Eukaryotic protein-coding genes are transcribed by RNA polymerase II (RNAPII) through a cycle composed of three main phases: initiation, elongation, and termination. Recent studies using chromatin immunoprecipitation coupled to high-throughput sequencing suggest that the density of RNAPII molecules is higher at the 3'-end relative to the gene body. Here we show that this view is biased due to averaging density profiles for "metagene" analysis. Indeed, the majority of genes exhibit little, if any, detectable accumulation of polymerases during transcription termination. Compared with genes with no enrichment, genes that accumulate RNAPII at the 3'-end are shorter, more frequently contain the canonical polyadenylation [poly(A)] signal AATAAA and G-rich motifs in the downstream sequence element, and have higher levels of expression. In 1% to 4% of actively transcribing genes, the RNAPII enriched at the 3'-end is phosphorylated on Ser5, and we provide evidence suggesting that these genes have their promoter and terminator regions juxtaposed. We also found a striking correlation between RNAPII accumulation and nucleosome organization, suggesting that the presence of nucleosomes after the poly(A) site induces pausing of polymerases, leading to their accumulation. Yet we further observe that nucleosome occupancy at the 3'-end of genes is dynamic and correlates with RNAPII density. Taken together, our results provide novel insight to transcription termination, a fundamental process that remains one of the least understood stages of the transcription cycle.

From a user centered point of view an important basic requirement to enable human-robot cooperation is to achieve conformity with operator's expectations of robot behavior. Therefore, this study focuses on the question, whether anthropomorphic robot movement trajectories can lead to an improved anticipation of the robot's behavior. Based on a virtual simulation environment a robotized assembly cell consisting of the assembly robot and the actual workplace was considered. In order to be able to simulate anthropomorphic movements, the human wrist trajectories of defined pick and place movements were obtained using an infrared motion capture system. The captured data were used to navigate the virtual assembly robot. Within the experiment anthropomorphic and robotic trajectories were distinguished. During the experiment, the main task of the participants was to predict the movement's destination as quickly as possible. Thus, the corresponding reaction value was analyzed to investigate the influence of anthropomorphic robot movements on human prediction in industrial environments.

Implementations of Software Transactional Memory (STM) algorithms associate metadata with the memory locations accessed during a transaction’s lifetime. This metadata may be stored either in-place, by wrapping every memory cell in a container that includes the memory cell itself and the corresponding metadata; or out-place (also called external), by resorting to a mapping function that associates the memory cell address with an external table entry containing the corresponding metadata. The implementation techniques for these two approaches are very different and each STM framework is usually biased towards one of them, only allowing the efficient implementation of STM algorithms following that approach, hence inhibiting the fair comparison with STM algorithms falling into the other. In this paper we introduce a technique to implement in-place metadata that does not wrap memory cells, thus overcoming the bias by allowing STM algorithms to directly access the transactional metadata. The proposed technique is available as an extension to the DeuceSTM framework, and enables the efficient implementation of a wide range of STM algorithms and their fair (unbiased) comparison in a common STM infrastructure. We illustrate the benefits of our approach by analyzing its impact in two popular TM algorithms with two different transactional workloads, TL2 and multi-versioning, with bias to out-place and in-place respectively.

In this work, we propose RATS, a middleware to enhance and extend the Terracotta framework for Java with the ability to transparently execute multi-threaded Java applications to provide a single-system image. It supports efficient scheduling of threads, according to available resources, across several nodes in a Terracotta cluster, taking advantage of the extra computational and memory resources available. It also supports profiling to gather application characteristics such as dispersion of thread workload, thread inter-arrival time and resource usage of the application. Profiling and clustering capabilities are inserted with the help of byte code instrumentations. We developed a range of alternative scheduling heuristics and classify them based on the application and cluster behavior. The middleware is tested with different applications with varying thread characteristics to assess and classify the scheduling heuristics with respect to application speed-ups. Results indicate that, for a CPU intensive application, it is possible to classify the scheduling heuristic based on application and cluster properties and also achieve linear speed ups. Furthermore, we show that a memory intensive application is able to scale its memory usage considerably when compared to running the application on a single JVM.

