Submitted by admin tede (tede@pucgoias.edu.br) on 2018-05-03T12:58:27Z No. of bitstreams: 1 Husys Cardoso Souto.pdf: 1103920 bytes, checksum: 1a37590505c8ec6568fe48b8acbb758f (MD5) / Made available in DSpace on 2018-05-03T12:58:27Z (GMT). No. of bitstreams: 1 Husys Cardoso Souto.pdf: 1103920 bytes, checksum: 1a37590505c8ec6568fe48b8acbb758f (MD5) Previous issue date: 2018-03-15 / Introduction: Physiotherapeutic rehabilitation and the Takkyu Volley therapeutic workshop are two very important modalities for the treatment of neurological patients. Objective: To evaluate the impact of conventional physiotherapy treatment and the Takkyu Volley therapeutic workshop on the functional independence and quality of life of patients in neurological rehabilitation. Method: This is a longitudinal, prospective, analytical study with 30 patients, performed at the Rehabilitation Clinic, from 2016/June to 2017/November, in Goiânia/GO. Of these patients, 10 were submitted only to conventional physiotherapy and 20, besides conventional physiotherapy, were also directed to the Takkyu Volley therapeutic workshop. Three questionnaires were applied: sociodemographic, SF-36v2 quality of life and Barthel Index. Descriptive and comparative statistics were performed. Results: The evaluated group had a mean age of 52.4 years; it consisted mainly of men, lived with the spouse, with family income up to a minimum wage. The evaluation of the quality of life showed greater impact on the dimensions "Functional Capacity", "General Health Status" and "Pain". The Takkyu Volley Therapy Workshop excelled at some items compared to conventional physiotherapy. In the first phase they achieved better scores in the dimensions: mental component, social aspect. And in the second phase: physical component, mental component, functional capacity, physical aspect, general state of health and emotional aspect. All the data evaluated by the Barthel index questionnaire did not show statistically significant differences in the comparison between the two modalities evaluated. Conclusion: Both treatments had positive results in improving functional disability and patients' quality of life, with special emphasis on the Takkyu Volley workshop, which achieved more expressive results. / Introdução: A reabilitação fisioterapêutica e a oficina terapêutica Takkyu Volley são duas modalidades importantíssima para o tratamento de pacientes neurológicos. Objetivo: Avaliar o impacto do tratamento de fisioterapia convencional e da oficina terapêutica Takkyu Volley na independência funcional e na qualidade de vida dos pacientes em reabilitação neurológica. Método: Trata-se de um estudo analítico, longitudinal, prospectivo, com 30 pacientes, realizado na Clínica Reabilitar, no período de junho/2016 a novembro/2017, em Goiânia/GO. Deste pacientes, 10 foram submetidos apenas à fisioterapia convencional e 20, além da fisioterapia convencional, foram também direcionados à oficina terapêutica Takkyu Volley. Foram aplicados três questionários: sociodemográfico, de qualidade de vida SF-36v2 e Índice de Barthel. Foram realizadas estatísticas: descritiva e comparativa. Resultados: O grupo avaliado tinha a média de idades de 52,4 anos, era constituído principalmente de homens, morava com o cônjuge, com renda familiar de até um salário mínimo. A avaliação da qualidade de vida apontou maior impacto sobre as dimensões "Capacidade Funcional", "Estado Geral de Saúde" e "Dor". A Oficina Terapêutica Takkyu Volley sobressaiu em alguns itens em relação à fisioterapia convencional. Na primeira fase alcançaram melhores escores nas dimensões: componente mental, aspecto social. E na segunda fase: componente físico, componente mental, capacidade funcional, aspecto físico, estado geral de saúde e aspecto emocional. Todos os dados avaliados pelo questionário do índice de Barthel não mostraram diferenças estatisticamente significantes na comparação entre as duas modalidades avaliadas. Conclusão: Ambos os tratamentos tiveram resultados positivos na melhorar a incapacidade funcional e da qualidade de vida dos pacientes, com destaque especial para a oficina Takkyu Volley, que atingiram resultados mais expressivos.

