LC-IMS-HRMS for identification of biomarkers in untargeted metabolomics: The effects of pterostilbene and resveratrol consumption in liver steatosis, animal model.

Untargeted metabolomics with the combination of ion mobility separation coupled to high resolution mass spectrometry (IMS-HRMS) was applied to investigate the impact of resveratrol and pterostilbene supplementation on the metabolic fingerprint of the Wistar rats liver with induced liver steatosis. RP-LC and HILIC in both ionisation modes were employed to analyse the liver samples (n = 40) from Wistar rats fed with a high-fat and high-fructose diet, supplemented or not with resveratrol and pterostilbene. After univariate and multivariate statistical analysis, 34 metabolites were highlighted in the different diets and elucidated. Despite the structural similarity, different alterations in liver metabolism were observed by the supplementations. Resveratrol treatment was characterised by the alteration in metabolism of 17 lysophospholipids, while pterostilbene affected some vitamins and derivatives, among others. IMS has demonstrated great potential in the elucidation process thanks to the additional structural descriptor the CCS (Å2), providing more confidence in the identification.

CPEB alteration and aberrant transcriptome-polyadenylation lead to a treatable SLC19A3 deficiency in Huntington's disease.

Huntington's disease (HD) is a hereditary neurodegenerative disorder of the basal ganglia for which disease-modifying treatments are not yet available. Although gene-silencing therapies are currently being tested, further molecular mechanisms must be explored to identify druggable targets for HD. Cytoplasmic polyadenylation element binding proteins 1 to 4 (CPEB1 to CPEB4) are RNA binding proteins that repress or activate translation of CPE-containing transcripts by shortening or elongating their poly(A) tail. Here, we found increased CPEB1 and decreased CPEB4 protein in the striatum of patients and mouse models with HD. This correlated with a reprogramming of polyadenylation in 17.3% of the transcriptome, markedly affecting neurodegeneration-associated genes including PSEN1, MAPT, SNCA, LRRK2, PINK1, DJ1, SOD1, TARDBP, FUS, and HTT and suggesting a new molecular mechanism in neurodegenerative disease etiology. We found decreased protein content of top deadenylated transcripts, including striatal atrophy­linked genes not previously related to HD, such as KTN1 and the easily druggable SLC19A3 (the ThTr2 thiamine transporter). Mutations in SLC19A3 cause biotin-thiamine­responsive basal ganglia disease (BTBGD), a striatal disorder that can be treated with a combination of biotin and thiamine. Similar to patients with BTBGD, patients with HD demonstrated decreased thiamine in the cerebrospinal fluid. Furthermore, patients and mice with HD showed decreased striatal concentrations of thiamine pyrophosphate (TPP), the metabolically active form of thiamine. High-dose biotin and thiamine treatment prevented TPP deficiency in HD mice and attenuated the radiological, neuropathological, and motor HD-like phenotypes, revealing an easily implementable therapy that might benefit patients with HD.

Safety in the use of ultrasound in rheumatology during the COVID-19 Pandemic. Spanish Society of Rheumatology positioning paper. / Seguridad en el uso de la ecografía en reumatología durante la pandemia COVID-19. Documento de posicionamiento de la Sociedad Española de Reumatología.

Ultrasound is a widely implemented imaging modality in rheumatology practice that implies a great interaction between patient and professional. The COVID-19 pandemic requires a change in our clinical practice, through the adoption of new strategies that allow comprehensive care for our patients, guaranteeing the safety of both patients and healthcare professionals. OBJETIVE: Our objective was to develop practical recommendations, agreed among a panel of experts, on the use and safety of rheumatological ultrasound during the COVID-19 pandemic. METHODS: We performed a narrative review of the available literature. Based on the literature review, we produced preliminary recommendations that were subsequently agreed among a panel of experts using the Delphi methodology with a 1-5 Likert scale. Agreement for each recommendation was considered if 75% of the panel members scored the item ≥4 on the Likert scale. RESULTS: 5 overarching principles and 28 recommendations were issued and agreed among the panel. Group consensus was achieved in 100% of items. CONCLUSIONS: The document provides useful information about preventive measures in the practice of ultrasound in rheumatology in times of a COVID-19 pandemic based on the experience and literature available to date.

