Thesis (Ph.D)--University of Tennessee Health Science Center, 2007. / Title from title page screen (viewed on July 22, 2008). Research advisor: Terrence L. Geiger, MD, Ph.D. Document formatted into pages (xiv, 149 p. : ill.). Vita. Abstract. Includes bibliographical references (p. 140-149).

Multiple Sclerosis (MS) is an autoimmune disease of the central nervous system (CNS) that is characterized by widespread focal areas of inflammation and demyelination. Although the exact cause of the disease is still not known, myelin-reactive T cells that enter the CNS trigger the disease and lead to the recruitment and activation of macrophages and other immune cells. One set of candidates that could serve to mediate these CNS changes is the family of phospholipase A2 (PLA2) enzymes, which consist of secreted (sPLA2) and cytosolic (cPLA2) forms. These enzymes hydrolyze membrane phospholipids to release free fatty acids (arachidonic acid) that can stimulate complex inflammatory cascades, and lysophospholipids that can induce myelin breakdown and demyelination, the two pathological hallmarks of MS. / For my Ph.D. research I studied the expression and role of different members of the PLA2 family in 'experimental autoimmune encephalomyelitis' (EAE), a widely used animal model of MS. I first generated a relapsing-remitting form of EAE in the C57BL/6 mouse strain that lacks a major form of sPLA2. I showed that cPLA2 is expressed by immune cells in the EAE lesions in the CNS. Furthermore blocking the activity of cPLA2 with a broad-spectrum chemical inhibitor starting at the time of EAE induction reduced the incidence and severity of disease, reduced lesion burden as well as reduced the expression of a number of chemokines and cytokines. Treating mice in the remission phase also prevented further clinical episodes. This showed that some or all members of the cPLA2 family play an important role in the onset and progression of EAE in a strain of mice lacking sPLA2. / I next carried out studies to assess the expression of all 14 members of the sPLA2 and cPLA2 families at the onset, peak and remission stages of EAE in the SJL/J mouse strain that expresses all forms of PLA2. The mRNA expression of only 4 of these PLA2s was increased. These include sPLA2 (groups IIA and V) and cPLA 2 (groups IVA and VIA). The expression of these PLA2s in the CNS was also characterized by double-immunofluorescence. The role of these PLA2s was assessed using selective inhibitors and analysed by monitoring the clinical disability scores, chemokine/cytokine protein arrays, lipomics lipid profiling, and histological analysis. Surprisingly, the sPLA2 inhibitor prevented disease remission and worsened the clinical outcome. This was accompanied by an increase in several pro-inflammatory chemokines. Selective inhibitors of cPLA2 group IVA and the calcium independent foam group VIA (iPLA2) reduced severity of EAE when given starting before onset of disease. The cPLA2 inhibitor treatment was effective only while administered, while iPLA2 inhibitor treatment was effective even after treatment was stopped. Furthermore, only delayed treatment with the iPLA2 inhibitor was effective, suggesting that cPLA2 group IVA only plays a role in the initiation of disease, while iPLA 2 plays a role in both disease onset and progression. These effects were also associated with concomitant reduction in chemokine/cytokine expression, reduction of inflammatory lipid mediators, and increase in protective lipids e.g., omega 3 fatty acids. / This work has allowed us to dissect out the expression and role of different members of the PLA2 family and has revealed the importance of selectively inhibiting some but not others in EAE. These findings may therefore have important implications for the treatment of MS.

No description available.

