Ready Access to Proquinazid Haptens via Cross-Coupling Chemistry for Antibody Generation and Immunoassay Development

17 pages, 4 tables, 5 figures.-- Publisher's PDFBioconjugate preparation is a fundamental step for antibody generation and immunoassay development to small chemical compounds. For analytical targets holding in their structure an aryl halogen atom, cross-coupling reactions may be a simple and efficient way to obtain functionalized derivatives; thus offering great potential to elicit robust and selective immune responses after being coupled to immunogenic carrier proteins. However, substitution of the halogen atom by an aliphatic chain might eventually compromise the affinity and specificity of the resulting antibodies. In order to address this issue, proquinazid, a new-generation fungicide with outstanding performance, was chosen as model analyte. Two functionalized derivatives differing in spacer arm rigidity were synthesized by Sonogashira cross-coupling chemistry. These haptens were covalently coupled to bovine serum albumin and the resulting immunoconjugates were employed for rabbit vaccination. Antibodies were tested for proquinazid recognition by direct and indirect competitive immunoassay, and IC50 values in the low nanomolar range were found, thus demonstrating the suitability of this straightforward synthetic strategy for the generation of immunoreagents to compounds bearing an aryl halide. Following antibody characterization, competitive immunoassays were developed and employed to determine proquinazid residues in grape musts, and their analytical performance was satisfactorily validated by comparison with GC–MS. Besides having described the development of the first immunochemical method for proquinazid analysis, an efficient functionalization approach for analytes comprising aryl halides is reported.Peer reviewe

Optimizing strategies for meningococcal C disease vaccination in Valencia (Spain)

BackgroundMeningococcal C (MenC) conjugate vaccines have controlled invasive diseases associated with this serogroup in countries where they are included in National Immunization Programs and also in an extensive catch-up program involving subjects up to 20 years of age. Catch-up was important, not only because it prevented disease in adolescents and young adults at risk, but also because it decreased transmission of the bacteria, since it was in this age group where the organism was circulating. Our objective is to develop a new vaccination schedule to achieve maximum seroprotection in these groups.MethodsA recent study has provided detailed age-structured information on the seroprotection levels against MenC in Valencia (Spain), where vaccination is routinely scheduled at 2 months and 6 months, with a booster dose at 18 months of age. A complementary catch-up campaign was also carried out in n for children from 12 months to 19 years of age. Statistical analyses of these data have provided an accurate picture on the evolution of seroprotection in the last few years.ResultsAn agent-based model has been developed to study the future evolution of the seroprotection histogram. We have shown that the optimum strategy for achieving high protection levels in all infants, toddlers and adolescents is a change to a 2 months, 12 months and 12 years of age vaccination pattern. If the new schedule were implemented in January 2014, high-risk subjects between 15-19 years of age would have very low seroprotection for the next 6 years, thereby threatening the program.ConclusionsHigh protection levels and a low incidence of meningococcal C disease can be achieved in the future by means of a cost-free change in vaccination program. However, we recommend a new catch-up program simultaneous to the change in regular vaccination program

Fermentación in vitro de pepsina/pancreatina con inóculos de conejos

The in vitro gas production of pepsin/pancreatin used in the determination of in vitro digestibility was evaluated using different inocula from rabbits (ileal, caecal and soft faeces from rabbits). In experiment 1 were used 3 different ileal and caecal inocula obtained each one from the combination of the digesta of 3 different 70 d old rabbits

Micro-mechanical response of ultrafine grain and nanocrystalline tantalum

In order to investigate the effect of grain boundaries on the mechanical response in the micrometer and submicrometer levels, complementary experiments and molecular dynamics simulations were conducted on a model bcc metal, tantalum. Microscale pillar experiments (diameters of 1 and 2 μm) with a grain size of ~100-200 nm revealed a mechanical response characterized by a yield stress of ~1500 MPa. The hardening of the structure is reflected in the increase in the flow stress to 1700 MPa at a strain of ~0.35. Molecular dynamics simulations were conducted for nanocrystalline tantalum with grain sizes in the range of 20-50 nm and pillar diameters in the same range. The yield stress was approximately 6000 MPa for all specimens and the maximum of the stress-strain curves occurred at a strain of 0.07. Beyond that strain, the material softened because of its inability to store dislocations. The experimental results did not show a significant size dependence of yield stress on pillar diameter (equal to 1 and 2 um), which is attributed to the high ratio between pillar diameter and grain size (~10-20). This behavior is quite different from that in monocrystalline specimens where dislocation 'starvation' leads to a significant size dependence of strength. The ultrafine grains exhibit clear 'pancaking' upon being plastically deformed, with an increase in dislocation density. The plastic deformation is much more localized for the single crystals than for the nanocrystalline specimens, an observation made in both modeling and experiments. In the molecular dynamics simulations, the ratio of pillar diameter (20-50 nm) to grain size was in the range 0.2-2, and a much greater dependence of yield stress to pillar diameter was observed. A critical result from this work is the demonstration that the important parameter in establishing the overall deformation is the ratio between the grain size and pillar diameter; it governs the deformation mode, as well as surface sources and sinks, which are only important when the grain size is of the same order as the pillar diameter.Fil: Yang, Wen. University of California at San Diego; Estados UnidosFil: Ruestes, Carlos Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto Interdisciplinario de Ciencias Básicas. - Universidad Nacional de Cuyo. Instituto Interdisciplinario de Ciencias Básicas; ArgentinaFil: Li, Zezhou. University of California at San Diego; Estados UnidosFil: Abad, Oscar Torrents. Leibniz Institute for New Materials; AlemaniaFil: Langdon, Terence G.. University of Southern California; Estados UnidosFil: Heiland, Birgit. Leibniz Institute for New Materials; AlemaniaFil: Koch, Marcus. Leibniz Institute for New Materials; AlemaniaFil: Arzt, Eduard. Leibniz Institute for New Materials; Alemania. Universitat Saarland; AlemaniaFil: Meyers, Marc A.. University of California at San Diego; Estados Unido

