Philosophiae Doctor - PhD / Neurodegenerative disorders are characterised by progressive loss of the brain’s physiological functions as a result of gradual degeneration of neurons in the central nervous system. Even though they are classified as diseases of the elderly, occurrence earlier in life is possible, but that would suggest the influence of genetic and/or environmental factors. Due to the continuous rise in modernisation and industrialisation over the years, there has been an increase in incidence and prevalence of neurodegenerative disorders. With the advances in technology and life expectancy, the rates of the common forms (Alzheimer’s disease and Parkinson’s disease), are expected to increase exponentially by 2050. Unfortunately, there is still no clinically approved treatment or therapy to slow down or halt the degenerative process as most registered drugs only offer symptomatic relief. Confounding this issue is the lack of definite mechanism of neurodegeneration, which is still poorly defined and not completely understood. Nonetheless, the pathology of most neurodegenerative disorders is believed to be a combination of interrelated processes that eventually leads to neuronal cell death. Among the postulated processes, the impact of excitotoxicity mediated by NMDA receptor over-activation is prominent and it is implicated in virtually all neurodegenerative disorders. With this basic insight, it is believed that molecules capable of inhibiting NMDA receptors and associated calcium channels, without affecting the normal physiological functions of the brain, could potentially serve as good neuroprotective drugs. Competitive and uncompetitive blockers (MK-801 and ketamine) have been explored, but none were clinically accepted due to undesirable side effects such as hallucinations, sedation and depression. However, NGP1-01, a polycyclic cage molecule, has been shown to be neuroprotective through modulation of NMDA receptors and voltage gated calcium channels and attenuation of MPP+ -induced toxicity. A similar approach could be useful in the design and development of new neuroprotective drugs. The aim of this study was to synthesise a series of open and rearranged cage-like molecules and explore their neuroprotective potential in neuroblastoma SH-SY5Y cells. The proposed structures, with norbornane scaffolds that contained different moieties, were designed to structurally resemble NGP1-01 and MK-801. Once synthesised, the compounds were purified and characterised, and were evaluated for their biological activities. Compounds were first screened for cytotoxicity at different concentrations. Thereafter, they were evaluated for neuroprotective effects against MPP+ -induced excitotoxicity and for calcium flux modulatory effects on NMDA receptor and voltage gated calcium channels. The norbornane derivatives were synthesised and characterised, and all final products were afforded in sufficient yields. All compounds with the exception of two compounds displayed good cytotoxic profiles towards the SH-SY5Y neuroblastoma cells at 10 µM, 50 µM and 100 µM concentrations as they demonstrated percentage cell viabilities close to 100% (control treated cells). Only two compounds showed percentage cell viability of 51% and 59% at 100 µM. Utilising the same cell line, all compounds, tested at 10 µM, attenuated MPP+ -induced toxicity after 24 hours of exposure to a neurotoxin. This was evident in the 23% to 53% enhancement (significant with p < 0.05) in cell viability when compared to the MPP+ only treated cells. In comparison to known NMDA receptor and/or voltage gated calcium channel blockers (MK-801, NGP1-01 or nimodipine), the synthesised compounds demonstrated mono or dual inhibition of calcium channels as they effectively attenuated calcium influx by blocking NMDA receptors and/or voltage gated calcium channels expressed in neuroblastoma SHSY5Y cells. This group of compounds were found to be more potent NMDA receptor inhibitors, probably due to similarities with MK-801 and memantine, than voltage gated calcium channel inhibitors. All compounds demonstrated moderate to good calcium inhibitory effects at NMDA receptors in the range of 23% to 70% while a selected few displayed very little or no activity at the voltage gated calcium channels. In conclusion, 27 compounds with norbornane scaffolds were successfully synthesised and evaluated for cytotoxicity and neuroprotection. The abilities of the synthesised compounds to protect neurons from the neurotoxin MPP+ and reduce calcium flux into neuronal cells were successfully demonstrated. These characteristics are essential in neuroprotection as they may prove significant in halting or slowing down the disease progression. The compounds showing a good cytotoxicity profile, neuroprotective effects and ability to reduce calcium overload, could potentially act as neuroprotective agents with good safety profiles or contribute as lead structures to the development and design of structurally related molecules that could clinically benefit people with neurodegenerative disorders.