This thesis presents methods for quantitative MRI analysis of brain injury, repair and degeneration in multiple sclerosis (MS) that provide new insights into disease pathogenesis and evolution. / Demyelinated and inflammatory white-matter lesions are hallmark features of MS. A methodology is described to detect regions of acute white-matter lesions that undergo myelin destruction and repair based on analysis of magnetization transfer ratio (MTR) images. Validation is performed based on histopathology and error is assessed based on same-day scans. To quantify the spatial extent and temporal evolution of myelin destruction and repair, data from a 3-year clinical trial is analyzed using this method. Approximately 20% of acute lesion voxels show some repair over the initial 7 months. In subsequent months, there is little further repair, but some increases in the lesion volume undergoing demyelination. / Although less conspicuous on conventional MRI, there is considerable MS pathology in the brain tissue outside of white-matter lesions. An image-processing methodology was developed to obtain accurate metrics that quantify change over time in whole-brain MTR (associated with changes in myelin-density) and in T2 relaxation time (associated with changes in inflammatory edema). These metrics, in addition to metrics of brain atrophy and axonal integrity, were used to quantify brain injury and degeneration following immunoablation and autologous hematopoietic stem cell transplantation therapy for MS. Pronounced brain volume loss was detected immediately following therapy, associated with decreased myelin density and not resolution of edema. / Post-mortem histopathology has revealed abnormalities in the cortical grey-matter of MS patients that appear to be independent of white-matter lesions. A methodology to quantify neocortical injury and degeneration that yields cross-sectional and longitudinal metrics of cortical thickness and grey-matter/white-matter interface integrity both globally and regionally is presented and validated. MS patients with progressive disability showed greater decreases in cortical metrics compared to MS patients with stable disability. / The quantitative MRI analysis methods presented in this thesis are applicable to MRI data obtained in clinical trials of therapies for MS, have the necessary sensitivity and specificity to assess therapeutic efficacy, and provide new insights into disease pathogenesis and evolution.

Brain atrophy has been used as a marker for disease progression in Multiple Sclerosis (MS). SIENA, an automated tool for measuring brain volume change, was tested to see whether MRI slice thickness and gap presence affect longitudinal atrophy measures. Isotropic global scan-rescan images were used to simulate 3 mm and 5 mm axial slice thicknesses with 1 and 2mm gaps, respectively. SIENA remained accurate and precise with increasing slice thickness and gap presence. Furthermore, symmetric pre-registration was crucial for scans with larger slice-thickness and gaps. / SIENA was used to observe atrophy in children who have experienced a Clinically Isolated Syndrome (CIS) of the type leading to MS (CIS-MS). Brain atrophy was present within the first three months after a CIS event, and then subsided over the rest of the year. If the first acute episode was excluded, there was no significant difference in atrophy rates between the CIS-MS group and the CIS group, and no significant difference between those with T2-weighted brain lesions versus those who had none.

No description available.

No description available.

INTRODUÇÃO: Aproximadamente 20 a 30% dos casos novos de epilepsia se tornam refratários ao tratamento medicamentoso ao longo do tempo e a maioria destes casos está relacionada à esclerose mesial temporal (EMT). Disfunção metabólica tem sido relacionada à EMT por diferentes métodos, como tomografia de emissão de pósitrons, tomografia computadorizada por emissão de fóton único, espectroscopia de próton e fósforo (31P-ERM) por ressonância magnética. A caracterização das alterações metabólicas relacionadas à EMT pode auxiliar na melhor compreensão da epileptogênese, da refratariedade clínica comumente encontrada nestes pacientes e na busca de novas possibilidades terapêuticas. OBJETIVOS: Avaliar pela 31P-ERM o perfil metabólico dos pacientes com EMT e comparar com os dados de