Miracle-dz/UniFAR-FTUs huggingface dataset:
Recovery of infectious respiratory syncytial virus expressing an additional, foreign gene

A previous report described the recovery from cDNA of infectious recombinant respiratory syncytial virus (RSV) strain A2 (P. L. Collins, M. G. Hill, E. Camargo, H. Grosfeld, R. M. Chanock, and B. R. Murphy, Proc. Natl. Acad. Sci. USA, 92:11563-11567, 1995). Here, the system was used to construct recombinant RSV containing an additional gene encoding chloramphenicol acetyltransferase (CAT). The CAT coding sequence was flanked by RSV-specific gene-start and gene-end motifs, the transcription signals for the viral RNA-dependent RNA polymerase. The RSV-CAT chimeric transcription cassette was inserted into the region between the G and F genes of the complete cDNA-encoded positive-sense RSV antigenome, and infectious CAT-expressing recombinant RSV was recovered. Transcription of the inserted gene into the predicted subgenomic polyadenylated mRNA was demonstrated by Northern (RNA) blot hybridization analysis, and the encoded protein was detected by enzyme assay and by radioimmunoprecipitation. Quantitation of intracellular CAT, SH, G, and F mRNAs showed that the CAT mRNA was efficiently expressed and that the levels of the G and F mRNAs (which represent the genes on either side of the inserted CAT gene) were comparable to those expressed by a wild-type recombinant RSV. Consistent with this finding, the CAT-containing and wild-type viruses were very similar with regard to the levels of synthesis of the major viral proteins. Each of 25 RSV isolates obtained by plaque purification following eight serial passages expressed CAT, showing that the foreign gene was faithfully maintained in functional form. Analysis by reverse transcription and PCR did not reveal evidence of deletion of the foreign sequence. This finding demonstrated that the RSV genome can accept and maintain an increase in length of 762 nucleotides of foreign sequence and can be engineered to encode an additional, 11th mRNA. The presence of the additional gene resulted in a 10% decrease in plaque diameter and was associated with delay in virus growth and 20-fold decrease in virus yield in vitro. Thus, introduction of an additional gene into the RSV genome might represent a method of attenuation. The ability to express foreign genes by recombinant RSV has implications for basic studies as well as for the development of live recombinant vaccines.
Sustainability in practice: An assessment and advisory platform for everyday use: Experience in the mould&die industry

The spread of the sustainability concept in heterogeneous contexts of use doesn't go together with a similar diffusion of operative instruments supporting the actual implementation of the sustainability paradigm in real manufacturing environments. The here presented SAM (Sustainability Assessment and advisory in Mould&die) platform perfectly fits a traditional product design process, thus enabling product designers and product managers, with limited knowledge on sustainability-related topics, to real-timely ponder the sustainability performances of both existing and under development products. The mould&die industry has been selected for this first deployment.
Molecular Shape, Architecture, and Size of P2X4 Receptors Determined Using Fluorescence Resonance Energy Transfer and Electron Microscopy*

P2X receptors are ATP-gated nonselective cation channels with important physiological roles. However, their structures are poorly understood. Here, we analyzed the architecture of P2X receptors using fluorescence resonance energy transfer (FRET) microscopy and direct structure determination using electron microscopy. FRET efficiency measurements indicated that the distance between the C-terminal tails of P2X4 receptors was 5.6 nm. Single particle analysis of purified P2X4 receptors was used to determine the three-dimensional structure at a resolution of 21Å; the orientation of the particle with respect to the membrane was assigned by labeling the intracellular C termini with 1.8-nm gold particles and the carbohydrate-rich ectodomain with lectin. We found that human P2X4 is a globular torpedo-like molecule with an approximate volume of 270 nm3 and a compact propeller-shaped ectodomain. In this structure, the distance between the centers of the gold particles was 6.1 nm, which closely matches FRET data. Thus, our data provide the first views of the architecture, shape, and size of single P2X receptors, furthering our understanding of this important family of ligand-gated ion channels.
Evaluation of edge detection methods through psychological tests-is the detected edge really desirable for humans?

