Arp2p, a potential substrate for NatB acetyltransferase in fission yeast, is important in actin patch nucleation and motility in arp2-1 and arm1 ts mutants

The actin cytoskeleton is a dynamic and complex structure in fission yeast that plays a major function in many cell processes including cellular growth, septa formation, endocytosis and cellular division. Computational studies have shown that Arp2p, which forms part of the Arp2/3 complex, is a potential substrate of NatB acetyltransferase which has specificity for proteins possessing an N-terminal Met-Asp or Met-Glu sequence motif. In arm1- mutants the loss of function of Arm1p, an auxillary subunit required for NatB activity, results in a temperature sensitive phenotype characterized by multiple septa, failure of endocytosis, and the inability to form actin cables. A temperature sensitive mutant of Schizosaccharomyces pombe arp2 gene exhibits a similar phenotype as seen by the formation of improper septa, slow growth, and the delocalization of actin patches. Four expression vectors encoding the open reading frames of arp2 and cdc8 (tropomyosin) were constructed with a modification changing the second residue to a Histidine, believed to mimic the charge distribution of natural acetylation by NatB. Constructs tested in normal yeast strains remained viable and grew normally in the presence of Met-His Arp2p and tropomyosin. Analysis of their ability to suppress the mutant phenotypes of arp2-1 and arm1- mutants is an area of research to be explored in future studies.

Continuity - based and discontinuity - based segmentation in transparent and spatially segregated global motion

The mechanisms underlying the parsing of a spatial distribution of velocity vectors into two adjacent (spatially segregated) or overlapping (transparent) motion surfaces were examined using random dot kinematograms. Parsing might occur using either of two principles. Surfaces might be defined on the basis of similarity of motion vectors and then sharp perceptual boundaries drawn between different surfaces (continuity-based segmentation). Alternatively, detection of a high gradient of direction or speed separating the motion surfaces might drive the process (discontinuity-based segmentation). To establish which method is used, we examined the effect of blurring the motion direction gradient. In the case of a sharp direction gradient, each dot had one of two directions differing by 135°. With a shallow gradient, most dots had one of two directions but the directions of the remainder spanned the range between one motion-defined surface and the other. In the spatial segregation case the gradient defined a central boundary separating two regions. In the transparent version the dots were randomly positioned. In both cases all dots moved with the same speed and existed for only two frames before being randomly replaced. The ability of observers to parse the motion distribution was measured in terms of their ability to discriminate the direction of one of the two surfaces. Performance was hardly affected by spreading the gradient over at least 25% of the dots (corresponding to a 1° strip in the segregation case). We conclude that detection of sharp velocity gradients is not necessary for distinguishing different motion surfaces.

Amphibian skin peptides and their corresponding cDNAs from single lyophilized secretion samples: identification of novel brevinins from three species of Chinese frogs

Brevinins are peptides of 24 amino acid residues, originally isolated from the skin of the Oriental frog, Rana brevipoda porsa, by nature of their microbicidal activity against a wide range of Gram-positive and Gram-negative bacteria and against strains of pathogenic fungi. cDNA libraries were constructed from lyophilized skin secretion of three, unstudied species of Chinese frog, Odorrana schmackeri, Odorrana versabilis and Pelophylax plancyi fukienensis, using our recently developed technique. In this report, we describe the “shotgun” cloning of novel brevinins by means of 3'-RACE, using a “universal” degenerate primer directed towards a highly conserved nucleic acid sequence domain within the 5'-untranslated region of previously characterized frog skin peptide cDNAs. Novel brevinins, deduced from cloned cDNA open-reading frames, were subsequently identified as mature peptides in the same samples of respective species skin secretions. Bioinformatic analysis of both prepro-brevinin nucleic acid sequences and translated open-reading frame amino acid sequences revealed a highly conserved signal peptide domain and a hypervariable anti-microbial peptide-encoding domain. The experimental approach described here can thus rapidly provide robust structural data on skin anti-microbial peptides without harming the donor amphibians.

