Thesis (Ph. D.)--University of Oklahoma. / Includes bibliographical references.

Οι μηνισκικές ρήξεις διακρίνονται σε τραυματικές και εκφυλιστικές. Κλινικά δεδομένα υποδηλώνουν ότι η εκφύλιση των μηνίσκων συσχετίζεται με την οστεοαρθρίτιδα του γόνατος. Παρ’ όλα αυτά, τα μοριακά γεγονότα που καθορίζουν την παθογένεια της εκφύλισης των μηνίσκων σε ανθρώπους παραμένουν αδιευκρίνιστα. Στη μελέτη εξετάστηκε ανοσοϊστοχημικά η έκφραση της p38 MAPKινάσης, της ενεργού φωσφορυλιομένης μορφής της p-p38, του στόχου NF-kB (με τα διμερή p50-p65) καθώς και της COX-2 σε μηνισκικές ρήξεις, και διερευνήθηκε η συμμετοχή τους στην ανάπτυξη εκφύλισης. Τα ευρήματα απέδειξαν αυξημένη έκφραση του άξονα p38-NF-kB και της COX-2 στον αποδιοργανωμένο και εκφυλισμένο ινοχόνδρινο μηνισκικό ιστό, υποδεικνύοντας ένα ρόλο των μορίων αυτών στην παθοβιοχημεία της εκφύλισης και της επακόλουθης ρήξης. Η μελέτη είχε σκοπό να διερευνήσει και να χαρακτηρίσει την έκφραση και την ενεργοποίηση του σηματοδοτικού μονοπατιού της p38 MAPK-NF-kB και της COX-2 στα ινοχονδροκύτταρα των ανθρώπινων μηνίσκων με ρήξη. Επιπρόσθετα συσχετίσαμε τα επίπεδα έκφρασης των πρωτεϊνών αυτών με παθολογοανατομικές και κλινικές παραμέτρους, όπως η ύπαρξη εκφύλισης και η συνύπαρξη κλινικά εξακριβωμένης ΟΑ. Χρησιμοποιήθηκαν 57 ανθρώπινοι μηνίσκοι. 43 (75,4%) άνδρες και 14 (24,6%) γυναίκες, με μέσο όρο ηλικίας 32,6 έτη, με διάρκεια πόνου 17,36 μήνες. Σε 39 ασθενείς (68,4%) η ρήξη αποδόθηκε σε τραύμα και σε 18 (31,6%) σε προϋπάρχουσα κλινικά διαγνωσμένη ΟΑ. Η ιστοπαθολογοανατομική διευκρίνιση της μηνισκικής εκφύλισης βασίστηκε σε καθιερωμένα μικροσκοπικά κριτήρια. Εκφύλιση παρατηρήθηκε σε 34 μηνίσκους (59,4%). Χρησιμοποιήθηκαν τα αντισώματα, anti p38, anti p-p38 (μονοκλωνικά αντισώματα έναντι της ενεργοποιημένης μορφής), anti NF-kB, p50 πολυκλωνική, anti NFkBp65 πολυκλωνική και antiCOX-2. Η ένταση της χρώσης και η αναλογία των ανοσοθετικών ινοχονδροκυττάρων εκτιμήθηκε μικροσκοπικά και βαθμολογήθηκε σε κλίμακα 0-3 (0= χωρίς ανοσοδραστικότητα, 1= ήπια, 2= μέτρια, 3= ισχυρή). Στατιστική ανάλυση έγινε με τη δοκιμασία Mann-Whitney και η ισχύς της συσχέτισης των μεταβλητών με το Kendall’s T test, χρησιμοποιώντας το SPSS. 1.Η έκφραση της p38 ήταν στατιστικά σημαντικά υψηλότερη στους εκφυλισμένους συγκριτικά με μη εκφυλισμένους μηνίσκους. Μηνίσκοι σε ασθενείς με προϋπάρχουσα ΟΑ έδειξαν επίσης στατιστικά σημαντικά αυξημένη p38 σε σύγκριση με αυτούς με μη προϋπάρχουσα ΟΑ. 2.Η p-p38 είχε σημαντικά αυξημένη έκφραση σε εκφυλισμένους μηνίσκους έναντι μη εκφυλισμένων και σε ραγέντες μηνίσκους με ΟΑ σε σχέση με μη ΟΑ. 3.Οι υπομονάδες p50 και p65 ως τροποποιητές του άξονα p38-NFkB έδειξαν σημαντικά υψηλότερα επίπεδα στην εκφύλιση και την οστεοαρθρίτιδα. 4.Τα επίπεδα της COX-2 ήταν σημαντικά διαφορετικά μεταξύ εκφυλισμένων και μη εκφυλισμένων και σε οστεοαρθριτικές και μη αρθρώσεις, και συσχετίζονται απόλυτα με τη διακύμανση των προηγούμενων βημάτων του άξονα. 5.