controles saudáveis. Verificar a existência de correlação entre o tempo de epilepsia, frequência de crises com os resultados da 31P-ERM nas regiões hipocampais. MÉTODOS: 33 pacientes com EMT unilateral e 31 controles foram estudados pela 31P-ERM, tridimensional em aparelho de 3,0 Tesla. Os voxels selecionados para análise foram os localizados na região hipocampal (RH), região insulonuclear anterior (RINA), região insulonuclear posterior (RINP) e na região frontal (RF). Estes voxels foram considerados ipsilaterais ou contralaterais à EMT, diagnosticada pela ressonância magnética convencional. Os metabólitos estudados foram: fosfodiésteres (PDE), fosfomonoésteres (PME), fosfato inorgânico (Pi), fosfocreatina (PCr) , adenosina trifosfato total (ATPt) compostos pela soma dos ?-, ?- e ?-ATP. Foram ainda avaliadas as razões PCr/ATPt, PCr/?-ATP, PCr/Pi e PME/PDE bem como o pH e magnésio. RESULTADOS: Na RH contralateral, aumento de ?-ATP e redução de PCr/ ?-ATP foram significativos. Na RINA ipsilateral, redução de Pi e PCr/ ?-ATP foram detectados. Na RINA contralateral, houve redução de Pi e aumento de ATPT. Na RINP ipsilateral, foram detectados redução de Pi, PCr/?-ATP e aumento de ?-ATP. Na RF contralateral houve redução de Pi. Não houve diferença entre nenhum dos metabólitos ou razões estudados na RH ipsilateral, RINP contralateral, RF ipsilateral em comparação aos controles. Não houve correlação dos resultados da 31P-ERM com o tempo de epilepsia na RH. Em relação à frequência de crises, houve correlação com ATPT, PME, PDE, PME/PDE na RH ipsilateral e de ?-ATP, ATPT, PCr/ ?-ATP e PCr/ATPT na RH contralateral. Análises adicionais foram feitas com os resultados na RH, separando os pacientes de acordo com a frequência de crises e tempo de epilepsia. No grupo de pacientes com frequência igual ou maior que 10 crises por mês, o PME foi menor na RH ipsilateral enquanto que o ?-ATP foi maior e o PCr/?-ATP menor na RH contralateral. No grupo de pacientes com tempo de epilepsia igual ou maior que 20 anos, o ?-ATP foi maior na RH contralateral. Nos grupos de pacientes com frequência de crises menor que 10 por mês ou tempo de epilepsia menor que 20 anos, nenhuma diferença foi detectada. Dados interpolados nas RH foram avaliados, notando-se redução de PME na RH ipsilateral e tendência a significância de diferenças de ?-ATP e PCr/?-ATP na RH contralateral. CONCLUSÕES: Há alterações energéticas difusas nas regiões não epileptogênicas de pacientes com EMT que podem estar relacionadas à disfunção mitocondrial. Na RH ipsilateral, detectou-se redução de PME na análise de voxels interpolados e no grupo de pacientes com frequência maior que 10 crises por mês, indicando alteração de composição de membrana, provavelmente relacionado à gliose e perda neuronal / INTRODUCTION: Around 20-30% of new cases of epilepsy become drug resistant over time and most of these cases are related to mesial temporal sclerosis (MTS). Metabolic dysfunction has been related to MTS by different methods such as positron emission tomography, single photon emission computed tomography, proton and phosphorus (31P-MRS) magnetic resonance imaging spectroscopy. Characterization of metabolic changes related to MTS can improve the understanding of epileptogenesis and drug resistance commonly found in these patients and in the search of new therapeutic possibilities. OBJETIVES: Evaluate by 31P-MRS the metabolic profile of patients with MTS and compare with data from healthy controls. Check the correlation between time of epilepsy, seizure frequency with the results of 31P-MRS in the hippocampal regions. METHODS: 33 unilateral MTS patients and 31 controls were studied by three-dimensional 31P-MRS in 3.0 Tesla equipment. The voxels selected for analysis were located in the hippocampal region (HR), anterior insulonuclear region (RINA), posterior insulonuclear region (RINP) and frontal region (RF). These voxels were considered ipsilateral or contralateral to MTS, diagnosed by conventional MRI. Analised metabolites were: phosphodiesthers (PDE), phosphomonoesthers (PME), inorganic phosphate (Pi), phosphocreatine (PCr) , total adenosine triphosphate ( ATPt) sum of ?- + ?- + ?-ATP and ?-ATP. We also evaluate PCr/ATPt , PCr/?-ATP, PCr/Pi and PME/PDE ratios as well as pH and magnesium. RESULTS: In the contralateral HR, increase of ?-ATP and decrease of PCr / ?