Validation of a method for cylindrospermopsin determination in vegetables: Application to real samples such as lettuce (lactuca sativa l.)

Reports on the occurrence of the cyanobacterial toxin cylindrospermopsin (CYN) have increased worldwide because of CYN toxic effects in humans and animals. If contaminated waters are used for plant irrigation, these could represent a possible CYN exposure route for humans. For the first time, a method employing solid phase extraction and quantification by ultra-performance liquid chromatography–tandem mass spectrometry (UPLC-MS/MS) of CYN was optimized in vegetables matrices such as lettuce (Lactuca sativa). The validated method showed a linear range, from 5 to 500 ng CYN g−1 of fresh weight (f.w.), and detection and quantitation limits (LOD and LOQ) of 0.22 and 0.42 ng CYN g−1 f.w., respectively. The mean recoveries ranged between 85 and 104%, and the intermediate precision from 12.7 to 14.7%. The method showed to be robust for the three different variables tested. Moreover, it was successfully applied to quantify CYN in edible lettuce leaves exposed to CYN-contaminated water (10 µg L−1), showing that the tolerable daily intake (TDI) in the case of CYN could be exceeded in elderly high consumers. The validated method showed good results in terms of sensitivity, precision, accuracy, and robustness for CYN determination in leaf vegetables such as lettuce. More studies are needed in order to prevent the risks associated with the consumption of CYN-contaminated vegetables. © 2018 by the authors. Licensee MDPI, Basel, Switzerland.Acknowledgments: The authors would like to acknowledge the Ministerio de Economía y Competitividad of Spain (AGL2015-64558-R, MINECO/FEDER, UE) for its financial support and the FCT Project UID/Multi/04423/2013. The authors also gratefully acknowledge the Spanish Ministerio de Economia y Competitividad for the grant FPI (BES-2016-078773) awarded to Leticia Díez-Quijada Jiménez. A. Campos work was supported by a postdoctoral grant (SFRH/BPD/103683/2014) from FCT

Does Size Matters? Relationships among Social Dominance and Some Morphometric Traits upon Out-of-Season Reproductive Outcomes in Anestrus Dairy Goats Treated with P4 + eCG

The possible role of the social rank (R) (i.e., low-LSR, middle-MSR, or high-HSR) in anestrus goats exposed to a P4 + eCG (D) (i.e., 100 or 350 IU) estrus induction protocol (EIP) was evaluated. Goats (Alpine-Saanen-Nubian × Criollo; n = 70; 25° North) managed under stall-fed conditions were all ultrasound evaluated to confirm anestrous status, while the social rank was determined 30 d prior to the EIP. The response variables included estrus induction (EI, %), duration of estrus (DUR, h), ovulation rate (OR, n), live weight (LW, kg), thoracic perimeter (TP, cm), thoracic diameter (TD, cm), body length (BL, cm), height at withers (HW, cm), beard length (BEA, cm), compactness index (COM, cm), and anamorphosis index (ANA, cm), as affected by R, D, and the R × D interaction were evaluated, while the correlation coefficients among reproductive and morphometric variables were quantified. An R × D interaction (p 0.05) with any response variable, either reproductive or morphometric. As expected, LW had high correlation coefficients (p < 0.01) with TP (0.86), TD (0.88), BL (0.82), HW (0.75), BEA (0.51), COM (0.97), and ANA (0.75). In general, the morphometric variables as a whole demonstrated important correlation coefficients among them (p < 0.01), ranging from 0.38 up to 0.84. To estimate the importance of the morphometric differences between social rank upon estrus induction, a principal component (PC) analysis was performed based on the correlation matrix derived from the corporal measurements. The PC1 and PC2 explained 70.3% and 17.6% of the morphometric variation, respectively. The PC1 was a measure of the goat size (i.e., small, medium, large) and its association with estrus occurrence was evaluated using a logistic regression model; the bigger the goat, the increased probability of being in estrus, by up to five times compared to small goats. Our results confirm that the higher social ranked, larger goats amalgamated some fundamental factors to be successful: aggressiveness, primacy to food access, augmented live weight, and corporal size; all of these were able to modulate out-of-season reproductive success in crossbred dairy goats subjected to an estrus induction protocol and managed under stall-fed conditions in Northern Mexico