Η στατιστική ανάλυση αποκάλυψε σημαντική και θετική συσχέτιση μεταξύ COX-2 και p38-NF-kB και παράλληλη διακύμανσή της. / Meniscal tears are attributed to either trauma or degeneration processes. Clinical data suggest that meniscal degeneration (MD) is associated with knee osteoarthritis; however, the molecular events underpinning the pathogenesis of MD in humans remain elusive. Here we immunohistochemically examined the expression of p38 MAPK, its phosphorylated activated form p-p38, its target NF-kB (p50-p65 dimer), and COX-2 in ruptured menisci and investigated their involvement in MD development. Our findings demonstrate increased expression of the p38-NF-kB axis elements and COX-2 in disintegrated fibrocartilage suggesting a role of these molecules in the pathobiochemistry of MD and consequential rupture. We undertook this study to explore and characterize the expression and/or activation profile of the p38 MAPK-NF-kB signaling path away constituents and COX-2 in the fibrochondrocytes of human torn menisci. Furthermore we correlated the expression levels of the examined proteins with pathologic and clinical parameters, such as the presence of fibrocartilaginus degeneration and the coexistence of clinically identified OA. 57 human menisci were used for this study. Among the patients 43 (75,4%) were male and 14 (24,6%) female with mean age 32,6 years, with pain duration 17,36 months. In 39 patients (68,4%) meniscal tearing was attributed to trauma and in 18 (31,6%) to a background of clinically diagnosed OA. The histopathologic identification of meniscal degeneration (MD) was based on established microscopy criteria. MD was observed in 34 (59,4%) of the menisci. The following available antibodies were employed (anti-p 38), anti p-p38 (activated form monoclonal), anti NF-kB, p50 polyclonal, anti NFkBp65 polyclonal and anti COX-2. Strain intensity and proportion of immunopositive fibrochondrocytes was assessed by light microscopy and graded on a scale 0-3 (0= no immunoreachivity, 1= mild, 2= moderate, 3= strong). Statistical analysis was made with Mann-Whitney tests and the strength of association between the variables by Kendall’s T test, using SPSS for Windows. 1.Expression of p-38 was significantly higher in degenerated compared with non degenerated menisci. Menisci from patients with preexisting OA showed significantly increased p38 expression levels compared to those with no preexisting OA. 2.p-p38 expression was considerably elevated in degenerated compared with non degenerated and in OA compared with non OA ruptured menisci. 3.The downstream effectors of p38 NF-kB subunits p50 and p65, exhibited significantly higher levels in degenerated and OA fibrocartilage. 4.COX-2 levels were significantly different between degenerated and non degenerated menisci, as well as between OA and non OA joints. 5.Statistical analysis revealed significant and positive correlation between COX-2 and p-38 and NF-kB.

