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Environmental pollutants used as plasticizers in food packaging and in thousands of everyday products have become harmful for their impact on human health. Among them, phthalates, recognized as emerging endocrine disruptors (EDs) can induce toxic effects leading to different health disorders. Only few studies evaluated the effects of di-n-hexyl phthalate (DnHP) in in vivo models and no studies have been conducted to investigate the effect of DnHP on the endocannabinoid system (ECS), one of the majors signaling pathways involved in the microbiota-gut-brain axis. Due to the current relevance of probiotic bacteria as beneficial dietary interventions, the present study was aimed at evaluating the potential neuroprotective impact of daily administration of Lactiplantibacillus (Lpb.) plantarum IMC513 on zebrafish adults exposed to DnHP, with a focus on ECS modulation. Gene expression analysis revealed a promising protective role of probiotic through the restoration of ECS genes expression to the control level, in the brain of zebrafish daily exposed to DnHP. In addition, the levels of main endocannabinoids were also modulated. These findings confirm the potential ability of probiotics to interact at central level by modulating the ECS, suggesting the use of probiotics as innovative dietary strategy to counteract alterations by EDs exposure.
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Endocannabinoides , Probióticos , Pez Cebra , Animales , Probióticos/administración & dosificación , Probióticos/farmacología , Endocannabinoides/metabolismo , Ácidos Ftálicos/toxicidad , Encéfalo/metabolismo , Encéfalo/efectos de los fármacosRESUMEN
The demand for plant-based protein sources in the food industry has significantly increased in recent years, leading to the introduction of legume-based products as meat substitutes. However, concerns regarding food quality have emerged, particularly related to the presence of mycotoxins. This study addresses the need for the sensitive detection of phomopsins (PHOs), a class of peptide-based toxins. A selective extraction method using molecularly imprinted polymer (MIP) coupled with ultra-high performance liquid chromatography and tandem mass spectrometry (UHPLC-MS/MS) was focused on the most toxic Phomopsin A (PHO-A). A rapid ultrasonochemical synthesis of MIP (5 min) was proposed and its performance was optimized in response to various factors, including the choice of dummy template and the selection of the monomer. The methacrylic acid-vinyl pyridine (MAA-VP) MIP exhibited high selectivity and affinity for PHO-A. The method was tested in lupin samples and the validation, according to SANTE/11312/2021 international guidelines, gave excellent recovery (80-90 %), low matrix effects, and high accuracy and precision. Real samples analysis confirmed the presence of PHO-A in artificially fungal inoculated lupins, with levels ranging from 0.377 to 0.576 mg kg-1. In order to identify further PHOs, a semi-untargeted approach using multiple reaction monitoring-information dependent acquisition-enhanced product ion (MRM-IDA-EPI) was developed. PHO-B, PHO-D, PHO-E and PHO-P, rarely previously reported in lupin matrix, were tentatively identified. This study accounts for the effectiveness of MIP-based extraction coupled with UHPLC-triple quadrupole with linear ionic trap-MS/MS (UHPLC-QqQ-LIT-MS/MS) for quantification of PHO-A and putative detection of other PHOs, offering a promising method for investigating this class of toxins in food.
