Simultaneous evaluation of multiple microarray surface chemistries through real-time interferometric imaging.

Surface chemistry is a crucial aspect for microarray modality biosensor development. The immobilization capability of the functionalized surface is indeed a limiting factor for the final yield of the binding reaction. In this work, we were able to simultaneously compare the functionality of protein ligands that were locally immobilized on different polymers, while on the same solid support, therefore demonstrating a new way of multiplexing. Our goal was to investigate, in a single experiment, both the immobilization efficiency of a group of reactive polymers and the resulting affinity of the tethered molecules. This idea was demonstrated by spotting many reactive polymers on a Si/SiO2 chip and depositing the molecular probes on the spots immediately after. As a proof of concept, we focused on which polymers would better immobilize a model protein (α-Lactalbumin) and a peptide (LAC-1). We successfully showed that this protocol is applicable to proteins and peptides with a good efficiency. By means of real-time binding measurements performed with the interferometric reflectance imaging sensor (IRIS), local functionalization proved to be comparable to the classical flat coating solution. The final outcome highlights the multiplexing power of this method: first, it allows to characterize dozens of polymers at once. Secondly, it removes the limitation, related to coated surfaces, that only molecules with the same functional groups can be tethered to the same solid support. By applying this protocol, many types of molecules can be studied simultaneously and immobilization for each probe can be individually optimized.766466 (INDEX) - Horizon 2020 Framework Programmehttps://s3-eu-west-1.amazonaws.com/itempdf74155353254prod/8976347/Simultaneous_Evaluation_of_Multiple_Microarray_Surface_Chemistries_Through_Real-Time_Interferometric_Imaging_v1.pdfFirst author draf

Safety and efficacy of cognitive training plus epigallocatechin-3-gallate in young adults with Down's syndrome (TESDAD): a double-blind, randomised, placebo-controlled, phase 2 trial

Background Early cognitive intervention is the only routine therapeutic approach used for amelioration of intellectual deficits in individuals with Down's syndrome, but its effects are limited. We hypothesised that administration of a green tea extract containing epigallocatechin-3-gallate (EGCG) would improve the effects of non-pharmacological cognitive rehabilitation in young adults with Down's syndrome.; Methods We enrolled adults (aged 16-34 years) with Down's syndrome from outpatient settings in Catalonia, Spain, with any of the Down's syndrome genetic variations (trisomy 21, partial trisomy, mosaic, or translocation) in a double-blind, placebo-controlled, phase 2, single centre trial (TESDAD). Participants were randomly assigned at the IMIM-Hospital del Mar Medical Research Institute to receive EGCG (9 mg/kg per day) or placebo and cognitive training for 12 months. We followed up participants for 6 months after treatment discontinuation. We randomly assigned participants using random-number tables and balanced allocation by sex and intellectual quotient. Participants, families, and researchers assessing the participants were masked to treatment allocation. The primary endpoint was cognitive improvement assessed by neuropsychologists with a battery of cognitive tests for episodic memory, executive function, and functional measurements. Analysis was on an intention-to-treat basis. This trial is registered with ClinicalTrials.gov, number NCT01699711.; Findings The study was done between June 5, 2012, and June 6, 2014. 84 of 87 participants with Down's syndrome were included in the intention-to-treat analysis at 12 months (43 in the EGCG and cognitive training group and 41 in the placebo and cognitive training group). Differences between the groups were not significant on 13 of 15 tests in the TESDAD battery and eight of nine adaptive skills in the Adaptive Behavior Assessment System II (ABAS-II). At 12 months, participants treated with EGCG and cognitive training had significantly higher scores in visual recognition memory (Pattern Recognition Memory test immediate recall, adjusted mean difference: 6.23 percentage points [95% CI 0.31 to 12.14], p=0.039; d 0.4 [0.05 to 0.84]), inhibitory control (Cats and Dogs total score, adjusted mean difference: 0.48 [0.02 to 0.93], p=0.041; d 0.28 [0.19 to 0.74]; Cats and Dogs total response time, adjusted mean difference: -4.58 s [-8.54 to -0.62], p=0.024; d -0.27 [-0.72 to -0.20]), and adaptive behaviour (ABAS-II functional academics score, adjusted mean difference: 5.49 [2.13 to 8.86], p=0.002; d 0.39 [-0.06 to 0.84]). No differences were noted in adverse effects between the two treatment groups.; Interpretation EGCG and cognitive training for 12 months was significantly more effective than placebo and cognitive training at improving visual recognition memory, inhibitory control, and adaptive behaviour. Phase 3 trials with a larger population of individuals with Down's syndrome will be needed to assess and confirm the long-term efficacy of EGCG and cognitive training.; Funding Jerome Lejeune Foundation, Instituto de Salud Carlos III FEDER, MINECO, Generalitat de Catalunya.Peer ReviewedPostprint (author's final draft

