956 resultados para ENCODE GASP
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bbd18 is a differentially expressed Borrelia burgdorferi gene that is transcribed at almost undetectable levels in spirochetes grown in vitro but dramatically upregulated during tick infection. The gene also displays low yet detectable expression at various times in tissues of murine hosts. As the gene product bears no homology to known proteins, its biological significance remains enigmatic. To understand the gene function, we created isogenic bbd18-deletion mutants as well as genetically-complemented isolates from an infectious wild-type B. burgdorferi strain. Compared to parental isolates, bbd18 mutants - but not complemented spirochetes - displayed slower in vitro growth. The bbd18 mutants also reflect significantly reduced ability to persist or remain undetectable both in immunocompetent and SCID mice, yet were able to survive in ticks. This suggests BBD18 function is essential in mammalian hosts but redundant in the arthropod vector. Notably, although bbd18 expression and in vitro growth defects are restored in the complemented isolates, their phenotype is similar to the mutants - being unable to persist in mice but able to survive in ticks. Despite low expression in cultured wild-type B. burgdorferi, bbd18 deletion downregulated several genes. Interestingly, expression of some, including ospD and bbi39, could be complemented, while that of others could not be restored via bbd18 re-expression. Correspondingly, bbd18 mutants displayed altered production of several proteins, and similar to RNA levels, some were restored in the bbd18 complement and others not. To understand how bbd18 deletion results in apparently permanent and noncomplementable phenotypic defects, we sought to genetically disturb the DNA topology surrounding the bbd18 locus without deleting the gene. Spirochetes with an antibiotic cassette inserted downstream of the gene, between bbd17 and bbd18, were significantly attenuated in mice, while a similar upstream insertion, between bbd18 and bbd19, did not affect infectivity, suggesting that an unidentified cis element downstream of bbd18 may encode a virulence-associated factor critical for infection.
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The primary goal of systems biology is to integrate complex omics data, and data obtained from traditional experimental studies in order to provide a holistic understanding of organismal function. One way of achieving this aim is to generate genome-scale metabolic models (GEMs), which contain information on all metabolites, enzyme-coding genes, and biochemical reactions in a biological system. Drosophila melanogaster GEM has not been reconstructed to date. Constraint-free genome-wide metabolic model of the fruit fly has been reconstructed in our lab, identifying gaps, where no enzyme was identified and metabolites were either only produced or consume. The main focus of the work presented in this thesis was to develop a pipeline for efficient gap filling using metabolomics approaches combined with standard reverse genetics methods, using 5-hydroxyisourate hydrolase (5-HIUH) as an example. 5-HIUH plays a role in urate degradation pathway. Inability to degrade urate can lead to inborn errors of metabolism (IEMs) in humans, including hyperuricemia. Based on sequence analysis Drosophila CG30016 gene was hypothesised to encode 5- HIUH. CG30016 knockout flies were examined to identify Malpighian tubules phenotype, and shortened lifespan might reflect kidney disorders in hyperuricemia in humans. Moreover, LC-MS analysis of mutant tubules revealed that CG30016 is involved in purine metabolism, and specifically urate degradation pathway. However, the exact role of the gene has not been identified, and the complete method for gap filling has not been developed. Nevertheless, thanks to the work presented here, we are a step closer towards the development of a gap-filling pipeline in Drosophila melanogaster GEM. Importantly, the areas that require further optimisation were identified and are the focus of future research. Moreover, LC-MS analysis confirmed that tubules rather than the whole fly were more suitable for metabolomics analysis of purine metabolism. Previously, Dow/Davies lab has generated the most complete tissue-specific transcriptomic atlas for Drosophila – FlyAtlas.org, which provides data on gene expression across multiple tissues of adult fly and larva. FlyAtlas revealed that transcripts of many genes are enriched in specific Drosophila tissues, and that it is possible to deduce the functions of individual tissues within the fly. Based on FlyAtlas data, it has become clear that the fly (like other metazoan species) must be considered as a set of tissues, each 2 with its own distinct transcriptional and functional profile. Moreover, it revealed that for about 30% of the genome, reverse genetic methods (i.e. mutation in an unknown gene followed by observation of phenotype) are only useful if specific tissues are investigated. Based on the FlyAtlas findings, we aimed to build a primary tissue-specific metabolome of the fruit fly, in order to establish whether different Drosophila tissues have different metabolomes and if they correspond to tissue-specific transcriptome of the fruit fly (FlyAtlas.org). Different fly tissues have been dissected and their metabolome elucidated using LC-MS. The results confirmed that tissue metabolomes differ significantly from each other and from the whole fly, and that some of these differences can be correlated to the tissue function. The results illustrate the need to study individual tissues as well as the whole organism. It is clear that some metabolites that play an important role in a given tissue might not be detected in the whole fly sample because their abundance is much lower in comparison to other metabolites present in all tissues, which prevent the detection of the tissue-specific compound.
