Surface guidance of stem cell behavior: Chemically tailored co-presentation of integrin-binding peptides stimulates osteogenic differentiation in vitro and bone formation in vivo

Surface modification stands out as a versatile technique to create instructive biomaterials that are able to actively direct stem cell fate. Chemical functionalization of titanium has been used in this work to stimulate the differentiation of human mesenchymal stem cells (hMSCs) into the osteoblastic lineage, by covalently anchoring a synthetic double-branched molecule (PTF) to the metal that allows a finely controlled presentation of peptidic motifs. In detail, the effect of the RGD adhesive peptide and its synergy motif PHSRN is studied, comparing a random distribution of the two peptides with the chemically-tailored disposition within the custom made synthetic platform, which mimics the interspacing between the motifs observed in fibronectin. Contact angle measurement and XPS analysis are used to prove the efficiency of functionalization. We demonstrate that, by rationally designing ligands, stem cell response can be efficiently guided towards the osteogenic phenotype: In vitro, PTF-functionalized surfaces support hMSCs adhesion, with higher cell area and formation of focal contacts, expression of the integrin receptor a5ß1 and the osteogenic marker Runx2, and deposition a highly mineralized matrix, reaching values of mineralization comparable to fibronectin. Our strategy is also demonstrated to be efficient in promoting new bone growth in vivo in a rat calvarial defect. These results highlight the efficacy of chemical control over the presentation of bioactive peptides; such systems may be used to engineer bioactive surfaces with improved osseointegrative properties, or can be easily tuned to generate multi-functional coatings requiring a tailored disposition of the peptidic motifs.Peer ReviewedPostprint (author's final draft

Sensory and physical changes in chilled farmed coho salmon (Oncorhynchus kisutch): Effect of previous optimized hydrostatic high-pressure conditions

11 páginas, 2 figuras, 4 tablas.-- The final publication is available at www.springerlink.comThe effect of a previous hydrostatic high-pressure (HHP) treatment on sensory and physical quality of chilled coho salmon (Oncorhynchus kisutch) was investigated. As a first stage, a 22 factorial design based on the response surface methodology was used for optimization of HHP conditions; for it, the effects on color uniformity, white spots appearance, elasticity, and hardness were analyzed. In a second stage, optimized HHP conditions (135 MPa for 30 s, 170 MPa for 30 s, and 200 MPa for 30 s; treatments T-1, T-2, and T-3, respectively) were applied as previous treatment to chilling, being sampling carried out on salmon muscle at days 0, 6, 10, 15, and 20. A marked effect on sensory and physical parameters was detected after HHP treatment and throughout the chilled storage. According to odor (rancid, putrid), texture (elasticity, gaping, firmness), and color (L* value) attributes, fish corresponding to T-1 condition was shown to have better-maintained quality throughout the chilled storage than fish belonging to T-2 and T-3 conditions; additionally, a quality enhancement (putrid odor, cohesivity, gaping) was found when compared with control samples. In agreement to the preliminary optimization study, it is concluded that T-1 condition can provide the most effective HHP pre-treatment to better maintain sensory and physical properties when salmon muscle is to be commercialized as a fresh product. Results obtained for the quality evolution of chilled fish attest the importance of establishing a judicious selection of previous HHP treatment parameters to minimize undesirable changes related to sensory and physical characteristicsThis work was supported by the Universidad de Chile (Chile)-Consejo Superior de Investigaciones Científicas (Spain) program (Project 2006 CL 0034) and the FONDECYT program (Chile; project number: 1080626)Peer reviewe

Efecto de un tratamiento previo por altas presiones hidrostáticas sobre la alteración lipídica de jurel chileno (Trachurus murphyi) refrigerado