When the brain is exposed to a traumatic injury, the brain produces high amounts of neurosteroids like allopregnanolone and progesterone which show protective and neurogenic capacities. Alzheimer’s disease patients also have lower amounts of these neurosteroids in brain tissue. Neurosteroids act on GABAA receptors and cholesterol receptors which is interesting since both the cholesterol transporter ApoE and excitotoxicity seems to be issues plaguing the patients. To study if there is a relationship between Alzheimer’s disease and neurosteroids, there are ongoing phase one studies but neurobiological studies are equally important in order to understand the mechanism. In this work protective capabilities of allopregnanolone on induced toxicity was investigated in human neuroblastoma SH-SY5Y cells. Protection and induced toxicity were assessed by studying cell viability with MTT assay. Toxins used were the oxidative stress inducing agent t-BHP, excitotoxic glutamate and amyloid β25-35. Previous studies have found allopregnanolone to induce neurogenesis, decrease ROS levels, inhibit apoptosis and to have immunoregulatory capabilities. The present study did see an increase in cell viability when treated to 1x10-8 M allopregnanolone but this effect was not observed when the concentration was increased further to 1x10-7 M and 1x10-6 M. When the SH-SY5Y cells were treated with toxins after pretreatment of allopregnanolone, additional decrease was seen when compared to cells only treated with toxins. The present study discovered the influence of components like cell density and cell generation which is of value for researchers planning future neurobiological studies. These neurobiological studies give insight of the correct mechanisms in the brain, opening up opportunities for new efficient drugs to be developed.

This thesis consists of two parts: a report on the use of a binocular Shack-Hartmann (SH) sensor to study the dynamic correlation of ocular aberrations; and the application of an adaptive optics (AO) system to investigate the effect of the manipulation of aberrations on the accommodation control. The binocular SH sensor consists of one laser source and one camera to reduce system cost and complexity. Six participants took part in this study. Coherence function analysis showed that coherence values were dependent on the subject, aberration and frequency component. Inter-ocular correlations of the aberration dynamics were fairly weak for all participants. Binocular and monocular viewing conditions produced similar wavefront error dynamics. The AO system has a dual wavefront sensing channel. The extra sensing channel permits direct measurement of the eye¿s aberrations independent of the deformable mirror. Dynamic correction of aberrations during steady-state fixation did not affect the accommodation microfluctuations, possibly due to the prior correction of the static aberration level and/or the limited correction bandwidth. The inversion of certain aberrations during dynamic accommodation affected the gain and latency of accommodation response (AR), suggesting that the eye used the aberrations to guide its initial path of accommodative step response. Corrections of aberrations at various temporal locations of AR cycle produced subject- and aberration-dependent results. The gain and phase lag of the AR to a sinusoidally moving target were unaffected by aberration correction. The predictable nature of the target had been suggested as the reason for its failure to produce any significant effect on the AR gain and phase lag.

Les troubles neurodégénératifs, tels que la sclérose latérale amyotrophique (SLA) et la démence frontotemporale (DFT), ont été associés aux protéines de liaison à l'ARN (RBP). Les principales caractéristiques de la SLA sont l'agrégation d'une protéine de liaison à l'ARN appelée protéine de liaison TAR (TDP-43). Il a été démontré que TDP-43 se lie à G3BP1, un facteur de nucléation pour l'assemblage des granules de stress, pour le stabiliser. Les granules de stress sont des structures séparées par phases qui se forment dans des conditions stressantes et favorisent la survie cellulaire. Une altération de l’assemblage des granules de stress et une réduction du G3BP1 sont signalées dans la SLA. Cette réduction est due à un défaut dans les transcriptions codantes pour G3BP1 stabilisant