Orientador: Frederico Ozanam Papa / Resumo: O presente estudo propõe avaliar o efeito do tratamento oral com pentoxifilina sobre a qualidade seminal, morfometria, histologia e expressão gênica testicular de garanhões submetidos ao estresse térmico escrotal. Além disso, objetiva testar a eficiência de novo método de aquecimento escrotal na espécie equina. Para isso 14 garanhões foram divididos em três grupos: Controle (CT, n=4), Degenerado (DG, n=5) e Degenerado Tratado (PTX, n=5). O insulto térmico escrotal foi realizada utilizando uma bolsa térmica acoplada a uma fonte de ar aquecido a 50º C, durante uma hora no início da manhã e uma hora no final da tarde, no D-1 e D0. Um dia após o insulto (D1), o tratamento com 17 mg/kg pentoxifilina oral, a cada 12 h, foi iniciado e conduzido por 30 dias. Os animais foram coletados duas vezes por semana do D-24 ao D60 e o sêmen avaliado quanto a cinética, morfologia espermática, integridade de membrana plasmática, geração do ânion superóxido intracelular e mitocondrial, índice de peroxidação lipídica e índice de caspases ativadas 3 e 7. No D30 e D60 biópsias testiculares foram realizadas e as amostras destinadas a histopatologia, e ao RT-qPCR, quanto a resposta a apoptose, choque térmico e estresse oxidativo. As medidas testiculares de comprimento, altura e largura foram mensuradas utilizando paquímetro, e o volume testicular relativo foi calculado, uma vez por semana do D-5 ao D60. Dentre os resultados obtidos, o método de estresse térmico escrotal utilizado se mostrou eficiente ... (Resumo completo, clicar acesso eletrônico abaixo) / Doutor

The retina is a thin layer of tissue lining the back of the eye and is primarily responsible for sight in vertebrates. The neural retina has a distinct layered structure with three dense nuclear layers, separated by plexiform layers comprising of axons and dendrites, and a layer of photoreceptor segments. The retinal and choroidal vasculatures nourish the retina from either side, with an avascular layer comprised largely of photoreceptor cells. Diseases that directly affect the neural retina like retinal degeneration as well as those of vascular origin like diabetic retinopathy can lead to partial or total blindness. Early detection of these diseases can potentially pave the way for a timely intervention and improve patient prognosis. Current techniques of retinal imaging rely mainly on optical techniques, which have limited depth resolution and depend mainly on the clarity of visual pathway. Magnetic resonance imaging is a versatile tool that has long been used for anatomical and functional imaging in humans and animals, and can potentially be used for retinal imaging without the limitations of optical methods. The work reported in this thesis involves the development of high resolution magnetic resonance imaging techniques for anatomical and functional imaging of the retina in rats. The rats were anesthetized using isoflurane, mechanically ventilated and paralyzed using pancuronium bromide to reduce eye motion during retinal MRI. The retina was imaged using a small, single-turn surface coil placed directly over the eye. The several physiological parameters, like rectal temperature, fraction of inspired oxygen, end-tidal CO2, were continuously monitored in all rats. MRI parameters like T1, T2, and the apparent diffusion coefficient of water molecules were determined from the rat retina at high spatial resolution and found to be similar to those obtained from the brain at the same field strength. High-resolution MRI of the retina detected the three layers in wild-type rats, which were identified as the retinal vasculature, the avascular layer and the choroidal vasculature. Anatomical MRI performed 24 hours post intravitreal injection of MnCl2, an MRI contrast agent, revealed seven distinct layers within the retina. These layers were identified as the various nuclear and plexiform layers, the photoreceptor segment layer and the choroidal vasculature using Mn54Cl2emulsion autoradiography. Blood-oxygenlevel dependent (BOLD) functional MRI (fMRI) revealed layer-specific vascular responses to hyperoxic and hypercapnic challenges. Relative blood volume of the retina calculated by using microcrystalline iron oxide nano-colloid, an intravascular contrast agent, revealed a superfluous choroidal vasculature. Fractional changes to blood volume during systemic challenges revealed a higher degree of autoregulation in the retinal vasculature compared to the choroidal vasculature, corroborating the BOLD fMRI data. Finally, the retinal MRI techniques developed were applied to detect structural and vascular changes in a rat model of retinal dystrophy. We conclude that retinal MRI is a powerful investigative tool to resolve layerspecific structure and function in the retina and to probe for changes in retinal diseases. We expect the anatomical and functional retinal MRI techniques developed herein to contribute towards the early detection of diseases and longitudinal evaluation of treatment options without interference from overlying tissue or opacity of the visual pathway.