-ATP were significant. In the ipsilateral RINA, reductions of Pi and PCr / ?-ATP were detected. In the contralateral RINA, there were reduction of Pi and increased ATPT. In the ipsilateral RINP, reduction of Pi and PCr / ?-ATP and increased ?-ATP were detected. In the contralateral RF, decreased Pi was found. There was no difference between any of the studied metabolites or ratios in the ipsilateral HR, contralateral RINP, ipsilateral RF compared to controls. There was no correlation between the results of 31P-ERM and time of epilepsy in HR. The following metabolites showed correlation with the seizure frequency in the ipsilateral HR: ATPT, PME, PDE, PME/PDE. The seizure frequency was correlated to ?-ATP, ATPT, PCr/ ?-ATP, PCr/ATPT in the contralateral HR. Aditional analysis was made with the results from HR, dividing the patients according to seizure frequency and time of epilepsy. Patients with 10 or more seizures per month showed reduction of PME in the ipsilateral HR; increase of ?-ATP and decrease of PCr/?- ATP in the contralateral HR. Patients with time of epilepsy of 20 years or more showed increase of ?-ATP in the contralateral HR. Patients with less than 10 seizures per month or time of epilepsy lower than 20 years did not demonstrate any difference. Reduction of PME in the ipsilateral HR and tendency to significant differences in ?-ATP and PCr/?-ATP in the contralateral HR were detected from interpolated data. CONCLUSIONS: There are diffuse energetic changes in non epileptogenic regions of patients with MTS that could be related to mitochondrial dysfunction. In the ipsilateral HR, reduction of PME was found from interpolated data and in the group of patients with 10 or more seizures per month, suggesting changes in membrane composition, probably related to gliosis and neuronal loss

INTRODUÇÃO: Aproximadamente 20 a 30% dos casos novos de epilepsia se tornam refratários ao tratamento medicamentoso ao longo do tempo e a maioria destes casos está relacionada à esclerose mesial temporal (EMT). Disfunção metabólica tem sido relacionada à EMT por diferentes métodos, como tomografia de emissão de pósitrons, tomografia computadorizada por emissão de fóton único, espectroscopia de próton e fósforo (31P-ERM) por ressonância magnética. A caracterização das alterações metabólicas relacionadas à EMT pode auxiliar na melhor compreensão da epileptogênese, da refratariedade clínica comumente encontrada nestes pacientes e na busca de novas possibilidades terapêuticas. OBJETIVOS: Avaliar pela 31P-ERM o perfil metabólico dos pacientes com EMT e comparar com os dados de controles saudáveis. Verificar a existência de correlação entre o tempo de epilepsia, frequência de crises com os resultados da 31P-ERM nas regiões hipocampais. MÉTODOS: 33 pacientes com EMT unilateral e 31 controles foram estudados pela 31P-ERM, tridimensional em aparelho de 3,0 Tesla. Os voxels selecionados para análise foram os localizados na região hipocampal (RH), região insulonuclear anterior (RINA), região insulonuclear posterior (RINP) e na região frontal (RF). Estes voxels foram considerados ipsilaterais ou contralaterais à EMT, diagnosticada pela ressonância magnética convencional. Os metabólitos estudados foram: fosfodiésteres (PDE), fosfomonoésteres (PME), fosfato inorgânico (Pi), fosfocreatina (PCr) , adenosina trifosfato total (ATPt) compostos pela soma dos ?-, ?- e ?-ATP. Foram ainda avaliadas as razões PCr/ATPt, PCr/?-ATP, PCr/Pi e PME/PDE bem como o pH e magnésio. RESULTADOS: Na RH contralateral, aumento de ?-ATP e redução de PCr/ ?-ATP foram significativos. Na RINA ipsilateral, redução de Pi e PCr/ ?-ATP foram detectados. Na RINA contralateral, houve redução de Pi e aumento de ATPT. Na RINP ipsilateral, foram detectados redução de Pi, PCr/?-ATP e aumento de ?-ATP. Na RF contralateral houve redução de Pi. Não houve diferença entre nenhum dos metabólitos ou razões estudados na RH ipsilateral, RINP contralateral, RF ipsilateral em comparação aos controles. Não houve correlação dos resultados da 31P-ERM com o tempo de epilepsia na RH. Em relação à frequência de crises, houve correlação com ATPT, PME, PDE, PME/PDE na RH ipsilateral e de ?-ATP, ATPT, PCr/ ?-ATP e PCr/ATPT na RH contralateral. Análises adicionais foram feitas com os resultados na RH, separando os pacientes de acordo com a frequência de crises e tempo de epilepsia. No grupo de pacientes com frequência igual ou maior que 10 crises por mês, o PME foi menor na RH ipsilateral enquanto que o ?