Cultural policy models and cultural facilities models: from national to local models. Analysis of Barcelona’s case

The development of cultural policy analysis by social science has been produced a theorization about cultural policy models from sociology and political science. This analysis shows the influence of the national model of cultural policy on the forms of governance and management of cultural facilities. However, in this paper we will defend that currently the local model of cultural policy decisively influences the model of cultural institutions. This is explained by the growing importance of culture in local development strategies. In order to demonstrate this we will analyze the case of the Barcelona Model of local development and cultural policy, that is characterized for the level of local government leadership, multilevel governance, the use of culture in urban planning processes and a tendency to use public-private partnership in public management. This Model influences the genesis and development of the cultural facilities and it produces a singular and relatively successful model.

Novel frenatins from the skin of the Australasian Giant White-lipped tree frog, Litoria infrafrenata: cloning of precursor cDNAs and identification in defensive skin secretion.

The Australasian anuran amphibian genus Litoria, contains many phenotypically-diverse species as a result of radial evolution of an ancestral species into different biotopes much in the manner of the indigenous marsupial mammals. In common with members of the Central/South American genus Phyllomedusa, their specialized skin granular glands are factories for the production of a plethora of biologically-active peptides. Here we report a more detailed study of those present in the defensive skin secretion of the Australasian giant white-lipped tree frog, Litoria infrafrenata, and, for the first time, we have identified three novel frenatins by deduction of primary structures from cDNAs that were cloned from a library constructed from lyophilized skin secretion using a recently-developed technique. All open-reading frames consisted of a putative signal peptide and an acidic pro-region followed by a single copy of a frenatin peptide. Processed peptides corresponding in molecular mass to the deduced molecular masses of frenatins (named 1.1, 3, 3.1 and 4.1) were identified in the same secretion sample using HPLC and mass spectroscopy. The application of this technique thus permits parallel peptidomic and transcriptomic analyzes on the same lyophilized skin secretion sample circumventing sacrifice of specimens from endangered herpetofauna.

The structural organization of aurein precursor cDNAs from the skin secretion of the Australian green and golden bell frog, Litoria aurea.

Aureins are a family of peptides (13-25 residues), some of which possess potent antimicrobial and anti-cancer properties, which have been classified into 5 subgroups based upon primary structural similarities. They were originally isolated from the defensive skin secretions of the closely related Australian bell frogs, Litoria aurea and Litoria raniformis, and of the 23 aurein peptides identified, 10 are common to both species. Using a recently developed technique, we have constructed a cDNA library from the defensive secretion of the green and golden bell frog, L. aurea, and successfully cloned a range of aurein precursor transcripts containing entire open-reading frames. All open-reading frames consisted of a putative signal peptide and an acidic pro-region followed by a single copy of aurein. The deduced precursor structures for the most active aureins (2.2 and 3.1) confirmed the presence of a C-terminal amidation motif whereas that of aurein 5.3 did not. Processed peptides corresponding in molecular mass to aureins 2.2, 2.3, 2.5, 3.1 and 5.3 were identified in the same secretion sample using LC/MS. The application of this technique thus permits parallel peptidomic and transcriptomic analyses on the same lyophilized skin secretion sample circumventing sacrifice of specimens of endangered herpetofauna.

Object tracking using SIFT features and mean shift

A scale invariant feature transform (SIFT) based mean shift algorithm is presented for object tracking in real scenarios. SIFT features are used to correspond the region of interests across frames. Meanwhile, mean shift is applied to conduct similarity search via color histograms. The probability distributions from these two measurements are evaluated in an expectation–maximization scheme so as to achieve maximum likelihood estimation of similar regions. This mutual support mechanism can lead to consistent tracking performance if one of the two measurements becomes unstable. Experimental work demonstrates that the proposed mean shift/SIFT strategy improves the tracking performance of the classical mean shift and SIFT tracking algorithms in complicated real scenarios.