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Shrimp farming is one of the activities that contribute most to the growth of global aquaculture. However, this business has undergone significant economic losses due to the onset of viral diseases such as Infectious Myonecrosis (IMN). The IMN is already widespread throughout Northeastern Brazil and affects other countries such as Indonesia, Thailand and China. The main symptom of disease is myonecrosis, which consists of necrosis of striated muscles of the abdomen and cephalothorax of shrimp. The IMN is caused by infectious myonecrosis virus (IMNV), a non-enveloped virus which has protrusions along its capsid. The viral genome consists of a single molecule of double-stranded RNA and has two Open Reading Frames (ORFs). The ORF1 encodes the major capsid protein (MCP) and a potential RNA binding protein (RBP). ORF2 encodes a probable RNA-dependent RNA polymerase (RdRp) and classifies IMNV in Totiviridae family. Thus, the objective of this research was study the IMNV complete genome and encoded proteins in order to develop a system differentiate virus isolates based on polymorphisms presence. The phylogenetic relationship among some totivirus was investigated and showed a new group to IMNV within Totiviridae family. Two new genomes were sequenced, analyzed and compared to two other genomes already deposited in GenBank. The new genomes were more similar to each other than those already described. Conserved and variable regions of the genome were identified through similarity graphs and alignments using the four IMNV sequences. This analyze allowed mapping of polymorphic sites and revealed that the most variable region of the genome is in the first half of ORF1, which coincides with the regions that possibly encode the viral protrusion, while the most stable regions of the genome were found in conserved domains of proteins that interact with RNA. Moreover, secondary structures were predicted for all proteins using various softwares and protein structural models were calculated using threading and ab initio modeling approaches. From these analyses was possible to observe that the IMNV proteins have motifs and shapes similar to proteins of other totiviruses and new possible protein functions have been proposed. The genome and proteins study was essential for development of a PCR-based detection system able to discriminate the four IMNV isolates based on the presence of polymorphic sites
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International audience
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Les gènes, qui servent à encoder les fonctions biologiques des êtres vivants, forment l'unité moléculaire de base de l'hérédité. Afin d'expliquer la diversité des espèces que l'on peut observer aujourd'hui, il est essentiel de comprendre comment les gènes évoluent. Pour ce faire, on doit recréer le passé en inférant leur phylogénie, c'est-à-dire un arbre de gènes qui représente les liens de parenté des régions codantes des vivants. Les méthodes classiques d'inférence phylogénétique ont été élaborées principalement pour construire des arbres d'espèces et ne se basent que sur les séquences d'ADN. Les gènes sont toutefois riches en information, et on commence à peine à voir apparaître des méthodes de reconstruction qui utilisent leurs propriétés spécifiques. Notamment, l'histoire d'une famille de gènes en terme de duplications et de pertes, obtenue par la réconciliation d'un arbre de gènes avec un arbre d'espèces, peut nous permettre de détecter des faiblesses au sein d'un arbre et de l'améliorer. Dans cette thèse, la réconciliation est appliquée à la construction et la correction d'arbres de gènes sous trois angles différents: 1) Nous abordons la problématique de résoudre un arbre de gènes non-binaire. En particulier, nous présentons un algorithme en temps linéaire qui résout une polytomie en se basant sur la réconciliation. 2) Nous proposons une nouvelle approche de correction d'arbres de gènes par les relations d'orthologie et paralogie. Des algorithmes en temps polynomial sont présentés pour les problèmes suivants: corriger un arbre de gènes afin qu'il contienne un ensemble d'orthologues donné, et valider un ensemble de relations partielles d'orthologie et paralogie. 3) Nous montrons comment la réconciliation peut servir à "combiner'' plusieurs arbres de gènes. Plus précisément, nous étudions le problème de choisir un superarbre de gènes selon son coût de réconciliation.