Direct and Fast Screening of New Psychoactive Substances Using Medical Swabs and Atmospheric Solids Analysis Probe Triple Quadrupole with Data-Dependent Acquisition.

New psychoactive substances (NPS) have become a serious public health problem, as they are continuously changing their structures and modifying their potency and effects on humans, and therefore, novel compounds are unceasingly appearing. One of the major challenges in forensic analysis, particularly related to the problem of NPS, is the development of fast screening methodologies that allow the detection of a wide variety of compounds in a single analysis. In this study, a novel application of the atmospheric solids analysis probe (ASAP) using medical swabs has been developed. The swab-ASAP was coupled to a triple quadrupole mass analyzer working under a data-dependent acquisition mode in order to perform a suspect screening of NPS in different types of samples as well as on surfaces. The compounds were automatically identified based on the observed fragmentation spectra using an in-house built MS/MS spectra library. The developed methodology was applied for the identification of psychoactive substances in research chemicals and herbal blends. The sensitivity of the method, as well as its applicability for surface analysis, was also assessed by identifying down to 1 µg of compound impregnated onto a laboratory table. Another remarkable application was the identification of cathinones and synthetic cannabinoids on the fingers of potential consumers. Interestingly, our data showed that NPS could be identified on the fingers after being in contact with the product and even after cleaning their hands by shaking off with a cloth. The methodology proposed in this paper can be applied for routine analyses of NPS in different matrix samples without the need to establish a list of target compounds prior to analysis.

Investigation on the consumption of synthetic cannabinoids among teenagers by the analysis of herbal blends and urine samples.

The use of synthetic cannabinoids (SCs), which escape conventional detection systems, may be a good alternative to elude routine drug analysis for cannabis. The detection of these drugs in urine is unusual due to their complete and fast metabolism, therefore requiring alternative strategies. In this work, an investigation has been made on SCs consumption by minors (less than 18 years old) in juvenile offenders' centres. 667 urine samples (from 127 minors) were collected after their permits with stay at home. We also studied the SCs from 7 herbal blends available at the smartshop frequented by the minors. Both, urine and herbal blends, were analysed by liquid chromatography coupled to high resolution mass spectrometry. The analysis of urine confirmed the absence of more than 200 SCs investigated. Thus, the focus was made on metabolites reported for those SCs identified in the herbal blends collected from the smart-shop. The major metabolites of XLR-11 and UR-144 (N-pentanoic acid and N-(5-hydroxypentyl)) were found in several urine samples. Apart from the main metabolites included in the initial searching, a thorough investigation of more metabolites for these SCs was additionally performed, including MS/MS experiments for the tentative identification of compounds detected in the urine samples. The 16 samples positive to the XLR-11 metabolites were assigned to 6 minors, only 2 of which had recognized consumption. On the basis of the results obtained, preventive and therapeutic interventions must be implemented to reduce the consumption of psychoactive substances and to improve the risk-perception of these substances by minors.

What about the herb? A new metabolomics approach for synthetic cannabinoid drug testing.

Synthetic cannabinoids (SCs) are consumed as legal alternative to cannabis and often allow passing drug-screening tests. Their rapid transience on the drug scene, combined with their mostly unknown metabolic profiles, creates a scenario with constantly moving analytical targets, making their monitoring and identification challenging. The development of fast screening strategies for SCs, not directly focused on their chemical structure, as an alternative to the commonly applied target acquisition methods, would be highly appreciated in forensic and public health laboratories. An innovative untargeted metabolomics approach, focused on herbal components commonly used for 'spice' products, was applied. Saliva samples of healthy volunteers were collected at pre-dose and after smoking herbal components and analysed by high-resolution mass spectrometry. The data obtained, combined with appropriate statistical analysis, allowed to highlight and elucidate two markers (scopoletin and N,N-bis(2-hydroxyethyl)dodecylamine), which ratio permitted to differentiate herbal smokers from non-smokers. The proposed strategy will allow discriminating potential positives, on the basis of the analysis of two markers identified in the herbal blends. This work is presented as a step forward in SC drug testing, promoting a smart first-line screening approach, which will allow reducing the number of samples to be further investigated by more sophisticated HRMS methods. Graphical abstract The development of an alternative, generic screening methods of synthetic cannabinoids, not directly based on the chemical structure, in order to provide fast response on its potential consumption.

