The GABAA Receptor delta Subunit Gene Promoter: Characterisation and Use

Viral vectors represent powerful tools in the rapidly developing field of gene therapy of the mammalian central nervous system. Interest in HSV-1 in particular as a vector relates primarily to its ability to establish long-term latent infections in neuronal cells. A goal of this research was the development of a safe HSV-1 vector that would deliver a reporter gene to target cells in the CNS and restrict its expression to a specific cell-type by the use of a neuronal promoter. A second goal was to add to the present limited understanding of neuron-specific promoter structure. The murine GABAA receptor delta subunit gene promoter was selected for use as a prototype neuronal promoter as it is moderately expressed and has a well characterised restricted expression pattern in the CNS. Characterisation of the 5'- romoter region of the GABAA receptor 6 subunit gene was achieved by sequencing 10.5 kb of DNA from the 5'-upstream region. Analysis of this sequence revealed the presence of several putative recognition sites for transcription factors. Identification of the transcription start points showed two main clusters of start sites located 58 and 108 bases upstream from the translational start point. Comparison of about 3.5 kb of DNA sequence of the promoters of the rat and mouse genes showed a high degree of conservation of noncoding sequence between the species. The recognition site for a putative regulatory factor BSF 1 identified in the 5'-upstream sequence of the rat delta gene was demonstrated to be absent in the mouse 5'-flanking sequence. Analysis of a range of promoter - reporter constructs in the NB4 1A3 cell line suggested the presence of a silencing element located between 4.5 kb and 6.3 kb upstream of the translational start point. As a preliminary step to evaluating in vivo mutagenesis investigations of GABAA receptor 5 subunit gene promoter function, targeted disruption of the GABAA receptor 6 subunit gene in embryonic stem cells was achieved. Two genomic DNA fragments from the GABAA receptor 5 subunit gene were selected and inserted into a simple replacement vector (pNT) containing the neomycin gene and the HSV thymidine kinase gene. A successful targeting event would result in the removal of the first exon, 4.5 kb of promoter sequence and 2 kb of DNA from the first intron. Two targeted embryonic stem cell lines were isolated and can be used to make mice homozygous for the mutated gene. The GABAA receptor delta subunit gene had previously been assigned to human Chromosome Ip. Genetic mapping of the gene to rat chromosome 5 and mouse chromosome 4 was performed. These results agree with expected regions of synteny between human, mouse and rat. A range of recombinant HSV-1 viruses were produced which contained the E coli lacZ gene driven by different promoters. Two loci within the HSV-1 genome were chosen as sites of insertion, LAT and UL43 in two HSV-1 viral variants, 1716 and 1764. Infection of primary cerebellar granule cell cultures with a range of these recombinant viruses revealed that the cerebellar granule cells were not readily infected by this viral variant. Stereotactic injection of the viruses into the cerebellum of adult rats failed to show specific P-galactosidase expression. Nevertheless, the GABAA receptor delta subunit gene promoter-lacZ fusion constructs can now be transferred to new HSV-1 vectors with better growth characteristics thus furthering the original goal of this work

Expected Performance of the ATLAS Experiment - Detector, Trigger and Physics

A detailed study is presented of the expected performance of the ATLAS detector. The reconstruction of tracks, leptons, photons, missing energy and jets is investigated, together with the performance of b-tagging and the trigger. The physics potential for a variety of interesting physics processes, within the Standard Model and beyond, is examined. The study comprises a series of notes based on simulations of the detector and physics processes, with particular emphasis given to the data expected from the first years of operation of the LHC at CERN

The development and validation of a scoring tool to predict the operative duration of elective laparoscopic cholecystectomy

Background: The ability to accurately predict operative duration has the potential to optimise theatre efficiency and utilisation, thus reducing costs and increasing staff and patient satisfaction. With laparoscopic cholecystectomy being one of the most commonly performed procedures worldwide, a tool to predict operative duration could be extremely beneficial to healthcare organisations. Methods: Data collected from the CholeS study on patients undergoing cholecystectomy in UK and Irish hospitals between 04/2014 and 05/2014 were used to study operative duration. A multivariable binary logistic regression model was produced in order to identify significant independent predictors of long (> 90 min) operations. The resulting model was converted to a risk score, which was subsequently validated on second cohort of patients using ROC curves. Results: After exclusions, data were available for 7227 patients in the derivation (CholeS) cohort. The median operative duration was 60 min (interquartile range 45–85), with 17.7% of operations lasting longer than 90 min. Ten factors were found to be significant independent predictors of operative durations > 90 min, including ASA, age, previous surgical admissions, BMI, gallbladder wall thickness and CBD diameter. A risk score was then produced from these factors, and applied to a cohort of 2405 patients from a tertiary centre for external validation. This returned an area under the ROC curve of 0.708 (SE = 0.013, p  90 min increasing more than eightfold from 5.1 to 41.8% in the extremes of the score. Conclusion: The scoring tool produced in this study was found to be significantly predictive of long operative durations on validation in an external cohort. As such, the tool may have the potential to enable organisations to better organise theatre lists and deliver greater efficiencies in care