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Spiking neural networks - networks that encode information in the timing of spikes - are arising as a new approach in the artificial neural networks paradigm, emergent from cognitive science. One of these new models is the pulsed neural network with radial basis function, a network able to store information in the axonal propagation delay of neurons. Learning algorithms have been proposed to this model looking for mapping input pulses into output pulses. Recently, a new method was proposed to encode constant data into a temporal sequence of spikes, stimulating deeper studies in order to establish abilities and frontiers of this new approach. However, a well known problem of this kind of network is the high number of free parameters - more that 15 - to be properly configured or tuned in order to allow network convergence. This work presents for the first time a new learning function for this network training that allow the automatic configuration of one of the key network parameters: the synaptic weight decreasing factor.
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Motion planning, or trajectory planning, commonly refers to a process of converting high-level task specifications into low-level control commands that can be executed on the system of interest. For different applications, the system will be different. It can be an autonomous vehicle, an Unmanned Aerial Vehicle(UAV), a humanoid robot, or an industrial robotic arm. As human machine interaction is essential in many of these systems, safety is fundamental and crucial. Many of the applications also involve performing a task in an optimal manner within a given time constraint. Therefore, in this thesis, we focus on two aspects of the motion planning problem. One is the verification and synthesis of the safe controls for autonomous ground and air vehicles in collision avoidance scenarios. The other part focuses on the high-level planning for the autonomous vehicles with the timed temporal constraints. In the first aspect of our work, we first propose a verification method to prove the safety and robustness of a path planner and the path following controls based on reachable sets. We demonstrate the method on quadrotor and automobile applications. Secondly, we propose a reachable set based collision avoidance algorithm for UAVs. Instead of the traditional approaches of collision avoidance between trajectories, we propose a collision avoidance scheme based on reachable sets and tubes. We then formulate the problem as a convex optimization problem seeking control set design for the aircraft to avoid collision. We apply our approach to collision avoidance scenarios of quadrotors and fixed-wing aircraft. In the second aspect of our work, we address the high level planning problems with timed temporal logic constraints. Firstly, we present an optimization based method for path planning of a mobile robot subject to timed temporal constraints, in a dynamic environment. Temporal logic (TL) can address very complex task specifications such as safety, coverage, motion sequencing etc. We use metric temporal logic (MTL) to encode the task specifications with timing constraints. We then translate the MTL formulae into mixed integer linear constraints and solve the associated optimization problem using a mixed integer linear program solver. We have applied our approach on several case studies in complex dynamical environments subjected to timed temporal specifications. Secondly, we also present a timed automaton based method for planning under the given timed temporal logic specifications. We use metric interval temporal logic (MITL), a member of the MTL family, to represent the task specification, and provide a constructive way to generate a timed automaton and methods to look for accepting runs on the automaton to find an optimal motion (or path) sequence for the robot to complete the task.
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The evolution of CRISPR–cas loci, which encode adaptive immune systems in archaea and bacteria, involves rapid changes, in particular numerous rearrangements of the locus architecture and horizontal transfer of complete loci or individual modules. These dynamics complicate straightforward phylogenetic classification, but here we present an approach combining the analysis of signature protein families and features of the architecture of cas loci that unambiguously partitions most CRISPR–cas loci into distinct classes, types and subtypes. The new classification retains the overall structure of the previous version but is expanded to now encompass two classes, five types and 16 subtypes. The relative stability of the classification suggests that the most prevalent variants of CRISPR–Cas systems are already known. However, the existence of rare, currently unclassifiable variants implies that additional types and subtypes remain to be characterized.