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Lupinus , Polímeros Impresos Molecularmente , Espectrometría de Masas en Tándem , Espectrometría de Masas en Tándem/métodos , Cromatografía Líquida de Alta Presión/métodos , Polímeros Impresos Molecularmente/química , Lupinus/química , Contaminación de Alimentos/análisis , Micotoxinas/análisis , Micotoxinas/química , Impresión Molecular , OligopéptidosRESUMEN
Neuroinflammation is a hallmark of several neurodegenerative disorders that has been extensively studied in recent years. Microglia, the primary immune cells of the central nervous system (CNS), are key players in this physiological process, demonstrating a remarkable adaptability in responding to various stimuli in the eye and the brain. Within the complex network of neuroinflammatory signals, the fatty acid N-ethanolamines, in particular N-arachidonylethanolamine (anandamide, AEA), emerged as crucial regulators of microglial activity under both physiological and pathological states. In this study, we interrogated for the first time the impact of the signaling of these bioactive lipids on microglial cell responses to a sub-lethal acute UVB radiation, a physical stressor responsible of microglia reactivity in either the retina or the brain. To this end, we developed an in vitro model using mouse microglial BV-2 cells. Upon 24 h of UVB exposure, BV-2 cells showed elevated oxidative stress markers and, cyclooxygenase (COX-2) expression, enhanced phagocytic and chemotactic activities, along with an altered immune profiling. Notably, UVB exposure led to a selective increase in expression and activity of fatty acid amide hydrolase (FAAH), the main enzyme responsible for degradation of fatty acid ethanolamides. Pharmacological FAAH inhibition via URB597 counteracted the effects of UVB exposure, decreasing tumor necrosis factor α (TNF-α) and nitric oxide (NO) release and reverting reactive oxidative species (ROS), interleukin-1ß (IL-1ß), and interleukin-10 (IL-10) levels to the control levels. Our findings support the potential of enhanced fatty acid amide signaling in mitigating UVB-induced cellular damage, paving the way to further exploration of these lipids in light-induced immune responses.
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Amidohidrolasas , Microglía , Rayos Ultravioleta , Microglía/metabolismo , Microglía/efectos de los fármacos , Microglía/efectos de la radiación , Animales , Ratones , Amidohidrolasas/metabolismo , Amidohidrolasas/antagonistas & inhibidores , Rayos Ultravioleta/efectos adversos , Línea Celular , Carbamatos/farmacología , Benzamidas/farmacología , Estrés Oxidativo/efectos de los fármacos , Endocannabinoides/metabolismo , Ciclooxigenasa 2/metabolismo , Ciclooxigenasa 2/genética , Ácidos Araquidónicos/metabolismo , Ácidos Araquidónicos/farmacología , Interleucina-10/metabolismo , Alcamidas PoliinsaturadasRESUMEN
BACKGROUND: Novel psychoactive substances (NPS) are a group of substances, mainly of synthetic origin, characterized by toxicological properties extremely dangerous. The main difficulty in recognizing NPS in seizures and biological samples lies in their dynamic nature, related to the continuous synthesis and introduction on the market of new drugs, often with very similar structures to existing ones. The aim of this study was the creation of a robust and versatile method for the analysis of traditional drugs and NPS in different matrices. RESULTS: Both target analysis and suspect screening methodologies were developed. The strategy used for suspect screening allowed to collect data through a scheduled multi reaction monitoring (sMRM) survey which triggered the collection of enhanced product ion (EPI) spectra when a compound met information dependent acquisition (IDA) criteria. The retention time of the different drugs, which was crucial to define the sMRM survey scan parameters, was predicted with a Quantitative Structure Retention (Chromatographic) Relationship (QSRR) model by Multiple Linear Regression. The model was validated through the evaluation of training set predictions, an external validation set and a leave-one out strategy; the results showed that the method fit for its purpose. The target method was validated in oral fluid as a testing matrix, with excellent results in term of recovery, accuracy, precision and matrix effect. Finally, the performances of the suspect method were evaluated by analysing a mixture containing 8 reference standards not included in the initial dataset, as well as seizures and real oral fluid samples. Four NPS were putatively identified in the analysed samples. SIGNIFICANCE: The advantage of the proposed approach is the possibility of quantifying 65 classical drugs of abuse and NPS and, at the same time, detect and putatively identify 146 additional drugs in one single LC-MS/MS run. This is an innovative strategy for multi analyte detection and enables detection of low concentrations of drugs in complex biological matrices such as oral fluid. Considering the highly dynamic drug market, a strength of this strategy is that the analytical method can be kept up to date through the addition of new compounds based on the last drug monitoring bodies alerts without the need of authentic standards.
