC [3] prohibits the use of E120 in the fresh meat preparations, which also includes sausages . The aim of this study was to develop a method for carminic acid determination and to detect it in fresh sausages in order to guarantee to consumers a product free of illegal dye addition. In literature, almost all works detect carminic acid by liquid chromatography with UV Detector (LC-UV) system or use elaborated extraction procedures for different foods, especially for drinks [2]. In this work we developed a simple liquid extraction method with water, followed by a defatting step with hexane before analysis by liquid chromatography coupled to high resolution mass spectrometry (LC-HRMS). The LOD and LOQ of the method were 3 and 10 μg/ kg, respectively. The recovery was 99%. The other performances of the analytical method, effectiveness and robustness particularly, were evaluated following the Decision 657/2002/EC [4]. The protocol was then applied on 95 fresh sausage samples collected from different supermarkets and food suppliers, showing 23% of non-compliance with concentrations ranging from 45 to 1218 μg/ kg. The E120 evidence, not indicated on the product label of the sausages, collected directly from the different stores, suggests the importance of monitoring fresh products, safeguarding consumer health. [1] Zhang J, Li Z, Zhao J, Yang S, Bao S. Study on stability and determination methods of carminic acid extracted from dactylopiuscoccus costa, Food Science, 28: 321–326, 2007. [2] Yamjalaa K, Nainara MS, Ramisetti NR. Methods for the analysis of azo dyes employed in food industry - A review. Food Chemistry, 192: 813–824, 2016. [3] European Community. Commission Regulation (EU) No 1129/2011 amending Annex II to Regulation (EC) No 1333/2008 of the European Parliament and of the Council by establishing a Union list of food additives, Official Journal of the European Union, 295: 1-177, 2011. [4] European Community. Commision Decision 2002/657/EC concerning the performance of analytical methods and the interpretation of results, Official Journal of the European Union, 221: 8–36, 2002.

Abstract
Background
Substantial efforts have been made to monitor potentially hazardous anthropogenic contaminants in surface waters while for plant secondary metabolites (PSMs) almost no data on occurrence in the water cycle are available. These metabolites enter river waters through various pathways such as leaching, surface run-off and rain sewers or input of litter from vegetation and might add to the biological activity of the chemical mixture. To reduce this data gap, we conducted a LC–HRMS target screening in river waters from two different catchments for 150 plant metabolites which were selected from a larger database considering their expected abundance in the vegetation, their potential mobility, persistence and toxicity in the water cycle and commercial availability of standards.

Results
The screening revealed the presence of 12 out of 150 possibly toxic PSMs including coumarins (bergapten, scopoletin, fraxidin, esculetin and psoralen), a flavonoid (formononetin) and alkaloids (lycorine and narciclasine). The compounds narciclasine and lycorine were detected at concentrations up to 3ʵg/L while esculetin and fraxidin occurred at concentrations above 1ʵg/L. Nine compounds occurred at concentrations above 0.1ʵg/L, the Threshold for Toxicological Concern (TTC) for non-genotoxic and non-endocrine disrupting chemicals in drinking water.

Conclusions
Our study provides an overview of potentially biologically active PSMs in surface waters and recommends their consideration in monitoring and risk assessment of water resources. This is currently hampered by a lack of effect data including toxicity to aquatic organisms, endocrine disruption and genotoxicity and demands for involvement of these compounds in biotesting.

Although the exposure assessment of chemicals of emerging concern (CECs) has taken a decisive step forward through advances in (bio)informatics, statistics, and the development of highly sophisticated analytical instruments, the lack of standardisation and harmonisation of analytical workflows and method performance assessment for suspect and non-target screening hampers the interpretation of results, their comparability and thus, its transmission to policymakers. To date, unlike in other research fields such as forensics or food analysis, there is a lack of guidelines for non-target analysis in human risk assessment and quality assurance and quality control (QA/QC) protocols. Moreover, the majority of efforts have been focused on the development and implementation of QA/QC actions for data acquisition, data analysis and mining, largely neglecting the sample preparation necessary for determination of CECs by suspect and non-target screening methods. In this article, we propose a set of QA/QC measures that covers sampling, sample preparation and data acquisition, as an aspect of work conducted within the European Biomonitoring for Europe initiative (HBM4EU). These measures include the use of standardised terminology and the implementation of dedicated QA/QC actions in each stage of the analytical process. Moreover, a framework for the analytical performance assessment has been developed for the first time for the identification of CECs in human samples by suspect and non-target approaches. Adoption of the actions proposed here for the identification of CECs in human matrices can significantly improve the comparability of reported results and contribute to the (challenging) Exposome research field. ; The authors thank the European Union's Horizon 2020 research and innovation programme HBM4EU (Grant Agreement No. 733032) for its financial support. Dr. N. Caballero-Casero acknowledges postdoctoral fellowship from the University of Antwerp. Drs. P. Vervliet acknowledges funding through a Research Foundation Flanders project (G089016N). Drs. L. Belova acknowledges funding through a Research Foundation Flanders project (11G1821N). This work was also supported by the Exposome Centre of Excellence of the University of Antwerp (BOF grant, Antigoon database number 41222). Dr. E.J. Price acknowledges support from the Czech Operational Programme Research, Development and Education – Project Postdoc@MUNI (No. CZ.02.2.69/0.0/0.0/16_027/0008360). ; Sí