Impact of Spontaneous Fermentation and Inoculum with Natural Whey Starter on Peptidomic Profile and Biological Activities of Cheese Whey: A Comparative Study

Fermentation is a promising solution to valorize cheese whey, the main by-product of the dairy industry. In Parmigiano Reggiano cheese production, natural whey starter (NWS), an undefined community of thermophilic lactic acid bacteria, is obtained from the previous day residual whey through incubation at gradually decreasing temperature after curd cooking. The aim of this study was to investigate the effect of fermentation regime (spontaneous (S) and NWS-inoculated (I-NWS)) on biofunctionalities and release of bioactive peptides during whey fermentation. In S and I-NWS trials proteolysis reached a peak after 24 h, which corresponded to the drop out in pH and the maximum increase in lactic acid. Biological activities increased as a function of fermentation time. NWS inoculum positively affected antioxidant activity, whilst S overcame I-NWS in angiotensin-converting enzyme (ACE) and DPP-IV (dipeptidyl peptidase IV) inhibitory activities. Peptidomics revealed more than 400 peptides, mainly derived from β-casein, κ-casein, and α-lactalbumin. Among them, 49 were bioactive and 21 were ACE-inhibitors. Semi-quantitative analysis strongly correlated ACE-inhibitory activity with the sum of the peptide abundance of ACE-inhibitory peptides. In both samples, lactotripeptide isoleucine-proline-proline (IPP) was higher than valine-proline-proline (VPP), with the highest content in S after 24 h of fermentation. In conclusion, we demonstrated the ability of whey endogenous microbiota and NWS to extensively hydrolyze whey proteins, promoting the release of bioactive peptides and improving protein digestibility

Size matters: Physiological sensitivity of the scallop Argopecten purpuratus to seasonal cooling and deoxygenation upwelling-driven events

Environment imposes physiological constraints which are life-stage specific as growth-maintenance and/or growth-reproduction energetic requirements are size and volume-dependent. The scallop Argopecten purpuratus, one of the most important bivalve species subjected to fishery and aquaculture along the Humboldt Current System, inhabits spaces affected by continuous changes in temperature, pH, oxygen, and food availability driven by remote and local oceanographic processes. Specifically, in Chile, this species is mainly cultured in central-north Chile where is permanently affected by upwelling events of dissimilar intensity and duration which generate local conditions of acidification, deoxygenation, and cooling with different magnitudes. However, to date, it remains unknown how this economic valuable resource is physiologically affected throughout its life cycle by the continuous environmental changes driven by upwelling events of different intensities and duration along the year. Here, for the first time, A. purpuratus life-stage physiological sensitivity was assessed at a seasonal scale through a year-field experiment where growth, calcification, and survivorship were evaluated. Our study shows how seasonal differences in the upwelling phenology (here measured as changes in temperature, dissolved oxygen, pH, and primary productivity, but also as the number, duration, and intensity of cooling and de-oxygenation events) notably impacted the A. purpuratus physiological performance from juvenile to adult life-stages. This was especially noticeable during the spring season which showed the most intense cooling and deoxygenation events driven by stronger favorable-upwelling winds and the lowest growth and gross calcification rates (the highest decalcification rates) where adult stages showed the lowest performance. On the other hand, A. purpuratus survivorship was not significantly affected by upwelling intensity which would be providing evidence of the high physiological flexibility and well-locally adapted is this species to fluctuating and occasional stressful environmental conditions. Our results are significantly relevant in the climate change context as some upwelling systems are at risk to change shortly (i.e., an upwelling intensification in frequency and intensity) as a consequence of changes in the atmospheric pressures that modulate favourable-upwelling winds. These changes may certainly increase the climate related-risks of the entire socio-ecological systems related to the fishery and aquaculture of A. purpuratus along the Humboldt Current System