Efecto de la suplementación con Bacillus amyloliquefaciens y heptanoato sódico sobre los parámetros de crecimiento en conejos en cebo

El objetivo de este trabajo fue evaluar el efecto de suplementar con B. amyloliquefaciens (Ecobiol; EU aditivo zootécnico para piensos número 4b1822; contenido de 1 x 109 CFU de B. amyloliquefaciens CECT 5940 / g) y heptanoato sódico (Hepton) sobre los parámetros de crecimiento del conejo. Se formuló un pienso control (17,3% PB y 36,2% FND, sobre % MS), y otros tres piensos suplementando el pienso control con 500 y 1000 g B. amyloliquefaciens/t (BA1 y BA2) y con 1000 g heptanoato sódico/t. Se utilizaron gazapos destetados a los 25 d gazapos (67/pienso). Los conejos suplementados con el pienso BA2 tendieron a comer menos que el grupo control (P = 0,073), y comieron un 6% menos que los del grupo BA1 (P = 0,036) durante el periodo de 25 a 40 d de edad, si bien no afectó a la ganancia de peso, ni en la eficacia alimenticia. Durante el periodo de 40 a 63 d de edad los gazapos alimentados con el pienso BA2 comieron un 10% menos que el pienso control (P = 0,002), sin modificar la ganancia de peso, lo que se tradujo en una tendencia a mejorar la eficacia alimenticia (P = 0,063). Los animales del pienso BA2 incrementaron la eficacia alimenticia un 8% comparado con los del grupo BA1 durante el periodo de 40 a 63 d (P = 0,031), sin mostrar diferencias en el consumo o en la ganancia de peso. Al evaluar el periodo global, incrementar la dosis de B. amyloliquefaciens (BA2 vs. BA1) tendió a disminuir el consumo medio diario (P = 0,069) y a aumentar la eficiencia alimenticia un 6% (P = 0,006). La suplementación con B. amyloliquefaciens, independientemente de la dosis utilizada (BA2 o BA1), no afectó a la mortalidad en el periodo global del cebo. Durante todo el periodo experimental de 25 a 63 d de edad, la suplementación con heptanoato sódico redujo el consumo de pienso un 5% en comparación con el grupo control (P = 0,050) y, al no modificar la velocidad de crecimiento, aumentó la eficacia alimenticia un 7% (P = 0,003). En comparación con el promedio de los gazapos suplementados con B. amyloliquefaciens los conejos que recibieron heptanoato sódico mostraron una mayor ganancia de peso (5%, P = 0,012), y eficiencia alimenticia (4%, P = 0,024), alcanzando un peso final superior (P = 0,012)

Effect of sugar beet pulp fibre fractions on growth performance, fecal digestibility and digestive physiology in rabbits around weaning

The present study investigated the effect of the different fibre components of sugar beet pulp (SBP) on growth performance and some digestive traits. Four semi-synthetic diets were formulated with similar NDF (33% DM) and protein (16% DM) level. Control diet was formulated to contain the lowest level of soluble fibre (3% DM) and SBP diet the highest (9%). The soluble (pectins) and insoluble fractions of SBP were studied in other two diets (Pectin and InsSBP diets). A total of 136 weanling rabbits (25 d of age) was housed individually, randomly assigned to 4 experimental groups, and fed ad libitum with the experimental diets during 10 days after weaning. The type of diet did not affect growth rate and stomach pH. Animals fed with SBP diet showed higher DM and NDF digestibility (4 and 83%, respectively), gain:feed ratio (13%), cecal and total tract weight (13 and 9%) and ileal viscosity (148%) than rabbits fed the Control diet, but lower cecal pH (9%). Pectin diet increased ileal viscosity and decreased the weight of stomach content with respect to SBP diet. Rabbits fed InsSBP diet showed similar results to SBP diet but lower ileal viscosity and cecal pH than those fed Pectin diet. In conclusion, SBP and their soluble and insoluble fractions are well digested in young rabbits. However the soluble and insoluble fibre of SBP produce different effects in the gastrointestinal tract