When adults with neurological conditions can access occupational therapy (OT) services, they have better outcomes. Unfortunately, many people who are living in the community with these conditions do not participate in OT services. This can compound deficits, lead to re-hospitalization, and negatively impact independence and quality of life. The issues with access for this population, specifically adults aged 18-65, can largely be attributed to lack of awareness of the role and scope of OT, lack of availability of specialized services, and issues with affordability. Evolve Occupational Therapy (Evolve OT) is a program designed specifically for this group, adults with neurological conditions aged 18-65 who have difficulty accessing OT services. This innovative approach to service delivery provides an alternative access pathway, treatment, and payment models that are intentionally designed to increase participation in and utilization of OT services. With this program, it is expected that patients will report increased satisfaction with participation, increased quality of life, and outcomes will hopefully effect change amongst policymakers.

Muscle weakness is one of the major causes of post-stroke disability. Stroke rehabilitation programs now often incorporate the same type of resistance training that is used for healthy subjects; however, the training effects induced from these training strategies are often limited for stroke patients. An important resistance training principle is that an optimal level of stress is exerted on the neuromuscular system, both during concentric (shortening) and eccentric (lengthening) contractions. One potential problem for post-stroke patients might be difficulties achieving sufficient levels of stress on the neuromuscular system. This problem may be associated with altered muscular function after stroke. In healthy subjects, maximum strength during eccentric contractions is higher than during concentric contractions. In individuals with stroke, this difference in strength is often increased. Moreover, it has also been shown that individuals with stroke exhibit alteration with respect to how the strength varies throughout the range of motion. For example, healthy subjects exhibit a joint specific torque-angle relationship that normally is the same irrespective of contraction mode and contraction velocity. In contrast, individuals with stroke exhibit an overall change of the torque-angle relationship. This change, as described in the literature, consists of a more pronounced strength loss at short muscle length. In individuals with stroke, torque-angle relationships are only partially investigated and so far these relationships have not been analysed using testing protocols that include eccentric, isometric, and concentric modes of contraction.   This thesis investigates the torque-angle relationship of elbow flexors in subjects with stroke during all three modes of contractions – isometric, concentric, and eccentric ­– and the relative loading throughout the range of movement during a resistance exercise. In addition, this thesis studies possible central nervous system mechanisms involved in the control of muscle activation during eccentric and concentric contractions.   The torque-angle relationship during maximum voluntary elbow flexion was examined in stroke subjects (n=11), age-matched healthy subjects (n=11), and young subjects (n=11) during different contraction modes and velocities. In stroke subjects, maximum torque as well as the torque angle relationship was better preserved during eccentric contractions compared to concentric contractions. Furthermore, the relative loading during a resistance exercise at an intensity of 10RM (repetition maximum) was examined. Relative loading throughout the concentric phase of the resistance exercise, expressed as percentage of concentric torque, was found to be similar in all groups. However, relative loading during the eccentric contraction phase, expressed as the percentage of eccentric isokinetic torque, was significantly lower for the stroke group. In addition, when related to isometric maximum voluntary contraction, the loading for the stroke group was significantly lower than for the control groups during both the concentric and eccentric contraction phases. Functional magnetic resonance imaging was used to examine differences between recruited brain regions during the concentric and the eccentric phase of imagined maximum resistance exercise of the elbow flexors (motor imagery) in young healthy subjects (n=18) and in a selected sample of individuals with stroke (n=4). The motor and premotor cortex was less activated during imagined maximum eccentric contractions compared to imagined maximum concentric contraction of elbow flexors. Moreover, BA44 in the ventrolateral prefrontal cortex, a brain area that has been shown to be involved in inhibitory control of motor activity, was additionally recruited during eccentric compared to concentric conditions. This pattern was evident only on the contralesional (the intact hemisphere) in some of the stroke subjects. On the ipsilesional hemisphere, the recruitment in ventrolateral prefrontal cortex was similar for both modes of contractions.    