We propose a method to quantitatively evaluate image processing methods on how well they simulate human visual processing through psychological and statistical tests. First, we focus on edge detection, an image processing technique. To set all edge detection methods fairly, we collect the edge data detected by humans. Then, the signal processing parameters for each edge detection method are adjusted with the same optimizing method based on minimizing the error between the edges detected by the detector and by humans. Two psychological experiments are conducted, confirming whether the edge detection parameters are optimized and evaluating the performance of edge detection methods. The results indicate that the optimizing method functioned comparatively well. The effectiveness rank of edge detection methods is also obtained.
Investigation of a new intervention for children with word-finding problems.

BACKGROUND Around one-quarter of children attending language support services have difficulty in retrieving words. Therapy studies with such children have shown that both semantic and phonological techniques can improve word finding. A new approach to intervention is described using a computerized aid that converts letters into sound cues. AIMS (1) To assess the word-finding abilities in a small series of children with word-finding difficulties. (2) To investigate whether intervention with a computerized aid can improve children's ability to find words. (3) To evaluate the effect of therapy on connected speech (using the Test of Word Finding in Discourse) and in relation to children's, parents' and other practitioners' views (using a questionnaire). (4) To understand the outcome in relation to an interactive model of speech production. METHODS & PROCEDURES Five children (aged 6:10-10:7) were assessed on a variety of background measures. Following baseline assessment, two sets of items were included in therapy: a research set (with matched control items) and a further set selected for their functional relevance for each child. Intervention using the computerized aid took place once a week for half a term (6 weeks). OUTCOMES & RESULTS The children differed from one another in their language development and non-verbal abilities, but all had difficulties in word retrieval. All children showed significant improvement in naming intervention items after therapy. This effect was maintained half a term later. Despite very different profiles, the children benefited similarly from the intervention, suggesting a common processing source for change. After the intervention, two children showed a significant reduction in word-finding behaviours (e.g. use of substitutions, repetitions and empty words) on the Test of Word Finding in Discourse. The views of participants, parents and professionals were varied but generally positive. CONCLUSIONS The main effect of therapy can be understood, in relation to an interactive model of speech production, as strengthening links from meaning to form. Use of the aid in therapy improved word finding for targeted items and this occurred within a clinically realistic timeframe. The finding that the gains in word retrieval maintained suggests that the intervention can be functionally useful if target items are chosen appropriately. Measurable change for two children in connected speech suggests wider benefits can also occur. Overall, the aid can be a useful additional tool for therapists/teachers working with children with this difficulty.
Report of Signal Components Allow for Relative-Distance Assessment

Wouter Halfwerk,1,2,* Rachel A. Page,1 Ryan C. Taylor,3 Preston S. Wilson,4 and Michael J. Ryan1,5 1Smithsonian Tropical Research Institute, PO Box 0843-03092, Balboa, Ancón, Republic of Panama 2Institute of Biology, Leiden University, 2300 RA Leiden, the Netherlands 3Department of Biology, Salisbury University, Salisbury, MD 21801, USA 4Department ofMechanical Engineering, University of Texas at Austin, Austin, TX 78712, USA 5Department of Integrative Biology, University of Texas at Austin, Austin, TX 78712, USA
trans-Dichloridobis(triphenylphosphane-κP)palladium(II) benzene hemisolvate

The title complex, [PdCl2(C18H15P)2]·0.5C6H6, has the PdII ion in a square-planar coordination mode (r.m.s. deviation for Pd, P and Cl atoms = 0.024 Å) with the PPh3 and Cl ligands mutually trans. The benzene solvent molecule is located about a crystallographic inversion centre. The title complex is isostructural with trans-dichloridobis(triphenylphosphane)palladium(II) 1,4-dichlorobenzene sesquisolvate [Kitano et al. (1983 ▶). Acta Cryst. C39, 1015–1017].
Correlates of Self-Concept among Variant Children