TDP-43. Par conséquent, une réponse défaillante des granules de stress pourrait jouer un rôle majeur dans la maladie. Ainsi, ce projet de recherche se concentre sur la restauration de G3BP1, dont la déplétion est liée à la perte de fonction de TDP-43. En utilisant des composés de petites molécules identifiés lors d'une campagne de dépistage de médicaments, nous cherchons à augmenter l'expression de G3BP1, rétablissant ainsi le mécanisme SG endogène et favorisant la survie neuronale. La découverte de candidats principaux (NPX-047, NPX-000-115 et NPX-001-280) qui sauvent efficacement l'expression et la fonction de G3BP1 est prometteuse pour des thérapies potentielles contre la SLA. Ces composés ont été testés sur des cellules SHSY5Y traitées avec du si-TDP, mais aucune récupération de l'ARNm de G3BP1 n'a été observée malgré des niveaux plus élevés de signaux de luciférase. Ainsi, une enquête approfondie sur les divergences dans nos résultats constitue notre prochaine étape, ce qui n’a pas été possible pendant la durée limitée de cette mémoire. De plus, les cibles non ciblées de ces composés seront étudiées à l’aide du séquençage Bru Chase. Dans l’ensemble, cette étude explore de nouvelles stratégies pour restaurer l’expression de G3BP1, offrant ainsi une voie potentielle d’intervention thérapeutique dans la SLA. / Neurodegenerative disorders, such as Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Dementia (FTD), have been associated with RNA-binding proteins (RBPs). Major hallmark of ALS is aggregation of an RNA-binding protein called TAR binding protein (TDP-43). TDP-43 has shown to bind to G3BP1, a nucleating factor for stress granule assembly, to stabilize it. Stress granules (SGs) are phase separated structures that form under stressful conditions and promote cell survival. Impaired stress granules assembly and reduced G3BP1 is reported in ALS. This reduction is due to a defect in TDP-43 stabilizing G3BP1 encoding transcripts; thus, a failed stress granule response could have a major role in the disease. Thus, this research focuses on restoring G3BP1, whose depletion is linked to TDP-43 loss of function. By utilizing small-molecule compounds identified through a drug screening campaign, we seek to increase G3BP1 expression, consequently reinstating the endogenous SG mechanism and promoting neuronal survival. The discovery of lead candidates (NPX-047, NPX-000-115, and NPX-001-280) that effectively rescue G3BP1 expression and function offers promise for potential ALS therapies. These compounds were tested on SH-SY5Y cells treated with si-TDP however no rescue of G3BP1 mRNA was observed despite higher levels of luciferase signals. Thus, in-depth investigation of discrepancies in our results is our next step which was not possible during the limited timeline of this thesis. In addition, off-targets of these compounds will be investigated using BruChase-sequencing. Overall, this study explores novel strategies to restore G3BP1 expression, providing a potential avenue for therapeutic intervention in ALS.

No description available.

A Doença de Alzheimer (DA) é a principal causa de demência na população idosa e tende a se tornar um grave problema de saúde pública com o aumento da expectativa de vida da população mundial. A perda progressiva de memória, principal sintoma da demência em pacientes com DA, é atribuída a danos sinápticos e à perda neuronal desencadeadas pelo desequilíbrio entre a produção e a depuração do peptídeo A?. Evidências surgidas nos últimos 20 anos apontam os oligômeros solúveis de A? (A?O), produtos de agregação do peptídeo A?, como as principais espécies neurotóxicas na DA. Por conta disso, e também pela ausência de métodos diagnósticos pre-mortem e tratamento eficientes para essa demência, a busca por anticorpos conformacionais específicos para A?O está em ascensão. Testes clínicos com IgG anti-A? resultaram em efeitos colaterais inflamatórios mediados pela porção não variável Fc. Então, anticorpos conformacionais artificiais do tipo scFv, desprovidos de porção Fc, foram selecionados contra A?O. Dentre eles, está NUsc1, que é neuroprotetor contra A?O em cultura primária de neurônios. Neste trabalho, avaliamos a toxicidade de A?Os na linhagem de neuroblastoma humano SH-SY5Y diferenciada em neurônios maduros e comparamos a neuroproteção conferida por diferentes anticorpos