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Cromatografía Líquida con Espectrometría de Masas , Espectrometría de Masas en Tándem , Humanos , Cromatografía Liquida , Monitoreo de Drogas , ConvulsionesRESUMEN
Novel psychoactive substances (NPS) represent a broad class of drugs new to the illicit market that often allow passing drug-screening tests. They are characterized by a variety of structures, rapid transience on the drug scene and mostly unknown metabolic profiles, thus creating an ever-changing scenario with evolving analytical targets. The present study aims at developing an indirect screening strategy for NPS monitoring, and specifically for new synthetic opioids (NSOs), based on assessing changes in endogenous urinary metabolite levels as a consequence of the systemic response following their intake. The experimental design involved in-vivo mice models: 16 animals of both sex received a single administration of morphine or fentanyl. Urine was collected before and after administration at different time points; the samples were then analysed with an untargeted metabolomics LC-HRMS workflow. According to our results, the intake of opioids resulted in an elevated energy demand, that was more pronounced on male animals, as evidenced by the increase in medium and long chain acylcarnitines levels. It was also shown that opioid administration disrupted the pathways related to catecholamines biosynthesis. The observed alterations were common to both morphine and fentanyl: this evidence indicate that they are not related to the chemical structure of the drug, but rather on the drug class. The proposed strategy may reinforce existing NPS screening approaches, by identifying indirect markers of drug assumption.
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Analgésicos Opioides , Fentanilo , Metabolómica , Morfina , Animales , Masculino , Femenino , Ratones , Metabolómica/métodos , Analgésicos Opioides/orina , Fentanilo/análogos & derivados , Fentanilo/orina , Fentanilo/metabolismo , Cromatografía Líquida de Alta Presión/métodos , Morfina/orina , Psicotrópicos/orina , Espectrometría de Masas/métodos , Metaboloma/efectos de los fármacosRESUMEN
Space-related stressors such as microgravity are associated with cellular and molecular alterations of the immune and inflammatory homeostasis that have been linked to the disorders that astronauts suffer from during their missions. Most of the research of the past 30 years has consistently established that innate adaptive immune cells represent a target of microgravity, which leads to their defective or dysfunctional activation, as well as to an altered ability to produce soluble mediators-e.g., cytokines/chemokines and bioactive lipids-that altogether control tissue homeostasis. Bioactive lipids include a vast array of endogenous molecules of immune origin that control the induction, intensity and outcome of the inflammatory events. However, none of the papers published so far focus on a newly characterized class of lipid mediators called specialized pro-resolving mediators (SPMs), which orchestrate the "resolution of inflammation"-i.e., the active control and confinement of the inflammatory torrent mostly driven by eicosanoids. SPMs are emerging as crucial players in those processes that avoid acute inflammation to degenerate into a chronic event. Given that SPMs, along with their metabolism and signaling, are being increasingly linked to many inflammatory disorders, their study seems of the outmost importance in the research of pathological processes involved in space-related diseases, also with the perspective of developing therapeutic countermeasures. Here, we show that microgravity, simulated in the rotary cell culture system (RCCS) developed by NASA, rearranges SPM receptors both at the gene and protein level, in human monocytes but not in lymphocytes. Moreover, RCCS treatment reduces the biosynthesis of a prominent SPM like resolvin (Rv) D1. These findings strongly suggest that not only microgravity can impair the functioning of immune cells at the level of bioactive lipids directly involved in proper inflammation, but it does so in a cell-specific manner, possibly perturbing immune homeostasis with monocytes being primary targets.