Toward a clinical practice guide in pharmacogenomics testing for functional polymorphisms of drug-metabolizing enzymes. Gene/drug pairs and barriers perceived in Spain

The development of clinica lpractice recommendations or guidelines for the clinical use of biomarkers is an issue of great importance withr regard to adverse drug reactions.The poten-tial of pharmacogenomicbiomarkers has been extensively investigated in recent years.However,several barriers to implementing the use of pharmacogenomics testing exist.We conducted a survey among members of the Spanish Societies of Pharmacology and Clinical Pharmacology to obtain information about the perception of such barriers and to compare the perceptions of participants about the relative importance of majorgene/drug pairs.Of 11 potential barriers,the highest importance was attributed to lack of institutional support for pharmacogenomic stesting,and to the issues related to the lack of guidelines.Of the proposed gene/drug pairs the highest importance was assigned to HLA-B/abacavir, UGT1A1/irinotecan, and CYP2D6/tamoxifen.In this perspective article,we compare the relative importance of 29 gene/drugpairs in the Spanish study with that of the same pairs in the American Society for Clinical Pharmacology and Therapeutic sstudy,and we provide suggestions and areas of focus to develop a guide for clinical practice in pharmacogenomics testingThe work in the author’s laboratory is financed by Grants PS09/00943, PS09/00469, RETICS RIRAAF RD07/0064/0016, and CIBERehd from Instituto de Salud CarlosIII,Madrid, Spain, and by Grants GR10068 from Junta de Extremadura, Spain. Financed in part with FEDER funds from the European Unio

Mapping the habitats of a complex circalittoral rocky shelf in the Cantabrian Sea (south Bay of Biscay)

This work focuses on the study of habitats and communities of a high structural complexity area at different levels and scales. This gives us a better understanding of an area from an ecological point of view and at the same time provides us with tools that will facilitate management measures. It was developed in a complex circalittoral rocky platform of the central Cantabrian Sea (Bay of Biscay). The sampling was carried out using a towed photogrammetric vehicle and a rock dredge, which was used for the identification of the species. The first level of the study was the abiotic characterization of the area and the analysis of the communities. These analysis were developed using the unsupervised k-means classification method. For abiotic characterization we used the variables directly associated with the composition and morphology of the ground, such as backscatter, BPI (Bathymetric Position Index), roughness and slope. Depth was also included to discriminate between the circalittoral and bathyal zones. We obtained 5 different classes, which we related to the ground types observed by photogrammetry. In the analysis of the communities, the cluster was based on the sampling units extracted from the images (~10 m), from which 5 assemblages were obtained, providing information on the most abundant species of each class supplied by the abiotic study. The second level was carried out considering a management approach and was based on the modeling of the area at lower resolution, more suitable for the analysis of the habitat-fisheries interactions. Thus, the main habitat-forming species (HFS) of the entire circalittoral area were used to perform delta models based on GAMs (Generalize Additive Models). Obtaining the predictions of presence/absence and combining it with the predictions of densities, we got the zero inflated values density-based model. As all the identified habitats have vulnerable benthic species of a certain size settled on rocky bottoms, they can all be considered to belong to the designation 1170 reefs of the Habitats Directive

Prevalence estimation of significant fibrosis because of NASH in Spain combining transient elastography and histology

Acord transformatiu CRUE-CSICBackground & Aims: Non-alcoholic fatty liver disease (NAFLD) has become a major public health problem, but the prevalence of fibrosis associated with non-alcoholic steatohepatitis (NASH) is largely unknown in the general population. This study aimed to provide an updated estimation of the prevalence of NASH fibrosis in Spain. Methods: This was an observational, retrospective, cross-sectional, population-based study with merged data from two Spanish datasets: a large (N = 12 246) population-based cohort (ETHON), including transient elastography (TE) data, and a contemporary multi-centric biopsy-proven NASH cohort with paired TE data from tertiary centres (N = 501). Prevalence for each NASH fibrosis stage was estimated by crossing TE data from ETHON dataset with histology data from the biopsy-proven cohort. Results: From the patients with valid TE in ETHON dataset (N = 11 440), 5.61% (95% confidence interval [95% CI]: 2.53-11.97) had a liver stiffness measurement (LSM) ≥ 8 kPa. The proportion attributable to NAFLD (using clinical variables and Controlled Attenuation Parameter) was 57.3% and thus, the estimated prevalence of population with LSM ≥ 8 kPa because of NAFLD was 3.21% (95% CI 1.13-8.75). In the biopsy-proven NASH cohort, 389 patients had LSM ≥ 8 kPa. Among these, 37% did not have significant fibrosis (F2-4). The estimated prevalence of NASH F2-3 and cirrhosis in Spain's adult population were 1.33% (95% CI 0.29-5.98) and 0.70% (95% CI 0.10-4.95) respectively. Conclusions: These estimations provide an accurate picture of the current prevalence of NASH-related fibrosis in Spain and can serve as reference point for dimensioning the therapeutic efforts that will be required as NASH therapies become available