contra A?Os, por ensaio de viabilidade celular com MTT. Também avaliamos a especificidade de NUsc1 por A?O comparativamente a lisozima monomérica e oligomérica em ensaio de ELISA, já que outros anticorpos conformacionais reconhecem epítopo compartilhado por estados oligoméricos de outras proteínas amiloidogênicas. Para a validação de células SH-SY5Y como modelo in vitro de neurônios maduros, a diferenciação foi induzida com ácido retinoico e BDNF e as células foram marcadas para as proteínas MAP2 e NeuN em ensaio de imunofluorescência. Células submetidas ao protocolo de diferenciação apresentaram aumento dos níveis dessas proteínas, mudança morfológica condizente com o esperado na maturação neuronal. Posteriormente, o desafio da cultura com A?O indicou morte celular dose-dependente e reversão desta morte segundo a dose administrada dos anticorpos 6E10 e NU-4. Obtivemos um sinal cerca de 400 vezes maior no reconhecimento de A?O por NUsc1 que para oligômeros de lisozima, quando presentes na mesma concentração, indicando forte especificidade de NUsc1 por A?O. Além disso, NUsc1 purificado em sistema de gelfiltração em HPLC não apresenta citotoxicidade em concentração equivalente a dos anticorpos 6E10 e NU-4 em ensaios de neuroproteção em cultura de SH-SY5Y diferenciada, sugerindo que, se NUsc1 for tão eficiente quanto estas IgG\'s, este poderá ser usado em dose não citotóxica. Portanto, podemos concluir que NUsc1 apresenta grande potencial como ferramenta diagnóstica e terapêutica para a DA, mas que mais experimentos para expandir sua validação e potencial ainda são necessários. / Alzheimer\'s Disease (AD) is the leading cause of dementia in the elderly population and tends to become a serious public health problem with increasing life expectancy of the world\'s population. Progressive memory loss, the main symptom of dementia in patients with AD, is attributed to synaptic damage and neuronal loss triggered by imbalance between production and clearance of the A? peptide. Evidence from the last 20 years indicates that soluble A? oligomers (A?O), A? peptide aggregation products, as the main neurotoxic species in AD. Because of this, and also because of the absence of efficient pre-mortem diagnostic and treatment methods for this dementia, the search for conformational antibodies specific for A?O is on the rise. Clinical tests with anti-A? IgG\'s resulted in inflammatory side effects mediated by the non-variable Fc portion. Then, artificial conformational antibodies of the scFv type, lacking the Fc portion, were selected against A?O. Among them is NUsc1, which is neuroprotective against A?O in primary neuronal culture. In this work, we evaluated the toxicity of A?Os in the differentiated SH-SY5Y human neuroblastoma line in mature neurons and compared the neuroprotection conferred by different antibodies against A?O types by MTT cell viability assay. We also evaluated the specificity of NUsc1 for A?O compared to monomeric and oligomeric lysozyme in the ELISA assay, since other conformational antibodies recognize epitope shared by oligomeric states of other amyloidogenic proteins. For the validation of SH-SY5Y cells as an in vitro model of mature neurons, differentiation was induced with retinoic acid and BDNF and the cells were labeled for MAP2 and NeuN proteins in immunofluorescence assay. Cells submitted to the differentiation protocol presented increased levels of these proteins, a morphological change consistent with the expected neuronal maturation. Subsequently, the challenge of culture with A?O indicated dose-dependent cell death and reversion of this death according to the administered dose of 6E10 and NU-4 antibodies. We obtained a 400-fold higher signal in the recognition of A?O by NUsc1 than for lysozyme oligomers, when present at the same concentration, indicating strong specificity of A?O by NUsc1. In addition, NUsc1 purified on HPLC gel-filtration system does not exhibit cytotoxicity at concentration equivalent to 6E10 and NU-4 antibodies in neuroprotection assays in differentiated SH-SY5Y culture, suggesting that, if NUsc1 is as efficient as these IgG\'s, it may be used in a non-cytotoxic dose. Therefore, we can conclude that NUsc1 presents great potential as a diagnostic and therapeutic tool for AD, but that further experiments to expand its validation and potential are still necessary.