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Monocitos , Ingravidez , Humanos , Homeostasis , Citocinas , InflamaciónRESUMEN
BACKGROUND: Salt has been identified as an elicitor that can increase the accumulation of phytochemicals in seedlings during the germination process. However, the salinity level required to maximize the yield of phytochemicals, particularly phenolic compounds, needs further investigation for several plant species. To address this issue, we imposed increasing levels of salinity (NaCl solutions) on the sprouting substrate of Triticum durum (var. Platone) grains, at concentrations of 0, 50, 100, 150, 200, 250, and 300 mM (0_S, 50_S, 100_S, 150_S, 200_S, 250_S, and 300_S, respectively). RESULTS: The highest NaCl doses (250_S and 300_S) significantly impacted germination performance and were excluded from further analysis. The seedlings harvested at 8 days after sowing exhibited different growth stages depending on the salinity level: wheatgrass for 0_S, early wheatgrass for 50_S, intermediate between sprout and wheatgrass for 100_S, sprout for 150_S, and very early sprout for 200_S. Furthermore, salinity induced the concentration of phenolic compounds (PhCs) in the seedlings' tissues (i.e., both roots and shoots) in a salinity-dependent manner. The highest values were observed at 200_S, with an increase of 187% of the total investigated PhCs in comparison with 0_S, averaged over shoots and roots. In particular, in 200_S, the accumulation of phenolic acids was up to fourfold higher in roots, and that of flavonoids was up to twofold higher in shoots. CONCLUSION: Our findings suggest that the use of 200 mM NaCl applied to the sprouting substrate is excessive for producing edible sprouts but may be suitable for phytochemical extraction purposes. © 2023 Society of Chemical Industry.
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Plantones , Triticum , Triticum/química , Cloruro de Sodio/análisis , Antioxidantes/química , Fenoles/química , Fitoquímicos/química , SalinidadRESUMEN
Introduction: The analysis of organic residue in ancient vessels to investigate early-age civilization habits is an important archeological application that needs advanced analytical methods. However, these procedures should meet inherent requisites such as low sampling invasiveness and high sensitivity for trace analysis. This study deals with the development of advanced analytical methods for the detection of opium alkaloids in ceramic vessels and its first application to the study of Daunian pots dating back to the VIII-IV sec BC. Methods: All the stages of the analytical procedure, from sampling to analysis, were carefully optimized. Concerning sampling, the traditional scraping approach was compared with a swabbing strategy which permitted minimizing sample encroachment. Extraction was based on pressurized liquid extraction or ultrasound-assisted liquid extraction, followed by dispersive liquid-liquid microextraction, which allowed concentration enrichment. On the other hand, a UHPLC-MS/MS method was specifically developed and validated to obtain reliable data. Some Daunian pots, belonging to the Ceci-Macrini private archeological collection, were selected for sample withdrawal as their iconography could suggest opium usage. Results: Several of the analyzed samples resulted positive to thebaine and less frequently to morphine and codeine; furthermore, 70% of the analyzed items tested positive for at least one opium alkaloid. Positive findings were common to all the samples collected in the pots, suggesting that scraping and swabbing provided comparable results and validating this unusual sampling strategy. All samples were additionally analyzed by UHPLC-HRMS to further improve the confidence level of the identified compounds. The obtained results shed new light on the hypothesis of opium usage by the ancient Daunian civilization. Furthermore, this study provided suitable analytical tools for further investigations on the same topic, with a good level of confidence in the quality of the results.
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To date, it is still not possible to obtain exhaustive information about organic materials in cultural heritage without sampling. Nonetheless, when studying unique objects with invaluable artistic or historical significance, preserving their integrity is a priority. In particular, organic dye identification is of significant interest for history and conservation research, but it is still hindered by analytes' low concentration and poor fastness. In this work, a minimally invasive approach for dye identification is presented. The procedure is designed to accompany noninvasive analyses of inorganic substances for comprehensive studies of complex cultural heritage matrices, in compliance with their soundness. Liquid extraction of madder, turmeric, and indigo dyes was performed directly from paint layers and textiles. The extraction was supported by hydrogels, which themselves can undergo multitechnique analyses in the place of samples. After extraction, Ag colloid pastes were applied on the gels for SERS analyses, allowing for the identification of the three dyes. For the HPLC-MS/MS analyses, re-extraction of the dyes was followed by a clean-up step that was successfully applied on madder and turmeric. The colour change perceptivity after extraction was measured with colorimetry. The results showed ΔE values mostly below the upper limit of rigorous colour change, confirming the gentleness of the procedure.