No description available.

Progressive rod-cone degeneration (PRCD) is the late form of progressive retinal atrophy (PRA). It is an autosomal recessive hereditary retinal defect. This disease in dogs is consistent with one form of retinitis pigmentosa (RP) in humans. Phenotypic manifestations are identical and it is known to be an identical causal mutation. A study of this defect in dogs could also explain a lot in human medicine. The gene for PRCD was mapped in the region of centromer of the canine chromosome 9 (CFA9). In this thesis, genotyping of 120 dogs of different breeds and age was performed. Most represented a breed of English Cocker Spaniel which is predisposed to the disease. Analysis PRA-PRCD was performed by molecular genetic methods PCR-RFLP and the horizontal agarose electrophoresis. Genotypes were determined on the basis of different fragment lengths. The normal allele was 396 bp in length and the mutated allele had a length of 116 bp. Presence of mutated allele was only detected in 25 heterozygotes carriers which were usually breeds with this predisposition. Frequency of the mutated allele was 10.4 %. In the selected population 20.8 % of heterozygotes were represented. The results of the study show approximately one fifth of the tested dogs are heterozygous carriers. Findings of other studies confirm there are generally more heterozygotes than homozygotes in which the disease is manifested during life. However, if this fact is not clearly taken in consideration, the number of sick dogs can rapidly increase during short period of time. In the future, it would be appropriate to adopt measures which would definitely eliminate the occurrence of the mutated allele. These measures could include genetic tests that reliably reveal hidden carriers (heterozygotes) in predisposing breeds. Heterozygotes may increase the representation of this allele in the population. This leads to an increase in the number of diseased animals.

Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder characterized by widespread loss of lower motor neurons from the spinal cord. Lower motor neuron degeneration leads to a progressive decline in motor development, manifesting as muscle atrophy and weakness. It is now well characterised that ubiquitin homeostasis is altered in SMA and that reduction of the ubiquitin-like modifier-activating enzyme 1 (UBA1) is central to this disruption. UBA1 is responsible for activating ubiquitin as the first step in the ubiquitin conjugation process, marking unwanted proteins for degradation by the proteasome. While it is known that therapies targeting UBA1 rescue neuromuscular phenotypes in SMA models, the mechanism by which UBA1 mediates neurodegeneration is unclear. In fact, very little is known about the function of UBA1 beyond its canonical role in the ubiquitin proteasome system. To better understand the role of UBA1 in motor neuron degeneration, a robust set of antibodies for both in vivo and in vitro work to study UBA1 have been identified. This enabled a novel characterisation of UBA1 distribution throughout disease progression in SMA spinal motor neurons to be performed, revealing that UBA1 reduction is an important pre-symptomatic molecular feature of SMA. To identify downstream targets of UBA1 critical for UBA1-mediated degeneration in SMA, label-free proteomics was performed on HEK293 cells after overexpression or knockdown of UBA1. The proteomics data was analysed across multiple platforms, including Biolayout, IPA and DAVID to identify UBA1-dependent pathways and demonstrated that modulation of UBA1 levels lead to disruption of key cellular pathways including translation elongation, nuclear transport, and tRNA synthetases. Validation of target proteins from these UBA1-dependent pathways identified that the tRNA synthetease GARS behaves in a UBA1-dependent manner across a range of model systems in vitro and in vivo. It was then identified that GARS expression is significantly dysregulated across a range of neuronal tissues in a mouse model of SMA. Interestingly, mutations in GARS cause Charcot-Marie-Tooth disease type 2D (CMT2D), an axonal neuropathy, in which a disruption to sensory neuron fate in dorsal root ganglia has recently been identified. In a mouse model of SMA we identified a phenotype consistent with that in the CMT2D mouse model and showed that disruption to sensory neuron fate is reversible and dependent on changes in UBA1 and GARS expression in SMA. In conclusion, modulation of UBA1 levels leads to disruption of key cellular pathways, with dysregulation of tRNA synthetases a prominent feature that is likely to play a role in the pathogenesis of SMA.