Compared to healthy subjects, the stroke subjects exhibited altered muscular function comprising a specific reduction of torque producing capacity and deviant torque-angle relationship during concentric contractions. Therefore, the relative training load during the resistance exercise at a training intensity of 10RM was lower for subjects with stroke. Furthermore, neuroimaging data indicates that the ventrolateral prefrontal cortex may be involved in a mechanism that modulates cortical motor drive differently depending on mode of the contractions. This might partly be responsible for why it is impossible to fully activate a muscle during eccentric contractions. Moreover, among individuals with stroke, a disturbance of this system could also lie behind the lack of contraction mode-specific modulation of muscle activation that has been found in this population. The altered neuromuscular function evident after a stroke means that stroke victims may find it difficult to supply a sufficient level of stress during traditional resistance exercises to promote adaptation by the neuromuscular system. This insufficiency may partially explain why the increase in strength, in response to conventional resistance training, often has been found to be low among subjects with stroke. / Muskelsvaghet är en av orsakerna till funktionshinder efter stroke. I rehabiliteringsprogram för personer som drabbats av stroke förekommer det numera att styrketräning används i syfte att öka muskelstyrkan. Effekten av styrketräning har dock ofta visat sig vara begränsad. En viktig styrketräningsprincip är att muskulaturen belastas tillräckligt nära maximal styrka under både koncentriska kontraktioner (när man lyfter en vikt) och excentriska kontraktioner (när man kontrollerat sänker en vikt). Ett potentiellt problem skulle kunna vara att personer med stroke inte belastas optimalt under träning på grund av förändrad muskelfunktion. Efter stroke är muskelfunktionen ofta förändrad såtillvida att styrkenedsättningen är mer uttalad under koncentriska kontraktioner. Därutöver har man funnit att styrkenedsättningen är mest uttalad när muskeln är i sitt mest förkortade läge. Detta fenomen har dock inte studerats för alla tre kontraktionstyper, det vill säga excentriska, koncentriska och isometriska kontraktioner, hos personer med stroke.   Denna avhandling undersöker sambandet mellan styrka och ledvinkel över armbågsleden hos personer med stroke under alla tre kontraktionstyper – excentrisk, koncentrisk och isometrisk, samt relativ belastning genom rörelsebanan under en styrketräningsövning. Därutöver undersöker denna avhandling också hjärnans aktiveringsmönster under excentriska och koncentriska kontraktioner.   Sambandet mellan styrka och ledvinkel undersöktes hos personer med stroke (n = 11), åldersmatchade (n = 11) och unga försökspersoner (n = 11). Jämfört med kontrollgrupperna var maximal styrka för personer med stroke mest nedsatt, samt även den oproportionerligt stora styrkenedsättningen vid kort muskelängd som mest uttalad, under koncentriska kontraktioner. Denna avvikelse var minst uttalad vid excentriska kontraktioner. Vidare studerades hur hög belastningen på muskulaturen var i jämförelse med muskelns maximala styrka under en styrketräningsliknande övning för armbågsflexorer vid en träningsintensitet på 10RM. Den uppmätta belastningen under den koncentriska fasen av styrketräningsövningen, uttryckt som procent av den genomsnittliga koncentriska styrkan, var densamma för alla grupperna. Under den excentriska fasen av övningen var dock belastningen, uttryckt som procent av den maximala excentriska styrkan, signifikant lägre för personer med stroke. Träningsbelastningen utgjorde också en lägre andel av den maximala isometriska styrkan för personer med stroke, både under den koncentriska och under den excentriska fasen.   