Transversal gene expression panel to evaluate intestinal health in broiler chickens in different challenging conditions.

There is a high interest on gut health in poultry with special focus on consequences of the intestinal diseases, such as coccidiosis and C. perfringens-induced necrotic enteritis (NE). We developed a custom gene expression panel, which could provide a snapshot of gene expression variation under challenging conditions. Ileum gene expression studies were performed through high throughput reverse transcription quantitative real-time polymerase chain reaction. A deep review on the bibliography was done and genes related to intestinal health were selected for barrier function, immune response, oxidation, digestive hormones, nutrient transport, and metabolism. The panel was firstly tested by using a nutritional/Clostridium perfringens model of intestinal barrier failure (induced using commercial reused litter and wheat-based diets without exogenous supplementation of enzymes) and the consistency of results was evaluated by another experiment under a coccidiosis challenge (orally gavaged with a commercial coccidiosis vaccine, 90× vaccine dose). Growth traits and intestinal morphological analysis were performed to check the gut barrier failure occurrence. Results of ileum gene expression showed a higher expression in genes involved in barrier function and nutrient transport in chickens raised in healthy conditions, while genes involved in immune response presented higher expression in C.perfringens-challenged birds. On the other hand, the Eimeria challenge also altered the expression of genes related to barrier function and metabolism, and increased the expression of genes related to immune response and oxidative stress. The panel developed in the current study gives us an overview of genes and pathways involved in broiler response to pathogen challenge. It also allows us to deep into the study of differences in gene expression pattern and magnitude of responses under either a coccidial vaccine or a NE.

Dietary ß-galactomannans have beneficial effects on the intestinal morphology of chickens challenged with Salmonella enterica serovar Enteritidis.

Salmonella enterica serovar Enteritidis is one of the leading causes of food-borne salmonellosis in humans. Poultry is the single largest reservoir, and the consumption of incorrectly processed chicken meat and egg products is the major source of infection. Since 2006, the use of antibiotics as growth promoters has been banned in the European Union, and the dietary inclusion of ß-galactomannans (ßGM) has become a promising strategy to control and prevent intestinal infections. The aim of this study was to investigate the effect of various ßGM-rich products on intestinal morphology in chickens challenged with Salmonella Enteritidis. To assess this effect, a total of 280 male Ross 308 chickens were studied (40 animals per treatment housed in 5 cages). There were 7 treatments, including controls: uninoculated birds fed the basal diet (negative control) and inoculated birds fed the basal diet (positive control) or the basal diet supplemented with Salmosan (1 g/kg), Duraió gum (1 g/kg), Cassia gum (1 g/kg), the cell walls of Saccharomyces cerevisiae (0.5 g/kg), or the antibiotic colistine (0.8 g/kg). The birds were fed these diets from the d 1 to 23, except the animals in the colistine group, which were fed the diet containing the antibiotic only from d 5 to 11. The inoculated animals were orally infected on d 7 with 10(8) cfu of Salmonella Enteritidis. Bird performance per replicate was determined for the whole study period (23 d), and the distal ileum and cecal tonsil of 5 animals per treatment (1 animal per replicate) were observed at different magnification levels (scanning electron, light, and laser confocal microscopy). In the images corresponding to the treatments containing ßGM we observed more mucus, an effect that can be associated with the observation of more goblet cells. Moreover, the images also show fewer M cells, which are characteristic of infected animals. Regarding the morphometric parameters, the animals that received Duraió and Cassia gums show greater (P = 0.001 and P = 0.016, respectively) villus length compared with the animals in the positive control, thus indicating the capacity of these products to increase epithelial surface area. However, no effect (P > 0.05) on microvillus dimensions was detected. In conclusion, the results obtained indicating the beneficial effects of these ßGM on intestinal morphology give more evidence of the positive effects of these supplements in poultry nutrition.

Establishment of an in vitro photoassay using THP-1 cells and IL-8 to discriminate photoirritants from photoallergens.