La rétinopathie diabétique est la cause principale de cécité dans la population en âge de travailler. Une détection précoce et un traitement adapté permettent de réduire considérablement le risque de perte de vue. Les autorités médicales recommandent un examen annuel pour les patients diabétiques. Plusieurs programmes de dépistage de la rétinopathie diabétique ont été déployés pour appliquer cette recommandation. L'objectif du projet TeleOphta était de détecter automatiquement des examens normaux dans un système de dépistage du diabète, afin de réduire le fardeau des lecteurs, et donc servir plus de patients. Cette thèse propose plusieurs méthodes pour extraire des informations liées à des lésions provoquées par la rétinopathie diabétique dans des images en couleurs du fond d'œil.La détection des exsudats, microanévrismes et hémorragies est discutée en détail. L'un des principaux défis de ce travail est de traiter des images cliniques, acquises avec différents types de caméras de fond d'œil, par des personnes différentes. Par conséquent, l'hétérogénéité de la base de données est élevé. Des nouvelles méthodes de pré-traitement, qui effectuent non seulement des tâches de normalisation et de débruitage, mais aussi de détection de réflexions et d'artefacts optiques, sont proposées. Des méthodes de segmentation des candidats basées sur la morphologie mathématique, et de nouveaux descripteurs de texture et de contexte sont proposées pour la caractérisation des lésions. Un algorithme de forêts aléatoires est utilisé pour choisir les lésions parmi les candidats. Les méthodes proposées utilisent largement des nouvelles méthodes d'analyse des résidus.En outre, trois nouvelles bases de données publiques d'images de la rétine, e-ophtha EX, e-ophtha MA et e-ophtha HM, respectivement conçues pour développer et évaluer les méthodes de détection d' exsudats,de microanévrismes et d'hémorragies, sont proposées dans ce travail. Les images ont été extraites du réseau de télémédecine OPHDIAT pour le dépistage de la rétinopathie diabétique. Des annotations manuelles détaillées des lésions sont fournies avec ces bases de données. Les algorithmes proposés sont évalués sur ces bases.Les méthodes proposées ont été intégrées dans le système TeleOphta , qui est présentée et évaluée sur deux grandes bases de données. Chaque dossier du patient est classé en deux catégories: “Pour avis” ou “Normal". La classification est basée non seulement sur les résultats des méthodes présentées, mais aussi sur les signatures d'image fournies par d'autres partenaires, ainsi que sur l'information médicale du patient, et les données liées à l'acquisition. L'évaluation montre que le système TeleOphta permet de traiter deux fois plus de patients dans un réseau de dépistage, à moyens constants. / Diabetic retinopathy is the main cause of blindness among the middle-aged population. An early detection and adapted treatment considerably reduce the risk of sight loss. Medical authorities recommend an annual examination to diabetic patients. Several diabetic retinopathy screening programs have been deployed to enforce this recommendation. The aim of the TeleOphta project was to automatically detect normal examinations in a diabetic screening system, in order to reduce the burden on readers, and therefore serve more patients. This thesis proposes several methods to extract information linked to diabetic retinopathy lesions from color eye fundus images.The detection of exudates, microaneurysms and hemorrhages is discussed in detail. One of the main challenges of this work is to deal with clinical images, acquired by different types of eye fundus cameras, by different persons. Therefore the data base heterogeneity is high. New pre-processing methods, which perform not only normalization and denoising tasks, but also detect reflections and artifacts in the images, are proposed. Novel candidate segmentation methods based on mathematical morphology, and new textural and contextual features for lesion characterization, are proposed. A random forest algorithm is used to detect lesions among the candidates. The proposed methods make extensive use of new residue analysis methods.Moreover, three new publicly available retinal image databases, e-ophtha EX, e-ophtha MA and e-ophtha HM, respectively designed to develop and evaluate exudate, microaneurysms and hemorrhages detections methods, are proposed in this work. The images are extracted from the OPHDIAT telemedicine network for diabetic retinopathy screening. Manual annotations of the lesions are given in detail in these databases. The proposed algorithms are evaluated on these databases.The proposed methods have been integrated within the TeleOphta system, which is presented and evaluated on two large databases. Each patient record is classified into two categories: “To be referred” or “Normal”. The classification is based not only on the results of the presented methods, but also on image signatures provided by other partners, as well as on medical and acquisition-related information. The evaluation shows that the TeleOphta system can make about 2 times more patients benefit from the diagnosis service.

