HAL ENVT (Ecole Nationale Vétérinaire de Toulouse)
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Integrating NMR and MS for Improved Metabolomic Analysis: From Methodologies to Applications
No full textInternational audienceMetabolomics, the comprehensive analysis of low-molecular-weight metabolites (typically below 1500 DA) in biological systems, relies heavily on mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy. Each technique has inherent strengths and weaknesses. MS offers high sensitivity and is commonly coupled with chromatography to analyze complex matrices, yet it is destructive, has limited reproducibility, and provides limited structural information. NMR, while less sensitive, is non-destructive and enables structural elucidation and precise quantification. Recent studies increasingly employ data fusion (DF) strategies to combine the complementary information from NMR and MS, aiming to enhance metabolomic analyses. This review summarizes DF methodologies using NMR and MS data in metabolomics studies over the past decade. A comprehensive search of SciFinder, Scopus, and Clarivate Web of Science databases was conducted to analyze fusion techniques, methods, and statistical models. The review emphasizes the growing importance of DF in metabolomics, showing its capacity to provide a more comprehensive view of biochemical processes across diverse biological systems, including clinical, plant, and food matrices
Miniaturized workflow for the analysis of oxilipins: from extraction to analysis
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Validation de l’automate catalyst one ® dans l’analyse des épanchements canins et félins au sein du laboratoire des urgences de l’École Nationale Vétérinaire de Toulouse : étude de la répétabilité à court terme
No full textThis project focused on evaluating the validity of the Catalyst One®, a blood biochemistry analyzer commonly used in veterinary practice, for the analysis of canine and feline effusions. A simplified method validation procedure was carried out in accordance with ASVCP recommendations. Short-term repeatability for the selected biochemical variables was generally acceptable for the types of effusions analyzed. However, it could not be assessed for analyte concentrations falling outside the quantification ranges of the analyzer. Moreover, as the reference standards used were those applicable to serum or plasma analyses, the results must be interpreted with caution, considering the expected differences in performance when applied to effusion samples. Lastly, these findings should be interpreted in conjunction with the results from other sections of this study in order to assess the overall validity of the method.Ce projet consistait en l’étude de la validité du Catalyst One ®, analyseur de biochimie sanguine communément utilisé en pratique vétérinaire, dans le cadre de l’analyse des épanchements canins et félins. Une procédure simplifiée de validation de méthode a été menée selon les recommandations de l’ASVCP. La répétabilité à court terme pour les variables biochimiques retenues était souvent acceptable pour les types d’épanchements analysés. Cependant, celle-ci n’était pas évaluable pour des concentrations d'analytes extérieures aux intervalles de quantification de l’automate. De plus, les valeurs comparatives utilisées étant celles relatives aux analyses de sérum ou plasma, il convient de nuancer les résultats en regard des différences de performances attendues dans le cas des épanchements. Enfin, ces résultats sont également à mettre en perspective avec ceux des autres parties de cette étude afin de discuter de la validation de la méthode
New effective and less painful high frequency electrochemotherapy protocols: From optimization on 3D models to pilot study on veterinary patients
No full textInternational audienceElectroporation, a physical method that permeabilizes cell membranes, is increasingly used in cancer treatment. By enhancing the uptake of hydrophilic antitumor drugs, it boosts their cytotoxic effects and has proven effective in both human and veterinary medicine through electrochemotherapy. However, this treatment requires loco-regional or even general anesthesia, as electrical pulses cause muscle contractions and pain. Several clinical studies have demonstrated that application of high frequency pulses (above 5,000 Hz) and short pulse duration (under 11 µs) causes much less discomfort to patients. In order to reduce the pain associated with contractions while maintaining the effectiveness of the treatment, we have developed new protocols using a high-frequency generator that delivers electric field pulses at a pulse repetition rate up to 2 MHz, associated to a multipolar electrode. In vitro tests on colorectal cancer cells were performed to assess the efficiency of cisplatin and bleomycin in inducing cell death. The efficiency obtained after one single treatment on both cell suspensions and on 3D multicellular spheroid models were similar to the ones obtained using ESOPE (European standard operating procedures for electrochemotherapy) protocol, which is currently used in clinics. In addition, as tumor cells die in an immunogenic cell death (ICD) mode and can release danger associated molecular patterns (DAMPs), major hallmarks of ICD were evaluated following the treatment by quantifying the apoptotic cell death, caspases 3/7 activation and key DAMPs. Subsequently, pilot studies on small number of conscious cats and horses under mild sedation confirmed that these protocols did not cause any noticeable muscle contractions and resulted in either partial or complete responses. New high-frequency electroporation protocols, described herein, show great promise in shifting electrochemotherapy into an effective and painless cancer treatment.</div
Genomic and morphometric evidence for Austronesian-mediated pig translocation in the Pacific
No full textInternational audienceHuman mediated translocation of non-native pig species (genus Sus) to the islands ofWallacea and Oceania has significantly altered local ecosystems. To investigate the timing and trajectory of these introductions, we conducted both genomic analyses of 576 pig nuclear genomes and a geometric morphometric analysis of 714 modern and ancient dental remains. Our analyses demonstrate that feral and domestic pigs in Wallacea and Oceania possess diverse ancestries resulting from the introduction of multiple, sequential pig populations followed by 20 gene flow. Despite the variability in their genomic ancestry these pigs all possess a distinct tooth morphology, and a genetic link to the Chinese domestic pig populations that accompanied the dispersal of Austronesian language speakers ~4,000-3,000 years ago via Taiwan and the Philippines