Lipid damage evolution was analyzed in chilled Chilean jack mackerel (Trachurus murphyi) previously treated with high hydrostatic pressure (HHP) technology. Different pressure levels and pressure holding times were tested. In addition, fish corresponding to pre- and post-rigor mortis (RM) stages were comparatively studied. Previous HHP treatment led to a marked lipid hydrolysis inhibition in chilled fish. Increasing the pressure level and pressure holding time led to a lower free fatty acid content, with the effect of pressure being more relevant. According to the analysis of different types of lipid oxidation indexes, no effect of the previous HHP treatment on the lipid oxidation development could be determined in chilled jack mackerel. Concerning the effect of the RM stage of raw fish, a higher primary and secondary lipid oxidation development was observed in fish corresponding to the post-RM condition throughout the chilled storage; although a definite effect on lipid hydrolysis could not be found.Se estudió la evolución de la alteración lipídica en jurel chileno (Trachurus murphyi) refrigerado previamente tratado a altas presiones hidrostáticas (HHP). Se aplicaron distintos valores de presión y tiempo de presurización; asimismo, se analizó de forma comparativa la respuesta al proceso del pescado inicial en estados pre- y post-rigor mortis (RM). El tratamiento previo por HHP produjo inhibición de la hidrólisis lipídica en pescado refrigerado, siendo más intenso el efecto de la presión que el del tiempo de presurización. De acuerdo con el análisis de distintos índices de oxidación, no se concluyó un efecto determinante sobre la oxidación lipídica por parte del tratamiento previo de HHP. En relación al efecto del estado de RM del pescado inicial, se observó una oxidación primaria y secundaria mayor en jurel correspondiente a la condición post-RM durante la conservación en refrigeración; sin embargo, no se detectó un efecto claro sobre la hidrólisis lipídica

Study on utilization status of internet and needs assessment for developing nutrition education programs among elementary school children

This study was to investigate utilization status of internet, health/nutrition websites among children, and to assess the needs for developing nutrition websites and education programs for children. The survey questionnaire was administered to 5-6th grade students (n=434) at two elementary schools. About 32% used the internet every day while 19.5% used it whenever they needed, showing significant differences in internet usage by gender (p<0.01). Although the subjects used the internet frequently, those who used health/nutrition websites were 23.3%. The purpose of using these sites were mainly 'to obtain health/nutrition information' (55%), 'to get information regarding weight control' (17%). Fifty-six percent of the users were satisfied with the nutrition websites, but only 30% said that they were helpful. The preferred topics in developing nutrition websites were assessment of obesity, exercise methods, weight control methods, nutrition information (e.g., diet for stature growth), dietary assessment and food hygiene. Girls showed more interest in these topics than boys (p<0.05). For school nutrition education, girls showed more interest than boys in topics for cooking snacks (p<0.001) and selecting snacks (p<0.05). In nutrition websites, subjects wanted to have information and game/quiz, as well as getting information using Flash animation. The favorite colors for screen and text were slightly different by gender (p<0.01). In school nutrition education, 89.5% of subjects liked to have activities (e.g., cooking, exercise, game). They also liked materials using computers, video and internet than printed materials. If nutrition education was done at schools, subjects wanted to receive 5.7 times of education per semester on average (mean length: 42.6 min./session). This study suggests that nutrition websites and education programs for children should include the topics such as assessment of obesity or diet, weight control and special information (e.g., diet for growth) as well as general information. In designing nutrition websites and programs, methods including game, quiz, Flash animation and activities (cooking, exercise) could be appropriately used to induce the interest and involvement of children

Multidrug resistant pulmonary tuberculosis treatment regimens and patient outcomes: an individual patient data meta-analysis of 9,153 patients.

Treatment of multidrug resistant tuberculosis (MDR-TB) is lengthy, toxic, expensive, and has generally poor outcomes. We undertook an individual patient data meta-analysis to assess the impact on outcomes of the type, number, and duration of drugs used to treat MDR-TB

Immunohistochemical identification and quantitative analysis of cytoplasmic Cu/Zn superoxide dismutase in mouse organogenesis

Cytoplasmic Cu/Zn superoxide dismutase (SOD1) is an antioxidant enzyme that converts superoxide to hydrogen peroxide in cells. Its spatial distribution matches that of superoxide production, allowing it to protect cells from oxidative stress. SOD1 deficiencies result in embryonic lethality and a wide range of pathologies in mice, but little is known about normal SOD1 protein expression in developing embryos. In this study, the expression pattern of SOD1 was investigated in post-implantation mouse embryos and extraembryonic tissues, including placenta, using Western blotting and immunohistochemical analyses. SOD1 was detected in embryos and extraembryonic tissues from embryonic day (ED) 8.5 to 18.5. The signal in embryos was observed at the lowest level on ED 9.5-11.5, and the highest level on ED 17.5-18.5, while levels remained constant in the surrounding extraembryonic tissues during all developmental stages examined. Immunohistochemical analysis of SOD1 expression on ED 13.5-18.5 revealed its ubiquitous distribution throughout developing organs. In particular, high levels of SOD1 expression were observed in the ependymal epithelium of the choroid plexus, ganglia, sensory cells of the olfactory and vestibulocochlear epithelia, blood cells and vessels, hepatocytes and hematopoietic cells of the liver, lymph nodes, osteogenic tissues, and skin. Thus, SOD1 is highly expressed at late stages of embryonic development in a cell- and tissue-specific manner, and can function as an important antioxidant enzyme during organogenesis in mouse embryos

