Determination of synthetic opioids in oral fluid samples using fabric phase sorptive extraction and gas chromatography-mass spectrometry

New psychoactive substances (NPS) continue to emerge in the drug market every year, becoming a global threat to public health and safety. These compounds are mostly synthetic cannabinoids and designer cathinones. However, synthetic opioids have appeared on the recreational drug markets in recent years, particularly fentanyl and its derivatives (“fentanyls”). Fentanyl and its analogs are related to harmful intoxications and an increase in opioid-related mortality in many countries, such as in the United States and Europe in the last years. Taking the drug related global crisis into consideration, this work developed and validated an effective and sensitive method based on fabric phase sorptive extraction (FPSE) followed by gas chromatography-mass spectrometry (GC-MS) for the simultaneous determination of 11 fentanyl analogs in oral fluid samples. The extraction was carried out using a sol-gel Carbowax 20 M sorbent immobilized on 100% cellulose fabric substrate and using ethyl acetate as the desorption solvent. The limits of detection (LODs) and quantification (LOQs) ranged from 1 to 15 ng mL−1 and 5 to 50 ng mL−1, respectively. Intra-day and inter-day precision were found within 8.2% and 8.6%, respectively, while accuracy ranged from -5.5 to 9.1%, in accordance with the established criteria. The absolute recovery values were in the range of 94.5%–109.1%. The validated method demonstrated its great potential to detect and quantify fentanyl analogs in possible forensic work and off-site analysis in road traffic casesThis work was supported by the Spanish Ministry of Science, Innovation, and Universities (Project RTI2018‐096450‐B‐I00) and FEDER fundsS

Comprehensive metabolite profiling of Solanum tuberosum L.(potato) leaves by HPLC-ESI-QTOF-MS

The objective of this work was to study the non-targeted metabolite profiling of potato leaves using high performance liquid chromatography coupled to quadrupole-time of flight mass spectrometry (HPLC-ESI-QTOF-MS). The mass accuracy, true isotopic pattern in both MS and MS/MS spectra provided by QTOF-MS made possible the tentative identification of 109 compounds present in potato leaves, including organic acids, amino acids and derivatives, phenolic acids, flavonoids, iridoids, oxylipins and other polar and semi-polar compounds. Among them, 32 compounds have been found for the first time in potato leaf and in the Solanaceae family. Quinic acid and its derivatives represented more than 45% of the bioactive compounds quantified in the extract. Derivatives of hydroxybenzoic acid and gentisic acid were also founded at considerable concentrations. This study shed light on the composition of potato leaf extract and will serve as a base for further research into activities of the various compounds found in this matrix which has demonstrated a potential use as functional ingredients

From Olive Fruits to Olive Oil: Phenolic Compound Transfer in Six Different Olive Cultivars Grown under the Same Agronomical Conditions

Phenolic compounds are responsible of the nutritional and sensory quality of extra-virgin olive oil (EVOO). The composition of phenolic compounds in EVOO is related to the initial content of phenolic compounds in the olive-fruit tissues and the activity of enzymes acting on these compounds during the industrial process to produce the oil. In this work, the phenolic composition was studied in six major cultivars grown in the same orchard under the same agronomical and environmental conditions in an effort to test the effects of cultivars on phenolic composition in fruits and oils as well as on transfer between matrices. The phenolic fractions were identified and quantified using high-performance liquid chromatography-diode array detector-time-of-flight-mass spectrometry. A total of 33 phenolic compounds were determined in the fruit samples and a total of 20 compounds in their corresponding oils. Qualitative and quantitative differences in phenolic composition were found among cultivars in both matrices, as well as regarding the transfer rate of phenolic compounds from fruits to oil. The results also varied according to the different phenolic groups evaluated, with secoiridoids registering the highest transfer rates from fruits to oils. Moreover, wide-ranging differences have been noticed between cultivars for the transfer rates of secoiridoids (4.36%–65.63% of total transfer rate) and for flavonoids (0.18%–0.67% of total transfer rate). ‘Picual’ was the cultivar that transferred secoiridoids to oil at the highest rate, whereas ‘Changlot Real’ was the cultivar that transferred flavonoids at the highest rates instead. Principal-component analysis confirmed a strong genetic effect on the basis of the phenolic profile both in the olive fruits and in the oils.This research was partly funded by project P11-AGR-7301 of the Andalusian Regional Government Council of Innovation and Science and the Spanish Ministry of Economy and Competitiveness. The analytical part was financially supported by Spanish Ministry of Education and Science (AGL2015-67995-C3) and the Andalusian Regional Government Council of Innovation and Science (P10-FQM-6563, P11-CTS-7625). The authors thank the International Olive Oil Council (IOC) for the grant (Ref.: T1 6-Doct 1/12) and the Ministry of Education and Competitiveness for the postdoctoral contract Juan de la Cierva (JCI-2012-12566)