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Nous avons investigué la relation entre les polymorphismes de nucléotides simples (SNPs) chez trois gènes/loci candidats : DARC, CXCL2 et le loci ORMDL3-GSDMA-CSF3 situés sur le chromosome 17q21 et les complications neutropéniques et infectieuses qui en résultent durant la chimiothérapie chez les patients atteints de la leucémie lymphoblastique aigue. Ces loci codent pour certaines composantes du système immunitaire altérant la concentration de chémokines et leur distribution (DARC), stimulant le relâchement et la migration des neutophiles de la moelle épinière (CXCL2) et régulant la prolifération et la survie des granulocytes (G-CSF). Il est possible que des polymorphismes dans ces loci lorsqu’associés à de la chimiothérapie puissent mettre des individus suceptibles à un risque plus élevé de complication reliées à la chimiothérapie. Une sélection des marqueurs SNPs dans ces gènes ont été génotypés chez des enfants traités au CHU Ste-Justine pour une ALL entre 1989 et 2005. Après correction pour tests multiples, un polymorphisme DARC rs3027012 situé dans le 5’UTR a été associé à un compte phagocytaire peu élevé (APC<500 et <1000 cellules/µL, p=0.001 and p=0.0005, respectivement) ainsi qu’une hospitalisation due à une neutropénie (p=0.007) ou due à une infection et/ou neutropénie (p=0.007). Un effet protecteur a été identifié pour la mutation non sense Gly42Asp variant rs12075 (p=0.006). Des polymorphismes sur le chromosome 17q2 étaient associés à une hospitalisation due à une infection (rs3859192, p= 0.004) et à une neutropénie (rs17609240, p=0.006) L’infection était aussi modulée par CXCL2 (rs16850408, p=0.008) Cette étude identifie pour la première fois que les loci modulant le décompte des leucocytes et des neutrophiles pourraient jouer un rôle dans de déclenchement de complications dues à la chimiothérapie et pourraient ainsi servir de marqueurs pour un ajustement et un suivi du traitement.
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Le présent travail consiste à proposer un modèle de représentation des notions théoriques et pratiques de la terminologie et de leurs relations sous forme de thésaurus. Selon la norme ISO 25964-1, 2011, « un thésaurus est un vocabulaire contrôlé et structuré dans lequel les concepts sont représentés par des termes, ayant été organisés afin de rendre explicites les relations entre les concepts (…) ». Notre objectif est de créer un outil pédagogique à la suite d’une réflexion théorique englobant différentes perspectives notionnelles au sein de cette discipline. Les enjeux soulevés par la classification des concepts de certains champs de savoir (notamment ceux donnant lieu à différentes perspectives) n’ont pas été approfondis suffisamment dans la littérature de la terminologie, ni dans celle portant sur les thésaurus. Comment décrire des concepts qui sont sujets à des dissensions théoriques entre les différentes écoles de pensée? Comment classer les différentes relations entretenues par les concepts théoriques et les applications pratiques d’une discipline? À ces questions s’ajoute celle de la prise en compte de ces difficultés dans un thésaurus. Nous commençons par délimiter et organiser les concepts saillants du domaine. Ensuite, à l’aide d’un corpus comprenant des publications associées à différentes approches de la terminologie, nous étudions les réalisations linguistiques de ces concepts et leurs relations en contexte, dans le but de les décrire, de les classer et de les définir. Puis, nous faisons l’encodage de ces données à l’aide d’un logiciel de gestion de thésaurus, en respectant les normes ISO applicables. La dernière étape consiste à définir la visualisation de ces données afin de la rendre conviviale et compréhensible. Enfin, nous présentons les caractéristiques fondamentales du Thésaurus de la terminologie. Nous avons analysé et représenté un échantillon de 45 concepts et leurs termes reliés. Les différents phénomènes associés à ces descripteurs comme la multidimensionalité, la variation conceptuelle et la variation dénominative sont aussi représentés dans notre thésaurus.