Although the exposure assessment of chemicals of emerging concern (CECs) has taken a decisive step forward through advances in (bio)informatics, statistics, and the development of highly sophisticated analytical instruments, the lack of standardisation and harmonisation of analytical workflows and method performance assessment for suspect and non-target screening hampers the interpretation of results, their comparability and thus, its transmission to policymakers. To date, unlike in other research fields such as forensics or food analysis, there is a lack of guidelines for non-target analysis in human risk assessment and quality assurance and quality control (QA/QC) protocols. Moreover, the majority of efforts have been focused on the development and implementation of QA/QC actions for data acquisition, data analysis and mining, largely neglecting the sample preparation necessary for determination of CECs by suspect and non-target screening methods. In this article, we propose a set of QA/QC measures that covers sampling, sample preparation and data acquisition, as an aspect of work conducted within the European Biomonitoring for Europe initiative (HBM4EU). These measures include the use of standardised terminology and the implementation of dedicated QA/QC actions in each stage of the analytical process. Moreover, a framework for the analytical performance assessment has been developed for the first time for the identification of CECs in human samples by suspect and non-target approaches. Adoption of the actions proposed here for the identification of CECs in human matrices can significantly improve the comparability of reported results and contribute to the (challenging) Exposome research field. ; The authors thank the European Union's Horizon 2020 research and innovation programme HBM4EU (Grant Agreement No. 733032) for its financial support. Dr. N. Caballero-Casero acknowledges postdoctoral fellowship from the University of Antwerp. Drs. P. Vervliet acknowledges funding through a Research Foundation Flanders ...

In this work the effect of chlorpyrifos exposure on metabolic profiles of zebrafish muscle was evaluated by liquid chromatography coupled to high resolution mass spectrometry. Different chemometric tools based on the selection of Regions of Interest and on Multivariate Curve-Resolution-Alternating Least Squares are proposed for the analysis of the complex data sets generated in the different exposure experiments. Analysis of Variance Simultaneous Component Analysis of changes on metabolite peak profile areas showed significant chlorpyrifos concentration and exposure time-dependent changes, clearly differentiating between exposed and non-exposed samples and between short (2 h) and long exposure times (6 h or 24 h). The changes observed in the concentrations of 50 muscle metabolites are indicative of induction of oxidative stress, of a general disruption of neurotransmitter metabolism, and of muscle exhaustion. These three effects are intimately related to the toxicity of chlorpyrifos. © 2016 Elsevier Ltd ; The research leading to these results has received funding from the European Research Council under European Union's Seven Framework Programme (FP/2007-2013)/ERC Grant Agreement n.320737. ; Peer reviewed

We are especially grateful to all the patients and their families who contributed the data that made this study possible. In addition, we thank the staff of the Clinical Research Unit of the Medical Oncology Service of the University Hospital of Jaen for their time and assistance, and the Fundacion MEDINA for their technical support. Lastly, we thank the Fundacion Bancaria Unicaja for the financial support. Jose Camacho is partly supported by the Agencia Andaluza del Conocimiento, Regional Government of Andalucia, in Spain, and ERDF (European Regional Development Fund) funds through project B-TIC-136-UGR20. ; Neoadjuvant chemotherapy (NACT) outcomes vary according to breast cancer (BC) subtype. Since pathologic complete response is one of the most important target endpoints of NACT, further investigation of NACT outcomes in BC is crucial. Thus, identifying sensitive and specific predictors of treatment response for each phenotype would enable early detection of chemoresistance and residual disease, decreasing exposures to ineffective therapies and enhancing overall survival rates. We used liquid chromatography high-resolution mass spectrometry (LC-HRMS)-based untargeted metabolomics to detect molecular changes in plasma of three different BC subtypes following the same NACT regimen, with the aim of searching for potential predictors of response. The metabolomics data set was analyzed by combining univariate and multivariate statistical strategies. By using ANOVA–simultaneous component analysis (ASCA), we were able to determine the prognostic value of potential biomarker candidates of response to NACT in the triple-negative (TN) subtype. Higher concentrations of docosahexaenoic acid and secondary bile acids were found at basal and presurgery samples, respectively, in the responders group. In addition, the glycohyocholic and glycodeoxycholic acids were able to classify TN patients according to response to treatment and overall survival with an area under the curve model > 0.77. In relation to luminal B (LB) and HER2+ subjects, it should be noted that significant differences were related to time and individual factors. Specifically, tryptophan was identified to be decreased over time in HER2+ patients, whereas LysoPE (22:6) appeared to be increased, but could not be associated with response to NACT. Therefore, the combination of untargeted-based metabolomics along with longitudinal statistical approaches may represent a very useful tool for the improvement of treatment and in administering a more personalized BC follow-up in the clinical practice. ; Fundacion MEDINA ; Fundacion Bancaria Unicaja ; Agencia Andaluza del Conocimiento, Regional Government of Andalucia ; European Commission B-TIC-136-UGR20