Exploitation of Ebola Virus VP35 Protein to identify new drugs counteracting its type I IFN antagonism

Ebolaviruses (EBOVs) are among the most virulent and deadly pathogens ever known, causing fulminant haemorrhagic fevers in humans and non-human primates. The 2014 outbreak of Ebola virus disease (EVD) in West Africa has claimed more lives than all previous EVD outbreaks combined. The EBOV high mortality rates have been related to the virus-induced impairment of the host innate immunity reaction due to two virus-coded proteins, VP24 and VP35. EBOV VP35 is a multifunctional protein, it is essential for viral replication as a component of the viral RNA polymerase and it also participates in nucleocapsid assembly. Early during EBOV infection, alpha-beta interferon (IFN-α/β) production would be triggered upon recognition of viral dsRNA products by cytoplasmic retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs). However, this recognition is efficiently prevented by the double-stranded RNA (dsRNA) binding activity of the EBOV VP35 protein, which hides RLRs binding sites on the dsRNA phosphate backbone as well the 5’-triphosphate (5’-ppp) dsRNA ends to RIG-I recognition. In addition to dsRNA binding and sequestration, EBOV VP35 inhibits IFN-α/β production preventing the activation of the IFN regulatory factor 3 (IRF-3) by direct interaction with cellular proteins. Previous studies demonstrated that single amino acid changes in the VP35 dsRNA binding domain reduce EBOV virulence, indicating that VP35 is an attractive target for antiviral drugs development. Within this context, here we report the establishment of a novel method to characterize the EBOV VP35 inhibitory function of the dsRNA-dependent RIG-I-mediated IFN-β signaling pathway in a BLS2 cell culture setting. In such system, a plasmid containing the promoter region of IFN-β gene linked with a luciferase reporter gene was transfected, together with a EBOV VP35 mammalian expression plasmid, into the IFN-sensitive A549 cell line, and the IFN-induction was stimulated through dsRNA transfection. Through alanine scanning mutational studies with biochemical, cellular and computational methods we highlighted the importance of some VP35 residues involved in dsRNA end-capping binding, such as R312, K282 and R322, that may serve as target for the development of small-molecule inhibitors against EBOV. Furthermore, we identified a synthetic compound that increased IFN-induction only under antiviral response stimulation and subverted VP35 inhibition, proving to be very attractive for the development of an antiviral drug. In conclusion, our results provide the establishment of a new assay as a straightforward tool for the screening of antiviral compounds that target i) dsRNA-VP35 or cellular protein-VP35 interaction and ii) dsRNA-dependent RIG-I-mediated IFN signaling pathway, in order to potentiate the IFN response against VP35 inhibition, setting the bases for further drug development.

Information sets and partial permutation decoding for codes from finite geometries

Mavron, Vassili; McDonough, T.P.; Key, J.D., (2006) 'Information sets and partial permutation decoding for codes from finite geometries', Finite Fields and their applications 12(2) pp.232-247 RAE2008

Experimental and numerical simulation of dough kneading in filled geometries

Binding, David; Couch, M.A.; Sujatha, K.S.; Webster, M.F., (2003) 'Experimental and numerical simulation of dough kneading in filled geometries', Journal of Food Engineering 58 pp.111-123 RAE2008

Some preliminary conjunctural thoughts on countercultures

The article is devoted to analysis of various countercultures aspects. The Author hypothesizes that the forms of contemporary politics are the result of two fundamentally interrelated strategic vectors. First is built upon a politics of ressentiment, setting “ordinary” folks against the socio‐cultural elite which constututed to the “new right” movement. Second vector is originated of counterculture of the 1960s. The author states that counterculture exists without a singular identity. It is a space of hybridity and heterogenity. On the other hand counterculture is related to the concept of contemporary ambivalence as well it transforms of affective experience of everyday life. Another feature of counterculture is connected with the conviction that counterculture stands against of dominant culture. Also the author puts counterculture movements against various aspects of new american modernity including reconstruction of the practice of the hegemony, as well as through popular culture and reconstruction of the „left” and „right” idelogy and practice.