Drons col·laboratius

La robòtica col·laborativa és senzillament robots dissenyats per dur a terme treballs de col·laboració amb els humans. Els robots col·laboratius o cobots són cada cop més utilitzats a les indústries. La robòtica col·laborativa és un dels àmbits d'actualitat en aquests moments. Però també és un dels més interessants en més d'un sentit. Com es comuniquen dos drons autònoms que col·laboren per fer una tasca? Com són aquests missatges que s'envien? Que poden fer que no podrien fer sols? Aquestes són algunes de les preguntes que ens volem respondre en aquest projecte. En aquest treball es presenta un disseny i implementació de dos drons terrestres que es comuniquen per col·laborar entre ells per resoldre una tasca.Collaborative robotics is simply robots designed to perform collaborative work with humans. Collaborative robots or cobots are increasingly used in industries. Collaborative robotics is one of the current topics now. But it is also one of the most interesting in more ways than one. How do two autonomous drones that collaborate to perform a task communicate? How are these messages sent? What can they do that they could not do alone? These are some of the questions we want to answer in this project. This work presents a design and implementation of two ground drones that communicate to collaborate with each other to solve a task.La robótica colaborativa es sencillamente robots diseñados para llevar a cabo trabajos de colaboración con los humanos. Los robots colaborativos o cobots son cada vez más utilizados en las industrias. La robótica colaborativa es uno de los ámbitos de actualidad. Pero también es uno de los más interesantes en más de un sentido. ¿Cómo se comunican drones autónomos que colaboran para hacer una tarea? ¿Cómo son estos mensajes que es envían? ¿Qué pueden hacer que no lo podrían hacer solos? Estas son algunas de las preguntas que queremos responder con este proyecto. En este trabajo se presenta un diseño e implementación de dos drones terrestres que se comunican para colaborar entre ellos para resolver una tarea

Archaeological assessment reveals Earth's early transformation through land use

Environmentally transformative human use of land accelerated with the emergence of agriculture, but the extent, trajectory, and implications of these early changes are not well understood. An empirical global assessment of land use from 10,000 years before the present (yr B.P.) to 1850 CE reveals a planet largely transformed by hunter-gatherers, farmers, and pastoralists by 3000 years ago, considerably earlier than the dates in the land-use reconstructions commonly used by Earth scientists. Synthesis of knowledge contributed by more than 250 archaeologists highlighted gaps in archaeological expertise and data quality, which peaked for 2000 yr B.P. and in traditionally studied and wealthier regions. Archaeological reconstruction of global land-use history illuminates the deep roots of Earth's transformation and challenges the emerging Anthropocene paradigm that large-scale anthropogenic global environmental change is mostly a recent phenomenon

Archaeological assessment reveals Earth’s early transformation through land use

Humans began to leave lasting impacts on Earth's surface starting 10,000 to 8000 years ago. Through a synthetic collaboration with archaeologists around the globe, Stephens et al. compiled a comprehensive picture of the trajectory of human land use worldwide during the Holocene (see the Perspective by Roberts). Hunter-gatherers, farmers, and pastoralists transformed the face of Earth earlier and to a greater extent than has been widely appreciated, a transformation that was essentially global by 3000 years before the present.Science, this issue p. 897; see also p. 865Environmentally transformative human use of land accelerated with the emergence of agriculture, but the extent, trajectory, and implications of these early changes are not well understood. An empirical global assessment of land use from 10,000 years before the present (yr B.P.) to 1850 CE reveals a planet largely transformed by hunter-gatherers, farmers, and pastoralists by 3000 years ago, considerably earlier than the dates in the land-use reconstructions commonly used by Earth scientists. Synthesis of knowledge contributed by more than 250 archaeologists highlighted gaps in archaeological expertise and data quality, which peaked for 2000 yr B.P. and in traditionally studied and wealthier regions. Archaeological reconstruction of global land-use history illuminates the deep roots of Earth's transformation and challenges the emerging Anthropocene paradigm that large-scale anthropogenic global environmental change is mostly a recent phenomenon

IRBs and Best Practices for Ethical Data Sharing

Materials associated with 2017-07-30 sessio