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The introduction of synthetic dyes completely changed the industrial production and use of colorants for art materials. From the synthesis of the first synthetic dye, mauveine, in 1856 until today, artists have enjoyed a wider range of colors and selection of chemical properties than was ever available before. However, the introduction of synthetic dyes introduced a wider variety and increased the complexity of the chemical structures of marketed dyes. This work looks towards the analysis of synthetically dyed objects in heritage collections, applying an extraction protocol based on the use of ammonia, which is considered favorable for natural anthraquinone dyes but has never before been applied to acid synthetic dyes. This work also presents an innovative cleanup step based on the use of an ion pair dispersive liquid-liquid microextraction for the purification and preconcentration of historical synthetic dyes before analysis. This approach was adapted from food science analysis and is applied to synthetic dyes in heritage science for the first time in this paper. The results showed adequate recovery of analytes and allowed for the ammonia-based extraction method to be applied successfully to 15 samples of suspected azo dyes from the Azienda Coloranti Nazionali e Affini (ACNA) synthetic dye collection, identified through untargeted HPLC-HRMS analyses.
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The aim of the present work was to evaluate if the use of grape pomace (GP) in the feeding of dairy ewes can improve the content of phenolic compounds (PCs) in the milk and affect the anti-inflammatory and antioxidative status of the milk. For this purpose, 46 ewes were randomly assigned to two groups of 23 animals each: a control group (Ctrl) that received a standard diet and an experimental group (GP+), whose diet was been formulated with 10% GP on a dry matter (DM) basis. At the end of the 60 days of the trial, from 10 ewes selected randomly from each group, individual milk samples were collected and analyzed for the identification and the quantification of phenolic compounds through an ultra-high-performance liquid chromatography system, and milk anti-inflammatory and antioxidative status were evaluated by enzyme-linked immunosorbent assay, determining the activity of GPx and CAT and the levels of IL-1 and TNFα. In addition, gelatinolytic activity of Type IV collagenases (MMP-2/MMP-9) was evaluated by the fluorometric method and zymographic approach. The results obtained showed that the diet with GP affects the phenolic profile of milk, inducing milk enrichment of phenolic compounds without, however, having a significant impact on milk antioxidant and inflammatory status. However, a lower activity of MMP-9 was found in GP+ milk. The use of the molecular docking approach showed the ability of luteolin to approach the catalytic pocket of the enzyme, interfering with the recruitment of the substrate, and therefore, slowing down their hydrolytic activity.
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Leche , Vitis , Animales , Femenino , Ovinos , Leche/química , Antioxidantes/análisis , Vitis/química , Metaloproteinasa 9 de la Matriz/análisis , Lactancia , Simulación del Acoplamiento Molecular , Fenoles/análisisRESUMEN
Wheat is critical for food security, and is challenged by biotic stresses, chiefly aphids and the viruses they transmit. The objective of this study was to determine whether aphids feeding on wheat could trigger a defensive plant reaction to oxidative stress that involved plant oxylipins. Plants were grown in chambers with a factorial combination of two nitrogen rates (100% N vs. 20% N in Hoagland solution), and two concentrations of CO2 (400 vs. 700 ppm). The seedlings were challenged with Rhopalosiphum padi or Sitobion avenae for 8 h. Wheat leaves produced phytoprostanes (PhytoPs) of the F1 series, and three types of phytofurans (PhytoFs): ent-16(RS)-13-epi-ST-Δ14-9-PhytoF, ent-16(RS)-9-epi-ST-Δ14-10-PhytoF and ent-9(RS)-12-epi-ST-Δ10-13-PhytoF. The oxylipin levels varied with aphids, but not with other experimental sources of variation. Both Rhopalosiphum padi and Sitobion avenae reduced the concentrations of ent-16(RS)-13-epi-ST-Δ14-9-PhytoF and ent-16(RS)-9-epi-ST-Δ14-10-PhytoF in relation to controls, but had little or no effect on PhytoPs. Our results are consistent with aphids affecting the levels of PUFAs (oxylipin precursors), which decreased the levels of PhytoFs in wheat leaves. Therefore, PhytoFs could be postulated as an early indicator of aphid hosting for this plant species. This is the first report on the quantification of non-enzymatic PhytoFs and PhytoPs in wheat leaves in response to aphids.