Funktionell magnetresonanstomografi (fMRI) användes för att undersöka hjärnans aktiveringsmönster hos unga försökspersoner (n = 18) och hos individer med stroke (n = 4) när de föreställde sig att de utförde maximal styrketräning för armbågsflexorer (motor imagery). Resultatet visade att primära motorbarken och premotoriska barken var mindre aktiverade när unga friska försökspersonerna föreställde sig utföra maximala excentriska, jämfört med maximala koncentriska kontraktioner. Dessutom var en region i ventrolaterala prefrontala barken, som i tidigare studier visat sig vara inblandat i reglering och hämning av muskelaktivering, mer aktiverade under föreställda excentriska kontraktioner. Detta aktiveringsmönster i den prefrontala barken återfanns dock endast i den icke skadade hjärnhalvan hos personer med stroke.   Jämfört med kontrollgrupperna uppvisade försökspersonerna med stroke en förändrad muskelfunktion som bestod av en specifik nedsättning av styrkan under koncentriska kontraktioner samt också ett mer avvikande samband mellan styrka och ledvinkel under koncentriska kontraktioner. Den relativa belastningen under utförandet av en styrketräningsövning med en intensitet på 10RM var på grund av dessa avvikelser lägre för försökspersoner med stroke. Hjärnavbildnings-studierna indikerade att ventrolaterala prefrontala barken verkar vara involverat i ett kortikalt moduleringssystem som reglerar muskel-aktivering olika beroende på kontraktionstyp under maximala kontraktioner. Detta skulle kunna vara en underliggande mekanism bakom den hittills obesvarade frågan varför det är omöjligt att aktivera muskulaturen maximalt under excentriska kontraktioner. En störning av detta moduleringssystem hos personer med stroke verkar också kunna ligga bakom den förändrade regleringen av muskelaktivering som visat sig förekomma hos personer med stroke. Neuromuskulär funktion efter stroke är förändrad i flera avseenden vilket verkar medföra att muskulaturen inte belastas optimalt under konventionell styrketräning. Detta kan vara en delförklaring till varför styrkeökningen som svar på träning ofta är liten hos personer med stroke.

The Ytterby mine is located on Resarö island in the Stockholm archipelago. Mainly feldspars but also quartz were historically quarried in the mine, which is also the place of discovery of seven rare earth elements (REE). During the cold war era, the mine shaft was used as a diesel and jet fuel deposit for the Swedish Armed Forces. Recently, a spatial coupling between multiple sclerosis (MS), a chronic neurodegenerative disease in the central nervous system, prevalence and the quarry has been suggested. Previous studies show that adverse neurological health effects are associated with oral intake of REEs and there is support for a coupling between ionizing radiation and MS. The extent and character of health effects as a result of exposure to petroleum products are still debated. However, a substantial number of scientific reports support a coupling between neurodegenerative health effects and toxic constituents of jet fuels such as benzene, toluene, and n-hexane. My data show that a possible overrepresentation of MS patients within the Ytterby postal code area could be an indication of a spatial coupling between the mine and MS. Such a possible coupling could be associated with the REEs present in the local rocks, with the previous storage of diesel or jet fuel MC-77 in the mine and/or with zones of high natural radioactivity in the area. Water samples collected in 15 wells in the Ytterby village show traces of five REEs, i.e. scandium (Sc), yttrium (Y), lanthanum (La), neodymium (Nd) and samarium (Sm) and the majority of sample locations at low ground elevation show contamination of diesel which is the most recent fuel stored in the mine. Moreover, results from an analysis of a black substance leaking out of cracks in the mine corridors confirm that REEs are present in substantial concentrations in the local rocks and also appear to be mobile. This should be taken into account when considering a potential contamination of the local water supply. Measurements of natural radioactivity have also been made around the contours of the quarry and zones of high ionizing radiation have been identified. By using these zones of high ionizing radiation as a proxy for rare minerals containing rare earth elements, I further suggest that the REE occurrences are highly localized around the quarry and could be associated with, or remobilized by, younger faults.  My data show that a full investigation is warranted of a possible spatial coupling between neurological health issues, MS being one of them, and the mine.