A Doença de Alzheimer (DA) é a principal causa de demência na população idosa e tende a se tornar um grave problema de saúde pública com o aumento da expectativa de vida da população mundial. A perda progressiva de memória, principal sintoma da demência em pacientes com DA, é atribuída a danos sinápticos e à perda neuronal desencadeadas pelo desequilíbrio entre a produção e a depuração do peptídeo A?. Evidências surgidas nos últimos 20 anos apontam os oligômeros solúveis de A? (A?O), produtos de agregação do peptídeo A?, como as principais espécies neurotóxicas na DA. Por conta disso, e também pela ausência de métodos diagnósticos pre-mortem e tratamento eficientes para essa demência, a busca por anticorpos conformacionais específicos para A?O está em ascensão. Testes clínicos com IgG anti-A? resultaram em efeitos colaterais inflamatórios mediados pela porção não variável Fc. Então, anticorpos conformacionais artificiais do tipo scFv, desprovidos de porção Fc, foram selecionados contra A?O. Dentre eles, está NUsc1, que é neuroprotetor contra A?O em cultura primária de neurônios. Neste trabalho, avaliamos a toxicidade de A?Os na linhagem de neuroblastoma humano SH-SY5Y diferenciada em neurônios maduros e comparamos a neuroproteção conferida por diferentes anticorpos contra A?Os, por ensaio de viabilidade celular com MTT. Também avaliamos a especificidade de NUsc1 por A?O comparativamente a lisozima monomérica e oligomérica em ensaio de ELISA, já que outros anticorpos conformacionais reconhecem epítopo compartilhado por estados oligoméricos de outras proteínas amiloidogênicas. Para a validação de células SH-SY5Y como modelo in vitro de neurônios maduros, a diferenciação foi induzida com ácido retinoico e BDNF e as células foram marcadas para as proteínas MAP2 e NeuN em ensaio de imunofluorescência. Células submetidas ao protocolo de diferenciação apresentaram aumento dos níveis dessas proteínas, mudança morfológica condizente com o esperado na maturação neuronal. Posteriormente, o desafio da cultura com A?O indicou morte celular dose-dependente e reversão desta morte segundo a dose administrada dos anticorpos 6E10 e NU-4. Obtivemos um sinal cerca de 400 vezes maior no reconhecimento de A?O por NUsc1 que para oligômeros de lisozima, quando presentes na mesma concentração, indicando forte especificidade de NUsc1 por A?O. Além disso, NUsc1 purificado em sistema de gelfiltração em HPLC não apresenta citotoxicidade em concentração equivalente a dos anticorpos 6E10 e NU-4 em ensaios de neuroproteção em cultura de SH-SY5Y diferenciada, sugerindo que, se NUsc1 for tão eficiente quanto estas IgG\'s, este poderá ser usado em dose não citotóxica. Portanto, podemos concluir que NUsc1 apresenta grande potencial como ferramenta diagnóstica e terapêutica para a DA, mas que mais experimentos para expandir sua validação e potencial ainda são necessários. / Alzheimer\'s Disease (AD) is the leading cause of dementia in the elderly population and tends to become a serious public health problem with increasing life expectancy of the world\'s population. Progressive memory loss, the main symptom of dementia in patients with AD, is attributed to synaptic damage and neuronal loss triggered by imbalance between production and clearance of the A? peptide. Evidence from the last 20 years indicates that soluble A? oligomers (A?O), A? peptide aggregation products, as the main neurotoxic species in AD. Because of this, and also because of the absence of efficient pre-mortem diagnostic and treatment methods for this dementia, the search for conformational antibodies specific for A?O is on the rise. Clinical tests with anti-A? IgG\'s resulted in inflammatory side effects mediated by the non-variable Fc portion. Then, artificial conformational antibodies of the scFv type, lacking the Fc portion, were selected against A?O. Among them is NUsc1, which is neuroprotective against A?O in primary neuronal culture. In this work, we evaluated the toxicity of A?Os in the differentiated SH-SY5Y human neuroblastoma line in mature neurons and compared the neuroprotection conferred by different antibodies against A?O types by MTT cell viability assay. We also evaluated the specificity of NUsc1 for A?O compared to monomeric and oligomeric lysozyme in the ELISA assay, since other conformational antibodies recognize epitope shared by oligomeric states of other amyloidogenic proteins. For the validation of SH-SY5Y cells as an in vitro model of mature neurons, differentiation was induced with retinoic acid and BDNF and the cells were labeled for MAP2 and NeuN proteins in immunofluorescence assay. Cells submitted to the differentiation protocol presented increased levels of these proteins, a morphological change consistent with the expected neuronal maturation. Subsequently, the challenge of culture with A?O indicated dose-dependent cell death and reversion of this death according to the administered dose of 6E10 and NU-4 antibodies. We obtained a 400-fold higher signal in the recognition of A?O by NUsc1 than for lysozyme oligomers, when present at the same concentration, indicating strong specificity of A?O by NUsc1. In addition, NUsc1 purified on HPLC gel-filtration system does not exhibit cytotoxicity at concentration equivalent to 6E10 and NU-4 antibodies in neuroprotection assays in differentiated SH-SY5Y culture, suggesting that, if NUsc1 is as efficient as these IgG\'s, it may be used in a non-cytotoxic dose. Therefore, we can conclude that NUsc1 presents great potential as a diagnostic and therapeutic tool for AD, but that further experiments to expand its validation and potential are still necessary.