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Ion Jelly materials combine the chemical versatility and conductivity of an ionic liquid (IL) with the morphological versatility of a biopolymer (gelatin). They exhibit very interesting properties, such as conductivities up to 10− 4 S cm− 1, and high thermostability up to 180 °C, and have been used successfully to design electrochromic windows. In this work we report on the preparation of Ion Jelly fibers through electrospinning in order to obtain high surface area conductive materials. We have used the IL 1-(2-hydroxyethyl)-3-methyl-imidazolium tetrafluoroborate ([C2OHmim]BF4), which exhibits conveniently high ionic conductivity (over 10− 3 S cm− 1) and electrochemical stability (electrochemical window over 6.0 V). The morphology of the obtained fibers was quantified using Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM). We found that on average the effect of the IL on fiber diameter differs for lower and higher IL concentrations and that this effect was correlated with the initial conductivity and viscosity of Ion Jelly electrospinning solution. Moreover we also found that conductivities of Ion Jelly fibers are of the same order of magnitude as the conductivities of Ion Jelly dense films (~ 10− 4 S cm− 1). To the best of our knowledge, this is the first report on the incorporation of an IL into gelatin fibers using electrospinning. This opens up new opportunities for the application of gelatin fibers in electrochemical and biomedical devices.

This work reports the production of hydroxyapatite (HA) sub-micron fibers by combining electrospinning and a non-alkoxide sol–gel system, using cheap precursors. Phosphorus pentoxide (P2O5) and calcium nitrate tetrahydrate (Ca(NO3)2.4H2O) were used as precursors of phosphorus and calcium, respectively. The fibers were electrospun from a mixture of the gel formed from the system Ca(NO3)2.4H2O/P2O5 with polymeric solutions of polyvinylpyrrolidone (PVP) in water and ethanol/water mixtures. The fibers were analyzed for their morphology (Scanning Electron Microscopy, SEM), chemical composition (Fourier Transform Infrared Spectroscopy, FTIR) and structure (X-ray diffraction, XRD). The fibers obtained were composed mainly of type B carbonated HA with traces of β-tricalcium phosphate (β-TCP). SEM analysis revealed that increasing the concentration of water in the solvent system, used in the preparation of electrospinning solutions, led to fibers with smaller diameters and narrower diameter distribution.

The bacterium Geobacter sulfurreducens (Gs) can grow in the presence of extracellular terminal acceptors, a property that is currently explored to harvest electricity from aquatic sediments and waste organic matter into microbial fuel cells. A family composed of five triheme cytochromes (PpcA-E) was identified in Gs. These cytochromes play a crucial role by bridging the electron transfer from oxidation of cytoplasmic donors to the cell exterior and assisting the reduction of extracellular terminal acceptors. The detailed thermodynamic characterization of such proteins showed that PpcA and PpcD have an important redox-Bohr effect that might implicate these proteins in the e−/H+ coupling mechanisms to sustain cellular growth. The physiological relevance of the redox-Bohr effect in these proteins was studied by determining the fractional contribution of each individual redox-microstate at different pH values. For both proteins, oxidation progresses from a particular protonated microstate to a particular deprotonated one, over specific pH ranges. The preferred e−/H+ transfer pathway established by the selected microstates indicates that both proteins are functionally designed to couple e−/H+ transfer at the physiological pH range for cellular growth.