Parkinson’s disease (PD) is a debilitating movement disorder. The cardinal symptoms of PD are bradykinesia, resting tremors and rigidity. PD is characterized by degeneration of dopaminergic neurons of A9 region, substantia nigra pars compacta (SNpc) and loss of dopaminergic terminals in striatum while the dopaminergic neurons of A10 region, ventral tegmental area (VTA) are relatively protected. Putative mechanisms, such as mitochondrial dysfunction, dysregulation of the ubiquitin proteasome system and increased oxidative stress have been hypothesized to mediate PD pathology. However, precise mechanisms that underlie selective vulnerability of SNpc dopaminergic neurons to degeneration are unknown. The aim of this thesis was to evaluate the pathological mechanisms that may contribute to degeneration of SNpc dopaminergic neurons in PD. Dopaminergic neurons of SNpc are pacemakers and constant calcium entry through L-type calcium channel, Cav1.3 has been reported in these neurons during pacemaking. In addition, these neurons have poor calcium buffering capacity. Together, this leads to dysregulation of calcium homeostasis in the SNpc dopaminergic neurons leading to increased oxidative stress. Gene expression of the full length channel and the variant was investigated in the mouse midbrain and further their presence was verified in mouse SNpc and VTA and also in SNpc and VTA in the MPTP mouse model of PD. Gene expression of Cav1.3 -42 and its variant was also studied in SNpc from autopsy tissue from PD patients and age matched controls. Having studied differential expression of the calcium channels, global changes in gene expression in SNpc from the MPTP mouse model of PD and PD autopsy tissues were next examined. This is the first report of transcriptome profile alterations from SNpc in mouse model and PD tissue performed using RNA-seq. Gene expression profiles were examined from SNpc 1 day post single exposure to MPTP, in which case there is no neuronal death and 14 days after daily MPTP treatment where SNpc has undergone ~50% cell death. Further, RNA- seq was performed to study gene expression alterations in SNpc from human PD patients and age- matched controls. The RNA-seq data was taken through extensive analyses; analysed for differential gene expression, gene-set enrichment analysis, pathway analysis and network analysis. Glutaredoxin 1 (Grx1) is a thiol disulfide oxidoreductase that catalyses the deglutathionylation of proteins and is important for regulation of cellular protein thiol redox homeostasis. Down-regulation of Grx1 has been established to exacerbate neurodegeneration through impairment of cell survival signalling. Previous work from our laboratory has demonstrated that perturbation of protein thiol redox homeostasis through diamide injection into SNpc leads to development of PD pathology and motor deficits. It was therefore investigated if Grx1 down-regulation in vivo, leading to increased glutathionylation and protein thiol oxidation, could result in PD pathology. This work is thus the first study of RNA-seq based transcriptomic profile alterations in SNpc from human PD patients. This work also highlights several differences between mouse model and human PD tissue indicating that the underlying mechanisms of PD pathogenesis differ from mouse to humans in addition to developing a novel model for PD.