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Áfidos , Oxilipinas , Animales , Áfidos/fisiología , Triticum , Dióxido de Carbono , Hojas de la PlantaRESUMEN
Dysfunctional phenotype of microglia, the primary brain immune cells, may aggravate Alzheimer's disease (AD) pathogenesis by releasing proinflammatory factors, such as nitric oxide (NO). The endocannabinoids N-arachidonoylethanolamine (AEA) and 2-arachidonoylglycerol (2-AG) are bioactive lipids increasingly recognised for their essential roles in regulating microglial activity both under normal and AD-driven pathological conditions. To investigate the possible impact of chronic exposure to ß-amyloid peptides (Aß) on the microglial endocannabinoid signalling, we characterised the functional expression of the endocannabinoid system on neonatal microglia isolated from wild-type and Tg2576 mice, an AD-like model, which overexpresses Aß peptides in the developing brain. We found that Aß-exposed microglia produced 2-fold more 2-AG than normal microglia. Accordingly, the expression levels of diacylglycerol lipase-α (DAGLα) and monoacylglycerol lipase (MAGL), the main enzymes responsible for synthesising and hydrolysing 2-AG, respectively, were consistently modified in Tg2576 microglia. Furthermore, compared to wild-type cells, transgenic microglia basally showed increased expression of the cannabinoid 2 receptor, typically upregulated in an activated proinflammatory phenotype. Indeed, following inflammatory stimulus, Aß-exposed microglia displayed an enhanced production of NO, which was abolished by pharmacological inhibition of DAGLα. These findings suggested that exposure to Aß polarises microglial cells towards a pro-AD phenotype, possibly by enhancing 2-AG signalling.
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Enfermedad de Alzheimer , Microglía , Ratones , Animales , Microglía/metabolismo , Endocannabinoides/metabolismo , Transducción de Señal/fisiología , Enfermedad de Alzheimer/metabolismo , Péptidos beta-Amiloides/metabolismo , Receptores de Cannabinoides/metabolismo , Ratones TransgénicosRESUMEN
The present study encompasses the development of a fast and reliable analytical method to quantify the main endocannabinoids and some of their conjugated congeners, particularly N-arachidonoyl amino acids, in brain tissue. Samples were homogenized and a micro solid phase extraction (µSPE) procedure was developed for brain homogenate clean-up. Miniaturized SPE was selected as it allowed to work with reduced sample amounts, while maintaining high sensitivity; this last feature was mandatory due to the low concentration of endocannabinoids in biological matrices that makes their determination a challenging analytical task. UHPLC-MS/MS was used for the analysis as it provided a great sensitivity, especially for conjugated forms that were detected by negative ionization. Polarity switching was applied during the run; low limits of quantification were between 0.003 ng g-1 and 0.5 ng g-1. This method provided also low matrix effect (lower than 30%) and good extraction recoveries in the brain. To the best of our knowledge, this is the first time that µSPE is applied on this matrix for this class of compounds. The method was validated according to international guidelines, and then tested on real cerebellum samples from mice, which were sub-chronically treated with URB597, a well-known inhibitor of the fatty acid amide hydrolase.
