A transcribed enhancer dictates mesendoderm specification in pluripotency.

Enhancers and long noncoding RNAs (lncRNAs) are key determinants of lineage specification during development. Here, we evaluate remodeling of the enhancer landscape and modulation of the lncRNA transcriptome during mesendoderm specification. We sort mesendodermal progenitors from differentiating embryonic stem cells (ESCs) according to Eomes expression, and find that enhancer usage is coordinated with mesendoderm-specific expression of key lineage-determining transcription factors. Many of these enhancers are associated with the expression of lncRNAs. Examination of ESC-specific enhancers interacting in three-dimensional space with mesendoderm-specifying transcription factor loci identifies MesEndoderm Transcriptional Enhancer Organizing Region (Meteor). Genetic and epigenetic manipulation of the Meteor enhancer reveal its indispensable role during mesendoderm specification and subsequent cardiogenic differentiation via transcription-independent and -dependent mechanisms. Interestingly, Meteor-deleted ESCs are epigenetically redirected towards neuroectodermal lineages. Loci, topologically associating a transcribed enhancer and its cognate protein coding gene, appear to represent therefore a class of genomic elements controlling developmental competence in pluripotency

Harmful Elements in Estuarine and Coastal Systems

Estuaries and coastal zones are dynamic transitional systems which provide many economic and ecological benefits to humans, but also are an ideal habitat for other organisms as well. These areas are becoming contaminated by various anthropogenic activities due to a quick economic growth and urbanization. This chapter explores the sources, chemical speciation, sediment accumulation and removal mechanisms of the harmful elements in estuarine and coastal seawaters. It also describes the effects of toxic elements on aquatic flora and fauna. Finally, the toxic element pollution of the Venice Lagoon, a transitional water body located in the northeastern part of Italy, is discussed as a case study, by presenting the procedures adopted to measure the extent of the pollution, the impacts on organisms and the restoration activities

Pulmonary exacerbation score in Cystic Fibrosis patients: Reliability and validity.

Background: Lung disease in cystic fibrosis (CF) is characterized by recurrent pulmonary exacerbations (PEs), but consensus on diagnostic criteria for PE is lacking. The use of a consistent definition of PE as an outcome measure in CF clinical trials would allow meaningful comparison across centers. The aim of this study was to assess the reliability and validity of a simplified version of the Seattle Pulmonary Exacerbation Score (SPEX). Materials and Methods: A cross-sectional observational study with review of case notes was conducted on pediatric patients with CF in an outpatient setting. Inter-investigator reliability was assessed using the kappa coefficient of agreement, and intra-investigator reliability was examined following re-evaluation 21 months after the initial assessment. The validity of the SPEX was analyzed using independent clinical assessment as the "gold standard." The performance of the original and simplified scores was compared. Results: Inter- and intra-investigator reliability of SPEX scores were excellent (κ = 0.91 and 0.98, respectively). Validity testing yielded a kappa coefficient of 0.63. The sensitivity and specificity of the SPEX in detecting PE were 89.4% and 84%, respectively. The SPEX performed as well as the original measure. Conclusions: The SPEX is objective and repeatable. This quick and simple-to-use measure performed as well as the original version and is applicable to a real-life pediatric population outside of the context of narrowly defined clinical parameters. The use of the SPEX to diagnose PE consistently in children with CF is thus recommended

Covalent immobilization of native biomolecules onto Au(111) via N-hydroxysuccinimide ester functionalized self-assembled monolayers for scanning probe microscopy.

We have worked out a procedure for covalent binding of native biomacromolecules on flat gold surfaces for scanning probe microscopy in aqueous buffer solutions and for other nanotechnological applications, such as the direct measurement of interaction forces between immobilized macromolecules, of their elastomechanical properties, etc. It is based on the covalent immobilization of amino group-containing biomolecules (e.g., proteins, phospholipids) onto atomically flat gold surfaces via omega-functionalized self-assembled monolayers. We present the synthesis of the parent compound, dithio-bis(succinimidylundecanoate) (DSU), and a detailed study of the chemical and physical properties of the monolayer it forms spontaneously on Au(111). Scanning tunneling microscopy and atomic force microscopy (AFM) revealed a monolayer arrangement with the well-known depressions that are known to stem from an etch process during the self-assembly. The total density of the omega-N-hydroxysuccinimidyl groups on atomically flat gold was 585 pmol/cm(2), as determined by chemisorption of (14)C-labeled DSU. This corresponded to approximately 75% of the maximum density of the omega-unsubstituted alkanethiol. Measurements of the kinetics of monolayer formation showed a very fast initial phase, with total coverage within 30 S. A subsequent slower rearrangement of the chemisorbed molecules, as indicated by AFM, led to a decrease in the number of monolayer depressions in approximately 60 min. The rate of hydrolysis of the omega-N-hydroxysuccinimide groups at the monolayer/water interface was found to be very slow, even at moderately alkaline pH values. Furthermore, the binding of low-molecular-weight amines and of a model protein was investigated in detail

A review of virtual planning software for guided implant surgery - Data import and visualization, drill guide design and manufacturing

Background: Virtual implant planning systems integrate (cone beam-) computed tomography data to assess bone quantity and virtual models for the design of the implant-retained prosthesis and drill guides. Five commercially available systems for virtual implant planning were examined regarding the modalities of integration of radiographic data, virtual dental models and the design of drill guides for guided implant surgery. The purpose of this review was to describe the limitations of these available systems regarding the import of imaging data and the design and fabrication of a drill guide. Methods: The following software systems were examined regarding the import of imaging data and the export of the virtual implant planning for the design and fabrication of a drill guide with the help of two clinical situations requiring dental implant therapy: coDiagnostiX™, DentalWings, Canada (CDX); Simplant Pro™, Dentsply, Sweden (SIM); Smop™, Swissmeda, Switzerland (SMP); NobelClinician™, Nobel Biocare, Switzerland (NC); Implant Studio, 3Shape, Denmark (IST). Assessment criteria included data formats and management as well as the workflow for the design and production of drill guides. Results: All systems have a DICOM-interface ("Digital Imaging and Communication in Medicine") for the import of radiographic data. Imaging artefacts could be reduced but not eliminated by manual data processing. The import of virtual dental models in a universal format (STL: Standard Tesselation Language) was possible with three systems; one system could only be used with a proprietary data format. All systems display three-dimensional surface models or two-dimensional cross-sections with varying orientation for virtual implant planning. Computer aided design and manufacturing (CAD/CAM) of drill guides may be performed by the user with the help of default parameters or solely by the provider of the software and thus without the influence of the clinician. Conclusion: Data bases of commonly used implant systems are available in all tested software, however not all systems allow to plan and execute fully guided implant placement. An individual design and in-house manufacturing of the drill guide is only available in some software systems. However, at the time of publication most recent software versions showed flexibility in individual design and in-house manufacturing of drill guides