Introdução: Pacientes que sobrevivem ao traumatismo crânio-encefálico (TCE) apresentam declínio cognitivo e sinais indiretos de atrofia cerebral maiores que o esperado para a população normal. Dentro do universo das lesões englobadas sob o termo TCE há diferentes tipos de lesões, que podem ser divididas entre focais e difusas. A lesão axonial difusa (LAD), está presente em quase todos os pacientes com TCE moderado e grave. Não há estudos que descrevam longitudinalmente o que ocorre nos exames de imagem após o TCE em um grupo com diagnóstico clínico e radiológico de LAD sem lesões focais significativas. Este estudo tem como objetivo avaliar a carga de lesões da LAD através de uma contagem sistematizada, avaliar a taxa de atrofia de diferentes compartimentos do encéfalo de forma longitudinal, e verificar se o número de lesões mostra correlação com tais taxas de atrofia e ou com testes neuropsicológicos que avaliam desempenho executivo e de memória. Método: Foram selecionados 24 pacientes com diagnóstico clínico-radiológico de LAD e realizados exames de RM nos meses 2 (fase 1), 6 (fase 2) e 12 (fase 3) após o TCE. Nas fases 2 e 3 foi realizada avaliação neuropsicológica. Foi realizada contagem de lesões segundo a Microbleed Anatomical and Rating Scale (MARS). Nos definidos momentos foi realizada avaliação do volume do encéfalo através do software FreeSurfer. Foram avaliados a capacidade executiva através dos testes Trail Making Test (TMT) A e B, e a capacidade de recordação através do teste Hopkins Verbal Learning Test (HLVT) em seus componente de recordação imediata (HVLT-RI), tardia (RVLT-RT) e reconhecimento (HVLT-R). Foi testada correlação da carga lesional com a redução de volume dos compartimentos substância branca (VSB), substância cinzenta cortical (VCC), substância cinzenta subcortical (VCS) volume cerebral total (VCT). Foram ainda realizados testes de correlação da carga lesional total e por sítio anatômico com os testes TMT e HVLT e de correlação do grau de atrofia do VSB, VCC, VCS e VCT com os testes HVLT e TMT. Foram considerados positivos os resultados com p<0,05. Resultados: O VSB foi significativamente diferente entre as fases 2 e 3 e entre as fases 1 e 3, com redução de volume de 4,0% no intervalo total do estudo. O VCT foi significativamente diferente entre as fases 2 e 3 meses e entre as fases 1 e 3, com redução de volume de 1,9% no intervalo total do estudo. O VCC não foi significativamente diferente nas 3 fases. O VCS foi significativamente diferente entre as fases 1 e 2; fases 2 e 3 e entre as fases 1 e 3, com redução de volume de 3,7%. O número médio de lesões pela tabela MARS foi de 128 (DP 95), e mostrou correlação positiva e significativa com a redução do VSB, e não demonstrou correlação com a redução de volume dos demais compartimentos. Houve diferença significativa nos resultados dos testes TMT-A e TMT-B entre as fases 2 e 3, com maior rapidez na execução do teste na fase 3. Houve diferença significativa entre os resultados do teste HVLT-RI as fases 2 e 3, com maior número de palavras recordada na fase 3. Não houve diferença significativa nos resultados dos testes HVLT-RT e HVLT-R nas 2 fases. Houve correlação entre o resultado dos testes TMT-B nas fases 2 e 3 com a redução do VCT e entre os resultados do teste TMT-A na fase 3 com a redução do VSB. Não foi encontrada qualquer correlação entre o número de lesões segundo o sítio anatômico da tabela MARS com o desempenho nos testes TMT-A ou TMT-B. Não foi encontrada correlação entre os testes HVLT-RI, HVLT-RT ou HVLT-R com a redução dos volumes de VCT, VSB ou VCC. Discussão e Conclusões: Houve redução significativa do VCT, VSB e VCC ao longo do intervalo entre as fases 1 e 3 do estudo, e simultaneamente houve melhora no desempenho dos testes executivos TMT-A e TMT-B. Tais achados podem ser interpretados como uma resultante daquilo que modelos animais têm demonstrado na evolução do TCE: existe um processo contínuo no tecido cerebral após o TCE, que inclui o clareamento dos debris celulares irremediavelmente lesados e reparação de parte do tecido neural que sofreu lesões reversíveis no momento do trauma, tudo isso contribuindo para uma melhora no desempenho cognitivo, ao mesmo tempo em que ocorre redução do volume dos compartimentos encefálicos. A avaliação da carga lesional mostrou-se de valor prognóstico, pois manteve