48 children who were educably mentally handicapped, learning disabled, achievers, and academically gifted took the Piers-Harris self-concept scale. Differences between groups on the 6 factors of the scale were examined. Achievers and gifted students had higher and more positive responses to self-concept statements about school ability and achievement than the other two groups.
Appraisals, psychotic symptoms and affect in daily life

Background Psychological models of psychosis were examined using Experience Sampling Methods (ESM) to explore relationships between dimensions and appraisals of key symptoms and affect. Method Individuals were signalled to complete ESM booklets 10 times per day for six consecutive days; 534 data points were obtained from 12 out-patients with psychosis. Results Although only 3.6% of spontaneous thoughts were psychosis related, these predicted more negative and less positive affect. Delusions and hallucinations, when present, were rated at a moderate level of intensity, and intensity was associated with distress, interference and preoccupation. Symptom dimensions were related to each other, with weaker associations with delusional conviction, which, it is hypothesized, may represent a separate factor. Conviction and appraisals relating to insight and decentring (‘my problems are something to do with the way my mind works’) were highly variable. Decentring appraisals of delusions, but not insight, were associated with less distress. Appraisals about the power of voices were strong predictors of negative affect and symptom distress. Conclusions This study demonstrates that ESM is a useful methodology to capture ‘online’ variability in psychotic phenomenology and provides evidence supporting cognitive models, which posit that psychotic symptoms are multi-dimensional phenomena, shaped by appraisals that, in turn, predict their emotional and behavioural sequelae.
A 60-GHz CMOS Broadband Receiver With Digital Calibration, 20-to-75-dB Gain, and 5-dB Noise Figure

We present a 60-GHz broadband heterodyne receiver with digital calibration. In the LNA, the low-coupling coefficient transformer-based matching network and the tail switch technique are used to achieve broadband operation and increase the gain tuning range with well-defined gain steps, respectively. In the down-conversion mixer, the inductive peaking technique is employed to extend its bandwidth. In the demodulator, inverter-based <inline-formula> <tex-math notation="LaTeX">$g_{m}$ </tex-math></inline-formula> stages are used to improve its gain and linearity performance. The receiver gain tuning and the I/Q mismatch calibration are achieved by utilizing digital gain control and phase tuning schemes. The measurement shows that the receiver achieves a 20-to-75-dB conversion gain, a 5-dB noise figure (NF), and a good gain flatness performance within the complete 60-GHz band (i.e., 57–66 GHz). Measured with a 60-GHz transmitter, the error vector magnitude (EVM) is better than 10% (−20 dB) in each single channel. Fabricated in a 65-nm CMOS, the receiver occupies a silicon area of 1.9 mm <inline-formula> <tex-math notation="LaTeX">$\times$ </tex-math></inline-formula> 0.7 mm (including all pads) and consumes 102.4-mW dc power.
Observation of Floquet-Bloch States on the Surface of a Topological Insulator

Topological Replicas When a periodic perturbation couples strongly to electrons in a solid, replicas of the original electronic levels are predicted to develop at certain energies—the so-called Floquet-Bloch states. Such conditions can be achieved by shining light on a solid, but the effect is challenging to observe. Wang et al. (p. 453) used time- and angle-resolved photoemission spectroscopy to photoexcite Bi2Se3 and observe its dispersion at various delay times. The replicas were seen at expected energy shifts, along with the gaps predicted to occur at the new energy-level crossings caused by the appearance of the replicas. Because Bi2Se3 is a topological insulator, the breaking of the time-reversal symmetry caused by circularly polarized light resulted in the appearance of an energy gap at the Dirac point, indicating an interesting route toward manipulating electronic states in such materials. Time- and angle-resolved photoemission spectroscopy is used to observe coherent coupling of light with electronic states. The unique electronic properties of the surface electrons in a topological insulator are protected by time-reversal symmetry. Circularly polarized light naturally breaks time-reversal symmetry, which may lead to an exotic surface quantum Hall state. Using time- and angle-resolved photoemission spectroscopy, we show that an intense ultrashort midinfrared pulse with energy below the bulk band gap hybridizes with the surface Dirac fermions of a topological insulator to form Floquet-Bloch bands. These photon-dressed surface bands exhibit polarization-dependent band gaps at avoided crossings. Circularly polarized photons induce an additional gap at the Dirac point, which is a signature of broken time-reversal symmetry on the surface. These observations establish the Floquet-Bloch bands in solids and pave the way for optical manipulation of topological quantum states of matter.
Calcineurin activity assay measurement by liquid chromatography-tandem mass spectrometry in the multiple reaction monitoring mode.