Perimeter and carvacrol-loading regulate angiogenesis and biofilm growth in 3D printed PLA scaffolds

rvacrol is a natural low-cost compound derived from oregano which presents anti-bacterial properties against both Gram-positive and Gram-negative bacteria. In this work, carvacrol-loaded PLA scaffolds were fabricated by 3D printing as platforms to support bone tissue regeneration while preventing biofilm development. Scaffolds were printed with or without a perimeter (lateral wall) mimicking the cortical structure of bone tissue to further evaluate if the lateral interconnectivity could affect the biological or antimicrobial properties of the scaffolds. Carvacrol incorporation was performed by loading either the PLA filament prior to 3D printing or the already printed PLA scaffold. The loading method determined carvacrol localization in the scaffolds and its release profile. Biphasic profiles were recorded in all cases, but scaffolds loaded post-printed released carvacrol much faster, with 50–80% released in the first day, compared to those containing carvacrol in PLA filament before printing which sustained the release for several weeks. The presence or absence of the perimeter did not affect the release rate, but total amount released. Tissue integration and vascularization of carvacrol-loaded scaffolds were evaluated in a chorioallantoic membrane model (CAM) using a novel quantitative micro-computed tomography (micro-CT) analysis approach. The obtained results confirmed the CAM tissue ingrowth and new vessel formation within the porous structure of the scaffolds after 7 days of incubation, without leading to hemorrhagic or cytotoxic effects. The absence of lateral wall facilitated lateral integration of the scaffolds in the host tissue, although increased the anisotropy of the mechanical properties. Scaffolds loaded with carvacrol post-printing showed antibiofilm activity against Staphylococcus aureus and Pseudomonas aeruginosa as observed in a decrease in CFU counting after biofilm detachment, changes in metabolic heat measured by calorimetry, and increased contact killing efficiency. In summary, this work demonstrated the feasibility of tuning carvacrol release rate and the amount released from PLA scaffolds to achieve antibiofilm protection without altering angiogenesis, which was mostly dependent on the perimeter density of the scaffoldsThis work was supported by MCIN/AEI/http://dx.doi.org/10.13039/501100011033 [PID2020-113881RB-I00], FEDER, and Xunta de Galicia [ED431C 2020/17, ED481D-2021-014]. Xián Farto-Vaamonde acknowledges Xunta de Galicia for a predoctoral research fellowship [ED481A-2018/073]S

Seasonal and interannual changes in Ca and Mg of dripping waters in Kaite Cave (Spain)

Depto. de Geodinámica, Estratigrafía y PaleontologíaFac. de Ciencias GeológicasTRUEpu

Present-day calcite deposition in two caves of N Spain (Kaite and Cueva Mayor): Factors affecting calcite growth and fabric

Depto. de Geodinámica, Estratigrafía y PaleontologíaFac. de Ciencias GeológicasTRUEpu

V232D Mutation in Patients With Cystic Fibrosis: Not So Rare, Not So Mild

The frequency of some Cystic Fibrosis (CF) Transmembrane Conductance Regulator gene (CFTR) mutations varies between populations. Genetic testing during newborn screening (NBS) for CF can identify less common mutations with low clinical expression in childhood and previously considered mild but not fully characterized, such as the mutation p.Val232Asp (c.695T > A). The aim of this study was to describe CF patients with the V232D mutation. We identify CF children with the V232D mutation detected by NBS and compare them with CF adults with this mutation whose diagnosis was prompted by clinical symptoms in the same period. We studied clinical, biochemical, spirometric, and prognostic features in both populations. NBS program tested 276,523 children during a period of 14 years (2003-2017) and identified 54 cases of CF. Six children (11%) had the V232D mutation. Over the same period, 5 adults (age 37.6 ± 16.29 years old) with symptoms of CF and this mutation were also diagnosed. Follow-up duration was mean 10.1 years for adults and mean 6.5 years for children. In the adult group, lung function was impaired at diagnosis in all patients (Forced Expiratory Volume1-FEV1-67.12% ± 13.09) and worsened in children tested during evolution (FEV1first: 113%; FEV1last: 64%). Pancreatic insufficiency was present in adult group, with recurrent pancreatitis in 1 present. Although with less clinical expression in children, V232D is associated with pulmonary and pancreatic involvement during adulthood and CF cannot be considered mild. This mutation is present in 11% of all patients diagnosed with CF in our region. Its inclusion in some NBS programs should be taken into account in order to improve the prognosis of affected children.S