At present, there are no in vivo or in vitro methods developed which has been adopted by regulatory authorities to assess photosensitization induced by chemicals. Recently, we have proposed the use of THP-1 cells and IL-8 release to identify the potential of chemicals to induce skin sensitization. Based on the assumption that sensitization and photosensitization share common mechanisms, the aim of this work was to explore the THP-1 model as an in vitro model to identify photoallergenic chemicals. THP-1 cells were exposed to 7 photoallergens and 3 photoirritants and irradiated with UVA light or kept in dark. Non phototoxic allergens or irritants were also included as negative compounds. Following 24h of incubation, cytotoxicity and IL-8 release were measured. At subtoxic concentrations, photoallergens produced a dose-related increase in IL-8 release after irradiation. Some photoirritants also produced a slight increase in IL-8 release. However, when the overall stimulation indexes of IL-8 were calculated for each chemical, 6 out of 7 photoallergens tested reached a stimulation index above 2, while the entire set of negative compounds had stimulation indexes below 2. Our data suggest that this assay may become a useful cell-based in vitro test for evaluating the photosensitizing potential of chemicals.

[Viscosity changes in thickened water due to the addition of highly prescribed drugs in geriatrics]. / Cambios en la viscosidad del agua con espesantes por la adición de fármacos altamente prescritos en geriatría.

INTRODUCTION: Dysphagia is a swallowing disorder with a high incidence in the geriatric patient related with an increased risk for undernutrition and pneumonia due to bronchial aspiration. In this condition, it is usual to add commercial thickeners in liquids, as well as the addition of drugs in this mixture to improve their administration. However, there are no studies regarding the possible change in viscosity produced by their addition. OBJECTIVES: To assess the change in viscosity of water thickened with commercial products by adding the drugs frequently used in elderly patients. METHODS: Samples of water mixed with the commercial thickener Resource (modified corn starch) or Nutilis (modified corn starch, maltodextrin, and gums: tara, xhantan, and guar) to achieve an intermediate consistence as "honey". The viscosity of these samples was measured as well as for similar samples to which one of the following drugs was added: galantamine, rivastigmin, ciprofloxacin, cholecalciferol, memantine, fosfomycin, calcium, and amoxicillin/clavulanic acid. RESULTS: In the samples with Resource thickener we observed decreased viscosity by adding galantamine, memantine, fosfomycin or calcium, and increased viscosity with amoxicillin/clavulanic acid. The viscosity of the samples with Nutilis® decreased with galantamine, rivastigmine, amoxicillin/clavulanic acid, fosfomycin and calcium. CONCLUSION: The viscosity of water with commercial thickeners may be affected by some drugs or their preservatives, which may influence the swallowing capability. It is recommended to perform further in vitro and in vivo studies in order to adjust these formulations if necessary.

Cambios en la viscosidad del agua con espesantes por la adición de fármacos altamente prescritos en geriatría / Viscosity changes in thickened water due to the addition of highly prescribed drugs in geriatrics

Introducción: La disfagia es una alteración de la deglución con una elevada incidencia en el paciente geriátrico relacionada con un aumento del riesgo de desnutrición y neumonía por broncoaspiración. La adición de espesantes comerciales en líquidos es frecuente en esta situación, así como la adición de fármacos en esta mezcla para facilitar su administración. Sin embargo, no existen estudios referentes al posible cambio de viscosidad producido por la adición de los mismos. Objetivos: Evaluar la variación ejercida sobre la viscosidad del agua espesada con preparados comerciales al añadir fármacos frecuentemente utilizados en pacientes de edad avanzada. Métodos: Se prepararon muestras de agua con espesante comercial Resource® (almidón de maíz modificado) o Nutilis® (almidón modificado de maíz, maltodextrina y gomas: tara, xantana y guar) para conseguir consistencia intermedia tipo "miel". Se midió la viscosidad de estas muestras y para muestras similares a las que se había añadido alguno de los siguientes fármacos: galantamina, rivastigmina, ciprofloxacino, colecalciferol, memantina, fosfomicina, calcio y amoxilina/clavulánico. Resultados: En las muestras con espesante Resource® se observó una disminución de la viscosidad al añadir galantamina, memantina, fosfomicina o calcio, y un aumento de la viscosidad con amoxicilina/clavulánico. La viscosidad de la muestras con Nutilis® disminuyó con galantamina, rivastigmina, amoxicilina/clavulánico, fosfomicina y calcio. Conclusión: La viscosidad del agua con espesantes comerciales puede verse afectada por algunos fármacos o sus excipientes, lo que puede incidir en la capacidad de deglución de los mismos. Es aconsejable realizar más estudios in vitro e in vivo para valorar ajustar dichas pautas en caso necesario (AU)