Development of solvent-casting particulate leaching (SCPL) polymer scaffolds as improved three-dimensional supports to mimic the bone marrow niche

The need for new approaches to investigate ex vivo the causes and effects of tumor and to achieve improved cancer treatments and medical therapies is particularly urgent for malignant pathologies such as lymphomas and leukemias, whose tissue initiator cells interact with the stroma creating a three-dimensional (3D) protective environment that conventional mono- and bi-dimensional (2D) models are not able to simulate realistically. The solvent-casting particulate leaching (SCPL) technique, that is already a standard method to produce polymer-based scaffolds for bone tissue repair, is proposed here to fabricate innovative 3D porous structures to mimic the bone marrow niche in vitro. Two different polymers, namely a rigid polymethyl methacrylate (PMMA) and a flexible polyurethane (PU), were evaluated to the purpose, whereas NaCl, in the form of common salt table, resulted to be an efficient porogen. The adoption of an appropriate polymer-to-salt ratio, experimentally defined as 1:4 for both PMMA and PU, gave place to a rich and interconnected porosity, ranging between 82.1 vol% and 91.3 vol%, and the choice of admixing fine-grained or coarse-grained salt powders allowed to control the final pore size. The mechanical properties under compression load were affected both by the polymer matrix and by the scaffold's architecture, with values of the elastic modulus indicatively varying between 29 kPa and 1283 kPa. Preliminary tests performed with human stromal HS-5 cells co-cultured with leukemic cells allowed us to conclude that stromal cells grown associated to the supports keep their well-known protective and pro-survival effect on cancer cells, indicating that these devices can be very useful to mimic the bone marrow microenvironment and therefore to assess the efficacy of novel therapies in pre-clinical studies

Natural genetic variation for grapevine phenology as a tool for climate change adaptation

Grapevine phenology is being modified by climate change, particularly by the increase of temperatures that affect grape attributes for wine production. Besides the existing oenological and viticultural approaches, the thorough exploration of the current intra-cultivar genetic variability to select late-ripening genotypes emerges as an interesting alternative. In the present work, we have analyzed the natural genetic variation for phenology and agronomic traits among 21 'Malbec' clones and we demonstrated that fruiting cuttings are a useful tool for the analysis of such variation in 'Malbec'. Several clones could be distinguished by agronomic traits like berry number or cluster weight, and mainly by phenology characteristics like the length of the phase between flowering and veraison, which reached more than 16 days between early and late clones. These results support the approach of exploring grapevine clone collections in searching for genotypes with delayed phenology, and thus with the potential to maintain some expected quality characteristics under warm conditions.Fil: van Houten, Silvina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Biología Agrícola de Mendoza. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias. Instituto de Biología Agrícola de Mendoza; ArgentinaFil: Muñoz, Claudio Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Biología Agrícola de Mendoza. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias. Instituto de Biología Agrícola de Mendoza; ArgentinaFil: Bree, Laura. Vivero Mercier; ArgentinaFil: Bergamín, Daniel. Vivero Mercier; ArgentinaFil: Sola, Cristobal. Vivero Mercier; ArgentinaFil: Lijavetzky, Diego Claudio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Biología Agrícola de Mendoza. Universidad Nacional de Cuyo. Facultad de Ciencias Agrarias. Instituto de Biología Agrícola de Mendoza; Argentin

Algoritmo para el diagnóstico precoz de la deficiencia de vitamina B12 en ancianos