Preparation and characterization of novel chitosan-based mixed matrix membranes resistant in alkaline media

In this work, mixed matrix membranes (MMMs) based on chitosan (CS) and different fillers (room temperature ionic liquid [emim][OAc] (IL), metallic Sn powder, layered titanosilicate AM-4 and layered stannosilicate UZAR-S3) were prepared by solution casting. The room temperature electrical conductivity and electrochemical response in strong alkaline medium were measured by electrochemical impedance spectroscopy and cyclic voltammetry (CV). The ionic conductivity of pure CS membranes was enhanced, from 0.070 to 0.126 mS cm21, for the pristine CS and Sn/CS membranes, respectively, as a function of the hydrophilic nature of the membrane and the coordination state of Sn. This hydrophilic and charge nature was corroborated by water uptake measurements, where only the introduction of IL in the CS membrane led to a water uptake of 3.96 wt %, 20 or 30 times lower than the other membranes. Good thermal and chemical stability in alkaline media were observed by thermogravimetric analyses and X-ray photoelectron spectroscopy analyses, respectively, and good interaction between CS and the fillers observed by X-ray diffraction, scanning electron microscopy and CV. Thus, thin CS-based MMMs (40–139 mm), resistant in high alkaline media, show higher conductivity than pure CS membranes, especially those fillers containing tin, and although the electrochemical performance is lower than commercially available anion-exchange membranes they have potential in pervaporation.This work has been funded by the Spanish MINECO through grants CTQ2010-20347, at the University of Alicante, and CTQ2012-31229 and RYC2011-08550, at the University of Cantabria. The authors gratefully thank Prof. Frank Marken, from the University of Bath (UK), for his advice on the electrochemical impedance characterization, and Dr. César Rubio, Dr. Carlos Téllez, and Prof. Joaquín Coronas, from the University of Zaragoza and the Instituto de Nanociencia de Aragón, Spain, for the UZAR-S3 sample and fruitful discussions

Fused Smart Sensor Network for Multi-Axis Forward Kinematics Estimation in Industrial Robots

Flexible manipulator robots have a wide industrial application. Robot performance requires sensing its position and orientation adequately, known as forward kinematics. Commercially available, motion controllers use high-resolution optical encoders to sense the position of each joint which cannot detect some mechanical deformations that decrease the accuracy of the robot position and orientation. To overcome those problems, several sensor fusion methods have been proposed but at expenses of high-computational load, which avoids the online measurement of the joint’s angular position and the online forward kinematics estimation. The contribution of this work is to propose a fused smart sensor network to estimate the forward kinematics of an industrial robot. The developed smart processor uses Kalman filters to filter and to fuse the information of the sensor network. Two primary sensors are used: an optical encoder, and a 3-axis accelerometer. In order to obtain the position and orientation of each joint online a field-programmable gate array (FPGA) is used in the hardware implementation taking advantage of the parallel computation capabilities and reconfigurability of this device. With the aim of evaluating the smart sensor network performance, three real-operation-oriented paths are executed and monitored in a 6-degree of freedom robot

Sequence-dependent DNA condensation as a driving force of DNA phase separation

The physical properties of DNA have been suggested to play a central role in spatio-temporal organization of eukaryotic chromosomes. Experimental correlations have been established between the local nucleotide content of DNA and the frequency of inter- and intra-chromosomal contacts but the underlying physical mechanism remains unknown. Here, we combine fluorescence resonance energy transfer (FRET) measurements, precipitation assays, and molecular dynamics simulations to characterize the effect of DNA nucleotide content, sequence, and methylation on inter-DNA association and its correlation with DNA looping. First, we show that the strength of DNA condensation mediated by poly-lysine peptides as a reduced model of histone tails depends on the DNA???s global nucleotide content but also on the local nucleotide sequence, which turns out to be qualitatively same as the condensation by spermine. Next, we show that the presence and spatial arrangement of C5 methyl groups determines the strength of inter-DNA attraction, partially explaining why RNA resists condensation. Interestingly, multi-color single molecule FRET measurements reveal strong anti-correlation between DNA looping and DNA-DNA association, suggesting that a common biophysical mechanism underlies them. We propose that the differential affinity between DNA regions of varying sequence pattern may drive the phase separation of chromatin into chromosomal subdomains