Introduction: Dysphagia is a swallowing disorder with a high incidence in the geriatric patient related with an increased risk for undernutrition and pneumonia due to bronchial aspiration. In this condition, it is usual to add commercial thickeners in liquids, as well as the addition of drugs in this mixture to improve their administration. However, there are no studies regarding the possible change in viscosity produced by their addition. Objectives: To assess the change in viscosity of water thickened with commercial products by adding the drugs frequently used in elderly patients. Methods: Samples of water mixed with the commercial thickener Resource® (modified corn starch) or Nutilis® (modified corn starch, maltodextrin, and gums: tara, xhantan, and guar) to achieve an intermediate consistence as "honey". The viscosity of these samples was measured as well as for similar samples to which one of the following drugs was added: galantamine, rivastigmin, ciprofloxacin, cholecalciferol, memantine, fosfomycin, calcium, and amoxicillin/clavulanic acid. Results: In the samples with Resource® thickener we observed decreased viscosity by adding galantamine, memantine, fosfomycin or calcium, and increased viscosity with amoxicillin/clavulanic acid. The viscosity of the samples with Nutilis® decreased with galantamine, rivastigmine, amoxicillin/clavulanic acid, fosfomycin and calcium. Conclusion: The viscosity of water with commercial thickeners may be affected by some drugs or their preservatives, which may influence the swallowing capability. It is recommended to perform further in vitro and in vivo studies in order to adjust these formulations if necessary (AU)

Moodleando en posgrado: uso del campus virtual y Evaluación continuada / Moodle at prosgraduate level: virtual platform utilization and continuous assesment

La adaptación al EEES ha implicado una reestructuración de los estudios universitarios promoviendo, entre otros, el uso de una evaluación continuada de los aprendizajes y de herramientas docentes virtuales en los nuevos grados. Estos cambios también pueden aplicarse en el posgrado, aunque hasta el momento, son pocas las experiencias al respecto. El presente trabajo muestra la opinión y los resultados obtenidos de los estudiantes de una asignatura de posgrado de un Máster Oficial Universitario URV-UB en el que se llevó a cabo evaluación continuada y se utilizó el Campus Virtual (CV) como herramienta de aprendizaje y participación activa. La aplicación del proyecto (2007PID/UB-14) se llevó a cabo en 4 fases: elaboración del aula, diseño de encuestas dirigidas a los estudiantes para obtener su opinión sobre la evaluación continuada y sus conocimientos en el uso de plataformas virtuales de forma previa al curso, gestión diaria del aula y seguimiento de las actividades propuestas y finalmente, evaluación del proyecto a través de indicadores cualitativos y cuantitativos derivados de las encuestas y de la propia aplicación virtual. Al inicio del curso la mayoría (93,75%) de los estudiantes prefería evaluación continuada y al final de éste el 100% estuvieron de acuerdo con el tipo de evaluación realizado. Respecto al CV, los estudiantes realizaron todas las actividades de autoevaluación voluntarias. La puntuación sobre la utilidad de todos los recursos del aula del CV fue muy elevada, siendo máxima la obtenida por los ejercicios de autoevaluación. En global, los estudiantes otorgaron al aula una puntuación de 8,1 sobre 10(AU)