Assessment of protected designation of origin for Colombian coffees based on HRMS-based metabolomics.

An untargeted metabolomics approach based on HRMS has been applied to Colombian green coffee to develop a discrimination model to highlight the most differential compounds. For this purpose, 41 green coffee samples of different genotypes collected from 5 regions were analysed. Samples were extracted with aqueous and organic solvents to cover a wide range of compounds. Sample extracts were randomly injected and data were pre-processed with XCMS software. PCA was used to verify quality control samples behaviour, and PLS-DA and DD-SIMCA were employed to create models for discrimination using VIP variable selection method. Thirteen different compounds correctly separate green coffee samples according to their origin, several related to the quality and health benefits of coffee. Model validation was achieved using both cross-validation and an additional set with coffee samples from different harvest year. The results reveal that UHPLC-(Q)ToF MS-based metabolomics is a suitable tool to develop food origin discrimination strategies.

Occurrence and fate of illicit drugs and pharmaceuticals in wastewater from two wastewater treatment plants in Costa Rica.

Chemical analysis of raw wastewater in order to assess the presence of biological markers entering a wastewater treatment plant can provide objective information about the health and lifestyle of the population connected to the sewer system. This work was performed in a tropical country of Central America, Costa Rica, with the aim of extending this knowledge to new world regions. This work is the first to report wastewater-based epidemiological data on the use of illicit drugs in this region of the world. Composite wastewater samples from the influents of two different wastewater treatment facilities and surface water samples from surrounding areas were collected applying the best practice protocol and analysed to investigate the occurrence and fate of selected illicit drugs of abuse and pharmaceuticals. Results showed the presence of chemical indicators of the classic drugs cocaine and cannabis at high concentration levels, besides the moderate presence of the opiates codeine and morphine. Neither the worldwide commonly used psychoactive substances of abuse such as synthetic phenethylamines, nor pharmaceuticals from the family of benzodiazepines were detected, demonstrating the spatial differences in drug use among different world regions. In addition, effluent wastewater samples were analysed and compared to influent concentrations in order to evaluate the decrease in concentration of the targeted analytes through two treatment technologies. As a final step, a wide-scope qualitative screening, including hundreds of suspect compounds, was applied in order to have a better knowledge on the presence of pharmaceuticals in waters and to assess the potential impact of the treated wastewater into the receiving aquatic ecosystems.

Comprehensive monitoring of organic micro-pollutants in surface and groundwater in the surrounding of a solid-waste treatment plant of Castellón, Spain.

The solid-waste treatment plant of RECIPLASA is located in the municipality of Onda (Castellón province), which is an important agricultural area of Spain, with predominance of citrus crops. In this plant, all urban solid wastes from the town of Castellón (around 200,000 inhabitants) and other smaller towns as Almassora, Benicàssim, Betxí, Borriana, L'Alcora, Onda and Vila-Real are treated. In order to evaluate the potential impact of this plant on the surrounding water, both surface and groundwater, a comprehensive monitoring of organic pollutants has been carried out along 2011, 2012 and 2013. To this aim, an advanced analytical strategy was applied for wide-scope screening, consisting on the complementary use of liquid chromatography (LC) and gas chromatography (GC) coupled to mass spectrometry (MS) with quadrupole (Q)-time of flight analyser (TOF). A generic solid-phase extraction with Oasis HLB cartridges was applied prior to the chromatographic analysis. The screening included more than 1500 organic pollutants as target compounds, such as pesticides, pharmaceuticals, veterinary drugs, drugs of abuse, UV-filters, polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), polybrominated diphenyl ethers (PBDEs), among others. Pesticides, mainly herbicides, were the compounds more frequently detected. Other compounds as antioxidants, cosmetics, drugs of abuse, PAHs, pharmaceuticals and UV filters, were also identified in the screening though at much lower frequency. Once the screening was made, quantitative analysis focused on the compounds more frequently detected was subsequently applied using LC coupled to tandem MS with triple quadrupole analyser. In this way, up to 24 pesticides and transformation products (TPs), 7 pharmaceuticals, one drug of abuse and its metabolite could be quantified at sub-ppb concentrations. Along the three years of study, ten compounds were found at concentrations higher than 0.1µg/L. Most of them were pesticides and TPs, a fact that illustrates that the main source of pollution seems to be the agricultural activities in this area.