Antibodies directed against AMPAR and GABA_BR subunits have been implicated in forms of limbic encephalitis (LE), a disease characterised by memory loss and seizures. Patients with LE show clinical improvement with immunomodulatory treatment, suggesting that the associated antibodies are pathogenic. To explore further the AMPAR and GABA_BR antibodies, an in house cell based assay (CBA) was established for screening and potential pathogenicity was explored using a series of in vitro experiments. Human embryonic kidney (HEK) cells transfected with AMPAR and GABA_BR subunits and primary neuronal cultures were used to detect antibodies in patient sera and CSF. In total, 15/1361 (1.1%) AMPAR antibody positive samples and 24/1438 (1.7%) GABABR antibody positive samples were identified. The predominant antibody subclass for AMPAR and GABA_BR antibodies was shown to be IgG1. Interestingly, on transfected cells, only AMPAR antibodies showed complement deposition, and therefore had the potential to activate the classical pathway of the complement cascade. Application of IgG purified from AMPAR antibody positive patients, but not GABA_BR antibody positive patients caused a down regulation of the receptor from the cell surface of transfected HEK cells and primary hippocampal cultures. Electrophysiological analysis showed changes in Up state duration and spike rate in the entorhinal cortex following application of purified GABA_BR antibody IgG on brain slices. These findings suggest that GABABR antibodies are having a direct short term effect on GABA_BRs, and by extension cortical networks. Finally, we attempted to study whether or not viral infection could be a trigger for antibody production to known and novel antigen targets in a cohort of Japanese encephalitis viral (JEV) samples. A JEV cohort of 44 children was screened for antibodies to neuronal surface proteins by CBA. Twenty-seven percent of patients had antibodies to known neuronal surface antigens, with NMDAR and CASPR2 as the most common antigen targets. Interestingly, screening the cohort on primary neuronal cultures revealed that 65.9% of children with JEV have neuronal surface antibodies. The identification of the novel antigen target was attempted using immunoprecipitation and mass spectrometry techniques. In total, 4 neuronal surface proteins were identified that could be potential targets in JEV patients. In summary, antibodies directed against previously described antigenic targets were explored further for pathogenic potential, and a cohort of patients with a viral infection with potential novel antigen targets was investigated.

In 2009, the Government of Canada announced a four year national population health study on neurological conditions. The aim of the study was divided into four focus areas: incidence and prevalence of neurological conditions (scope of problem); risk factors for developing neurological conditions; health services, including gaps in services; and the impacts of neurological conditions. The Native Women’s Association of Canada (NWAC), with Dr. Carrie Bourassa, First Nations University of Canada, as the principal investigator, submitted a proposal to look at three out of the four focus areas, risk factors, health services / health gaps, and impacts, among Indigenous women. Out of the 13 research projects that were funded, this was the only project that focused specifically on Indigenous people, gathering much needed baseline information on how Indigenous people think about neurological conditions, how it impacts their lives, their families, and communities, and what they see as needed to support neurological health and wellbeing. Individual interviews and research circles were conducted with people who live with a neurological condition and caregivers of people with a neurological condition. Key informant interviews were also conducted with traditional knowledge keepers, health care professionals and practitioners. The open ended questions encouraged participants to share as much or as little information as they wanted to. The stories shared contained a wealth of information, far exceeding the study’s focus areas. Unfortunately, due to external deadlines and budgetary constraints, the research team only had time to focus the research report on the three key areas outlined in the proposal–risk factors, health gaps, and impacts. A lot of the information shared was not fully explored. In this dissertation, a secondary analysis of the data is conducted to explore role of culture, as well as cultural understandings of neurological conditions, and interactions with the health care system. The theoretical framework will utilize Indigenous ways of knowing and Critical Medical Anthropology as part of a "two-eyed seeing" approach. Mi'kmaw Elder Albert Marshall suggested the phrase "two eyed seeing" as a guiding principle for health research, where one eye looks at the issue through the strengths of Indigenous knowledges and ways of knowing, while the other eye looks at the issue from the strengths of Western knowledges and ways of knowing. By using both eyes together to fully analyse the material, the strengths of both Indigenous and Western knowledges are brought together. Through using these different frameworks to explore the narratives, the research fills a gap in the literature regarding how Indigenous cultural understandings of neurological conditions can influence how Indigenous people access care.