O vírus sincicial respiratório humano (VSRH) é o agente viral mais freqüentemente relacionado a doenças do trato respiratório inferior em crianças abaixo de um ano de idade. Analíse da varibilidade antigênica e gênica mostraram que o VSRH pode ser divido em dois grupos: A e B. O vírus é um membro do gênero Pneumovirus pertencente a família Paramyxoviridea, e possui três principais proteínas que são: glicoproteina F (fusão), glicoproteina G (adesão), glicoproteina SH (pequena proteína hidrofóbica). A proteína F é responsável pela fusão da célula ao vírus, enquanto a proteína G tem papel fundamental na replicação do vírus, porém a função da proteína SH, ainda não está bem definida, estudos recentes mostram-na como responsável por inibir a sinalização do fator de necrose tumoral alfa (TNF-a). Neste estudo foram colhidas amostras de 965 crianças, entre os anos de 2004 e 2005, dentre as quais 424 foram positivas. 117 amostras foram seqüenciadas a proteína SH e G e comparadas com amostras que circularam mundialmente. A analíse filogenética mostrou uma baixa variabilidade entre os genótipos estudados tanto do grupo A quanto do B. / The human respiratory syncytial virus (HRSV) is the major cause of lawer respiratory tract infections in infantis, young children and elderly. Analysis of the antigenic and genetic variability has shown that there are two groups of the virus HRSV, A and B. The virus (HRSV) is a member of the genus pneumovirus in the paramyxoviridae family. The virus encodes three membrane-bound glicoproteins, namely the fusion (F) attachment (G) and small hydrophobic (SH) proteins. The F mediates fusion of the virus and cell membranes and the G proteins is involved in virus attachment. The biological properties of the F and G glicoproteins and role that they play during virus replication relatively well understood, however the functional significance of the SH protein during replication remains unclear, although recent study shown that it can inhibit TNF-alpha. In this study, HRSV strains were isolated from nasopharyngeal aspirates collected from 965 children between 2004 and 2005, yielding 424 positive samples. We sequenced the small hydrophobic protein (SH) gene and protein (G) of 117 samples and compared them with other viruses identified worldwide. The phylogenetic analysis showed a low genetic variably among the isolates but allowed us to classify the viruses into different genotypes for the A and B HRSV strains.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Selenium (Se) is an oligoelement crucial for various biological processes. Se has anti-inflammatory and antioxidant properties and plays a key role in brain development. Adenosine deaminase (ADA, EC 3.5.4.4) is a key enzyme in purine metabolism because it helps in the regulation of intracellular and extracellular levels of adenosine, an important nucleoside that acts in the neuromodulation of the immune and nervous systems. Oxidative stress occurs when there is an imbalance between the production of reactive species and their detoxification by systems that remove or repair the resulting damage. In this context, several Se compounds have been developed and studied, among them the 3-methyl-1-phenyl-2-(phenylseleno)oct-2-en-1-one (C12H2HOSe), which is an α, β-unsaturated ketone functionalized vinyl chalcogenide, and sodium selenate (Na2SeO4), the main inorganic form of Se found in animals and plants. The aim of this study was to compare the effects of C12H2HOSe (organic Se) and Na2SeO4 (inorganic Se) on the cell viability, lipoperoxidation and ADA activity in cortex slices of young rats. The results showed that only organic Se caused a reduction in ADA activity at the concentrations of 1, 10 and 30μM in the cerebral cortex slices of the rats tested. However, this result is not related to the oxidation of the thiol groups, since this compound did not alter the NP-SH and LDH levels. Both compounds did not affect the lipoperoxidation levels, although the organic Se was capable of sequestering NO radicals at the highest concentration tested. The inorganic Se protected the cerebral cortex slices against the sodium nitroprusside-induced damage and increased the NP-SH levels. The tests used to evaluate cell viability (LDH and MTT) suggest the maintenance of the cell integrity of the cortex slices exposed to Se compounds. Therefore, our