In this work we have evaluated the potential of boronic acid functionalized magnetic particles for the one-step capture of a human monoclonal antibody (mAb) from a Chinese hamster ovary (CHO) cell culture supernatant. For comparison, Protein A coated magnetic particles were also used. The most important factor influencing the overall process yield and product purity in boronic acid particles was found to be the binding pH. Basic pH values promoted higher purities while resulting in decreased yields due to the competing effects of molecules such as glucose and lactate present in the cell culture supernatant. After optimization, the particles were successfully used in a multi-cycle purification process of the mAb from the CHO feedstock. Boronic acid particles were able to achieve an average overall yield of 86% with 88% removal of CHO host cell proteins (HCP) when the binding was performed at pH 7.4, while at pH 8.5 these values were 58% and 97%, respectively. In both cases, genomic DNA removal was in excess of 97%. Comparatively, Protein A particles recorded an average overall yield of 80% and an HCP removal greater than 99%. The adsorption of the mAb to the boronic acid particles was shown to be mediated by strong affinity interactions. Overall, boronic acid based purification processes can offer a cost-effective alternative to Protein A as the direct capturing step from the mammalian cell culture.

Fluorescence microscopy and microspectrofluorometry are important tools in the characterization and identification of proteins, offering a great range of applications in conservation science. Because of their high selectivity and sensitivity, the combination of these techniques can be exploited for improved recognition and quantification of proteinaceous binders in paintings and polychromed works of art. The present article explores an analytical protocol integrating fluorescence microscopy and fluorometry for both identification and mapping of proteinaceous binders (in particular egg and glues) in paint samples. The study has been carried out on historically accurate reconstructions simulating the structure and composition of tempera and oil paints containing these binders. To assess the spatial distribution of specific proteins within the paint layers, cross-sections from the reconstructions were analyzed by fluorescence imaging after staining with an exogenous fluorophore. Reference fluorescence spectra for each layer were acquired by a multichannel spectral analyzer and compared after Gaussian deconvolution. The results obtained demonstrated the effectiveness of the integrated protocol, highlighting the potential for the use of fluorescent staining coupled with microspectrofluorometry as a routine diagnostic tool in conservation science. The current work creates a set of fully characterized reference samples for further comparison with those from actual works of art.

Several Software Transactional Memory (STM) algorithms have been developed in the last few years. These algorithms differ in many different properties, such as progress and safety guarantees, contention management, etc. Some STM frameworks (DSTM2, DeuceSTM) were developed to allow the implementation and comparison of many different algorithms using the same transactional interface. The information required by an STM algorithm is either associated with the execution of a transaction (frequently referred as transaction descriptor), or associated with each memory location (or object reference) accessed within the transaction. The transaction descriptor is typically stored in a thread-local memory space and maintains the information necessary to validate and commit a memory transaction. The information associated with each memory location depends on the nature of the STM algorithm, which we will henceforth refer to as metadata, and may be composed by e.g. locks, timestamps or version lists. Since metadata is associated with each memory location, there are two main strategies regarding the location for storing such metadata: The metadata is stored either near each memory location (in-place strategy); or in a mapping table that associates the metadata with the corresponding memory location (out- place strategy). DeuceSTM is one of the most efficient STM frameworks available for the Java programming language and provides a well defined interface that allows to implement several STM algorithms using an out-place strategy. This work describes the adaptation and extension of DeuceSTM to support the in-place metadata strategy. Our approach complies to the following properties: Efficiency –- The STM implementation does not rely on an auxiliary mapping table, thus providing faster direct access to the transactional metadata; Non-transactional code is oblivious to the presence of metadata in objects, hence there is no performance overhead whatsoever for non-transactional code; Transactional code avoids the extra memory dereference imposed by the decorator pattern; Primitive types are fully supported, even in transactional code; And we provide a solution for fully supporting transactional N-dimensional arrays, which again impose no overhead on non-transactional code. Transparency –- The STM implementation automatically identifies, creates and initializes the necessary additional metadata fields in the objects; Non-transactional code is oblivious to the presence of metadata in objects, hence no code changes are required; The new transactional array types (supporting metadata for individual cells) are compatible with the standard arrays, hence not requiring any pre-/post- processing of the arrays when invoking standard or third-party non-transactional libraries. Performance benchmarking has confirmed that our approach for supporting in-place metadata in DeuceSTM, alongside with the existing out-place strategy, is a valid functional complement, widening the range of algorithms that can be efficiently implemented in DeuceSTM, enabling their fair comparison in a common infrastructure.