<p>Human genetics has been experiencing a wave of genetic discoveries thanks to the development of several technologies, such as genome-wide association studies (GWAS), whole-exome sequencing, and whole genome sequencing. Despite the massive genetic discoveries of new variants associated with human diseases, several key challenges emerge following the genetic discovery. GWAS is known to be good at identifying the locus associated with the patient phenotype. However, the actually causal variants responsible for the phenotype are often elusive. Another challenge in human genetics is that even the causal mutations are already known, the underlying biological effect might remain largely ambiguous. Functional evaluation plays a key role to solve these key challenges in human genetics both to identify causal variants responsible for the phenotype, and to further develop the biological insights from the disease-causing mutations. </p><p>We adopted various methods to characterize the effects of variants identified in human genetic studies, including patient genetic and phenotypic data, RNA chemistry, molecular biology, virology, and multi-electrode array and primary neuronal culture systems. Chapter 1 is a broader introduction for the motivation and challenges for functional evaluation in human genetic studies, and the background of several genetics discoveries, such as hepatitis C treatment response, in which we performed functional characterization. </p><p>Chapter 2 focuses on the characterization of causal variants following the GWAS study for hepatitis C treatment response. We characterized a non-coding SNP (rs4803217) of IL28B (IFNL3) in high linkage disequilibrium (LD) with the discovery SNP identified in the GWAS. In this chapter, we used inter-disciplinary approaches to characterize rs4803217 on RNA structure, disease association, and protein translation.</p><p>Chapter 3 describes another avenue of functional characterization following GWAS focusing on the novel transcripts and proteins identified near the IL28B (IFNL3) locus. It has been recently speculated that this novel protein, which was named IFNL4, may affect the HCV treatment response and clearance. In this chapter, we used molecular biology, virology, and patient genetic and phenotypic data to further characterize and understand the biology of IFNL4. The efforts in chapter 2 and 3 provided new insights to the candidate causal variant(s) responsible for the GWAS for HCV treatment response, however, more evidence is still required to make claims for the exact causal roles of these variants for the GWAS association. </p><p>Chapter 4 aims to characterize a mutation already known to cause a disease (seizure) in a mouse model. We demonstrate the potential use of multi-electrode array (MEA) system for the functional characterization and drug testing on mutations found in neurological diseases, such as seizure. Functional characterization in neurological diseases is relatively challenging and available systematic tools are relatively limited. This chapter shows an exploratory research and example to establish a system for the broader use for functional characterization and translational opportunities for mutations found in neurological diseases. </p><p>Overall, this dissertation spans a range of challenges of functional evaluations in human genetics. It is expected that the functional characterization to understand human mutations will become more central in human genetics, because there are still many biological questions remaining to be answered after the explosion of human genetic discoveries. The recent advance in several technologies, including genome editing and pluripotent stem cells, is also expected to make new tools available for functional studies in human diseases.</p> / Dissertation

Apesar da introdução da HAART, formas leves de Transtornos Neurocognitivos Associados ao HIV (HAND) permanecem altamente prevalentes, afetando metade de todos os indivíduos infectados. A inflamação sistêmica e localizada induzida pelo HIV é considerada um dos mecanismos da HAND, e embora muitos biomarcadores potenciais tenham sido estudados, até o momento nenhum deles provou ser útil na prática clínica. Portanto, o objetivo desse trabalho foi investigar os níveis de biomarcadores de ativação celular, neurodegeneração e virológicos, no líquido cefalorraquidiano (CSF), e também marcadores genéticos no sangue, buscando associá-los à presença de HAND. Materiais e métodos: Utilizando um desenho transversal, níveis dos marcadores de ativação celular sCD14, Neopterina, MCP-1, IL-1b, IL-6, TNF-?, CXCL-10, IFN-? e MIP-1?; marcadores neuronais