El uso de una escape room como recurso docente en la Facultad de Farmacia

[EN] Escape Rooms are educational resources that are being used by many educators, fostering elements of collaboration that help in social skills development. In this study, a multidisciplinary Escape Room has been developed as an educational tool in order to improve students learning and prepare them for the future of professional practice. 145 students and 12 professors belonging to 8 different areas of knowledge from the Faculty of Pharmacy of the University of Seville participated in this activity. Through this innovation, a student-teacher feedback has been created, improving their relationship and learning from each other. The results obtained through satisfaction surveys show the great acceptance that the escape room had among the students, being the study valued with an overall score of 4.83/5. Although 93% of the students considered the activity of high/very high difficulty, 100% of them emphasize that this tool encourages teamwork and 86.9% consider that it reinforces knowledge. This study has awakened students motivation and interest in the subjects involved, strengthening learning and knowledge of the fundamental concepts acquired, and promoting teamwork and problem solving.[ES] Las Escape Rooms son un tipo de recurso que está siendo utilizado por muchos educadores, propiciando elementos de colaboración que ayudan a desarrollar habilidades sociales. En este estudio se ha desarrollado una Escape Room multidisciplinar como herramienta educativa con el fin de mejorar el aprendizaje de los estudiantes y prepararlos para el futuro de la práctica profesional. Participaron 145 alumnos y 12 profesores adscritos a 8 áreas de conocimiento distintas de la Facultad de Farmacia de la Universidad de Sevilla. Mediante esta innovación se ha producido una retroalimentación alumno-profesor, mejorando su relación y aprendiendo mutuamente. Los resultados obtenidos mediante encuestas de satisfacción muestran la gran aceptación que ha tenido la Escape Room entre los alumnos, siendo el estudio valorado con una puntuación general de 4,83/5. A pesar de que el 93% de los alumnos consideró la actividad de alta/muy alta dificultad, el 100% de los mismos destacan que esta herramienta fomenta el trabajo en equipo y el 86,9% considera que refuerza los conocimientos. Este estudio ha despertado en los alumnos la motivación e interés por las asignaturas implicadas, fortaleciendo el aprendizaje y conocimiento de los conceptos fundamentales adquiridos, y propiciando el trabajo en equipo y la resolución de problemas.Gutiérrez Praena, D.; Rios -Reina, R.; Ruiz, R.; Talero, E.; Callejón, R.; Callejón, RM.; Casas, M.... (2019). El uso de una escape room como recurso docente en la Facultad de Farmacia. En IN-RED 2019. V Congreso de Innovación Educativa y Docencia en Red. Editorial Universitat Politècnica de València. 1145-1155. https://doi.org/10.4995/INRED2019.2019.10356OCS1145115

Phase I clinical trial in healthy adults of a nasal vaccine candidate containing recombinant hepatitis B surface and core antigens

SummaryBackgroundThe nasal vaccine candidate (NASVAC), comprising hepatitis B virus (HBV) surface (HBsAg) and core antigens (HBcAg), has been shown to be highly immunogenic in animal models.MethodsA phase I double-blinded, placebo-controlled randomized clinical trial was carried out in 19 healthy male adults with no serologic markers of immunity/infection to HBV. This study was aimed at exploring the safety and immunogenic profile of nasal co-administration of both HBV recombinant antigens. The trial was performed according to Good Clinical Practice guidelines. Participants ranged in age from 18 to 45 years and were randomly allocated to receive a mixture of 50μg HBsAg and 50μg HBcAg or 0.9% physiologic saline solution, as a placebo, via nasal spray in a five-dose schedule at 0, 7, 15, 30, and 60 days. A total volume of 0.5ml was administered in two dosages of 125μl per nostril. Adverse events were actively recorded 1h, 6h, 12h, 24h, 48h, 72h, 7 days and 30 days after each dose. Anti-HBs and anti-HBc titers were evaluated using corresponding ELISA kits at days 30 and 90.ResultsThe vaccine candidate was safe and well tolerated. Adverse reactions included sneezing (34.1%), rhinorrhea (12.2%), nasal stuffiness (9.8%), palate itching (9.8%), headache (9.8%), and general malaise (7.3%). These reactions were all self-limiting and mild in intensity. No severe or unexpected events were recorded during the trial. The vaccine elicited anti-HBc seroconversion in 100% of subjects as early as day 30 of the immunization schedule, while a seroprotective anti-HBs titer (≥10IU/l) was at a maximum at day 90 (75%). All subjects in the placebo group remained seronegative during the trial.ConclusionThe HBsAg–HBcAg vaccine candidate was safe, well tolerated and immunogenic in this phase I study in healthy adults. To our knowledge, this is the first demonstration of safety and immunogenicity for a nasal vaccine candidate comprising HBV antigens