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The electromagnetic form factors are the most fundamental observables that encode information about the internal structure of the nucleon. The electric ($G_{E}$) and the magnetic ($G_{M}$) form factors contain information about the spatial distribution of the charge and magnetization inside the nucleon. A significant discrepancy exists between the Rosenbluth and the polarization transfer measurements of the electromagnetic form factors of the proton. One possible explanation for the discrepancy is the contributions of two-photon exchange (TPE) effects. Theoretical calculations estimating the magnitude of the TPE effect are highly model dependent, and limited experimental evidence for such effects exists. Experimentally, the TPE effect can be measured by comparing the ratio of positron-proton elastic scattering cross section to that of the electron-proton $\large(R = \frac{\sigma (e^{+}p)}{\sigma (e^{-}p)}\large)$. The ratio $R$ was measured over a wide range of kinematics, utilizing a 5.6 GeV primary electron beam produced by the Continuous Electron Beam Accelerator Facility (CEBAF) at Jefferson Lab. This dissertation explored dependence of $R$ on kinematic variables such as squared four-momentum transfer ($Q^{2}$) and the virtual photon polarization parameter ($\varepsilon$). A mixed electron-positron beam was produced from the primary electron beam in experimental Hall B. The mixed beam was scattered from a liquid hydrogen (LH$_{2}$) target. Both the scattered lepton and the recoil proton were detected by the CEBAF Large Acceptance Spectrometer (CLAS). The elastic events were then identified by using elastic scattering kinematics. This work extracted the $Q^{2}$ dependence of $R$ at high $\varepsilon$ ($\varepsilon > $ 0.8) and the $\varepsilon$ dependence of $R$ at $\langle Q^{2} \rangle \approx 0.85$ GeV$^{2}$. In these kinematics, our data confirm the validity of the hadronic calculations of the TPE effect by Blunden, Melnitchouk, and Tjon. This hadronic TPE effect, with additional corrections contributed by higher excitations of the intermediate state nucleon, largely reconciles the Rosenbluth and the polarization transfer measurements of the electromagnetic form factors.
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The present work describes the molecular characterization of five circular plasmids found in the human clinical strain Lactococcus garvieae 21881. The plasmids were designated pGL1-pGL5, with molecular sizes of 4,536 bp, 4,572 bp, 12,948 bp, 14,006 bp and 68,798 bp, respectively. Based on detailed sequence analysis, some of these plasmids appear to be mosaics composed of DNA obtained by modular exchange between different species of lactic acid bacteria. Based on sequence data and the derived presence of certain genes and proteins, the plasmid pGL2 appears to replicate via a rolling-circle mechanism, while the other four plasmids appear to belong to the group of lactococcal theta-type replicons. The plasmids pGL1, pGL2 and pGL5 encode putative proteins related with bacteriocin synthesis and bacteriocin secretion and immunity. The plasmid pGL5 harbors genes (txn, orf5 and orf25) encoding proteins that could be considered putative virulence factors. The gene txn encodes a protein with an enzymatic domain corresponding to the family actin-ADP-ribosyltransferases toxins, which are known to play a key role in pathogenesis of a variety of bacterial pathogens. The genes orf5 and orf25 encode two putative surface proteins containing the cell wall-sorting motif LPXTG, with mucin-binding and collagen-binding protein domains, respectively. These proteins could be involved in the adherence of L. garvieae to mucus from the intestine, facilitating further interaction with intestinal epithelial cells and to collagenous tissues such as the collagen-rich heart valves. To our knowledge, this is the first report on the characterization of plasmids in a human clinical strain of this pathogen.
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Nous avons investigué la relation entre les polymorphismes de nucléotides simples (SNPs) chez trois gènes/loci candidats : DARC, CXCL2 et le loci ORMDL3-GSDMA-CSF3 situés sur le chromosome 17q21 et les complications neutropéniques et infectieuses qui en résultent durant la chimiothérapie chez les patients atteints de la leucémie lymphoblastique aigue. Ces loci codent pour certaines composantes du système immunitaire altérant la concentration de chémokines et leur distribution (DARC), stimulant le relâchement et la migration des neutophiles de la moelle épinière (CXCL2) et régulant la prolifération et la survie des granulocytes (G-CSF). Il est possible que des polymorphismes dans ces loci lorsqu’associés à de la chimiothérapie puissent mettre des individus suceptibles à un risque plus élevé de complication reliées à la chimiothérapie. Une sélection des marqueurs SNPs dans ces gènes ont été génotypés chez des enfants traités au CHU Ste-Justine pour une ALL entre 1989 et 2005. Après correction pour tests multiples, un polymorphisme DARC rs3027012 situé dans le 5’UTR a été associé à un compte phagocytaire peu élevé (APC<500 et <1000 cellules/µL, p=0.001 and p=0.0005, respectivement) ainsi qu’une hospitalisation due à une neutropénie (p=0.007) ou due à une infection et/ou neutropénie (p=0.007). Un effet protecteur a été identifié pour la mutation non sense Gly42Asp variant rs12075 (p=0.006). Des polymorphismes sur le chromosome 17q2 étaient associés à une hospitalisation due à une infection (rs3859192, p= 0.004) et à une neutropénie (rs17609240, p=0.006) L’infection était aussi modulée par CXCL2 (rs16850408, p=0.008) Cette étude identifie pour la première fois que les loci modulant le décompte des leucocytes et des neutrophiles pourraient jouer un rôle dans de déclenchement de complications dues à la chimiothérapie et pourraient ainsi servir de marqueurs pour un ajustement et un suivi du traitement.