Tau, p-Tau, A?40, A?42 e Neurofilamentos; carga viral de HIV e níveis ApoE foram mensurados em 84 amostras de LCR , e a genotipagem do vírus, bem como a genotipagem da ApoE, foram feitas no sangue de 33 indivíduos infectados pelo HIV com alteração neurocognitiva assintomático (ANI), 15 com alteração neurocognitiva de leve a moderada (MND), 15 com demência associada ao HIV ( HAD), 14 controles infectados pelo HIV (C HIV +) e 7 controles não infectados pelo HIV (C HIV-). Os dados clínicos também foram avaliados. Resultados: O parâmetro de idade diferiu estatisticamente entre os grupos, por isso foi ajustado para análise posterior. Os controles HIV+ e o grupo HAND estavam todos sob HAART e aproximadamente 96% deles apresentaram carga viral plasmática e liquórica suprimidas. Os marcadores de neurodegeneração não diferiram estatisticamente entre os grupos. No entanto, os marcadores de ativação celular IFN-gama, IL-1beta e sCD14, juntamente com a ApoE, mostraram diferenças significativas. A análise discriminatória revelou que ApoE e IL-1? juntos possuem maior poder discriminatório para diferenciar o grupo HAND dos controles HIV+. A IL-1b mostrou um aumento progressivo no declínio cognitivo, enquanto os níveis de ApoE mostraram-se mais elevados nos controles HIV+, grupo que também teve a maior porcentagem de genótipo ?4. sCD14 por sua vez mostrou-se elevado no grupo HAND, enquanto IFN-? apresentou queda. Conclusão: Os resultados reforçam o conceito de que a elevação da regulação das citocinas pró-inflamatórias, como sCD14 e IL-1beta, pode desempenhar um papel na patogênese da HAND, mesmo nos pacientes com supressão viral.Em contrapartida, nenhum marcador de neurodegeneração apresentou diferença estatística entre os grupos. É possível que as diferenças encontradas em relação a ApoE nos grupos, tanto tem termos de regulação como a presença do genótipo e4, indique algum mecanismo compensatório, que poderia resultar em um fator protetor contra HAND, portanto deve ser melhor estudado. / Despite the introduction of HAART, mild forms of HIV-associated Neurocognitive Disorders (HAND) remain highly prevalent, affecting half of all infected individuals. Systemic and localized inflammation induced by HIV is considered to be one of the mechanisms of HAND, and although many potential biomarkers have been studied, none of them have proven to be useful in clinical practice. Therefore, the objective of this study was to investigate the levels of biomarkers of cellular activation, neurodegeneration and virological, in the cerebrospinal fluid (CSF), as well as genetic markers in the blood, seeking to associate them with the presence of HAND. Materials and methods: Using a cross-sectional design, levels of the cell activation markers sCD14, Neopterin, MCP-1, IL-1b, IL-6, TNF-?, CXCL-10, IFN-? and MIP-1?; neuronal markers Tau, p-Tau, A?40, A?42 and Neurofilaments; HIV viral load and ApoE levels were measured in 84 CSF samples, and virus genotyping and ApoE genotyping were done in the blood of 33 HIV-infected individuals with asymptomatic neurocognitive impairment (ANI), 15 with neurocognitive impairment (MND), 15 with HIV-associated dementia (HAD), 14 HIV-infected controls (C HIV +), and 7 non-HIV-infected controls (C HIV-). Clinical data were also evaluated. Results: The age parameter differed statistically between the groups, so it was adjusted for later analysis. HIV + controls and HAND group were all under HAART and approximately 96% of them had suppressed plasma and CSF viral load. Neurodegeneration markers did not differ statistically between the groups. However, the cell activation markers IFN-gamma, IL-1beta and sCD14, along with ApoE, showed significant differences. Discriminatory analysis revealed that ApoE and IL-1? together have greater discriminatory power to differentiate HAND group from HIV + controls. IL-1b showed a progressive increase in cognitive decline, while ApoE levels were higher in HIV + controls, which also had the highest percentage of ?4 genotype. sCD14 in turn showed to be elevated in the HAND group, while IFN-? showed decrease. Conclusion: The results reinforce the concept that increased regulation of proinflammatory cytokines, such as sCD14 and IL-1beta, may play a role in the pathogenesis of HAND, even in patients with viral suppression. On the other hand, no neurodegeneration marker presented statistical difference between groups. It is possible that the differences found regarding ApoE in the groups, both in terms of regulation and the presence of the e4 genotype, indicate some compensatory mechanism, which could result in a protective factor against HAND, so it should be better studied.