Vírus ao serviço da indústria farmacêutica

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

Anestésicos locais

Projeto de Pós-Graduação/Dissertação apresentado à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Mestre em Ciências Farmacêuticas

Simultaneous Localization and Recognition of Dynamic Hand Gestures

A framework for the simultaneous localization and recognition of dynamic hand gestures is proposed. At the core of this framework is a dynamic space-time warping (DSTW) algorithm, that aligns a pair of query and model gestures in both space and time. For every frame of the query sequence, feature detectors generate multiple hand region candidates. Dynamic programming is then used to compute both a global matching cost, which is used to recognize the query gesture, and a warping path, which aligns the query and model sequences in time, and also finds the best hand candidate region in every query frame. The proposed framework includes translation invariant recognition of gestures, a desirable property for many HCI systems. The performance of the approach is evaluated on a dataset of hand signed digits gestured by people wearing short sleeve shirts, in front of a background containing other non-hand skin-colored objects. The algorithm simultaneously localizes the gesturing hand and recognizes the hand-signed digit. Although DSTW is illustrated in a gesture recognition setting, the proposed algorithm is a general method for matching time series, that allows for multiple candidate feature vectors to be extracted at each time step.

Indexing Methods for Efficient Multiclass Recognition

Many real world image analysis problems, such as face recognition and hand pose estimation, involve recognizing a large number of classes of objects or shapes. Large margin methods, such as AdaBoost and Support Vector Machines (SVMs), often provide competitive accuracy rates, but at the cost of evaluating a large number of binary classifiers, thus making it difficult to apply such methods when thousands or millions of classes need to be recognized. This thesis proposes a filter-and-refine framework, whereby, given a test pattern, a small number of candidate classes can be identified efficiently at the filter step, and computationally expensive large margin classifiers are used to evaluate these candidates at the refine step. Two different filtering methods are proposed, ClassMap and OVA-VS (One-vs.-All classification using Vector Search). ClassMap is an embedding-based method, works for both boosted classifiers and SVMs, and tends to map the patterns and their associated classes close to each other in a vector space. OVA-VS maps OVA classifiers and test patterns to vectors based on the weights and outputs of weak classifiers of the boosting scheme. At runtime, finding the strongest-responding OVA classifier becomes a classical vector search problem, where well-known methods can be used to gain efficiency. In our experiments, the proposed methods achieve significant speed-ups, in some cases up to two orders of magnitude, compared to exhaustive evaluation of all OVA classifiers. This was achieved in hand pose recognition and face recognition systems where the number of classes ranges from 535 to 48,600.

Combining Textual and Visual Cues for Content-based Image Retrieval on the World Wide Web

Some WWW image engines allow the user to form a query in terms of text keywords. To build the image index, keywords are extracted heuristically from HTML documents containing each image, and/or from the image URL and file headers. Unfortunately, text-based image engines have merely retro-fitted standard SQL database query methods, and it is difficult to include images cues within such a framework. On the other hand, visual statistics (e.g., color histograms) are often insufficient for helping users find desired images in a vast WWW index. By truly unifying textual and visual statistics, one would expect to get better results than either used separately. In this paper, we propose an approach that allows the combination of visual statistics with textual statistics in the vector space representation commonly used in query by image content systems. Text statistics are captured in vector form using latent semantic indexing (LSI). The LSI index for an HTML document is then associated with each of the images contained therein. Visual statistics (e.g., color, orientedness) are also computed for each image. The LSI and visual statistic vectors are then combined into a single index vector that can be used for content-based search of the resulting image database. By using an integrated approach, we are able to take advantage of possible statistical couplings between the topic of the document (latent semantic content) and the contents of images (visual statistics). This allows improved performance in conducting content-based search. This approach has been implemented in a WWW image search engine prototype.

Combinations of Deformable Shape Prototypes

We propose to investigate a model-based technique for encoding non-rigid object classes in terms of object prototypes. Objects from the same class can be parameterized by identifying shape and appearance invariants of the class to devise low-level representations. The approach presented here creates a flexible model for an object class from a set of prototypes. This model is then used to estimate the parameters of low-level representation of novel objects as combinations of the prototype parameters. Variations in the object shape are modeled as non-rigid deformations. Appearance variations are modeled as intensity variations. In the training phase, the system is presented with several example prototype images. These prototype images are registered to a reference image by a finite element-based technique called Active Blobs. The deformations of the finite element model to register a prototype image with the reference image provide the shape description or shape vector for the prototype. The shape vector for each prototype, is then used to warp the prototype image onto the reference image and obtain the corresponding texture vector. The prototype texture vectors, being warped onto the same reference image have a pixel by pixel correspondence with each other and hence are "shape normalized". Given sufficient number of prototypes that exhibit appropriate in-class variations, the shape and the texture vectors define a linear prototype subspace that spans the object class. Each prototype is a vector in this subspace. The matching phase involves the estimation of a set of combination parameters for synthesis of the novel object by combining the prototype shape and texture vectors. The strengths of this technique lie in the combined estimation of both shape and appearance parameters. This is in contrast with the previous approaches where shape and appearance parameters were estimated separately.