The definition of new academic degrees according to the EHEA has promoted, among others, the utilization of graduate students learning by continuous evaluation, as well as the usage of virtual teaching tools. These changes can also be applicable for postgraduate students, although very few experiences exist in this sense. The present work shows the students’ opinion and results from a URV-UB EHEA Master’s subject in which continuous assessment and virtual teaching tools for active learning and participation have been used. The project (2007PID/UB-14) was performed in 4 steps: virtual material production, surveys to students designed to obtain information regarding continuous assessment and their knowledge about virtual platforms (VP), daily assessment of proposed activities, and finally, evaluation of the project by means of qualitative and quantitative indicators from surveys and from the virtual platform usage profiles. At the beginning of the course most of students (93.75%) preferred continuous assessment and at the end 100% agreed with the type of evaluation applied. With respect to the VP, the students performed all the voluntary activities. All virtual teaching tools were considered as very useful by the students, being the most useful for them the autoevaluation activities(AU)

Elaboración de un video sobre calidad, seguridad y medio ambiente en la unidad de laboratorios docentes (ULD) de la Facultad de Farmacia de la Universidad de Barcelona / Development of a video on quality, safety and environment in the teaching laboratories unit (ULD) of the Faculty of Pharmacy of the University of Barcelona

La Unidad de Laboratorios Docentes (ULD) de la Facultad de Farmacia (UB) ha implantado unsistema de gestión de la calidad (SGC) que permite transmitir al estudiante una formación adicionalcon el objetivo de mejorar sus competencias transversales (siguiendo las directrices del EspacioEuropeo de Educación Superior, EEES). Además, en el curso académico 06-07, se instauraron lasBuenas Prácticas Ambientales (BPAL) para disponer de un sistema de gestión que incorporase loscriterios de calidad, medioambiente y seguridad (sistema de gestión integrada, SGI). Durante elpresente curso académico se ha procedido a la grabación y edición de un video sobre calidad,seguridad y medioambiente en la ULD con el objetivo de mejorar la formación transversal de losestudiantes facilitando la integración de los conocimientos y habilidades profesionales. De esta forma,al salir del entorno universitario, los licenciados y graduados de la Facultad de Farmacia dispondrán deun valor añadido en su formación, mejorando así sus competencias para el desarrollo de su futuraprofesión. En el video se muestra la manera de trabajar correctamente según las normas de calidad,seguridad y medioambiente recogidas además en un tríptico que se entrega a los estudiantes al accederpor primera vez a un laboratorio de prácticas. El video se difundirá a través de la página web de laULD, de la videoteca de la UB, del canal You Tube Canal UB, así como de las asignaturas que losoliciten (sirviendo de soporte para el personal docente)(AU)

The Teaching Laboratories Unit (ULD) of the Faculty of Pharmacy (UB) has implemented amanagement system (QMS) that allows the student to get an additional training in order to improvetheir generic skills (following the guidelines of the Area European Higher Education Area, EHEA).Furthermore, in the 2006-2007 academic year, were introduced the Good Laboratory Practice (GLP)to have a management system that incorporates the criteria of quality, environment and security(integrated management system, IMS). During the present academic year it has been carried out therecording and editing of a video on quality, safety and environment in the ULD with the aim ofimproving cross-training of students by facilitating the integration of knowledge and professionalskills. Thus, on leaving the university, graduates in Pharmacy will have an added value in theirtraining, and thereby improving their skills for the development of their future profession. The videoshows how to work correctly according to the standards of quality, safety and environment that arealso contained in a leaflet that is given to students when they first access to a laboratory. The video will be disseminated through the website of the ULD, the library of the UB, the You Tube UB Canaland upon request(AU)

Partial Na+ dependence of DL-2-hydroxy-4-(methylthio)butanoic acid uptake in the chicken small intestine.