Reliable biomarkers of axonal damage are urgently needed in neurological diseases. Neurofilaments (Nf) are specific structural elements of neurons composed of at least three subunits: Nf light chain (NfL), Nf medium and Nf heavy chain (NfH). This PhD aimed to characterise NfL levels and their correlation with clinical features in patients with neurological diseases with a different rate of progression and following and under different treatment regimes. An important aim was also to develop a bioassay for NfL measurements in blood. Cerebrospinal fluid (CSF) NfL levels discriminated patients with a clinically isolated syndrome (CIS) (p=0.001) or multiple sclerosis (MS) (p=0.035) from healthy controls more efficiently, and was more sensitive to change after natalizumab therapy (p<0.0001) than CSF NfH (p=0.002). Further, CSF NfL levels decreased in fingolimodtreated MS patients (p=0.001), but not in those receiving placebo (p=0.433). Based on these findings, a sensitive method for the detection of NfL in serum was developed and validated. Patients with neurological diseases had higher serum NfL values than controls. In acute spinal cord injury (SCI), serum NfL levels correlated with injury severity and long-term motor outcome, and Minocycline treatment was associated with decreased NfL levels in complete SCI patients compared to placebo. Finally, I found that serum NfL levels were higher in CIS patients than in healthy controls but did not predict conversion to clinically definite MS (CDMS). Independent predictors of CDMS were instead oligoclonal bands, number of T2 lesions and age at CIS. Lower 25-OHvitamin D levels were associated with CDMS in univariate analysis, but this was attenuated in the multivariate model. In conclusion, NfL proved to be an analytically stable protein which is an important prerequisite for biomarkers. The role of NfL quantification as a surrogate measure of neuroaxonal damage is corroborated by my findings and further supports the usefulness of NfL as a putative biomarker of axonal damage in various neurological diseases.

Nearly one billion people worldwide have a neurological disease, and one out of every six adults in the United States has a mental illness. For rare and severe neurodevelopmental disorders, like Timothy syndrome (TS), exact genetic causes have been identified and studied extensively. TS is caused by a single point mutation in CACNA1C, a voltage-gated calcium channel (VGCC), which results in severe developmental defects, cardiac arrhythmia, and autism. Studies using patient derived cells are useful in identifying impaired cellular function, especially for TS and other neural activity-dependent disorders. Also, functional high-throughput screening assays that use disease-relevant cell types can lead to more targeted therapeutics that regulate cellular activity. Although these approaches are promising, cell-based assays do not consider the diversity of disease pathology or efficacy of broad-acting therapeutics in multi-cellular organisms. Therefore, we developed several whole-organism disease models using CRISPR-Cas9 and transgenes in the nematode C. elegans that harbor human VGCC mutations. We evaluated and identified behavioral, morphological, and functional phenotypes, and invented new high-throughput functional screening technologies to identify transient and potent suppressors of neural activity in these animals. We expect that these new disease models and methods will provide a pipeline for investigating activity- dependent neurological disorders in whole organisms to identify more effective therapeutics. Altogether, we aim to deepen our understanding about the brain and discover treatments for the millions of individuals that suffer from neurological disease.