correlação com o grau de atrofia do VSB no intervalo do estudo / Introduction: Patients who survive traumatic brain injury (TBI) present cognitive decline and indirect signs of brain atrophy greater than expected for the normal population. Within the universe of injuries encompassed under the term TBI there are different types of injuries, which can be divided between focal and diffuse. Diffuse axonal injury (DAI) is present in almost all patients with moderate and severe TBI. There are no longitudinal studies describing imaging findings after TBI in a group with clinical and radiological diagnosis of DAI without significant focal lesions. This study aims to evaluate the DAI lesion load through a systematic counting approach, to evaluate longitudinally the atrophy rate of various brain compartments and to verify correlations between the lesion load and atrophy rates and their correlation with neuropsychological tests evaluating executive and memory performances. Method: 24 patients with clinical and radiological diagnosis of DAI were selected and they were submitted to MRI scans in 2, 6 and 12 months after TBI, as defined as the phase 1, phase 2, and phase 3 of the study. In phases 2 and 3 neuropsychological assessment was performed. Lesion load was quantified according to Microbleed Anatomical and Rating Scale (MARS). In all the 3 phases brain volume assessment was performed by FreeSurfer software. The executive capacity was evaluated by the Trail Making Test (TMT) A and B, and the memory capacity by the Hopkins Verbal Learning Test (HLVT) in its immediate recall component (HVLT-IR), late recall (RVLT-LR) and recognition (HVLT-R). The lesional load was correlated to the reduction in white matter volume (WMV), cortical gray matter volume (CGV), and subcortical gray matter (SGV) and total brain volume (TBV). Correlation of the total lesion load and anatomical site were correlated to TMT and HVLT tests. It was also performed correlation between degree of atrophy of the WMV, CGV, SGV and TGV with HVLT and TMT tests. Positive results were considered with p < 0.05. Results: The WMV was significantly different between phases 2 and 3 and between phases 1 and 3, with volume reduction of 4.0% in the total study interval. TBV was significantly different between the phases 2 and 3 and between phases 1 and 3, with volume reduction of 1.9% in the total study interval. The CGV was not significantly different in any of the 3 phases. The SGV was significantly different between phases 1 and 2, phases 2 and 3 and between phases 1 and 3, with 3.7% volume reduction in the total study interval. The mean lesion load assessment by MARS was 128 (SD 95) and showed a positive and significant correlation with the reduction in the WMV, and no correlation with the volume reduction of the other evaluated compartments. There were significant differences in the results of the TMT-A and TMT-B tests between phases 2 and 3, with faster execution of the test in phase 3. There were significant differences between the HVLT-IR results phases 2 and 3, with the largest number of words recalled in phase 3. There were no significant differences in the results of HVLT-LR tests and HVLT-R in 2 phases. There were correlations between the result of TMT-B test at phases 2 and 3 to the reduction of the TBV and the results of the TMT at phase 3 to the WMV reduction. There were no correlations between the anatomical site lesion load with the performance in the TMT-A and TMT-B. No correlations were found between HVLT-IR, HVLT-LR or HVLT-R with volume reduction of TBV, WMV or CGM. Discussion and Conclusions: There was a significant volume reduction in TBV, WMV and SGV during the study interval, while there was an improvement the executive tests TMT-A and TMT-B performance. These findings can be interpreted as a result of what animal models have shown the evolution of the ECT: there is a continuous process in the brain tissue after TBI, including clearing irreparably damaged cell debris and repair of the neural tissue components that suffered reversible injuries at the moment of trauma. Those processes contribute to an improvement in cognitive performance, while reduction of the volume of the encephalic compartments occurs at the same time. The lesion evaluation has proven its prognostic value as it showed correlation with the degree of WMV reduction