Methionine hydroxy analogue DL-2-hydroxy-(4-methylthio)butanoic acid (HMTBA) is commonly used as a supplemental source of Met in commercial animal diets. To better understand the uptake of this analogue by the chicken intestine, the aim of the present study was to assess the contribution of Na+ gradient on HMTBA accumulation in everted sacs of the chicken small intestine (duodenum, jejunum, and ileum). In the presence of an H+ gradient, uptake was lower in the absence of Na+ along the chicken small intestine, although no significant differences were detected in the duodenum. In contrast, in the absence of an H+ gradient, no significant differences were detected between the 2 Na+ conditions. In conclusion, the observed relationship between Na+ and H+ dependence indicates the participation of the apical Na+/H+ exchanger in HMTBA uptake in the chicken small intestine.

Conversion of the methionine hydroxy analogue DL-2-hydroxy-(4-methylthio) butanoic acid to sulfur-containing amino acids in the chicken small intestine.

dl-Methionine or its corresponding hydroxy analogue, DL-2-hydroxy-(4-methylthio) butanoic acid (DLHMB), are commonly added to commercial animal diets to satisfy the TSAA requirement. The utilization of DLHMB as a supplementary source of Met begins with its conversion to L-Met via a 2-step mediated process. L-Methionine can then be transsulfurated to L-Cys, which, in turn, can be catabolized to taurine (TAU). In the present study, the capacity of the chicken small intestine to convert DLHMB to L-Met and to use this amino acid as a source for L-Cys and TAU production was evaluated. The appearance of Met in the serosal compartment of everted sacs incubated with DLHMB is higher in the presence of an H(+) gradient (mucosal pH 5.5 vs. 7.4). Serosal Cys and TAU concentration was compared in everted sacs incubated at a mucosal pH of 5.5 with DLHMB or L-Met, and the results show significantly higher values after incubation with the hydroxy analogue. Regional comparisons indicate no significant differences in the appearance of serosal Met and Cys, although lower values were obtained for TAU in the duodenum than in the jejunum and ileum. The profile of non-S amino acids was also determined and revealed no significant differences between DLHMB- and L-Met-incubated sacs. In conclusion, Cys and TAU content in chicken enterocytes is higher when DLHMB is used as a Met source.

Oligomers are not the limiting factor in the absorption of DL-2-hydroxy-4-(methylthio)butanoic acid in the chicken small intestine.

The methionine hydroxy analogue DL-2-hydroxy-4-(methylthio)butanoic acid (HMB) is commonly used as a supplemental source of methionine in commercial animal diets. The HMB free acid is an aqueous solution that contains 88% product in an equilibrium mixture of monomer, dimer, and polymeric compounds. The present study examines whether the presence of these nonmonomeric forms reduces the absorption of the hydroxy analogue in the chicken small intestine. In vivo and in vitro methodologies were used to compare the intestinal absorption of an HMB product containing mainly monomer (HMB-PCM) with commercial HMB. The results from the in vivo perfusion of the jejunum showed no significant differences between the 2 hydroxy analogue sources in monomer absorption from the intestinal lumen, tissue accumulation, or plasma concentration. The results also indicate that the nonmonomeric forms are hydrolyzed during perfusion. Moreover, monomer tissue accumulation in everted sacs showed no significant differences between substrates, either in the presence or in the absence of a H+-gradient; a higher value was observed in the jejunum and ileum in comparison with the duodenum. Similarly, serosal appearance in H+-gradient conditions did not differ significantly between substrates, and it showed the same regional profile as in tissue accumulation. Oligomer hydrolysis was confirmed in vitro without significant differences between segments. In conclusion, the presence of nonmonomeric forms is not a limiting factor in HMB absorption, apparently because of the hydrolytic capacity of intestinal mucosa, as confirmed by experiments in vivo and in vitro.