results suggest that organic Se has immunomodulatory properties, due to the reduction in ADA activity, and acted in the maintenance of the cell integrity. In turn, the antioxidant activity of inorganic Se was reaffirmed. Hence, the results of this study paved the way to explore both Se compounds in the CNS. / O selênio (Se) é um oligoelemento essencial em vários processos biológicos. Possui propriedades antiinflamatórias, antioxidantes e desempenha um papel fundamental no desenvolvimento do cérebro. A adenosina deaminase (ADA, E.C. 3.5.4.4) é uma enzima chave no metabolismo das purinas, pois auxilia na regulação dos níveis intra e extracelulares de adenosina, um importante nucleosídeo que atua na neuromodulação dos sistemas nervoso e imune. O estresse oxidativo ocorre quando há desequilíbrio entre a produção de espécies reativas e sua desintoxicação através de sistemas que removem ou reparem os danos por elas causados. Nesse contexto, vários compostos de Se vêm sendo desenvolvidos e estudados, entre eles podemos citar o 3-metil-1-fenil-2-(fenilseleno)oct-2-en-1-um(C12H2HOSe),um organocalcogênio com uma cetona α, β- insaturada funcionando como um vinil calcogênio e o selenato de sódio (Na2SeO4), principal Se inorgânico encontrado em animais e plantas. Este estudo teve como objetivo comparar os efeitos do C12H2HOSe (Se orgânico) e Na2SeO4 (Se inorgânico) na viabilidade celular, parâmetros relacionados ao estresse oxidativo e atividade da ADA em fatias de córtex de ratos jovens. Os resultados demonstraram que apenas o Se orgânico provocou a diminuição na atividade da ADA nas concentrações de 1, 10 e 30μM nas fatias de córtex cerebral dos ratos testados. No entanto, esse resultado não tem relação com a oxidação de seus grupamentos tióis, já que esse composto não alterou os níveis de NP-SH e de LDH. Apesar de ambos os compostos não alteraram os níveis de lipoperoxidação, o Se orgânico apresentou capacidade de sequestrar radicais NO na maior concentração testada. Já o composto inorgânico de Se protegeu as fatias de córtex cerebral contra o dano induzido pelo nitroprussiato de sódio e aumentou os níveis de NP-SH. Os testes de viabilidade celular (LDH e MTT) sugerem a manutenção da integridade celular em fatias de córtex de ratos jovens quando expostos aos compostos de Se. Assim, nossos resultados sugerem que o Se orgânico apresenta propriedades imunomoduladoras por reduzir a atividade da ADA, além de atuar na manutenção da integridade celular. Já o Se inorgânico teve sua atividade antioxidante confirmada. Portanto, os resultados obtidos neste estudo destacam um caminho promissor a ser explorado por ambos os compostos no SNC.

Des marqueurs biologiques exogènes à base de nanoparticules hybrides possédant des propriétés optiques non-linéaires ont été préparés en tant qu'agent de contraste pour l'imagerie de second harmonique et le diagnostic de cellules pathogènes. L'iodate de fer est l'un des matériaux proposés dans ce domaine en raison de ses bonnes propriétés de second harmonique et de sa faible toxicité. Des nanoparticules d'iodate de fer ont été synthétisées par microémulsion inverse et miniémulsion inverse. Un bon contrôle de taille et de morphologie des particules a été obtenu dans les deux cas. En vue d'applications biomédicales, l'encapsulation des particules a été réalisée in situ par polymérisation en miniémulsion inverse. Enfin, l'encapsulation de particules de niobate de potassium par un polymère biocompatible a été étudiée. L'intérêt de ces particules hybrides a été démontré par des études in vitro en imagerie de second harmonique. / Exogenous biomarkers based on hybrid nanoparticles with nonlinear optical properties were prepared as a contrast agent for second harmonic imaging and diagnosis of pathogenic cells. Iron iodate is one of the selected materials for this specific field due to its good second harmonic properties and its low toxicity. Iron iodate nanoparticles were synthesized by inverse microemulsion and inverse miniemulsion. In both cases, a good control of size and morphology was achieved. For biomedical applications, nanoparticles encapsulation was carried out in situ by reverse miniemulsion polymerization. Finally, encapsulation of potassium niobate nanoparticles with a biocompatible polymer was conducted. Their interest was demonstrated in vitro by second harmonic imaging studies.