Novel loading tests on full-scale tapered member portal frames

This paper presents the experimental results of loading tests on two 18m span tapered member portal frames designed to BS 5950. Deflection test results for vertical, lateral and combined loading cases are compared with the predictions given by elastic analysis to BS 5950 and shown to be favourable. The predicted ultimate capacities and modes of failure, which were by lateral-torsional buckling of the columns, were also found to agree with the experimental behaviour. It was found that the method of modelling the tapered members as a series of prismatic elements gave good comparison with test results.

The circle in the relationship between space and body. Photo-essay based on Isidro Blasco and the BAPNE method

This article demonstrates a visual study on the educational space in which the teaching of body percussion is carried out in universities. The methodological framework is chosen by the Visual Arts Based Educational Research, using the work of an artist as a conceptual and methodological model. The research remains notable (1) due to the theoretical reference to the BAPNE method, (2) due to the visual reference to the work of Isidro Blasco – especially with the piece “Shanghai Planet 2009” - ;(3) due to the parallelisms established between the object of study in this investigation -the spatial analysis- and the focuses of interest revealed by the art critics in relation to the work of this artist. By means of a visual speech formed with 7 photo-collages the relationship between body and educational space is visualized in the basic disposition of circular learning. The visual constructions by way of photo-collage and their aesthetic charge brings us closer to the intimacy of the educational space, in the style in which it is distributed to the students in the music classroom, the materialization of interpersonal relationships, the occupied and empty volumes.

Creating replicas of scultural heritage through 3D reconstruction and low cost 3D printer in Education

The process of making replicas of heritage has traditionally been developed by public agencies, corporations and museums and is not commonly used in schools. Currently there are technologies that allow creating cheap replicas. The new 3D reconstruction software, based on photographs and low cost 3D printers allow to make replicas at a cost much lower than traditional. This article describes the process of creating replicas of the sculpture Goslar Warrior of artist Henry Moore, located in Santa Cruz de Tenerife. To make this process, first, a digital model have been created using Autodesk Recap 360, Autodesk 123D Catch and Autodesk Meshmixer MarkerBot MakerWare applications. Physical replication, has been reproduced in polylactic acid (PLA) by MakerBot Replicator 2 3D printer. In addition, a cost analysis using, in one hand, the printer mentioned, and in the other hand, 3D printing services both online and local, is included. Finally, there has been a specific action with 141 students and 12 high school teachers, who filled a questionnary about the use of sculptural replicas in education.

Hylambatin and (Thr)11-hylambatin from the skin secretion of the African hyperoliid frog, Kassina maculata: Structural characterisation, precursor cDNA cloning and preliminary pharmacological testing

The tachykinins hylambatin and (Thr)11-hylambatin have been isolated from the defensive skin secretion of the African hyperoliid frog, Kassina maculata,. Hylambatin (DPPDPNRFYGMMamide) is revised in structure from the original sequence by a single site substitution (Asn/Asp at position 6), and (Thr)11-hylambatin, a novel tachykinin, differs in structure from hylambatin by a single Thr/Met substitution. (Thr)11-hylambatin is five- to ten-fold more abundant than hylambatin in secretions. Synthetic replicates of both peptides were active in smooth muscle preparations including the rat tail artery, rat ileum and bovine trachea. While hylambatin displayed activity consistent with an NK1-receptor ligand, (Thr)11-hylambatin was more active than either substance P or neurokinin A in both NK1- and NK-2 receptor rich preparations. Incorporation of a threoninyl residue rather than the canonical leucyl residue at the penultimate position in both substance P and neurokinin A, generated active ligands in both arterial and intestinal smooth muscle preparations. Hylambatin precursor cDNAs, designated HYBN-1 and HYBN-2, respectively, were cloned from a skin library by 3'- and 5'-RACE reactions. Both were highly-homologous containing open-reading frames of 66 amino acids encoding single copies of either hylambatin or (Thr)11-hylambatin. These data reveal a hitherto unrecognized structure/activity attribute of mammalian tachykinin receptors revealed though discovery of a novel amphibian skin-derived, site-substituted peptide ligand.