Production et commercialisation du miel : actualisation des connaissances et du cadre réglementaire
No full textHoney, a natural product made from nectar transformed by bees, contains a diverse array of sugars, enzymes, and bioactive compounds that provide unique nutritional and therapeutic properties. However, honey is vulnerable to various contaminants. These contaminants pose significant challenges to honey quality and safety, affecting both human health and bee populations. Analytical methods enable the detection and monitoring of these substances to ensure product compliance and consumer protection. Strict European and international regulations govern the honey sector, facilitating traceability, quality assurance, and consumer confidence. Effective management of these issues is vital to safeguard this valuable natural product and its ecological and economic roles.Le miel, produit naturel issu du nectar transformé par les abeilles, rassemble une grande diversité de sucres, d’enzymes et de composants bioactifs qui lui confèrent des propriétés nutritionnelles et thérapeutiques uniques. Cependant, ce produit naturel est exposé à diverses contaminations potentielles. Ces contaminations affectent la qualité et la sécurité du miel, posant des enjeux importants liés à la santé humaine et à la préservation des colonies d’abeilles. Des méthodes analytiques permettent la détection et le contrôle de ces substances, fournissant des garanties indispensables pour la conformité et la sécurité des produits commercialisés. Un cadre réglementaire européen et international strict encadre cette filière afin d’assurer la traçabilité, la qualité, et la confiance des consommateurs dans ce produit précieux
Cellular Heterogeneity Analysis: An Innovative Approach to Single-Cell Metabolomics
No full textInternational audienc
Identification method for unknown disturbances in growing pigs: a case study
No full textInternational audienceThere are two ways to approach pig’s robustness based on longitudinal data. One is to analyze the pattern of these data without previous knowledge on events that may have affected the animals, such as LSR. Another is to search for indicators of disturbances in the data and then perform assessment of robustness with that prior knowledge. Axiom boar testing station was equipped with electronic feeders (EFS) allowing daily monitoring of feed intake (DFI) and body weight (BW). For on-farm monitoring, alarms indicators, i.e. detection of abnormal events, werecalculated from deviation of DFI or BW compared to the previous days for individual and pen level. In the case of low impact at individual level or missing human recording, alarms may not be emitted. The method UpDown (UD) was developed to identify disturbances and to estimate their intensity and duration with a high specificity at batch and/or pen level. UD was developed using simulated data and was validated using real data from a test farm. This study aimed at comparing the overlap of events detected by UD with alarm indicators recorded at Axiom boar testing station. Individual weight and individual feed intake from EFS of 7456 growers bred between 2019 and 2024 in 74 batches were analyzed with UD R-package with at least 4 pens per batches and 7 animals per pen. To identify disturbance with high technical impact, threshold parameter of UD was set at 95%. 5 batches were detected as disturbed for DFI and 0 batches but 3 pens detected for BW. As expected for a genetic farm with high sanitary status, a few groups appears as disturbed with 95% of individual affected. All groups detected by UD approach had alarms available, confirming a disturbance. UD detected global disturbances and their duration when alarm information caught only the most important variation happening usually on one day. UD is useful also to detect disturbance at posteriori independently of events registration. Such information could be used to interpret results from LSR from a more practical perspective
Développement de méthodologies inspirées de la métabolomique pour l'attribution chimique d'un pesticideorganophosphoré (chlorpyrifos) issu de plusieurs sources de synthèse.
No full textIn events involving a toxic chemical agent, it is crucial to identify the agent's product on history. However, low concentration, sample aging, and complex matrices make this task particularly challenging. Chemical signature-based profiling enables the linkage of a chemical agent to its origin, providing valuable support in forensic investigations. This work developed and tested analytical and chemometrics methods, adapted from mass spectrometry-based metabolomics, to characterize these compounds and determine their origin. A proof of concept was carried out on chlorpyrifos, an organophosphorus compound model, synthesized using different protocols. The method was first developed on laboratory samples and later validated, two years afterward, on unknown blind samples. Finaly, the robustness of the approach was assessed using environmental samples (water, soil) spiked with raw syntheses of chlorpyrifos at varying concentrations, applying statistical models to predict the synthetic routes. The combination of multidimensional analyses (LC-HRMS, MS/MS, ion mobility) and chemometrics proved effective in addressing analytical challenges in the context of defense.Lors d’événements impliquant un agent chimique toxique, il est crucial d’identifier l’historique de cet agent. Cependant, la faible concentration, le vieillissement des échantillons et la complexité des matrices rendent cette tâche difficile. Le profilage basé sur les signatures chimiques a pour but la détermination d’informations permettant d’aider à relier un agent chimique à son origine. L’analyse des impuretés peut renseigner sur la synthèse, le lot de production ou bien de façon plus générale sur l’origine et l’historique de l’agent. Lors de ce travail, une preuve de concept impliquant des méthodes analytiques et de chimiométrie issues de la métabolomique par spectrométrie de masse a été développée et testée pour la caractérisation et l’inférence des sources d’un composé organophosphoré (le chlorpyrifos) produit selon plusieurs voies de synthèse. La méthode a été développée sur des échantillons connus puis validée deux ans plus tard sur des échantillons inconnus analysés à l’aveugle. Enfin, la robustesse de la méthode a été testée en analysant des matrices environnementales (eaux, sols) contaminées à différentes concentrations, avec des modèles statistiques pour prédire les voies de synthèse. La combinaison d’analyses multidimensionnelles (LC-HRMS, MS/MS, mobilité ionique) et de chimiométrie a montré son efficacité pour relever les défis analytiques posés