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

Pirólisis analítica (Py-GC/MS) para la determinación de cilindrospermopsina en músculo de pescado cocinado

Póster presentado en el XXIII Congreso Español de Toxicología y VII Iberoamericano. 26- 28 Junio 2019La cianotoxina cilindrospermopsina (CYN) es una sustancia hepatotóxica cada vez más frecuente a nivel mundial que puede acumularse en una gran variedad de animales acuáticos y transmitirse fácilmente lo largo de la cadena alimentaria. Por ello, es importante disponer de métodos analíticos adecuados que permitan su detección en muestras de pescado contaminado, tanto crudo como ya cocinado. El objetivo de este trabajo es optimizar un método basado en pirólisis analítica para su aplicación en la detección de CYN y sus productos de descomposición en carne de pescado cruda y cocinada. Para ello, utilizamos músculo de tilapia (Oreochromis niloticus) en filetes (4 g, n=5) a los que se les inyectó CYN pura (50 ng CYN/g peso seco) y se cocinaron durante 2 min mediante diferentes técnicas como el asado, microondas, hervido o vapor. Se mantuvo un grupo sin cocinar como control positivo y otro sin CYN y sin cocinar como control negativo. Todas las muestras se congelaron (-80°C) y se liofilizaron hasta ser analizadas. La Py-GC/MS se llevó a cabo en un pirolizador de doble disparo acoplado a un sistema de cromatografía de gases. Las muestras de pescado liofilizado (1,8-2,9 mg) se situaron en un crisol de pirólisis y se sometieron a 350 °C durante 1 min en ausencia de oxígeno. En las condiciones cromatográficas utilizadas la Py-GC/MS directa permitió la detección de la CYN (PM 416,1) en músculo de pescado contaminado a un tiempo de retención de 24,24 min, así como de 3 fragmentos de descomposición característicos con PM 336,2; 169,1 y 290,1 a 25,25 min, 22,45 min y 15,92 min, respectivamente. Además, se observa una variación en la abundancia relativa de los fragmentos en función del tipo de cocinado. De manera general, las técnicas de cocinado que conllevaban el uso de agua (especialmente el hervido), mostraron una abundancia relativa menor tanto de CYN, como de los distintos fragmentos, lo que parece indicar una pérdida de la toxina a través del agua del cocinado.Ministerio de Economía y Competitividad (AGL2015-64558-R y CGL2016-78937-R, MINECO/FEDER, UE).N

Analytical Pyrolysis of fish (Oreochromis niloticus) muscle. Effect of different cooking method