Quadrupole-time-of-flight mass spectrometry screening for synthetic cannabinoids in herbal blends.

'Legal highs' are novel substances which are intended to elicit a psychoactive response. They are sold from 'head shops', the internet and from street suppliers and may be possessed without legal restriction. Several months ago, a 19-year-old woman came searching for medical treatment as she had health problems caused by smoking legal highs. The substances were sold as herbal blends in plastic bags under four different labels. In this work, samples of these herbal blends have been analysed to investigate the presence of psychoactive substances without any reference standard being available at the laboratory. A screening strategy for a large number of synthetic and natural cannabinoids has been applied based on the use of ultra-high pressure liquid chromatography coupled to quadrupole-time of flight mass spectrometry (UHPLC-QTOF MS) under MS(E) mode. A customized home-made database containing literature-based exact masses for parent and product ions of around 200 synthetic and natural cannabinoids was compiled. The presence of the (de)protonated molecule measured at its accurate mass was evaluated in the samples. When a peak was detected, collision-induced dissociation fragments and characteristic isotopic ions were also evaluated and used for tentative identification. After this tentative identification, four synthetic cannabinoids (JWH-081, JWH-250, JWH-203 and JWH-019) were unequivocally confirmed by subsequent acquisition of reference standards. The presence in the herbal blends of these synthetic cannabinoids might explain the psychotic and catatonic symptoms observed in the patient, as JWH compounds could act as potent agonists of CB1 and CB2 receptors located in the Limbic System and Basal ganglia of the human brain.

Aplicación del OLEOZÓN en el tratamiento de las pericoronaritis / Usage of Oleozón in the treatment of pericoronaritis

La atención primaria se orienta hacia los principales problemas de salud de la comunidad, entre ellos los de urgencias estomatológicas, donde es atendida la pericoronaritis, siendo el tratamiento indicado la aplicación de sustancias cáusticas (ácido tricloroacético), que pueden ocasionar daños en los tejidos dentarios. Teniendo en cuenta las características de la pericoronaritis, se utilizó la ozonoterapia, realizando un ensayo clínico fase III, aleatorizado, controlado y abierto, lo cual permitió estudiar la evolución microbiológica después de aplicado el OLEOZÓN en el tratamiento de la pericoronaritis, en la Clínica Estomatológica Docente César Escalante, del municipio Matanzas, de enero 2003 a enero 2008. El universo de estudio fueron los pacientes con pericoronaritis, y la muestra abarcó 90 pacientes, conformándose dos grupos, uno experimental, tratado con Oleozón, y otro control, donde se empleó el tratamiento convencional. La eficacia del OLEOZÓN resultó ser alta en los pacientes tratados(AU)

Primary care is directed to the main health problems of the community, and among them stomatologic emergencies where pericoronaritis is attended, being the indicated treatment the application of caustic substances (trichloroacetic acid), that may damage dental tissues. Considering the characteristics of pericoronaritis, we used ozone therapy, carrying out a Phase III randomized, controlled and open clinical trial, that allowed studying the microbiologic evolution after applying Oleozón in the treatment of pericoronaritis, at the Teaching Stomatologic Clinic Cesar Escalante, municipality of Matanzas, from January 2003 to January 2008. The universe of study was the patients with pericoronaritis, and the sample covered 90 patients. Two groups were formed, one experimental, treated with Oleozon, and the other control, where we used the conventional treatment. The OLEOZÓN efficacy was high in the treated patients(AU)

Gas chromatography-mass spectrometric determination of polybrominated diphenyl ethers in complex fatty matrices from aquaculture activities.