-ATP foi maior e o PCr/?-ATP menor na RH contralateral. No grupo de pacientes com tempo de epilepsia igual ou maior que 20 anos, o ?-ATP foi maior na RH contralateral. Nos grupos de pacientes com frequência de crises menor que 10 por mês ou tempo de epilepsia menor que 20 anos, nenhuma diferença foi detectada. Dados interpolados nas RH foram avaliados, notando-se redução de PME na RH ipsilateral e tendência a significância de diferenças de ?-ATP e PCr/?-ATP na RH contralateral. CONCLUSÕES: Há alterações energéticas difusas nas regiões não epileptogênicas de pacientes com EMT que podem estar relacionadas à disfunção mitocondrial. Na RH ipsilateral, detectou-se redução de PME na análise de voxels interpolados e no grupo de pacientes com frequência maior que 10 crises por mês, indicando alteração de composição de membrana, provavelmente relacionado à gliose e perda neuronal / INTRODUCTION: Around 20-30% of new cases of epilepsy become drug resistant over time and most of these cases are related to mesial temporal sclerosis (MTS). Metabolic dysfunction has been related to MTS by different methods such as positron emission tomography, single photon emission computed tomography, proton and phosphorus (31P-MRS) magnetic resonance imaging spectroscopy. Characterization of metabolic changes related to MTS can improve the understanding of epileptogenesis and drug resistance commonly found in these patients and in the search of new therapeutic possibilities. OBJETIVES: Evaluate by 31P-MRS the metabolic profile of patients with MTS and compare with data from healthy controls. Check the correlation between time of epilepsy, seizure frequency with the results of 31P-MRS in the hippocampal regions. METHODS: 33 unilateral MTS patients and 31 controls were studied by three-dimensional 31P-MRS in 3.0 Tesla equipment. The voxels selected for analysis were located in the hippocampal region (HR), anterior insulonuclear region (RINA), posterior insulonuclear region (RINP) and frontal region (RF). These voxels were considered ipsilateral or contralateral to MTS, diagnosed by conventional MRI. Analised metabolites were: phosphodiesthers (PDE), phosphomonoesthers (PME), inorganic phosphate (Pi), phosphocreatine (PCr) , total adenosine triphosphate ( ATPt) sum of ?- + ?- + ?-ATP and ?-ATP. We also evaluate PCr/ATPt , PCr/?-ATP, PCr/Pi and PME/PDE ratios as well as pH and magnesium. RESULTS: In the contralateral HR, increase of ?-ATP and decrease of PCr / ?-ATP were significant. In the ipsilateral RINA, reductions of Pi and PCr / ?-ATP were detected. In the contralateral RINA, there were reduction of Pi and increased ATPT. In the ipsilateral RINP, reduction of Pi and PCr / ?-ATP and increased ?-ATP were detected. In the contralateral RF, decreased Pi was found. There was no difference between any of the studied metabolites or ratios in the ipsilateral HR, contralateral RINP, ipsilateral RF compared to controls. There was no correlation between the results of 31P-ERM and time of epilepsy in HR. The following metabolites showed correlation with the seizure frequency in the ipsilateral HR: ATPT, PME, PDE, PME/PDE. The seizure frequency was correlated to ?-ATP, ATPT, PCr/ ?-ATP, PCr/ATPT in the contralateral HR. Aditional analysis was made with the results from HR, dividing the patients according to seizure frequency and time of epilepsy. Patients with 10 or more seizures per month showed reduction of PME in the ipsilateral HR; increase of ?-ATP and decrease of PCr/?- ATP in the contralateral HR. Patients with time of epilepsy of 20 years or more showed increase of ?-ATP in the contralateral HR. Patients with less than 10 seizures per month or time of epilepsy lower than 20 years did not demonstrate any difference. Reduction of PME in the ipsilateral HR and tendency to significant differences in ?-ATP and PCr/?-ATP in the contralateral HR were detected from interpolated data. CONCLUSIONS: There are diffuse energetic changes in non epileptogenic regions of patients with MTS that could be related to mitochondrial dysfunction. In the ipsilateral HR, reduction of PME was found from interpolated data and in the group of patients with 10 or more seizures per month, suggesting changes in membrane composition, probably related to gliosis and neuronal loss