Fuzzy ARTMAP, Slow Learning and Probability Estimation

A nonparametric probability estimation procedure using the fuzzy ARTMAP neural network is here described. Because the procedure does not make a priori assumptions about underlying probability distributions, it yields accurate estimates on a wide variety of prediction tasks. Fuzzy ARTMAP is used to perform probability estimation in two different modes. In a 'slow-learning' mode, input-output associations change slowly, with the strength of each association computing a conditional probability estimate. In 'max-nodes' mode, a fixed number of categories are coded during an initial fast learning interval, and weights are then tuned by slow learning. Simulations illustrate system performance on tasks in which various numbers of clusters in the set of input vectors mapped to a given class.

Neural Representations for Sensory-Motor Control, III: Learning a Body-Centered Representation of 3-D Target Position

A neural model is described of how the brain may autonomously learn a body-centered representation of 3-D target position by combining information about retinal target position, eye position, and head position in real time. Such a body-centered spatial representation enables accurate movement commands to the limbs to be generated despite changes in the spatial relationships between the eyes, head, body, and limbs through time. The model learns a vector representation--otherwise known as a parcellated distributed representation--of target vergence with respect to the two eyes, and of the horizontal and vertical spherical angles of the target with respect to a cyclopean egocenter. Such a vergence-spherical representation has been reported in the caudal midbrain and medulla of the frog, as well as in psychophysical movement studies in humans. A head-centered vergence-spherical representation of foveated target position can be generated by two stages of opponent processing that combine corollary discharges of outflow movement signals to the two eyes. Sums and differences of opponent signals define angular and vergence coordinates, respectively. The head-centered representation interacts with a binocular visual representation of non-foveated target position to learn a visuomotor representation of both foveated and non-foveated target position that is capable of commanding yoked eye movementes. This head-centered vector representation also interacts with representations of neck movement commands to learn a body-centered estimate of target position that is capable of commanding coordinated arm movements. Learning occurs during head movements made while gaze remains fixed on a foveated target. An initial estimate is stored and a VOR-mediated gating signal prevents the stored estimate from being reset during a gaze-maintaining head movement. As the head moves, new estimates arc compared with the stored estimate to compute difference vectors which act as error signals that drive the learning process, as well as control the on-line merging of multimodal information.

A Neural Network Architecture for Autonomous Learning, Recognition, and Prediction in a Nonstationary World

In a constantly changing world, humans are adapted to alternate routinely between attending to familiar objects and testing hypotheses about novel ones. We can rapidly learn to recognize and narne novel objects without unselectively disrupting our memories of familiar ones. We can notice fine details that differentiate nearly identical objects and generalize across broad classes of dissimilar objects. This chapter describes a class of self-organizing neural network architectures--called ARTMAP-- that are capable of fast, yet stable, on-line recognition learning, hypothesis testing, and naming in response to an arbitrary stream of input patterns (Carpenter, Grossberg, Markuzon, Reynolds, and Rosen, 1992; Carpenter, Grossberg, and Reynolds, 1991). The intrinsic stability of ARTMAP allows the system to learn incrementally for an unlimited period of time. System stability properties can be traced to the structure of its learned memories, which encode clusters of attended features into its recognition categories, rather than slow averages of category inputs. The level of detail in the learned attentional focus is determined moment-by-moment, depending on predictive success: an error due to over-generalization automatically focuses attention on additional input details enough of which are learned in a new recognition category so that the predictive error will not be repeated. An ARTMAP system creates an evolving map between a variable number of learned categories that compress one feature space (e.g., visual features) to learned categories of another feature space (e.g., auditory features). Input vectors can be either binary or analog. Computational properties of the networks enable them to perform significantly better in benchmark studies than alternative machine learning, genetic algorithm, or neural network models. Some of the critical problems that challenge and constrain any such autonomous learning system will next be illustrated. Design principles that work together to solve these problems are then outlined. These principles are realized in the ARTMAP architecture, which is specified as an algorithm. Finally, ARTMAP dynamics are illustrated by means of a series of benchmark simulations.