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Le présent travail consiste à proposer un modèle de représentation des notions théoriques et pratiques de la terminologie et de leurs relations sous forme de thésaurus. Selon la norme ISO 25964-1, 2011, « un thésaurus est un vocabulaire contrôlé et structuré dans lequel les concepts sont représentés par des termes, ayant été organisés afin de rendre explicites les relations entre les concepts (…) ». Notre objectif est de créer un outil pédagogique à la suite d’une réflexion théorique englobant différentes perspectives notionnelles au sein de cette discipline. Les enjeux soulevés par la classification des concepts de certains champs de savoir (notamment ceux donnant lieu à différentes perspectives) n’ont pas été approfondis suffisamment dans la littérature de la terminologie, ni dans celle portant sur les thésaurus. Comment décrire des concepts qui sont sujets à des dissensions théoriques entre les différentes écoles de pensée? Comment classer les différentes relations entretenues par les concepts théoriques et les applications pratiques d’une discipline? À ces questions s’ajoute celle de la prise en compte de ces difficultés dans un thésaurus. Nous commençons par délimiter et organiser les concepts saillants du domaine. Ensuite, à l’aide d’un corpus comprenant des publications associées à différentes approches de la terminologie, nous étudions les réalisations linguistiques de ces concepts et leurs relations en contexte, dans le but de les décrire, de les classer et de les définir. Puis, nous faisons l’encodage de ces données à l’aide d’un logiciel de gestion de thésaurus, en respectant les normes ISO applicables. La dernière étape consiste à définir la visualisation de ces données afin de la rendre conviviale et compréhensible. Enfin, nous présentons les caractéristiques fondamentales du Thésaurus de la terminologie. Nous avons analysé et représenté un échantillon de 45 concepts et leurs termes reliés. Les différents phénomènes associés à ces descripteurs comme la multidimensionalité, la variation conceptuelle et la variation dénominative sont aussi représentés dans notre thésaurus.
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Image and video compression play a major role in the world today, allowing the storage and transmission of large multimedia content volumes. However, the processing of this information requires high computational resources, hence the improvement of the computational performance of these compression algorithms is very important. The Multidimensional Multiscale Parser (MMP) is a pattern-matching-based compression algorithm for multimedia contents, namely images, achieving high compression ratios, maintaining good image quality, Rodrigues et al. [2008]. However, in comparison with other existing algorithms, this algorithm takes some time to execute. Therefore, two parallel implementations for GPUs were proposed by Ribeiro [2016] and Silva [2015] in CUDA and OpenCL-GPU, respectively. In this dissertation, to complement the referred work, we propose two parallel versions that run the MMP algorithm in CPU: one resorting to OpenMP and another that converts the existing OpenCL-GPU into OpenCL-CPU. The proposed solutions are able to improve the computational performance of MMP by 3 and 2:7 , respectively. The High Efficiency Video Coding (HEVC/H.265) is the most recent standard for compression of image and video. Its impressive compression performance, makes it a target for many adaptations, particularly for holoscopic image/video processing (or light field). Some of the proposed modifications to encode this new multimedia content are based on geometry-based disparity compensations (SS), developed by Conti et al. [2014], and a Geometric Transformations (GT) module, proposed by Monteiro et al. [2015]. These compression algorithms for holoscopic images based on HEVC present an implementation of specific search for similar micro-images that is more efficient than the one performed by HEVC, but its implementation is considerably slower than HEVC. In order to enable better execution times, we choose to use the OpenCL API as the GPU enabling language in order to increase the module performance. With its most costly setting, we are able to reduce the GT module execution time from 6.9 days to less then 4 hours, effectively attaining a speedup of 45 .