BACKGROUND Blood concentrations of the calcineurin inhibitors (CNIs) cyclosporine and tacrolimus are currently measured to monitor immunosuppression in transplant patients. The measurement of calcineurin (CN) phosphatase activity has been proposed as a complementary pharmacodynamic approach. However, determining CN activity with current methods is not practical. We developed a new method amenable to routine use. METHODS Using liquid chromatography-multiple reaction monitoring mass spectrometry (LC-MRM-MS), we quantified CN activity by measuring the dephosphorylation of a synthetic phosphopeptide substrate. A stable isotope analog of the product peptide served as internal standard, and a novel inhibitor cocktail minimized dephosphorylation by other major serine/threonine phosphatases. The assay was used to determine CN activity in peripheral blood mononuclear cells (PBMCs) isolated from 20 CNI-treated kidney transplant patients and 9 healthy volunteers. RESULTS Linearity was observed from 0.16 to 2.5 μmol/L of product peptide, with accuracy in the 15% tolerance range. Intraassay and interassay recoveries were 100.6 (9.6) and 100 (7.5), respectively. Michaelis-Menten kinetics for purified CN were Km = 10.7 (1.6) μmol/L, Vmax = 2.8 (0.3) μmol/min · mg, and for Jurkat lysate, Km = 182.2 (118.0) μmol/L, Vmax = 0.013 (0.006) μmol/min · mg. PBMC CN activity was successfully measured in a single tube with an inhibitor cocktail. CONCLUSIONS Because LC-MRM-MS is commonly used in routine clinical dosage of drugs, this CN activity assay could be applied, with parallel blood drug concentration monitoring, to a large panel of patients to reevaluate the validity of PBMC CN activity monitoring.
Laboratory framework TEAM for investigating the dependability issues of microprocessor systems

We propose a laboratory research and student training oriented framework consisting of Test Evaluation Automated Means (TEAM) as a set of tools for evaluating the quality of test programs for microprocessors and systems. TEAM enables students to learn and analyze the dependability issues of microprocessor systems, to create their own designs and develop test programs, to analyze the quality of testing, and to make decisions about improving the testability of systems. The tool set in TEAM is mostly open and consists of the assembly level test converter, register transfer level test program simulator, global test converter into local test sequences for modules of the system, and gate-level fault simulator. The general ideas of hands-on laboratory training supported by TEAM are outlined. The research tasks of test program generation can be set up in a way that enables a competition between students, and as a consequence, motivates them to better understand the problems of testing of complex systems and their dependability.
Structural Biology Conformational fl exibility of PL 12 family heparinases : structure and substrate speci fi city of heparinase III from Bacteroides thetaiotaomicron ( BT 4657 )