Spatial Perception and Cognition in Multichannel Audio for Electroacoustic Music

Listeners experience electroacoustic music as full of significance and meaning, and they experience spatiality as one of the factors contributing to its meaningfulness. If we want to understand spatiality in electroacoustic music, we must understand how the listener’s mental processes give rise to the experience of meaning. In electroacoustic music as in everyday life, these mental processes unite the peripheral auditory system with human spatial cognition. In the discussion that follows we consider a range of the listener’s mental processes relating space and meaning from the perceptual attributes of spatial imagery to the spatial reference frames for places and navigation. When considering multichannel loudspeaker systems in particular, an important part of the discussion is focused on the distinctive and idiomatic ways in which this particular mode of sound production contributes to and situates meaning. These idiosyncrasies include the phenomenon of image dispersion, the important consequences of the precedence effect and the influence of source characteristics on spatial imagery. These are discussed in close relation to the practicalities of artistic practice and to the potential for artistic meaning experienced by the listener.

On-line Emotion Recognition in a 3-D Activation-Valence-Time Continuum using Acoustic and Linguistic Cues

For many applications of emotion recognition, such as virtual agents, the system must select responses while the user is speaking. This requires reliable on-line recognition of the user’s affect. However most emotion recognition systems are based on turnwise processing. We present a novel approach to on-line emotion recognition from speech using Long Short-Term Memory Recurrent Neural Networks. Emotion is recognised frame-wise in a two-dimensional valence-activation continuum. In contrast to current state-of-the-art approaches, recognition is performed on low-level signal frames, similar to those used for speech recognition. No statistical functionals are applied to low-level feature contours. Framing at a higher level is therefore unnecessary and regression outputs can be produced in real-time for every low-level input frame. We also investigate the benefits of including linguistic features on the signal frame level obtained by a keyword spotter.

Kassorins: Novel innate immune system peptides from skin secretions of the African hyperoliid frogs, Kassina maculata and Kassina senegalensis

From defensive skin secretions acquired from two species of African hyperoliid frogs, Kassina maculata and Kassina senegalensis, we have isolated two structurally related, C-terminally amidated tridecapeptides of novel primary structure that exhibit a broad spectrum of biological activity. In reflection of their structural novelty and species of origin, we named the peptides kassorin M (FLEGLLNTVTGLLamide; 1387.8 Da) and kassorin S (FLGGILNTITGLLamide; 1329.8 Da), respectively. The primary structure and organisation of the biosynthetic precursors of kassorins M and S were deduced from cloned skin secretion-derived cDNA. Both open-reading frames encoded a single copy of kassorin M and S, respectively, located at the C-terminus. Kassorins display limited structural similarities to vespid chemotactic peptides (7/13 residues), temporin A (5/13 residues), the N-terminus of Lv-ranaspumin, a foam nest surfactant protein of the frog, Leptodactylus vastus, and an N-terminal domain of the equine sweat surfactant protein, latherin. Both peptides elicit histamine release from rat peritoneal mast cells. However, while kassorin S was found to possess antibacterial activity against Staphylococcus aureus, kassorin M was devoid of such activity. In contrast, kassorin M was found to contract the smooth muscle of guinea pig urinary bladder (EC50 = 4.66 nM) and kassorin S was devoid of this activity. Kassorins thus represent the prototypes of a novel family of peptides from the amphibian innate immune system as occurring in defensive skin secretions.

Effect of column base strength on steel portal frames in fire

This paper presents the findings of a project part sponsored by an ICE Research and Development grant on portal frames in fire. The research reported here has also lead to a sucessful research grant from the IStructE. The paper describes a non-linear elasto plastic dynamic finite element model that captures the collapse of a portal frame in fire. It demonstrates that current guidance on the column base stiffness and strength, to prevent collapse, may in some cases be unconservative.