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Endocannabinoides , Espectrometría de Masas en Tándem , Animales , Ratones , Cromatografía Líquida de Alta Presión/métodos , Endocannabinoides/química , Espectrometría de Masas en Tándem/métodos , Extracción en Fase Sólida/métodos , EncéfaloRESUMEN
Lupin alkaloids (LAs) represent a class of toxic secondary metabolites in plants, in particular in Lupinus spp.; they are produced as a defense mechanism due to their strong bitter taste and are very dangerous for human and animals. In this work, a sensitive and reliable high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) analytical method for the identification and quantification of thirteen lupin alkaloids was developed and validated according to FDA guidelines. Efficient extraction and clean-up steps, carried out by solid-phase extraction, were finely tuned on the basis of the characteristics of the analytes and lupin samples, providing good selectivity with minimized matrix interference. The effectiveness of the method was proven by the satisfactory recovery values obtained for most of the analytes and a matrix effect ≤23% for all tested levels. In addition, a sensitive and reliable determination of the target compounds was obtained; LOQs were between 1 and 25 µg Kg-1, i.e., below the requested maximum levels (<200 mg Kg-1). The method was applied to evaluate the LAs profile in different batches of raw L. albus L. samples, varying in size and across farming treatments.
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Alcaloides , Lupinus , Animales , Humanos , Cromatografía Líquida de Alta Presión/métodos , Espectrometría de Masas en Tándem/métodos , Lupinus/química , Alcaloides/química , Extracción en Fase SólidaRESUMEN
Aflatoxins (AFs) are fungi secondary metabolites produced by the Aspergillus family. These compounds can enter the food chain through food contamination, representing a risk to human health. Commercial immunoaffinity columns are widely used for the extraction and cleanup of AFs from food samples; however, their high cost and large solvent consumption create a need for alternative strategies. In this work, an alternative strategy for producing molecularly imprinted polymers (MIPs) was proposed to extract aflatoxins AFB1, AFB2, AFG1, and AFG2 from complex food samples, using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The MIPs were synthesized via a low-cost and rapid (5 min) sonochemical free-radical polymerization, using 1-hydroxy-2-naphthoic acid as a dummy template. MIPs-based solid phase extraction performance was tested on 17 dietary supplements (vegetables, fruits, and cereals), obtaining appreciable recovery rates (65-90%) and good reproducibility (RSD ≤ 6%, n = 3); the selectivity towards other mycotoxins was proved and the data obtained compared with commercial immunoaffinity columns. The proposed strategy can be considered an alternative affordable approach to the classical immunoaffinity columns, since it is more selective and better performing.
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Aflatoxinas , Contaminación de Alimentos , Aflatoxinas/análisis , Cromatografía Líquida de Alta Presión/métodos , Cromatografía Liquida/métodos , Contaminación de Alimentos/análisis , Polímeros Impresos Molecularmente/análisis , Reproducibilidad de los Resultados , Extracción en Fase Sólida , Espectrometría de Masas en Tándem/métodosRESUMEN
In this work, the chemical composition and the antioxidant evaluation of the inflorescences from 12 Cannabis sativa L. monoecious cultivars (Carmagnola Lemon CL, Ferimon F, Gran Sasso Kush GSK, Antal A, Carmagnola C, Kompolti K, Futura 75 F75, Villanova V, Tiborzallasi T, Finola FL, Kc Virtus KV and Pineapple P) cultivated at the same condition, were investigated. GC-MS analysis was carried out to evaluate the volatile fraction, while HPLC-MS/MS was used for cannabinoids and polyphenolic compounds. The evaluation of antioxidant activity was carried out using ABTS*+, Trolox equivalence antioxidant capacity (TEAC), ferric reducing antioxidant property (FRAP) and 2,2-diphenyl-1-picrylhydrazyl (DPPH*) assays in vitro. The obtained data, demonstrated that each cultivar has a characteristic chemical profile, with highest antioxidant capacity for CL, F75, GSK and F. Based on the in vitro antioxidant activity the plant extracts can be considered as promising candidates for different applications in food field.