Background: The elderly population is particularly at risk for developing vitamin B12-deficiency. Serum cobalamin does not necessarily reflect a normal B12 status. The determination of methylmalonic acid is not available in all laboratories. Issues of sensitivity for holotranscobalamin and the low specificity of total homocysteine limit their utility. The aim of the present study is to establish a diagnostic algorithm by using a combination of these markers in place of a single measurement. Methods: We compared the diagnostic efficiency of these markers for detection of vitamin B12 deficiency in a population (n = 218) of institutionalized elderly (median age 80 years). Biochemical, haematological and morphological data were used to categorize people with or without vitamin B12 deficiency. Results: In receiver operating curves characteristics for detection on vitamin B12 deficiency using single measurements, serum folate has the greatest area under the curve (0.87) and homocysteine the lowest (0.67). The best specificity was observed for erythrocyte folate and methylmalonic acid (100% for both) but their sensitivity was very low (17% and 53%, respectively). The highest sensitivity was observed for homocysteine (81%) and serum folate (74%). When we combined these markers, starting with serum and erythrocyte folate, followed by holotranscobalamin and ending by methylmalonic acid measurements, the overall sensitivity and specificity of the algorithm were 100% and 90%, respectively. Conclusion: The proposed algorithm, which combines erythrocyte folate, serum folate, holotranscobalamin and methylmalonic acid, but eliminate B12 and tHcy measurements, is a useful alternative for vitamin B12 deficiency screening in an elderly institutionalized cohort.Introducción: Los mayores son una población que presenta un riesgo importante de desarrollar una deficiencia de vitamina B12, pero las concentraciones de cobalamina en suero no reflejan necesariamente un estado abnormal en el estado de B12 . Existen biomarcadores asociados a la vitamina B12: el ácido metilmalónico no está disponible en todos los laboratorios, la holotranscobalamina es poco sensible y la homocisteína presenta una baja especificidad. El objetivo del presente estudio es establecer un algoritmo de diagnóstico mediante el uso de una combinación de estos biomarcadores en lugar de la medición de uno sólo de ellos. Métodos: Se comparó la eficacia diagnóstica de estos marcadores para la detección de deficiencia de vitamina B12 en una población (n = 218) de ancianos institucionalizados (edad media 80 años). Los parámetros bioquímicos, hematológicos y morfológicos fueron utilizados para clasificar a los sujetos con o sin deficiencia de vitamina B12. Resultados: Se establecieron las curvas ROC (Receiver Operating Curves) para determinar la eficacia diagnóstica de cada parámetro, tomado individualmente. El folato sérico tenía la mayor área bajo la curva (0,87) y la homocisteína la más baja (0,67). Se observó que la mejor especificidad la presentaba el folato eritrocitario y el ácido metilmalónico (100% para ambos), pero sus sensibilidades eran muy bajas (17% y 53%, respectivamente). Y se observó que la sensibilidad más alta la presentaba la homocisteína (81%) y el folato sérico (74%), pero en contrapartida una especificidad baja. Cuando se combinaron estos marcadores, iniciando las determinaciones con el folato sérco y eritrocitario, seguido por holotranscobalamina y terminando por las mediciones de ácido metilmalónico, la sensibilidad y especificidad global del algoritmo fueron 100% y 90%, respectivamente. Conclusión: El algoritmo propuesto, que combina la determinación de folato sérico y eritrocitario, holotranscobalamina y ácido metilmalónico, sin necesidad de evaluar la vitamina B12 y la homocisteína, es una alternativa útil para la detección de un estado abnormal del estado de vitamina B12 en una población de ancianos institucionalizados

RECONSTRUCCIÓN DEL DIENTE ENDODONCIADO: A PROPÓSITO DE DOS CASOS

El tratamiento endodóntico es un recurso indiscutible para la rehabilitación dental, ofreciendo la posibilidad de conservar el diente con una gran tasa de éxito. Como todo tratamiento odontológico, este presenta una serie de complicaciones en el diente natural debido principalmente al aumento de su fragilidad por consecuencia de una gran pérdida de su estructura dental. La restauración post endodóntica constituye un gran reto clínico odontológico. Esta requiere un amplio y profundo conocimiento, resaltando factores como: grado de destrucción, valoración del estado periodontal, situación en la arcada, material de reconstrucción, etc. El objetivo de este trabajo es conocer las distintas opciones de restauración para el diente endodonciado, exponiendo las diferentes posibilidades terapéuticas basadas en la evidencia científica, a partir de un diagnóstico integral e individualizado de dos pacientes que acuden al Servicio de Prácticas Odontológicas.<br /