Póster (P-FA-34 ) presentado en la XVIII Reunión de la Sociedad Española de Cromatografía y Técnicas Afines (SECyTA 2018), Granada, del 2 al 4 de Octubre de 2018.In this communication a detailed analytical pyrolysis of tilapia fish (Oreochromis niloticus) muscle is described. The fish, supplied by Valenciana de Acuicultura (fish hatchery of Valencia, Spain), were acclimatized in the laboratory and for 15 days in two aquariums (8 individuals/aquarium) with 96 L of tap-water at a constant temperature (21 ± 2°C). Fish were fed daily (0.3 g/day) with commercial fish food only (Dibaq S.L., Segovia, Spain). After acclimation, were dissected and each muscle sample was cut into approximately 4 g portions. Fish muscle samples were cooked for 2 min by boiling, steaming, microwaving and broiling. Briefly, for boiling and steaming, the fish muscle was introduced into the pot or onto the food steamer, respectively, with cool water, heated to boiling (100°C) and continued to boil for 2 min. A conventional household microwave oven (Samsung M17-13, 300W, 2450 MHz) was used for microwaving, and samples were broiled in Teflon pans for both sides of the fillet. A non-cooked fish muscle fillet was used as control group. The assays were always carried out by quintuplicate (n=5). All samples were kept at -80°C until freeze dry (Cryodos 80, Telstar, Tarrasa, Spain). The Py-GC/MS was performed using a double-shot pyrolyzer (Frontier Laboratories, model 2020i, Fukushima, Japan) attached to a GC system (Agilent Technologies, Palo Alto, CA,. USA, model 6890N). The muscle samples (2 mg freeze-dry tissue) were placed in crucible deactivated steel pyrolysis capsules and introduced into a preheated micro-furnace at (350 °C) for 1 min. The volatile pyrolysates were then directly injected into the GC/MS for analysis. The gas chromatograph was equipped with a low polar-fused silica (5%-phenyl-methylpolysiloxane) capillary column (Agilent J&W HP-5ms Ultra Inert, of 30 m × 250 μm × 0.25 μm film thickness. The oven temperature was held at 50 °C for 1 min and then increased to 100 °C at 30 °C min-1, from 100 °C to 300 °C at 10 °C min-1, and stabilized at 300 °C for 10 min, with a total analysis time of 32 min. The carrier gas was He at a controlled flow of 1 mL min-1. The detector consisted of a mass selective detector (Agilent Technologies, Palo Alto, CA. USA, model 5973N) and mass spectra were acquired at 70 eV ionizing energy. Compound assignment was achieved by single-ion monitoring (SIM) for the major homologous series and by comparison with published data reported in the literature or stored in digital NIST 14 (Maryland, USA) and Wiley 7 (Weinheim, Germany) libraries. In a first analytical step, a detailed pyrolysis fingerprint of raw fish muscle tissue is produced and the effect of the pyrolysis temperature from 150 to 550 ºC in 100 ºC increments is studied in both, i) applying each temperature to a different sample or ii) sequentially (multi-shot) applying each temperature to the same sample. In a second phase, after stablishing an optimum pyrolysis temperature of 350 ºC for 1 minute, the effect of the different cooking methods (boil, steam,microwave and broil cooking) in the fish muscle pyrolyzates was studied.Projects CGL2016-78937-R and AGL2015-64558-R co-financed by FEDER Funds. Desiré Monis and Alba Carmona for technical assistance.N

Effects of Chrysosporum (Aphanizomenon) ovalisporum extracts containing cylindrospermopsin on growth, photosynthetic capacity, and mineral content of carrots (Daucus carota)

Natural toxins produced by freshwater cyanobacteria, such as cylindrospermopsin, have been regarded as an emergent environmental threat. Despite the risks for food safety, the impact of these water contaminants in agriculture is not yet fully understood. Carrots (Daucus carota) are root vegetables, extensively consumed worldwide with great importance for human nourishment and economy. It is, therefore, important to evaluate the possible effects of using water contaminated with cyanotoxins on carrot cultivation. The aim of this work was to investigate cylindrospermopsin effects on D. carota grown in soil and irrigated for 30 days, with a Chrysosporum ovalisporum extract containing environmentally relevant concentrations of cylindrospermopsin (10 and 50 μg/L). The parameters evaluated were plant growth, photosynthetic capacity, and nutritional value (mineral content) in roots of carrots, as these are the edible parts of this plant crop. The results show that, exposure to cylindrospermopsin did not have a clear negative effect on growth or photosynthesis of D. carota, even leading to an increase of both parameters. However, alterations in mineral contents were detected after exposure to crude extracts of C. ovalisporum containing cylindrospermopsin. A general decline was observed for most minerals (Ca, Mg, Na, Fe, Mn, Zn, Mo, and P), although an increase was shown in the case of K and Cu, pointing to a possible interference of the cyanobacterial extract in mineral uptake. This study is the first to evaluate the effects of C. ovalisporum extracts on a root vegetable, however, more research is necessary to understand the effects of this toxin in environmentally relevant scenarios. © 2016, Springer Science+Business Media New York.This work was partially funded by Funda????o para a Ci??ncia e Tecnologia (FCT), under the framework of the project UID/Multi/04423/2013, by Ministerio de Ciencia e Innovaci??n of Spain under the project AGL2009-10026, and by Junta de Andaluc??a under the project P09-AGR-4672. Remedios Guzm??n's work is supported by the Spanish Ministerio de Educaci??n fellowship. Alexandre Campos work is supported by a post-doctoral grant (SFRH/BPD/103683/2014) from FCT