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Cannabinoides , Cannabis , Cannabis/química , Antioxidantes/análisis , Espectrometría de Masas en Tándem , Cannabinoides/química , Extractos Vegetales/análisisRESUMEN
LC-MS/MS is a powerful analytical technique that provides unequivocal identification and reliable quantification of the analytes, using Selected Reaction Monitoring or Multi Reaction Monitoring acquisition mode.Anandamide (N-arachidonoylethanolamine, AEA) and 2-Arachidonoylglycerol (2-AG) are the most abundant endocannabinoids (eCBs), which play a major role in a wide variety of physiological and pathological processes. Analysis of those compounds by means of LC-MS/MS allows the detection of very low concentrations in biological samples. Here, we describe how to determine AEA and 2-AG levels in tiny samples of tissues and plasma through LC-MS/MS, by using very quick and easy-to-perform extraction procedures, with reduced solvent consumption.
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Endocannabinoides , Espectrometría de Masas en Tándem , Ácidos Araquidónicos , Cromatografía Liquida/métodos , Alcamidas Poliinsaturadas , Solventes , Espectrometría de Masas en Tándem/métodosRESUMEN
Recurrent Binge Eating (BE) episodes characterize several eating disorders. Here, we attempted to reassemble a condition closer to BE disorder, and we analyzed whether recurrent episodes might evoke molecular alterations in the hypothalamus of rats. The hypothalamus is a brain region which is sensitive to stress and relevant in motivated behaviors, such as food intake. A well-characterized animal model of BE, in which a history of intermittent food restriction and stress induce binge-like palatable food consumption, was used to analyze the transcriptional regulation of the endocannabinoid system (ECS). We detected, in rats showing the BE behavior, an up-regulated gene expression of cannabinoid type-1 receptor (CB1), sn-1-specific diacylglycerol lipase, as well as fatty acid amide hydrolase (Faah) and monoacylglycerol lipase. A selective reduction in DNA methylation was also observed at the promoter of Faah, which is consistent with the changes in the gene expression. Moreover, BE behavior in rats was associated with an increase in anandamide (AEA) levels. Our findings support the relevant role of the ECS in the regulation of food intake in rats subjected to repeated BE episodes, and, in particular, on AEA signaling, acting via CB1 and FAAH modulation. Notably, the epigenetic regulation of the Faah gene might suggest this enzyme as a possible target for developing new therapeutical approaches.
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Trastorno por Atracón , Ratas , Femenino , Animales , Trastorno por Atracón/genética , Epigénesis Genética , Endocannabinoides/metabolismo , Amidohidrolasas/genética , Amidohidrolasas/metabolismo , Monoacilglicerol Lipasas/genética , Monoacilglicerol Lipasas/metabolismo , Receptores de Cannabinoides/metabolismo , Hipotálamo/metabolismo , Receptor Cannabinoide CB1/genética , Receptor Cannabinoide CB1/metabolismo , Ingestión de AlimentosRESUMEN
Type 1 spinocerebellar ataxia (SCA1) is a progressive neurodegenerative disorder with no effective treatment to date. Using mice modeling SCA1, it has been demonstrated that a drug that amplifies mGlu1 receptor activation (mGlu1 receptor PAM, Ro0711401) improves motor coordination without the development of tolerance when cerebellar dysfunction manifests (i.e., in 30-week-old heterozygous ataxin-1 [154Q/2Q] transgenic mice). SCA1 is also associated with cognitive dysfunction, which may precede cerebellar motor signs. Here, we report that otherwise healthy, 8-week-old SCA1 mice showed a defect in spatial learning and memory associated with reduced protein levels of mGlu1α receptors, the GluN2B subunit of NMDA receptors, and cannabinoid CB1 receptors in the hippocampus. Systemic treatment with Ro0711401 (10 mg/kg, s.c.) partially corrected the learning deficit in the Morris water maze and restored memory retention in the SCA1 mice model. This treatment also enhanced hippocampal levels of the endocannabinoid, anandamide, without changing the levels of 2-arachidonylglycerol. These findings suggest that mGlu1 receptor PAMs may be beneficial in the treatment of motor and nonmotor signs associated with SCA1 and encourage further studies in animal models of SCA1 and other types of SCAs.