ARTMAP: Supervised Real-Time Learning and Classification of Nonstationary Data by a Self-Organizing Neural Network

This article introduces a new neural network architecture, called ARTMAP, that autonomously learns to classify arbitrarily many, arbitrarily ordered vectors into recognition categories based on predictive success. This supervised learning system is built up from a pair of Adaptive Resonance Theory modules (ARTa and ARTb) that are capable of self-organizing stable recognition categories in response to arbitrary sequences of input patterns. During training trials, the ARTa module receives a stream {a^(p)} of input patterns, and ARTb receives a stream {b^(p)} of input patterns, where b^(p) is the correct prediction given a^(p). These ART modules are linked by an associative learning network and an internal controller that ensures autonomous system operation in real time. During test trials, the remaining patterns a^(p) are presented without b^(p), and their predictions at ARTb are compared with b^(p). Tested on a benchmark machine learning database in both on-line and off-line simulations, the ARTMAP system learns orders of magnitude more quickly, efficiently, and accurately than alternative algorithms, and achieves 100% accuracy after training on less than half the input patterns in the database. It achieves these properties by using an internal controller that conjointly maximizes predictive generalization and minimizes predictive error by linking predictive success to category size on a trial-by-trial basis, using only local operations. This computation increases the vigilance parameter ρa of ARTa by the minimal amount needed to correct a predictive error at ARTb· Parameter ρa calibrates the minimum confidence that ARTa must have in a category, or hypothesis, activated by an input a^(p) in order for ARTa to accept that category, rather than search for a better one through an automatically controlled process of hypothesis testing. Parameter ρa is compared with the degree of match between a^(p) and the top-down learned expectation, or prototype, that is read-out subsequent to activation of an ARTa category. Search occurs if the degree of match is less than ρa. ARTMAP is hereby a type of self-organizing expert system that calibrates the selectivity of its hypotheses based upon predictive success. As a result, rare but important events can be quickly and sharply distinguished even if they are similar to frequent events with different consequences. Between input trials ρa relaxes to a baseline vigilance pa When ρa is large, the system runs in a conservative mode, wherein predictions are made only if the system is confident of the outcome. Very few false-alarm errors then occur at any stage of learning, yet the system reaches asymptote with no loss of speed. Because ARTMAP learning is self stabilizing, it can continue learning one or more databases, without degrading its corpus of memories, until its full memory capacity is utilized.

Vector Associative Maps: Unsupervised Real-time Error-based Learning and Control of Movement Trajectories

This article describes neural network models for adaptive control of arm movement trajectories during visually guided reaching and, more generally, a framework for unsupervised real-time error-based learning. The models clarify how a child, or untrained robot, can learn to reach for objects that it sees. Piaget has provided basic insights with his concept of a circular reaction: As an infant makes internally generated movements of its hand, the eyes automatically follow this motion. A transformation is learned between the visual representation of hand position and the motor representation of hand position. Learning of this transformation eventually enables the child to accurately reach for visually detected targets. Grossberg and Kuperstein have shown how the eye movement system can use visual error signals to correct movement parameters via cerebellar learning. Here it is shown how endogenously generated arm movements lead to adaptive tuning of arm control parameters. These movements also activate the target position representations that are used to learn the visuo-motor transformation that controls visually guided reaching. The AVITE model presented here is an adaptive neural circuit based on the Vector Integration to Endpoint (VITE) model for arm and speech trajectory generation of Bullock and Grossberg. In the VITE model, a Target Position Command (TPC) represents the location of the desired target. The Present Position Command (PPC) encodes the present hand-arm configuration. The Difference Vector (DV) population continuously.computes the difference between the PPC and the TPC. A speed-controlling GO signal multiplies DV output. The PPC integrates the (DV)·(GO) product and generates an outflow command to the arm. Integration at the PPC continues at a rate dependent on GO signal size until the DV reaches zero, at which time the PPC equals the TPC. The AVITE model explains how self-consistent TPC and PPC coordinates are autonomously generated and learned. Learning of AVITE parameters is regulated by activation of a self-regulating Endogenous Random Generator (ERG) of training vectors. Each vector is integrated at the PPC, giving rise to a movement command. The generation of each vector induces a complementary postural phase during which ERG output stops and learning occurs. Then a new vector is generated and the cycle is repeated. This cyclic, biphasic behavior is controlled by a specialized gated dipole circuit. ERG output autonomously stops in such a way that, across trials, a broad sample of workspace target positions is generated. When the ERG shuts off, a modulator gate opens, copying the PPC into the TPC. Learning of a transformation from TPC to PPC occurs using the DV as an error signal that is zeroed due to learning. This learning scheme is called a Vector Associative Map, or VAM. The VAM model is a general-purpose device for autonomous real-time error-based learning and performance of associative maps. The DV stage serves the dual function of reading out new TPCs during performance and reading in new adaptive weights during learning, without a disruption of real-time operation. YAMs thus provide an on-line unsupervised alternative to the off-line properties of supervised error-correction learning algorithms. YAMs and VAM cascades for learning motor-to-motor and spatial-to-motor maps are described. YAM models and Adaptive Resonance Theory (ART) models exhibit complementary matching, learning, and performance properties that together provide a foundation for designing a total sensory-cognitive and cognitive-motor autonomous system.