Gas chromatography coupled to mass spectrometry in negative chemical ionization mode (GC-(NCI)MS) has been applied to the quantification and reliable identification of polybrominated diphenyl ethers (PBDEs) in animal and vegetable samples from aquaculture activities. Matrices analyzed included fish fillet, fish feed, fish oil and linseed oil, their fat content ranged from 5% to 100%. Solid-phase extraction (SPE) (using Florisil and silica cartridges) and normal-phase high performance liquid chromatography were tested for an efficient clean-up in order to obtain sample extracts free of interfering compounds. Combining sulphuric acid digestion and SPE with Florisil led to the highest efficiency in the elimination of interferences from the extracts. The sample procedure developed, together with the application of GC-(NCI)MS for measurement, led to the satisfactory determination of PBDEs at microg kg(-1) levels in complex aquaculture matrices with high lipid content. The use of a short and thin film-thickness fused-silica capillary column allowed to determine the problematic BDE 209 with satisfactory results. Three m/z ions were acquired for each analyte, which ensured a reliable identification of compounds detected in samples.

Evolución clínica de las pericoronaritis tratadas con OLEOZON® en urgencias estomatológicas / Clinical evolution of pericoronaritis treated with OLEOZÓN® in stomatologic urgencias

La pericoronaritis es una patología bucal que aparece con frecuencia en consultas de urgencias estomatológicas. Se considera un proceso infeccioso agudo caracterizado por inflamación del tejido blando que rodea el diente retenido, lo cual provoca efectos indeseables en el paciente. El tratamiento indicado es la aplicación de sustancias caústicas que pueden ocasionar daños en los tejidos dentarios, por lo cual se decidió utilizar la ozonoterapia para realizar un ensayo clínico fase III, aleatorizado, controlado y abierto, lo cual permitió estudiar la eficacia del OLEOZON® en el tratamiento de la Pericoronaritis, en la Clínica Estomatológica Docente César Escalante, del municipio Matanzas, de enero 2003 a enero de 2008. El universo de estudio fueron los pacientes que en ese período asistieron con Pericoronaritis a la consulta de urgencia, y la muestra abarcó 90 pacientes, conformándose dos grupos: uno experimental, donde se aplicó el OLEOZON®, y otro control, donde se empleó el tratamiento convencional. Se caracterizaron los grupos desde el punto de vista sociodemográfico y clínico, determinándose la eficacia del OLEOZON®, la cual resultó ser alta en los pacientes tratados...(AU)

Pericoronaritis is an oral pathology frequently found in stomatologic urgency consultations. It is considered an acute infectious process characterized by inflammation of the soft tissues surrounding the retained tooth, causing unwanted effects in the patient. The indicated treatment is applying caustic substances that may damage dental tissues, so we decided using ozonotherapy to develop a clinical assay Phase III, randomized, controlled and open, allowing us to study the efficacy of OLEOZÓN® in treating Pericoronaritis at the Teaching Stomatologic Clinic Cesar Escalante, municipality of Matanzas, from january 2003 to january 2008. The universe of study were the patients assisting to the urgency consultation with Pericoronaritis in that period of time, and the sample were 90 patients, divided in two groups, an Experimental one where OLEOZÓN® was applied, and a Control one where the conventional treatment was used. Both groups were characterized from the sociodemographic and clinic points of view, stating the efficacy of OLEOZÓN®, high in treated patients...(AU)

Evolución clínica de las pericoronaritis tratadas con OLEOZON® en urgencias estomatológicas / Clinical evolution of pericoronaritis treated with OLEOZÓN® in stomatologic urgencias