Glycosaminoglycans (GAGs) are linear polysaccharides comprised of disaccharide repeat units, a hexuronic acid, glucuronic acid or iduronic acid, linked to a hexosamine, N-acetylglucosamine (GlcNAc) or N-acetylgalactosamine. GAGs undergo further modification such as epimerization and sulfation. These polysaccharides are abundant in the extracellular matrix and connective tissues. GAGs function in stabilization of the fibrillar extracellular matrix, control of hydration, regulation of tissue, organism development by controlling cell cycle, cell behavior and differentiation. Niche adapted bacteria express enzymes called polysaccharide lyases (PL), which degrade GAGs for their nutrient content. PL have been classified into 24 sequence-related families. Comparison of 3D structures of the prototypic members of these families allowed identification of distant evolutionary relationships between lyases that were unrecognized at the sequence level, and identified occurrences of convergent evolution. We have characterized structurally and enzymatically heparinase III from Bacteroides thetaiotaomicron (BtHepIII; gene BT4657), which is classified within the PL12 family. BtHepIII is a 72.5 kDa protein. We present the X-ray structures of two crystal forms of BtHepIII at resolution 1.8 and 2.4 Å. BtHepIII contains two domains, the N-terminal α-helical domain forming a toroid and the C-terminal β-sheet domain. Comparison with recently determined structures of two other heparinases from the same PL12 family allowed us to identify structural flexibility in the arrangement of the domains indicating open–close movement. Based on © The Author 2016. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com 176 comparison with other GAG lyases, we identified Tyr301 as the main catalytic residue and confirmed this by site-directed mutagenesis. We have characterized substrate preference of BtHepIII toward sulfate-poor heparan sulfate substrate.
Phonon Mean Free Path in Few Layer Graphene, Hexagonal Boron Nitride, and Composite Bilayer h-BN/Graphene

In this study, ab-initio calculations were performed to obtain the phonon dispersions of seven different structures: single layer graphene, bilayer graphene, graphite, single layer h-BN, bilayer h-BN, bulk h-BN, and finally composite bilayer h-BN/graphene. Using these dispersions specific heat, group velocity, and single mode relaxation times of phonons were obtained to calculate their thermal conductivities, and mean free paths at room temperature. Calculated variables were used to understand the effects of additional layers to thermophysical properties, phonon mode contributions to thermal conductivity, and the limits for ballistic-diffusive heat transfer of all seven structures. Finally, based on the obtained data, thermal properties of the composite bilayer h-BN/graphene were investigated and compared to those of few layer graphene and h-BN structures.
The history of the formation of Deaf Children's Theatre in the United States

THE HISTORY OF THE FORMATION OF DEAF CHILDREN’S THEATRE IN THE UNITED STATES BRIAN R. KILPATRICK This dissertation explores the history of Deaf Children’s Theatre in America, by collecting and recording its formation and development from archival materials, from on-site interviews, and from documents published during the late twentieth century. This study investigates the ways in which theatre became accessible to deaf people through apposite legislation and describes several forms of accessibility services for deaf and hearing audiences at theatrical performances. The evolution of theatre leading to the formation of Deaf Theatre and, later, to the nascence of Deaf Children’s Theatre is explored within the larger social and political context of the American culture. The case history of each of the six Deaf Children’s Theatre groups established variously at Hartford, Connecticut; Bethesda, Maryland; Seattle, Washington; Boston, Massachusetts; Chicago, Illinois; and Houston, Texas is presented to portray the particular development of Theatre Arts for deaf children in America. The six Deaf Children’s Theatre groups presented in this study have had different approaches to complementing and completing deaf children’s education. Suggested future research is directed to the potential future impact of drama on deaf children’s language development and also to various avenues of support for Deaf Theatre in its battle for survival.
Measures taken to reduce the risk of West Nile virus transmission by transplantation in Italy.

For the first time in Italy, two patients with meningoencephalitis were diagnosed with West Nile virus (WNV) infection in September 2008 [1]. The patients live in the Bologna and Ferrara provinces of Emilia Romagna where WNV infections had previously been noted in horses, crows and magpies [2]. The Italian National Transplant Centre (CNT), which is responsible for the procurement, processing and distribution of organs and tissues in Italy, has now reviewed the risks of transmission of WNV by organ, tissue and cell transplantation and, taking into account the advice and recommendations of the relevant authorities in other countries, issued guidance to the transplant community.