Terpene down-regulation triggers defense responses in transgenic oranges leading to resistance against fungal pathogens

Terpenoid volatiles are isoprene compounds that are emitted by plants to communicate with the environment. In addition to their function in repelling herbivores and attracting carnivorous predators in green tissues, the presumed primary function of terpenoid volatiles released from mature fruits is the attraction of seed-dispersing animals. Mature oranges (Citrus sinensis) primarily accumulate terpenes in peel oil glands, with D-limonene accounting for approximately 97% of the total volatile terpenes. In a previous report, we showed that down-regulation of a D-limonene synthase gene alters monoterpene levels in orange antisense (AS) fruits, leading to resistance against Penicillium digitatum infection. A global gene expression analysis of AS versus empty vector (EV) transgenic fruits revealed that the down-regulation of D-limonene up-regulated genes involved in the innate immune response. Basal levels of jasmonic acid were substantially higher in the EV compared with AS oranges. Upon fungal challenge, salicylic acid levels were triggered in EV samples, while jasmonic acid metabolism and signaling were drastically increased in AS orange peels. In nature, D-limonene levels increase in orange fruit once the seeds are fully viable. The inverse correlation between the increase in D-limonene content and the decrease in the defense response suggests that D-limonene promotes infection by microorganisms that are likely involved in facilitating access to the pulp for seed-dispersing frugivores.This work was supported by the Fondo Europeo de Desarrollo Regional and the Ministry of Science and Innovation of Spain (grant no. AGL2009-08052), by the Fundo de Defesa da Citricultura, and by the Instituto Valenciano de Investigaciones Agrarias, Spain (Ph.D. fellowship to A.R.).Rodríguez, A.; Shimada, T.; Cervera, M.; Alquézar, B.; Gadea Vacas, J.; Gómez-Cadenas, A.; De Ollas, CJ.... (2014). Terpene down-regulation triggers defense responses in transgenic oranges leading to resistance against fungal pathogens. Plant Physiology. 164(1):321-339. https://doi.org/10.1104/pp.113.224279S321339164

Indicadores de observación y competencias en el Prácticum Grados Infantil y Primaria: centro y aula

El trabajo que se presenta está enmarcado en la RED Diseño y desarrollo de una Guía observacional para la implementación del Prácticum de los Grados de Infantil y Primaria, dentro del Programa de REDES de investigación en docencia universitaria del ICE/ Universidad de Alicante. Tomando como referencia las competencias de la asignatura Prácticum que se desarrolla a lo largo de varios periodos, se han establecido una serie de indicadores de observación a nivel de centro y de aula, que nos permitan configurar una Guía observacional como instrumento de observación y análisis de la realidad escolar durante el desarrollo del Prácticum. La metodología utilizada se ha centrado en el trabajo colaborativo de los integrantes de la RED, desde sus diversas experiencias y aportaciones en sus campos profesionales. Los primeros resultados han determinado los seis grandes bloques de indicadores de esta Guía: la organización y gestión del centro; la organización y gestión del aula; el clima del aula y el alumnado; el planteamiento metodológico y la evaluación; la atención a las n.e.e.; y la participación de las familias

Un estudio en RED: Guía observacional en el Prácticum Grados Infantil y Primaria

El trabajo que se presenta está enmarcado en la RED Diseño y desarrollo de una Guía observacional para la implementación del Prácticum de los Grados de Infantil y Primaria, dentro del Programa de REDES de investigación en docencia universitaria del ICE/ Universidad de Alicante. Tomando como referencia las competencias de la asignatura Prácticum que se desarrolla a lo largo de varios periodos, se han establecido una serie de indicadores de observación a nivel de centro y de aula, que nos permitan configurar una Guía observacional como instrumento de observación y análisis de la realidad escolar durante el desarrollo del Prácticum. La metodología utilizada se ha centrado en el trabajo colaborativo de los integrantes de la RED, desde sus diversas experiencias y aportaciones en sus campos profesionales. Los primeros resultados han determinado los seis grandes bloques de indicadores de esta Guía: la organización y gestión del centro; la organización y gestión del aula; el clima del aula y el alumnado; el planteamiento metodológico y la evaluación; la atención a las n.e.e.; y la participación de las familias