La pericoronaritis es una patología bucal que aparece con frecuencia en consultas de urgencias estomatológicas. Se considera un proceso infeccioso agudo caracterizado por inflamación del tejido blando que rodea el diente retenido, lo cual provoca efectos indeseables en el paciente. El tratamiento indicado es la aplicación de sustancias caústicas que pueden ocasionar daños en los tejidos dentarios, por lo cual se decidió utilizar la ozonoterapia para realizar un ensayo clínico fase III, aleatorizado, controlado y abierto, lo cual permitió estudiar la eficacia del OLEOZON® en el tratamiento de la Pericoronaritis, en la Clínica Estomatológica Docente César Escalante, del municipio Matanzas, de enero 2003 a enero de 2008. El universo de estudio fueron los pacientes que en ese período asistieron con Pericoronaritis a la consulta de urgencia, y la muestra abarcó 90 pacientes, conformándose dos grupos: uno experimental, donde se aplicó el OLEOZON®, y otro control, donde se empleó el tratamiento convencional. Se caracterizaron los grupos desde el punto de vista sociodemográfico y clínico, determinándose la eficacia del OLEOZON®, la cual resultó ser alta en los pacientes tratados.

Pericoronaritis is an oral pathology frequently found in stomatologic urgency consultations. It is considered an acute infectious process characterized by inflammation of the soft tissues surrounding the retained tooth, causing unwanted effects in the patient. The indicated treatment is applying caustic substances that may damage dental tissues, so we decided using ozonotherapy to develop a clinical assay Phase III, randomized, controlled and open, allowing us to study the efficacy of OLEOZÓN® in treating Pericoronaritis at the Teaching Stomatologic Clinic Cesar Escalante, municipality of Matanzas, from january 2003 to january 2008. The universe of study were the patients assisting to the urgency consultation with Pericoronaritis in that period of time, and the sample were 90 patients, divided in two groups, an Experimental one where OLEOZÓN® was applied, and a Control one where the conventional treatment was used. Both groups were characterized from the sociodemographic and clinic points of view, stating the efficacy of OLEOZÓN®, high in treated patients.

Use of ultra-high-pressure liquid chromatography-quadrupole time-of-flight MS to discover the presence of pesticide metabolites in food samples.

In this paper we illustrate the use of two different methodologies to investigate the presence of pesticide metabolites in parent pesticide-positive food samples, using ultra-high-pressure liquid chromatography coupled to hybrid quadrupole time-of-flight (UHPLC-QTOF) mass spectrometry. First, a common fragmentation pathway between the parent pesticide and its metabolites has been considered to search for metabolites in two positive market samples (imazalil in lemon, chlorpyrifos in grape). Secondly, olive oil samples from field residue trials were used for automated application of comparative software (MetaboLynx), which was used with treated and untreated samples to search for expected and unexpected metabolites of phosmet. One of the main objectives when using these approaches was to avoid the tedious manual searching for potential metabolites within the huge amount of information contained in the total ion chromatogram acquired by TOF MS. The common fragmentation approach applied to TOF MS full-acquisition data, considering an enhanced fragmentation in the collision cell, has allowed the discovery of two metabolites of imazalil (1-[2-(2,4-dichlorophenyl)-2-hydroxyethyl]-1H-imidazole and 1-[2-(2,4-dichlorophenyl)-2-oxoethyl]-1H-imidazole) in a lemon positive sample, as well as another two metabolites of chlorpyrifos (chlorpyrifos-oxon and 3,5,6-trichloro-2-pyridinol) in a grape positive sample. Moreover, MetaboLynx application to TOF MS data, without promoting fragmentation, from treated and untreated olive oil samples has been helpful in detecting the metabolite phosmet-oxon. In both strategies, every metabolite detected by TOF MS was confirmed using QTOF and/or triple quadrupole instruments. Accurate masses given by TOF MS together with the valuable information on product ions given by QTOF MS/MS experiments were crucial for the unambiguous identification of metabolites.